Patent Application
20240043420
Provided herein are compounds and compositions for inhibition of Bcr-Abl tyrosine kinases, methods of preparing said compounds and compositions, and their use in the treatment of various cancers, such as chronic myeloid leukemia (CML).
The cytogenetic abnormality known as the Philadelphia chromosome is highly associated with the occurrence of a number of hematological malignancies, including a majority of chronic myeloid leukemias (CML) and a subset of acute lymphoblastic leukemias (Ph+ALL). The Philadelphia chromosome is a product of a translocation between the breakpoint cluster region (BCR) gene on chromosome 22 and the Abelson (ABL) tyrosine kinase gene on chromosome 9, resulting in the oncogenic fusion gene product Bcr-Abl. The resultant fusion protein is both overexpressed and harbors constitutive kinase activity that then drives the activation of a number of intracellular signaling cascades to induce the uncontrolled cell growth, division and survival associated with oncogenic transformation. Accordingly, therapeutic intervention employing inhibitors of the Bcr-Abl tyrosine kinase represents a cornerstone of the current treatment paradigm for patients with Philadelphia-positive neoplastic disorders.
Imatinib (STI-571), a small molecule Bcr-Abl tyrosine kinase inhibitor (Bcr-Abl TKI), was developed as a highly effective treatment for CML in the early 1990s and is still employed today as a first line treatment for CML. However, in more aggressive cases of CML, patients often relapse due to the emergence of resistance. The primary mechanism of this resistance derives from a variety of on-target genetic alterations that drives either aberrant overexpression of the Bcr-Abl fusion or, more commonly, introduce amino acid mutations within the Abl kinase domain that reduce imatinib's binding affinity for the active site thereby markedly reducing its inhibitory activity. These alterations can either appear stochastically and represent a sub-population within the initial tumor cell population or arise under the selective pressure of inhibitor treatment. One of the predominant on-target Bcr-Abl resistance mutations derives from point mutations that introduce an isoleucine residue for a threonine at position 315 within the Abl kinase domain (T315I) also known as the ‘gatekeeper’ position. In addition to imatinib, this mutant form of BCR-Abl is profoundly resistant to all second generation Bcr-Abl TKIs (Nilotinib, Dasatinib, Bosutinib, Radotinib). Currently, there exists only one therapeutic option for patients harboring a T315I mutation—the third line Bcr-Abl TKI, Ponatinib. While effective at treating patients with T315I CML, ponatinib suffers from poor selectivity for Bcr-Abl versus a number of other protein kinases. Accordingly, ponatinib has been reported to elicit significant dose-limiting toxicities, which then limits its ability to effectively engage the target to achieve clinical efficacy.
Besides on- or off-target resistance, intolerance to Bcr-Abl TKIs also represents a major clinical challenge. The doses of more than 50% of Ph+leukemia patients require modification due to adverse events. In fact, approximately 30% of patients are compelled to dose reduce within the first 6 months of treatment. These drug-related side effects appear early in the course of treatment and, while manageable in most cases, toxicities persist, significantly impacting the patients' quality of life, resulting in decreased compliance. Accordingly, around 40% of patients discontinue first and second generation Bcr-Abl TKIs within the first 5 years of treatment. All of the currently approved Bcr-Abl targeted therapies inhibit other tyrosine kinases, which can lead to potentially debilitating side effects. Specifically, potent inhibition of VEGFRs, PDGFRs, c-Kit and/or the c-Src family can lead to dose-limiting side effects in patients. To address these adverse effects, dose reductions, dose interruptions, and even dose discontinuations are often required during the course of therapy, however such treatment regimens ultimately result in suboptimal therapeutic benefit.
Accordingly, there remains a substantial unmet medical need for Bcr-Abl TKIs with improved selectivity to improve tolerability and enhanced potency against the wide array of resistance mechanisms in Philadelphia-positive disorders.
Provided herein are compounds and compositions that selectively inhibit Bcr-Abl tyrosine kinases and that are useful for treating disorders mediated by Bcr-Abl tyrosine kinases.
In one aspect, provided herein is a compound of formula (I),
or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, wherein:
In another aspect, provided herein is a compound of formula (I),
or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, wherein:
In some embodiments, the compound of formula (I) is a compound of formula (I-A)
In some embodiments, the compound of formula (I) is a compound of formula (I-A-i) or formula (I-A-ii)
wherein:
wherein indicates a single or double bond, and wherein the C6-C14 aryl and 5-to-10-membered heteroaryl are optionally substituted with 1-5 R6 groups;
In some embodiments, which may be combined with any of the preceding embodiments, each R1 is independently —F, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-OH, or C0-C3 alkylene-CN, wherein each pair of R4′ and R5′ of R1 taken together with the nitrogen atom to which they are attached independently form a 4-to-6-membered heterocyclic ring, wherein the heterocyclic ring optionally contains an additional 1-2 heteroatoms selected from the group consisting of N and O, and wherein each additional nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl.
In other embodiments, which may be combined with any of the preceding embodiments, R2 is phenyl,
each of which is optionally substituted with 1-5 R6 groups.
In yet other embodiments, which may be combined with any of the preceding embodiments, each R6 is independently halogen, —OR7, —NR4R5, C1-C3 alkyl, —CF3, or —CN, wherein each R4 of R6 and each R5 of R6 are independently —H or C1-C3 alkyl; and each R7 is independently —H, C1-C2 alkyl, —CD3, C1-C2 haloalkyl, or C3 cycloalkyl.
In still yet other embodiments, which may be combined with any of the preceding embodiments, each R1 is independently —F, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-OH, or C0-C3 alkylene-CN, wherein each pair of R4′ and R5′ of R1 taken together with the nitrogen atom to which they are attached independently form a 4-to-6-membered heterocyclic ring, wherein the heterocyclic ring optionally contains an additional 1-2 heteroatoms selected from the group consisting of N and O, and wherein each additional nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl;
In still other embodiments, which may be combined with any of the preceding embodiments, each R1 is independently F,
—CH2OH, —CH2CH2OH, —CN, or —CH2CN. In some embodiments, each R6 is independently —F, —Cl, —OH, —OCH3, —OCH2CH3, —OCF3, —OCF2H, —OCH2CF3, —OCD3, cyclopropyloxy, —NH2, —NHCH3, —N(CH3)2, —CH3, —CF3, or —CN.
In other embodiments of the present aspect, the compound of formula (I) is a compound of formula (I-B)
In some embodiments of the foregoing, the compound of formula (I) is a compound of formula (I-B-i) or formula (I-B-ii)
wherein:
wherein indicates a single or double bond, and wherein the C6-C14 aryl and 5-to-10-membered heteroaryl are optionally substituted with 1-5 R6 groups;
In some embodiments, which may be combined with any of the preceding embodiments, R3 is C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-CN, C1-C2 alkylene-(C3-C6 cycloalkyl), C1-C2 alkylene-O—(C1-C2 alkylene)-NR4R5, or C1-C2 alkylene-(C4-C6 heterocycloalkyl), wherein each heterocyclic nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl or C4-C6 heterocycloalkyl, wherein the R4 and R5 of R3 are independently —H or C1-C3 alkyl, and the R4′ and R5′ of R3 taken together with the nitrogen atom to which they are attached form a 4-to-6-membered heterocyclic ring, wherein the heterocyclic ring optionally contains an additional 1-2 heteroatoms selected from the group consisting of N and O, and wherein each additional nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl. In other embodiments, which may be combined with any of the preceding embodiments, R2 is phenyl,
each of which is optionally substituted with 1-5 R6 groups. In still other embodiments, which may be combined with any of the preceding embodiments, each R6 is independently halogen or —OR7; and each R7 is independently C1-C2 alkyl or C3 cycloalkyl. In yet other embodiments, which may be combined with any of the preceding embodiments, R3 is C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-CN, C1-C2 alkylene-(C3-C6 cycloalkyl), C1-C2 alkylene-O—(C1-C2 alkylene)-NR4R5, or C1-C2 alkylene-(C4-C6 heterocycloalkyl), wherein each heterocyclic nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl or C4-C6 heterocycloalkyl, wherein the R4 and R5 of R3 are independently —H or C1-C3 alkyl, and the R4′ and R5′ of R3 taken together with the nitrogen atom to which they are attached form a 4-to-6-membered heterocyclic ring, wherein the heterocyclic ring optionally contains an additional 1-2 heteroatoms selected from the group consisting of N and O, and wherein each additional nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl; R2 is phenyl,
each of which is optionally substituted with 1-5 R6 groups; each R6 is independently halogen or —OR7; and each R7 is independently C1-C2 alkyl or C3 cycloalkyl. In still yet other embodiments, which may be combined with any of the preceding embodiments, R3 is
In still further embodiments, each R6 is independently —F, —OCH3, or cyclopropyloxy. In yet other embodiments, which may be combined with any of the preceding embodiments, R1 is H.
Also provided herein is a compound which is selected from the group consisting of
or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing.
In another aspect, provided herein is a pharmaceutical composition comprising a compound of formula (I) as described herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, and one or more pharmaceutically acceptable excipients.
In yet another aspect, the present disclosure provides a method of inhibiting Bcr-Abl enzymatic activity in a cell, comprising exposing the cell with an effective amount of a compound of formula (I) as described herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, or a pharmaceutical composition comprising a compound of formula (I) as described herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing.
In still yet another aspect, provided herein is a method of treating chronic myeloid leukemia (CML), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or a mixed phenotype acute leukemia, in a human in need thereof, comprising administering to the human a compound of formula (I) as described herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, or a pharmaceutical composition comprising a compound of formula (I) as described herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing.
In some embodiments of the present aspect, the leukemia is refractory leukemia. In certain embodiments of the foregoing, the refractory leukemia is associated with a mutation in the Bcr-Abl tyrosine kinase gene resulting in specific amino acid substitutions selected from the group consisting of M244V, L248V, G250E, G250A, Q252H, Q252R, Y253F, Y253H, E255K, E255V, D276G, F311L, T315N, T315A, F317V, F317L, M343T, M351T, E355G, F359A, F359V, V379I, F382L, L387M, H396P, H396R, S417Y, E459K, F486S, and T315I. In still further embodiments of the foregoing, the refractory leukemia is associated with a mutation in the Bcr-Abl tyrosine kinase gene resulting in specific amino acid substitution T315I. In yet other embodiments, which may be combined with any preceding embodiments of the present aspect, the method further comprises administering one or more pharmaceutical agents including anti-microtubular therapies, topoisomerase inhibitors, alkylating agents, nucleotide synthesis inhibitors, DNA synthesis inhibitors, protein synthesis inhibitors, developmental signaling pathway inhibitors, pro-apoptotic agents, Abl myristoyl-pocket binding inhibitors, MEK1/2 inhibitors, AKT inhibitors, PI3K inhibitors and/or radiation.
The following description sets forth exemplary methods, parameters, and the like. It should be recognized, however, that such description is not intended as a limitation on the scope of the present disclosure but is instead provided as a description of exemplary embodiments.
As used herein, the following definitions shall apply unless otherwise indicated. Further, if any term or symbol used herein is not defined as set forth below, it shall have its ordinary meaning in the art.
The term “excipient” as used herein means an inert or inactive substance that may be used in the production of a drug or pharmaceutical, such as a tablet containing a compound of the present disclosure as an active ingredient. Various substances may be embraced by the term excipient, including without limitation any substance used as a binder, disintegrant, coating, compression/encapsulation aid, cream or lotion, lubricant, solutions for parenteral administration, materials for chewable tablets, sweetener or flavoring, suspending/gelling agent, or wet granulation agent. Binders include, e.g., carbomers, povidone, xanthan gum, etc.; coatings include, e.g., cellulose acetate phthalate, ethylcellulose, gellan gum, maltodextrin, enteric coatings, etc.; compression/encapsulation aids include, e.g., calcium carbonate, dextrose, fructose dc (dc=“directly compressible”), honey dc, lactose (anhydrate or monohydrate; optionally in combination with aspartame, cellulose, or microcrystalline cellulose), starch dc, sucrose, etc.; disintegrants include, e.g., croscarmellose sodium, gellan gum, sodium starch glycolate, etc.; creams or lotions include, e.g., maltodextrin, carrageenans, etc.; lubricants include, e.g., magnesium stearate, stearic acid, sodium stearyl fumarate, etc.; materials for chewable tablets include, e.g., dextrose, fructose dc, lactose (monohydrate, optionally in combination with aspartame or cellulose), etc.; suspending/gelling agents include, e.g., carrageenan, sodium starch glycolate, xanthan gum, etc.; sweeteners include, e.g., aspartame, dextrose, fructose dc, sorbitol, sucrose dc, etc.; and wet granulation agents include, e.g., calcium carbonate, maltodextrin, microcrystalline cellulose, etc.
The terms “individual”, “subject” and “patient” refer to mammals and includes humans and non-human mammals. Examples of patients include, but are not limited to, mice, rats, hamsters, guinea pigs, pigs, rabbits, cats, dogs, goats, sheep, cows, and humans. In some embodiments, patient refers to a human.
As used herein, the term “mammal” includes, but is not limited to, humans, mice, rats, guinea pigs, monkeys, dogs, cats, horses, cows, pigs, and sheep.
“Pharmaceutically acceptable” refers to safe and non-toxic, and suitable for in vivo or for human administration.
As used herein, the term “alkyl”, by itself or as part of another substituent, means, unless otherwise stated, a straight or branched chain hydrocarbon radical, having the number of carbon atoms designated (e.g., C1-C6 means one to six carbons). Examples of alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, t-butyl, iso-butyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. In some embodiments, the term “alkyl” may encompass C1-C6 alkyl, C2-C6 alkyl, C3-C6 alkyl, C4-C6 alkyl, C5-C6 alkyl, C1-C5 alkyl, C2-C5 alkyl, C3-C5 alkyl, C4-C5 alkyl, C1-C4 alkyl, C2-C4 alkyl, C3-C4 alkyl, C1-C3 alkyl, C2-C3 alkyl, or C1-C2 alkyl.
The term “cycloalkyl,” “carbocyclic,” or “carbocycle” refers to hydrocarbon rings having the indicated number of ring atoms (e.g., C3-C6 cycloalkyl means 3-6 carbons) and being fully saturated or having no more than one double bond between ring vertices. As used herein, “cycloalkyl,” “carbocyclic,” or “carbocycle” is also meant to refer to bicyclic, polycyclic and spirocyclic hydrocarbon rings such as, for example, bicyclo[2.2.1]heptane, pinane, bicyclo[2.2.2]octane, adamantane, norborene, spirocyclic C5-12 alkane, etc. In some embodiments, “cycloalkyl” encompasses C3-C7 cycloalkyl, C4-C7 cycloalkyl, C5-C7 cycloalkyl, C5-C7 cycloalkyl, C3-C6 cycloalkyl, C4-C6 cycloalkyl, C5-C6 cycloalkyl, C3-C5 cycloalkyl, C4-C5 cycloalkyl, or C3-C4 cycloalkyl. In addition, one ring of a polycyclic cycloalkyl group may be aromatic, provided the polycyclic cycloalkyl group is bound to the parent structure via a non-aromatic carbon. For example, a 1,2,3,4-tetrahydronaphthalen-1-yl group (wherein the moiety is bound to the parent structure via a non-aromatic carbon atom) is a cycloalkyl group, while 1,2,3,4-tetrahydronaphthalen-5-yl (wherein the moiety is bound to the parent structure via an aromatic carbon atom) is not considered a cycloalkyl group.
The term “heteroalkyl,” by itself or in combination with another term, means, unless otherwise stated, a stable straight or branched chain hydrocarbon radical, consisting of the stated number of carbon atoms and from one to three heteroatoms selected from the group consisting of O, N, Si and S, and wherein the nitrogen and sulfur atoms can optionally be oxidized and the nitrogen heteroatom can optionally be quaternized. The heteroatom(s) 0, N and S can be placed at any interior position of the heteroalkyl group. The heteroatom Si can be placed at any position of the heteroalkyl group, including the position at which the alkyl group is attached to the remainder of the molecule. A “heteroalkyl” can contain up to three units of unsaturation, and also include mono- and poly-halogenated variants, or combinations thereof. Examples include —CH2—CH2—O—CH3, —CH2—CH2—O—CF3, —CH2—CH2—NH—CH3, —CH2—CH2—N(CH3)—CH3, —CH2—S—CH2—CH3, —S(O)—CH3, —CH2—CH2—S(O)2—CH3, —CH═CH—O—CH3, —Si(CH3)3, —CH2—CH═N—OCH3, and —CH═CH═N(CH3)—CH3. Up to two heteroatoms can be consecutive, such as, for example, —CH2—NH—OCH3 and —CH2—O—Si(CH3)3.
The term “heterocycloalkyl,” “heterocyclic,” or “heterocycle” refers to a cycloalkyl radical group having the indicated number of ring atoms (e.g., 5-6 membered heterocycloalkyl) that contain from one to five heteroatoms selected from the group consisting of N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, nitrogen atom(s) are optionally quaternized, as ring atoms. Unless otherwise stated, a “heterocycloalkyl,” “heterocyclic,” or “heterocycle” ring can be a monocyclic, a bicyclic, bridged or fused ring system, spirocyclic or a polycylic ring system. Non-limiting examples of “heterocycloalkyl,” “heterocyclic,” or “heterocycle” rings include pyrrolidine, piperidine, N-methylpiperidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, piperidine, pyrimidine-2,4(1H,3H)-dione, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-5-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrhydrothiophene, quinuclidine, tropane and the like. A “heterocycloalkyl,” “heterocyclic,” or “heterocycle” group can be attached to the remainder of the molecule through one or more ring carbons or heteroatoms. In some embodiments, “heterocycloalkyl” encompasses 3- to 10-membered heterocycloalkyl, 4- to 10-membered heterocycloalkyl, 5- to 10-membered heterocycloalkyl, 6- to 10-membered heterocycloalkyl, 7- to 10-membered heterocycloalkyl, 8- to 10-membered heterocycloalkyl, 9- to 10-membered heterocycloalkyl, 3- to 9-membered heterocycloalkyl, 4- to 9-membered heterocycloalkyl, 5- to 9-membered heterocycloalkyl, 6- to 9-membered heterocycloalkyl, 7- to 9-membered heterocycloalkyl, 8- to 9-membered heterocycloalkyl, 3- to 8-membered heterocycloalkyl, 4- to 8-membered heterocycloalkyl, 5- to 8-membered heterocycloalkyl, 6- to 8-membered heterocycloalkyl, 7- to 8-membered heterocycloalkyl, 3- to 7-membered heterocycloalkyl, 4- to 7-membered heterocycloalkyl, 5- to 7-membered heterocycloalkyl, 6- to 7-membered heterocycloalkyl, 3- to 6-membered heterocycloalkyl, 4- to 6-membered heterocycloalkyl, 5- to 6-membered heterocycloalkyl, 3- to 5-membered heterocycloalkyl, 4- to 5-membered heterocycloalkyl, or 3- to 4-membered heterocycloalkyl. In other embodiments, “heterocycloalkyl” may be characterized by the number of carbon atoms in the ring, provided that the ring contains at least one heteroatom. For example, in some embodiments, “heterocycloalkyl” encompasses C3-C9 heterocycloalkyl, C3-C5 heterocycloalkyl, C3-C7 heterocycloalkyl, C3-C6 heterocycloalkyl, C3-C5 heterocycloalkyl, C3-C4 heterocycloalkyl, C4-C9 heterocycloalkyl, C4-C5 heterocycloalkyl, C4-C7 heterocycloalkyl, C4-C6 heterocycloalkyl, C4-C5 heterocycloalkyl, C5-C9 heterocycloalkyl, C3-C8 heterocycloalkyl, C5-C7 heterocycloalkyl, C5-C6 heterocycloalkyl, C6-C9 heterocycloalkyl, C6-C8 heterocycloalkyl, C6-C7 heterocycloalkyl, C7-C9 heterocycloalkyl, C7-C8 heterocycloalkyl, or C5-C9 heterocycloalkyl. It should be recognized that “heterocycloalkyl” as described by the number of ring atoms may also be described by number of carbon atoms in the ring. For example, a piperazinyl ring may be described as a C4 heterocycloalkyl ring or a 6-membered heterocycloalkyl ring; an azetidinyl or oxetanyl ring may each be described as a C3 heterocycloalkyl ring or a 4-membered heterocycloalkyl ring.
The term “alkylene” by itself or as part of another substituent means a divalent radical derived from an alkane, as exemplified by —CH2CH2CH2CH2—. Typically, an alkyl (or alkylene) group will have from 1 to 24 carbon atoms. In some embodiments, an alkyl (or alkylene) group will have 10 or fewer carbon atoms.
The term “heteroalkylene” by itself or as part of another substituent means a divalent radical, saturated or unsaturated or polyunsaturated, derived from heteroalkyl, as exemplified by —CH2—CH2—S—CH2CH2—, —CH2—S—CH2—CH2—NH—CH2—, —O—CH2—CH═CH—, —CH2—CH═C(H)CH2—O—CH2— and —S—CH2—C≡C—. For heteroalkylene groups, heteroatoms can also occupy either or both of the chain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, and the like).
The term “heterocycloalkylene” by itself or as part of another substituent means a divalent radical, saturated or unsaturated or polyunsaturated, derived from heterocycloalkyl. For heterocycloalkylene groups, heteroatoms can also occupy either or both of the chain termini.
The terms “alkoxy” and “alkylamino” are used in their conventional sense, and refer to those alkyl groups attached to the remainder of the molecule via an oxygen atom or an amino group, respectively.
The term “heterocycloalkoxy” refers to a heterocycloalkyl-O— group in which the heterocycloalkyl group is as previously described herein.
The terms “halo” or “halogen,” by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as “haloalkyl” are meant to include monohaloalkyl and polyhaloalkyl. For example, the term “C1-C4 haloalkyl” is mean to include trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, difluoromethyl, and the like.
The term “haloalkyl-OH” refers to a haloalkyl group as described above which is also substituted by one or more hydroxyl groups. The term “haloalkyl-OH” is meant to include haloalkyl substituted by one hydroxyl group, as well as haloalkyl substituted by multiple hydroxyl groups. For example, the term “haloalkyl-OH” includes —CH(F)OH, —CH2CFHCH2OH, —CH(OH)CF3, and the like.
The term “alkyl-OH” refers to an alkyl substituted by one or more hydroxyl groups. The term “alkyl-OH” is meant to include alkyl substituted by one hydroxyl group, as well as alkyl substituted by multiple hydroxyl groups. For example, the term “alkyl-OH” includes —CH2OH, —CH(OH)CH3, —CH2CH2OH, —C(CH3)2OH, and the like.
The term “alkyl-CN” refers to an alkyl substituted by one or more cyano groups. The term “alkyl-CN” is meant to include alkyl substituted by one cyano group, as well as alkyl substituted by multiple cyano groups. For example, the term “alkyl-CN” includes —CH2CN, —CH2CH2CN, —CH(CN)CH3, and the like.
The term “aryl” means, unless otherwise stated, a polyunsaturated, typically aromatic, hydrocarbon group, which can be a single ring or multiple rings (up to three rings) which are fused together. In some embodiments, “aryl” encompasses C6-C14 aryl, C5-C14 aryl, C10-C14 aryl, C12-C14 aryl, C6-C12 aryl, C5-C12 aryl, C10-C12 aryl, C6-C10 aryl, C5-C10 aryl, or C6-C8 aryl. In some embodiments, both rings of a polycyclic aryl group are aromatic (e.g., naphthyl). In other embodiments, polycyclic aryl groups may include a non-aromatic ring fused to an aromatic ring, provided the polycyclic aryl group is bound to the parent structure via an atom in the aromatic ring. Thus, in some embodiments, a 1,2,3,4-tetrahydronaphthalen-5-yl group (wherein the moiety is bound to the parent structure via an aromatic carbon atom) is considered an aryl group, while 1,2,3,4-tetrahydronaphthalen-1-yl (wherein the moiety is bound to the parent structure via a non-aromatic carbon atom) is not considered an aryl group. Similarly, in some embodiments, a 1,2,3,4-tetrahydroquinolin-8-yl group (wherein the moiety is bound to the parent structure via an aromatic carbon atom) is considered an aryl group, while 1,2,3,4-tetrahydroquinolin-1-yl group (wherein the moiety is bound to the parent structure via a non-aromatic nitrogen atom) is not considered an aryl group. However, the term “aryl” does not encompass or overlap with “heteroaryl,” as defined herein, regardless of the point of attachment (e.g., both quinolin-5-yl and quinolin-2-yl are heteroaryl groups). In some embodiments, aryl is phenyl or naphthyl. In certain embodiments, aryl is phenyl.
The term “heteroaryl” refers to aryl groups (or rings) that contain from one to five heteroatoms selected from the group consisting of N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized. A heteroaryl group can be attached to the remainder of the molecule through a carbon atom or a heteroatom as valency permits. In some embodiments, both rings of a polycyclic heteroaryl group are aromatic. In other embodiments, polycyclic heteroaryl groups may include a non-aromatic ring (e.g., cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl) fused to a heteroaryl ring, provided the polycyclic heteroaryl group is bound to the parent structure via an atom in the aromatic ring. For example, in some embodiments, a 4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl group (wherein the moiety is bound to the parent structure via an aromatic carbon atom) is considered a heteroaryl group, while 4,5,6,7-tetrahydrobenzo[d]thiazol-5-yl (wherein the moiety is bound to the parent structure via a non-aromatic carbon atom) is not considered a heteroaryl group.
Non-limiting examples of aryl groups include phenyl, naphthyl and biphenyl, while non-limiting examples of heteroaryl groups include pyridyl, pyridazinyl, pyrazinyl, pyrimindinyl, triazinyl, quinolinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalaziniyl, benzotriazinyl, purinyl, benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl, isobenzofuryl, isoindolyl, indolizinyl, benzotriazinyl, thienopyridinyl, thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridines, benzothiaxolyl, benzofuranyl, benzothienyl, indolyl, quinolyl, isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl, thiazolyl, furyl, thienyl and the like. In some embodiments, the term “heteroaryl” encompasses 5- to 10-membered heteroaryl, 6- to 10-membered heteroaryl, 7- to 10-membered heteroaryl, 8- to 10-membered heteroaryl, 9- to 10-membered heteroaryl, 5- to 9-membered heteroaryl, 6- to 9-membered heteroaryl, 7- to 9-membered heteroaryl, 8- to 9-membered heteroaryl, 5- to 8-membered heteroaryl, 6- to 8-membered heteroaryl, 7- to 8-membered heteroaryl, 5- to 7-membered heteroaryl, 6- to 7-membered heteroaryl, or 5- to 6-membered heteroaryl.
The above terms (e.g., “alkyl,” “aryl” and “heteroaryl”), in some embodiments, will include both substituted and unsubstituted forms of the indicated radical. The term “substituted” means that the specified group or moiety bears one or more substituents including, but not limited to, substituents such as alkoxy, acyl, acyloxy, alkoxycarbonyl, carbonylalkoxy, acylamino, amino, aminoacyl, aminocarbonylamino, aminocarbonyloxy, cycloalkyl, cycloalkenyl, aryl, heteroaryl, aryloxy, cyano, azido, halo, hydroxyl, nitro, carboxyl, thiol, thioalkyl, alkyl, alkenyl, alkynyl, heterocycloalkyl, heterocycloalkenyl, aralkyl, aminosulfonyl, sulfonylamino, sulfonyl, oxo and the like. The term “unsubstituted” means that the specified group bears no substituents. Where the term “substituted” is used to describe a structural system, the substitution is meant to occur at any valency-allowed position on the system. When a group or moiety bears more than one substituent, it is understood that the substituents may be the same or different from one another. In some embodiments, a substituted group or moiety bears from one to five substituents. In some embodiments, a substituted group or moiety bears one substituent. In some embodiments, a substituted group or moiety bears two substituents. In some embodiments, a substituted group or moiety bears three substituents. In some embodiments, a substituted group or moiety bears four substituents. In some embodiments, a substituted group or moiety bears five substituents.
By “optional” or “optionally” is meant that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances in which it does not. For example, “optionally substituted alkyl” encompasses both “alkyl” and “substituted alkyl” as defined herein. It will be understood by those skilled in the art, with respect to any group containing one or more substituents, that such groups are not intended to introduce any substitution or substitution patterns that are sterically impractical, synthetically non-feasible, and/or inherently unstable. It will also be understood that where a group or moiety is optionally substituted, the disclosure includes both embodiments in which the group or moiety is substituted and embodiments in which the group or moiety is unsubstituted.
As used herein, the term “heteroatom” is meant to include oxygen (O), nitrogen (N), sulfur (S) and silicon (Si).
As used herein, the term “chiral” refers to molecules which have the property of non-superimposability of the mirror image partner, while the term “achiral” refers to molecules which are superimposable on their mirror image partner.
As used herein, the term “stereoisomers” refers to compounds which have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space.
As used herein, a wavy line “” that intersects a bond in a chemical structure indicates the point of attachment of the atom to which the wavy bond is connected in the chemical structure to the remainder of a molecule, or to the remainder of a fragment of a molecule.
As used herein, the representation of a group (e.g., Xa) in parenthesis followed by a subscript integer range (e.g., (Xa)0-1) means that the group can have the number of occurrences as designated by the integer range. For example, (Xa)0-1 means the group Xa can be absent or can occur one time.
“Diastereomer” refers to a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities. Mixtures of diastereomers can separate under high resolution analytical procedures such as electrophoresis and chromatography.
“Enantiomers” refer to two stereoisomers of a compound which are non-superimposable mirror images of one another.
Stereochemical definitions and conventions used herein generally follow S. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., “Stereochemistry of Organic Compounds”, John Wiley & Sons, Inc., New York, 1994. The compounds of the present disclosure can contain asymmetric or chiral centers, and therefore exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of the present disclosure, including but not limited to, diastereomers, enantiomers and atropisomers, as well as mixtures thereof such as racemic mixtures, form part of the present disclosure. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L, or R and S, are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes d and 1 or (+) and (−) are employed to designate the sign of rotation of plane-polarized light by the compound, with (−) or 1 meaning that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of one another. A specific stereoisomer can also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture or a racemate, which can occur where there has been no stereoselection or stereospecificity in a chemical reaction or process. The terms “racemic mixture” and “racemate” refer to an equimolar mixture of two enantiomeric species, devoid of optical activity.
As used herein, the term “tautomer” or “tautomeric form” refers to structural isomers of different energies which are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol and imine-enamine isomerizations. Valence tautomers include interconversions by reorganization of some of the bonding electrons.
As used herein, the term “solvate” refers to an association or complex of one or more solvent molecules and a compound of the present disclosure. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine. The term “hydrate” refers to the complex where the solvent molecule is water. Certain compounds of the present disclosure can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present disclosure.
The term “co-crystal” as used herein refers to a solid that is a crystalline single phase material composed of two or more different molecular or ionic compounds generally in a stoichiometric ratio which are neither solvates nor simple salts. A co-crystal consists of two or more components that form a unique crystalline structure having unique properties. Co-crystals are typically characterized by a crystalline structure, which is generally held together by freely reversible, non-covalent interactions. As used herein, a co-crystal refers to a compound of the present disclosure and at least one other component in a defined stoichiometric ratio that form a crystalline structure.
As used herein, the term “protecting group” refers to a substituent that is commonly employed to block or protect a particular functional group on a compound. For example, an “amino-protecting group” is a substituent attached to an amino group that blocks or protects the amino functionality in the compound. Suitable amino-protecting groups include acetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ) and 9-fluorenylmethylenoxycarbonyl (Fmoc). Similarly, a “hydroxy-protecting group” refers to a substituent of a hydroxy group that blocks or protects the hydroxy functionality. Suitable protecting groups include acetyl and silyl. A “carboxy-protecting group” refers to a substituent of the carboxy group that blocks or protects the carboxy functionality. Common carboxy-protecting groups include phenylsulfonylethyl, cyanoethyl, 2-(trimethylsilyl)ethyl, 2-(trimethylsilyl)ethoxymethyl, 2-(p-toluenesulfonyl)ethyl, 2-(p-nitrophenylsulfenyl)ethyl, 2-(diphenylphosphino)-ethyl, nitroethyl and the like. For a general description of protecting groups and their use, see P. G. M. Wuts and T. W. Greene, Greene's Protective Groups in Organic Synthesis 4th edition, Wiley-Interscience, New York, 2006.
As used herein, the term “pharmaceutically acceptable salts” is meant to include salts of the active compounds which are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein. When compounds of the present disclosure contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of salts derived from pharmaceutically-acceptable inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, zinc and the like. Salts derived from pharmaceutically-acceptable organic bases include salts of primary, secondary and tertiary amines, including substituted amines, cyclic amines, naturally-occurring amines and the like, such as arginine, betaine, caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like. When compounds of the present disclosure contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, malonic, benzoic, succinic, suberic, fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, for example, Berge, S. M., et al., “Pharmaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present disclosure contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.
The neutral forms of the compounds can be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but otherwise the salts are equivalent to the parent form of the compound for the purposes of the present disclosure.
Certain compounds of the present disclosure possess asymmetric carbon atoms (optical centers) or double bonds; the racemates, diastereomers, geometric isomers, regioisomers and individual isomers (e.g., separate enantiomers) are all intended to be encompassed within the scope of the present disclosure.
The compounds of the present disclosure can also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the present disclosure also embraces isotopically-labeled variants of the present disclosure which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having the atomic mass or mass number different from the predominant atomic mass or mass number usually found in nature for the atom. All isotopes of any particular atom or element as specified are contemplated within the scope of the compounds of the present disclosure and include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, chlorine and iodine, such as 2H (“D”), 3H, 11C, 13C, 14C, 13N, 15N, 15O, 17O, 18O, 32P, 33P, 35S, 18F, 36Cl, 123I and 125I. Certain isotopically labeled compounds of the present disclosure (e.g., those labeled with 3H or 14C) are useful in compound and/or substrate tissue distribution assays. Tritiated (3H) and carbon-14 (14C) isotopes are useful for their ease of preparation and detectability. Further substitution with heavier isotopes such as deuterium (i.e., 2H) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and hence may be preferred in some circumstances. Positron emitting isotopes such as 15O, 13N, 11C, and 18F are useful for positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds of the present disclosure can generally be prepared by following procedures analogous to those disclosed in the Schemes and/or in the Examples herein below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.
“Treating” or “treatment” of a disease in a patient refers to inhibiting the disease or arresting its development; or ameliorating or causing regression of the disease. As used herein, “treatment” or “treating” is an approach for obtaining beneficial or desired results including clinical results. For purposes of this disclosure, beneficial or desired results include, but are not limited to, one or more of the following: decreasing one more symptoms resulting from the disease or disorder, diminishing the extent of the disease or disorder, stabilizing the disease or disorder (e.g., preventing or delaying the worsening of the disease or disorder), delaying the occurrence or recurrence of the disease or disorder, delay or slowing the progression of the disease or disorder, ameliorating the disease or disorder state, providing a remission (whether partial or total) of the disease or disorder, decreasing the dose of one or more other medications required to treat the disease or disorder, enhancing the effect of another medication used to treat the disease or disorder, delaying the progression of the disease or disorder, increasing the quality of life, and/or prolonging survival of a patient. Also encompassed by “treatment” is a reduction of pathological consequence of the disease or disorder. The methods of the present disclosure contemplate any one or more of these aspects of treatment.
“Preventing”, “prevention”, or “prophylaxis” of a disease in a patient refers to preventing the disease from occurring in a patient that is predisposed or does not yet display symptoms of the disease.
The phrase “therapeutically effective amount” means an amount of a compound of the present disclosure that (i) treats or prevents the particular disease, condition, or disorder, (ii) attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition, or disorder described herein.
The terms “cancer” and “cancerous” refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth.
It is appreciated that certain features of the present disclosure, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination. All combinations of the embodiments pertaining to the chemical groups represented by the variables are specifically embraced by the present invention and are disclosed herein just as if each and every combination was individually and explicitly disclosed, to the extent that such combinations embrace compounds that are stable compounds (i.e., compounds that can be isolated, characterized, and tested for biological activity). In addition, all subcombinations of the chemical groups listed in the embodiments describing such variables are also specifically embraced by the present invention and are disclosed herein just as if each and every such sub-combination of chemical groups was individually and explicitly disclosed herein.
In one aspect, provided herein is a compound of formula (I),
or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, wherein:
In one aspect, provided herein is a compound of formula (I)
or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, wherein:
In some embodiments of the present aspect, R0 is
In some embodiments wherein R0 is
m is an integer 0, 1, 2 or 3. In some embodiments, m is 0. In other embodiments, m is 1. In yet other embodiments, m is 2. In still yet other embodiments, m is 3. In other embodiments of the present aspect, R0 is
In some embodiments, the compound of formula (I) is a compound of formula (I-A) or formula (I-B)
In some embodiments of the present aspect, X is N or CR8. In some embodiments, X is N. In some embodiments, X is CR8.
In some embodiments, each R1 is independently -D, —F, C1-C3 alkyl, C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, —O—C1-C3 alkylene-NR4R5, —O—C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-OH, C0-C3 alkylene-CN, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4′R5′, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-OH, C1-C2 alkylene-(4- to 8-membered heterocycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C2 alkylene-(4- to 8-membered heterocycloalkylene)-(C0-C2 alkylene)-NR4′R5, C1-C2 alkylene-(C3-C7 heterocycloalkylene)-(C0-C2 alkylene)-NR4R5 or C1-C2 alkylene-(C3-C7 heterocycloalkylene)-(C0-C2 alkylene)-NR4′R5′, wherein the alkyl, alkylene, cycloalkylene, and heterocycloalkylene moieties in R1 are optionally substituted with 1-3 fluorine atoms and/or 1-6 deuterium atoms, and wherein each heterocyclic nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl, C3-C6 cycloalkyl, C2-C3 haloalkyl, C2-C3 alkylene-CN, or C2-C3 heteroalkyl.
In some embodiments, each R1 is independently -D, —F, C1-C3 alkyl, C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-OH, C0-C3 alkylene-CN, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4′R5′, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-OH, C1-C2 alkylene-(C4-C6 heterocycloalkylene)-(C0-C2 alkylene)-NR4R5 or C1-C2 alkylene-(C4-C6 heterocycloalkylene)-(C0-C2 alkylene)-NR4′R5′, wherein the alkyl, alkylene, cycloalkylene, and heterocycloalkylene moieties in R1 are optionally substituted with 1-3 fluorine atoms and/or 1-6 deuterium atoms, and wherein each heterocyclic nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl, C3-C6 cycloalkyl, C2-C3 haloalkyl, C2-C3 alkylene-CN, or C2-C3 heteroalkyl. In some embodiments, each R1 is independently —F, C1-C3 alkyl, C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-OH, C0-C3 alkylene-CN, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4′R5′, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-OH, C1-C2 alkylene-(C4-C6 heterocycloalkylene)-(C0-C2 alkylene)-NR4R5 or C1-C2 alkylene-(C4-C6 heterocycloalkylene)-(C0-C2 alkylene)-NR4′R5′, wherein the alkyl, alkylene, cycloalkylene, and heterocycloalkylene moieties in R1 are optionally substituted with 1-3 fluorine atoms and/or 1-6 deuterium atoms, and wherein each heterocyclic nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl, C3-C6 cycloalkyl, C2-C3 haloalkyl, C2-C3 alkylene-CN, or C2-C3 heteroalkyl. In some embodiments, each R1 is independently -D, —F, or C1-C3 alkyl. In some embodiments, each R1 is independently C1-C3 alkyl, C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, —O—C1-C3 alkylene-NR4R5, —O—C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-OH, or C0-C3 alkylene-CN. In some embodiments, each R1 is independently C1-C3 alkyl, C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-OH, or C0-C3 alkylene-CN. In some embodiments, each R1 is independently C1-C3 alkyl, C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, —O—C1-C3 alkylene-NR4R5, or —O—C1-C3 alkylene-NR4′R5′. In some embodiments, each R1 is independently C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4′R5′, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-OH, C1-C2 alkylene-(4- to 8-membered heterocycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C2 alkylene-(4- to 8-membered heterocycloalkylene)-(C0-C2 alkylene)-NR4′R5, C1-C2 alkylene-(C3-C7 heterocycloalkylene)-(C0-C2 alkylene)-NR4R5 or C1-C2 alkylene-(C3-C7 heterocycloalkylene)-(C0-C2 alkylene)-NR4′R5′. In some embodiments, each R1 is independently C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4′R5′, C1-C2 alkylene-(C4-C6 heterocycloalkylene)-(C0-C2 alkylene)-NR4R5 or C1-C2 alkylene-(C4-C6 heterocycloalkylene)-(C0-C2 alkylene)-NR4′R5′. In some embodiments, each R1 is independently C1-C3 alkylene-OH, C0-C3 alkylene-CN, or C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-OH. In some embodiments, each R1 is independently —F, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-OH, or C0-C3 alkylene-CN, wherein each pair of R4′ and R5′ of R1 taken together with the nitrogen atom to which they are attached independently form a 3-to-7-membered heterocyclic ring, wherein the heterocyclic ring optionally contains an additional 1-2 heteroatoms selected from the group consisting of N, O, and S, and wherein each additional nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl, C3-C6 cycloalkyl, C2-C3 haloalkyl, C2-C3 alkylene-CN, or C2-C3 heteroalkyl. In some embodiments, each R1 is independently —F, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-OH, or C0-C3 alkylene-CN, wherein each pair of R4′ and R5′ of R1 taken together with the nitrogen atom to which they are attached independently form a 4-to-6-membered heterocyclic ring, wherein the heterocyclic ring optionally contains an additional 1-2 heteroatoms selected from the group consisting of N and O, and wherein each additional nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl. In some embodiments, each R1 is independently C1-C3 alkylene-NR4′R5′. In certain embodiments, each R1 is independently optionally substituted —C1-C2 alkylene-N-morpholinyl or optionally substituted —C1-C2 alkylene-N-piperazinyl. In some embodiments, each R1 is independently optionally substituted
optionally substituted
optionally substituted
optionally substituted
optionally substituted
or optionally substituted
In certain embodiments, each R1 is independently optionally substituted
optionally substituted
optionally substituted
or optionally substituted
In some embodiments, each R1 is independently
In certain embodiments, each R1 is independently
In certain other embodiments, each R1 is independently,
In still other embodiments, each R1 is independently
In some embodiments, each R1 is independently C1-C3 alkylene-OH. In certain other embodiments, each R1 is independently —C1-C2 alkylene-OH. In certain embodiments, each R1 is independently —CH2OH, —CH2CH2OH, —CH(OH)CH3, —CH2CH2CH2OH, —CH2CH(OH)CH3, or —CH(CN)CH2CH3. In certain other embodiments, each R1 is independently —CH2OH or —CH2CH2OH. In some embodiments, each R1 is independently C0-C3 alkylene-CN. In certain embodiments, each R1 is independently —CN, —CH2CN, —CH2CH2CN, —CH(CN)CH3, —CH2CH2CH2CN, —CH2CH(CN)CH3, —CH(CN)CH2CH3, or —CH(CH2CN)CH3. In some embodiments, each R1 is independently —F,
—CH2OH, —CH2CH2OH, —CH2—NHCH3, —CH2NHCD3, —CH2—N(CH3)2, —CH2N(CD3)2, —CH2CH2—N(CH3)2, —CH2CH2N(CD3)2, —OCH2CH2N(CH3)2, —CN, or —CH2CN. In some embodiments, each R1 is independently —F,
—CH2OH, —CH2CH2OH, —CN, or —CH2CN.
In other embodiments, two R1 are taken together with the carbon atom or carbon atoms to which they are attached to form a 3- to 7-membered heterocyclic ring, wherein the heterocyclic ring contains nitrogen atom and wherein the nitrogen atom is optionally substituted with C1-C3 alkyl.
For example, in some embodiments, two R1 are present on adjacent carbon atoms and together with the carbon atoms to which they are attached form a 3- to 7-membered heterocyclic ring, such as a pyrrolidinyl ring, wherein the heterocyclic ring contains nitrogen atom and wherein the nitrogen atom is optionally substituted with C1-C3 alkyl. In other embodiments, two R1 are present on the same carbon atom and together with the carbon atoms to which they are attached form a 3- to 7-membered heterocyclic ring, such as a azetidinyl ring, wherein the heterocyclic ring contains nitrogen atom and wherein the nitrogen atom is optionally substituted with C1-C3 alkyl.
In some embodiments, R2 is C6-C14 aryl or 5-to-10-membered heteroaryl, wherein the C6-C14 aryl and 5-to-10-membered heteroaryl are optionally substituted with 1-5 R6 groups. In some embodiments, R2 is a C6-C14 aryl, wherein said C6-C14 aryl is optionally substituted with 1-5 R6 groups. In some embodiments, R2 is phenyl, wherein said phenyl is optionally substituted with 1-5 R6 groups. In some embodiments, R2 is 5- to 10-membered heteroaryl, wherein said 5-to-10-membered heteroaryl is optionally substituted with 1-5 R6 groups. In some embodiments, R2 is a 5-to-10-membered heteroaryl selected from the group consisting of
wherein indicates a single or double bond, and wherein the 5-to-10-membered heteroaryl is optionally substituted with 1-5 R6 groups. In some embodiments, R2 is
each of which is optionally substituted with 1-5 R6 groups. In some embodiments, R2 is a 5-to-10-membered heteroaryl selected from the group consisting of
wherein indicates a single or double bond, and wherein the 5-to-10-membered heteroaryl is optionally substituted with 1-5 R6 groups. In some embodiments, R2 is
each of which is optionally substituted with 1-5 R6 groups. In some embodiments, R2 is phenyl,
each of which is optionally substituted with 1-5 R6 groups.
In some embodiments, R3 is —H, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkylene-NR4R5, C1-C6 alkylene-NR4′R5′, C1-C6 alkylene-OH, C1-C3 alkylene-CN, C1-C3 alkylene-(C3-C6 cycloalkyl), C1-C3 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C3 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4′R5′, C1-C3 alkylene-O—(C1-C2 alkylene)-NR4R5, C1-C3 alkylene-O—(C1-C3 alkylene)-NR4′R5′, C1-C3 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-OH, C1-C3 alkylene-(4- to 8-membered heterocycloalkyl), C1-C3 alkylene-(C3-C7 heterocycloalkyl), C1-C3 alkylene-(4- to 8-membered heterocycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C3 alkylene-(4- to 8-membered heterocycloalkylene)-(C0-C2 alkylene)-NR4′R5′, (4- to 8-membered heterocycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C3 alkylene-(C3-C7 heterocycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C3 alkylene-(C3-C7 heterocycloalkylene)-(C0-C2 alkylene)-NR4′R5′, or (C3-C7 heterocycloalkylene)-(C0-C2 alkylene)-NR4R5, wherein the alkyl, alkylene, cycloalkyl, cycloalkylene, heterocycloalkyl, and heterocycloalkylene moieties in R3 are optionally substituted with 1-3 fluorine atoms, 1-3 CN groups and/or 1-6 deuterium atoms and wherein each heterocyclic nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl, C3-C6 cycloalkyl, C2-C3 haloalkyl, C2-C3 alkylene-CN, C2-C3 heteroalkyl, 4- to 8-membered heterocycloalkyl, or C3-C7 heterocycloalkyl.
In some embodiments, R3 is —H, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkylene-NR4R5, C1-C6 alkylene-NR4′R5′, C1-C6 alkylene-OH, C1-C3 alkylene-CN, C1-C3 alkylene-(C3-C6 cycloalkyl), C1-C3 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C3 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4′R5′, C1-C3 alkylene-O—(C1-C2 alkylene)-NR4R5, C1-C3 alkylene-O—(C1-C3 alkylene)-NR4′R5′, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-OH, C1-C3 alkylene-(C4-C6 heterocycloalkyl), C1-C3 alkylene-(C4-C6 heterocycloalkylene)-(C0-C2 alkylene)-NR4R5 or C1-C3 alkylene-(C4-C6 heterocycloalkylene)-(C0-C2 alkylene)-NR4′R5′, wherein the alkyl, alkylene, cycloalkyl, cycloalkylene, heterocycloalkyl, and heterocycloalkylene moieties in R3 are optionally substituted with 1-3 fluorine atoms and/or 1-6 deuterium atoms and wherein each heterocyclic nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl, C3-C6 cycloalkyl, C2-C3 haloalkyl, C2-C3 alkylene-CN, C2-C3 heteroalkyl, or C4-C6 heterocycloalkyl.
In some embodiments, R3 is C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-OH, C1-C3 alkylene-CN, C1-C2 alkylene-(C3-C6 cycloalkyl), C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-NR4′R5′, C1-C2 alkylene-O—(C1-C2 alkylene)-NR4R5, C1-C2 alkylene-(C3-C6 cycloalkylene)-(C0-C2 alkylene)-OH, C1-C2 alkylene-(C4-C6 heterocycloalkyl), C1-C2 alkylene-(C4-C6 heterocycloalkylene)-(C0-C2 alkylene)-NR4R5, or C1-C2 alkylene-(C4-C6 heterocycloalkylene)-(C0-C2 alkylene)-NR4′R5′, wherein the alkyl, alkylene, cycloalkyl, cycloalkylene, heterocycloalkyl, and heterocycloalkylene moieties in R3 are optionally substituted with 1-3 fluorine atoms and/or 1-6 deuterium atoms and wherein each heterocyclic nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl, C3-C6 cycloalkyl, C2-C3 haloalkyl, C2-C3 alkylene-CN, C2-C3 heteroalkyl, or C4-C6 heterocycloalkyl. In some embodiments, R3 is C1-C3 alkylene-(4- to 8-membered heterocycloalkyl), C1-C3 alkylene-(C3-C7 heterocycloalkyl), C1-C3 alkylene-(4- to 8-membered heterocycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C3 alkylene-(4- to 8-membered heterocycloalkylene)-(C0-C2 alkylene)-NR4′R5′, (4- to 8-membered heterocycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C3 alkylene-(C3-C7 heterocycloalkylene)-(C0-C2 alkylene)-NR4R5, C1-C3 alkylene-(C3-C7 heterocycloalkylene)-(C0-C2 alkylene)-NR4′R5′, or (C3-C7 heterocycloalkylene)-(C0-C2 alkylene)-NR4R5 In some embodiments, R3 is C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-CN, C1-C2 alkylene-(C3-C6 cycloalkyl), C1-C2 alkylene-O—(C1-C2 alkylene)-NR4R5, C1-C3 alkylene-(4- to 8-membered heterocycloalkyl), or C1-C3 alkylene-(C3-C7 heterocycloalkyl), wherein each heterocyclic nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl or C4-C6 heterocycloalkyl, wherein the R4 and R5 of R3 taken together with the nitrogen atom to which they are attached independently form a 3-to-7-membered heterocyclic ring, wherein the heterocyclic ring optionally contains an additional 1-2 heteroatoms selected from the group consisting of N, O, and S, and wherein each additional nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl, C3-C6 cycloalkyl, C2-C3 haloalkyl, C2-C3 alkylene-CN, or C2-C3 heteroalkyl. In some embodiments, R3 is C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-CN, C1-C2 alkylene-(C3-C6 cycloalkyl), C1-C2 alkylene-O—(C1-C2 alkylene)-NR4R5, or C1-C2 alkylene-(C4-C6 heterocycloalkyl), wherein each heterocyclic nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl or C4-C6 heterocycloalkyl, wherein the R4 and R5 of R3 are independently —H or C1-C3 alkyl, and the R4′ and R5′ of R3 taken together with the nitrogen atom to which they are attached form a 4-to-6-membered heterocyclic ring, wherein the heterocyclic ring optionally contains an additional 1-2 heteroatoms selected from the group consisting of N and O, and wherein each additional nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl. In some embodiments, each R1 is independently optionally substituted
optionally substituted
optionally substituted
optionally substituted
optionally substituted
or optionally substituted
In certain embodiments, each R1 is independently optionally substituted
optionally substituted
optionally substituted
or optionally substituted
In some embodiments, each R1 is independently
In certain embodiments, each R1 is independently
In certain embodiments, each R1 is independently
In some embodiments R3 is,
In some embodiments, R3 is
In some embodiments, each R4 is independently —H, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkylene-CN, or C1-C6 heteroalkyl, wherein said C1-C3 alkyl is optionally substituted with 1-6 deuterium atoms. In some embodiments, each R4 is independently —H, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkylene-CN, or C1-C6 heteroalkyl. In some embodiments, each R4 is independently —H, C1-C3 alkyl, C3-C6 cycloalkyl, C2-C3 haloalkyl, C2-C3 alkylene-CN, or C2-C3 heteroalkyl. In some embodiments, each R4 is independently —H. In some embodiments, each R4 is independently C1-C3 alkyl. In certain embodiments, each R4 is independently —CH3, —CH2CH3, —CH2CH2CH3, or —CH(CH3)2. In some embodiments, each R4 is independently C3-C6 cycloalkyl. In certain embodiments, R4 is independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In some embodiments, each R4 is independently C2-C3 haloalkyl. In certain embodiment, each R4 is independently C2-C3 haloalkyl, wherein the each halogen atom of each C2-C3 haloalkyl is independently —F, —C1, or —Br. In some embodiments, each R4 is independently C2-C3 alkylene-CN. In certain embodiments, each R4 is independently —CH2CH2CN, —CH(CN)CH3, —CH2CH2CH2CN, —CH2CH(CN)CH3, —CH(CN)CH2CH3, or —CH(CH2CN)CH3. In some embodiments, each R4 is independently C2-C3 heteroalkyl. In certain embodiments, each R4 is independently —CH2CH2OH, —CH(OH)CH3, —CH2CH2CH2OH, —CH2CH(OH)CH3, or —CH(OH)CH2CH3, —CH2OCH3, —CH2OCH2CH3, —CH2CH2OCH3, —CH(OCH3)CH3, —CH(CH2OH)CH3, —CH2CH2 NH2, —CH(NH2)CH3, —CH2CH2CH2 NH2, —CH2CH(NH2)CH3, or —CH(NH2)CH2CH3, —CH2NHCH3, —CH2NHCH2CH3, —CH2CH2NHCH3, —CH(NHCH3)CH3, —CH(CH2NH2CH3, —CH2CH2SH, —CH(SH)CH3, —CH2CH2CH2SH, —CH2CH(SH)CH3, or —CH(SH)CH2CH3, —CH2SCH3, —CH2SCH2CH3, —CH2CH2SCH3, —CH(SCH3)CH3, or —CH(CH2SH)CH3.
In some embodiments, each R5 is independently —H, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkylene-CN, or C1-C6 heteroalkyl, wherein said C1-C3 alkyl is optionally substituted with 1-6 deuterium atoms. In some embodiments, each R5 is independently —H, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkylene-CN, or C1-C6 heteroalkyl. In some embodiments, each R5 is independently —H, C1-C3 alkyl, C3-C6 cycloalkyl, C2-C3 haloalkyl, C2-C3 alkylene-CN, or C2-C3 heteroalkyl. In some embodiments, each R5 is independently —H. In some embodiments, each R5 is independently C1-C3 alkyl. In certain embodiments, each R5 is independently —CH3, —CH2CH3, —CH2CH2CH3, or —CH(CH3)2. In some embodiments, each R5 is independently C3-C6 cycloalkyl. In certain embodiments, R5 is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In some embodiments, each R5 is independently C2-C3 haloalkyl. In certain embodiment, each R5 is independently C2-C3 haloalkyl, wherein the each halogen atom of each C2-C3 haloalkyl is independently —F, —C1, or —Br. In some embodiments, each R5 is independently C2-C3 alkylene-CN. In certain embodiments, each R5 is independently —CH2CH2CN, —CH(CN)CH3, —CH2CH2CH2CN, —CH2CH(CN)CH3, —CH(CN)CH2CH3, or —CH(CH2CN)CH3. In some embodiments, each R5 is independently C2-C3 heteroalkyl. In certain embodiments, each R5 is independently —CH2CH2OH, —CH(OH)CH3, —CH2CH2CH2OH, —CH2CH(OH)CH3, or —CH(OH)CH2CH3, —CH2OCH3, —CH2OCH2CH3, —CH2CH2OCH3, —CH(OCH3)CH3, —CH(CH2OH)CH3, —CH2CH2 NH2, —CH(NH2)CH3, —CH2CH2CH2 NH2, —CH2CH(NH2)CH3, or —CH(NH2)CH2CH3, —CH2NHCH3, —CH2NHCH2CH3, —CH2CH2NHCH3, —CH(NHCH3)CH3, —CH(CH2NH2CH3, —CH2CH2SH, —CH(SH)CH3, —CH2CH2CH2SH, —CH2CH(SH)CH3, or —CH(SH)CH2CH3, —CH2SCH3, —CH2SCH2CH3, —CH2CH2SCH3, —CH(SCH3)CH3, or —CH(CH2SH)CH3.
In some embodiments, each pair of R4′ and R5′ taken together with the nitrogen atom to which they are attached independently form a 3-to-7-membered heterocyclic ring, wherein the heterocyclic ring optionally contains an additional 1-2 heteroatoms selected from the group consisting of N, O, and S, and wherein each additional nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl, C3-C6 cycloalkyl, C2-C3 haloalkyl, C2-C3 alkylene-CN, or C2-C3 heteroalkyl. In some embodiments, each pair of R4′ and R5′ taken together with the nitrogen atom to which they are attached independently form a 4-to-6-membered heterocyclic ring, wherein the heterocyclic ring optionally contains an additional 1-2 heteroatoms selected from the group consisting of N, O, and S, and wherein each additional nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl, C3-C6 cycloalkyl, C2-C3 haloalkyl, C2-C3 alkylene-CN, or C2-C3 heteroalkyl. In some embodiments, each pair of R4′ and R5′ taken together with the nitrogen atom to which they are attached independently form a 4-to-6-membered heterocyclic ring, wherein the heterocyclic ring optionally contains an additional 1-2 heteroatoms selected from the group consisting of N and O, and wherein each additional nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl, C3-C6 cycloalkyl, C2-C3 haloalkyl, C2-C3 alkylene-CN, or C2-C3 heteroalkyl.
In some embodiments, each R6 is independently halogen, —OR7, —NR4R5, —NR4′R5′, C1-C3 alkyl, C1-C3 haloalkyl, C3-C6 cycloalkyl, —CN, S(O)nC1-C3 alkyl, or S(O)nC3-C6 cycloalkyl, wherein n is an integer from 0 to 2. In some embodiments, each R6 is independently halogen, —OR7, —NR4R5, C1-C3 alkyl, —CF2H, —CF3, C3-C6 cycloalkyl, or —CN. In some embodiments, each R6 is independently halogen, —OR7, —NR4R5, C1-C3 alkyl, —CF3, or —CN, wherein each R4 of R6 and each R5 of R6 are independently —H or C1-C3 alkyl. In some embodiments, each R6 is independently —F, —Cl, —OH, —OCH3, —OCH2CH3, —OCF3, —OCF2H, —OCH2CF3, —OCD3, cyclopropyloxy, cyclobutoxy, —O—CH2-cyclopropyl, —NH2, —NHCH3, —N(CH3)2, —CH3, —CF3, or —CN. In some embodiments, each R6 is independently —F, —Cl, —OH, —OCH3, —OCH2CH3, —OCF3, —OCF2H, —OCH2CF3, —OCD3, cyclopropyloxy, —NH2, —NHCH3, —N(CH3)2, —CH3, —CF3, or —CN. In some embodiments, each R6 is independently halogen or —OR7. In some embodiments, each R6 is independently —F, —OCH3, or cyclopropyloxy.
In some embodiments, each R7 is independently —H, C1-C3 alkyl, C1-C3 haloalkyl, C3-C6 cycloalkyl, or C1-C3-alkylene-C3-C6 cycloalkyl, wherein said C1-C3 alkyl is optionally substituted with 1-6 deuterium atoms. In some embodiments, each R7 is independently —H, C1-C3 alkyl, C1-C3 haloalkyl, or C3-C6 cycloalkyl, wherein said C1-C3 alkyl is optionally substituted with 1-6 deuterium atoms. In some embodiments, each R7 is independently —H, C1-C3 alkyl, —CD3, C1-C2 haloalkyl, or C3-C6 cycloalkyl. In some embodiments, each R7 is independently —H, C1-C3 alkyl, —CD3, —CF2H, —CF3, or C3-C6 cycloalkyl. In some embodiments, each R7 is independently —H, C1-C2 alkyl, —CD3, C1-C2 haloalkyl, or C3 cycloalkyl. In some embodiments, each R7 is independently C1-C2 alkyl or C3 cycloalkyl.
In some embodiments, R8 is —H, —F, or C1-C3 alkyl. In some embodiments, R8 is —H, —F, or —CH3. In some embodiments, R8 is —H or —F. In some embodiments, R8 is —H or C1-C3 alkyl. In some embodiments, R8 is —F or C1-C3 alkyl. In some embodiments, R8 is —H, —F, —CH3, —CH2CH3, —CH2CH2CH3, or —CH(CH3)2. In some embodiments, R8 is —H. In some embodiments, R8 is —F. In some embodiments, R8 is C1-C3 alkyl. In some embodiments, —CH3, —CH2CH3, —CH2CH2CH3, or —CH(CH3)2. In some embodiments, R1 is —CH3.
In some embodiments, the compound of formula (I) is a compound of formula (I-A), or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing
In some embodiments, the compound of formula (I) or formula (I-A) is a compound of formula (I-A-i) or formula (I-A-ii)
wherein
wherein indicates a single or double bond, and wherein the C6-C14 aryl and 5-to-10-membered heteroaryl are optionally substituted with 1-5 R6 groups;
In some embodiments of the compound of formula (I), formula (I-A), formula (I-A-i),
In some embodiments, R2 is phenyl,
each of which is optionally substituted with 1-5 R6 groups.
In some embodiments, each R6 is independently halogen, —OR7, —NR4R5, C1-C3 alkyl, —CF3, or —CN, wherein each R4 of R6 and each R5 of R6 are independently —H or C1-C3 alkyl; and each R7 is independently —H, C1-C2 alkyl, —CD3, C1-C2 haloalkyl, or C3 cycloalkyl.
In some embodiments, each R1 is independently —F, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-OH, or C0-C3 alkylene-CN, wherein each pair of R4′ and R5′ of R1 taken together with the nitrogen atom to which they are attached independently form a 4-to-6-membered heterocyclic ring, wherein the heterocyclic ring optionally contains an additional 1-2 heteroatoms selected from the group consisting of N and O, and wherein each additional nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl;
In some embodiments, each R1 is independently F, N—CH2OH, —CH2CH2OH, —CN, or —CH2CN.
In some embodiments of the compound of formula (I), formula (I-A), formula (I-A-i), each R6 is independently —F, —Cl, —OH, —OCH3, —OCH2CH3, —OCF3, —OCF2H, —OCH2CF3, OCD3, cyclopropyloxy, —NH2, —NHCH3, —N(CH3)2, —CH3, —CF3, or —CN.
In some embodiments, the compound of formula (I) is a compound of formula (I-B), or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing
In some embodiments, wherein the compound of formula (I) or formula (I-B) is a compound of formula (I-B-i) or formula (I-B-ii)
wherein:
wherein indicates a single or double bond, and wherein the C6-C14 aryl and 5-to-10-membered heteroaryl are optionally substituted with 1-5 R6 groups;
In some embodiments, R3 is C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-CN, C1-C2 alkylene-(C3-C6 cycloalkyl), C1-C2 alkylene-O—(C1-C2 alkylene)-NR4R5, or C1-C2 alkylene-(C4-C6 heterocycloalkyl), wherein each heterocyclic nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl or C4-C6 heterocycloalkyl, wherein the R4 and R5 of R3 are independently —H or C1-C3 alkyl, and the R4′ and R5′ of R3 taken together with the nitrogen atom to which they are attached form a 4-to-6-membered heterocyclic ring, wherein the heterocyclic ring optionally contains an additional 1-2 heteroatoms selected from the group consisting of N and O, and wherein each additional nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl.
In some embodiments,
In some embodiments, each R6 is independently halogen or —OR7; and
In some embodiments of the compound of formula (I-B), formula (I-B-i) or formula (I-B-ii), R3 is C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkylene-NR4R5, C1-C3 alkylene-NR4′R5′, C1-C3 alkylene-CN, C1-C2 alkylene-(C3-C6 cycloalkyl), C1-C2 alkylene-O—(C1-C2 alkylene)-NR4R5, or C1-C2 alkylene-(C4-C6 heterocycloalkyl), wherein each heterocyclic nitrogen atom, if present, is independently optionally substituted with C1-C3 alkyl or C4-C6 heterocycloalkyl,
In some embodiments, R3 is
In some embodiments, each R6 is independently —F, —OCH3, or cyclopropyloxy. In some embodiments, R8 is H.
In some embodiments, provided is a compound selected from the compounds in Table 1, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing.
Although certain compounds described in Table 1 are presented as specific stereoisomers and/or in a non-stereochemical form, it is understood that any or all non-stereochemical forms and any or all stereochemical forms, including any enantiomeric or diastereomeric forms, and any tautomers or other forms of any of the compounds of Table 1 are herein described. In some embodiments, the compound described herein is selected from Compound Nos. 1-259.
This disclosure also includes all salts, such as pharmaceutically acceptable salts, of compounds referred to herein. This disclosure also includes any or all of the stereochemical forms, including any enantiomeric or diastereomeric forms, and any tautomers or other forms, such as N-oxides, solvates, hydrates, or isotopomers, of the compounds described. The present disclosure also includes co-crystals of the compounds described herein. Unless stereochemistry is explicitly indicated in a chemical structure or name, the structure or name is intended to embrace all possible stereoisomers of a compound depicted. In addition, where a specific stereochemical form is depicted, it is understood that other stereochemical forms are also embraced by the invention. All forms of the compounds are also embraced by the invention, such as crystalline or non-crystalline forms of the compounds. Compositions comprising a compound of the invention are also intended, such as a composition of substantially pure compound, including a specific stereochemical form thereof. Compositions comprising a mixture of compounds of the invention in any ratio are also embraced by the invention, including mixtures of two or more stereochemical forms of a compound of the invention in any ratio, such that racemic, non-racemic, enantioenriched and scalemic mixtures of a compound are embraced.
In the descriptions herein, it is understood that every description, variation, embodiment, or aspect of a moiety can be combined with every description, variation, embodiment, or aspect of other moieties the same as if each and every combination of descriptions is specifically and individually listed. For example, every description, variation, embodiment, or aspect provided herein with respect to R0 of formula (I) may be combined with every description, variation, embodiment, or aspect of X, m, R1, R2, R3, R4, R4′, R5, R5′, R6, R7, and/or R1, the same as if each and every combination were specifically and individually listed. It is also understood that all descriptions, variations, embodiments or aspects of formula (I), where applicable, apply equally to other formulae detailed herein, and are equally described, the same as if each and every description, variation, embodiment or aspect were separately and individually listed for all formulae. For example, all descriptions, variations, embodiments, or aspects of formula (I), where applicable, apply equally to any of formulae (I-A), (I-A-i), (I-B), (I-B-i) and (I-B-ii) detailed herein, and are equally described, the same as if each and every description, variation, embodiment or aspect were separately and individually listed for all formulae.
The compounds of the present disclosure may be prepared by a number of processes as generally described below and more specifically in the Examples hereinafter (such as the schemes provided in the Examples below). In the following process descriptions, the symbols when used in the formulae depicted are to be understood to represent those groups described above in relation to the formulae herein.
The intermediates described in the following preparations may contain a number of nitrogen, hydroxy, and acid protecting groups such as esters. The variable protecting group may be the same or different in each occurrence depending on the particular reaction conditions and the particular transformations to be performed. The protection and deprotection conditions are well known to the skilled artisan and are described in the literature. See e.g., Greene and Wuts, Protective Groups in Organic Synthesis, (T. Greene and P. Wuts, eds., 2d ed. 1991).
Certain stereochemical centers have been left unspecified and certain substituents have been eliminated in the following schemes for the sake of clarity and are not intended to limit the teaching of the schemes in any way. Furthermore, individual isomers, enantiomers, and diastereomers may be separated or resolved by one of ordinary skill in the art at any convenient point in the synthesis of compounds of the invention, by methods such as selective crystallization techniques or chiral chromatography (See e.g., J. Jacques, et al., “Enantiomers, Racemates, and Resolutions”, John Wiley and Sons, Inc., 1981, and E. L. Eliel and S. H. Wilen, “Stereochemistry of Organic Compounds”, Wiley-Interscience, 1994).
The compounds of the present invention, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, may be prepared by a variety of procedures known in the art, some of which are illustrated in the Examples below. The specific synthetic steps for each of the routes described may be combined in different ways, to prepare compounds of the present disclosure, or salts thereof. The products of each step can be recovered by conventional methods well known in the art, including extraction, evaporation, precipitation, chromatography, filtration, trituration, and crystallization. The reagents and starting materials are readily available to one of ordinary skill in the art. Others may be made by standard techniques of organic and heterocyclic chemistry which are analogous to the syntheses of known structurally-similar compounds and the procedures described in the Examples which follow including any novel procedures.
Compounds of formula (I) can be prepared according to Scheme A, Scheme B, Scheme C, Scheme D, Scheme E, or Scheme F, wherein X, m, R1, R2, R3, R4, R4′, R5, R5′, R6, R7, and/or R8 are as defined for formula (I) or any applicable variation thereof as detailed herein.
Compounds of formula (I-A-i) may be prepared according to the general synthetic scheme shown in Scheme A, Parts I-III. In Scheme A, Part I, 7-azaindole compounds of general formula A-i-a, wherein LG1 is a leaving group (such as chloro or bromo), are protected at the secondary nitrogen with a protecting group P1 (e.g., with a Boc- or SEM-group) to give the compounds of general formula A-i-b. A suitable leaving group LG2 (such as iodo) is added to the protected compounds of general formula A-i-b in preparation for the installation of the R2 moiety. The resulting compounds of general formula A-i-c are further subjected to reactions to introduce substituent R8′ onto the 7-azaindole core as required to produce the intermediate compounds of general formula A-i-e. As shown in Scheme A, R8′ has the same definition as R8 as described herein, except that R8′ does not include hydrogen.
In Part II of Scheme A, the R2 aryl or heteroaryl moiety is added to the compounds of general formula A-i-e, at the position occupied by LG2, to yield the further intermediate compounds of general formula A-i-i. The installation of the R2 moiety may be achieved, for example, by two routes as shown above. In the first route, the compounds of general formula A-i-e are reacted with a suitable boronic acid derivative comprising the desired R2 group A-i-f, wherein RA and RB are independently selected from the group consisting of halogen, OH, and O—(C1-C6 alkyl), or RA and RB are taken together with the boron atom to which they are attached to form a 5-10 membered heterocycle, to give the intermediate compounds of formula A-i-i. In the second route, the compounds of formula A-i-e are directly reacted with boronic acid or a derivative thereof, wherein Rc is a suitable leaving group (such as O—C1-C3 alkyl, or another boronic acid or derivative thereof, i.e., in a diboron compound), to give the 7-azaindolyl-boronate compounds of formula A-i-g. The resulting boronate compounds are further reacted with an R2-containing substrate (A-i-h) to give compounds of formula A-i-i.
In Part III of Scheme A, the compounds of formula A-i-i are reacted with suitable cyclopropanecarboxamides of formula A-i-j to provide compounds of formula I-A-i.
Similar to Scheme A, Scheme B describes the preparation of compounds of formula (I-A-ii) having a 7-azaindazolyl core. Compounds of formula (I-A-ii) may be prepared according to the general synthetic scheme shown in Scheme B, Parts I-III. In Scheme B, Part I, 7-azaindazole compounds of general formula A-ii-a, wherein LG1 is a leaving group (such as chloro or bromo), are protected at the secondary nitrogen with a protecting group P1 (e.g., with a Boc- or SEM-group) to give the compounds of general formula A-ii-b. A suitable leaving group LG2 (such as iodo) is added to the protected compounds of general formula A-ii-b in preparation for the installation of the R2 moiety.
In Part II of Scheme B, the R2 aryl or heteroaryl moiety is added to the compounds of general formula A-ii-c, at the position occupied by LG2, to yield the further intermediate compounds of general formula A-ii-g. The installation of the R2 moiety may be achieved, for example, by two routes as shown above. In the first route, the compounds of general formula A-ii-c are reacted with a suitable boronic acid derivative comprising the desired R2 group A-ii-d, wherein RA and RB are independently selected from the group consisting of halogen, OH, and O—(C1-C6 alkyl), or RA and RB are taken together with the boron atom to which they are attached to form a 5-10 membered heterocycle, to give the intermediate compounds of formula A-ii-g. In the second route, the compounds of formula A-ii-c are directly reacted with boronic acid or a derivative thereof, wherein Rc is a suitable leaving group (such as O—C1-C3 alkyl, or another boronic acid or derivative thereof, i.e., in a diboron compound), to give the 7-azaindazolyl-boronate compounds of formula A-ii-e. The resulting boronate compounds are further reacted with an R2-containing substrate (A-ii-f) to give compounds of formula A-ii-g.
In Part III of Scheme B, the compounds of formula A-ii-g are reacted with suitable cyclopropanecarboxamides of formula A-ii-h to provide compounds of formula I-A-ii.
Compounds of formula (I-B) may also be prepared according to the general synthetic schemes shown in Scheme C and Scheme D. In Scheme C, the 7-azaindolyl compounds of formula A-i-i obtained from Scheme A (Parts I and II) are reacted with suitable urea compounds of formula B-i-j to provide compounds of formula I-B-i. In Scheme D, the 7-azaindazolyl compounds of formula A-ii-g obtained from Scheme B (Parts I and II) are reacted with suitable urea compounds of formula B-ii-j to provide compounds of formula I-B-ii.
Alternatively, the addition of the cyclopropylcarboxamidyl or urea moiety as described in Scheme A (Part III), Scheme B (Part III), Scheme C and Scheme D may be substituted with the methods as described in Scheme E, Scheme F, Scheme G and Scheme H, respectively.
In Scheme E, the intermediate compounds of general formula A-i-i are aminated (such as with diphenylmethanimine) to give the compounds of general formula A-i-k. The resulting compounds of general formula A-i-k are subsequently reacted with a cyclopropanecarboxylic acid or derivative thereof A-i-l (wherein LG3 may be —OH, Cl—, —O—C1-C6 alkyl, etc.) to give the desired compounds of formula (I-A-i). Similarly, in Scheme F, the intermediate compounds of general formula A-ii-g are aminated (such as with diphenylmethanimine) to give the compounds of general formula A-ii-i. The resulting compounds of general formula A-ii-i are subsequently reacted with a cyclopropanecarboxylic acid or derivative thereof A-ii-j (wherein LG3 may be —OH, Cl—, —O—C1-C6 alkyl, etc.) to give the desired compounds of formula (I-A-ii).
Compounds of general formula (I-B) may also be prepared from the compounds of general formulae A-i-k and A-ii-i as described in Schemes G and F. In Scheme G, the compounds of general formula A-i-k are reacted with carboxylic acid derivatives (e.g., phenyl carbonylchloridate and R3-containing free amines B-i-m in successive steps to give the desired urea compounds of general formula I-B-i. Similarly, in Scheme H, the compounds of general formula A-ii-i are reacted with carboxylic acid derivatives and R3-containing free amines B-ii-m in successive steps to give the desired urea compounds of general formula I-Bii.
Any of the compounds described herein may be formulated as a pharmaceutically acceptable composition.
Pharmaceutical compositions of any of the compounds detailed herein are embraced by this disclosure. Thus, the present disclosure includes pharmaceutical compositions comprising a compound as detailed herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, and a pharmaceutically acceptable carrier or excipient. In one aspect, the pharmaceutically acceptable salt is an acid addition salt, such as a salt formed with an inorganic or organic acid. Pharmaceutical compositions may take a form suitable for oral, buccal, parenteral, nasal, topical or rectal administration or a form suitable for administration by inhalation.
A compound as detailed herein may in one aspect be in a purified form and compositions comprising a compound in purified forms are detailed herein. Compositions comprising a compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, as detailed herein are provided, such as compositions of substantially pure compounds. In some embodiments, a composition containing a compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, as detailed herein is in substantially pure form. In one variation, “substantially pure” intends a composition that contains no more than 35% impurity, wherein the impurity denotes a compound other than the compound comprising the majority of the composition or a salt thereof. For example, a composition of a substantially pure compound selected from a compound of Table 1 intends a composition that contains no more than 35% impurity, wherein the impurity denotes a compound other than the compound of Table 1. In one variation, a composition of substantially pure compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, is provided wherein the composition contains no more than 25% impurity. In another variation, a composition of substantially pure compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, is provided wherein the composition contains or no more than 20% impurity. In still another variation, a composition of substantially pure compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, is provided wherein the composition contains or no more than 10% impurity. In a further variation, a composition of substantially pure compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, is provided wherein the composition contains no more than 5% impurity. In another variation, a composition of substantially pure compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, is provided wherein the composition contains no more than 3% impurity. In still another variation, a composition of substantially pure compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, is provided wherein the composition contains no more than 1% impurity. In a further variation, a composition of substantially pure compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, is provided wherein the composition contains no more than 0.5% impurity. In yet other variations, a composition of substantially pure compound means that the composition contains no more than 15%, no more than 10%, no more than 5%, no more than 3%, or no more than 1% impurity, which impurity may be the compound in a different stereochemical form. For instance, and without limitation, a composition of substantially pure (S) compound means that the composition contains no more than 15% or no more than 10% or no more than 5% or no more than 3% or no more than 1% of the (R) form of the compound.
In one variation, the compounds herein are synthetic compounds prepared for administration to an individual. In another variation, compositions are provided containing a compound in substantially pure form. In another variation, the present disclosure embraces pharmaceutical compositions comprising a compound detailed herein and a pharmaceutically acceptable carrier. In another variation, methods of administering a compound are provided. The purified forms, pharmaceutical compositions and methods of administering the compounds are suitable for any compound or form thereof detailed herein. In some embodiments, the compounds and compositions as provided herein are sterile. Methods for sterilization known in the art may be suitable for any compounds or form thereof and compositions thereof as detailed herein.
A compound detailed herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, may be formulated for any available delivery route, including an oral, mucosal (e.g., nasal, sublingual, vaginal, buccal or rectal), parenteral (e.g., intramuscular, subcutaneous or intravenous), topical or transdermal delivery form. A compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, may be formulated with suitable carriers to provide delivery forms that include, but are not limited to, tablets, caplets, capsules (such as hard gelatin capsules or soft elastic gelatin capsules), cachets, troches, lozenges, gums, dispersions, suppositories, ointments, cataplasms (poultices), pastes, powders, dressings, creams, solutions, patches, aerosols (e.g., nasal spray or inhalers), gels, suspensions (e.g., aqueous or non-aqueous liquid suspensions, oil-in-water emulsions or water-in-oil liquid emulsions), solutions and elixirs.
A compound detailed herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, can be used in the preparation of a formulation, such as a pharmaceutical formulation, by combining the compound or compounds, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, with a pharmaceutically acceptable carrier. Depending on the therapeutic form of the system (e.g., transdermal patch vs. oral tablet), the carrier may be in various forms. In addition, pharmaceutical formulations may contain preservatives, solubilizers, stabilizers, re-wetting agents, emulgators, sweeteners, dyes, adjusters, and salts for the adjustment of osmotic pressure, buffers, coating agents or antioxidants. Formulations comprising the compound may also contain other substances which have valuable therapeutic properties. Pharmaceutical formulations may be prepared by known pharmaceutical methods. Suitable formulations can be found, e.g., in Remington's Pharmaceutical Sciences, Mack Publishing Company, Philadelphia, PA, 20th ed. (2000), which is incorporated herein by reference.
A compound detailed herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, may be administered to individuals in a form of generally accepted oral compositions, such as tablets, coated tablets, and gel capsules in a hard or in soft shell, emulsions or suspensions. Examples of carriers, which may be used for the preparation of such compositions, are lactose, corn starch or its derivatives, talc, stearate or its salts, etc. Acceptable carriers for gel capsules with soft shell are, for instance, plant oils, wax, fats, semisolid and liquid poly-ols, and so on. In addition, pharmaceutical formulations may contain preservatives, solubilizers, stabilizers, re-wetting agents, emulgators, sweeteners, dyes, adjusters, and salts for the adjustment of osmotic pressure, buffers, coating agents or antioxidants.
Any of the compounds, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, described herein can be formulated in a tablet in any dosage form described, for example, a compound as described herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, can be formulated as a 10 mg tablet.
Compositions comprising a compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, provided herein are also described. In one variation, the composition comprises a compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, and a pharmaceutically acceptable carrier or excipient. In another variation, a composition of substantially pure compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, is provided. In some embodiments, the composition is for use as a human or veterinary medicament. In some embodiments, the composition is for use in a method described herein. In some embodiments, the composition is for use in the treatment of a disease or disorder described herein.
Compositions formulated for co-administration of a compound provided herein and one or more additional pharmaceutical agents are also described. The co-administration can be simultaneous or sequential in any order. A compound provided herein may be formulated for co-administration with the one or more additional pharmaceutical agents in the same dosage form (e.g., single tablet or single i.v.) or separate dosage forms (e.g., two separate tablets, two separate i.v., or one tablet and one i.v.). Furthermore, co-administration can be, for example, 1) concurrent delivery, through the same route of delivery (e.g., tablet or i.v.), 2) sequential delivery on the same day, through the same route or different routes of delivery, or 3) delivery on different days, through the same route or different routes of delivery.
Compounds and compositions detailed herein, such as a pharmaceutical composition containing a compound of formula (I) or any variation thereof provided herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, and a pharmaceutically acceptable carrier or excipient, may be used in methods of administration and treatment as provided herein. The compounds and compositions may also be used in in vitro methods, such as in vitro methods of administering a compound or composition to cells for screening purposes and/or for conducting quality control assays.
In one aspect, provided herein is a method of inhibiting Bcr-Abl tyrosine kinase enzymatic activity, comprising contacting an effective amount of a compound or composition provided herein, to the Bcr-Abl tyrosine kinase. In some embodiments, provided herein is a method of inhibiting Bcr-Abl tyrosine kinase in a cell, comprising administering an effective amount of a compound or composition of the disclosure to the cell. In some embodiments, provided herein is a method of inhibiting Bcr-Abl tyrosine kinase in an individual in need thereof, comprising administering an effective amount of a compound or composition of the disclosure to the individual. In some variations, the compounds provided herein are selective for inhibiting Bcr-Abl tyrosine kinase. As such, in some embodiments, provided herein is a method of selectively inhibiting Bcr-Abl tyrosine kinase, as compared to other tyrosine kinases, including but not limited to c-KIT, FGFR, PDGFR, SRC, CSFR1, or VEGFR.
The compounds and compositions described herein may be used in a method of treating a disease or disorder mediated by Bcr-Abl tyrosine kinase activity. In some embodiments, the compound or composition is administered according to a dosage described herein.
In some embodiments, provided herein is a method for treating a disease or disorder mediated by Bcr-Abl tyrosine kinase activity comprising administering to an individual in need of treatment an effective amount of a compound of formula (I) or any variation thereof, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing. In some embodiments, the disease or disorder is a cancer mediated by Bcr-Abl tyrosine kinase activity. In some embodiments, the disease or disorder is chronic myeloid leukemia (CML), acute myeloid leukemia (AML), or acute lymphoblastic leukemia (ALL). In some embodiments, the disease or disorder is a cancer, such as leukemia. In some variations, the cancer is chronic myeloid leukemia (CML), Philadelphia-positive acute lymphoblastic leukemia (Ph+ALL), acute myelogenous leukemia (AML), or mixed phenotype acute leukemia.
In certain embodiments, the leukemia is chronic myeloid leukemia. Chronic myeloid leukemia may be characterized by the state of disease progression, as determined by blast cells. In still further embodiments, the chronic myeloid leukemia is chronic phase CML, accelerated phase CML, or blastic phase CML. In some embodiments, the chronic myeloid leukemia is refractory chronic myeloid leukemia.
In some embodiments, the disease or disorder mediated by Bcr-Abl tyrosine kinase activity is refractory or resistant to first-line treatment, second-line treatment, and/or third-line treatment. In certain embodiments, the condition mediated by Bcr-Abl tyrosine kinase activity is refractory or resistant to treatment with one or more Bcr-Abl tyrosine kinase inhibitors selected from the group consisting of imatinib, nilotinib, dasatinib, bafetinib, bosutinib, radotinib, asciminib, and ponatinib. First-line treatment as described herein includes the use of imatinib; second- and third-line treatments as described herein include the use of nilotinib, dasatinib, bafetinib, bosutinib, radotinib, asciminib, and/or ponatinib. In some variations of the foregoing, the chronic myeloid leukemia is refractory chronic myeloid leukemia.
Resistant subtypes of Bcr-Abl tyrosine kinase-mediated diseases or disorders may be associated with any number of Bcr-Abl dependent or Bcr-Abl independent resistance mechanisms. In some embodiments wherein the disease or disorder mediated by Bcr-Abl tyrosine kinase activity is refractory to treatment, the disease or disorder is characterized as being associated with one or more Bcr-Abl dependent resistance mechanisms. Bcr-Abl dependent resistance mechanisms include, but are not limited to, one or more point mutations at positions M244, L248, G250, G250, Q252, Q252, Y253, Y253, E255, E255, D276, F311, T315, T315, F317, F317, M343, M351, E355, F359, F359, V379, F382, L387, H396, H396, S417, E459, F486, or T315 in the Bcr-Abl tyrosine kinase. In certain variations, the refractory disease or disorder mediated by Bcr-Abl tyrosine kinase is associated with one or more specific point mutations in the Bcr-Abl tyrosine kinase selected from the group consisting of: M244V, L248V, G250E, G250A, Q252H, Q252R, Y253F, Y253H, E255K, E255V, D276G, F311L, T315N, T315A, F317V, F317L, M343T, M351T, E355G, F359A, F359V, V379I, F382L, L387M, H396P, H396R, S417Y, E459K, F486S, and T315I. In certain embodiments, the refractory disease or disorder mediated by Bcr-Abl tyrosine kinase is associated with a T315I mutation. In still further embodiments, the refractory disease or disorder mediated by Bcr-Abl tyrosine kinase is associated with a T315I mutation at the onset of treatment and I315M mutation following ponatinib. In other embodiments, the refractory disease or disorder mediated by Bcr-Abl tyrosine kinase is associated with one or more P-loop mutations (M244V, G250E, Q252H, Y253H/F, E255K/V).
In some embodiments, provided is a method for treating cancer in an individual in need thereof, comprising administering to the individual an effective amount of a compound of formula (I), or any variation thereof as described herein. In some embodiments, the cancer is leukemia. In some embodiments, the cancer is chronic myeloid leukemia (CML), Philadelphia-positive acute lymphoblastic leukemia (Ph+ALL), acute myelogenous leukemia (AML), or mixed phenotype acute leukemia. In some embodiments, the cancer is chronic myeloid leukemia (CML). In certain embodiments, the leukemia is chronic myeloid leukemia. In still further embodiments, the chronic myeloid leukemia is refractory chronic myeloid leukemia. In certain embodiments of the foregoing, the chronic myeloid leukemia is refractory chronic myeloid leukemia associated with a T315I mutation.
In one aspect, provided herein is a method of treating cancer in an individual in need thereof, wherein modulation of Bcr-Abl tyrosine kinase activity inhibits or ameliorates the pathology and/or symptomology of the cancer, comprising administering to the individual a therapeutically effective amount of a compound or composition provided herein. In one embodiment, provided herein is a method of treating cancer, wherein modulation of Bcr-Abl tyrosine kinase activity inhibits the pathology and/or symptomology of the cancer, in an individual, comprising administering to the individual a therapeutically effective amount of a compound or composition provided herein. In one embodiment, provided herein is a method of treating a cancer, wherein modulation of Bcr-Abl tyrosine kinase activity ameliorates the pathology and/or symptomology of the cancer, in an individual, comprising administering to the individual a therapeutically effective amount of a compound or composition provided herein.
In another aspect, provided herein is a method of preventing cancer, wherein modulation of Bcr-Abl tyrosine kinase activity prevents the pathology and/or symptomology of the cancer, in an individual, comprising administering to the individual a therapeutically effective amount of a compound or composition provided herein. In another aspect, provided herein is a method of delaying the onset and/or development of a cancer that is mediated by Bcr-Abl tyrosine kinase activity in an individual (such as a human) who is at risk for developing the cancer. It is appreciated that delayed development may encompass prevention in the event the individual does not develop the cancer.
In one aspect, provided herein is a method of delaying the onset and/or development of cancer in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound or composition provided herein. In some embodiments, the cancer is a leukemia. In certain embodiments, the cancer is chronic myeloid leukemia (CML), Philadelphia-positive acute lymphoblastic leukemia (Ph+ALL), acute myelogenous leukemia (AML), or mixed phenotype acute leukemia. In some embodiments, the cancer is chronic myeloid leukemia. In still further embodiments, the chronic myeloid leukemia is refractory chronic myeloid leukemia. In still yet other embodiments, the chronic myeloid leukemia is refractory chronic myeloid leukemia associated with a T315I mutation. In one aspect, provided herein is a method of delaying the onset and/or development of chronic myeloid leukemia in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound or composition provided herein. In one variation, provided herein is a method of delaying the onset and/or development of refractory chronic myeloid in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound or composition provided herein. In one variation, provided herein is a method of delaying the onset and/or development of refractory chronic myeloid leukemia associated with a T315I mutation in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound or composition provided herein.
In one aspect, provided herein is a compound of formula (I) or any variation thereof, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, for use in therapy. In some embodiments, provided herein is a compound of formula (I) or any variation thereof, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, or pharmaceutical composition comprising such compound, for use in the treatment of cancer. In some embodiments, provided is a compound of formula (I) or any variation thereof, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, or a pharmaceutical composition comprising such compound, for use in the treatment of chronic myeloid leukemia (CML), Philadelphia-positive acute lymphoblastic leukemia (Ph+ALL), acute myelogenous leukemia (AML), or mixed phenotype acute leukemia. In some embodiments, provided is a compound of formula (I) or any variation thereof, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, or a pharmaceutical composition comprising such compound, for use in the treatment of chronic myeloid leukemia (CML). In some embodiments, provided is a compound of formula (I) or any variation thereof, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, or a pharmaceutical composition comprising such compound, for use in the treatment of refractory chronic myeloid leukemia (CML). In some embodiments, provided is a compound of formula (I) or any variation thereof, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, or a pharmaceutical composition comprising such compound, for use in the treatment of refractory chronic myeloid leukemia associated with a T315I mutation.
In another embodiment, provided herein is a compound of formula (I) or any variation thereof, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, for use in the manufacture of a medicament for the treatment of cancer. In another embodiment, provided herein is a compound of formula (I) or any variation thereof, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, for use in the manufacture of a medicament for the treatment of chronic myeloid leukemia (CML), Philadelphia-positive acute lymphoblastic leukemia (Ph+ALL), acute myelogenous leukemia (AML), or mixed phenotype acute leukemia. In some embodiments, the medicament is for the treatment of chronic myeloid leukemia. In some embodiments, the medicament is for the treatment of refractory chronic myeloid leukemia. In some embodiments, the medicament is for the treatment of refractory chronic myeloid leukemia associated with a T315I mutation.
In some embodiments, the individual is a mammal. In some embodiments, the individual is a primate, dog, cat, rabbit, or rodent. In some embodiments, the individual is a primate. In some embodiments, the individual is a human. In some embodiments, the human is at least about or is about any of 18, 21, 30, 50, 60, 65, 70, 75, 80, or 85 years old. In some embodiments, the human is a child. In some embodiments, the human is less than about or about any of 21, 18, 15, 10, 5, 4, 3, 2, or 1 years old.
In some embodiments, the method further comprises administering one or more additional pharmaceutical agents. In some embodiments, the method further comprises administering radiation. In some embodiments, the method further comprises administering one or more additional pharmaceutical agents, including anti-microtubular therapies (e.g. paclitaxel, vincristine), topoisomerase inhibitors (e.g. adriamycin), alylating agents (e.g. busulfan, cyclophosphamide), nucleotide synthesis inhibitors (hyroxyurea), DNA synthesis inhibitors (e.g. cytarabine), protein synthesis inhibitors (e.g. omacetaxine), developmental signaling pathway inhibitors (e.g. sonidegib, Hedgehog pathway), pro-apoptotic agents (e.g. venetoclax), Abl myristoyl-pocket binding inhibitors (e.g. asciminib), MEK1/2 inhibitors (e.g. trametinib, binimetinib), AKT inhibitors (e.g. ipatasertib), PI3K inhibitors (e.g. apelisib) and radiation.
The dose of a compound described herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, administered to an individual (such as a human) may vary with the particular compound or salt thereof, the method of administration, and the particular cancer, such as type and stage of cancer, being treated. In some embodiments, the amount of the compound, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, is a therapeutically effective amount.
The compounds provided herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, may be administered to an individual via various routes, including, e.g., intravenous, intramuscular, subcutaneous, oral, and transdermal.
The effective amount of the compound may in one aspect be a dose of between about 0.01 and about 100 mg/kg. Effective amounts or doses of the compounds of the present disclosure may be ascertained by routine methods, such as modeling, dose escalation, or clinical trials, taking into account routine factors, e.g., the mode or route of administration or drug delivery, the pharmacokinetics of the agent, the severity and course of the disease to be treated, the subject's health status, condition, and weight. An exemplary dose is in the range of about from about 0.7 mg to 7 g daily, or about 7 mg to 350 mg daily, or about 350 mg to 1.75 g daily, or about 1.75 to 7 g daily.
Any of the methods provided herein may in one aspect comprise administering to an individual a pharmaceutical composition that contains an effective amount of a compound provided herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, and a pharmaceutically acceptable excipient.
A compound or composition provided herein may be administered to an individual in accordance with an effective dosing regimen for a desired period of time or duration, such as at least about one month, at least about 2 months, at least about 3 months, at least about 6 months, or at least about 12 months or longer, which in some variations may be for the duration of the individual's life. In one variation, the compound is administered on a daily or intermittent schedule. The compound can be administered to an individual continuously (for example, at least once daily) over a period of time. The dosing frequency can also be less than once daily, e.g., about a once weekly dosing. The dosing frequency can be more than once daily, e.g., twice or three times daily. The dosing frequency can also be intermittent, including a ‘drug holiday’ (e.g., once daily dosing for 7 days followed by no doses for 7 days, repeated for any 14 day time period, such as about 2 months, about 4 months, about 6 months or more). Any of the dosing frequencies can employ any of the compounds described herein together with any of the dosages described herein.
The present disclosure further provides articles of manufacture comprising a compound described herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, a composition described herein, or one or more unit dosages described herein in suitable packaging. In certain embodiments, the article of manufacture is for use in any of the methods described herein. Suitable packaging is known in the art and includes, for example, vials, vessels, ampules, bottles, jars, flexible packaging and the like. An article of manufacture may further be sterilized and/or sealed.
The present disclosure further provides kits for carrying out the methods of the present disclosure, which comprises one or more compounds described herein or a composition comprising a compound described herein. The kits may employ any of the compounds disclosed herein. In one variation, the kit employs a compound described herein, or a pharmaceutically acceptable salt, solvate, hydrate, or co-crystal thereof, or a mixture of any of the foregoing, thereof. The kits may be used for any one or more of the uses described herein, and, accordingly, may contain instructions for the treatment of any disease or described herein, for example for the treatment of cancer, including chronic myeloid leukemia (CML), Philadelphia-positive acute lymphoblastic leukemia (Ph+ALL), acute myelogenous leukemia (AML), or mixed phenotype acute leukemia. In some embodiments, the cancer is chronic myeloid leukemia. In some embodiments, the cancer is refractory chronic myeloid leukemia. In certain embodiments of the foregoing, the cancer is refractory chronic myeloid leukemia associated with a T315I mutation.
The kits optionally further comprise a container comprising one or more additional pharmaceutical agents and which kits further comprise instructions on or in the package insert for treating the subject with an effective amount of the one or more additional pharmaceutical agents.
Kits generally comprise suitable packaging. The kits may comprise one or more containers comprising any compound described herein. Each component (if there is more than one component) can be packaged in separate containers or some components can be combined in one container where cross-reactivity and shelf life permit.
The kits may be in unit dosage forms, bulk packages (e.g., multi-dose packages) or sub-unit doses. For example, kits may be provided that contain sufficient dosages of a compound as disclosed herein and/or an additional pharmaceutically active compound useful for a disease detailed herein to provide effective treatment of an individual for an extended period, such as any of a week, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 7 months, 8 months, 9 months, or more. Kits may also include multiple unit doses of the compounds and instructions for use and be packaged in quantities sufficient for storage and use in pharmacies (e.g., hospital pharmacies and compounding pharmacies).
The kits may optionally include a set of instructions, generally written instructions, although electronic storage media (e.g., magnetic diskette or optical disk) containing instructions are also acceptable, relating to the use of component(s) of the methods of the present disclosure. The instructions included with the kit generally include information as to the components and their administration to an individual.
It is understood that the present disclosure has been made only by way of example, and that numerous changes in the combination and arrangement of parts can be resorted to by those skilled in the art without departing from the spirit and scope of present disclosure.
The chemical reactions in the Examples described can be readily adapted to prepare a number of other compounds disclosed herein, and alternative methods for preparing the compounds of this disclosure are deemed to be within the scope of this disclosure. For example, the synthesis of non-exemplified compounds according to the present disclosure can be successfully performed by modifications apparent to those skilled in the art, e.g., by appropriately protecting interfering groups, by utilizing other suitable reagents known in the art other than those described, or by making routine modifications of reaction conditions, reagents, and starting materials. Alternatively, other reactions disclosed herein or known in the art will be recognized as having applicability for preparing other compounds of the present disclosure.
Abbreviations used in the Examples include the following: ACN: acetonitrile; Brettphos: 2-(dicyclohexylphosphino)3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl; dppf: 1,1′-ferrocenediyl-bis(diphenylphosphine); DCM: dichloromethane; DIAD: diisopropylazodicarboxylate; DIEA: N,N-diisopropylethylamine; DMAP: 4-dimethylaminopyridine; DMF: dimethylformamide; DMF-DMA: N,N-dimethylformamide dimethylacetal; DMSO: dimethyl sulfoxide; EDA; ethylenediamine; EtOAc: ethyl acetate; EtOH: ethanol or ethyl alcohol; F-TEDA-BF4: 1-Chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane bis(tetrafluoroborate); 1H NMR: proton nuclear magnetic resonance; HATU: 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, Hexafluorophosphate Azabenzotriazole Tetramethyl Uronium; LCMS: liquid chromatography-mass spectrometry; LDA: lithium diisopropylamide; LiHMDS: lithium hexamethyldisilazide; MeOH: methanol or methyl alcohol; NBS: N-bromosuccinimide; NIS: N-iodosuccinimide; NNMP: N-methyl-2-pyrrolidone; OAc: acetate; Py: pyridine; TBAB: tetra-n-butylammonium bromide; TBAF: tetra-n-butylammonium fluoride; TEA: triethylamine; TFA: trifluoroacetic acid; THF: tetrahydrofuran; and TLC: thin-layer chromatography.
To a solution of 6-chloro-1H-pyrrolo[2,3-b]pyridine (500.0 mg, 3.28 mmol) in DMF (5.0 mL) was added Br2 (549.9 mg, 3.44 mmol) dropwise at 0° C. under N2. The resulting mixture was stirred at 0° C. for 3 h under N2. After the reaction was completed, the reaction was quenched with aq·NaHSO3 at 10° C. The pH value of the mixture was adjusted to 8 with aq·NaHCO3. The resulting mixture was filtered. The solid was washed with H2O, collected and dried to afford 3-bromo-6-chloro-1H-pyrrolo[2,3-b]pyridine (620.0 mg, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=230.9.
To a solution of 3-bromo-6-chloro-1H-pyrrolo[2,3-b]pyridine (620.0 mg, crude) in CH2Cl2 (10.0 mL) was added Boc2O (759.9 mg, 3.48 mmol) and TEA (758.9 mg, 7.50 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (85/15, v/v) to afford tert-butyl 3-bromo-6-chloro-1H-pyrrolo[2,3-b]pyridine-1-carboxylate (850.0 mg, 96%) as a white solid. LCMS (ESI, m/z): [M+H]+=331.0.
To a solution of tert-butyl 3-bromo-6-chloro-1H-pyrrolo[2,3-b]pyridine-1-carboxylate (500.0 mg, 1.51 mmol) in 1,4-dioxane/H2O (20.0/2.0 mL) was added (2-methoxyphenyl)boronic acid (275.0 mg, 1.81 mmol), K2CO3 (625.2 mg, 4.52 mmol) and Pd(dppf)Cl2 (110.3 mg, 0.15 mmol). The resulting mixture was stirred at 100° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (83/17, v/v) to afford 6-chloro-3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridine (190.0 mg, 49%) as a white solid. LCMS (ESI, m/z): [M+H]+=259.1.
To a stirred solution of 6-chloro-3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridine (140.0 mg, 0.54 mmol) in 1,4-dioxane (15.0 mL) was added cyclopropanecarboxamide (184.2 mg, 2.17 mmol), BrettPhos (58.1 mg, 0.11 mmol), Cs2CO3 (529.0 mg, 1.62 mmol) and BrettPhos Pd G3 (49.1 mg, 0.05 mmol). The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (95/5, v/v) and then purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 27% B to 57% B in 7 min; 254 nm) to afford N-(3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (14.6 mg, 9%) as a white solid. LCMS (ESI, m/z): [M+H]+=308.2. 1H NMR (300 MHz, DMSO-d6): δ 11.55 (s, 1H), 10.58 (s, 1H), 7.98-7.87 (m, 2H), 7.56-7.52 (m, 2H), 7.35-7.19 (m, 1H), 7.11 (d, J=8.1 Hz, 1H), 7.07-6.92 (m, 1H), 3.81 (s, 3H), 2.11-1.98 (m, 1H), 0.97-0.73 (m, 4H).
To a solution of 3-bromo-6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (600.0 mg, 1.66 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added (2-methoxyphenyl)boronic acid (302.5 mg, 1.99 mmol), K2CO3 (687.7 mg, 4.98 mmol) and Pd(dppf)Cl2 (121.4 mg, 0.17 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 3 h under N2. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 6-chloro-3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (390.0 mg, 60%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=389.1.
To a solution of 6-chloro-3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (190.0 mg, 0.49 mmol) in 1,4-dioxane (10.0 mL) was added (1R,2R)-2-fluorocyclopropane-1-carboxamide (251.8 mg, 2.44 mmol), BrettPhos (52.4 mg, 0.10 mmol), Cs2CO3 (477.4 mg, 1.47 mmol) and BrettPhos Pd G3 (44.3 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (67/33, v/v) to afford (1R,2R)-2-fluoro-N-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (110.0 mg, 49%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=456.2.
To a solution of (1R,2R)-2-fluoro-N-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (100.0 mg, 0.22 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The reaction mixture was evaporated in vacuo. The residue was re-dissolved in ACN/H2O (5.0/1.0 mL). Then K2CO3 (380.7 mg, 2.76 mmol) was added to the mixture. The mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 39% B to 52% B in 7 min; 254 nm) to afford (1R,2R)-2-fluoro-N-(3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (14.0 mg, 20%) as a white solid. LCMS (ESI, m/z): [M+H]+=326.1. 1H NMR (300 MHz, DMSO-d6): δ 11.58 (s, 1H), 10.64 (s, 1H), 8.00 (d, J=8.7 Hz, 1H), 7.90 (d, J=8.7 Hz, 1H), 7.58-7.53 (m, 2H), 7.31-7.25 (m, 1H), 7.12 (d, J=7.5 Hz, 1H), 7.06-7.00 (m, 1H), 5.10-4.78 (m, 1H), 3.82 (s, 3H), 2.30-2.15 (m, 1H), 1.75-1.67 (m, 1H), 1.25-1.08 (m, 1H).
To a solution of 6-chloro-3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (190.0 mg, 0.49 mmol) in 1,4-dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (251.8 mg, 2.44 mmol), BrettPhos (52.4 mg, 0.10 mmol), Cs2CO3 (477.4 mg, 1.47 mmol) and BrettPhos Pd G3 (44.3 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (67/33, v/v) to afford (1S,2S)-2-fluoro-N-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (100.0 mg, 45%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=456.2.
To a solution of (1S,2S)-2-fluoro-N-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (100.0 mg, 0.22 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The reaction mixture was evaporated in vacuo. The residue was re-dissolved in ACN/H2O (5.0/1.0 mL). Then K2CO3 (380.7 mg, 2.76 mmol) was added to the mixture. The reaction mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 39% B to 52% B in 7 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (10.6 mg, 14%) as a light pink solid. LCMS (ESI, m/z): [M+H]+=326.2. 1H NMR (300 MHz, DMSO-d6): δ 11.57 (s, 1H), 10.63 (s, 1H), 8.00 (d, J=8.7 Hz, 1H), 7.89 (d, J=8.7 Hz, 1H), 7.57-7.52 (m, 2H), 7.30-7.24 (m, 1H), 7.12-7.09 (m, 1H), 7.05-7.00 (m, 1H), 5.05-4.80 (m, 1H), 3.82 (s, 3H), 2.27-2.21 (m, 1H), 1.70-1.61 (m, 1H), 1.19-1.12 (m, 1H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.83 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 4,4,5,5-tetramethyl-2-(2-methylphenyl)-1,3,2-dioxaborolane (217.0 mg, 0.99 mmol), K2CO3 (343.9 mg, 2.49 mmol) and Pd(dppf)Cl2 (121.4 mg, 0.17 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to afford 6-chloro-3-(2-methylphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (150.0 mg, 49%) as a white solid. LCMS (ESI, m/z): [M+H]+=373.1.
To a solution of 6-chloro-3-(2-methylphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (120.0 mg, 0.32 mmol) in dioxane (8.0 mL) was added cyclopropanecarboxamide (136.9 mg, 1.60 mmol), BrettPhos (69.0 mg, 0.13 mmol), Cs2CO3 (314.5 mg, 0.97 mmol) and BrettPhos Pd G3 (58.3 mg, 0.06 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford N-[3-(2-methylphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (120.0 mg, 88%) as a white solid. LCMS (ESI, m/z): [M+H]+=422.2.
To a solution of N-[3-(2-methylphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (100.0 mg, 0.24 mmol) in DCM (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in ACN/H2O (5.0/1.0 mL). Then K2CO3 (163.6 mg, 1.18 mmol) was added to the mixture. The mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 48% B to 58% B in 7 min; 254 nm) to afford N-[3-(2-methylphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (13.6 mg, 20%) as a white solid. LCMS (ESI, m/z): [M+H]+=292.1. 1H NMR (400 MHz, DMSO-d6): δ 11.59 (s, 1H), 11.60 (s, 1H), 7.89 (d, J=8.4 Hz, 1H), 7.78 (s, 1H), 7.42 (d, J=2.4 Hz, 1H), 7.37-7.31 (m, 2H), 7.25-7.22 (m, 2H), 2.31 (s, 3H), 2.06-2.02 (m, 1H), 0.82-0.80 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.83 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 2-fluorophenylboronic acid (139.2 mg, 1.01 mmol), K2CO3 (343.8 mg, 2.49 mmol) and Pd(dppf)Cl2 (60.7 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 3 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (93/7, v/v) to afford 6-chloro-3-(2-fluorophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (220.0 mg, 70%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=377.1.
To a solution of 6-chloro-3-(2-fluorophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (170.0 mg, 0.45 mmol) in 1,4-dioxane (5.0 mL) was added cyclopropanecarboxamide (115.1 mg, 1.35 mmol), Cs2CO3 (440.8 mg, 1.35 mmol), Brettphos (48.42 mg, 0.09 mmol) and Brettphos Pd G3 (40.9 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (83/17, v/v) to afford N-[3-(2-fluorophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (210.0 mg, 62%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=426.1.
To a solution of N-[3-(2-fluorophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (150.0 mg, 0.35 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL). The resulting mixture was stirred at room temperature for 1 h. The reaction mixture was evaporated in vacuo. The residue was re-dissolved in ACN/H2O (5.0/1.0 mL). Then K2CO3 (487.1 mg, 3.53 mmol) was added to the mixture. The mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 38% B to 68% B in 7 min; 254 nm) to afford N-[3-(2-fluorophenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (54.7 mg, 52%) as a white solid. LCMS (ESI, m/z): [M+H]+=296.2. 1H NMR (300 MHz, DMSO-d6): δ 11.80 (s, 1H), 10.65 (s, 1H), 8.08-8.05 (m, 1H), 7.95 (d, J=8.7 Hz, 1H), 7.76-7.70 (m, 1H), 7.66 (s, 1H), 7.34-7.26 (m, 3H), 2.08-2.03 (m, 1H), 0.86-0.76 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.83 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 2-chlorophenylboronic acid (155.6 mg, 0.99 mmol), K2CO3 (343.9 mg, 2.49 mmol) and Pd(dppf)Cl2 (60.7 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 3 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 6-chloro-3-(2-chlorophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (210.0 mg, 64%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=393.1.
To a solution of 6-chloro-3-(2-chlorophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (110.0 mg, 0.28 mmol) in 1,4-dioxane (3.0 mL) was added cyclopropanecarboxamide (23.8 mg, 0.28 mmol), BrettPhos (30.0 mg, 0.06 mmol), Cs2CO3 (273.3 mg, 0.38 mmol) and BrettPhos Pd G3 (25.4 mg, 0.03 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 3 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford N-[3-(2-chlorophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (105.0 mg, 85%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=442.2.
To a solution of N-[3-(2-chlorophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (105.0 mg, 0.24 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The mixture was evaporated in vacuo. The residue was re-dissolved with ACN/H2O (5.0/3.0 mL). Then K2CO3 (328.3 mg, 2.38 mmol) was added to the mixture. The mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep Phenyl OBD Column, 5 um, 19×250 mm; Mobile Phase A: Water (0.05% FA), Mobile Phase B: MeOH—HPLC; Flow rate: 25 mL/min; Gradient: 54% B to 70% B in 10 min; 254 nm) to afford N-[3-(2-chlorophenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (8.1 mg, 10%) as a white solid. LCMS (ESI, m/z): [M+H]+=312.1. 1H NMR (300 MHz, DMSO-d6): δ 11.79 (s, 1H), 10.64 (s, 1H), 7.98-7.91 (m, 1H), 7.87-7.84 (m, 1H), 7.60-7.57 (m, 3H), 7.49-7.30 (m, 2H), 2.10-2.00 (m, 1H), 0.83-0.79 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.83 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 2-(trifluoromethyl)phenylboronic acid (189.0 mg, 0.99 mmol), K2CO3 (343.8 mg, 2.49 mmol) and Pd(dppf)Cl2 (60.7 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (94/6, v/v) to afford 6-chloro-3-[2-(trifluoromethyl)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 84%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=427.1.
To a solution of 6-chloro-3-[2-(trifluoromethyl)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (250.0 mg, 0.59 mmol) in 1,4-dioxane (5.0 mL) was added cyclopropanecarboxamide (149.5 mg, 1.76 mmol), Cs2CO3 (572.4 mg, 1.76 mmol), BrettPhos (62.8 mg, 0.12 mmol) and Brettphos Pd G3 (53.1 mg, 0.06 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (84/16, v/v) to afford N-[3-[2-(trifluoromethyl)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (200.0 mg, 71%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=476.2.
The solution of N-[3-[2-(trifluoromethyl)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (200.0 mg, 0.42 mmol) in CH2Cl2/TFA (5.0 mL/5.0 mL) was stirred at room temperature for 1 h. The mixture was evaporated in vacuo. The residue was re-dissolved in ACN/H2O (5.0/1.0 mL). Then K2CO3 (581.2 mg, 4.21 mmol) was added to the mixture. The mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 48% B to 61% B in 7 min; 254 nm) to afford N-[3-[2-(trifluoromethyl)phenyl]-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (21.0 mg, 14%) as a white solid. LCMS (ESI, m/z): [M+H]+=346.2. 1H NMR (300 MHz, DMSO-d6): δ 11.70 (s, 1H), 10.64 (s, 1H), 7.90-7.84 (m, 2H), 7.73-7.70 (m, 2H), 7.60-7.56 (m, 2H), 7.37 (s, 1H), 2.05-1.95 (m, 1H), 0.85-0.75 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.83 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 2-(trifluoromethoxy)phenylboronic acid (204.9 mg, 0.99 mmol), K2CO3 (343.8 mg, 2.49 mmol) and Pd(dppf)Cl2 (60.7 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 3 h under N2. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 6-chloro-3-[2-(trifluoromethoxy)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (290.0 mg, 78%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=443.1.
To a solution of 6-chloro-3-[2-(trifluoromethoxy)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (240.0 mg, 0.54 mmol) in 1,4-dioxane (5.0 mL) was added cyclopropanecarboxamide (138.3 mg, 1.63 mmol), BrettPhos (58.2 mg, 0.11 mmol), Cs2CO3 (529.6 mg, 1.63 mmol) and BrettPhos Pd G3 (49.1 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 3 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford N-[3-[2-(trifluoromethoxy)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (155.0 mg, 58%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=492.2.
To a solution of N-[3-[2-(trifluoromethoxy)phenyl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (155.0 mg, 0.32 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the reaction mixture was evaporated in vacuo. The residue was dissolved with ACN/H2O (5.0/3.0 mL). Then K2CO3 (435.8 mg, 3.45 mmol) was added to the mixture. The mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 36% B to 56% B in 11 min; 254 nm) to afford N-[3-[2-(trifluoromethoxy)phenyl]-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (13.7 mg, 12%) as a white solid. LCMS (ESI, m/z): [M+H]+=362.1. 1H NMR (300 MHz, DMSO-d6): δ 11.79 (s, 1H), 10.66 (s, 1H), 7.99-7.92 (m, 2H), 7.75-7.71 (m, 1H), 7.60 (d, J=2.4 Hz, 1H), 7.51-7.42 (m, 3H), 2.05-1.95 (m, 1H), 0.83-0.79 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.83 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 2-cyanophenylboronic acid (146.2 mg, 1.00 mmol), K2CO3 (343.9 mg, 2.49 mmol) and Pd(dppf)Cl2 (60.7 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 3 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (65/35, v/v) to afford 2-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)benzonitrile (190.0 mg, 59%) as a white solid. LCMS (ESI, m/z): [M+H]+=384.1.
To a solution of 2-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)benzonitrile (140.0 mg, 0.37 mmol) in 1,4-dioxane (5.0 mL) was added cyclopropanecarboxamide (93.1 mg, 1.09 mmol), Cs2CO3 (356.4 mg, 1.09 mmol), Brettphos (39.14 mg, 0.07 mmol) and Brettphos Pd G3 (33.1 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (7/3, v/v) to afford N-[3-(2-cyanophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (150.0 mg, 95%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=433.2.
To a solution of N-[3-(2-cyanophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (150.0 mg, 0.35 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL). The resulting mixture stirred at room temperature for 1 h. The reaction mixture was evaporated in vacuo. The residue was re-dissolved in ACN/H2O (5.0/1.0 mL). Then K2CO3 (239.6 mg, 1.73 mmol) was added to the mixture. The mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 42% B in 11 min; 254 nm) to afford N-[3-(2-cyanophenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (24.9 mg, 18%) as a white solid. LCMS (ESI, m/z): [M+H]+=303.2. 1H NMR (300 MHz, DMSO-d6): δ 11.93 (s, 1H), 10.68 (s, 1H), 8.02-7.91 (m, 3H), 7.79-7.76 (m, 3H), 7.51-7.46 (m, 1H), 2.06-2.03 (m, 1H), 0.83-0.79 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.83 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (204.1 mg, 1.0 mmol), K2CO3 (343.9 mg, 2.49 mmol) and Pd(dppf)Cl2 (60.7 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 3 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/9, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridine (290.0 mg, 97%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=360.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridine (250.0 mg, 0.69 mmol) in 1,4-dioxane (5.0 mL) was added cyclopropanecarboxamide (177.3 mg, 2.08 mmol), Cs2CO3 (678.9 mg, 2.08 mmol), Brettphos (74.6 mg, 0.14 mmol) and Brettphos Pd G3 (62.9 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (14/86, v/v) to afford N-[3-(pyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (200.0 mg, 70%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=409.2.
To a solution of N-[3-(pyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (150.0 mg, 0.37 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL). The resulting mixture was stirred at room temperature for 1 h. The reaction mixture was evaporated in vacuo. The residue was re-dissolved in ACN/H2O (5.0/1.0 mL). Then K2CO3 (253.7 mg, 1.84 mmol) was added to the mixture. The mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH F-Phenyl OBD column, 19×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL/min; Gradient: 32% B to 51% B in 7 min; 254 nm) to afford N-[3-(pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (18.3 mg, 18%) as a white solid. LCMS (ESI, m/z): [M+H]+=279.2. 1H NMR (300 MHz, DMSO-d6): δ 11.82 (s, 1H), 10.66 (s, 1H), 8.97 (d, J=1.5 Hz, 1H), 8.44 (d, J=3.3 Hz, 1H), 8.30 (d, J=8.7 Hz, 1H), 8.13-8.11 (m, 1H), 7.97 (d, J=9.0 Hz, 1H), 7.91 (d, J=2.4 Hz, 1H), 7.46-7.42 (m, 1H), 2.19-1.90 (m, 1H), 0.84-0.80 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (280.0 mg, 0.77 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added pyridin-4-ylboronic acid (95.1 mg, 0.77 mmol), K2CO3 (320.9 mg, 2.32 mmol) and Pd(dppf)Cl2 (113.3 mg, 0.16 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (70/30, v/v) to afford 4-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridine (114.0 mg, 41%) as a white solid. LCMS (ESI, m/z): [M+H]+=360.1.
To a solution of 4-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridine (120.0 mg, 0.33 mmol) in dioxane (8.0 mL) was added cyclopropanecarboxamide (141.9 mg, 1.67 mmol), BrettPhos (71.6 mg, 0.13 mmol), Cs2CO3 (325.9 mg, 1.00 mmol) and BrettPhos Pd G3 (60.5 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (64/36, v/v) to afford N-[3-(pyridin-4-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (100.0 mg, 73%) as a white solid. LCMS (ESI, m/z): [M+H]+=409.2.
To a stirred mixture of N-[3-(pyridin-4-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (100.0 mg, 0.25 mmol) in DCM (3.0 mL) was added TFA (3.0 mL). The resulting mixture was stirred at room temperature for 3 h. The mixture was evaporated in vacuo. The residue was re-dissolved in ACN/H2O (5.0/1.0 mL). Then K2CO3 (182.7 mg, 1.32 mmol) was added to the mixture. The mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH F-Phenyl OBD column, 19×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL/min; Gradient: 30% B to 60% B in 7 min; 254 nm) to afford N-[3-(pyridin-4-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (12.8 mg, 10%) as a white solid. LCMS (ESI, m/z): [M+H]+=279.1. 1H NMR (400 MHz, DMSO-d6): δ 11.95 (s, 1H), 10.65 (s, 1H), 8.53 (d, J=4.8 Hz, 2H), 8.39 (d, J=8.8 Hz, 1H), 8.08 (s, 1H), 7.99 (d, J=8.8 Hz, 1H), 7.74 (d, J=4.8 Hz, 2H), 2.05-1.97 (m, 1H), 0.83-0.79 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (280.0 mg, 0.77 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 2-hydroxyphenylboronic acid (106.8 mg, 0.77 mmol), K2CO3 (320.9 mg, 2.32 mmol) and Pd(dppf)Cl2 (113.3 mg, 0.16 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (89/11, v/v) to afford 2-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)phenol (110.0 mg, 38%) as a white solid. LCMS (ESI, m/z): [M+H]+=375.1.
To a solution of 2-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)phenol (110.0 mg, 0.29 mmol) in dioxane (8.0 mL) was added cyclopropanecarboxamide (124.8 mg, 1.47 mmol), BrettPhos (63.0 mg, 0.12 mmol), Cs2CO3 (286.8 mg, 0.88 mmol) and BrettPhos Pd G3 (53.2 mg, 0.06 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (63/37, v/v) to afford N-[3-(2-hydroxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (52.0 mg, 42%) as a white solid. LCMS (ESI, m/z): [M+H]+=424.2.
To a stirred mixture of N-[3-(2-hydroxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (110.0 mg, 0.26 mmol) in DCM (3.0 mL) was added TFA (3.0 mL). The resulting mixture was stirred at room temperature for 3 h. The mixture was evaporated in vacuo. The residue was re-dissolved in ACN/H2O (5.0/1.0 mL). Then K2CO3 (107.7 mg, 0.78 mmol) was added to the mixture. The mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 24% B to 35% B in 12 min; 254 nm) to afford N-[3-(2-hydroxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (16.9 mg, 20%) as a white solid. LCMS (ESI, m/z): [M+H]+=294.2. 1H NMR (400 MHz, DMSO-d6): δ 11.47 (s, 1H), 10.55 (s, 1H), 9.48 (s, 1H), 8.05 (d, J=8.4 Hz, 1H), 7.87 (d, J=8.4 Hz, 1H), 7.61 (s, 1H), 7.50 (d, J=8.0 Hz, 1H), 7.08-7.05 (m, 1H), 6.94 (d, J=7.6 Hz, 1H), 6.89-6.86 (m, 1H), 2.05-1.97 (m, 1H), 0.82-0.78 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.83 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 3-hydroxyphenylboronic acid (137.3 mg, 0.99 mmol), K2CO3 (343.9 mg, 2.49 mmol) and Pd(dppf)Cl2 (60.7 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (88/12, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)phenol (230.0 mg, 73%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=375.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)phenol (200.0 mg, 0.53 mmol) in 1,4-dioxane (5.0 mL) was added cyclopropanecarboxamide (136.2 mg, 1.60 mmol), Cs2CO3 (521.4 mg, 1.60 mmol), Brettphos (57.3 mg, 0.11 mmol) and Brettphos Pd G3 (48.4 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (7/3, v/v) to afford N-[3-(3-hydroxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (220.0 mg, 97%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=424.2.
The solution of N-[3-(3-hydroxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (170.0 mg, 0.40 mmol) in CH2Cl2/TFA (5.0 mL/5.0 mL) was stirred at room temperature for 1 h. The mixture was evaporated in vacuo. The residue was re-dissolved in ACN/H2O (5.0/1.0 mL). Then K2CO3 (554.7 mg, 4.01 mmol) was added to the mixture. The mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 29% B to 32% B in 8 min; 254 nm) to afford N-[3-(3-hydroxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (34.0 mg, 28%) as a white solid. LCMS (ESI, m/z): [M+H]+=294.2. 1H NMR (300 MHz, DMSO-d6): δ 11.62 (s, 1H), 10.62 (s, 1H), 9.38 (s, 1H), 8.18 (d, J=8.7 Hz, 1H), 7.94 (d, J=8.7 Hz, 1H), 7.66 (d, J=2.7 Hz, 1H), 7.24-7.19 (m, 1H), 7.14-7.11 (m, 2H), 6.67-6.63 (m, 1H), 2.08-2.03 (m, 1H), 0.83-0.79 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in 1,4-dioxane/H2O (10.0/2.0 mL) was added 2-methoxypyridin-3-ylboronic acid (211.4 mg, 1.38 mmol), K2CO3 (57.3 mg, 0.42 mmol) and Pd(dppf)Cl2 (112.9 mg, 0.14 mmol). The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/EtOAc (80/20, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-methoxypyridine (250.0 mg, 46%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=390.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-methoxypyridine (235.0 mg, 0.60 mmol) in 1,4-dioxane (8.0 mL) was added cyclopropanecarboxamide (205.2 mg, 2.41 mmol), BrettPhos (64.7 mg, 0.12 mmol), Cs2CO3 (589.0 mg, 1.81 mmol) and Brettphos Pd G3 (54.6 mg, 0.06 mmol). The resulting mixture was stirred at 100° C. for 2 h under N2. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with ether/ethyl acetate (75/25, v/v) to afford N-(3-(2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (250.0 mg, 94%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=439.2.
To a solution of N-[3-(2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (250.0 mg, 0.57 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL). The resulting mixture was stirred at room temperature for 1 h. The mixture was concentrated under vacuum. The residue was re-dissolved in ACN (6.0 mL) and H2O (0.5 mL). Then K2CO3 (393.9 mg, 2.85 mmol) was added to the mixture. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with ether/ethyl acetate (82/18, v/v) and then purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18 Column, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 56% B in 10 min; 254 nm) to afford N-[3-(2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (51.0 mg, 29%) as a white solid. LCMS (ESI, m/z): [M+H]+=309.2. 1H NMR (300 MHz, DMSO-d6): δ 11.68 (s, 1H), 10.59 (s, 1H), 8.11-8.07 (m, 2H), 8.01-7.91 (m, 2H), 7.72 (d, J=2.7 Hz, 1H), 7.10-7.06 (m, 1H), 3.95 (s, 3H), 2.08-2.01 (m, 1H), 0.87-0.79 (m, 4H).
To a mixture of 2,4-dinitroaniline (10.0 g, 54.61 mmol) and KOH (9.2 g, 163.80 mmol) in MeOH (1.0 L) was added dropwise a solution of potassium hydroxide (64.9 g, 546.12 mmol) in H2O (1.0 L) 50° C. under N2. The resulting mixture was stirred at 50° C. for 3 h. After the reaction was completed, the reaction was cooled to room temperature and then filtered. The precipitated solid was collected and dried to afford 5-bromo-4-methoxy-2,1,3-benzoxadiazol-1-ium-1-olate (2.1 g, 16%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=244.9.
To a solution of 5-bromo-4-methoxy-2,1,3-benzoxadiazol-1-ium-1-olate (1.2 g, 4.90 mmol) in EtOAc (30.0 mL) was added Pd/C (0.1 g, dry). The resulting mixture was stirred at room temperature for 16 h under H2. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure to afford 4-bromo-3-methoxybenzene-1,2-diamine (623.0 mg, crude) as a brown solid. LCMS (ESI, m/z): [M+H]+=217.0.
The solution of 4-bromo-3-methoxybenzene-1,2-diamine (623.0 mg, 2.87 mmol) in HCOOH (10.0 mL) was stirred at 100° C. for 2 h. After the reaction was completed, the reaction was diluted with H2O. The pH value of the mixture was adjusted to 8 with NaOH (aq.) and then extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) to afford 5-bromo-4-methoxy-3H-1,3-benzodiazole (495.0 mg, 75%) as a red solid. LCMS (ESI, m/z): [M+H]+=227.0.
To a solution of 5-bromo-4-methoxy-3H-1,3-benzodiazole (710.0 mg, 3.13 mmol) in THE (20.0 mL) was added NaH (375.2 mg, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. Then SEM-Cl (782.0 mg, 4.69 mmol) was added dropwise to the mixture at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h under N2. After the reaction was completed, the reaction was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 6-bromo-7-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (790.0 mg, crude) as a black oil. LCMS (ESI, m/z): [M+H]+=357.1.
To a solution of 6-bromo-7-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (740.0 mg, 2.07 mmol) in dioxane (20.0 mL) was added bis(pinacolato)diboron (1577.7 mg, 6.21 mmol), KOAc (609.7 mg, 6.21 mmol) and Pd(dppf)Cl2 (169.1 mg, 0.21 mmol) at room temperature under N2. The resulting mixture was stirred at 85° C. for 16 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (6/1, v/v) to afford 7-methoxy-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (270.0 mg, 32%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=405.2.
To a solution of 7-methoxy-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (405.0 mg, 1.00 mmol) in dioxane/H2O (10.0 mL/2.0 mL) was added 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (301.9 mg, 0.84 mmol), K2CO3 (346.0 mg, 2.50 mmol) and Pd(dppf)Cl2 (68.2 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (140.0 mg, 30%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=559.2.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (122.0 mg, 0.22 mmol) in 1,4-dioxane (6.0 mL) was added cyclopropanecarboxamide (92.8 mg, 1.09 mmol), BrettPhos (23.4 mg, 0.04 mmol), Cs2CO3 (213.2 mg, 0.65 mmol) and BrettPhos Pd G3 (19.7 mg, 0.02 mmol) at room temperature under N2. The resulting mixture was irradiated with microwave radiation at 120° C. for 1.5 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford N-[3-(4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (109.0 mg, 82%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=608.3.
To a solution of N-[3-(4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (109.0 mg, 0.18 mmol) in DMF (5.0 mL) was added ethane-1,2-diamine (53.8 mg, 0.90 mmol) and TBAF (0.5 mL, 0.01 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions Column: (Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 16% B to 38% B in 6 min; 254 nm) to afford N-[3-(4-methoxy-1H-1,3-benzodiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (9.1 mg, 15%) as a white solid. LCMS (ESI, m/z): [M+H]+=348.2. 1H NMR (300 MHz, DMSO-d6): δ 12.49 (s, 1H), 11.47 (s, 1H), 10.58 (s, 1H), 8.21-8.18 (m, 1H), 7.97-7.87 (m, 2H), 7.47 (s, 1H), 7.36-7.33 (m, 1H), 7.26-7.24 (m, 1H), 4.22 (s, 2H), 3.66 (s, 1H), 2.10-2.01 (m, 1H), 0.83-0.79 (m, 4H).
To a mixture of 5-bromo-4-methoxy-1H-indazole (1.0 g, 4.40 mmol) in THF (10.0 mL) was added NaH (0.3 g, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. Then SEM-Cl (1.1 g, 6.62 mmol) was added dropwise to the mixture at 0° C. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the reaction mixture was quenched with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 5-bromo-4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (2.5 g, crude) as a yellow oil. LCMS (ESI, m/z): [M+H]+=357.1.
To a mixture of 5-bromo-4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (2.4 g, 6.71 mmol) and 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (5.1 g, 20.16 mmol) in dioxane (30.0 mL) was added Pd(dppf)Cl2 (0.5 g, 0.67 mmol) and KOAc (1.9 g, 20.17 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 4-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy] methyl]indazole (750.0 mg, 27%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=405.2.
To a mixture of 4-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (660.0 mg, 1.63 mmol) and 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (400.0 mg, 1.11 mmol) in dioxane/H2O (10.0/1.0 mL) was added Pd(dppf)Cl2 (90.0 mg, 0.11 mmol) and K2CO3 (460.0 mg, 3.33 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl] indazole (330.0 mg, 36%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=559.2.
To a mixture of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (300.0 mg, 0.53 mmol) and cyclopropanecarboxamide (182.6 mg, 2.14 mmol) in dioxane (5.0 mL) was added BrettPhos Pd G3 (48.6 mg, 0.05 mmol), BrettPhos (57.5 mg, 0.10 mmol) and Cs2CO3 (699.1 mg, 2.14 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford N-[3-(4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (220.0 mg, 67%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=608.3.
To a mixture of N-[3-(4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (190.0 mg, 0.31 mmol) in DCM (3.0 mL) was added TFA (1.0 mL). The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in CH3CN/H2O (5.0/1.0 mL). Then K2CO3 (500.0 mg. 3.62 mmol) was added to the mixture. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 28% B to 48% B in 7 min; 254 nm) to afford N-[3-(4-methoxy-1H-indazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (17.1 mg, 15%) as a white solid. LCMS (ESI, m/z): [M+H]+=348.1. 1H NMR (300 MHz, DMSO-d6): δ 13.22 (s, 1H), 11.35 (s, 1H), 10.45 (s, 1H), 8.43 (s, 1H), 7.88-7.74 (m, 3H), 7.26 (d, J=8.7 Hz, 1H), 6.77 (d, J=2.1 Hz, 1H), 4.25 (s, 3H), 2.11-2.02 (m, 1H), 0.87-0.79 (m, 4H).
To a solution of 6-bromo-5-methoxy-1H-indole (1.0 g, 4.42 mmol) in 1,4-dioxane (8.0 mL) was added 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (3.4 g, 13.27 mmol), KOAc (1.3 g, 13.22 mmol) and Pd(dppf)Cl2 (0.4 g, 0.44 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to afford 5-methoxy-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole (900.0 mg, 74%) as a white solid. LCMS (ESI, m/z): [M+H]+=274.2.
To a solution of 5-methoxy-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole (350.0 mg, 1.28 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (579.4 mg, 1.28 mmol), K2CO3 (531.3 mg, 3.84 mmol) and Pd(dppf)Cl2 (104.6 mg, 0.13 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (89/11, v/v) to afford 6-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-methoxy-1H-indole (200.0 mg, 36%) as a white solid. LCMS (ESI, m/z): [M+H]+=428.1.
To a solution of 6-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-methoxy-1H-indole (180.0 mg, 0.42 mmol) in dioxane (3.0 mL) was added cyclopropanecarboxamide (143.1 mg, 1.68 mmol), BrettPhos (90.3 mg, 0.17 mmol), Cs2CO3 (411.1 mg, 1.26 mmol) and BrettPhos Pd G3 (76.2 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (81/19, v/v) to afford N-[3-(5-methoxy-1H-indol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (159.0 mg, 79%) as a white solid. LCMS (ESI, m/z): [M+H]+=477.2.
To a solution of N-[3-(5-methoxy-1H-indol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (150.0 mg, 0.31 mmol) in DMF (2.0 mL) was added EDA (94.5 mg, 1.57 mmol) and TBAF (246.8 mg, 0.94 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 27% B to 54% B in 8 min; 254 nm) to afford N-[3-(5-methoxy-1H-indol-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (14.9 mg, 14%) as a white solid. LCMS (ESI, m/z): [M+H]+=347.2. 1H NMR (400 MHz, DMSO-d6): δ 11.43 (s, 1H), 10.85 (s, 1H), 10.56 (s, 1H), 7.96 (d, J=8.8 Hz, 1H), 7.88 (d, J=8.8 Hz, 1H), 7.50 (s, 2H), 7.28 (s, 1H), 7.18 (s, 1H), 6.37 (s, 1H), 3.78 (s, 3H), 2.07-2.03 (m, 1H), 0.83-0.77 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 1H-indol-5-ylboronic acid (222.5 mg, 1.38 mmol), K2CO3 (573.1 mg, 4.15 mmol) and Pd(dppf)Cl2 (225.7 mg, 0.27 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (84/16, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-1H-indole (220.0 mg, 40%) as a white solid. LCMS (ESI, m/z): [M+H]+=398.1.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-1H-indole (200.0 mg, 0.50 mmol) in dioxane (3.0 mL) was added cyclopropanecarboxamide (128.3 mg, 1.51 mmol), BrettPhos (107.9 mg, 0.20 mmol), Cs2CO3 (491.2 mg, 1.51 mmol) and BrettPhos Pd G3 (91.1 mg, 0.10 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (79/21, v/v) to afford N-[3-(1H-indol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (150.0 mg, 66%) as a white solid. LCMS (ESI, m/z): [M+H]+=447.2.
To a solution of N-[3-(1H-indol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (100.0 mg, 0.22 mmol) in DMF (2.0 mL) was added EDA (40.4 mg, 0.67 mmol) and TBAF (292.7 mg, 1.12 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 27% B to 49% B in 9 min; 254 nm) to afford N-[3-(1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (26.7 mg, 38%) as a white solid. LCMS (ESI, m/z): [M+H]+=317.2. 1H NMR (400 MHz, DMSO-d6): δ 11.45 (s, 1H), 11.04 (s, 1H), 10.58 (s, 1H), 8.22 (d, J=8.4 Hz, 1H), 7.92 (d, J=8.8 Hz, 1H), 7.83 (s, 1H), 7.58 (d, J=1.6 Hz, 1H), 7.46-7.41 (m, 2H), 7.33 (d, J=2.4 Hz, 1H), 6.46 (s, 1H), 2.07-2.03 (m, 1H), 0.83-0.79 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-1,3-benzodiazole (404.9 mg, 1.66 mmol), K2CO3 (573.1 mg, 4.15 mmol) and Pd(dppf)Cl2 (112.9 mg, 0.14 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with ethyl acetate/MeOH (10/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-1H-1,3-benzodiazole (280.0 mg, 50%) as a white solid. LCMS (ESI, m/z): [M+H]+=399.1.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-1H-1,3-benzodiazole (280.0 mg, 0.70 mmol) in 1,4-dioxane (10.0 mL) was added cyclopropanecarboxamide (1194.5 mg, 14.04 mmol), BrettPhos (75.34 mg, 0.14 mmol), Cs2CO3 (686.0 mg, 2.10 mmol) and BrettPhos Pd G3 (63.6 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was irradiated with microwave radiation at 120° C. for 1.5 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with ethyl acetate/MeOH (10/1, v/v) to afford N-[3-(1H-1,3-benzodiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (250.0 mg, 79%) as a white solid. LCMS (ESI, m/z): [M+H]+=448.2.
To a solution of N-[3-(1H-1,3-benzodiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (400.0 mg, 0.89 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The reaction mixture was evaporated in vacuo. The residue was re-dissolved in ACN/H2O (10.0/2.0 mL). Then K2CO3 (1235.0 mg, 8.94 mmol) was added to the mixture. The mixture was stirred at room temperature for another 16 h. After the reaction was completed, the mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 15% B to 45% B in 10 min; 254 nm) to afford N-[3-(1H-1,3-benzodiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (9.0 mg, 3%) as a white solid. LCMS (ESI, m/z): [M+H]+=318.1. 1H NMR (300 MHz, DMSO-d6): δ 12.41-12.35 (m, 1H), 11.59-11.55 (m, 1H), 10.60 (s, 1H), 8.25-8.19 (m, 2H), 7.97-7.54 (m, 5H), 2.08-2.03 (m, 1H), 0.84-0.79 (m, 4H).
To a solution of 3-bromo-6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in dioxane/H2O (8.0/2.0 mL) was added 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole (336.0 mg, 1.38 mmol), K2CO3 (573.1 mg, 4.15 mmol) and Pd(dppf)Cl2 (101.1 mg, 0.14 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 6-chloro-3-(1H-indol-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (160.0 mg, 29%) as a white solid. LCMS (ESI, m/z): [M+H]+=398.1.
To a solution of 6-chloro-3-(1H-indol-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (160.0 mg, 0.40 mmol) in 1,4-dioxane (10.0 mL) was added cyclopropanecarboxamide (684.3 mg, 8.04 mmol), Cs2CO3 (393.0 mg, 1.21 mmol) and Brettphos (43.2 mg, 0.08 mmol), BrettPhos Pd G3 (36.4 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (6/1, v/v) to afford N-(3-(1H-indol-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (130.0 mg, 72%) as a white solid. LCMS (ESI, m/z): [M+H]+=447.2.
To a solution of N-(3-(1H-indol-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (100.0 mg, 0.22 mmol) in DMF (5.0 mL) was added TBAF (0.6 mL) and EDA (67.2 mg, 1.12 mmol). The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with 5-100% CH3CN in H2O and then purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 30% B to 57% B in 8 min; 220 nm) to afford N-(3-(1H-indol-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (1.4 mg, 1%) as a white solid. LCMS (ESI, m/z): [M+H]+=317.2. 1H NMR (300 MHz, DMSO-d6): δ 11.53 (s, 1H), 11.01 (s, 1H), 10.61 (s, 1H), 8.23 (d, J=9.0 Hz, 1H), 7.95 (d, J=8.4 Hz, 1H), 7.68-7.63 (m, 2H), 7.59 (d, J=8.1 Hz, 1H), 7.36-7.33 (m, 2H), 6.43 (s, 1H), 2.08-1.98 (m, 1H), 0.84-0.79 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in dioxane/H2O (8.0/2.0 mL) was added tert-butyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole-1-carboxylate (475.8 mg, 1.38 mmol), K2CO3 (573.1 mg, 4.15 mmol) and Pd(dppf)Cl2 (101.1 mg, 0.14 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford tert-butyl 5-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-1H-indazole-1-carboxylate (140.0 mg, 20%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=499.2.
To a solution of tert-butyl 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)indazole-1-carboxylate (110.0 mg, 0.22 mmol) in 1,4-dioxane (10.0 mL) was added cyclopropanecarboxamide (375.2 mg, 4.41 mmol), Cs2CO3 (215.4 mg, 0.66 mmol), Brettphos (23.7 mg, 0.04 mmol) and BrettPhos Pd G3 (20.0 mg, 0.02 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford tert-butyl 5-(6-(cyclopropanecarboxamido)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-1H-indazole-1-carboxylate (110.0 mg, 91%) as a white solid. LCMS (ESI, m/z): [M+H]+=548.3.
To a solution of tert-butyl 5-(6-(cyclopropanecarboxamido)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-1H-indazole-1-carboxylate (110.0 mg, 0.20 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL). The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in CH3CN/H2O (20.0/20.0 mL). Then K2CO3 (277.6 mg, 2.00 mmol) was added to the mixture. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/2, v/v) and then purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 39% B in 8 min; 254 nm) to afford N-[3-(1H-indazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (9.3 mg, 14%) as a white solid. LCMS (ESI, m/z): [M+H]+=318.2. 1H NMR (400 MHz, DMSO-d6): δ 13.02 (s, 1H), 11.58 (s, 1H), 10.62 (s, 1H), 8.29 (d, J=8.8 Hz, 1H), 8.09-8.05 (m, 2H), 7.96 (d, J=8.8 Hz, 1H), 7.74-7.71 (m, 2H), 7.60 (d, J=8.8 Hz, 1H), 2.07-1.99 (m, 1H), 0.85-0.80 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in 1,4-dioxane/H2O (8.0/2.0 mL) was added 2-fluoro-6-methoxyphenylboronic acid (281.9 mg, 1.66 mmol), K2CO3 (382.1 mg, 2.76 mmol) and Pd(PPh3)4 (159.7 mg, 0.14 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 6-chloro-3-(2-fluoro-6-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (220.0 mg, 39%) as a brown oil. LCMS (ESI, m/z): [M+H]+=407.1.
To a solution of 6-chloro-3-(2-fluoro-6-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (250.0 mg, 0.61 mmol) in 1,4-dioxane (6.0 mL) was added (1R,2R)-2-fluorocyclopropane-1-carboxamide (316.7 mg, 3.07 mmol), BrettPhos (66.0 mg, 0.12 mmol), Cs2CO3 (600.5 mg, 1.84 mmol) and BrettPhos Pd G3 (55.7 mg, 0.06 mmol) at room temperature. The resulting mixture was irradiated with microwave radiation at 120° C. for 1.5 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with ether/ethyl acetate (3/2, v/v) to afford (1R,2R)-2-fluoro-N-[3-(2-fluoro-6-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (146.0 mg, 50%) as a brown yellow oil. LCMS (ESI, m/z): [M+H]+=474.2.
To a solution of (1R,2R)-2-fluoro-N-[3-(2-fluoro-6-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (146.0 mg, 0.31 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The mixture was evaporated under reduced pressure. The residue was re-dissolved in ACN/H2O (10.0/2.0 mL). Then K2CO3 (426.1 mg, 3.08 mmol) was added to the mixture. The mixture was stirred at room temperature for another 16 h. After the reaction was completed, the reaction mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 31% B to 61% B in 10 min; 254 nm) to afford (1R,2R)-2-fluoro-N-[3-(2-fluoro-6-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (10.8 mg, 10%) as a white solid. LCMS (ESI, m/z): [M+H]+=344.1. 1H NMR (400 MHz, DMSO-d6): δ 11.67 (s, 1H), 10.63 (s, 1H), 7.88-7.86 (m, 1H), 7.70-7.68 (m, 1H), 7.46 (s, 1H), 7.37-7.31 (m, 1H), 6.99-6.90 (m, 2H), 5.03-4.82 (m, 1H), 3.78 (s, 3H), 2.27-2.20 (m, 1H), 1.71-1.63 (m, 1H), 1.20-1.05 (m, 1H).
To a solution of 2,2-difluorocyclopropane-1-carboxylic acid (800.0 mg, 6.55 mmol) in DCM (10.0 mL) was added DMF (0.1 mL) and oxalyl chloride (998.1 mg, 7.86 mmol). The resulting mixture was stirred at room temperature for 1 h. Then NH3 (g)/MeOH (10.0 mL, 7 mol/L) was added to the mixture. The mixture was stirred at room temperature for another 0.5 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to afford 2,2-difluorocyclopropane-1-carboxamide (790.0 mg, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=122.0.
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in 1,4-dioxane/H2O (20.0/4.0 mL) was added 2-methoxyphenylboronic acid (252.1 mg, 1.66 mmol), K2CO3 (573.1 mg, 4.15 mmol) and Pd(dppf)Cl2 (112.9 mg, 0.14 mmol). The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 6-chloro-3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (230.0 mg, 42%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=389.1.
To a solution of 6-chloro-3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (170.0 mg, 0.44 mmol) in 1,4-dioxane (6.0 mL) was added 2,2-difluorocyclopropane-1-carboxamide (264.6 mg, 2.19 mmol), Cs2CO3 (427.2 mg, 1.31 mmol), BrettPhos (46.9 mg, 0.09 mmol) and BrettPhos Pd G3 (396.2 mg, 0.48 mmol). The resulting mixture was irradiated with microwave radiation at 120° C. for 1.5 h under N2. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 2,2-difluoro-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (120.0 mg, 57%) as a brown yellow oil. LCMS (ESI, m/z): [M+H]+=474.2.
To a solution of 2,2-difluoro-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (120.0 mg, 0.25 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL). The resulting mixture was stirred at room temperature for 3 h. The reaction mixture was evaporated under reduced pressure. The residue was re-dissolved in ACN/H2O (10.0/2.0 mL). Then K2CO3 (350.2 mg, 2.53 mmol) was added to the mixture. The mixture was stirred at room temperature for another 16 h. After the reaction was completed, the mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep Phenyl OBD Column, 19×150 mm, 5 um, 13 nm; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 32% B to 62% B in 10 min; 254 nm) to afford 2,2-difluoro-N-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (14.7 mg, 16%) as a white solid. LCMS (ESI, m/z): [M+H]+=344.1. 1H NMR (400 MHz, DMSO-d6): δ 11.64 (s, 1H), 10.82 (s, 1H), 8.02 (d, J=8.4 Hz, 1H), 7.88 (d, J=8.4 Hz, 1H), 7.60 (d, J=2.0 Hz, 1H), 7.54 (d, J=7.2 Hz, 1H), 7.30-7.26 (m, 1H), 7.12 (d, J=8.0 Hz, 1H), 7.05-7.01 (m, 1H), 3.82 (s, 3H), 3.06-2.98 (m, 1H), 2.09-1.95 (m, 2H).
To a solution of tert-butyl 5-bromo-2,3-dihydroindole-1-carboxylate (1.0 g, 3.35 mmol) in 1,4-dioxane (10.0 mL) was added bis(pinacolato)diboron (2.6 g, 10.06 mmol), KOAc (987.4 mg, 10.06 mmol) and Pd(dppf)Cl2 (112.6 mg, 0.15 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (89/11, v/v) to afford tert-butyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydroindole-1-carboxylate (1.0 g, 73%) as a red solid. LCMS (ESI, m/z): [M+H]+=346.2.
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (360.0 mg, 0.99 mmol) in 1,4-dioxane/H2O (5.0 mL/1.0 mL) was added tert-butyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydroindole-1-carboxylate (412.3 mg, 1.19 mmol), K2CO3 (412.6 mg, 2.99 mmol), and Pd(dppf)Cl2 (72.8 mg, 0.10 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (68/32, v/v) to afford tert-butyl 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2,3-dihydroindole-1-carboxylate (260.0 mg, 52%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=500.2.
To a solution of tert-butyl 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2,3-dihydroindole-1-carboxylate (200.0 mg, 0.40 mmol) in 1,4-dioxane (5.0 mL) was added cyclopropanecarboxamide (40.8 mg, 0.48 mmol), Cs2CO3 (390.9 mg, 1.20 mmol), BrettPhos (42.9 mg, 0.08 mmol) and Brettphos Pd G3 (36.2 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (52/48, v/v) to afford tert-butyl 5-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2,3-dihydroindole-1-carboxylate (200.0 mg, 91%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=549.2.
To a solution of tert-butyl 5-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2,3-dihydroindole-1-carboxylate (200.0 mg, 0.36 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL). The mixture was stirred at room temperature for 1 h. The reaction mixture was evaporated in vacuo. The residue was re-dissolved in CH3CN/NH3·H2O (5.0/5.0 mL). The mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 18% B to 48% B in 10 min; 254 nm) to afford N-[3-(2,3-dihydro-1H-indol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (5.2 mg, 4%) as a white solid. LCMS (ESI, m/z): [M+H]+=319.2. 1H NMR (300 MHz, DMSO-d6): δ 11.38 (s, 1H), 10.57 (s, 1H), 8.14 (d, J=8.7 Hz, 1H), 7.89 (d, J=8.7 Hz, 1H), 7.47 (d, J=2.4 Hz, 1H), 7.37 (s, 1H), 7.25 (d, J=7.8 Hz, 1H), 6.57 (d, J=8.1 Hz, 1H), 5.47 (s, 1H), 3.47-3.42 (m, 2H), 3.00-2.97 (m, 2H), 2.08-1.92 (m, 1H), 0.83-0.78 (m, 4H).
A mixture of tert-butyl 6-bromo-2,3-dihydroindole-1-carboxylate (1.0 g, 3.35 mmol), bis(pinacolato)diboron (1.3 g, 5.03 mmol), KOAc (1.0 g, 10.06 mmol) and Pd(dppf)Cl2 (0.3 g, 0.34 mmol) in dioxane (25.0 mL) was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (89/11, v/v) to afford tert-butyl 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydroindole-1-carboxylate (1.1 g, 95%) as a white solid. LCMS (ESI, m/z): [M+H]+=346.2.
To a solution of tert-butyl 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydroindole-1-carboxylate (1.1 g, 3.19 mmol) in dioxane (20.0 mL) and H2O (4.0 mL) was added 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (1.4 g, 3.87 mmol), K2CO3 (1.3 g, 9.42 mmol) and Pd(dppf)Cl2 (0.2 g, 0.27 mmol). The mixture was stirred at 100° C. for 3 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (92/8, v/v) to afford tert-butyl 6-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2,3-dihydroindole-1-carboxylate (919.0 mg, 57%) as a white solid. LCMS (ESI, m/z): [M+H]+=500.2.
A mixture of tert-butyl 6-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2,3-dihydroindole-1-carboxylate (500.0 mg, 1.00 mmol), cyclopropanecarboxamide (255.3 mg, 3.00 mmol), Cs2CO3 (977.2 mg, 3.00 mmol), BrettPhos (107.3 mg, 0.20 mmol) and BrettPhos Pd G3 (90.6 mg, 0.10 mmol) in dioxane (10.0 mL) was stirred at 100° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (71/29, v/v) to afford tert-butyl 6-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2,3-dihydroindole-1-carboxylate (437.4 mg, 79%) as a white solid. LCMS (ESI, m/z): [M+H]+=549.3.
To a solution of tert-butyl 6-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2,3-dihydroindole-1-carboxylate (200.0 mg, 0.36 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL). The mixture was stirred at room temperature for 2 h. The mixture was evaporated in vacuo. The residue was re-dissolved in CH3CN (5.0 mL). Then NH3·H2O (5.0 mL) was add to the mixture. The mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (94/6, v/v) and then purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient 28% B to 49% B in 7 min; 254 nm; RT1:6.27 min) to afford N-[3-(2,3-dihydro-1H-indol-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (13.1 mg, 11%) as a white solid. LCMS (ESI, m/z): [M+H]+=319.2. 1H NMR (400 MHz, CD3OD): δ 8.21-8.19 (m, 1H), 7.81 (d, J=8.8 Hz, 1H), 7.43 (s, 1H), 7.17 (d, J=7.2 Hz, 1H), 7.03-7.00 (m, 2H), 3.55-3.49 (m, 2H), 3.06-3.02 (m, 2H), 1.95-1.85 (m, 1H), 1.03-0.99 (m, 2H), 0.96-0.88 (m, 2H).
To a solution of 6-chloro-3-(2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (240.0 mg, 0.61 mmol) in dioxane (10.0 mL) were added Cs2CO3 (601.5 mg, 1.84 mmol), (1R,2R)-2-fluorocyclopropane-1-carboxamide (317.2 mg, 3.07 mmol), BrettPhos (132.1 mg, 0.24 mmol) and Brettphos Pd G3 (111.5 mg, 0.12 mmol) under N2. The mixture was irradiated with microwave radiation at 100° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with ether/ethyl acetate (88/12, v/v) to afford (1R,2R)-2-fluoro-N-(3-(2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (230.0 mg, 81%) as a white solid. LCMS (ESI, m/z): [M+H]+=457.2.
To a solution of (1R,2R)-2-fluoro-N-[3-(2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (230.0 mg, 0.50 mmol) in CH2Cl2 (15.0 mL) was added TFA (15.0 mL). The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under vacuum. The residue was re-dissolved in CH3CN (15.0 mL). Then NH3H2O (30.0 mL) was added to the mixture. The mixture was stirred at room temperature for another 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) and then purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 49% B in 8 min; 220 nm) to afford (1R,2R)-2-fluoro-N-(3-(2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (55.5 mg, 33%) as a white solid. LCMS (ESI, m/z): [M+H]+=327.2 1H NMR (300 MHz, DMSO-d6): δ 11.70 (s, 1H), 10.64 (s, 1H), 8.13-8.07 (m, 2H), 8.01-7.90 (m, 2H), 7.72 (d, J=2.7 Hz, 1H), 7.10-7.06 (m, 1H), 5.05-4.80 (m, 1H), 3.95 (s, 3H), 2.26-2.21 (m, 1H), 1.71-1.61 (m, 1H), 1.19-1.08 (m, 1H).
A mixture of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (3.0 g, 8.29 mmol), 2-methoxypyridin-3-ylboronic acid (735.6 mg, 4.81 mmol), K2CO3 (2.0 g, 14.43 mmol) and Pd(dppf)Cl2 (606.8 mg, 0.82 mmol) in dioxane (20.0 mL)/H2O (2.0 mL) was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (7/3, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-methoxypyridine (1400.0 mg, 43%) as a yellow oil. LCMS (ESI, m/z): [M+H]=390.0.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-methoxypyridine (400.0 mg, 1.02 mmol) in dioxane (8.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (634.5 mg, 6.15 mmol), BrettPhos (220.2 mg, 0.41 mmol), Cs2CO3 (1002.6 mg, 3.07 mmol) and Brettphos Pd G3 (185.9 mg, 0.20 mmol). The final reaction mixture was irradiated with microwave radiation at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (7/3, v/v) to afford (1S,2S)-2-fluoro-N-[3-(2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (125.0 mg, 26%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=457.0.
A mixture of (1S,2S)-2-fluoro-N-[3-(2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (100.0 mg, 0.21 mmol) and TFA (5.0 mL) in CH2Cl2 (5.0 mL) was stirred at room temperature for 1 h. The resulting mixture was concentrated under vacuum. The residue was dissolved in CH3CN (5.0 mL). Then NH3H2O (5.0 mL) was added to the mixture. The resulting mixture was stirred at room temperature for additional 1 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 46% B in 7 min; 220 nm) to afford (1S,2S)-2-fluoro-N-[3-(2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (18.8 mg, 26%) as a white solid. LCMS (ESI, m/z): [M+H]+=327.2. 1H NMR (300 MHz, DMSO-d6): δ 11.72 (s, 1H), 10.65 (s, 1H), 8.14-8.08 (m, 2H), 8.02-7.91 (m, 2H), 7.74 (d, J=2.4 Hz, 1H), 7.11-7.07 (m, 1H), 5.05-4.81 (m, 1H), 3.96 (s, 3H), 2.27-2.22 (m, 1H), 1.71-1.62 (m, 1H), 1.20-1.13 (m, 1H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in 1,4-dioxane/H2O (15.0/3.0 mL) was added 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-1,3-benzodiazole (674.8 mg, 2.77 mmol), K2CO3 ((573.1 mg, 4.15 mmol) and Pd(dppf)Cl2 (112.9 mg, 0.14 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (12/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-1H-1,3-benzodiazole (177.0 mg, 32%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=399.1.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-1H-1,3-benzodiazole (297.0 mg, 0.74 mmol) in THF (20.0 mL) was added NaH (186.2 mg, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. Then SEM-Cl (186.2 mg, 1.12 mmol) was added dropwise to the mixture at 0° C. under N2. The resulting mixture was stirred at 0° C. for another 1 h. After the reaction was completed, the reaction mixture was quenched with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (200.0 mg, 50%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=529.2.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (189.0 mg, 0.36 mmol) in 1,4-dioxane (8.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (184.1 mg, 1.79 mmol), BrettPhos (38.3 mg, 0.07 mmol), Cs2CO3 (349.1 mg, 1.07 mmol) and BrettPhos Pd G3 (32.4 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was irradiated with microwave radiation at 120° C. for 1.5 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)-2-fluoro-N-(1-[[2-(trimethylsilyl)ethoxy]methyl]-3-(1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazol-5-yl)pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (166.0 mg, 78%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=596.3.
To a solution of (1S,2S)-2-fluoro-N-(1-[[2-(trimethylsilyl)ethoxy]methyl]-3-(1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazol-5-yl)pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (166.0 mg, 0.28 mmol) in DMF (8.0 mL) was added ethane-1,2-diamine (83.7 mg, 1.39 mmol) and TBAF (0.8 mL, 0.28 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 19% B to 39% B in 6 min; 254 nm) to (1S,2S)—N-[3-(1H-1,3-benzodiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (7.7 mg, 8%) as a white solid. LCMS (ESI, m/z): [M+H]+=336.1. 1H NMR (300 MHz, DMSO-d6): δ 12.40-12.35 (m, 1H), 11.63 (s, 1H), 10.78 (s, 1H), 8.26-8.19 (m, 2H), 7.91-7.89 (m, 2H), 7.77-7.70 (m, 2H), 7.56-7.54 (m, 1H), 5.01-4.78 (m, 1H), 2.59-2.50 (m, 1H), 1.54-1.46 (m, 1H), 1.28-1.22 (m, 1H).
To a solution of 5-bromo-2-methyl-1H-benzo[d]imidazole (500.0 mg, 2.37 mmol) in THF (20.0 mL) was added NaH (284.3 mg, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. Then SEM-Cl (592.4 mg, 3.55 mmol) was added dropwise to the mixture at 0° C. under N2. The resulting mixture was stirred at 0° C. for another 1 h. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (63/37, v/v) to afford 5-bromo-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole (710.0 mg, 87%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=341.1.
To a solution of 5-bromo-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole (660.0 mg, 1.93 mmol) in 1,4-dioxane (15.0 mL) was added 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (1473.1 mg, 5.80 mmol), KOAc (569.3 mg, 5.80 mmol) and Pd(dppf)Cl2 (141.5 mg, 0.19 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (45/55, v/v) to afford 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole (620.0 mg, 80%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=389.2.
To a solution of 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole (570.0 mg, 1.47 mmol) in 1,4-dioxane/H2O (20.0/4.0 mL) was added 3-bromo-6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (530.9 mg, 1.47 mmol), K2CO3 (608.5 mg, 4.40 mmol) and Pd(dppf)Cl2 (107.4 mg, 0.15 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (43/57, v/v) to afford 5-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole (790.0 mg, 99%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=543.2.
To a solution of 5-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazole (750.0 mg, 1.38 mmol) in 1,4-dioxane (15.0 mL) was added cyclopropanecarboxamide (587.5 mg, 6.90 mmol), Cs2CO3 (1349.4 mg, 4.14 mmol), BrettPhos (148.2 mg, 0.28 mmol) and BrettPhos Pd G3 (125.2 mg, 0.14 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (35/66, v/v) to afford N-(3-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-5-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (600.0 mg, 73%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=592.3.
To a solution of N-(3-(2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-5-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (550.0 mg, 0.93 mmol) in CH2Cl2 (10.0 mL) was added TFA (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (10.0 mL and NH3·H2O (10.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 16% B to 40% B in 8 min; 254 nm) to afford N-(3-(2-methyl-1H-benzo[d]imidazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (26.7 mg, 8%) as a white solid. LCMS (ESI, m/z): [M+H]+=332.2. 1H NMR (300 MHz, CD3OD): δ 8.25 (d, J=8.4 Hz, 1H), 7.85 (d, J=8.4 Hz, 1H), 7.74 (s, 1H), 7.57-7.51 (m, 3H), 2.60 (s, 3H), 1.94-1.89 (m, 1H), 1.05-0.96 (m, 2H), 0.94-0.91 (m, 2H).
To a mixture of 3-bromo-6-chloro-2-methoxypyridine (1.0 g, 4.49 mmol) and methylamine (1.4 g, 45.07 mmol) in NMP (15.0 mL) was added Cs2CO3 (4.3 g, 13.47 mmol) at room temperature. The resulting mixture was stirred at 120° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 5-bromo-6-methoxy-N-methylpyridin-2-amine (950.0 mg, 97%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=217.0.
To a mixture of 5-bromo-6-methoxy-N-methylpyridin-2-amine (900.0 mg, 4.14 mmol) and bis(pinacolato)diboron (3.1 g, 12.43 mmol) in dioxane (20.0 mL) were added Pd(dppf)Cl2 (303.3 mg, 0.41 mmol) and KOAc (1.2 g, 12.43 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 6-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (180.0 mg, 16%) as a white solid. LCMS (ESI, m/z): [M+H]+=265.2.
To a mixture of 6-methoxy-N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (160.0 mg, 0.62 mmol) and 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (150.0 mg, 0.41 mmol) in dioxane (4.0 mL) was added Pd(dppf)Cl2 (30.0 mg, 0.041 mmol) and K2CO3 (170.0 mg, 2.08 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-N-methylpyridin-2-amine (70.0 mg, 28%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=419.2.
To a mixture of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-N-methylpyridin-2-amine (40.0 mg, 0.09 mmol) and cyclopropanecarboxamide (32.5 mg, 0.38 mmol) in dioxane (3.0 mL) was added BrettPhos (10.2 mg, 0.02 mmol) BrettPhos Pd G3 (8.6 mg, 0.01 mmol) and Cs2CO3 (93.3 mg, 0.28 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford N-[3-[2-methoxy-6-(methylamino)pyridin-3-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (40.0 mg, 89%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=468.2.
To a solution of N-[3-[2-methoxy-6-(methylamino)pyridin-3-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (100.0 mg, 0.21 mmol) in DCM (2.0 mL) was added TFA (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added NH3·H2O (2.0 mL) and ACN (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 55% B in 10 min; 254 nm) to afford N-[3-[2-methoxy-6-(methylamino)pyridin-3-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (28.4 mg, 39%) as a white solid. LCMS (ESI, m/z): [M+H]+=338.2. 1H NMR (300 MHz, DMSO-d6): δ 11.36 (s, 1H), 10.53 (s, 1H), 7.96 (d, J=8.4 Hz, 1H), 7.85 (d, J=8.7 Hz, 1H), 7.61 (d, J=8.1 Hz, 1H), 7.42 (d, J=2.4 Hz, 1H), 6.39-6.34 (m, 1H), 6.10 (d, J=8.1 Hz, 1H), 3.87 (s, 3H), 2.82 (d, J=4.5 Hz, 3H), 2.06-2.01 (m, 1H), 0.83-0.75 (m, 4H).
To a solution of tert-butyl N-(5-bromo-4-methoxypyridin-2-yl)-N-(tert-butoxycarbonyl)carbamate (402.5 mg, 1.00 mmol) in dioxane/H2O (15.0/3.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (340.0 mg, 0.83 mmol), K2CO3 (344.8 mg, 2.50 mmol), Pd(dppf)Cl2 (67.9 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford tert-butyl N-(tert-butoxycarbonyl)-N-[5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxypyridin-2-yl]carbamate (330.0 mg, 66%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=605.2.
To a solution of tert-butyl N-(tert-butoxycarbonyl)-N-[5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxypyridin-2-yl]carbamate (282.0 mg, 0.47 mmol) in 1,4-dioxane (15.0 mL) was added cyclopropanecarboxamide (198.3 mg, 2.33 mmol), BrettPhos (50.0 mg, 0.03 mmol), Cs2CO3 (455.46 mg, 1.40 mmol) and BrettPhos Pd G3 (42.3 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was irradiated with microwave radiation at 120° C. for 1.5 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to tert-butyl N-(tert-butoxycarbonyl)-N-[5-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxypyridin-2-yl]carbamate (80.0 mg, 26%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=654.3.
To a solution of tert-butyl N-(tert-butoxycarbonyl)-N-[5-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxypyridin-2-yl]carbamate (80.0 mg, 0.12 mmol) in CH2Cl2 (4.0 mL) was added TFA (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 14% B to 36% B in 7 min; 254 nm) to afford N-[3-(6-amino-4-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (10.8 mg, 27%) as a white solid. LCMS (ESI, m/z): [M+H]+=324.2. 1H NMR (300 MHz, DMSO-d6): δ 11.33 (s, 1H), 10.47 (s, 1H), 7.84-7.78 (m, 3H), 7.29 (d, J=2.1 Hz, 1H), 6.09 (s, 1H), 5.78 (s, 2H), 3.70 (s, 3H), 1.96-1.92 (m, 1H), 0.75-0.68 (m, 4H).
To a solution of 5-bromo-6-methylpyridin-2-amine (5.0 g, 26.73 mmol) in CH2Cl2 (100.0 mL) was added Boc2O (5.8 g, 26.73 mmol) and DMAP (3.3 g, 26.73 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford tert-butyl (5-bromo-6-methylpyridin-2-yl)carbamate (4.0 g, 52%) as a white solid. LCMS (ESI, m/z): [M+H]+=287.0.
To a solution of tert-butyl (5-bromo-6-methylpyridin-2-yl)carbamate (2.0 g, 6.97 mmol) in dioxane (40.0 mL) was added 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (5.3 g, 20.89 mmol), KOAc (2.1 g, 20.89 mmol) and Pd(dppf)Cl2 (0.5 g, 0.70 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford tert-butyl (6-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)carbamate (2.0 g, 85%) as a white solid. LCMS (ESI, m/z): [M+H]+=335.2.
To a solution of tert-butyl (6-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)carbamate (500.0 mg, 1.50 mmol) in dioxane/H2O (10.0/1.0 mL) was added 3-bromo-6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (541.2 mg, 1.50 mmol), K2CO3 (620.3 mg, 4.49 mmol) and Pd(dppf)Cl2 (109.5 mg, 0.15 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford tert-butyl (5-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-6-methylpyridin-2-yl)carbamate (170.0 mg, 23%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=489.2.
To a solution of tert-butyl (5-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-6-methylpyridin-2-yl)carbamate (130.0 mg, 0.27 mmol) in dioxane (10.0 mL) was added cyclopropanecarboxamide (135.7 mg, 1.60 mmol), Cs2CO3 (259.8 mg, 0.80 mmol), Brettphos (28.5 mg, 0.05 mmol) and BrettPhos Pd G3 (24.1 mg, 0.03 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford tert-butyl (5-(6-(cyclopropanecarboxamido)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-6-methylpyridin-2-yl)carbamate (130.0 mg, 90%) as a white solid. LCMS (ESI, m/z): [M+H]+=538.3.
To a solution of tert-butyl (5-(6-(cyclopropanecarboxamido)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-6-methylpyridin-2-yl)carbamate (250.0 mg, 0.47 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 25 mL/min; Gradient: 6% to 17% in 7 min; 254 nm) to afford N-(3-(6-amino-2-methylpyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (27.8 mg, 19%) as a white solid. LCMS (ESI, m/z): [M+H]+=308.2. 1H NMR (400 MHz, DMSO-d6): δ 11.46 (s, 1H), 10.57 (s, 1H), 7.86 (d, J=8.8 Hz, 1H), 7.73 (d, J=8.4 Hz, 1H), 7.36-7.30 (m, 2H), 6.38 (d, J=8.4 Hz, 1H), 5.86 (s, 2H), 2.27 (s, 3H), 2.07-2.01 (m, 1H), 0.84-0.81 (m, 4H).
To a solution of 5-bromo-6-methylpyridin-2-amine (10.0 g, 53.46 mmol) in THF (100.0 mL) was added NaH (1.9 g, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h under N2. Then CH3I (7.6 g, 53.46 mmol) was added dropwise to the mixture at 0° C. under N2. The resulting mixture was stirred at 0° C. for another 1 h. After the reaction was completed, the reaction was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 5-bromo-N,6-dimethylpyridin-2-amine (3.3 g, 30%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=201.0.
To a solution of 5-bromo-N,6-dimethylpyridin-2-amine (2.0 g, 9.95 mmol) in dioxane (40.0 mL) was added 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (7.6 g, 29.84 mmol), KOAc (8.9 g, 29.84 mmol) and Pd(dppf)Cl2 (0.7 g, 1.00 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/9, v/v) to afford N,6-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (750.0 mg, 30%) as a brown oil. LCMS (ESI, m/z): [M+H]+=249.2.
To a solution of N,6-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (500.0 mg, 2.02 mmol) in dioxane/H2O (10.0/1.0 mL) was added 3-bromo-6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (728.9 mg, 2.02 mmol), K2CO3 (835.5 mg, 6.05 mmol) and Pd(dppf)Cl2 (147.4 mg, 0.20 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 5-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-N,6-dimethylpyridin-2-amine (250.0 mg, 30%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=403.2.
To a solution of 5-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-N,6-dimethylpyridin-2-amine (200.0 mg, 0.50 mmol) in dioxane (10.0 mL) was added cyclopropanecarboxamide (253.4 mg, 2.98 mmol), Cs2CO3 (485.1 mg, 1.49 mmol), Brettphos (53.3 mg, 0.10 mmol) and BrettPhos Pd G3 (45.0 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford N-(3-(2-methyl-6-(methylamino)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (150.0 mg, 66%) as a white solid. LCMS (ESI, m/z): [M+H]+=452.2.
To a solution of N-(3-(2-methyl-6-(methylamino)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (130.0 mg, 0.29 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 24% to 44% in 7 min; 254 nm) to afford N-(3-(2-methyl-6-(methylamino)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (44.1 mg, 47%) as a white solid. LCMS (ESI, m/z): [M+H]+=322.2. 1H NMR (300 MHz, DMSO-d6): δ 11.46 (d, J=1.5 Hz, 1H), 10.57 (s, 1H), 7.87 (d, J=8.7 Hz, 1H), 7.73 (d, J=8.4 Hz, 1H), 7.36 (d, J=8.4 Hz, 1H), 7.29 (d, J=2.4 Hz, 1H), 6.36-6.31 (m, 2H), 2.79 (d, J=4.8 Hz, 3H), 2.30 (s, 3H), 2.08-2.00 (m, 1H), 0.85-0.75 (m, 4H).
To a solution of tert-butyl N-(5-bromo-4-methylpyridin-2-yl)carbamate (500.0 mg, 1.71 mmo) in dioxane (10.0 mL) was added 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (1326.4 mg, 5.24 mmol), Pd(dppf)Cl2 (254.8 mg, 0.38 mmol) and KOAc (512.6 mg, 5.24 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/2, v/v) to afford 6-[(tert-butoxycarbonyl)amino]-4-methylpyridin-3-ylboronic acid (400.0 mg, 69%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=253.1.
To a solution of 6-[(tert-butoxycarbonyl)amino]-4-methylpyridin-3-ylboronic acid (300.0 mg, 1.19 mmol) in dioxane/H2O (10.0 mL/1.0 mL) was added 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (344.4 mg, 0.95 mmol), Pd(dppf)Cl2 (87.0 mg, 0.19 mmol) and K2CO3 (493.4 mg, 3.57 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford tert-butyl N-[5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methylpyridin-2-yl]carbamate (130.0 mg, 22%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=489.2.
To a solution of tert-butyl N-[5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methylpyridin-2-yl]carbamate (110.0 mg, 0.25 mmol) in dioxane (10.0 mL) was added cyclopropanecarboxamide (57.4 mg, 0.65 mmol), BrettPhos Pd G3 (20.9 mg, 0.02 mmol), BrettPhos (24.4 mg, 0.05 mmol) and Cs2CO3 (219.8 mg, 0.67 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford tert-butyl N-[5-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methylpyridin-2-yl]carbamate (78.0 mg, 64%) as a white oil. LCMS (ESI, m/z): [M+H]+=538.3.
To a solution of tert-butyl N-[5-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methylpyridin-2-yl]carbamate (78.0 mg, 0.14 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue washed with CH2Cl2 and then filtered. The solid was collected and dried to afford N-[3-(6-amino-4-methylpyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (10.0 mg, 22%) as a white solid. LCMS (ESI, m/z): [M+H]+=308.1. 1H NMR (300 MHz, DMSO-d6): δ 11.82 (s, 1H), 10.72 (s, 1H), 8.02-7.90 (m, 3H), 7.64-7.53 (m, 3H), 6.93 (s, 1H), 2.37 (s, 3H), 2.13-2.07 (m, 1H), 0.93-0.85 (m, 4H).
To a solution of 5-bromo-4-methylpyridin-2-amine (5.0 g, 26.72 mmol) in THF (100.0 mL) was added di-tert-butyl dicarbonate (5.8 g, 26.72 mmol) and DMAP (3.7 g, 26.72 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (6/1, v/v) to afford tert-butyl N-(5-bromo-4-methylpyridin-2-yl)carbamate (2.7 g, 35%) as a white solid. LCMS (ESI, m/z): [M+H]+=287.0
To a solution of tert-butyl N-(5-bromo-4-methylpyridin-2-yl)carbamate (700.0 mg, 2.48 mmol) in THF (10.0 mL) was added NaH (117.0 mg, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. Then CH3I (692.0 mg, 4.88 mmol) was added to the mixture. The resulting mixture was stirred at 0° C. for another 1 h. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford tert-butyl N-(5-bromo-4-methylpyridin-2-yl)-N-methylcarbamate (600.0 mg, 95%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=301.0.
To a solution of tert-butyl N-(5-bromo-4-methylpyridin-2-yl)-N-methylcarbamate (600.0 mg, 1.92 mmol) in dioxane (10.0 mL, 510.75 mmol) was added 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (1.5 g, 5.98 mmol), Pd(dppf)Cl2 (145.7 mg, 0.19 mmol) and KOAc (586.5 mg, 5.97 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (7/1, v/v) to afford tert-butyl methyl(4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)carbamate (500.0 mg, 72%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=349.2.
To a solution of tert-butyl methyl(4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)carbamate (400.0 mg, 1.50 mmol) in dioxane/H2O (10.0/1.0 mL) was added 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (1.1 g, 3.06 mmol), Pd(dppf)Cl2 (109.9 mg, 0.15 mmol) and K2CO3 (623.5 mg, 4.50 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford tert-butyl N-[5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methylpyridin-2-yl]-N-methylcarbamate (400.0 mg, 53%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=503.2.
To a solution of tert-butyl N-[5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methylpyridin-2-yl]-N-methylcarbamate (380.0 mg, 0.75 mmol) in dioxane (10.0 mL) was added cyclopropanecarboxamide (192.8 mg, 2.26 mmol), BrettPhos Pd G3 (68.7 mg, 0.07 mmol), BrettPhos (81.0 mg, 0.15 mmol) and Cs2CO3 (738.6 mg, 2.26 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford tert-butyl N-[5-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methylpyridin-2-yl]-N-methylcarbamate (300.0 mg, 72%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=552.3.
To a solution of tert-butyl N-[5-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methylpyridin-2-yl]-N-methylcarbamate (150.0 mg, 0.27 mmol) in DCM (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 16% B to 46% B in 10 min; 254 nm) to afford N-[3-[4-methyl-6-(methylamino)pyridin-3-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (17.4 mg, 20%) as a white solid. LCMS (ESI, m/z): [M+H]+=322.2. 1H NMR (300 MHz, DMSO-d6): δ 11.47 (s, 1H), 10.56 (s, 1H), 7.92 (s, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.73 (d, J=8.7 Hz, 1H), 7.30 (d, J=2.1 Hz, 1H), 6.39 (s, 1H), 6.32-6.28 (m, 1H), 2.79 (d, J=4.8 Hz, 3H), 2.14 (s, 3H), 2.05-2.01 (m, 1H), 0.85-0.77 (m, 4H).
To a solution of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (500.0 mg, 2.27 mmol) in dioxane/H2O (5.0/0.5 mL) was added 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (986.2 mg, 2.72 mmol), Pd(dppf)Cl2 (332.4 mg, 0.45 mmol) and K2CO3 (941.9 mg, 6.81 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (62/38, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-amine (150.0 mg, 44%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=375.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-amine (330.0 mg, 0.88 mmol) in DCM (5.0 mL) was added di-tert-butyl dicarbonate (768.3 mg, 3.52 mmol), TEA (267.1 mg, 2.64 mmol) and DMAP (21.5 mg, 0.17 mmol) at room temperature. The resulting mixture was stirred at room temperature for 5 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (50/50, v/v) to afford tert-butyl N-[3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl]carbamate (460.0 mg, 72%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=575.2.
To a solution of tert-butyl N-(tert-butoxycarbonyl)-N-[3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl]carbamate (440.0 mg, 0.76 mmol) in dioxane (5.0 mL) was added cyclopropanecarboxamide (325.5 mg, 3.82 mmol), BrettPhos (164 mg, 0.30 mmol), BrettPhos Pd G3 (138.6 mg, 0.15 mmol) and Cs2CO3 (747.7 mg, 2.29 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 5 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (76/24, v/v) to afford tert-butyl N-(tert-butoxycarbonyl)-N-[3-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl]carbamate (360.0 mg, 75%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=624.3.
To a solution of tert-butyl N-(tert-butoxycarbonyl)-N-[3-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl]carbamate (310.0 mg, 0.49 mmol) in DCM (2.0 mL) was added TFA (2.0 mL). The mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in ACN (2.0 mL) and NH3·H2O (2.0 mL). The mixture was stirred at room temperature for another 5 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 45% B to 75% B in 7 min; 254 nm) to afford N-[3-(2-aminopyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (48.4 mg, 33%) as a white solid. LCMS (ESI, m/z): [M+H]+=294.1. 1H NMR (300 MHz, DMSO-d6): δ 10.62 (s, 1H), 7.94-7.83 (m, 3H), 7.53-7.47 (m, 2H), 6.68-6.64 (m, 1H), 6.04 (s, 1H), 5.56 (s, 2H), 2.08-2.00 (m, 1H), 0.86-0.81 (m, 4H).
To a solution of 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (220.0 mg, 0.53 mmol) in 1,4-dioxane/H2O (4.0/0.4 mL) was added tert-butyl N-(3-bromopyridin-2-yl)-N-methylcarbamate (154.5 mg, 0.53 mmol), Pd(dppf)Cl2 (78.7 mg, 0.10 mmol) and K2CO3 (223.1 mg, 1.61 mmol) at room temperature under N2, The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (70/30, v/v) to afford tert-butyl N-[3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl]-N-methylcarbamate (140.0 mg, 53%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=489.2
To a solution of tert-butyl N-[3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl]-N-methylcarbamate (140.0 mg, 0.28 mmol) in dioxane (4.0 mL) was added cyclopropanecarboxamide (121.8 mg, 1.43 mmol), BrettPhos (61.5 mg, 0.11 mmol), BrettPhos Pd G3 (51.9 mg, 0.11 mmol) and Cs2CO3 (279.8 mg, 0.56 mmol) at room temperature under N2, The resulting mixture was stirred at 100° C. for 5 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (70/30, v/v) to afford tert-butyl N-[3-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl]-N-methylcarbamate (120.0 mg, 77%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=538.3.
To a solution of tert-butyl N-[3-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl]-N-methylcarbamate (200.0 mg, 0.37 mmol) in DCM (2.0 mL) was added TFA (0.5 mL). The mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in NH3·H2O (0.5 mL) and CH3CN (2.0 mL). The mixture was stirred at room temperature for another 6 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (75/25, v/v) and then purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 27% B to 57% B in 7 min; 254 nm) to afford N-[3-[2-(methylamino)pyridin-3-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (10.7 mg, 98%) as a white solid. LCMS (ESI, m/z): [M+H]+=308.1. 1H NMR (300 MHz, DMSO-d6): δ 11.65 (s, 1H), 10.59 (s, 1H), 8.02-8.00 (m, 1H), 7.90-7.84 (m, 1H), 7.77 (d, J=8.7 Hz, 1H), 7.50 (s, 1H), 7.41-7.38 (m, 1H), 6.62-6.58 (m, 1H), 5.76-5.73 (m, 1H), 2.79 (d, J=4.5 Hz, 3H), 2.05-1.99 (m, 1H), 0.92-0.75 (m, 4H).
To a solution of 6-bromopyridin-2-amine (5.0 g, 28.90 mmol) in CH2Cl2 (100.0 mL) was added Boc2O (6.3 g, 28.90 mmol) and DMAP (3.5 g, 28.90 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (88/12, v/v) to afford tert-butyl (6-bromopyridin-2-yl)carbamate (4.3 g, 54%) as a white solid. LCMS (ESI, m/z): [M+H]+=273.0.
To a solution of 3-bromo-6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (1.0 g, 1.55 mmol) in 1,4-dioxane (15.0 mL) was added 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (1.2 g, 4.65 mmol), KOAc (460.0 mg, 4.65 mmol) and Pd(dppf)Cl2 (113.3 mg, 0.16 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (92/8, v/v) to afford 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (330.0 mg, 52%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=409.2.
To a solution of 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (300.0 mg, 0.73 mmol) in 1,4-dioxane/H2O (10.0/2.0 mL) was added tert-butyl (6-bromopyridin-2-yl)carbamate (200.4 mg, 0.73 mmol), K2CO3 (304.3 mg, 2.20 mmol) and Pd(dppf)Cl2 (53.7 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (92/8, v/v) to afford tert-butyl (6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl)carbamate (150.0 mg, 43%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=475.2.
To a solution of tert-butyl (6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl)carbamate (130.0 mg, 0.27 mmol) in 1,4-dioxane (10.0 mL) was added cyclopropanecarboxamide (116.5 mg, 1.37 mmol), Cs2CO3 (267.5 mg, 0.82 mmol), BrettPhos (29.4 mg, 0.06 mmol) and BrettPhos Pd G3 (24.8 mg, 0.03 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (75/25, v/v) to afford tert-butyl (6-(6-(cyclopropanecarboxamido)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl)carbamate (140.0 mg, 97%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=524.3.
To a solution of tert-butyl (6-(6-(cyclopropanecarboxamido)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl)carbamate (120.0 mg, 0.23 mmol) in CH2Cl2 (6.0 mL) was added TFA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in CH3CN (6.0 mL) was added NH3·H2O (6.0 mL). The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The solid was washed with CH2Cl2 and H2O to afford N-(3-(6-aminopyridin-2-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (31.1 mg, 46%) as a white solid. LCMS (ESI, m/z): [M+H]+=294.1. 1H NMR (300 MHz, DMSO-d6): δ 11.60 (s, 1H), 10.58 (s, 1H), 8.77 (d, J=8.7 Hz, 1H), 7.95-7.91 (m, 2H), 7.39-7.34 (m, 1H), 6.99 (d, J=7.5 Hz, 1H), 6.25 (d, J=8.1 Hz, 1H), 5.86 (s, 2H), 2.08-2.00 (m, 1H), 0.83-0.79 (m, 4H).
To a solution of tert-butyl (6-bromopyridin-2-yl)carbamate (1.0 g, 3.67 mmol) in DMF (30.0 mL) was added NaH (439.3 mg, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. Then CH3I (2.6 g, 18.31 mmol) was added dropwise to the mixture at 0° C. The resulting mixture was stirred at 0° C. for another 1 h. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (91/9, v/v) to afford tert-butyl (6-bromopyridin-2-yl)(methyl)carbamate (1.0 g, 95%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=287.0.
To a solution of 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (440.0 mg, 1.07 mmol) in 1,4-dioxane/H2O (10.0/2.0 mL) was added tert-butyl (6-bromopyridin-2-yl)(methyl)carbamate (309.0 mg, 1.07 mmol), K2CO3 (448.1 mg, 3.21 mmol) and Pd(dppf)Cl2 (78.6 mg, 0.11 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (92/8, v/v) to afford tert-butyl (6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl)(methyl)carbamate (270.0 mg, 53%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=489.2.
To a solution of tert-butyl (6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl)(methyl)carbamate (250.0 mg, 0.51 mmol) in 1,4-dioxane (10.0 mL) was added cyclopropanecarboxamide (217.5 mg, 2.56 mmol), Cs2CO3 (499.6 mg, 1.53 mmol), BrettPhos (54.9 mg, 0.10 mmol) and BrettPhos Pd G3 (46.3 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (75/25, v/v) to afford tert-butyl (6-(6-(cyclopropanecarboxamido)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl)(methyl)carbamate (240.0 mg, 87%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=538.3.
To a solution of tert-butyl (6-(6-(cyclopropanecarboxamido)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl)(methyl)carbamate (200.0 mg, 0.37 mmol) in CH2Cl2 (6.0 mL) was added TFA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in CH3CN (6.0 mL) was added NH3·H2O (6.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The solid was washed with CH2Cl2 and H2O. The solid was collected and dried to afford N-(3-(6-(methylamino)pyridin-2-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (47.0 mg, 41%) as a white solid. LCMS (ESI, m/z): [M+H]+=308.2. 1H NMR (300 MHz, DMSO-d6) δ 11.61 (s, 1H), 10.58 (s, 1H), 8.77 (d, J=8.7 Hz, 1H), 7.94-7.92 (m, 2H), 7.40-7.35 (m, 1H), 6.99 (d, J=7.2 Hz, 1H), 6.40-6.39 (m, 1H), 6.24 (d, J=8.1 Hz, 1H), 2.89 (d, J=4.2 Hz, 3H), 2.08-2.00 (m, 1H), 0.83-0.79 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (1.0 g, 2.76 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (0.7 g, 3.31 mmol), K2CO3 (1.2 g, 8.29 mmol) and Pd(dppf)Cl2 (0.5 g, 0.55 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (93/7, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-amine (300.0 mg, 29%) as a white solid. LCMS (ESI, m/z): [M+H]+=375.1.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-amine (300.0 mg, 0.85 mmol) in dioxane (3.0 mL) was added cyclopropanecarboxamide (363.2 mg, 4.27 mmol), BrettPhos (183.3 mg, 0.34 mmol), Cs2CO3 (834.2 mg, 2.56 mmol) and BrettPhos Pd G3 (154.7 mg, 0.17 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (40/60, v/v) to afford N-[3-(6-aminopyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (38.0 mg, 10%) as a white solid. LCMS (ESI, m/z): [M+H]+=424.2.
To a solution of N-[3-(6-aminopyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (38.0 mg, 0.09 mmol) in DMF (2.0 mL) was added ethylenediamine (2.0 mL) and TBAF (2.0 mL) at room temperature. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 27% B to 49% B in 9 min; 254 nm to afford N-[3-(6-aminopyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (2.3 mg, 9%) as a white solid. LCMS (ESI, m/z): [M+H]+=294.2. 1H NMR (400 MHz, DMSO-d6): δ 11.64 (s, 1H), 10.61 (s, 1H), 8.24 (s, 1H), 8.18-8.13 (m, 1H), 8.03 (d, J=8.4 Hz, 1H), 7.93 (d, J=8.4 Hz, 1H), 7.67 (s, 1H), 7.09-6.68 (m, 3H), 2.07-1.99 (m, 1H), 0.82-0.78 (m, 4H).
To a solution of cis-2-(hydroxymethyl)-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (290.0 mg, 0.62 mmol) in DCM (10.0 mL) was added Dess-Martin (394.5 mg, 0.93 mmol). The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum to afford cis-2-formyl-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (280.0 mg, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=466.2.
To a solution of cis-2-formyl-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (300.0 mg, crude) in CH2Cl2 (10.0 mL) was added 1-methylpiperazine (129.1 mg, 1.29 mmol) and NaBH3CN (81.0 mg, 1.29 mmol). The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by reverse phase flash chromatography with CH3OH/H2O (60/40, v/v) to afford cis-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (280.0 mg, 79%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=550.3.
To a solution of cis-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (280.0 mg, 0.51 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL). The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. The residue was dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 47% B in 10 min; 254 nm) to afford cis-N-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (40.7 mg, 19%) as a white solid. LCMS (ESI, m/z): [M+H]+=420.2. 1H NMR (300 MHz, DMSO-d6): δ 11.53 (s, 1H), 10.51 (s, 1H), 7.97 (d, J=8.7 Hz, 1H), 7.88 (d, J=8.7 Hz, 1H), 7.56-7.52 (m, 2H), 7.30-7.25 (m, 1H), 7.11 (d, J=7.5 Hz, 1H), 7.05-7.00 (m, 1H), 3.82 (s, 3H), 2.60-2.52 (m, 1H), 2.49-2.40 (m, 5H), 2.28-2.20 (m, 4H), 2.15-2.09 (m, 4H), 1.35-1.30 (m, 1H), 1.02-0.98 (m, 1H), 0.90-0.87 (m, 1H).
To a solution of 6-chloro-3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (1.9 g, 4.89 mmol) in 1,4-dioxane (20.0 mL) was added diphenylmethanimine (2.7 g, 14.65 mmol), Cs2CO3 (4.8 g, 14.65 mmol), BrettPhos (0.5 g, 0.98 mmol) and BrettPhos Pd G3 (0.4 g, 0.49 mmol). The resulting mixture was stirred at 100° C. for 2 h under N2. After the reaction was completed, the mixture was concentrated under vacuum. The residue was purified by flash column chromatography with ether/ethyl acetate (20/80, v/v) to afford N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1,1-diphenylmethanimine (900.0 mg, 34%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=534.2.
The solution of N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1,1-diphenylmethanimine (900.0 mg, 1.69 mmol) in HCl/1,4-dioxane (3.0 mL, 4 mol/L) was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was washed with Et2O to afford 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine hydrochloride (400.0 mg, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=370.2.
To a solution of 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine hydrochloride (282.6 mg, 3.57 mmol) in CH2Cl2 (3.0 mL) was added pyridine (282.6 mg, 3.57 mmol) and phenyl chloroformate (167.8 mg, 1.07 mmol) at 0° C. The mixture was stirred at room temperature for 16 h. The mixture was concentrated under vacuum. Then pyridine (2.5 mL) and 2-(4-methylpiperazin-1-yl)ethanamine (255.8 mg, 1.79 mmol) were added to the residue. The resulting mixture was stirred at 60° C. for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (20/80, v/v) to afford 3-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (200.0 mg, 42%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=539.3.
To a solution of 1-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(2-(4-methylpiperazin-1-yl)ethyl)urea (180.0 mg, 0.33 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL). The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. Then ACN (2.0 mL) and NH3·H2O (2.0 mL) were added to the residue. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18 Column, 20×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 27% B to 57% B in 10 min; 254 nm) to afford 3-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (18.8 mg, 14%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=409.2. 1H NMR (300 MHz, CDCl3): δ 10.07 (s, 1H), 9.73 (s, 1H), 7.94 (d, J=8.4 Hz, 1H), 7.55-7.52 (m, 1H), 7.45-7.42 (m, 2H), 7.35-7.28 (m, 1H), 7.10-7.04 (m, 2H), 6.48 (d, J=8.4 Hz, 1H), 3.90 (s, 3H), 3.67-3.61 (m, 2H), 2.88-2.83 (m, 6H), 2.62-2.59 (m, 4H), 2.40 (s, 3H).
To a solution of 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (250.0 mg, 0.68 mmol) in CH2Cl2 (5.0 mL) was added Pyridine (107.0 mg, 1.35 mmol) and phenyl chloroformate (127.1 mg, 0.81 mmol) at 0° C. The mixture was stirred at room temperature for 16 h. The mixture was concentrated under vacuum. Then pyridine (2.5 mL) and 2-morpholinoethanamine (264.2 mg, 2.03 mmol) were added to the residue. The resulting mixture was stirred at 60° C. for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (20/80, v/v) to afford 1-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(2-morpholinoethyl)urea (200.0 mg, 42%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=526.3.
To a solution of 3-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(morpholin-4-yl)ethyl]urea (120.0 mg, 0.23 mmol) in CH2Cl2 (2.0 mL) was added TFA (1.0 mL). The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. Then ACN (2.0 mL) and NH3·H2O (2.0 mL) were added to the mixture. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column, 20×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 42% B in 10 min; 254 nm) to afford 3-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(morpholin-4-yl)ethyl]urea (31.5 mg, 35%) as a white solid. LCMS (ESI, m/z): [M+H]+=396.2. 1H NMR (300 MHz, DMSO-d6): δ 11.40 (s, 1H), 9.19 (s, 1H), 8.37 (s, 1H), 7.91 (d, J=8.7 Hz, 1H), 7.53-7.46 (m, 2H), 7.29-7.23 (m, 1H), 7.12-6.99 (m, 3H), 3.81 (s, 3H), 3.61-3.58 (m, 4H), 3.38-3.34 (m, 2H), 2.47-2.42 (m, 6H).
A mixture of 6-chloro-1H-pyrazolo[3,4-b]pyridine (1.0 g, 6.51 mmol) and NIS (2.3 g, 0.01 mmol) in DMF (20.0 mL) was stirred at 110° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum to afford 6-chloro-3-iodo-1H-pyrazolo[3,4-b]pyridine (1.7 g, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=279.9.
To a solution of 6-chloro-3-iodo-1H-pyrazolo[3,4-b]pyridine (2.7 g, 9.63 mmol) in THF (20.0 mL) was added NaH (0.7 g, 60%) at 0° C. under N2. The mixture was stirred at 0° C. for 1 h under N2. Then SEM-Cl (2.4 g, 14.44 mmol) was added to the mixture. The mixture was stirred at room temperature for another 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (14/86, v/v) to afford 6-chloro-3-iodo-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (750.0 mg, 46%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=410.0.
A mixture of 6-chloro-3-iodo-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (700.0 mg, 1.71 mmol), 2-methoxyphenylboronic acid (259.6 mg, 1.71 mmol), K2CO3 (708.4 mg, 5.13 mmol) and Pd(dppf)Cl2 (125.0 mg, 0.17 mmol) in dioxane (10.0 mL) and H2O (2.0 mL) was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (87/13, v/v) to afford 6-chloro-3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (444.0 mg, 67%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=390.1.
A mixture of 6-chloro-3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (440.0 mg, 1.13 mmol), cyclopropanecarboxamide (288.1 mg, 3.39 mmol), Cs2CO3 (1102.9 mg, 3.39 mmol), BrettPhos (121.1 mg, 0.23 mmol) and BrettPhos Pd G3 (102.3 mg, 0.11 mmol) in dioxane (20.0 mL) was stirred at 100° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (84/16, v/v) to afford N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropanecarboxamide (483.0 mg, 97%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=439.2.
A mixture of N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropanecarboxamide (550.0 mg, 1.25 mmol) and TFA (5.0 mL, 43.85 mmol) in CH2Cl2 (2.0 mL) was stirred at room temperature for 2 h. The mixture was evaporated in vacuo. The residue was dissolved in NH3·H2O (2.0 mL) and CH3CN (5.0 mL). The mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (94/6, v/v) and then purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 23% B to 53% B in 10 min; 254 nm) to afford N-[3-(2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-6-yl]cyclopropanecarboxamide (56.8 mg, 15%) as a white solid. LCMS (ESI, m/z): [M+H]+=309.0. 1H NMR (300 MHz, DMSO-d6): δ 13.42 (s, 1H), 10.89 (s, 1H), 8.08 (d, J=9.0 Hz, 1H), 7.98 (d, J=9.0 Hz, 1H), 7.63-7.60 (m, 1H), 7.46-7.40 (m, 1H), 7.18 (d, J=7.8 Hz, 1H), 7.09-7.04 (m, 1H), 3.83 (s, 3H), 2.08-2.04 (m, 1H), 0.85-0.82 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (600.0 mg, 1.66 mmol) in 1,4-dioxane/H2O (10.0/2.0 mL) was added 2,6-dimethoxyphenylboronic acid (362.2 mg, 1.99 mmol), K2CO3 (458.5 mg, 3.32 mmol) and Pd(dppf)Cl2 (121.4 mg, 0.17 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h under N2. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford 6-chloro-3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (93.0 mg, 13%) as a brown oil. LCMS (ESI, m/z): [M+H]+=419.1.
To a solution of 6-chloro-3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (93.0 mg, 0.22 mmol) in 1,4-dioxane (4.0 mL) was added cyclopropanecarboxamide (94.5 mg, 1.11 mmol), Cs2CO3 (217.0 mg, 0.67 mmol), BrettPhos (23.8 mg, 0.04 mmol) and BrettPhos Pd G3 (20.1 mg, 0.02 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford N-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (100.0 mg, 96%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=468.2.
To a solution of N-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (260.0 mg, 0.56 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Shield RP18 OBD Column, 19×250 mm, 10 um; Mobile Phase A: Water (10 mmol/L NH4HCO3+0.1% NH3·H2O), Mobile Phase B: MeOH—Preparative; Flow rate: 25 mL/min; Gradient: 50% B to 65% B in 9 min; 254 nm) to afford N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (11.9 mg, 6%) as a white solid. LCMS (ESI, m/z): [M+H]+=338.1. 1H NMR (300 MHz, DMSO-d6): δ 11.40 (s, 1H), 10.52 (s, 1H), 7.81 (d, J=8.7 Hz, 1H), 7.53 (d, J=8.4 Hz, 1H), 7.27-7.25 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 3.69 (s, 6H), 2.06-1.99 (m, 1H), 0.82-0.80 (m, 4H).
To a solution of 3-bromo-6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (1.0 g, 2.76 mmol) in 1,4-dioxane/H2O (20.0/4.0 mL) was added (2,6-difluorophenyl)boronic acid (436.5 mg, 2.76 mmol), K2CO3 (1.1 g, 8.29 mmol) and Pd(dppf)Cl2 (202.3 mg, 0.28 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (88/12, v/v) to afford 6-chloro-3-(2,6-difluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (160.0 mg, 12%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=395.1.
To a solution of 6-chloro-3-(2,6-difluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (250.0 mg, 0.63 mmol) in 1,4-dioxane (10.0 mL) was added cyclopropanecarboxamide (269.4 mg, 3.17 mmol), Cs2CO3 (618.8 mg, 1.90 mmol), BrettPhos (68.0 mg, 0.13 mmol) and BrettPhos Pd G3 (57.4 mg, 0.06 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (83/17, v/v) to afford N-(3-(2,6-difluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (50.0 mg, 18%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=444.2.
To a solution of N-(3-(2,6-difluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (50.0 mg, 0.11 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (2.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge BEH C18 OBD Prep Column, 5 μm, 19 mm 250 mm; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL/min; Gradient: 47% B to 51% B in 12 min; 254 nm) to afford N-(3-(2,6-difluorophenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (11.2 mg, 32%) as a white solid. LCMS (ESI, m/z): [M+H]+=314.1. 1H NMR (400 MHz, DMSO-d6): δ 11.86 (s, 1H), 10.61 (s, 1H), 7.90 (d, J=8.4 Hz, 1H), 7.76 (d, J=8.4 Hz, 1H), 7.59 (d, J=1.2 Hz, 1H), 7.46-7.34 (m, 1H), 7.22-7.18 (m, 2H), 2.10-1.98 (m, 1H), 0.82-0.76 (m, 4H).
To a mixture of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.82 mmol) and 2-ethoxypyridin-3-ylboronic acid (207.7 mg, 1.24 mmol) in dioxane/H2O (5.0/0.5 mL) was added Pd(dppf)Cl2 (60.6 mg, 0.08 mmol) and K2CO3 (343.8 mg, 2.48 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-ethoxypyridine (230.0 mg, 69%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=404.1.
To a mixture of 6-chloro-3-(2-ethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (260.0 mg, 0.62 mmol) and cyclopropanecarboxamide (211.2 mg, 2.48 mmol) in dioxane (4.0 mL) was added Brettphos Pd G3 (56.2 mg, 0.06 mmol), BrettPhos (66.6 mg, 0.12 mmol) and Cs2CO3 (606.5 mg, 1.82 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford N-(3-(2-ethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (260.0 mg, 89%) as a white solid. LCMS (ESI, m/z): [M+H]+=453.2
To a solution of N-[3-(2-ethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b] pyridin-6-yl]cyclopropanecarboxamide (270.0 mg, 0.59 mmol) in DCM (4.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the above mixture was added NH3·H2O (4.0 mL) and ACN (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 32% B to 62% B in 10 min; 254 nm) to afford N-[3-(2-ethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (46.7 mg, 24%) as a white solid. LCMS (ESI, m/z): [M+H]+=323.2. 1H NMR (300 MHz, DMSO-d6): δ 11.68 (s, 1H), 10.62 (s, 1H), 8.13 (d, J=9.0 Hz, 1H), 8.07-8.04 (m, 1H), 8.01-7.92 (m, 2H), 7.74 (d, J=2.7 Hz, 1H), 7.08-7.04 (m, 1H), 4.45-4.38 (m, 2H), 2.07-2.03 (m, 1H), 1.38-1.33 (m, 3H), 0.84-0.79 (m, 4H).
To a solution of 3-bromopyridin-2-ol (5.0 g, 28.73 mmol) in ACN (10.0 mL) was added 2,2-difluoro-2-(fluorosulfonyl)acetic acid (6.1 g, 34.48 mmol) and Na2SO4 (4.9 g, 34.48 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (94/6, v/v) to afford 3-bromo-2-(difluoromethoxy)pyridine (3.2 g, 50%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=223.9.
To a solution of 3-bromo-2-(difluoromethoxy)pyridine (200.0 mg, 0.89 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (574.5 mg, 1.34 mmol), K2CO3 (370.2 mg, 2.68 mmol) and Pd(dppf)Cl2 (145.8 mg, 0.18 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (83/17, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-(difluoromethoxy)pyridine (200.0 mg, 52%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=426.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-(difluoromethoxy)pyridine (200.0 mg, 0.47 mmol) in dioxane (3.0 mL) was added cyclopropanecarboxamide (119.9 mg, 1.41 mmol), BrettPhos (50.4 mg, 0.09 mmol), Cs2CO3 (459.0 mg, 1.41 mmol) and BrettPhos Pd G3 (42.6 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (88/12, v/v) to afford N-[3-[2-(difluoromethoxy)pyridin-3-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (200.0 mg, 89%) as a white solid. LCMS (ESI, m/z): [M+H]+=475.2.
To a solution of N-[3-[2-(difluoromethoxy)pyridin-3-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (190.0 mg, 0.40 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (2.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 34% B to 64% B in 10 min; 254 nm) to afford N-[3-[2-(difluoromethoxy)pyridin-3-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (45.2 mg, 28%) as a white solid. LCMS (ESI, m/z): [M+H]+=345.2. 1H NMR (400 MHz, DMSO-d6): δ 11.83 (s, 1H), 10.64 (s, 1H), 8.16 (d, J=6.0 Hz, 2H), 8.10 (d, J=8.8 Hz, 1H), 7.99-7.63 (m, 3H), 7.37-7.34 (m, 1H), 2.09-1.99 (m, 1H), 0.83-0.79 (m, 4H).
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (200.0 mg, 0.18 mmol) in 1,4-dioxane (8.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (92.2 mg, 0.89 mmol), BrettPhos (19.2 mg, 0.04 mmol), Cs2CO3 (174.8 mg, 0.54 mmol) and BrettPhos Pd G3 (16.2 mg, 0.02 mmol) at room temperature under N2. The resulting mixture was stirred with microwave at 100° C. for 2 h under N2. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/9, v/v) to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (50.0 mg, 45%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=626.3.
To a solution of (1S,2S)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (215.0 mg, 0.34 mmol) in DCM (6.0 mL) was added TFA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (2.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 53% B to 43% B in 7 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1H-1,3-benzodiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (6.9 mg, 6%) as a white solid. LCMS (ESI, m/z): [M+H]+=366.1. 1H NMR (300 MHz, DMSO-d6): δ 12.35-12.26 (br, 1H), 11.50 (s, 1H), 10.61 (s, 1H), 8.16 (s, 1H), 7.96-7.87 (m, 2H), 7.64 (s, 1H), 7.52 (d, J=2.1 Hz, 1H), 7.24 (s, 1H), 5.06-4.78 (m, 1H), 3.82 (s, 3H), 2.27-2.21 (m, 1H), 1.71-1.61 (m, 1H), 1.19-1.15 (m, 1H).
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (300.0 mg, 0.54 mmol) in 1,4-dioxane (12.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (276.5 mg, 2.68 mmol), BrettPhos (57.6 mg, 0.11 mmol), Cs2CO3 (524.3 mg, 1.61 mmol) and BrettPhos Pd G3 (48.6 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred with microwave at 120° C. for 1.5 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (100.0 mg, 30%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=626.3.
To a solution of (1S,2S)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (100.0 mg, 0.16 mmol) in DCM (4.0 mL) was added TFA (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (2.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 56% B in 7 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1H-indazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (12.3 mg, 21%) as a white solid. LCMS (ESI, m/z): [M+H]+=366.2. 1H NMR (300 MHz, DMSO-d6): δ 12.82 (s, 1H), 11.50 (s, 1H), 10.61 (s, 1H), 7.98-7.80 (m, 4H), 7.52 (d, J=2.7 Hz, 1H), 7.06 (s, 1H), 5.17-4.73 (m, 1H), 3.87 (s, 3H), 2.32-2.18 (m, 1H), 1.78-1.52 (m, 1H), 1.29-1.01 (m, 1H).
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (200.0 mg, 0.36 mmol) in 1,4-dioxane (6.0 mL) was added (1R,2R)-2-fluorocyclopropane-1-carboxamide (184.4 mg, 1.79 mmol), BrettPhos (38.4 mg, 0.07 mmol), Cs2CO3 (349.6 mg, 1.07 mmol) and BrettPhos Pd G3 (32.4 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was irradiated with microwave radiation at 120° C. for 1.5 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with ether/ethyl acetate (2/1, v/v) to afford (1R,2R)-2-fluoro-N-[3-(4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (92.0 mg, 41%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=626.3.
To a solution of (1R,2R)-2-fluoro-N-[3-(4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (40.0 mg, 0.06 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 20×250 mm, 5 um, 12 nm; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 22% B to 52% B in 7 min; 254 nm) to afford (1R,2R)-2-fluoro-N-[3-(4-methoxy-1H-1,3-benzodiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (5.2 mg, 22%) as a white solid. LCMS (ESI, m/z): [M+H]+=366.2. 1H NMR (300 MHz, DMSO-d6): δ 12.60 (s, 1H), 11.60 (s, 1H), 10.74 (s, 1H), 8.28 (s, 1H), 8.12-7.98 (m, 2H), 7.58 (s, 1H), 7.45 (d, J=8.4 Hz, 1H), 7.35 (d, J=8.1 Hz, 1H), 5.15-4.91 (m, 1H), 4.32 (s, 2H), 3.75 (s, 1H), 2.38-2.31 (m, 1H), 1.81-1.71 (m, 1H), 1.34-1.20 (m, 1H).
To a solution of 5-bromo-6-methoxy-1H-1,3-benzodiazole (1.0 g, 4.40 mmol) in THE (20.0 mL) was added NaH (0.2 g, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. Then SEM-Cl (0.9 g, 5.28 mmol) was added to the mixture. The resulting mixture was stirred at 0° C. for another 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/EtOAc (1/1, v/v) to afford 5-bromo-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (1.5 g, 95%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=357.1.
To a solution of 5-bromo-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (750.0 mg, 2.10 mmol) in 1,4-dioxane (20.0 mL) was added bis(pinacolato)diboron (639.6 mg, 2.52 mmol), KOAc (618.0 mg, 6.30 mmol) and Pd(dppf)Cl2 (153.6 mg, 0.21 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (6/1, v/v) to afford 6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (420.0 mg, 49%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=405.2.
To a solution of 6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (870.0 mg, 2.15 mmol) in 1,4-dioxane/H2O (15.0/3.0 mL) was added 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (778.2 mg, 2.15 mmol), K2CO3 (892.0 mg, 6.45 mmol) and Pd(dppf)Cl2 (314.8 mg, 0.43 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (590.0 mg, 49%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=559.2.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (540.0 mg, 0.97 mmol) in 1,4-dioxane (10.0 mL) was added (1R,2R)-2-fluorocyclopropane-1-carboxamide (497.7 mg, 4.83 mmol), Cs2CO3 (943.8 mg, 2.90 mmol), BrettPhos (103.7 mg, 0.19 mmol) and BrettPhos Pd G3 (87.5 mg, 0.10 mmol) at room temperature under N2. The reaction mixture was irradiated with microwave radiation at 100° C. for 2 h. After the reaction was completed, the mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1R,2R)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (180.0 mg, 29%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=626.3.
To a solution of (1R,2R)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (180.0 mg, 0.29 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 22% B to 52% B in 7 min; 254 nm) to afford (1R,2R)-2-fluoro-N-[3-(6-methoxy-1H-1,3-benzodiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (20.1 mg, 19%) as a white solid. LCMS (ESI, m/z): [M+H]+=366.1. 1H NMR (300 MHz, DMSO-d6): δ 12.25 (s, 1H), 11.50 (s, 1H), 10.62 (s, 1H), 8.11 (s, 1H), 7.97-7.88 (m, 2H), 7.72-7.57 (m, 1H), 7.53 (s, 1H), 7.41-7.10 (m, 1H), 5.03-4.81 (m, 1H), 3.83 (s, 3H), 2.29-2.18 (m, 1H), 1.71-1.60 (m, 1H), 1.19-1.12 (m, 1H).
To a solution of 5-bromo-6-methoxy-1H-indazole (3.0 g, 6.60 mmol) in THF (45.0 mL) was added NaH (475.6 mg, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. Then SEM-Cl (4.4 g, 26.43 mmol) was added dropwise to the mixture at 0° C. under N2. The resulting mixture was stirred at 0° C. for another 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/2, v/v) to afford 5-bromo-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (3.0 g, 64%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=357.1.
To a solution of 5-bromo-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (2.5 g, 6.99 mmol) in dioxane (40.0 mL) was added bis(pinacolato)diboron (2.6 g, 10.54 mmol), KOAc (2.0 g, 20.99 mmol) and Pd(dppf)Cl2 (511.9 mg, 0.70 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (1.1 g, 39%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=405.2.
To a solution of 6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (1.0 g, 2.47 mmol) in dioxane/H2O (10.0/1.0 mL) was added Pd(dppf)Cl2 (180.9 mg, 0.27 mmol), 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (894.5 mg, 2.47 mmol) and K2CO3 (1025.9 mg, 7.49 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (500.0 mg, 48%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=559.2.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (250.0 mg, 0.47 mmol) in dioxane (10.0 mL) was added Cs2CO3 (436.9 mg, 1.31 mmol), (1R,2R)-2-fluorocyclopropane-1-carboxamide (138.6 mg, 1.34 mmol), Brettphos Pd G3 (81.0 mg, 0.09 mmol) and BrettPhos (23.9 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1R,2R)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (100.0 mg, 36%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=626.3.
To a solution of (1R,2R)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (100.0 mg, 0.16 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 9% B to 26% B in 8 min; 254 nm) to afford (1R,2R)-2-fluoro-N-[3-(6-methoxy-1H-indazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (4.5 mg, 8%) as a white solid. LCMS (ESI, m/z): [M+H]+=366.1. 1H NMR (400 MHz, DMSO-d6): δ 12.82 (s, 1H), 11.49 (s, 1H), 10.60 (s, 1H), 7.97-7.94 (m, 2H), 7.91-7.85 (m, 1H), 7.80 (s, 1H), 7.51 (d, J=2.4 Hz, 1H), 7.06 (s, 1H), 5.01-4.83 (m, 1H), 3.86 (s, 3H), 2.29-2.18 (m, 1H), 1.69-1.62 (m, 1H), 1.18-1.15 (m, 1H).
A solution of 6-chloro-3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (300.0 mg, 0.77 mmol), (1S,2S)-2-fluorocyclopropane-1-carboxamide (396.6 mg, 3.85 mmol), Cs2CO3 (752.0 mg, 2.31 mmol), BrettPhos (82.6 mg, 0.15 mmol) and BrettPhos Pd G3 (69.7 mg, 0.08 mmol) in dioxane (10.0 mL) was irradiated with microwave radiation at 120° C. for 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (94/6, v/v) to afford (1S,2S)-2-fluoro-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (250.0 mg, 71%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=457.2.
A mixture of (1S,2S)-2-fluoro-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (250.0 mg, 0.55 mmol) and TFA (5.0 mL) in CH2Cl2 (5.0 mL) was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum. The residue was dissolved in NH3·H2O (5.0 mL) and ACN (5.0 mL). The mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with Water/ACN (54/46, v/v) and then purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 20×250 mm, 5 um, 12 nm; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 56% B in 10 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (82.4 mg, 46%) as a white solid. LCMS (ESI, m/z): [M+H]+=327.2 1H NMR (300 MHz, DMSO-d6): δ 13.45 (s, 1H), 10.94 (s, 1H), 8.11 (d, J=9.0 Hz, 1H), 7.98 (d, J=8.7 Hz, 1H), 7.64-7.60 (m, 1H), 7.47-7.41 (m, 1H), 7.19 (d, J=7.8 Hz, 1H), 7.09-7.04 (m, 1H), 5.07-4.83 (m, 1H), 3.83 (s, 3H), 2.29-2.24 (m, 1H), 1.72-1.63 (m, 1H), 1.23-1.11 (m, 1H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)imidazo[1,2-a]pyridine (337.4 mg, 1.38 mmol), K2CO3 (573.1 mg, 4.15 mmol) and Pd(dppf)Cl2 (101.1 mg, 0.07 mmol). The resulting mixture was stirred at 80° C. for 3 h under N2. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford 6-chloro-3-[imidazo[1,2-a]pyridin-7-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (470.0 mg, 85%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=399.1.
To a solution of 6-chloro-3-[imidazo[1,2-a]pyridin-7-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (440.0 mg, 1.10 mmol) in 1,4-dioxane (10.0 mL) was added cyclopropanecarboxamide (469.3 mg, 5.51 mmol), BrettPhos (118.4 mg, 0.22 mmol), Cs2CO3 (1.1 g, 3.31 mmol) and BrettPhos Pd G3 (100.0 mg, 0.11 mmol). The resulting mixture was stirred at 100° C. for 2 h under N2. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (95/5, v/v) to afford N-(3-[imidazo[1,2-a]pyridin-7-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (450.0 mg, 91%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=448.2.
To a solution of N-(3-[imidazo[1,2-a]pyridin-7-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (400.0 mg, 0.89 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL). The mixture was stirred at room temperature for 1 h. The mixture was concentrated under vacuum. The residue was dissolved in ACN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for additional 16 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 47% B in 10 min; 254 nm) to afford N-(3-[imidazo[1,2-a]pyridin-7-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (16.4 mg, 5%) as a white solid. LCMS (ESI, m/z): [M+H]+=318.2. 1H NMR (400 MHz, DMSO-d6): δ 11.83 (s, 1H), 10.66 (s, 1H), 8.55 (d, J=7.2 Hz, 1H), 8.40 (d, J=8.8 Hz, 1H), 8.00-7.99 (m, 2H), 7.89 (s, 1H), 7.82 (s, 1H), 7.53 (s, 1H), 7.35-7.33 (m, 1H), 2.08-2.03 (m, 1H), 0.86-0.81 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in dioxane/H2O (15.0/3.0 mL) was added 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)imidazo[1,2-a]pyridine (337.4 mg, 1.38 mmol), K2CO3 (573.9 mg, 4.17 mmol) and Pd(dppf)Cl2 (101.4 mg, 0.18 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (95/5, v/v) to afford 6-chloro-3-[imidazo[1,2-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (200.0 mg, 82%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=399.1.
To a solution of 6-chloro-3-[imidazo[1,2-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (430.0 mg, 1.08 mmol) in dioxane (10.0 mL) was added cyclopropanecarboxamide (275.8 mg, 3.23 mmol), BrettPhos (115.7 mg, 0.26 mmol), Brettphos Pd G3 (97.7 mg, 0.32 mmol) and Cs2CO3 (1053.8 mg, 3.23 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford N-(3-[imidazo[1,2-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (450.0 mg, 93%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=448.2.
To a solution of N-(3-[imidazo[1,2-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (400.0 mg, 0.89 mmol) in CH2Cl2 (10.0 mL) was added TFA (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (10.0 mL) and NH3·H2O (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 18% B to 48% B in 8 min; 254 nm) to afford N-(3-(imidazo[1,2-a]pyridin-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (21.7 mg, 7%) as a white solid. LCMS (ESI, m/z): [M+H]+=318.1. 1H NMR (300 MHz, DMSO-d6): δ 11.72 (s, 1H), 10.65 (s, 1H), 8.94 (s, 1H), 8.37 (d, J=8.7 Hz, 1H), 7.99-7.97 (m, 2H), 7.84 (d, J=2.7 Hz, 1H), 7.64-7.55 (m, 3H), 2.06-1.99 (m, 1H), 0.84-0.79 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-one (121.3 mg, 0.99 mmol), K2CO3 (389.9 mg, 1.65 mmol) and Pd(dppf)Cl2 (121.8 mg, 0.14 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (98/2, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-1-methylpyridin-2-one (360.0 mg, 67%) as a white solid. LCMS (ESI, m/z): [M+H]+=390.1.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-1-methylpyridin-2-one (310.0 mg, 0.79 mmol) in dioxane (4.0 mL) was added cyclopropanecarboxamide (338.3 mg, 3.97 mmol), BrettPhos (85.3 mg, 0.16 mmol), Cs2CO3 (777.0 mg, 2.38 mmol) and BrettPhos Pd G3 (72.0 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (69/31, v/v) to afford N-[3-(1-methyl-6-oxopyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (340.0 mg, 97%) as a white solid. LCMS (ESI, m/z): [M+H]+=439.2.
To a solution of N-[3-(1-methyl-6-oxopyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (300.0 mg, 0.68 mmol) in DCM (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in ACN (2.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 19% B to 49% B in 7 min; 254 nm) to afford N-[3-(1-methyl-6-oxopyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (71.6 mg, 34%) as a white solid. LCMS (ESI, m/z): [M+H]+=309.2. 1H NMR (400 MHz, DMSO-d6): δ 11.53 (s, 1H), 10.58 (s, 1H), 8.20 (d, J=8.8 Hz, 1H), 7.97 (d, J=2.4 Hz, 1H), 7.90 (d, J=8.8 Hz, 1H), 7.82-7.80 (m, 1H), 7.59 (d, J=2.0 Hz, 1H), 6.46 (d, J=9.6 Hz, 1H), 3.51 (s, 3H), 2.05-1.98 (m, 1H), 0.83-0.77 (m, 4H).
To a solution of 6-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (450.0 mg, 0.81 mmol) in 1,4-dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (414.8 mg, 4.02 mmol), BrettPhos (86.4 mg, 0.16 mmol), Cs2CO3 (786.5 mg, 2.41 mmol) and BrettPhos Pd G3 (72.9 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was irradiated with microwave radiation at 120° C. for 1.5 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (70.0 mg, 14%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=626.3.
To a solution of (1S,2S)-2-fluoro-N-[3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (70.0 mg, 0.11 mmol) in DCM (4.0 mL) was added TFA (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (2.0 mL) and NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (YMC-Actus Triart C18, 20×250 mm, 5 um, 12 nm; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 29% B to 59% B in 7 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(5-methoxy-1H-indazol-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (9.2 mg, 23%) as a white solid. LCMS (ESI, m/z): [M+H]+=366.2. 1H NMR (300 MHz, DMSO-d6): δ 12.81 (s, 1H), 11.60 (s, 1H), 10.62 (s, 1H), 8.02-7.92 (m, 3H), 7.63-7.61 (m, 2H), 7.30 (s, 1H), 5.14-4.82 (m, 1H), 3.83 (s, 3H), 2.29-2.16 (m, 1H), 1.79-1.52 (m, 1H), 1.34-1.05 (m, 1H).
To a solution of 6-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (250.0 mg, 0.45 mmol) in 1,4-dioxane (4.0 mL) was added (1R,2R)-2-fluorocyclopropane-1-carboxamide (230.4 mg, 2.24 mmol), BrettPhos (48.0 mg, 0.09 mmol), Cs2CO3 (349.6 mg, 1.07 mmol) and BrettPhos Pd G3 (40.5 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was irradiated with microwave radiation at 120° C. for 1.5 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with ether/ethyl acetate (3/2, v/v) to afford (1R,2R)-2-fluoro-N-[3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (200.0 mg, 71%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=626.3.
To a solution of (1R,2R)-2-fluoro-N-[3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (200.0 mg, 0.32 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 23% B to 53% B in 7 min; 254 nm) to afford (1R,2R)-2-fluoro-N-[3-(5-methoxy-1H-indazol-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (30.7 mg, 26%) as a white solid. LCMS (ESI, m/z): [M+H]+=366.2. 1H NMR (300 MHz, DMSO-d6): δ 12.78 (s, 1H), 11.57 (s, 1H), 10.60 (s, 1H), 7.96-7.84 (m, 3H), 7.57-7.54 (m, 2H), 7.24 (s, 1H), 5.00-4.73 (m, 1H), 3.76 (s, 3H), 2.20-2.15 (m, 1H), 1.66-1.54 (m, 1H), 1.20-1.00 (m, 1H).
To a solution of 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (400.0 mg, 0.98 mmol) in dioxane (15.0 mL) and H2O (1.0 mL) was added 6-bromo-5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[4,3-b]pyridine (140.4 mg, 0.78 mmol), Pd(dppf)Cl2 (35.8 mg, 0.08 mmol) and K2CO3 (202.8 mg, 2.98 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with dichloromethane/methyl alcohol (10/1, v/v) to afford 6-chloro-3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[4,3-b]pyridin-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (170.0 mg, 58%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=560.2.
To a solution of 6-chloro-3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[4,3-b]pyridin-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (120.0 mg, 0.24 mmol) in dioxane (5.0 mL) was added cyclopropanecarboxamide (54.9 mg, 0.63 mmol), Brettphos Pd G3 (19.4 mg, 0.01 mmol), BrettPhos (22.9 mg, 0.04 mmol) and Cs2CO3 (209.7 mg, 0.63 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford N-[3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[4,3-b]pyridin-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (100.0 mg, 60%) as a red oil. LCMS (ESI, m/z): [M+H]+=609.3.
To a solution of N-[3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[4,3-b]pyridin-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (100.0 mg, 0.16 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 56% B in 7 min; 254 nm) to afford N-(3-[5-methoxy-1H-pyrazolo[4,3-b]pyridin-6-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (2.4 mg, 4%) as an off-white solid. LCMS (ESI, m/z): [M+H]+=349.1. 1H NMR (300 MHz, Methanol-d4): δ 8.17-8.11 (m, 2H), 8.02 (s, 1H), 7.88 (d, J=8.7 Hz, 1H), 7.75 (s, 1H), 4.08 (s, 3H), 1.99-1.89 (m, 1H), 1.03-1.00 (m, 2H), 0.94-0.89 (m, 2H).
To a solution of 6-methoxy-3-nitropyridin-2-amine (10.0 g, 59.12 mmol) in CH2Cl2/MeOH (40.0 mL/10.0 mL) was added pyridinium Tribromide (28.3 g, 88.68 mmol). The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (70/30, v/v) to afford 5-bromo-6-methoxy-3-nitropyridin-2-amine (7.0 g, 48%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=248.0.
To a solution of 5-bromo-6-methoxy-3-nitropyridin-2-amine (5.0 g, 20.16 mmol) in HCOOH/i-PrOH (50.0 mL/50.0 mL) was added Fe (11.3 g, 201.58 mmol) and NH4Cl (10.8 g, 201.58 mmol). The resulting mixture was stirred at 80° C. for 5 h. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/90, v/v) to afford 6-bromo-5-methoxy-1H-imidazo[4,5-b]pyridine (3.0 g, 65%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=228.0.
To a solution of 6-bromo-5-methoxy-1H-imidazo[4,5-b]pyridine (3.0 g, 13.16 mmol) in THE (30.0 mL) was added sodium hydride (631.4 mg, 15.79 mmol) at 0° C. under N2. The mixture was stirred at 0° C. for 30 min. Then SEM-Cl (2.2 g, 13.16 mmol) was added to the mixture at 0° C. under N2. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the mixture was quenched with MeOH at 0° C. The resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (60/40, v/v) to afford 6-bromo-5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]imidazo[4,5-b]pyridine (1.4 g, 30%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=358.1.
To a solution of 6-bromo-5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]imidazo[4,5-b]pyridine (1.3 g, 3.77 mmol) in 1,4-dioxane (10.0 mL) was added bis(pinacolato)diboron (1.9 g, 7.54 mmol), KOAc (1.1 g, 11.30 mmol) and Pd(dppf)Cl2 (275.7 mg, 0.38 mmol). The mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by reverse phase flash chromatography with H2O/ACN (30/70, v/v) to afford 5-methoxy-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]imidazo[4,5-b]pyridine (820.0 mg, 53%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=406.2.
To a solution of 5-methoxy-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]imidazo[4,5-b]pyridine (220.0 mg, 0.54 mmol) in 1,4-dioxane/H2O (3.0 mL/1.0 mL) was added 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (196.32 mg, 0.54 mmol), K2CO3 (225.0 mg, 1.63 mmol) and Pd(dppf)Cl2 (39.7 mg, 0.05 mmol). The mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (12/88, v/v) to afford 6-chloro-3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]imidazo[4,5-b]pyridin-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (140.0 mg, 46%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=560.2.
To a solution of 6-chloro-3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]imidazo[4,5-b]pyridin-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (120.0 mg, 0.21 mmol) in 1,4-dioxane (3.0 mL) was added cyclopropanecarboxamide (54.7 mg, 0.64 mmol), Cs2CO3 (209.4 mg, 0.64 mmol), BrettPhos (23.0 mg, 0.043 mmol) and BrettPhos Pd G3 (19.4 mg, 0.02 mmol). The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (20/80, v/v) to afford N-[3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]imidazo[4,5-b]pyridin-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (100.0 mg, 76%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=609.3.
To a solution of N-[3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]imidazo[4,5-b]pyridin-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (80.0 mg, 0.13 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL). The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. The residue was dissolved in ACN (2.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18 Column, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 13% B to 43% B in 10 min; 254 nm) to afford N-(3-[5-methoxy-1H-imidazo[4,5-b]pyridin-6-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (14.6 mg, 32%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=349.2. 1H NMR (300 MHz, DMSO-d6): δ 12.32 (s, 1H), 11.60 (s, 1H), 10.59 (s, 1H), 8.22 (s, 1H), 8.08 (s, 1H), 8.01 (d, J=8.1 Hz, 1H), 7.92 (d, J=8.1 Hz, 1H), 7.63 (s, 1H), 3.96 (s, 3H), 2.09-2.01 (m, 1H), 0.86-0.79 (m, 4H).
To a solution of 3-bromo-6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (1.0 g, 2.76 mmol) in 1,4-dioxane/H2O (20.0/4.0 mL) was added (2-ethoxypyridin-3-yl)boronic acid (461.6 mg, 2.76 mmol), K2CO3 (1.1 g, 8.29 mmol) and Pd(dppf)Cl2 (202.3 mg, 0.28 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (96/4, v/v) to afford 6-chloro-3-(2-ethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (710.0 mg, 63%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=404.1.
To a solution of 6-chloro-3-(2-ethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (300.0 mg, 0.74 mmol) in 1,4-dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (382.8 mg, 3.71 mmol), Cs2CO3 (725.9 mg, 2.23 mmol), BrettPhos (79.7 mg, 0.15 mmol) and BrettPhos Pd G3 (67.3 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (70/30, v/v) to afford (1S,2S)—N-(3-(2-ethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (190.0 mg, 54%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=471.2.
To a solution of (1S,2S)—N-(3-(2-ethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (190.0 mg, 0.40 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 47% B in 10 min; 254 nm) to afford (1S,2S)—N-(3-(2-ethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (42.2 mg, 30%) as a white solid. LCMS (ESI, m/z): [M+H]+=341.2. 1H NMR (400 MHz, DMSO-d6): δ 11.69 (s, 1H), 10.66 (s, 1H), 8.15 (d, J=8.8 Hz, 1H), 8.06-7.92 (m, 3H), 7.75 (s, 1H), 7.07-7.04 (m, 1H), 5.01-4.84 (m, 1H), 4.44-4.39 (m, 2H), 2.29-2.22 (m, 1H), 1.69-1.63 (m, 1H), 1.37-1.34 (m, 3H), 1.20-1.13 (m, 1H).
To a solution of 6-chloro-3-(2-ethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (300.0 mg, 0.74 mmol) in 1,4-dioxane (10.0 mL) was added (1R,2R)-2-fluorocyclopropane-1-carboxamide (382.8 mg, 3.71 mmol), Cs2CO3 (725.9 mg, 2.23 mmol), BrettPhos (79.7 mg, 0.15 mmol) and BrettPhos Pd G3 (67.3 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford (1R,2R)—N-(3-(2-ethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (160.0 mg, 45%) as a white solid. LCMS (ESI, m/z): [M+H]+=471.2.
To a solution of (1R,2R)—N-(3-(2-ethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (110.0 mg, 0.23 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 20×250 mm, 5 um, 12 nm; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 30% B to 60% B in 10 min; 254 nm) to afford (1R,2R)—N-(3-(2-ethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (31.2 mg, 39%) as a white solid. LCMS (ESI, m/z): [M+H]+=341.2. 1H NMR (300 MHz, DMSO-d6): δ 11.70 (s, 1H), 10.68 (s, 1H), 8.15 (d, J=8.7 Hz, 1H), 8.07-8.05 (m, 1H), 8.01-7.98 (m, 1H), 7.95-7.92 (m, 1H), 7.75 (s, 1H), 7.08-7.04 (m, 1H), 5.06-4.81 (m, 1H), 4.45-4.38 (m, 2H), 2.26-2.22 (m, 1H), 1.70-1.59 (m, 1H), 1.38-1.33 (m, 3H), 1.20-1.13 (m, 1H).
A mixture of 6-chloro-3-iodo-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (1.4 g, 3.42 mmol), 2-ethoxypyridin-3-ylboronic acid (0.6 g, 3.42 mmol), K2CO3 (1.4 g, 10.25 mmol) and Pd(dppf)Cl2 (0.3 g, 0.34 mmol) in dioxane (20.0 mL) and H2O (4.0 mL) was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (92/8, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-3-yl)-2-ethoxypyridine (960.0 mg, 69%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=405.1.
A mixture of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-3-yl)-2-ethoxypyridine (300.0 mg, 0.74 mmol), (1S,2S)-2-fluorocyclopropane-1-carboxamide (381.9 mg, 3.70 mmol), K2CO3 (307.2 mg, 2.22 mmol), BrettPhos (79.5 mg, 0.15 mmol) and BrettPhos Pd G3 (67.15 mg, 0.07 mmol) in dioxane (15.0 mL) was irradiated with microwave radiation (MW) at 120° C. for 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (94/6, v/v) to afford (1S,2S)—N-[3-(2-ethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (330.0 mg, 94%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=472.2.
A mixture of (1S,2S)—N-[3-(2-ethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (330.0 mg, 0.70 mmol) and TFA (5.0 mL) in CH2Cl2 (5.0 mL) was stirred at room temperature for 16. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (94/6, v/v) and then purified by Prep-HPLC with the following conditions (Column: XBridge Shield RP18 OBD Column, 19×250 mm, 10 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: MeOH—Preparative; Flow rate: 25 mL/min; Gradient: 50% B to 58% B in 10 min; 220/254 nm) to afford (1S,2S)—N-[3-(2-ethoxypyridin-3-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (3.7 mg, 1%) as a white solid. LCMS (ESI, m/z): [M+H]+=342.2. 1H NMR (300 MHz, DMSO-d6): δ 13.59 (s, 1H), 10.97 (s, 1H), 8.28-8.23 (m, 2H), 8.06-8.00 (m, 2H), 7.14-7.10 (m, 1H), 5.07-4.83 (m, 1H), 4.48-4.41 (m, 2H), 2.30-2.25 (m, 1H), 1.73-1.63 (m, 1H), 1.32-1.21 (m, 4H).
To a solution of 3-bromopyridin-2-amine (5.0 g, 28.90 mmol) in THF (40.0 mL) was added NaH (5.8 g, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. Then CH3I (8.2 g, 57.80 mmol) was added dropwise to the mixture at 0° C. under N2. The resulting mixture was stirred at 0° C. for another 1 h. After the reaction was completed, the reaction mixture was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (4/1, v/v) to afford 3-bromo-N,N-dimethylpyridin-2-amine (4.6 g, 79%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=201.0.
To a solution of 3-bromo-N,N-dimethylpyridin-2-amine (123.0 mg, 0.61 mmol) in dioxane/H2O (16.0/4.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (250.0 mg, 0.61 mmol), K2CO3 (253.6 mg, 1.84 mmol) and Pd(dppf)Cl2 (44.8 mg, 0.06 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford 3-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-N,N-dimethylpyridin-2-amine (150.0 mg, 60%) as a brown oil. LCMS (ESI, m/z): [M+H]+=403.2.
To a solution of 3-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-N,N-dimethylpyridin-2-amine (130.0 mg, 0.32 mmol) in dioxane (4.0 mL) was added cyclopropanecarboxamide (164.7 mg, 1.94 mmol), Cs2CO3 (315.3 mg, 0.97 mmol), Brettphos (34.6 mg, 0.07 mmol) and BrettPhos Pd G3 (29.2 mg, 0.03 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford N-(3-(2-(dimethylamino)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (120.0 mg, 82%) as a white solid. LCMS (ESI, m/z): [M+H]+=452.2.
To a solution of N-(3-(2-(dimethylamino)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (100.0 mg, 0.22 mmol) in DCM (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in ACN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 2% to 21% in 7 min; 254 nm) to afford N-(3-(2-(dimethylamino)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (32.0 mg, 44%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=322.1. 1H NMR (400 MHz, CD3OD): δ 8.09-8.08 (m, 1H), 7.90 (d, J=8.8 Hz, 1H), 7.78 (d, J=8.4 Hz, 1H), 7.71-7.68 (m, 1H), 7.47 (s, 1H), 6.94-6.91 (m, 1H), 2.72 (s, 6H), 1.91-1.89 (m, 1H), 1.02-0.99 (m, 2H), 0.92-0.88 (m, 2H).
A mixture of 6-chloro-3-iodo-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (800.0 mg, 1.95 mmol), 2-fluoro-6-methoxyphenylboronic acid (331.8 mg, 1.95 mmol), K2CO3 (809.5 mg, 5.86 mmol) and Pd(dppf)Cl2 (142.9 mg, 0.20 mmol) in dioxane (20.0 mL) and H2O (2.0 mL) was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (95/5, v/v) to afford 6-chloro-3-(2-fluoro-6-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (500.0 mg, 63%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=408.1.
A mixture of 6-chloro-3-(2-fluoro-6-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (500.0 mg, 1.23 mmol), (1S,2S)-2-fluorocyclopropane-1-carboxamide (631.8 mg, 6.13 mmol), K2CO3 (508.2 mg, 3.68 mmol), BrettPhos (131.6 mg, 0.25 mmol) and BrettPhos Pd G3 (111.1 mg, 0.12 mmol) in dioxane (12.0 mL) was irradiated with microwave radiation (MW) at 120° C. for 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (94/6, v/v) to afford (1S,2S)-2-fluoro-N-[3-(2-fluoro-6-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (254.0 mg, 44%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=475.2.
A mixture of (1S,2S)-2-fluoro-N-[3-(2-fluoro-6-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (200.0 mg, 0.42 mmol) and TFA (5.0 mL) in CH2Cl2 (5.0 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in NH3·H2O (5.0 mL) and CAN (5.0 mL). The resulting mixture was stirred at room temperature for additional 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (94/6, v/v) and then purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 39% B in 10 min; 254/220 nm) to afford (1S,2S)-2-fluoro-N-[3-(2-fluoro-6-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (24.1 mg, 17%). LCMS (ESI, m/z): [M+H]+=345.1. 1H NMR (300 MHz, DMSO-d6): δ 13.54 (s, 1H), 10.97 (s, 1H), 8.00-7.89 (m, 2H), 7.50-7.47 (m, 1H), 7.05-6.93 (m, 2H), 5.05-4.81 (m, 1H), 3.76 (s, 3H), 2.29-2.24 (m, 1H), 1.77-1.63 (m, 1H), 1.22-1.17 (m, 1H).
To a solution of 6-chloro-3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (500.0 mg, 1.22 mmol) in 1,4-dioxane/H2O (10.0/2.0 mL) was added (2-ethoxy-6-fluorophenyl)boronic acid (224.5 mg, 1.22 mmol), K2CO3 (505.9 mg, 3.66 mmol) and Pd(dppf)Cl2 (89.2 mg, 0.12 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (75/25, v/v) to afford 6-chloro-3-(2-ethoxy-6-fluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (220.0 mg, 42%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=422.1.
To a solution of 6-chloro-3-(2-ethoxy-6-fluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (200.0 mg, 0.47 mmol) in 1,4-dioxane (6.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (244.3 mg, 2.37 mmol), K2CO3 (196.5 mg, 1.42 mmol), BrettPhos (50.9 mg, 0.10 mmol) and BrettPhos Pd G3 (43.0 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (75/25, v/v) to afford (1S,2S)—N-(3-(2-ethoxy-6-fluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (140.0 mg, 60%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=489.2.
To a solution of (1S,2S)—N-(3-(2-ethoxy-6-fluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (140.0 mg, 0.29 mmol) in DCM (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in ACN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 30% B to 60% B in 10 min; 254 nm) to afford (1S,2S)—N-(3-(2-ethoxy-6-fluorophenyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (26.4 mg, 25%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=359.1. 1H NMR (400 MHz, DMSO-d6): δ 13.55 (s, 1H), 11.00 (s, 1H), 8.00-7.95 (m, 2H), 7.49-7.43 (m, 1H), 7.03-6.92 (m, 2H), 5.06-4.85 (m, 1H), 4.11-4.05 (m, 2H), 2.29-2.25 (m, 1H), 1.71-1.63 (m, 1H), 1.23-1.12 (m, 4H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (1.0 g, 2.78 mmol) in THE (10.0 mL) was added LDA (4.4 mL, 2 mol/L) at −70° C. under N2. The mixture was stirred at −70° C. for 2 h. Then a solution of NFSI (2.3 g, 7.14 mmol) in THF (8.0 mL) was added dropwise to the mixture at −70° C. The mixture was stirred at −70° C. for another 3 h. After the reaction was completed, the mixture was quenched with aq·NH4Cl and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/CH2Cl2 (76/24, v/v) to afford 3-bromo-6-chloro-2-fluoro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (550.0 mg, 80%) as a white solid. LCMS (ESI, m/z): [M+H]+=379.0.
To a solution of 3-bromo-6-chloro-2-fluoro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (530.0 mg, 1.40 mmol) in 1,4-dioxane/H2O (5.0 mL/1.0 mL) was added 2-methoxypyridin-3-ylboronic acid (213.5 mg, 1.40 mmol), K2CO3 (578.7 mg, 4.19 mmol) and Pd(dppf)Cl2 (102.1 mg, 0.14 mmol). The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford 3-(6-chloro-2-fluoro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-methoxypyridine (300.0 mg, 52%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=408.1.
To a solution of 3-(6-chloro-2-fluoro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-methoxypyridine (240.0 mg, 0.59 mmol) in 1,4-dioxane (3.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (182.0 mg, 1.77 mmol), Cs2CO3 (575.1 mg, 1.77 mmol), BrettPhos (63.2 mg, 0.12 mmol) and BrettPhos Pd G3 (53.3 mg, 0.06 mmol). The resulting mixture was stirred at 100° C. for 2 h under N2. After the reaction was completed, the mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (60/40, v/v) to afford (1S,2S)-2-fluoro-N-[2-fluoro-3-(2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (100.0 mg, 35%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=475.2.
To a solution of (1S,2S)-2-fluoro-N-[2-fluoro-3-(2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (80.0 mg, 0.17 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL). The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. The residue was dissolved in ACN (2.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 32% B to 42% B in 8 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[2-fluoro-3-(2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (23.8 mg, 41%) as a white solid. LCMS (ESI, m/z): [M+H]+=345.1. 1H NMR (400 MHz, DMSO-d6): δ 12.55 (s, 1H), 10.64 (s, 1H), 8.18-8.16 (m, 1H), 7.91 (d, J=8.4 Hz, 1H), 7.82-7.77 (m, 2H), 7.11-7.08 (m, 1H), 5.02-4.82 (m, 1H), 3.90 (s, 3H), 2.24-2.20 (m, 1H), 1.69-1.60 (m, 1H), 1.18-1.12 (m, 1H).
To a solution of 6-chloro-1H-pyrrolo[2,3-b]pyridine (600.0 mg, 3.94 mmol) in THE (5.0 mL) was added NaH (133.6 mg, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. Then SEM-Cl (786.3 mg, 4.72 mmol) was added to the mixture at 0° C. under N2. The resulting mixture was stirred at 0° C. for another 1 h under N2. After the reaction was completed, the reaction was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (8/1, v/v) to afford 6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (440.0 mg, 39%) as a white solid. LCMS (ESI, m/z): [M+H]+=283.1.
To a solution of 6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (440.0 mg, 1.56 mmol) in dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (803.5 mg, 7.80 mmol), Cs2CO3 (1.5 g, 4.68 mmol), BrettPhos (167.5 mg, 0.31 mmol) and BrettPhos Pd G3 (141.5 mg, 0.15 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (65/35, v/v) to afford (1S,2S)-2-fluoro-N-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (170.0 mg, 31%) as a white solid. LCMS (ESI, m/z): [M+H]+=350.2.
To a solution of (1S,2S)-2-fluoro-N-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (170.0 mg, 0.48 mmol) in DMF (5.0 mL) was added Br2 (93.0 mg, 0.58 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the mixture was quenched with sat. NaHSO3 at 0° C. The resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (60/40, v/v) to afford (1S,2S)—N-(3-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (70.0 mg, 34%) as a white solid. LCMS (ESI, m/z): [M+H]+=428.1.
To a solution of (1S,2S)—N-(3-bromo-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (70.0 mg, 0.16 mmol) in dioxane/H2O (5.0/1.0 mL) was added 2-cyclopropoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (51.3 mg, 0.19 mmol), K2CO3 (66.2 mg, 0.48 mmol) and Pd(dppf)Cl2 (11.7 mg, 0.02 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford (1S,2S)—N-[3-(2-cyclopropoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (28.0 mg, 29%) as a white solid. LCMS (ESI, m/z): [M+H]+=483.2.
To a solution of (1S,2S)—N-[3-(2-cyclopropoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (28.0 mg, 0.06 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in ACN (3.0 mL) was added NH3·H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 32% B to 47% B in 7 min; 254 nm) to afford (1S,2S)—N-[3-(2-cyclopropoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (7.2 mg, 35%) as a white solid. LCMS (ESI, m/z): [M+H]+=353.2. 1H NMR (400 MHz, DMSO-d6): δ 11.69 (s, 1H), 10.66 (s, 1H), 8.12-8.05 (m, 2H), 7.99-7.86 (m, 2H), 7.64 (d, J=2.4 Hz, 1H), 7.12-7.09 (m, 1H), 5.03-4.82 (m, 1H), 4.38-4.34 (m, 1H), 2.27-2.20 (m, 1H), 1.70-1.61 (m, 1H), 1.21-1.11 (m, 1H), 0.82-0.74 (m, 2H), 0.71-0.67 (m, 2H).
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-(difluoromethoxy)pyridine (140.0 mg, 0.33 mmol) in dioxane (3.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (169.4 mg, 1.64 mmol), BrettPhos (35.3 mg, 0.06 mmol), Cs2CO3 (321.3 mg, 0.99 mmol) and BrettPhos Pd G3 (29.8 mg, 0.03 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford (1S,2S)—N-[3-[2-(difluoromethoxy)pyridin-3-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (45.0 mg, 28%) as a white solid. LCMS (ESI, m/z): [M+H]+=493.2.
To a solution of (1S,2S)—N-[3-[2-(difluoromethoxy)pyridin-3-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (45.0 mg, 0.09 mmol) in CH2Cl2 (1.0 mL) was added TFA (1.0 mL). The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (1.0 mL) and NH3·H2O (1.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 31% B to 61% B in 10 min; 254 nm) to afford (1S,2S)—N-[3-[2-(difluoromethoxy)pyridin-3-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (6.4 mg, 19%) as a white solid. LCMS (ESI, m/z): [M+H]+=363.1. 1H NMR (400 MHz, DMSO-d6): δ 11.82 (s, 1H), 10.66 (s, 1H), 8.15-8.09 (m, 3H), 7.97-7.61 (m, 3H), 7.36-7.32 (m, 1H), 5.00-4.81 (m, 1H), 2.24-2.20 (m, 1H), 1.68-1.60 (m, 1H), 1.17-1.14 (m, 1H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.83 mmol) in 1,4-dioxane/H2O (10.0/2.0 mL) was added 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)imidazo[1,2-a]pyridine (202.4 mg, 0.83 mmol), K2CO3 (343.9 mg, 2.49 mmol) and Pd(dppf)Cl2 (60.7 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) to afford 6-chloro-3-[imidazo[1,2-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (270.0 mg, 81%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=399.1.
To a solution of 6-chloro-3-[imidazo[1,2-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (250.0 mg, 0.63 mmol) in 1,4-dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (323.0 mg, 3.13 mmol), BrettPhos (67.3 mg, 0.13 mmol), Cs2CO3 (612.5 mg, 1.88 mmol) and BrettPhos Pd G3 (56.8 mg, 0.06 mmol) at room temperature under N2. The resulting mixture was stirred with microwave at 120° C. for 90 min. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) to afford (1S,2S)-2-fluoro-N-(3-[imidazo[1,2-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (160.0 mg, 55%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=466.2.
To a solution of (1S,2S)-2-fluoro-N-(3-[imidazo[1,2-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (160.0 mg, 0.34 mmol) in DCM (6.0 mL) was added TFA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. the residue was dissolved in CH3CN (6.0 mL) and NH3·H2O (6.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 47% B in 10 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-[imidazo[1,2-a]pyridin-6-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (5.5 mg, 5%) as a white solid. LCMS (ESI, m/z): [M+H]+=336.2. 1H NMR (300 MHz, DMSO-d6): δ 11.73 (s, 1H), 10.58 (s, 1H), 8.94 (s, 1H), 8.39 (d, J=8.7 Hz, 1H), 7.98 (d, J=6.0 Hz, 2H), 7.86 (s, 1H), 7.67-7.56 (m, 3H), 5.05-4.82 (m, 1H), 2.30-2.21 (m, 1H), 1.71-1.63 (m, 1H), 1.22-1.13 (m, 1H).
To a solution of 5-bromo-4-methoxypyridin-2-amine (1.0 g, 4.93 mmol) in CH2Cl2 (5.0 mL) was added 2-chloroacetaldehyde (2.1 g, 10.84 mmol) and saturated NaHCO3 solution (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the reaction was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) to afford 6-bromo-7-methoxyimidazo[1,2-a]pyridine (380.0 mg, 33%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=227.0.
To a solution of 6-bromo-7-methoxyimidazo[1,2-a]pyridine (140.0 mg, 0.62 mmol) in dioxane/H2O (16.0/4.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (252.0 mg, 0.62 mmol), K2CO3 (255.6 mg, 1.85 mmol) and Pd(dppf)Cl2 (45.1 mg, 0.06 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) to afford 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-7-methoxyimidazo[1,2-a]pyridine (200.0 mg, 75%) as a brown oil. LCMS (ESI, m/z): [M+H]+=429.1.
To a solution of 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-7-methoxyimidazo[1,2-a]pyridine (180.0 mg, 0.42 mmol) in dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (216.3 mg, 2.10 mmol), Cs2CO3 (401.1 mg, 1.26 mmol), Brettphos (45.0 mg, 0.08 mmol) and BrettPhos Pd G3 (38.0 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) to afford (1S,2S)-2-fluoro-N-(3-(7-methoxyimidazo[1,2-a]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (160.0 mg, 76%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=496.2.
To a solution of (1S,2S)-2-fluoro-N-(3-(7-methoxyimidazo[1,2-a]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (140.0 mg, 0.28 mmol) in DCM (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in ACN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with MeOH/H2O (56/44, v/v) to afford to afford (1S,2S)-2-fluoro-N-(3-(7-methoxyimidazo[1,2-a]pyridin-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (9.0 mg, 8%) as a white solid. LCMS (ESI, m/z): [M+H]+=366.3. 1H NMR (400 MHz, DMSO-d6): δ 10.64 (s, 1H), 8.61 (s, 1H), 8.04 (d, J=8.8 Hz, 1H), 7.90 (d, J=8.8 Hz, 1H), 7.78 (s, 1H), 7.60 (s, 1H), 7.40-7.37 (m, 1H), 7.03 (s, 1H), 5.02-4.81 (m, 1H), 3.87 (s, 3H), 2.25-2.19 (m, 1H), 1.70-1.61 (m, 1H), 1.35-1.04 (m, 1H).
To a solution of 5-bromo-6-chloropyridin-2-amine (6.0 g, 28.92 mmol) in EtOH/H2O (48.0/12.0 mL) was added NaHCO3 (2.9 g, 34.71 mmol) and 2-chloroacetaldehyde (6.8 g, 34.71 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (97/3, v/v) to afford 6-bromo-5-chloroimidazo[1,2-a]pyridine (2.4 g, 35%) as a brown solid. LCMS (ESI, m/z): [M+H]+=230.9.
To a solution of 6-bromo-5-chloroimidazo[1,2-a]pyridine (1.0 g, 4.32 mmol) in THE (15.0 mL) was added CH3OH (207.6 mg, 6.48 mmol) and NaH (345.6 mg, 60%) at room temperature under N2. The resulting mixture was stirred at 60° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (96/4, v/v) to afford 6-bromo-5-methoxyimidazo[1,2-a]pyridine (550.0 mg, 56%) as a brown solid. LCMS (ESI, m/z): [M+H]+=227.0.
To a solution of 6-bromo-5-methoxyimidazo[1,2-a]pyridine (155.0 mg, 0.68 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (279.1 mg, 0.68 mmol), K2CO3 (283.0 mg, 2.05 mmol) and Pd(dppf)Cl2 (50.0 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to afford 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-methoxyimidazo[1,2-a]pyridine (162.0 mg, 52%) as a brown solid. LCMS (ESI, m/z): [M+H]+=429.1.
To a solution of 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-methoxyimidazo[1,2-a]pyridine (160.0 mg, 0.37 mmol) in 1,4-dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (192.3 mg, 1.87 mmol), Cs2CO3 (364.6 mg, 1.12 mmol), BrettPhos (40.0 mg, 0.08 mmol) and BrettPhos Pd G3 (33.8 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (90/10, v/v) to afford (1S,2S)-2-fluoro-N-(3-(5-methoxyimidazo[1,2-a]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (140.0 mg, 47%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=496.2.
To a solution of (1S,2S)-2-fluoro-N-(3-(5-methoxyimidazo[1,2-a]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (140.0 mg, 0.28 mmol) in DCM (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in ACN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 21% B to 30% B in 8 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-(5-methoxyimidazo[1,2-a]pyridin-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl) cyclopropane-1-carboxamide (6.3 mg, 6%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=366.2. 1H NMR (300 MHz, DMSO-d6): δ 11.33 (s, 1H), 10.61 (s, 1H), 8.26 (d, J=8.7 Hz, 1H), 8.02-7.97 (m, 2H), 7.92-7.84 (m, 1H), 7.79 (d, J=2.1 Hz, 1H), 7.62 (d, J=2.1 Hz, 1H), 6.38 (d, J=8.4 Hz, 1H), 5.11-4.78 (m, 1H), 3.75 (s, 3H), 2.23-2.13 (m, 1H), 1.78-1.53 (m, 1H), 1.24-1.08 (m, 1H).
To a solution of 3-bromo-5-fluoro-2-methoxypyridine (1.0 g, 4.85 mmol) in dioxane (30.0 mL) was added bis(pinacolato)diboron (3.7 g, 14.56 mmol), KOAc (1.43 g, 14.56 mmol) and Pd(dppf)Cl2 (0.4 g, 0.49 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with H2O/ACN (2/1, v/v) to afford 5-fluoro-2-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (600.0 mg, 49%) as a white solid. LCMS (ESI, m/z): [M+H]+=254.1.
To a solution of 5-fluoro-2-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (556.0 mg, 2.20 mmol) in dioxane/H2O (10.0 mL/2.0 mL) was added 6-chloro-3-iodo-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (900.2 mg, 2.20 mmol), K2CO3 (910.9 mg, 6.59 mmol) and Pd(dppf)Cl2 (160.8 mg, 0.22 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-3-yl)-5-fluoro-2-methoxypyridine (890.0 mg, 99%) as a white solid. LCMS (ESI, m/z): [M+H]+=409.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-3-yl)-5-fluoro-2-methoxypyridine (170.0 mg, 0.42 mmol) in 1,4-dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (214.3 mg, 2.08 mmol), BrettPhos (44.63 mg, 0.08 mmol), Cs2CO3 (406.4 mg, 1.25 mmol) and BrettPhos Pd G3 (37.7 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(5-fluoro-2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (40.0 mg, 20%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=476.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(5-fluoro-2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (120.0 mg, 0.25 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 27% B to 51% B in 7 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(5-fluoro-2-methoxypyridin-3-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (19.1 mg, 22%) as a white solid. LCMS (ESI, m/z): [M+H]+=346.2. 1H NMR (400 MHz, DMSO-d6): δ 13.74 (s, 1H), 10.99 (s, 1H), 8.27-8.24 (m, 2H), 8.04-7.97 (m, 2H), 5.05-4.85 (m, 1H), 3.96 (s, 3H), 2.30-2.23 (m, 1H), 1.72-1.65 (m, 1H), 1.25-1.12 (m, 1H).
To a solution of 3-nitropyridine-2,4-diol (5.0 g, 32.03 mmol) in CHCl3 (50.0 mL) was added CH3I (30.5 g, 256.21 mmol) and Ag2CO3 (35.3 g, 128.12 mmol). The resulting mixture was stirred at room temperature for 12 h. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (63/37, v/v) to afford 2,4-dimethoxy-3-nitropyridine (3.8 g, 64%) as an off-white solid. LCMS (ESI, m/z): [M+H]+=185.0.
To a solution of 2,4-dimethoxy-3-nitropyridine (3.7 g, 20.92 mmol) in THE (50.0 mL) was added Pd/C (1.1 g, dry) at room temperature. The resulting mixture was stirred at room temperature for 16 h under H2. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure to afford 2,4-dimethoxypyridin-3-amine (3.0 g, crude) as an off-white solid. LCMS (ESI, m/z): [M+H]+=155.1.
To a solution of 2,4-dimethoxypyridin-3-amine (1.4 g, 9.08 mmol) in CH3CN (30.0 mL) was added LiBr (2.7 g, 27.23 mmol), t-BuONO (1.7 g, 16.36 mmol) and CuBr2 (1.6 g, 10.89 mmol). The resulting mixture was stirred at 60° C. for 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with H2O/ACN (36/64, v/v) to afford 3-bromo-2,4-dimethoxypyridine (1.0 g, 51%) as a red solid. LCMS (ESI, m/z): [M+H]+=218.0.
To a solution of 3-bromo-2,4-dimethoxypyridine (500.0 mg, 2.29 mmol) in H2O (1.0 mL) and dioxane (10.0 mL) was added K2CO3 (950.4 mg, 6.89 mmol), 6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-ylboronic acid (898.8 mg, 2.75 mmol) and Pd(dppf)Cl2 (167.7 mg, 0.29 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2,4-dimethoxypyridine (230.0 mg, 24%) as a red oil. LCMS (ESI, m/z): [M+H]+=420.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2,4-dimethoxypyridine (200.0 mg, 0.47 mmol) in dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (245.8 mg, 2.31 mmol), Brettphos Pd G3 (43.7 mg, 0.08 mmol), BrettPhos (51.2 mg, 0.09 mmol) and Cs2CO3 (465.8 mg, 1.49 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 3 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)—N-[3-(2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (110.0 mg, 48%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=487.2.
To a solution of (1S,2S)—N-[3-(2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (100.0 mg, 0.19 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 30% B to 39% B in 9 min; 254 nm) to afford (1S,2S)—N-[3-(2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (28.2 mg, 43%) as a white solid. LCMS (ESI, m/z): [M+H]+=357.2. 1H NMR (300 MHz, DMSO-d6): δ 11.53 (s, 1H), 10.58 (s, 1H), 8.06 (d, J=6.0 Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 7.61 (d, J=8.7 Hz, 1H), 7.38 (d, J=2.4 Hz, 1H), 6.88 (d, J=5.7 Hz, 1H), 5.04-4.80 (m, 1H), 3.81 (s, 3H), 3.79 (s, 3H), 2.27-2.18 (m, 1H), 1.72-1.61 (m, 1H), 1.28-1.08 (m, 1H).
To a solution of 5-fluoro-2-methoxypyridin-4-ol (3.0 g, 20.96 mmol) in ACN (125.0 mL) was added NBS (3.5 g, 19.91 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was dissolved in CHCl3 (250.0 mL). Then Ag2CO3 (15.4 g, 55.85 mmol) and CH3I (7.9 g, 55.87 mmol) were added to the mixture at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 3-bromo-5-fluoro-2,4-dimethoxypyridine (3.1 g, 94%) as a white solid. LCMS (ESI, m/z): [M+H]+=236.0.
To a solution of 3-bromo-5-fluoro-2,4-dimethoxypyridine (173.0 mg, 0.73 mmol) in dioxane/H2O (10.0 mL/2.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (299.6 mg, 0.73 mmol), K2CO3 (303.8 mg, 2.20 mmol) and Pd(dppf)Cl2 (53.6 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-fluoro-2,4-dimethoxypyridine (220.0 mg, 68%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=438.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-fluoro-2,4-dimethoxypyridine (200.0 mg, 0.46 mmol) in 1,4-dioxane (15.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (235.4 mg, 2.28 mmol), BrettPhos (49.0 mg, 0.09 mmol), Cs2CO3 (446.4 mg, 1.37 mmol) and BrettPhos Pd G3 (41.4 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(5-fluoro-2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (90.0 mg, 39%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=505.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(5-fluoro-2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (90.0 mg, 0.18 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the reaction was diluted with H2 and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 38% B to 47% B in 8 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(5-fluoro-2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (19.3 mg, 29%) as a white solid. LCMS (ESI, m/z): [M+H]+=375.2. 1H NMR (400 MHz, DMSO-d6): δ 11.70 (s, 1H), 10.63 (s, 1H), 8.12 (d, J=3.2 Hz, 1H), 7.87 (d, J=8.4 Hz, 1H), 7.71 (d, J=8.8 Hz, 1H), 7.47 (d, J=2.4 Hz, 1H), 5.02-4.82 (m, 1H), 3.81-3.80 (m, 6H), 2.25-2.22 (m, 1H), 1.70-1.63 (m, 1H), 1.25-1.04 (m, 1H).
To a solution of 6-chloro-3-iodo-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (300.3 mg, 0.73 mmol) in dioxane/H2O (10.0 mL/2.0 mL) was added 2-methoxypyridin-3-ylboronic acid (112.0 mg, 0.73 mmol), K2CO3 (303.6 mg, 2.20 mmol) and Pd(dppf)Cl2 (53.6 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-3-yl)-2-methoxypyridine (227.0 mg, 79%) as a white solid. LCMS (ESI, m/z): [M+H]+=391.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-3-yl)-2-methoxypyridine (170.0 mg, 0.44 mmol) in 1,4-dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (224.2 mg, 2.17 mmol), BrettPhos (46.7 mg, 0.09 mmol), Cs2CO3 (425.1 mg, 1.31 mmol) and BrettPhos Pd G3 (39.4 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (138.0 mg, 70%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=458.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (138.0 mg, 0.30 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 29% B to 59% B in 10 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(2-methoxypyridin-3-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (17.3 mg, 18%) as a white solid. LCMS (ESI, m/z): [M+H]+=328.2. 1H NMR (300 MHz, DMSO-d6): δ 13.60 (s, 1H), 10.96 (s, 1H), 8.29-8.20 (m, 2H), 8.08-8.00 (m, 2H), 7.16-7.12 (m, 1H), 5.07-4.84 (m, 1H), 3.96 (s, 3H), 2.29-2.25 (m, 1H), 1.73-1.60 (m, 1H), 1.23-1.18 (m, 1H).
To a solution of 6-chloro-3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (562.9 mg, 1.37 mmol) in dioxane/H2O (16.0/4.0 mL) was added (2,6-dimethoxyphenyl)boronic acid (250.0 mg, 1.37 mmol), K2CO3 (569.6 mg, 4.12 mmol) and Pd(dppf)Cl2 (100.5 mg, 0.14 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 6-chloro-3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (200.0 mg, 34%) as a brown oil. LCMS (ESI, m/z): [M+H]+=420.1.
To a solution of 6-chloro-3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (180.0 mg, 0.43 mmol) in dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (220.9 mg, 2.14 mmol), Cs2CO3 (418.9 mg, 1.29 mmol), Brettphos (46.0 mg, 0.09 mmol) and BrettPhos Pd G3 (38.9 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (10/1, v/v) to afford (1S,2S)—N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (60.0 mg, 19%) as a white solid. LCMS (ESI, m/z): [M+H]+=487.2.
To a solution of (1S,2S)—N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (100.0 mg, 0.21 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in ACN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 21% to 51% in 7 min; 254 nm) to afford (1S,2S)—N-(3-(2,6-dimethoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (22.1 mg, 30%) as a white solid. LCMS (ESI, m/z): [M+H]+=357.2. 1H NMR (400 MHz, DMSO-d6): δ 13.31 (s, 1H), 10.92 (s, 1H), 7.92-7.90 (m, 1H), 7.76-7.74 (m, 1H), 7.43-7.39 (m, 1H), 6.80-6.78 (m, 2H), 5.07-4.84 (m, 1H), 3.66 (s, 6H), 2.32-2.21 (m, 1H), 1.75-1.61 (m, 1H), 1.22-1.13 (m, 1H).
To a solution of 6-chloro-3-iodo-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (1.5 g, 3.66 mmol) in dioxane/H2O (5.0/1.0 mL) was added 5-fluoro-2-methoxyphenylboronic acid (750.0 mg, 4.39 mmol), K2CO3 (1.5 g, 10.9 mmol) and Pd(dppf)Cl2 (270.0 mg, 0.36 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to afford 6-chloro-3-(5-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (1.0 g, 67%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=408.1.
To a solution of 6-chloro-3-(5-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (980.0 mg, 2.44 mmol) in dioxane (5.0 mL) was added diphenylmethanimine (1.3 g, 7.21 mmol), BrettPhos (257.9 mg, 0.48 mmol), Cs2CO3 (2.3 g, 7.21 mmol) and BrettPhos Pd G3 (217.7 mg, 0.24 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (85/15, v/v) to afford N-[3-(5-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]-1,1-diphenylmethanimine (500.0 mg, 36%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=553.2.
To a solution of N-[3-(5-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]-1,1-diphenylmethanimine (420.0 mg, 0.76 mmol) in DCM (5.0 mL) was added HCOOH (0.03 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (65/35, v/v) to afford 3-(5-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-amine (256.0 mg, 87%) as a white solid. LCMS (ESI, m/z): [M+H]+=389.2.
To a solution of 3-(5-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-amine (140.0 mg, 0.36 mmol) in DCM (5.0 mL) was added pyridine (112.0 mg, 1.41 mmol) and phenyl chloroformate (67.7 mg, 0.43 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. The resulting mixture was concentrated under reduced pressure. Then 2-(4-methylpiperazin-1-yl)ethanamine (206.4 mg, 1.44 mmol) and pyridine (5.0 ml) were added to the residue at 0° C. under N2. The resulting mixture was stirred at 60° C. for another 1 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (92/8, v/v) to afford 3-[3-(5-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (200.0 mg, 99%) as a white solid. LCMS (ESI, m/z): [M+H]+=558.3.
To a solution of 1-(3-(5-fluoro-2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)-3-(2-(4-methylpiperazin-1-yl)ethyl)urea (130.0 mg, 0.25 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 55% B in 10 min; 254 nm) to afford 3-[3-(5-fluoro-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (9.7 mg, 6%) as a white solid. LCMS (ESI, m/z): [M+H]+=428.3. 1H NMR (400 MHz, DMSO-d6): δ 13.37 (s, 1H), 9.64 (s, 1H), 8.43 (s, 1H), 8.02 (d, J=8.8 Hz, 1H), 7.42-7.39 (m, 1H), 7.28-7.13 (m, 3H), 3.82 (s, 3H), 3.30 (s, 3H), 3.05-2.83 (m, 4H), 2.79-2.59 (m, 8H).
To a solution of 3-(5-fluoro-2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (180.0 mg, 0.46 mmol) in DCM (5.0 mL) was added pyridine (144.0 mg, 1.82 mmol) and phenyl chloroformate (89.9 mg, 0.57 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. The resulting mixture was concentrated under reduced pressure. Then (2-aminoethyl)dimethylamine (162.1 mg, 1.84 mmol) and pyridine (5.0 mL) were added to the residue. The resulting mixture was stirred at 60° C. for another 1 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (92/8, v/v) to afford 3-[2-(dimethylamino)ethyl]-1-[3-(5-fluoro-2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (110.0 mg, 47%) as a white solid. LCMS (ESI, m/z): [M+H]+=503.3.
To a solution of 3-[2-(dimethylamino)ethyl]-1-[3-(5-fluoro-2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (110.0 mg, 0.22 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in CH3CN (3.0 mL) and NH3·H2O (3 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 27% B to 57% B in 7 min; 254 nm) to afford 3-[2-(dimethylamino)ethyl]-1-[3-(5-fluoro-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]urea (24.0 mg, 29%) as a white solid. LCMS (ESI, m/z): [M+H]+=373.2. 1H NMR (400 MHz, DMSO-d6): δ 11.68 (d, J=2.0 Hz, 1H), 9.21 (s, 1H), 8.27 (s, 1H), 8.09 (d, J=8.8 Hz, 1H), 8.03 (d, J=2.8 Hz, 1H), 7.91-7.88 (m, 1H), 7.72 (d, J=2.8 Hz, 1H), 7.18 (d, J=8.8 Hz, 1H), 3.94 (s, 3H), 3.33-3.28 (m, 2H), 2.41-2.38 (m, 2H), 2.20 (s, 6H).
To a solution of (6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)boronic acid (300.0 mg, 0.92 mmol) in dioxane/H2O (16.0/4.0 mL) was added 6-bromo-7-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole (328.2 mg, 0.92 mmol), K2CO3 (380.8 mg, 2.76 mmol) and Pd(dppf)Cl2 (67.2 mg, 0.09 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-7-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole (300.0 mg, 58%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=559.2.
To a solution of 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-7-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole (280.0 mg, 0.50 mmol) in dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (258.1 mg, 2.50 mmol), Cs2CO3 (489.4 mg, 1.50 mmol), Brettphos (53.8 mg, 0.10 mmol) and BrettPhos Pd G3 (45.4 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)-2-fluoro-N-(3-(7-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (110.0 mg, 35%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=626.3.
To a solution of (1S,2S)-2-fluoro-N-(3-(7-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (100.0 mg, 0.16 mmol) in DCM (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in ACN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 21% to 43% in 7 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-(7-methoxy-1H-indazol-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (10.8 mg, 18%) as a white solid. LCMS (ESI, m/z): [M+H]+=366.2. 1H NMR (300 MHz, DMSO-d6): δ 13.32-13.29 (m, 1H), 11.71 (s, 1H), 10.67 (s, 1H), 8.10-8.06 (m, 2H), 7.93 (d, J=8.4 Hz, 1H), 7.67 (s, 1H), 7.55 (d, J=8.1 Hz, 1H), 7.32 (d, J=8.4 Hz, 1H), 5.07-4.81 (m, 1H), 3.68 (s, 3H), 2.28-2.21 (m, 1H), 1.71-1.62 (m, 1H), 1.24-1.19 (m, 1H).
To a mixture of 6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl) ethoxy]methyl]indazole (800.0 mg, 1.97 mmol) and 6-chloro-3-iodo-1-[[2-(trimethylsilyl)ethoxy]methyl] pyrazolo[3,4-b]pyridine (810.5 mg, 1.97 mmol) in dioxane/H2O (10.0 mL/2.0 mL) was added K2CO3 (820.2 mg, 5.93 mmol) and Pd(dppf)Cl2 (144.7 mg, 0.19 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction was diluted with water at room temperature, extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/EtOAc (3/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-3-yl)-6-methoxy-1-[[2-(trimethylsilyl) ethoxy]methyl]indazole (520.0 mg, 47%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=560.2
To a mixture of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-3-yl)-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazole (260.0 mg, 0.46 mmol) and (1S,2S)-2-fluorocyclopropane-1-carboxamide (239.2 mg, 2.32 mmol) in t-BuOH (5.0 mL) was added Pd(OAc)2 (10.4 mg, 0.04 mmol), K2CO3 (192.4 mg, 1.39 mmol) and Xphos (25.5 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h under N2. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/EtOAc (3/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl] indazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (220.0 mg, 75%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=627.3.
To a solution of (1S,2S)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]indazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (200.0 mg, 0.31 mmol) in DCM (3.0 mL) was added TFA (3.0 mL). The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added ACN (1.0 mL) and NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 23% B to 33% B in 9 min; 254/220 nm) to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1H-indazol-5-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (12.2 mg, 10%) as a white solid. LCMS (ESI, m/z): [M+H]+=367.1. 1H NMR (300 MHz, DMSO-d6): δ 13.38 (s, 1H), 12.94 (s, 1H), 10.95 (s, 1H), 8.08-7.91 (m, 4H), 7.11 (s, 1H), 5.07-4.83 (m, 1H), 3.87 (s, 3H), 2.27-2.20 (m, 1H), 1.74-1.63 (m, 1H), 1.24-1.18 (m, 1H).
To a solution of 5-methoxy-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]imidazo[4,5-b]pyridine (500.0 mg, 1.23 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (446.1 mg, 1.23 mmol), K2CO3 (511.4 mg, 3.70 mmol) and Pd(dppf)Cl2 (180.5 mg, 0.25 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to afford 6-chloro-3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]imidazo[4,5-b]pyridin-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 43%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=560.2.
To a solution of 6-chloro-3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]imidazo[4,5-b]pyridin-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.50 mmol) in dioxane (4.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (154.8 mg, 1.49 mmol), BrettPhos (53.6 mg, 0.10 mmol), Cs2CO3 (488.5 mg, 1.49 mmol) and BrettPhos Pd G3 (45.3 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (39/61, v/v) to afford (1S,2S)-2-fluoro-N-[3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]imidazo[4,5-b]pyridin-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (300.0 mg, 76%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=627.3.
To a solution of (1S,2S)-2-fluoro-N-[3-(5-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]imidazo[4,5-b]pyridin-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (280.0 mg, 0.44 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in CH3CN (2.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure, then the was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 21% B to 29% B in 10 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-[5-methoxy-3H-imidazo[4,5-b]pyridin-6-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (8.1 mg, 5%) as a white solid. LCMS (ESI, m/z): [M+H]+=367.1. 1H NMR (400 MHz, DMSO-d6): δ 13.18-12.18 (m, 1H), 11.60 (s, 1H), 10.61 (s, 1H), 8.35-7.75 (m, 4H), 7.70-7.55 (m, 1H), 5.05-4.70 (m, 1H), 3.95 (s, 3H), 2.29-2.15 (m, 1H), 1.71-1.50 (m, 1H), 1.21-1.08 (m, 1H).
To a solution of 5-bromo-2-fluoro-4-methoxybenzaldehyde (500.0 mg, 2.14 mmol) in DMF (10.0 mL) was added (tert-butylsulfanyl)sodium (361.0 mg, 3.21 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 5-bromo-2-(tert-butylsulfanyl)-4-methoxybenzaldehyde (600.0 mg, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=303.0.
To a solution of 5-bromo-2-(tert-butylsulfanyl)-4-methoxybenzaldehyde (600.0 mg, 1.98 mmol) in EtOH/H2O (3.0/9.0 mL) was added NH2OH HCl (206.2 mg, 2.96 mmol) and NaOH (237.4 mg, 5.93 mmol) at 0° C. under N2. The resulting mixture was stirred at 60° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford (E)-N-[[5-bromo-2-(tert-butylsulfanyl)-4-methoxyphenyl]methylidene]hydroxylamine (510.0 mg, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=318.0.
To a solution of (E)-N-[[5-bromo-2-(tert-butylsulfanyl)-4-methoxyphenyl]methylidene]hydroxylamine (500.0 mg, 1.57 mmol) in toluene (5.0 mL) was added TsOH (54.1 mg, 0.31 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (65/35, v/v) to afford 5-bromo-6-methoxy-1,2-benzothiazole (150.0 mg, 39%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=243.9.
To a solution of 5-bromo-6-methoxy-1,2-benzothiazole (130.0 mg, 0.53 mmol) in dioxane/H2O (5.0/1.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (261.2 mg, 0.64 mmol), K2CO3 (220.8 mg, 1.60 mmol) and Pd(dppf)Cl2 (40.0 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1,2-benzothiazole (220.0 mg, 92%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=446.1.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1,2-benzothiazole (200.0 mg, 0.44 mmol) in 1,4-dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (230.0 mg, 2.24 mmol), Cs2CO3 (438.2 mg, 1.34 mmol), BrettPhos (48.1 mg, 0.09 mmol) and BrettPhos Pd G3 (40.6 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (65/35, v/v) to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1,2-benzothiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (83.0 mg, 36%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=513.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(6-methoxy-1,2-benzothiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (83.0 mg, 0.16 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in ACN (3.0 mL) was added NH3·H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 24% B to 54% B in 10 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1,2-benzothiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (4.0 mg, 6%) as a white solid. LCMS (ESI, m/z): [M+H]+=383.2. 1H NMR (400 MHz, DMSO-d6): δ 11.68 (d, J=2.0 Hz, 1H), 10.66 (s, 1H), 9.00 (d, J=0.8 Hz, 1H), 8.31 (s, 1H), 8.10 (d, J=8.8 Hz, 1H), 7.94-7.89 (m, 2H), 7.68 (d, J=2.0 Hz, 1H), 5.01-4.84 (m, 1H), 3.94 (s, 3H), 2.30-2.23 (m, 1H), 1.69-1.63 (m, 1H), 1.20-1.10 (m, 1H).
To a solution of 3-bromo-4-methoxyaniline (4.0 g, 19.80 mmol) in HOAc (30.0 mL) was added ammonium thiocyanate (7.6 g, 99.84 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 1 h. Then Br2 (1.2 mL) was added dropwise to the mixture at 0° C. The resulting mixture was stirred at 0° C. for another 2 h. After the reaction was completed, the reaction mixture was quenched with sat. NH4Cl (aq.) at 0° C. The pH value of the mixture was adjusted to 12 with NH3·H2O (aq.). The resulting mixture was extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (20/80, v/v) to afford 5-bromo-6-methoxy-1,3-benzothiazol-2-amine (3.2 g, 62%) as an off white solid. LCMS (ESI, m/z): [M+H]+=258.9.
To a solution of 5-bromo-6-methoxy-1,3-benzothiazol-2-amine (2.7 g, 10.42 mmol) in THE (10.0 mL) was added DMSO (90.0 mg, 1.15 mmol) and t-BuONO (1.8 g, 17.45 mmol) at room temperature. The resulting mixture was stirred at 30° C. for 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (88/12, v/v) to afford 5-bromo-6-methoxy-1,3-benzothiazole (2.3 g, 90%) as a white solid. LCMS (ESI, m/z): [M+H]+=243.9.
To a solution of 5-bromo-6-methoxy-1,3-benzothiazole (500.0 mg, 2.04 mmol) in dioxane (5.0 mL) was added bis(pinacolato)diboron (1.04 g, 4.09 mmol), KOAc (603.1 mg, 6.14 mmol) and Pd(dppf)Cl2 (149.8 mg, 0.20 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography with MeOH/H2O (1/1, v/v) to afford 6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-benzothiazole (300.0 mg, 50%) as a white solid. LCMS (ESI, m/z): [M+H]+=292.1.
To a solution of 6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3-benzothiazole (300.0 mg, 1.03 mmol) in dioxane/H2O (5.0/1.0 mL) was added 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (447.2 mg, 1.23 mmol), K2CO3 (599.5 mg, 4.33 mmol) and Pd(dppf)Cl2 (105.5 mg, 0.14 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (88/12, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1,3-benzothiazole (240.0 mg, 52%) as a white solid. LCMS (ESI, m/z): [M+H]+=446.1.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1,3-benzothiazole (200.0 mg, 0.44 mmol) in 1,4-dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (140.0 mg, 1.35 mmol), BrettPhos (48.1 mg, 0.09 mmol), Cs2CO3 (438.2 mg, 1.34 mmol) and BrettPhos Pd G3 (40.6 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (50/50, v/v) to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1,3-benzothiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (150.0 mg, 65%) as a white solid. LCMS (ESI, m/z): [M+H]+=513.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(6-methoxy-1,3-benzothiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (130.0 mg, 0.25 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 55% B in 10 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1,3-benzothiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (36.4 mg, 38%) as a white solid. LCMS (ESI, m/z): [M+H]+=383.2. 1H NMR (300 MHz, DMSO-d6): δ 11.66 (s, 1H), 10.64 (s, 1H), 9.22 (s, 1H), 8.14 (s, 1H), 8.01 (d, J=8.4 Hz, 1H), 7.93-7.87 (m, 2H), 7.66 (d, J=2.7 Hz, 1H), 5.06-4.79 (m, 1H), 3.90 (s, 3H), 2.26-2.20 (m, 1H), 1.73-1.63 (m, 1H), 1.29-1.09 (m, 1H).
To a solution of 4-bromo-3-methoxyaniline (2.0 g, 9.90 mmol) in Me2CO (50.0 mL) was added benzoyl isothiocyanate (1.6 g, 9.90 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 30 min. After the reaction was completed, the reaction mixture was cooled to room temperature and filtered. The solid was collected and dried to afford N-((4-bromo-3-methoxyphenyl)carbamothioyl)benzamide (3.1 g, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=365.0.
To a solution of N-((4-bromo-3-methoxyphenyl)carbamothioyl)benzamide (3.1 g, 8.49 mmol) in MeOH/H2O (30.0/30.0 mL) was added NaOH (0.4 g, 9.34 mmol) at room temperature. The resulting mixture was stirred at 65° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 1-(4-bromo-3-methoxyphenyl)thiourea (1.9 g, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=261.0.
To a solution of 1-(4-bromo-3-methoxyphenyl)thiourea (1.9 g, 7.28 mmol) in CHCl3 (30.0 mL) was added Br2 (1.4 g, 8.73 mmol) dropwise at 0° C. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the mixture was filtered. The solid was collected and dried to afford 6-bromo-5-methoxybenzo[d]thiazol-2-amine (1.8 g, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=258.9.
To a solution of 6-bromo-5-methoxybenzo[d]thiazol-2-amine (1.8 g, 6.95 mmol) in THE (30.0 mL) was added t-BuONO (1.1 g, 10.42 mmol) and DMSO (43.4 mg, 0.56 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 6-bromo-5-methoxybenzo[d]thiazole (470.0 mg, 27%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=243.9.
To a solution of 6-bromo-5-methoxybenzo[d]thiazole (200.0 mg, 0.82 mmol) in dioxane/H2O (10.0/2.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (334.9 mg, 0.82 mmol), K2CO3 (339.7 mg, 2.46 mmol) and Pd(dppf)Cl2 (60.0 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-methoxybenzo[d]thiazole (210.0 mg, 57%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=446.1.
To a solution of 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-methoxybenzo[d]thiazole (190.0 mg, 0.43 mmol) in dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (219.6 mg, 2.13 mmol), Cs2CO3 (416.4 mg, 1.28 mmol), Brettphos (45.7 mg, 0.09 mmol) and BrettPhos Pd G3 (38.6 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the reaction mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)-2-fluoro-N-(3-(5-methoxybenzo[d]thiazol-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (80.0 mg, 36%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=513.2.
To a solution of (1S,2S)-2-fluoro-N-(3-(5-methoxybenzo[d]thiazol-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (60.0 mg, 0.12 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 30% to 40% in 9 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-(5-methoxybenzo[d]thiazol-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (22.1 mg, 49%) as a white solid. LCMS (ESI, m/z): [M+H]+=383.1. 1H NMR (300 MHz, DMSO-d6): δ 11.67 (s, 1H), 10.66 (s, 1H), 9.33 (s, 1H), 8.28 (s, 1H), 8.10 (d, J=8.7 Hz, 1H), 7.92 (d, J=8.7 Hz, 1H), 7.76 (s, 1H), 7.69 (d, J=2.1 Hz, 1H), 5.07-4.80 (m, 1H), 3.94 (s, 3H), 2.29-2.20 (m, 1H), 1.73-1.62 (m, 1H), 1.22-1.15 (m, 1H).
A mixture of 2-bromo-4-fluoro-5-nitrophenol (3.0 g, 12.71 mmol), K2CO3 (3.5 g, 25.42 mmol) and MeI (3.6 g, 25.42 mmol) in acetone (20.0 mL) was stirred at 50° C. for 16 h. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure to afford 1-bromo-5-fluoro-2-methoxy-4-nitrobenzene (6.0 g, crude) as a yellow solid.
To a solution of 1-bromo-5-fluoro-2-methoxy-4-nitrobenzene (3.2 g, 12.79 mmol) in THF/H2O (10.0/10.0 mL) was added NaOH (767.8 mg, 19.19 mmol). The resulting mixture was stirred at 60° C. for 16 h. After the reaction was completed, the PH value of the mixture was adjusted to 6 with HCl (aq.). The resulting mixture was extracted with dichloromethane. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum to afford 5-bromo-4-methoxy-2-nitrophenol (2.5 g, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=247.9.
To a solution of 4-bromo-5-methoxy-2-nitrophenol (400.0 mg, 1.61 mmol) in MeOH (10.0 mL) was added Raney Ni (400.6 mg, 4.67 mmol). The mixture was stirred at room temperature for 2 h under H2. After the reaction was completed, the reaction mixture was filtered. The filtrate was concentrated under vacuum to afford 2-amino-5-bromo-4-methoxyphenol (400.0 mg, crude) as a brown solid. LCMS (ESI, m/z): [M+H]+=218.0.
A mixture of 2-amino-4-bromo-5-methoxyphenol (400.0 mg, 1.83 mmol) and triethyl orthoformate (399.6 mg, 2.69 mmol) in toluene (5.0 mL) was stirred at 100° C. for 16 h. The resulting mixture was concentrated under vacuum. The residue was diluted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (70/30, v/v) to afford 6-bromo-5-methoxybenzo[d]oxazole (300.0 mg, 72%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=228.0.
To a mixture of 6-bromo-5-methoxy-1,3-benzoxazole (200.0 mg, 0.87 mmol) and 6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-ylboronic acid (315.1 mg, 0.96 mmol) in dioxane/H2O (5.0/0.5 mL) was added Pd(dppf)Cl2 (71.6 mg, 0.09 mmol) and K2CO3 (363.6 mg, 2.63 mmol). The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by reverse phase flash column chromatography with H2O/ACN (50/50, v/v) to afford 6-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-methoxy-1,3-benzoxazole (180.0 mg, 47%) as a brown yellow oil. LCMS (ESI, m/z): [M+H]+=430.1.
To a mixture of 6-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-methoxy-1,3-benzoxazole (90.0 mg, 0.21 mmol) and (1S,2S)-2-fluorocyclopropane-1-carboxamide (107.9 mg, 1.05 mmol) in 1,4-dioxane (5.0 mL) was added BrettPhos Pd G3 (37.9 mg, 0.04 mmol), Cs2CO3 (204.6 mg, 0.63 mmol) and BrettPhos (44.9 mg, 0.08 mmol). The resulting mixture was stirred at 100° C. for 16 h under N2. The reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by reverse phase flash column chromatography with H2O/ACN (36/64, v/v) to afford (1S,2S)-2-fluoro-N-[3-(5-methoxy-1,3-benzoxazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (35.0 mg, 33%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=497.2.
A mixture of (1S,2S)-2-fluoro-N-[3-(5-methoxy-1,3-benzoxazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy] methyl]pyrrolo [2,3-b]pyridin-6-yl] cyclopropane-1-carboxamide (35.0 mg, 0.07 mmol) and TFA (2.0 mL) in DCM (2.0 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added ACN (2.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for another 4 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions: Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water, Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 5% B to 80% B in 7 min; 254/220 nm to afford (1S,2S)-2-fluoro-N-(3-(5-methoxybenzo[d]oxazol-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (3.9 mg, 15%) as a white solid. LCMS (ESI, m/z): [M+H]+=367.1. 1H NMR (300 MHz, DMSO-d6): δ 11.64 (s, 1H), 10.63 (s, 1H), 8.67 (s, 1H), 8.01 (d, J=8.7 Hz, 1H), 7.93-7.88 (m, 1H), 7.84 (s, 1H), 7.62 (d, J=2.4 Hz, 1H), 7.48 (s, 1H), 5.05-4.77 (m, 1H), 3.86 (s, 3H), 2.29-2.20 (m, 1H), 1.71-1.57 (m, 1H), 1.22-1.08 (m, 1H).
To a solution of 1-bromo-4-fluoro-2-methoxy-5-nitrobenzene (500.0 mg, 2.00 mmol) in H2O (20.0 mL) was added lithium hydroxide (239.4 mg, 9.99 mmol). The resulting mixture was stirred at 100° C. for 15 min. After the reaction was completed, the resulting mixture was diluted with water. The pH value of the mixture was adjusted to 5 with 1N HCl. The mixture was extracted with ethyl acetate. The combined organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum to afford 4-bromo-5-methoxy-2-nitrophenol (480.0 mg, crude) as a yellow solid.
To a solution of 4-bromo-5-methoxy-2-nitrophenol (400.0 mg, 1.61 mmol) in MeOH (10.0 mL) was added Raney Ni (400.6 mg, 4.67 mmol). The resulting mixture was stirred at room temperature for 2 h under H2. After the reaction was completed, the reaction mixture was filtered. The filtrate was concentrated under vacuum to afford 2-amino-4-bromo-5-methoxyphenol (400.0 mg, crude) as a brown yellow solid. LCMS (ESI, m/z): [M+H]+=218.0.
A mixture of 2-amino-4-bromo-5-methoxyphenol (400.0 mg, 1.83 mmol) and triethyl orthoformate (399.6 mg, 2.69 mmol) in toluene (5.0 mL) was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was diluted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (70/30, v/v) to afford 5-bromo-6-methoxybenzo[d]oxazole (300.0 mg, 72%) as a brown solid. LCMS (ESI, m/z): [M+H]+=228.0.
To a mixture of 5-bromo-6-methoxy-1,3-benzoxazole (300.0 mg, 1.31 mmol) and 6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-ylboronic acid (537.8 mg, 1.31 mmol) in 1,4-dioxane (5.0 mL) and H2O (0.5 mL) was added Pd(dppf)Cl2 (192.5 mg, 0.26 mmol) and K2CO3 (545.4 mg, 3.94 mmol). The mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (50/50, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1,3-benzoxazole (280.0 mg, 49%) as a brown yellow oil. LCMS (ESI, m/z): [M+H]+=430.1.
To a mixture of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1,3-benzoxazole (240.0 mg, 0.55 mmol) and (1S,2S)-2-fluorocyclopropane-1-carboxamide (575.4 mg, 5.58 mmol) in 1,4-dioxane (5.0 mL) was added BrettPhos (119.8 mg, 0.22 mmol), Pd2(dba)3 (102.2 mg, 0.11 mmol) and Cs2CO3 (545.5 mg, 1.67 mmol). The mixture was stirred at 100° C. for 16 h under N2. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by reverse phase flash chromatography with ACN/water (30/70, v/v) to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1,3-benzoxazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (77.0 mg, 27%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=497.2
A mixture of (1S,2S)-2-fluoro-N-[3-(6-methoxy-1,3-benzoxazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (67.0 mg, 0.14 mmol) and TFA (1.0 mL) in DCM (1.0 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added ACN (1.00 mL) and NH3·H2O (1.00 mL). The resulting mixture was stirred at room temperature for another 4 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions: Column: Xselect CSH OBD Column 30×150 mm 5 um; Mobile Phase A: Water, Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 5% B to 80% B in 7 min; 254/220 nm to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1,3-benzoxazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (14.7 mg, 29%) as a white solid. LCMS (ESI, m/z): [M+H]+=367.1 1H NMR (300 MHz, DMSO-d6): δ 11.59 (s, 1H), 10.62 (s, 1H), 8.62 (s, 1H), 7.94-7.89 (m, 2H), 7.80 (s, 1H), 7.56-7.54 (m, 2H), 5.01-4.80 (m, 1H), 3.86 (s, 3H), 2.28-2.17 (m, 1H), 1.86-1.62 (m, 1H), 1.18-0.83 (m, 1H).
To a solution of 3-bromo-6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in THF (10.0 mL) was added dropwise LDA (2.1 mL) at −70° C. under N2. The resulting mixture was stirred at −70° C. for 2 h. Then CH3I (588.6 mg, 4.15 mmol) was added dropwise to the mixture at −70° C. under N2. The resulting mixture was stirred at −70° C. for another 2 h. After the reaction was completed, the reaction was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford 3-bromo-6-chloro-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (320.0 mg, 61%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=375.0.
To a solution of 3-bromo-6-chloro-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (300.0 mg, 0.80 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added (2-methoxypyridin-3-yl)boronic acid (122.1 mg, 0.80 mmol), K2CO3 (331.0 mg, 2.40 mmol) and Pd(dppf)Cl2 (64.3 mg, 0.09 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford 6-chloro-3-(2-methoxypyridin-3-yl)-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (190.0 mg, 38%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=404.1.
To a solution of 6-chloro-3-(2-methoxypyridin-3-yl)-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (170.0 mg, 0.42 mmol) in dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (216.9 mg, 2.10 mmol), Cs2CO3 (411.9 mg, 1.26 mmol), Brettphos (45.2 mg, 0.08 mmol) and Brettphos Pd G3 (38.2 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (77/23, v/v) to afford (1S,2S)-2-fluoro-N-(3-(2-methoxypyridin-3-yl)-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (80.0 mg, 12%) as an off-white solid. LCMS (ESI, m/z): [M+H]+=471.2.
To a solution of (1S,2S)-2-fluoro-N-(3-(2-methoxypyridin-3-yl)-2-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (80.0 mg, 0.17 mmol) in DCM (1.5 mL) was added TFA (1.5 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in ACN (1.5 mL) was added NH3·H2O (1.5 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 35% B to 40% B in 13 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-(2-methoxypyridin-3-yl)-2-methyl-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (5.6 mg, 9%) as a white solid. LCMS (ESI, m/z): [M+H]+=341.2. 1H NMR (300 MHz, DMSO-d6): δ 11.45 (s, 1H), 10.56 (s, 1H), 8.17-8.15 (m, 1H), 7.81 (d, J=8.4 Hz, 1H), 7.71-7.68 (m, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.11-7.07 (m, 1H), 5.03-4.78 (m, 1H), 3.86 (s, 3H), 2.32 (s, 3H), 2.26-2.15 (m, 1H), 1.71-1.58 (m, 1H), 1.18-1.09 (m, 1H).
To a solution of 7-methoxy-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (500.0 mg, 0.62 mmol) in 1,4-dioxane/H2O (10.0 mL/2.0 mL) was added 6-chloro-3-iodo-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (211.1 mg, 0.52 mmol), K2CO3 (213.6 mg, 1.55 mmol) and Pd(dppf)Cl2 (37.7 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with ACN/H2O (9/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-3-yl)-4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (200.0 mg, 69%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=560.2.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-3-yl)-4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (150.0 mg, 0.27 mmol) in 1,4-dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (138.0 mg, 1.34 mmol), BrettPhos (28.7 mg, 0.05 mmol), Cs2CO3 (261.7 mg, 0.80 mmol) and BrettPhos Pd G3 (24.3 mg, 0.03 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/12, v/v) to afford (1S,2S)-2-fluoro-N-[3-(4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (79.0 mg, 47%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=627.3.
To a solution of (1S,2S)-2-fluoro-N-[3-(4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (110.0 mg, 0.18 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 11% B to 32% B in 7 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(4-methoxy-1H-1,3-benzodiazol-5-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (8.8 mg, 14%) as a white solid. LCMS (ESI, m/z): [M+H]+=367.2. 1H NMR (400 MHz, DMSO-d6): δ 13.36 (s, 1H), 12.61 (s, 1H), 10.96 (s, 1H), 8.28-8.23 (m, 1H), 8.15 (d, J=8.8 Hz, 1H), 7.99-7.96 (m, 1H), 7.50-7.44 (m, 1H), 7.27 (d, J=8.4 Hz, 1H), 5.04-4.85 (m, 1H), 4.29 (s, 3H), 2.33-2.23 (m, 1H), 1.73-1.62 (m, 1H), 1.23-1.15 (m, 1H).
To a solution of (1S,2R)-2-(hydroxymethyl)-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (34.0 mg, 0.07 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in CH3CN (2.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure, then the was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 19×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL/min; Gradient: 31% B to 40% B in 10 min; 254 nm) to afford (1S,2R)-2-(hydroxymethyl)-N-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (2.7 mg, 11%) as a white solid. LCMS (ESI, m/z): [M+H]+=338.2. 1H NMR (400 MHz, DMSO-d6): δ 11.50 (s, 1H), 10.44 (s, 1H), 7.96-7.87 (m, 2H), 7.54-7.51 (m, 2H), 7.28-7.24 (m, 1H), 7.10 (d, J=8.0 Hz, 1H), 7.03-6.99 (m, 1H), 4.45-4.42 (m, 1H), 3.80 (s, 3H), 3.67-3.61 (m, 1H), 3.55-3.49 (m, 1H), 2.12-2.07 (m, 1H), 1.45-1.39 (m, 1H), 0.99-0.85 (m, 2H).
To a mixture of 5-fluoro-2-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (300.0 mg, 1.75 mmol) and 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (634.8 mg, 1.75 mmol) in dioxane/H2O (10.0 mL/1.0 mL) was added Pd(dppf)Cl2 (128.4 mg, 0.17 mmol) and K2CO3 (727.7 mg, 5.26 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (20/1, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-fluoro-2-methoxypyridine (380.0 mg, 53%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=408.1
To a mixture of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-fluoro-2-methoxypyridine (280.0 mg, 0.68 mmol) and (1S,2S)-2-fluorocyclopropane-1-carboxamide (353.8 mg, 3.43 mmol) in BuOH (5.0 mL) was added Pd(OAc)2 (30.8 mg, 0.13 mmol), K2CO3 (284.5 mg, 2.05 mmol) and X-Phos (65.4 mg, 0.13 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) afford (1S,2S)-2-fluoro-N-[3-(5-fluoro-2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (170.0 mg, 52%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=475.2
To a solution of (1S,2S)-2-fluoro-N-[3-(5-fluoro-2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (170.0 mg, 0.35 mmol) in DCM (3.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added NH3·H2O (2.0 mL) and ACN (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 41% B to 50% B in 8 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(5-fluoro-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (37.0 mg, 30%) as a white solid. LCMS (ESI, m/z): [M+H]+=345.1. 1H NMR (300 MHz, DMSO-d6): δ 11.83 (s, 1H), 10.66 (s, 1H), 8.18 (d, J=8.7 Hz, 1H), 8.04 (d, J=3.0 Hz, 1H), 7.96-7.90 (m, 2H), 7.83 (d, J=2.7 Hz, 1H), 5.06-4.79 (m, 1H), 3.95 (s, 3H), 2.26-2.21 (m, 1H), 1.73-1.59 (m, 1H), 1.21-1.12 (m, 1H).
To a solution of 5-bromo-6-methoxy-1H-pyrrolo[2,3-b]pyridine (300.0 mg, 1.32 mmol) in CH2Cl2 (10.0 mL) was added Boc2O (285.8 mg, 1.32 mmol) and DMAP (160.0 mg, 1.32 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (92/8, v/v) to afford tert-butyl 5-bromo-6-methoxy-1H-pyrrolo[2,3-b]pyridine-1-carboxylate (350.0 mg, 81%) as a white solid. LCMS (ESI, m/z): [M+H]+=327.0.
To a solution of tert-butyl 5-bromo-6-methoxy-1H-pyrrolo[2,3-b]pyridine-1-carboxylate (320.0 mg, 0.98 mmol) in 1,4-dioxane/H2O (10.0/2.0 mL) was added (6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)boronic acid (319.5 mg, 0.98 mmol), K2CO3 (405.5 mg, 2.93 mmol) and Pd(dppf)Cl2 (71.5 mg, 0.10 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (94/6, v/v) to afford tert-butyl 6-chloro-6′-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H,1′H-[3,5′-bipyrrolo[2,3-b]pyridine]-1′-carboxylate (310.0 mg, 59%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=529.2.
To a solution of tert-butyl 6-chloro-6′-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H,1′H-[3,5′-bipyrrolo[2,3-b]pyridine]-1′-carboxylate (160.0 mg, 0.30 mmol) in 1,4-dioxane (6.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (155.9 mg, 1.51 mmol), K2CO3 (125.4 mg, 0.91 mmol), BrettPhos (32.5 mg, 0.06 mmol) and BrettPhos Pd G3 (27.4 mg, 0.03 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (78/22, v/v) to afford tert-butyl 6-((1S,2S)-2-fluorocyclopropane-1-carboxamido)-6′-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H,1′H-[3,5′-bipyrrolo[2,3-b]pyridine]-1′-carboxylate (180.0 mg, 99%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=596.3.
To a solution of tert-butyl 6-((1S,2S)-2-fluorocyclopropane-1-carboxamido)-6′-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H,1′H-[3,5′-bipyrrolo[2,3-b]pyridine]-1′-carboxylate (160.0 mg, 0.27 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xcelect CSH F-pheny OBD Column, 19×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: MeOH—Preparative; Flow rate: 25 mL/min; Gradient: 46% B to 61% B in 9 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(6′-methoxy-1H,1′H-[3,5′-bipyrrolo[2,3-b]pyridin]-6-yl)cyclopropane-1-carboxamide (18.8 mg, 19%) as a white solid. LCMS (ESI, m/z): [M+H]+=366.2. 1H NMR (400 MHz, DMSO-d6): δ 11.52-11.46 (m, 2H), 10.62 (s, 1H), 8.05-8.01 (m, 2H), 7.89 (d, J=8.4 Hz, 1H), 7.56 (d, J=2.4 Hz, 1H), 7.21-7.20 (m, 1H), 6.40-6.39 (m, 1H), 5.03-4.82 (m, 1H), 3.94 (s, 3H), 2.24-2.22 (m, 1H), 1.71-1.61 (m, 1H), 1.18-1.16 (m, 1H).
To a solution of 2-(2-cyclopropoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (320.0 mg, 1.23 mmol) in dioxane/H2O (16.0/4.0 mL) was added 6-chloro-3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (504.0 mg, 1.23 mmol), K2CO3 (510.0 mg, 3.69 mmol) and Pd(dppf)Cl2 (90.0 mg, 0.12 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 6-chloro-3-(2-cyclopropoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (300.0 mg, 58%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=416.1.
To a solution of 6-chloro-3-(2-cyclopropoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (260.0 mg, 0.63 mmol) in dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (322.2 mg, 3.13 mmol), Cs2CO3 (610.9 mg, 1.88 mmol), Brettphos (67.1 mg, 0.13 mmol) and BrettPhos Pd G3 (56.7 mg, 0.06 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford (1S,2S)—N-(3-(2-cyclopropoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (110.0 mg, 36%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=483.2.
To a solution of (1S,2S)—N-(3-(2-cyclopropoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (90.0 mg, 0.18 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XSelect CSH Prep C18 OBD Column 19×250 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: MeOH; Flow rate: 25 mL/min; Gradient: 57% to 69% in 12 min; 254 nm) to afford (1S,2S)—N-(3-(2-cyclopropoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (14.0 mg, 21%) as a white solid. LCMS (ESI, m/z): [M+H]+=353.2. 1H NMR (300 MHz, DMSO-d6): δ 13.48 (s, 1H), 10.98 (s, 1H), 8.06-7.96 (m, 2H), 7.63-7.60 (m, 1H), 7.50-7.46 (m, 2H), 7.12-7.07 (m, 1H), 5.08-4.84 (m, 1H), 3.93-3.89 (m, 1H), 2.29-2.25 (m, 1H), 1.72-1.63 (m, 1H), 1.23-1.16 (m, 1H), 0.83-0.66 (m, 4H).
To a solution of 2-bromo-4,5-difluorophenol (3.7 g, 17.94 mmol) in HOAc (20.0 mL) was added HNO3 (1.4 g, 21.5 mmol). The mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was quenched with water and extracted with ethyl acetate. The combined organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum to afford 6-bromo-3,4-difluoro-2-nitrophenol (3.0 g, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=253.9.
To a mixture of 6-bromo-3,4-difluoro-2-nitrophenol (2.0 g, 7.87 mmol) and K2CO3 (2.2 g, 15.75 mmol) in acetone (40.0 mL) was added methyl iodide (2.2 g, 15.75 mmol). The mixture was stirred at 70° C. for 16 h. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure to afford 1-bromo-4,5-difluoro-2-methoxy-3-nitrobenzene (2.0 g, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=267.9.
A mixture of 1-bromo-4,5-difluoro-2-methoxy-3-nitrobenzene (2.0 g, 7.46 mmol) in NH3/MeOH (16.0 mL, 7 mol/L) was stirred at 60° C. for 2 h. After the reaction was completed, the mixture was concentrated under reduced pressure to afford 4-bromo-6-fluoro-3-methoxy-2-nitroaniline (1.8 g, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=265.0.
To a mixture of 4-bromo-6-fluoro-3-methoxy-2-nitroaniline (1.0 g, 3.77 mmol) and NH4Cl (1.0 g, 18.86 mmol) in MeOH (50.0 mL) and H2O (5.0 mL) was added Fe (2.1 g, 37.73 mmol). The mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (50/50, v/v) to afford 4-bromo-6-fluoro-3-methoxybenzene-1,2-diamine (500.0 mg, 56%) as a brown oil. LCMS (ESI, m/z): [M+H]+=235.0.
A mixture of 4-bromo-6-fluoro-3-methoxybenzene-1,2-diamine (400.0 mg, 8.51 mmol) and triethyl orthoformate (5.0 mL) in acetic Acid (5.0 mL) was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum to afford 5-bromo-7-fluoro-4-methoxy-1H-1,3-benzodiazole (400.0 mg, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=245.0.
To a solution of 5-bromo-7-fluoro-4-methoxy-1H-1,3-benzodiazole (510.0 mg, 2.08 mmol) in THE (5.0 mL) was added sodium hydride (100.0 mg, 60%) at 0° C. The mixture was stirred at 0° C. for 15 min. Then SEM-Cl (381.8 mg, 2.29 mmol) was added to the mixture. The mixture was stirred at room temperature for another 2 h. After the reaction was completed, the reaction mixture was quenched with water and extracted with DCM. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (50/50, v/v) to afford 5-bromo-7-fluoro-4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (510.0 mg, 65%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=375.0.
To a solution of 5-bromo-7-fluoro-4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (400.0 mg, 1.06 mmol) in dioxane/H2O (10.0 mL/1.0 mL) was added 6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-ylboronic acid (382.9 mg, 1.17 mmol), K2CO3 (441.9 mg, 3.20 mmol) and Pd(dppf)Cl2 (77.9 mg, 0.11 mmol). The mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (94/6, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-7-fluoro-4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (330.0 mg, 54%) as a brown oil. LCMS (ESI, m/z): [M+H]+=577.2.
To a stirred mixture of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-7-fluoro-4-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,3-benzodiazole (250.0 mg, 0.43 mmol) and (1S,2S)-2-fluorocyclopropane-1-carboxamide (89.3 mg, 0.86 mmol) in 1,4-dioxane (5.0 mL) was added BrettPhos (92.9 mg, 0.17 mmol), BrettPhos Pd G3 (78.5 mg, 0.08 mmol) and Cs2CO3 (423.3 mg, 1.29 mmol). The resulting mixture was stirred at 100° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (94/6, v/v) to afford (1S,2S)-2-fluoro-N-(3-(7-fluoro-4-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-5-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (100.0 mg, 45%) as a brown solid. LCMS (ESI, m/z): [M+H]+=644.3.
A mixture of (1S,2S)-2-fluoro-N-(3-(7-fluoro-4-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-5-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (100.0 mg, 0.19 mmol) and TFA (2.0 mL) in DCM (2.0 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added ACN (2.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for another 4 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions: Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water, Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 5% B to 80% B in 7 min; 254/220 nm to afford (1S,2S)-2-fluoro-N-(3-(7-fluoro-4-methoxy-1H-benzo[d]imidazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (3.9 mg, 5%) as a white solid. LCMS (ESI, m/z): [M+H]+=384.1. 1H NMR (300 MHz, DMSO-d6): δ 13.09 (s, 1H), 11.67 (s, 1H), 10.64 (s, 1H), 8.25 (s, 1H), 8.09-8.02 (m, 1H), 7.90 (d, J=8.4 Hz, 1H), 7.69-7.46 (m, 1H), 7.16-7.13 (m, 1H), 5.05-4.77 (m, 1H), 4.09 (s, 1H), 3.58 (s, 2H), 2.29-2.18 (m, 1H), 1.71-1.57 (m, 1H), 1.22-1.08 (m, 1H).
To a solution of 5-bromo-6-methoxy-1H-1,2,3-benzotriazole (200.0 mg, 0.87 mmol) in THE (10.0 mmL) was added NaH (52.6 mg, 60%) at 0° C. The mixture was stirred at 0° C. for 15 min. Then SEM-Cl (160.8 mg, 0.96 mmol) was added to the mixture. The mixture was stirred at room temperature for another 2 h. After the reaction was completed, the reaction mixture was quenched with water and extracted with DCM. The organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (75/25, v/v) to afford 5-bromo-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,2,3-benzotriazole (220.0 mg, 70%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=358.1.
To a mixture of 5-bromo-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,2,3-benzotriazole (205.0 mg, 0.57 mmol) and 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (257.2 mg, 0.62 mmol) in dioxane (5.0 mL) and H2O (0.50 mL) was added Pd(dppf)Cl2 (83.7 mg, 0.11 mmol) and K2CO3 (237.2 mg, 1.71 mmol). The mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (50/50, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,2,3-benzotriazole (250.0 mg, 78%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=560.2.
To a mixture of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,2,3-benzotriazole (250.0 mg, 0.44 mmol) and (1S,2S)-2-fluorocyclopropane-1-carboxamide (230.0 mg, 2.23 mmol) in 1,4-dioxane (5.0 mL) was added BrettPhos (47.9 mg, 0.09 mmol), Pd2(dba)3 (163.4 mg, 0.18 mmol) and Cs2CO3 (436.1 mg, 1.33 mmol). The mixture was stirred at 100° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by reverse phase flash column chromatography with ACN/H2O (30/70, v/v) to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,2,3-benzotriazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (160.0 mg, 57%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=627.3.
A mixture of (1S,2S)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]-1,2,3-benzotriazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (150.0 mg, 0.24 mmol) and TFA (5.0 mL) in DCM (5.0 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added ACN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 4 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions: Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water, Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 5% B to 80% B in 7 min; 254/220 to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1H-1,2,3-benzotriazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (10.9 mg, 24%) as a white solid. LCMS (ESI, m/z): [M+H]+=367.1. 1H NMR (300 MHz, DMSO-d6): δ 15.33 (s, 1H), 11.63 (d, J=1.8 Hz, 1H), 10.63 (s, 1H), 7.99-7.88 (m, 3H), 7.61 (d, J=2.7 Hz, 1H), 7.35 (s, 1H), 5.05-4.77 (m, 1H), 3.89 (s, 3H), 2.26-2.20 (m, 1H), 1.74-1.54 (m, 1H), 1.22-1.05 (m, 1H).
To a solution of 3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (100.0 mg, 0.27 mmol) in DCM (5.0 mL) was added Pyridine (85.6 mg, 1.08 mmol) and phenyl carbonochloridate (50.8 mg, 0.32 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in pyridine (5.0 mL). Then (2-aminoethyl)dimethylamine (95.4 mg, 1.08 mmol) was added to the mixture at 0° C. under N2. The resulting mixture was stirred at 60° C. for another 1 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (92/8, v/v) to afford 1-(2-(dimethylamino)ethyl)-3-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)urea (100.0 mg, 77%) as a white solid. LCMS (ESI, m/z): [M+H]+=484.3.
To a solution of 1-(2-(dimethylamino)ethyl)-3-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)urea (100.0 mg, 0.21 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in ACN (3.0 mL) was added NH3—H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH F-Phenyl OBD column, 19×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: MeOH—Preparative; Flow rate: 25 mL/min; Gradient: 70% B to 90% B in 7 min; 254 nm) to afford 3-[2-(dimethylamino)ethyl]-1-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]urea (19.7 mg, 27%) as a white solid. LCMS (ESI, m/z): [M+H]+=354.2. 1H NMR (400 MHz, DMSO-d6): δ 11.41 (s, 1H), 9.16 (s, 1H), 8.37 (s, 1H), 7.90 (d, J=8.4 Hz, 1H), 7.51-7.49 (m, 1H), 7.45 (d, J=2.4 Hz, 1H), 7.28-7.23 (m, 1H), 7.11-7.07 (m, 2H), 7.03-6.99 (m, 1H), 3.80 (s, 3H), 3.32-3.28 (m, 2H), 2.41-2.38 (m, 2H), 2.20 (s, 6H).
To a solution of 6-chloro-3-iodo-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (300.0 mg, 0.73 mmol) in dioxane/H2O (5.0/1.0 mL) was added 5-fluoro-2-methoxyphenylboronic acid (149.3 mg, 0.88 mmol), K2CO3 (303.5 mg, 2.19 mmol) and Pd(dppf)Cl2 (53.5 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford 6-chloro-3-(5-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (220.0 mg, 74%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=408.1.
To a solution of 6-chloro-3-(5-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (160.0 mg, 0.38 mmol) in 1,4-dioxane (4.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (202.1 mg, 1.96 mmol), BrettPhos (42.1 mg, 0.08 mmol), Cs2CO3 (383.3 mg, 1.17 mmol) and BrettPhos Pd G3 (35.5 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (75/25, v/v) to afford (1S,2S)-2-fluoro-N-[3-(5-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (90.0 mg, 48%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=475.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(5-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (90.0 mg, 0.19 mmol) in CH3Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was re-dissolved in ACN (2.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 34% B to 44% B in 8 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(5-fluoro-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (12.1 mg, 18%) as a white solid. LCMS (ESI, m/z): [M+H]+=345.2. 1H NMR (400 MHz, DMSO-d6): δ 13.58 (s, 1H), 10.97 (s, 1H), 8.15 (d, J=8.8 Hz, 1H), 8.00 (d, J=8.8 Hz, 1H), 7.45 (d, J=2.4 Hz, 1H), 7.30-7.21 (m, 2H), 5.10-4.83 (m, 1H), 3.83 (s, 3H), 2.37-2.23 (m, 1H), 1.78-1.67 (m, 1H), 1.26-1.13 (m, 1H).
To a solution of 6-chloro-3-iodo-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (500.0 mg, 1.22 mmol) in 1,4-dioxane/H2O (25.0 mL/5.0 mL) was added 4-fluoro-2-methoxyphenylboronic acid (207.4 mg, 1.22 mmol), K2CO3 (337.3 mg, 2.44 mmol) and Pd(dppf)Cl2 (89.3 mg, 0.12 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 6-chloro-3-(4-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (347.0 mg, 70%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=408.1.
To a solution of 6-chloro-3-(4-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (347.0 mg, 0.85 mmol) in 1,4-dioxane (20.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (438.5 mg, 4.26 mmol), BrettPhos (91.3 mg, 0.17 mmol), Cs2CO3 (831.4 mg, 2.55 mmol) and BrettPhos Pd G3 (77.1 mg, 0.09 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(4-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (56.5 mg, 14%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=475.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(4-fluoro-2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (56.6 mg, 0.12 mmol) in DCM (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O. The pH value of the mixture was adjusted to 7 with aq·NaHCO3. The mixture was extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 36% B to 54% B in 8 min; 254/220 nm) to afford (1S,2S)-2-fluoro-N-[3-(4-fluoro-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (6.8 mg, 16%) as a white solid. LCMS (ESI, m/z): [M+H]+=345.3. 1H NMR (400 MHz, DMSO-d6): δ 13.50 (s, 1H), 10.96 (s, 1H), 8.10 (d, J=8.8 Hz, 1H), 7.98 (d, J=8.8 Hz, 1H), 7.65-7.61 (m, 1H), 7.12-7.08 (m, 1H), 6.92-6.87 (m, 1H), 5.04-4.86 (m, 1H), 3.85 (s, 3H), 2.35-2.25 (m, 1H), 1.70-1.64 (m, 1H), 1.22-1.16 (m, 1H).
To a solution of 6-chloro-3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (400.0 mg, 0.98 mmol) in 1,4-dioxane/H2O (8.0/2.0 mL) was added (3-fluoro-2-methoxyphenyl)boronic acid (165.9 mg, 0.98 mmol), K2CO3 (404.8 mg, 2.93 mmol) and Pd(dppf)Cl2 (71.4 mg, 0.10 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford 6-chloro-3-(3-fluoro-2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (370.0 mg, 88%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=408.1.
To a solution of 6-chloro-3-(3-fluoro-2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (310.0 mg, 0.76 mmol) in 1,4-dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (391.7 mg, 3.80 mmol), K2CO3 (315.1 mg, 2.28 mmol), Brettphos (81.6 mg, 0.15 mmol) and Brettphos Pd G3 (68.9 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (83/17, v/v) to afford (1S,2S)-2-fluoro-N-(3-(3-fluoro-2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)cyclopropane-1-carboxamide (106.0 mg, 28%) as a brown oil. LCMS (ESI, m/z): [M+H]+=475.2.
To a solution of (1S,2S)-2-fluoro-N-(3-(3-fluoro-2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)cyclopropane-1-carboxamide (86.0 mg, 0.18 mmol) in DCM (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 34% B to 51% B in 7 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-(3-fluoro-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)cyclopropane-1-carboxamide (12.2 mg, 19%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=345.2. 1H NMR (300 MHz, DMSO-d6): δ 13.67 (s, 1H), 11.02 (s, 1H), 8.21 (d, J=9.0 Hz, 1H), 8.03 (d, J=9.0 Hz, 1H), 7.52 (d, J=7.8 Hz, 1H), 7.42-7.35 (m, 1H), 7.28-7.21 (m, 1H), 5.09-4.83 (m, 1H), 3.70 (d, J=1.2 Hz, 3H), 2.47-2.32 (m, 1H), 1.74-1.61 (m, 1H), 1.26-1.16 (m, 1H).
To a solution of 6-chloro-3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (400.0 mg, 0.97 mmol) in 1,4-dioxane/H2O (10.0 mL/2.0 mL) was added (2-hydroxyphenyl)boronic acid (134.7 mg, 0.97 mmol), K2CO3 (404.8 mg, 2.93 mmol) and Pd(dppf)Cl2 (71.4 mg, 0.10 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (75/25, v/v) to afford 2-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenol (270.0 mg, 74%) as a light green solid. LCMS (ESI, m/z): [M+H]+=376.1.
To a solution of 2-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)phenol (220.0 mg, 0.59 mmol) in DMF (5.0 mL) was added K2CO3 (242.6 mg, 1.76 mmol) and iodomethane-d3 (127.3 mg, 0.88 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (99/1, v/v) to afford 6-chloro-3-(2-(methoxy-d3)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (210.0 mg, 91%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=393.2.
To a solution of 6-chloro-3-(2-(methoxy-d3)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (120.0 mg, 0.31 mmol) in 1,4-dioxane (6.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (157.4 mg, 1.53 mmol), Cs2CO3 (298.5 mg, 0.92 mmol), BrettPhos (32.8 mg, 0.06 mmol) and BrettPhos Pd G3 (27.7 mg, 0.03 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (75/25, v/v) to afford (1S,2S)-2-fluoro-N-(3-(2-(methoxy-d3)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)cyclopropane-1-carboxamide (75.0 mg, 53%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=460.2.
To a solution of (1S,2S)-2-fluoro-N-(3-(2-(methoxy-d3)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)cyclopropane-1-carboxamide (75.0 mg, 0.16 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in ACN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Shield RP18 OBD Column, 19×250 mm, 10 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 25 mL/min; Gradient: 39% B to 49% B in 10 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-(2-(methoxy-d3)phenyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)cyclopropane-1-carboxamide (10.9 mg, 20%) as a white solid. LCMS (ESI, m/z): [M+H]+=330.1. 1H NMR (300 MHz, DMSO-d6): δ 13.47 (s, 1H), 10.97 (s, 1H), 8.12 (d, J=9.0 Hz, 1H), 7.99 (d, J=9.0 Hz, 1H), 7.64-7.61 (m, 1H), 7.47-7.41 (m, 1H), 7.19 (d, J=7.5 Hz, 1H), 7.10-7.05 (m, 1H), 5.08-4.82 (m, 1H), 2.30-2.25 (m, 1H), 1.73-1.62 (m, 1H), 1.23-1.16 (m, 1H).
To a solution of 6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-ylboronic acid (350.0 mg, 1.07 mmol) in dioxane/H2O (5.0/1.0 mL) was added 5-bromo-4,6-dimethoxypyrimidine (187.7 mg, 0.85 mmol), K2CO3 (444.2 mg, 3.20 mmol) and Pd(dppf)Cl2 (78.4 mg, 0.10 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4,6-dimethoxypyrimidine (180.0 mg, 40%) as a white solid. LCMS (ESI, m/z): [M+H]+=421.1.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4,6-dimethoxypyrimidine (160.0 mg, 0.38 mmol) in 1,4-dioxane (4.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (195.9 mg, 1.90 mmol), Cs2CO3 (371.5 mg, 1.14 mmol), BrettPhos (40.8 mg, 0.07 mmol) and BrettPhos Pd G3 (34.4 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (70/30, v/v) to afford (1S,2S)—N-[3-(4,6-dimethoxypyrimidin-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (160.0 mg, 86%) as a white solid. LCMS (ESI, m/z): [M+H]+=488.2.
To a solution of (1S,2S)—N-[3-(4,6-dimethoxypyrimidin-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (280.0 mg, 0.44 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in ACN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 31% B to 40% B in 9 min; 254 nm) to afford (1S,2S)—N-[3-(4,6-dimethoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (29.5 mg, 25%) as a white solid. LCMS (ESI, m/z): [M+H]+=358.1. 1H NMR (400 MHz, DMSO-d6): δ 11.66 (s, 1H), 10.62 (s, 1H), 8.47 (s, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.71 (d, J=8.4 Hz, 1H), 7.49 (d, J=1.6 Hz, 1H), 5.01-4.82 (m, 1H), 3.91 (s, 6H), 2.27-2.19 (m, 1H), 1.69-1.61 (m, 1H), 1.17-1.14 (m, 1H).
To a mixture of 6-chloro-3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.77 mmol) and ethylurea (206.0 mg, 0.23 mmol) in dioxane (5.0 mL) was added Brettphos Pd G3 (69.9 mg, 0.07 mmol), Cs2CO3 (753.8 mg, 2.31 mmol) and BrettPhos (84.0 mg, 0.14 mmol) at room temperature under N2. The resulting mixture was stirred at 60° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 3-ethyl-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl) ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (300.0 mg, 88%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=441.2
To a solution of 3-ethyl-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl] pyrrolo[2,3-b]pyridin-6-yl]urea (170.0 mg, 0.38 mmol) in DCM (2.0 mL) was added TFA (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added ACN (1.0 mL) and NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 41% B to 50% B in 9 min; 254 nm) to afford 3-ethyl-1-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]urea (43.9 mg, 36%) as a white solid. LCMS (ESI, m/z): [M+H]+=311.2. 1H NMR (300 MHz, DMSO-d6): δ 11.51 (s, 1H), 9.15 (s, 1H), 8.59-8.56 (m, 1H), 7.91 (d, J=8.4 Hz, 1H), 7.52-7.46 (m, 2H), 7.29-7.23 (m, 1H), 7.10 (d, J=7.5 Hz, 1H), 7.04-6.99 (m, 2H), 3.81 (s, 3H), 3.30-3.21 (m, 2H), 1.19-1.14 (m, 3H).
To a solution of 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (100.0 mg, 0.27 mmol) in CH2Cl2 (2.0 mL) was added Pyridine (85.6 mg, 1.08 mmol) and phenyl chloroformate (50.8 mg, 0.33 mmol) at 0° C. The mixture was stirred at room temperature for 16 h. The mixture was concentrated under vacuum. To the residue was added pyridine (2.0 mL) and 3-aminopropanenitrile (56.9 mg, 0.81 mmol). The resulting mixture was stirred at 60° C. for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (20/80, v/v) to afford 3-(2-cyanoethyl)-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (70.0 mg, 55%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=466.2.
To a solution of 3-(2-cyanoethyl)-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (60.0 mg, 0.13 mmol) in CH2Cl2 (1.0 mL) was added TFA (1.0 mL). The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. The residue was dissolved in ACN (1.0 mL) and NH3·H2O (1.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 32% B to 47% B in 7 min; 254 nm) to afford 3-(2-cyanoethyl)-1-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]urea (24.5 mg, 56%) as a white solid. LCMS (ESI, m/z): [M+H]+=336.1. 1H NMR (300 MHz, DMSO-d6): δ 11.48 (s, 1H), 9.33 (s, 1H), 8.70 (s, 1H), 7.93 (d, J=8.4 Hz, 1H), 7.53-7.48 (m, 2H), 7.29-7.24 (m, 1H), 7.12-7.08 (m, 2H), 7.04-6.99 (m, 1H), 3.81 (s, 3H), 3.51-3.46 (m, 2H), 2.80-2.73 (m, 2H).
To a solution of trimethylsulfoxonium iodide (1.2 g, 5.61 mmol) in DMSO (10.0 mL) was added NaH (224.6 mg, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. Then 5,6-dihydropyran-2-one (500.0 mg, 5.10 mmol) was added dropwise to the mixture at 0° C. The resulting mixture was stirred at room temperature for another 16 h under N2. After the reaction was completed, the resulting mixture was quenched with water and extracted with Et2O. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford cis-3-oxa-bicyclo[4.1.0]heptan-2-one (80.0 mg, crude) as a yellow oil.
To a mixture of 3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (70.0 mg, 0.19 mmol) and cis-3-oxa-bicyclo[4.1.0]heptan-2-one (63.7 mg, crude) in THE (10.0 mL) was added AlMe3 (0.47 mL, 2 mol/L) at 0° C. under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (30/70, v/v) to afford cis-2-(2-hydroxyethyl)-N-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (40.0 mg, 44%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=482.2.
To a solution of cis-2-(2-hydroxyethyl)-N-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (35.0 mg, 0.07 mmol) in CH2Cl2 (1.0 mL) was added TFA (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in ACN (1.0 mL) and NH3·H2O (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with petroleum MeOH/H2O (70/30, v/v) to afford cis-2-(2-hydroxyethyl)-N-(3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropanecarboxamide (10.3 mg, 40%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=352.2. 1H NMR (300 MHz, DMSO-d6): δ 11.53 (d, J=1.8 Hz, 1H), 10.47 (s, 1H), 7.98-7.88 (m, 2H), 7.56-7.52 (m, 2H), 7.30-7.24 (m, 1H), 7.13-7.05 (m, 1H), 7.03-7.00 (m, 1H), 4.44-4.40 (m, 1H), 3.82 (s, 3H), 3.43-3.37 (m, 2H), 2.07-2.01 (m, 1H), 1.72-1.63 (m, 2H), 1.33-1.25 (m, 1H), 0.99-0.85 (m, 2H).
To a solution of 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (300.0 mg, 0.82 mmol) in DMF (10.0 mL) was added cis-2-cyanocyclopropane-1-carboxylic acid (90.9 mg, 0.81 mmol), HATU (463.2 mg, 1.21 mmol) and DIEA (157.3 mg, 1.28 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford cis-2-cyano-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (300.0 mg, 80%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=463.2.
To a solution of cis-2-cyano-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (150.0 mg, 0.43 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 55% B to 75% B in 10 min; 254 nm) to afford cis-2-cyano-N-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (3.7 mg, 3%) as a white solid. LCMS (ESI, m/z): [M+H]+=333.1. 1H NMR (300 MHz, DMSO-d6): δ 11.66 (s, 1H), 10.93 (s, 1H), 8.01 (d, J=8.4 Hz, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.60 (d, J=2.1 Hz, 1H), 7.55-7.52 (m, 1H), 7.31-7.25 (m, 1H), 7.12 (d, J=7.5 Hz, 1H), 7.05-7.00 (m, 1H), 3.82 (s, 3H), 2.77-2.71 (m, 1H), 2.16-2.10 (m, 1H), 1.63-1.54 (m, 1H), 1.45-1.38 (m, 1H).
To a solution of trans-2-(hydroxymethyl)-N-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (300.0 mg, 0.64 mmol) in THF (10.0 mL) was added PPh3 (185.0 mg, 0.70 mmol) and CBr4 (319.2 mg, 0.96 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (83/17, v/v) to afford trans-2-(bromomethyl)-N-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (250.0 mg, 40%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=530.1.
To a solution of trans-2-(bromomethyl)-N-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (200.0 mg, 0.39 mmol) in DMSO (10.0 mL) was added KCN (37.7 mg, 0.58 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 3 h. After the reaction was completed, the resulting mixture was quenched with aq. NaHCO3 and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (72/28, v/v) to afford trans-2-(cyanomethyl)-N-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (150.0 mg, 81%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=477.2.
To a solution of trans-2-(cyanomethyl)-N-(3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (130.0 mg, 0.27 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 42% B to 52% B in 8 min; 254 nm) to afford trans-2-(cyanomethyl)-N-(3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (37.2 mg, 39%) as a white solid. LCMS (ESI, m/z): [M+H]+=347.2. 1H NMR (300 MHz, DMSO-d6): δ 11.58 (s, 1H), 10.70 (s, 1H), 7.98 (d, J=8.7 Hz, 1H), 7.88 (d, J=8.7 Hz, 1H), 7.58-7.52 (m, 2H), 7.30-7.25 (m, 1H), 7.11 (d, J=8.1 Hz, 1H), 7.05-7.00 (m, 1H), 3.82 (s, 3H), 2.73 (d, J=6.6 Hz, 2H), 2.10-2.05 (m, 1H), 1.59-1.52 (m, 1H), 1.14-1.08 (m, 1H), 0.95-0.83 (m, 1H).
To a solution of 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy] methyl]pyrrolo[2,3-b] pyridin-6-amine (600.0 mg, 1.62 mmol) in DMF (10.0 mL) was added DIEA (1049.2 mg, 8.12 mmol), trans-2-(methoxycarbonyl) cyclopropane-1-carboxylic acid (280.8 mg, 1.95 mmol) and HATU (926.0 mg, 2.44 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (50/50, v/v) to afford trans-methyl 2-[[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]cyclopropane-1-carboxylate (660.0 mg, 82%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=496.2.
To a solution of trans-methyl 2-[[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl) ethoxy] methyl] pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]cyclopropane-1-carboxylate (640.0 mg, 1.29 mmol) in THF/CH3OH (8.0/2.0 mL) was added NaBH4 (488.5 mg, 12.91 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the reaction mixture was quenched with CH3OH and concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (50/50, v/v) to afford trans-2-(hydroxymethyl)-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl) ethoxy]methyl]pyrrolo[2,3-b] pyridin-6-yl]cyclopropane-1-carboxamide (500.0 mg, 82%) as a white solid. LCMS (ESI, m/z): [M+H]+=468.2.
To a solution of trans-2-(hydroxymethyl)-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy] methyl]pyrrolo [2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (130.0 mg, 0.28 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 29% B to 59% B in 7 min; 254 nm) to afford trans-2-(hydroxymethyl)-N-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b] pyridin-6-yl]cyclopropane-1-carboxamide (20.3 mg, 21%) as a white solid. LCMS (ESI, m/z): [M+H]+=338.2. 1H NMR (300 MHz, DMSO-d6): δ 11.54 (s, 1H), 10.46 (s, 1H), 7.96 (d, J=8.7 Hz, 1H), 7.87 (d, J=8.7 Hz, 1H), 7.55-7.52 (m, 2H), 7.29-7.23 (m, 1H), 7.10 (d, J=7.5 Hz, 1H), 7.04-6.99 (m, 1H), 4.66-4.62 (m, 1H), 3.81 (s, 3H), 3.49-3.42 (m, 1H), 3.31-3.27 (m, 1H), 1.97-1.93 (m, 1H), 1.49-1.43 (m, 1H), 1.01-0.95 (m, 1H), 0.80-0.75 (m, 1H).
Step 1: Synthesis of tert-butyl N-[2-(4-ethylpiperazin-1-yl)ethyl]carbamate
To a solution of tert-butyl N-[2-(piperazin-1-yl)ethyl]carbamate (2.0 g, 8.72 mmol) in CH3CN (20.0 mL) was added K2CO3 (2.4 g, 17.44 mmol) and ethyl iodide (2.0 g, 13.08 mmol). The mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (88/12, v/v) to afford tert-butyl N-[2-(4-ethylpiperazin-1-yl)ethyl]carbamate (400.0 mg, 17%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=258.2.
The solution of tert-butyl N-[2-(4-ethylpiperazin-1-yl)ethyl]carbamate (400.0 mg, 1.55 mmol) in HCl/dioxane (3.0 mL, 4 mol/L) was stirred at room temperature for 2 h. After the reaction was completed, the mixture was filtered. The solid was washed with Et2O and dried to afford 2-(4-ethylpiperazin-1-yl)ethanamine hydrochloride (200.0 mg, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=158.2.
To a solution of 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (150.0 mg, 0.41 mmol) in CH2Cl2 (3.0 mL) was added pyridine (128.4 mg, 1.62 mmol) and phenyl chloroformate (76.3 mg, 0.49 mmol) at 0° C. The mixture was stirred at room temperature for 16 h. The mixture was concentrated under vacuum. To the above residue was added pyridine (3.0 mL) and 2-(4-ethylpiperazin-1-yl)ethanamine hydrochloride (236.7 mg, crude). The resulting mixture was stirred at 60° C. for another 5 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/95, v/v) to afford 1-[2-(4-ethylpiperazin-1-yl)ethyl]-3-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (200.0 mg, 89%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=553.3
To a solution of 1-[2-(4-ethylpiperazin-1-yl)ethyl]-3-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (180.0 mg, 0.33 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL). The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. To the above residue was added ACN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 37% B to 47% B in 8 min; 254 nm) to afford 1-[2-(4-ethylpiperazin-1-yl)ethyl]-3-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]urea (24.6 mg, 17%) as a white solid. LCMS (ESI, m/z): [M+H]+=423.2. 1H NMR (400 MHz, DMSO-d6): δ 11.42 (s, 1H), 9.18 (s, 1H), 8.30 (s, 1H), 7.90 (d, J=8.4 Hz, 1H), 7.51-7.49 (m, 1H), 7.45 (d, J=2.0 Hz, 1H), 7.28-7.23 (m, 1H), 7.10 (d, J=8.4 Hz, 2H), 7.03-6.99 (m, 1H), 3.81 (s, 3H), 2.47-2.44 (m, 6H), 2.36-2.26 (m, 6H), 0.99-0.96 (m, 3H).
To a solution of tert-butyl N-[2-(piperidin-4-yl)ethyl]carbamate (1.0 g, 4.38 mmol) in THE (50.0 mL) was added 3-oxetanone (0.4 g, 5.26 mmol) and NaBH(OAc)3 (1.4 g, 6.57 mmol at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) to afford tert-butyl N-[2-[1-(oxetan-3-yl)piperidin-4-yl]ethyl]carbamate (900.0 mg, 72%) as a brown solid. LCMS (ESI, m/z): [M+H]+=285.0.
To a solution of tert-butyl N-[2-[1-(oxetan-3-yl)piperidin-4-yl]ethyl]carbamate (800.0 mg, 2.81 mmol) in DCM (6.0 mL) was added TFA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the mixture was basified to pH=8 with saturated NaHCO3 (aq.). The mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 2-[1-(oxetan-3-yl)piperidin-4-yl]ethanamine (500.0 mg, crude) as a brown oil. LCMS (ESI, m/z): [M+H]+=185.2.
To a solution of 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (130.0 mg, 0.35 mmol) in DCM (5.0 mL) was added pyridine (112.7 mg, 1.41 mmol) and phenyl chloroformate (66.1 mg, 0.42 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added pyridine (6.0 mL) and 2-[1-(oxetan-3-yl)piperidin-4-yl]ethanamine (129.7 mg, 0.70 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 16 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (5/1, v/v) to afford 3-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1-[2-[4-(oxetan-3-yl)piperazin-1-yl]ethyl]urea (100.0 mg, 49%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=581.3.
To a solution of 3-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1-[2-[4-(oxetan-3-yl)piperazin-1-yl]ethyl]urea (90.0 mg, 0.16 mmol) in CH2Cl2 (4.0 mL) was added TFA (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (4.0 mL) and NH3·H2O (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 55% B in 9 min; 254 nm) to afford 3-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-1-[2-[1-(oxetan-3-yl)piperidin-4-yl]ethyl]urea (2.2 mg, 3%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=450.3. 1H NMR (400 MHz, DMSO-d6): δ 11.49 (s, 1H), 9.13 (s, 1H), 8.55 (s, 1H), 7.89 (d, J=8.4 Hz, 1H), 7.53 (s, 1H), 7.45 (d, J=2.4 Hz, 1H), 7.26-7.22 (m, 1H), 7.11-7.08 (m, 1H), 7.02-6.95 (m, 2H), 4.52-4.49 (m, 2H), 4.42-4.39 (m, 2H), 3.80 (s, 3H), 3.30-3.25 (m, 2H), 2.68-2.64 (m, 2H), 1.79-1.69 (m, 4H), 1.49-1.44 (m, 2H), 1.37-1.28 (m, 1H), 1.24-1.17 (m, 2H).
To a solution of 5-bromo-4-methoxypyridin-2-amine (1.0 g, 4.93 mmol) in MeOH (10.0 mL) was added DMF-DMA (704.2 mg, 5.91 mmol) at room temperature. The resulting mixture was stirred at 75° C. for 3 h. After the reaction was completed, the resulting mixture concentrated under reduced pressure to afford (E)-N′-(5-bromo-4-methoxypyridin-2-yl)-N,N-dimethylformimidamide (870.0 mg, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=258.0.
To a solution of (E)-N′-(5-bromo-4-methoxypyridin-2-yl)-N,N-dimethylformimidamide (550.0 mg, 2.13 mmol) in CH3OH/Pyridine (5.0/0.5 mL) was added (aminooxy)sulfonic acid (361.5 mg, 3.20 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was quenched with aq. NaHCO3 and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (1/1, v/v) to afford 6-bromo-7-methoxy-[1,2,4]triazolo[1,5-a]pyridine (100.0 mg, 20%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=228.0.
To a solution of 6-bromo-7-methoxy-[1,2,4]triazolo[1,5-a]pyridine (150.0 mg, 0.66 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (268.9 mg, 0.66 mmol), K2CO3 (272.7 mg, 1.97 mmol) and Pd(dppf)Cl2 (48.1 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/90, v/v) to afford 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-7-methoxy-[1,2,4]triazolo[1,5-a]pyridine (100.0 mg, 35%) as a light brown solid. LCMS (ESI, m/z): [M+H]+=430.1.
To a solution of 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-7-methoxy-[1,2,4]triazolo[1,5-a]pyridine (80.0 mg, 0.19 mmol) in 1,4-dioxane (4.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (95.9 mg, 0.93 mmol), Cs2CO3 (181.9 mg, 0.56 mmol), BrettPhos (20.0 mg, 0.04 mmol) and BrettPhos Pd G3 (16.9 mg, 0.02 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (30/70, v/v) to afford (1S,2S)-2-fluoro-N-(3-(7-methoxy-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (70.0 mg, 76%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=497.2.
To a solution of (1S,2S)-2-fluoro-N-(3-(7-methoxy-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (70.0 mg, 0.14 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 23% B to 33% B in 8 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-(7-methoxy-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (14.3 mg, 27%) as a white solid. LCMS (ESI, m/z): [M+H]+=367.2. 1H NMR (300 MHz, DMSO-d6): δ 11.77 (s, 1H), 10.68 (s, 1H), 8.90 (s, 1H), 8.35 (s, 1H), 8.02-7.91 (m, 2H), 7.68 (d, J=2.4 Hz, 1H), 7.36 (s, 1H), 5.08-4.72 (m, 1H), 3.96 (s, 3H), 2.30-2.13 (m, 1H), 1.78-1.55 (m, 1H), 1.23-1.04 (m, 1H).
To a solution of 6-chloro-3-iodo-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (400.0 mg, 0.98 mmol) in dioxane/H2O (10.0 mL/2.0 mL) was added 2-methoxy-4-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (243.2 mg, 0.98 mmol), K2CO3 (404.8 mg, 2.93 mmol) and Pd(dppf)Cl2 (71.4 mg, 0.10 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-3-yl)-2-methoxy-4-methylpyridine (100.0 mg, 25%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=405.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-3-yl)-2-methoxy-4-methylpyridine (160.0 mg, 0.40 mmol) in 1,4-dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (203.7 mg, 1.98 mmol), BrettPhos (42.4 mg, 0.08 mmol), Cs2CO3 (386.2 mg, 1.19 mmol) and BrettPhos Pd G3 (35.8 mg, 0.04 mmol) at room temperature under N2. The reaction mixture was stirred with microwave at 120° C. for 1.5 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(2-methoxy-4-methylpyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (57.0 mg, 31%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=472.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(2-methoxy-4-methylpyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (57.0 mg, 0.12 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 33% B to 38% B in 8 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(2-methoxy-4-methylpyridin-3-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (10.9 mg, 26%) as a white solid. LCMS (ESI, m/z): [M+H]+=342.1. 1H NMR (400 MHz, DMSO-d6): δ 13.53 (s, 1H), 10.99 (s, 1H), 8.13 (s, 1H), 7.98-7.90 (m, 2H), 7.02 (s, 1H), 5.03-4.87 (m, 1H), 3.79 (s, 3H), 2.27-2.20 (m, 4H), 1.70-1.64 (m, 1H), 1.30-1.15 (m, 1H).
To a solution of 3-bromo-6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in 1,4-dioxane/H2O (25.0 mL/5.0 mL) was added 2-methoxy-4-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (413.2 mg, 1.66 mmol), K2CO3 (382.1 mg, 2.76 mmol) and Pd(PPh3)4 (159.7 mg, 0.14 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-methoxy-4-methylpyridine (160.0 mg, 28%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=404.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-methoxy-4-methylpyridine (140.0 mg, 0.35 mmol) in 1,4-dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (178.6 mg, 1.73 mmol), BrettPhos (37.2 mg, 0.07 mmol), Cs2CO3 (338.7 mg, 1.04 mmol) and BrettPhos Pd G3 (31.4 mg, 0.04 mmol) at room temperature under N2. The reaction mixture was stirred with microwave at 120° C. for 1.5 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford (1S,2S)-2-fluoro-N-[3-(2-methoxy-4-methylpyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (80.0 mg, 49%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=471.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(2-methoxy-4-methylpyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (80.0 mg, 0.17 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 34% B to 44% B in 8 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(2-methoxy-4-methylpyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (6.9 mg, 11%) as a white solid. LCMS (ESI, m/z): [M+H]+=341.0. 1H NMR (400 MHz, DMSO-d6): δ 11.59 (s, 1H), 10.62 (s, 1H), 8.02 (d, J=4.8 Hz, 1H), 7.84 (d, J=8.8 Hz, 1H), 7.55 (d, J=8.4 Hz, 1H), 7.38 (d, J=2.4 Hz, 1H), 6.97 (d, J=5.2 Hz, 1H), 5.02-4.81 (m, 1H), 3.75 (s, 3H), 2.23-2.18 (m, 4H), 1.68-1.61 (m, 1H), 1.19-1.14 (m, 1H).
To a solution of 4-bromopyridin-3-ol (5.0 g, 28.74 mmol) in sulfuric acid (25.0 mL) was added nitric acid (5.4 g, 86.21 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (9/1, v/v) to afford 4-bromo-2-nitropyridin-3-ol (2.3 g, 19%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=218.9.
To a solution of 4-bromo-2-nitropyridin-3-ol (2.3 g, 10.50 mmol) in DMF (115.0 mL) was added K2CO3 (2.9 g, 21.01 mmol) at room temperature. The resulting mixture was stirred at room temperature for 10 min. Then CH3I (3.0 g, 21.11 mmol) was added to the mixture at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (9/1, v/v) to afford 4-bromo-3-methoxy-2-nitropyridine (1.8 g, 73%) as a white solid LCMS (ESI, m/z): [M+H]+=232.9.
To a solution of 4-bromo-3-methoxy-2-nitropyridine (1.8 g, 7.73 mmol) in AcOH (10.0 mL)/EtOH (10.0 mL)/H2O (5.0 mL) was added Fe (2.2 g, 38.68 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography petroleum ether/ethyl acetate (1/1, v/v) to afford 4-bromo-3-methoxypyridin-2-amine (1.3 g, 83%) as a white solid. LCMS (ESI, m/z): [M+H]+=203.1.
To a solution of 4-bromo-3-methoxypyridin-2-amine (650.0 mg, 3.20 mmol) in sat NaHCO3 (25.0 mL)/DCM (25.0 mL) was added chloroacetaldehyde (1.6 g, 8.00 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the reaction mixture was diluted with water and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 7-bromo-8-methoxyimidazo[1,2-a]pyridine (450.0 mg, 62%) as a white solid. LCMS (ESI, m/z): [M+H]+=227.0.
To a solution of 7-bromo-8-methoxyimidazo[1,2-a]pyridine (500.0 mg, 2.20 mmol) in dioxane/H2O (20.0 mL/4.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (900.2 mg, 2.20 mmol), K2CO3 (913.0 mg, 6.61 mmol), Pd(dppf)Cl2 (161.1 mg, 0.22 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 6-chloro-3-[8-methoxyimidazo[1,2-a]pyridin-7-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (140.0 mg, 15%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=429.1.
To a solution of 6-chloro-3-[8-methoxyimidazo[1,2-a]pyridin-7-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (120.0 mg, 0.28 mmol) in 1,4-dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (144.2 mg, 1.40 mmol), BrettPhos (30.0 mg, 0.06 mmol), Cs2CO3 (273.4 mg, 0.84 mmol) and BrettPhos Pd G3 (25.4 mg, 0.03 mmol) at room temperature under N2. The resulting mixture was stirred with microwave at 120° C. for 90 min. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/2, v/v) to afford (1S,2S)-2-fluoro-N-(3-[8-methoxyimidazo[1,2-a]pyridin-7-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (70.0 mg, 50%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=496.2.
To a solution of (1S,2S)-2-fluoro-N-(3-[8-methoxyimidazo[1,2-a]pyridin-7-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (70.0 mg, 0.14 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 35% B in 8 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-[8-methoxyimidazo[1,2-a]pyridin-7-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (11.7 mg, 23%) as a white solid. LCMS (ESI, m/z): [M+H]+=366.2. 1H NMR (400 MHz, DMSO-d6): δ 11.78 (s, 1H), 10.67 (s, 1H), 8.35 (d, J=6.4 Hz, 1H), 8.14 (d, J=8.4 Hz, 1H), 7.98-7.92 (m, 2H), 7.74 (s, 1H), 7.56 (s, 1H), 7.15 (d, J=6.4 Hz, 1H), 5.02-4.85 (m, 1H), 4.12 (s, 3H), 2.25-2.20 (m, 1H), 1.69-1.64 (m, 1H), 1.22-1.10 (m, 1H).
To a solution of 4-bromopyridin-3-ol (10.0 g, 57.47 mmol) in H2SO4 (30.0 mL) was added HNO3 (10.0 mL) at 0° C. under N2. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 4-bromo-2-nitropyridin-3-ol (6.3 g, 35%) as an off-white solid. LCMS (ESI, m/z): [M+H]+=218.9.
To a solution of 4-bromo-2-nitropyridin-3-ol (6.3 g, 28.77 mmol) in DMF (20.0 mL) was added K2CO3 (7.9 g, 56.94 mmol) and CH3I (4.0 g, 28.77 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (93/7, v/v) to afford 4-bromo-3-methoxy-2-nitropyridine (3.2 g, 48%) as a white solid. LCMS (ESI, m/z): [M+H]+=232.9.
To a solution of 4-bromo-3-methoxy-2-nitropyridine (3.2 g, 13.73 mmol) in EtOH/H2O (25.0/5.0 mL) was added Fe (3.8 g, 68.66 mmol) and NH4Cl (3.6 g, 68.66 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with DCM/MeOH (95/5, v/v) to afford 4-bromo-3-methoxypyridin-2-amine (2.6 g, 93%) as a purple solid. LCMS (ESI, m/z): [M+H]+=203.0.
To a solution of 4-bromo-3-methoxypyridin-2-amine (2.5 g, 12.31 mmol) in CH3OH (40.0 mL) was added DMF-DMA (1.8 g, 14.94 mmol) at room temperature under N2. The resulting mixture was stirred at 70° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (45/55, v/v) to afford (E)-N′-(4-bromo-3-methoxypyridin-2-yl)-N,N-dimethylmethanimidamide (2.4 g, 75%) as a white solid. LCMS (ESI, m/z): [M+H]+=258.0.
To a solution of (E)-N′-(4-bromo-3-methoxypyridin-2-yl)-N,N-dimethylmethanimidamide (2.4 g, 9.30 mmol) in CH3OH (20.0 mL) was added NH2OH HCl (2.4 g, 34.51 mmol) at room temperature under N2. The resulting mixture was stirred at 70° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (77/23, v/v) to afford (E)-N′-(4-bromo-3-methoxypyridin-2-yl)-N-hydroxymethanimidamide (2.1 g, 92%) as a white solid. LCMS (ESI, m/z): [M+H]+=246.0.
To a solution of (E)-N′-(4-bromo-3-methoxypyridin-2-yl)-N-hydroxymethanimidamide (1.0 g, 4.06 mmol) in THF (5.0 mL) was added TFAA (938.9 mg, 4.47 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 7-bromo-8-methoxy-[1,2,4]triazolo[1,5-a]pyridine (1.2 g, crude) as a brown solid. LCMS (ESI, m/z): [M+H]+=228.0.
To a solution of 7-bromo-8-methoxy-[1,2,4]triazolo[1,5-a]pyridine (1.2 g, 5.26 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (2.4 g, 5.87 mmol), K2CO3 (2.1 g, 15.19 mmol) and Pd(dppf)Cl2 (420.0 mg, 0.54 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford 6-chloro-3-[8-methoxy-[1,2,4]triazolo[1,5-a]pyridin-7-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (1.2 g, 52%) as a white solid. LCMS (ESI, m/z): [M+H]+=430.1.
To a solution of 6-chloro-3-[8-methoxy-[1,2,4]triazolo[1,5-a]pyridin-7-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.68 mmol) in 1,4-dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (360.0 mg, 3.49 mmol), Cs2CO3 (681.9 mg, 2.01 mmol), Brettphos (75.0 mg, 0.14 mmol) and Brettphos Pd G3 (63.2 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (68/32, v/v) to afford (1S,2S)-2-fluoro-N-(3-[8-methoxy-[1,2,4]triazolo[1,5-a]pyridin-7-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (122.0 mg, 36%) as an off-white solid. LCMS (ESI, m/z): [M+H]+=497.2.
To a solution of (1S,2S)-2-fluoro-N-(3-[8-methoxy-[1,2,4]triazolo[1,5-a]pyridin-7-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (60.0 mg, 0.12 mmol) in DCM (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in ACN (3.0 mL) and NH3·H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 11% B to 41% B in 9 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-[8-methoxy-[1,2,4]triazolo[1,5-a]pyridin-7-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (14.4 mg, 32%) as a white solid. LCMS (ESI, m/z): [M+H]+=367.0. 1H NMR (400 MHz, DMSO-d6): δ 11.93 (s, 1H), 10.71 (s, 1H), 8.72 (d, J=7.2 Hz, 1H), 8.48 (s, 1H), 8.16 (d, J=8.8 Hz, 1H), 7.97 (d, J=8.4 Hz, 1H), 7.86 (s, 1H), 7.45 (d, J=7.2 Hz, 1H), 5.04-4.83 (m, 1H), 4.20 (s, 3H), 2.26-2.23 (m, 1H), 1.71-1.64 (m, 1H), 1.26-1.10 (m, 1H).
To a solution of 6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-ylboronic acid (300.0 mg, 0.92 mmol) in 1,4-dioxane/H2O (15.0 mL/3.0 mL) was added 3-bromo-5-fluoro-2,4-dimethoxypyridine (216.8 mg, 0.92 mmol), K2CO3 (253.9 mg, 1.84 mmol) and Pd(dppf)Cl2 (67.2 mg, 0.09 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-fluoro-2,4-dimethoxypyridine (277.9 mg, 69%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=438.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-fluoro-2,4-dimethoxypyridine (213.0 mg, 0.49 mmol) in 1,4-dioxane (16.0 mL) was added diphenylmethanimine (440.7 mg, 2.43 mmol), BrettPhos (52.2 mg, 0.10 mmol), Cs2CO3 (475.4 mg, 1.46 mmol) and BrettPhos Pd G3 (44.1 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to afford N-[3-(5-fluoro-2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1,1-diphenylmethanimine (146.9 mg, 51%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=583.2.
To a solution of N-[3-(5-fluoro-2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1,1-diphenylmethanimine (123.5 mg, 0.21 mmol) in CH2Cl2 (6.0 mL) was added FA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, The pH value of the mixture was adjusted to 8 with aq·NaHCO3. The resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford 3-(5-fluoro-2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (46.3 mg, 52%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=419.2.
To a solution of 3-(5-fluoro-2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (209.0 mg, 0.50 mmol) in DCM (9.0 mL) was added pyridine (158.0 mg, 2.00 mmol) and phenyl chloroformate (93.8 mg, 0.60 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. To the above mixture was added 2-(4-methylpiperazin-1-yl)ethanamine (214.6 mg, 1.50 mmol) and pyridine (9.0 mL) at room temperature. The resulting mixture was stirred at 60° C. for 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with methylene chloride/methanol (90/10, v/v) to afford 3-[3-(5-fluoro-2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (99.1 mg, 33%) as a light brown oil. LCMS (ESI, m/z): [M+H]+=588.3.
To a solution of 3-[3-(5-fluoro-2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (99.1 mg, 0.169 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 55% B in 7 min; 254 nm) to afford 3-[3-(5-fluoro-2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (10.6 mg, 13%) as a white solid. LCMS (ESI, m/z): [M+H]+=458.3. 1H NMR (400 MHz, DMSO-d6): δ 11.55 (s, 11H), 9.19 (s, 11H), 8.27 (s, 11H), 8.10 (d, J=2.8 Hz, 11H), 7.62 (d, J=8.8 Hz, 11H), 7.35 (s, 1H), 7.08 (d, J=8.8 Hz, 1H), 3.79-3.78 (m, 6H), 2.54-2.44 (m, 6H), 2.32 (s, 4H), 2.15 (s, 3H).
To a solution of 6-chloro-3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (270.0 mg, 0.64 mmol) in toluene (12.0 mL) was added diphenylmethanimine (350.4 mg, 1.93 mmol), XantPhos (74.6 mg, 0.13 mmol), t-BuOK (216.9 mg, 1.93 mmol) and Pd2(dba)3 (118.0 mg, 0.13 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (135.5 mg, 53%) as a light brown oil. LCMS (ESI, m/z): [M+H]+=400.2.
To a solution of 3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (180.0 mg, 0.45 mmol) in DCM (8.0 mL) was added Pyridine (142.5 mg, 1.80 mmol) and phenyl chloroformate (84.6 mg, 0.54 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added 2-(4-methylpiperazin-1-yl)ethanamine (193.6 mg, 1.35 mmol) and pyridine (8.0 mL) at room temperature. The resulting mixture was stirred at 60° C. for another 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with methylene chloride/methanol (9/1, v/v) to afford 3-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (209.2 mg, 82%) as a light brown oil. LCMS (ESI, m/z): [M+H]+=569.3.
To a solution of 3-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (209.2 mg, 0.37 mmol) in CH2Cl2 (6.0 mL) was added TFA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (6.0 mL) was added NH3·H2O (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 3 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10.0 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 47% B in 7 min; 254 nm) to afford 3-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (14.3 mg, 8%) as a white solid. LCMS (ESI, m/z): [M+H]+=439.2. 1H NMR (400 MHz, DMSO-d6): δ 11.28 (s, 1H), 9.13 (s, 1H), 8.37 (s, 1H), 7.46 (d, J=8.4 Hz, 1H), 7.28-7.24 (m, 1H), 7.16 (d, J=2.0 Hz, 1H), 6.99 (d, J=8.4 Hz, 1H), 6.73 (d, J=8.4 Hz, 2H), 3.68 (s, 6H), 2.50-2.44 (m, 6H), 2.32 (s, 4H), 2.15 (s, 3H).
To a solution of 6-chloro-3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (280.0 mg, 0.67 mmol) in dioxane (7.5 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (344.5 mg, 3.34 mmol), K2CO3 (277.1 mg, 2.0 mmol), XPhos (63.7 mg, 0.13 mmol) and Pd(OAc)2 (15.0 mg, 0.07 mmol) under N2. The mixture was stirred at 100° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/EtOAc (2/1, v/v) to afford (1S,2S)—N-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (129.0 mg, 39%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=486.2.
To a solution of (1S,2S)—N-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (109.0 mg, 0.22 mmol) in CH2Cl2 (6.0 mL) was added TFA (3.0 mL) at room temperature. The reaction mixture was stirred at room temperature for 4 h. The mixture was concentrated under vacuum. The residue was dissolved in CH3CN (5.0 mL) and NH3·H2O (5.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions: Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 34% B to 64% B in 7 min; 254 nm, RT1: 6.5 min to afford (1S,2S)—N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (24.9 mg, 30%) as a white solid. LCMS (ESI, m/z): [M+H]+=356.0. 1H NMR (300 MHz, DMSO-d6): δ 11.43 (s, 1H), 10.58 (s, 1H), 7.82 (d, J=8.4 Hz, 1H), 7.55 (d, J=8.7 Hz, 1H), 7.31-7.25 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 5.04-4.80 (m, 1H), 3.70 (s, 6H), 2.25-2.20 (m, 1H), 1.70-1.60 (m, 1H), 1.18-1.11 (m, 1H).
To a solution of 1-fluoro-3-iodo-2,4-dimethoxybenzene (1.0 g, 3.55 mmol) in 1,4-dioxane (18.0 mL) and H2O (3.6 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (1.5 g, 3.55 mmol), K2CO3 (1.0 g, 0.01 mmol) and Pd(dppf)Cl2 (0.3 g, 0.36 mmol) under N2. The mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/EtOAc (10/1, v/v) to afford 6-chloro-3-(3-fluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (1.0 g, 64%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=437.1.
To a solution of 6-chloro-3-(3-fluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.69 mmol) in t-BuOH (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (353.9 mg, 3.43 mmol), K2CO3 (284.7 mg, 2.06 mmol), XPhos (65.5 mg, 0.14 mmol) and Pd(OAc)2 (15.4 mg, 0.07 mmol) under N2. The mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/EtOAc (2/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(3-fluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (279.0 mg, 55%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=504.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(3-fluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (279.0 mg, 0.56 mmol) in CH2Cl2 (14.0 mL) was added TFA (14.0 mL) at room temperature. The reaction mixture was stirred at room temperature for 4 h. The mixture was concentrated under vacuum. The residue was dissolved in CH3CN (15.0 mL) and NH3·H2O (15.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions: Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 41% B to 51% B in 9 min; 254/220 nm; to afford (1S,2S)-2-fluoro-N-[3-(3-fluoro-2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (78.0 mg, 37%) as a white solid. LCMS (ESI, m/z): [M+H]+=374.0. 1H NMR (300 MHz, DMSO-d6): δ 11.60 (d, J=1.8 Hz, 1H), 10.62 (s, 1H), 7.84 (d, J=8.4 Hz, 1H), 7.65 (d, J=8.4 Hz, 1H), 7.40 (d, J=2.4 Hz, 1H), 7.24-7.17 (m, 1H), 6.85-6.80 (m, 1H), 5.06-4.78 (m, 1H), 3.68 (s, 3H), 3.53 (s, 3H), 2.25-2.20 (m, 1H), 1.70-1.59 (m, 1H), 1.18-1.11 (m, 1H)
To a solution of 1,5-difluoro-2,4-dimethoxybenzene (500.0 mg, 2.87 mmol) in HOAc (10.0 mL) was added NIS (1.3 g, 5.74 mmol) at room temperature. The mixture was stirred at 50° C. for 16 h. After the reaction was completed, the mixture was concentrated under vacuum. The residue was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/EtOAc (5/1, v/v) to afford 1,5-difluoro-3-iodo-2,4-dimethoxybenzene (386.0 mg, 44%) as a white solid.
To a solution of 1,5-difluoro-3-iodo-2,4-dimethoxybenzene (500.0 mg, 1.67 mmol) in 1,4-dioxane (10.0 mL) and H2O (2.0 mL) were added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (681.2 mg, 1.67 mmol), Pd(PPh3)4 (385.1 mg, 0.33 mmol) and K2CO3 (460.6 mg, 3.33 mmol) at room temperature under N2. The mixture was stirred at 100° C. for 12 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/EtOAc (10/1, v/v) to afford 6-chloro-3-(3,5-difluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (321.0 mg, 42%) as a white solid. LCMS (ESI, m/z): [M+H]+=455.1.
To a solution of 6-chloro-3-(3,5-difluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (260.0 mg, 0.57 mmol) in t-BuOH (5.5 mL) were added (1S,2S)-2-fluorocyclopropane-1-carboxamide (294.6 mg, 2.86 mmol), K2CO3 (236.9 mg, 1.71 mmol), Pd(OAc)2 (12.8 mg, 0.06 mmol) and XPhos (54.5 mg, 0.11 mmol) at room temperature under N2. The mixture was stirred at 100° C. for 12 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/EtOAc (2/1, v/v) to afford (1S,2S)—N-[3-(3,5-difluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (70.0 mg, 23%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=522.2.
To a solution of (1S,2S)—N-[3-(3,5-difluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (60.0 mg, 0.12 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The reaction mixture was stirred at room temperature for 4 h. The mixture was concentrated under vacuum. The residue was dissolved in CH3CN (4.0 mL) and NH3·H2O (4.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions: Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 42% B to 52% B in 9 min to afford (1S,2S)—N-[3-(3,5-difluoro-2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (14.9 mg, 32%) as a white solid. LCMS (ESI, m/z): [M+H]+=392.0. 1H NMR (300 MHz, DMSO-d6): δ 11.77 (s, 1H), 10.66 (s, 1H), 7.88 (d, J=8.7 Hz, 1H), 7.76 (d, J=8.4 Hz, 1H), 7.52 (d, J=2.4 Hz, 1H), 7.46-7.38 (m, 1H), 5.07-4.79 (m, 1H), 3.50 (s, 6H), 2.29-2.20 (m, 1H), 1.72-1.59 (m, 1H), 1.21-1.10 (m, 1H).
To a solution of 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (150.0 mg, 0.41 mmol) in CH2Cl2 (3.0 mL) was added pyridine (128.4 mg, 1.62 mmol) and phenyl chloroformate (76.3 mg, 0.49 mmol) at 0° C. The mixture was stirred at room temperature for 16 h. The mixture was concentrated under vacuum. To the above residue was added pyridine (3.0 mL) and tert-butyl 4-(2-aminoethyl)piperazine-1-carboxylate (279.3 mg, 1.22 mmol). The resulting mixture was stirred at 60° C. for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/95, v/v) to afford tert-butyl 4-[2-([[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (190.0 mg, 74%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=625.3.
To a solution of tert-butyl 4-[2-([[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (160.0 mg, 0.26 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL). The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. To the above residue was added ACN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 55% B in 7 min; 254 nm) to afford 3-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(piperazin-1-yl)ethyl]urea (48.0 mg, 47%) as a white solid. LCMS (ESI, m/z): [M+H]+=395.2. 1H NMR (400 MHz, DMSO-d6): δ 11.43 (s, 1H), 9.19 (s, 1H), 8.32 (s, 1H), 7.90 (d, J=8.8 Hz, 1H), 7.52-7.45 (m, 2H), 7.28-7.23 (m, 1H), 7.11-6.99 (m, 3H), 3.81 (s, 3H), 2.51-2.36 (m, 12H).
To a solution of 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (130.0 mg, 0.35 mmol) in DCM (10.0 mL) was added pyridine (111.3 mg, 1.41 mmol) and phenyl chloroformate (66.1 mg, 0.42 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the above mixture was added 1-cyclopropylmethanamine (75.1 mg, 1.06 mmol) and pyridine (10.0 mL) at room temperature. The resulting mixture was stirred at 70° C. for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (50/50, v/v) to afford 3-(cyclopropylmethyl)-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (160.0 mg, 97%) as a light brown oil. LCMS (ESI, m/z): [M+H]+=467.2.
To a solution of 3-(cyclopropylmethyl)-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (165.0 mg, 0.35 mmol) in CH2Cl2 (6.0 mL) was added TFA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (6.0 mL) and NH3·H2O (6.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 45% B to 53% B in 9 min; 254 nm) to afford 3-(cyclopropylmethyl)-1-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]urea (13.5 mg, 11%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=337.3. 1H NMR (400 MHz, DMSO-d6): δ 11.49 (s, 1H), 9.14 (s, 1H), 8.56 (s, 1H), 7.90 (d, J=8.8 Hz, 1H), 7.52-7.46 (m, 2H), 7.28-7.24 (m, 1H), 7.10 (d, J=8.0 Hz, 1H), 7.05-6.99 (m, 2H), 3.81 (s, 3H), 3.15-3.12 (m, 2H), 1.06-0.99 (m, 1H), 0.50-0.45 (m, 2H), 0.24-0.19 (m, 2H).
To a solution of 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (130.0 mg, 0.35 mmol) in DCM (10.0 mL) was added pyridine (111.3 mg, 1.41 mmol) and phenyl chloroformate (66.1 mg, 0.42 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. To the above mixture was added aminocyclopropane (60.3 mg, 1.06 mmol) and pyridine (10.0 mL) at room temperature. The resulting mixture was stirred at 70° C. for another 3 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (50/50, v/v) to afford 3-cyclopropyl-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (120.0 mg, 75%) as a light brown oil. LCMS (ESI, m/z): [M+H]+=453.2.
To a solution of 3-cyclopropyl-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (120.0 mg, 0.27 mmol) in CH2Cl2 (6.0 mL) was added TFA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (6.0 mL) and NH3·H2O (6.0 mL). The resulting mixture was stirred at room temperature for another 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 41% B to 51% B in 8 min; 254 nm) to afford 3-cyclopropyl-1-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]urea (35.8 mg, 41%) as a white solid. LCMS (ESI, m/z): [M+H]+=323.3. 1H NMR (400 MHz, DMSO-d6): δ 11.53 (s, 1H), 9.13 (s, 1H), 8.73 (s, 1H), 7.90 (d, J=8.8 Hz, 1H), 7.51-7.46 (m, 2H), 7.28-7.23 (m, 1H), 7.10 (d, J=7.6 Hz, 1H), 7.02-6.98 (m, 2H), 3.80 (s, 3H), 2.68-2.65 (m, 1H), 0.72-0.69 (m, 2H), 0.53-0.50 (m, 2H).
To a solution of tert-butyl N-[2-(2-hydroxyethoxy)ethyl]carbamate (2.0 g, 9.74 mmol) in CH2Cl2 (15.0 mL) were added p-toluenesulfonyl chloride (3.7 g, 19.49 mmol), TEA (4.9 g, 48.72 mmol) and DMAP (119.0 mg, 0.97 mmol). The mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (45/55, v/v) to afford tert-butyl N-(2-[2-[(4-methylbenzenesulfonyl)oxy]ethoxy]ethyl)carbamate (2.3 g, 65%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=360.1.
To a solution of tert-butyl N-(2-[2-[(4-methylbenzenesulfonyl)oxy]ethoxy]ethyl)carbamate (2.2 g, 6.12 mmol) in THF/H2O (20.0 mL/3.0 mL) was added dimethylamine hydrochloride (2.0 g, 24.48 mmol) and NaOH (0.9 g, 24.48 mmol). The mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (95/5, v/v) to afford tert-butyl N-[2-[2-(dimethylamino)ethoxy]ethyl]carbamate (1.2 g, 84%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=233.2.
A solution of tert-butyl N-[2-[2-(dimethylamino)ethoxy]ethyl]carbamate (1.2 g, 5.16 mmol) in HCl/1,4-dioxane (10.0 mL, 4 mol/L) was stirred at room temperature for 2 h. The mixture was concentrated under vacuum to afford 2-(2-aminoethoxy)-N,N-dimethylethan-1-amine hydrochloride (700.0 mg, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=133.1.
To a solution of 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (150.0 mg, 0.41 mmol) in CH2Cl2 (3.0 mL) was added pyridine (128.4 mg, 1.62 mmol) and phenyl chloroformate (76.3 mg, 0.49 mmol) at 0° C. The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. To the above mixture was added pyridine (3.0 mL) and 2-(2-aminoethoxy)-N,N-dimethylethan-1-amine hydrochloride (176.8 mg, crude). The resulting mixture was stirred at 60° C. for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (96/4, v/v) to afford 3-[2-[2-(dimethylamino)ethoxy]ethyl]-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (180.0 mg, 84%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=528.3.
To a solution of 3-[2-[2-(dimethylamino)ethoxy]ethyl]-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (150.0 mg, 0.28 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL). The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. To the above mixture were added ACN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 39% B to 49% B in 7 min; 254 nm) to afford 3-[2-[2-(dimethylamino)ethoxy]ethyl]-1-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]urea (41.0 mg, 36%) as a white solid. LCMS (ESI, m/z): [M+H]+=398.2. 1H NMR (300 MHz, DMSO-d6): δ 11.63 (s, 1H), 9.23 (s, 1H), 8.72 (s, 1H), 7.91 (d, J=8.7 Hz, 1H), 7.53-7.47 (m, 2H), 7.29-7.23 (m, 1H), 7.10 (d, J=7.5 Hz, 1H), 7.04-6.99 (m, 2H), 3.81 (s, 3H), 3.58-3.51 (m, 4H), 3.41-3.34 (m, 2H), 2.52-2.44 (m, 2H), 2.16 (s, 6H).
To a solution of 3-bromo-5-fluoro-2,4-dimethoxypyridine (345.0 mg, 1.46 mmol) in 1,4-dioxane (7.0 mL) and H2O (1.4 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (597.5 mg, 1.46 mmol), K2CO3 (404.0 mg, 2.92 mmol) and Pd(dppf)Cl2 (107.0 mg, 0.15 mmol) at room temperature under N2. The mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/EtOAc (10/1, v/v) to 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-fluoro-2,4-dimethoxypyridine (350.0 mg, 57%) as an orange solid. LCMS (ESI, m/z): [M+H]+=438.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-fluoro-2,4-dimethoxypyridine (300.0 mg, 0.69 mmol) in t-BuOH (10.0 mL) was added (1R,2R)-2-fluorocyclopropane-1-carboxamide (353.1 mg, 3.43 mmol), K2CO3 (284.0 mg, 2.06 mmol), XPhos (65.3 mg, 0.14 mmol) and Pd(OAc)2 (15.4 mg, 0.07 mmol) at room temperature under N2. The mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/EtOAc (2/1, v/v) to afford (1R,2R)-2-fluoro-N-[3-(5-fluoro-2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (196.0 mg, 64%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=505.2.
To a solution of (1R,2R)-2-fluoro-N-[3-(5-fluoro-2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (196.0 mg, 0.39 mmol) in CH2Cl2 (6.0 mL) was added TFA (3.0 mL) at room temperature. The reaction mixture was stirred at room temperature for 4 h. The mixture was concentrated under vacuum. The residue was dissolved in CH3CN (10.0 mL) and NH3·H2O (10.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions: Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 56% B in 9 min; 254 nm; RT1:8.5 min to afford (1R,2R)-2-fluoro-N-[3-(5-fluoro-2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (42.0 mg, 29%) as a white solid. LCMS (ESI, m/z): [M+H]+=375.0. 1H NMR (300 MHz, DMSO-d6): δ 11.70 (d, J=1.8 Hz, 1H), 10.64 (s, 1H), 8.12 (d, J=3.0 Hz, 1H), 7.87 (d, J=8.7 Hz, 1H), 7.71 (d, J=8.7 Hz, 1H), 7.47 (d, J=2.4 Hz, 1H), 5.05-4.80 (m, 1H), 3.81-3.80 (m, 6H), 2.25-2.21 (m, 1H), 1.70-1.60 (m, 1H), 1.19-1.14 (m, 1H).
To a solution of 3-bromo-6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (500.0 mg, 1.38 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added (2-(2,2,2-trifluoroethoxy)pyridin-3-yl)boronic acid (305.4 mg, 1.38 mmol), K2CO3 (573.1 mg, 4.15 mmol) and Pd(dppf)Cl2 (101.1 mg, 0.14 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to 6-chloro-3-(2-(2,2,2-trifluoroethoxy)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (270.0 mg, 42%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=458.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-(2,2,2-trifluoroethoxy)pyridine (126.0 mg, 0.28 mmol) in 1,4-dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (141.8 mg, 1.38 mmol), K2CO3 (268.8 mg, 1.95 mmol), BrettPhos (29.5 mg, 0.06 mmol) and Pd2(dba)3 (25.2 mg, 0.03 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford (1S,2S)-2-fluoro-N-[3-[2-(2,2,2-trifluoroethoxy)pyridin-3-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (115.0 mg, 79%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=525.2.
To a solution of (1S,2S)-2-fluoro-N-[3-[2-(2,2,2-trifluoroethoxy)pyridin-3-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (110.0 mg, 0.21 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature under N2. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in ACN (2.0 mL) and NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 46% B to 53% B in 8 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-(2-(2,2,2-trifluoroethoxy)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (12.8 mg, 15%) as a white solid. LCMS (ESI, m/z): [M+H]+=395.2. 1H NMR (400 MHz, DMSO-d6): δ 11.80 (s, 1H), 10.71 (s, 1H), 8.18 (d, J=8.8 Hz, 1H), 8.14-8.10 (m, 2H), 7.94 (d, J=8.4 Hz, 1H), 7.72 (d, J=2.8 Hz, 1H), 7.23-7.20 (m, 1H), 5.10-5.05 (m, 2H), 5.02-4.84 (m, 1H), 2.25-2.19 (m, 1H), 1.69-1.62 (m, 1H), 1.19-1.16 (m, 1H).
To a solution of 2-bromo-3-fluorophenol (2.5 g, 13.09 mmol) in DMF (50.0 mL) was added bromocyclopropane (9.2 g, 75.89 mmol) and K2CO3 (8.7 g, 62.83 mmol) at room temperature. The mixture was stirred at 180° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether to afford 2-bromo-1-cyclopropoxy-3-fluorobenzene (1.4 g, 46%) as a yellow oil.
To a solution of 2-bromo-1-cyclopropoxy-3-fluorobenzene (850.0 mg, 3.68 mmol) in dioxane (17.0 mL) and H2O (3.4 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (1503.8 mg, 3.68 mmol), K2CO3 (1016.8 mg, 7.36 mmol), XPhos (350.7 mg, 0.74 mmol) and XPhos Pd G3 (311.4 mg, 0.37 mmol) at room temperature under N2. The mixture was stirred at 70° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/EtOAc (10/1, v/v) to afford 6-chloro-3-(2-cyclopropoxy-6-fluorophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (453.0 mg, 28%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=433.1.
To a solution of 6-chloro-3-(2-cyclopropoxy-6-fluorophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (390.0 mg, 0.90 mmol) in t-BuOH (8.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (464.3 mg, 4.50 mmol), K2CO3 (311.2 mg, 2.25 mmol), Pd(OAc)2 (20.2 mg, 0.09 mmol) and XPhos (85.9 mg, 0.18 mmol) at room temperature under N2. The mixture was stirred at 100° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/EtOAc (2/1, v/v) to afford (1S,2S)—N-[3-(2-cyclopropoxy-6-fluorophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (76.0 mg, 16%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=500.2.
To a solution of (1S,2S)—N-[3-(2-cyclopropoxy-6-fluorophenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (76.0 mg, 0.15 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The reaction mixture was stirred at room temperature for 4 h. The mixture was concentrated under vacuum. The residue was dissolved in CH3CN (4.0 mL) and NH3·H2O (4.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions: Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 46% B to 56% B in 8 min, 254/220 nm to afford (1S,2S)—N-[3-(2-cyclopropoxy-6-fluorophenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (8.3 mg, 14%) as a white solid. LCMS (ESI, m/z): [M+H]+=370.0. 1H NMR (300 MHz, DMSO-d6): δ 11.66 (s, 1H), 10.64 (s, 1H), 7.87 (d, J=8.7 Hz, 1H), 7.67-7.63 (m, 1H), 7.43-7.25 (m, 3H), 6.97-6.93 (m, 1H), 5.06-4.79 (m, 1H), 3.90-3.83 (m, 1H), 2.27-2.19 (m, 1H), 1.72-1.61 (m, 1H), 1.21-1.10 (m, 1H), 0.80-0.76 (m, 2H), 0.65-0.60 (m, 2H).
To a solution of 5-bromo-4-methoxypyrimidin-2-amine (500.0 mg, 2.45 mmol) in EtOH/H2O (10.0 mL/2.5 mL) was added 2-chloroacetaldehyde (1.9 g, 9.80 mmol) and NaHCO3 (247.1 mg, 2.94 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (84/16, v/v) to afford 6-bromo-7-methoxyimidazo[1,2-a]pyrimidine (80.0 mg, 14%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=228.0.
To a solution of 6-bromo-7-methoxyimidazo[1,2-a]pyrimidine (140.0 mg, 0.61 mmol) in 1,4-dioxane/H2O (10.0 mL/2.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (200.5 mg, 0.61 mmol), K2CO3 (254.5 mg, 1.84 mmol) and Pd(dppf)Cl2 (44.9 mg, 0.06 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (9/91, v/v) to afford 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-7-methoxyimidazo[1,2-a]pyrimidine (200.0 mg, 76%) as a brown solid. LCMS (ESI, m/z): [M+H]+=430.1.
To a solution of 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-7-methoxyimidazo[1,2-a]pyrimidine (160.0 mg, 0.37 mmol) in 1,4-dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (191.8 mg, 1.86 mmol), K2CO3 (154.3 mg, 1.12 mmol), BrettPhos (40.0 mg, 0.07 mmol) and Pd2(dba)3 (34.1 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 3 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (30/70, v/v) to afford (1S,2S)-2-fluoro-N-(3-(7-methoxyimidazo[1,2-a]pyrimidin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (238.0 mg, 90%) as a brown solid. LCMS (ESI, m/z): [M+H]+=497.2.
To a solution of (1S,2S)-2-fluoro-N-(3-(7-methoxyimidazo[1,2-a]pyrimidin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (238.0 mg, 0.48 mmol) in CH2Cl2 (10.0 mL) was added TFA (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (10.0 mL) was added NH3·H2O (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 21% B to 31% B in 9 min, 254 nm) to afford (1S,2S)-2-fluoro-N-(3-(7-methoxyimidazo[1,2-a]pyrimidin-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (4.2 mg, 1%) as a white solid. LCMS (ESI, m/z): [M+H]+=367.2. 1H NMR (300 MHz, DMSO-d6): δ 11.80 (s, 1H), 10.70 (s, 1H), 9.03 (s, 1H), 8.19 (d, J=8.7 Hz, 1H), 7.95 (d, J=8.7 Hz, 1H), 7.72 (d, J=1.5 Hz, 2H), 7.43 (d, J=1.5 Hz, 1H), 5.06-4.80 (m, 1H), 4.03 (s, 3H), 2.27-2.22 (m, 1H), 1.73-1.60 (m, 1H), 1.24-1.11 (m, 1H).
To a solution of 3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (100.0 mg, 0.25 mmol) in CH2Cl2 (5.0 mL) was added phenyl chloroformate (58.8 mg, 0.38 mmol) and pyridine (80.0 mg, 1.01 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. The resulting mixture was concentrated under reduced pressure. To the above mixture in pyridine (5.0 mL) was added tert-butyl 3-(aminomethyl)azetidine-1-carboxylate (186.5 mg, 1.00 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (90/10, v/v) to afford tert-butyl 3-((3-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)ureido)methyl)azetidine-1-carboxylate (97.0 mg, 63%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=612.3.
To a solution of tert-butyl 3-((3-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)ureido)methyl)azetidine-1-carboxylate (120.0 mg, 0.20 mmol) in CH2Cl2 (1.0 mL) was added TFA (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (1.0 mL) was added NH3·H2O (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 28% B to 35% B in 8 min; 254 nm) to afford 1-(azetidin-3-ylmethyl)-3-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)urea (13.0 mg, 17.4%) as a white solid. LCMS (ESI, m/z): [M+H]+=382.4. 1H NMR (400 MHz, DMSO-d6+D2O): δ 7.46 (d, J=8.4 Hz, 1H), 7.27-7.21 (m, 1H), 7.17 (s, 1H), 6.87 (d, J=8.4 Hz, 1H), 6.73 (d, J=8.4 Hz, 2H), 3.86-3.74 (m, 2H), 3.68-3.57 (m, 2H), 3.49-3.39 (m, 2H), 2.95-2.92 (m, 1H).
To a solution of 3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (130.0 mg, 0.33 mmol) in DCM (5.0 mL) was added pyridine (104.23 mg, 1.30 mmol) and phenyl chloroformate (61.1 mg, 0.39 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the above mixture was added tert-butyl N-(3-aminopropyl)-N-methylcarbamate (183.8 mg, 0.98 mmol) and pyridine (5.0 mL) at room temperature. The resulting mixture was stirred at 60° C. for another 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to afford tert-butyl N-[3-([[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)propyl]-N-methylcarbamate (180.0 mg, 90%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=614.3.
To a solution of tert-butyl N-[3-([[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)propyl]-N-methylcarbamate (160.0 mg, 0.26 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in CH3CN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 15% B to 45% B in 9 min; 254 nm) to afford 1-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(methylamino)propyl]urea (29.9 mg, 29%) as a white solid. LCMS (ESI, m/z): [M+H]+=384.3. 1H NMR (300 MHz, DMSO-d6): δ 11.39 (s, 1H), 9.13 (s, 1H), 8.67 (s, 1H), 7.47 (d, J=8.4 Hz, 1H), 7.29-7.24 (m, 1H), 7.17 (s, 1H), 6.90 (d, J=8.4 Hz, 1H), 6.74 (d, J=8.4 Hz, 2H), 3.68 (s, 6H), 3.34-3.24 (m, 3H), 2.58-2.50 (m, 2H), 2.30 (s, 3H), 1.71-1.62 (m, 2H).
To a solution of 3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (130.0 mg, 0.33 mmol) in DCM (6.0 mL) was added pyridine (102.9 mg, 1.30 mmol) and phenyl chloroformate (61.1 mg, 0.39 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the above mixture was added dimethylaminopropylamine (99.7 mg, 0.98 mmol) and pyridine (6.0 mL) at room temperature. The resulting mixture was stirred at 60° C. for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (9/1, v/v) to afford 1-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)propyl]urea (166.0 mg, 96%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=528.3.
To a solution of 1-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)propyl]urea (172.0 mg, 0.33 mmol) in CH2Cl2 (6.0 mL) was added TFA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (6.0 mL) was added NH3·H2O (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 27% B to 57% B in 7 min; 254 nm) to afford 1-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)propyl]urea (25.8 mg, 19%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=398.3. 1H NMR (400 MHz, DMSO-d6): δ 11.35 (s, 1H), 9.11 (s, 1H), 8.60 (s, 1H), 7.46 (d, J=8.4 Hz, 1H), 7.28-7.24 (m, 1H), 7.15 (d, J=2.4 Hz, 1H), 6.91 (d, J=8.4 Hz, 1H), 6.74 (d, J=8.4 Hz, 2H), 3.68 (s, 6H), 3.26-3.21 (m, 2H), 2.29-2.25 (m, 2H), 2.14 (s, 6H), 1.68-1.61 (m, 2H).
To a solution of 3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (130.0 mg, 0.33 mmol) in DCM (13.0 mL) was added pyridine (104.2 mg, 1.30 mmol) and phenyl chloroformate (61.1 mg, 0.39 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. To the residue in pyridine (10.0 mL) was added (2-aminoethyl)dimethylamine (143.4 mg, 1.63 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 4 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) to afford 1-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-3-[2-(dimethylamino)ethyl]urea (127.0 mg, 76%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=514.3.
To a solution of 1-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-3-[2-(dimethylamino)ethyl]urea (107.0 mg, 0.21 mmol) in CH2Cl2 (8.0 mL) was added TFA (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (8.0 mL) was added NH3·H2O (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 35% B to 43% B in 8 min; 254 nm) to afford 1-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[2-(dimethylamino)ethyl]urea (26.4 mg, 33%) as a light green solid. LCMS (ESI, m/z): [M+H]+=384.2. 1H NMR (400 MHz, DMSO-d6): δ 11.28 (d, J=1.6 Hz, 1H), 9.11 (s, 1H), 8.46 (s, 1H), 7.46 (d, J=8.4 Hz, 1H), 7.28-7.24 (m, 1H), 7.15 (d, J=2.4 Hz, 1H), 6.97 (d, J=8.4 Hz, 1H), 6.73 (d, J=8.4 Hz, 2H), 3.68 (s, 6H), 3.33-3.28 (m, 2H), 2.41-2.38 (m, 2H), 2.21 (s, 6H).
To a solution of 2-fluoro-3-methoxyaniline (2.0 g, 14.17 mmol) in DMF (20.0 mL) was added NBS (2.5 g, 14.17 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was quenched with Na2SO3 (aq.). The resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 4-bromo-2-fluoro-3-methoxyaniline (2.1 g, 67%) as a white solid. LCMS (ESI, m/z): [M+H]+=220.0.
To a mixture of 4-bromo-2-fluoro-3-methoxyaniline (1.0 g, 4.54 mmol) and NH4SCN (380.5 mg, 5.00 mmol) in acetic acid (40.0 mL) was added dropwise Br2 (871.5 mg, 5.45 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the mixture was basified to pH=8 with saturated Na2CO3 (aq.). The resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 6-bromo-4-fluoro-5-methoxy-1,3-benzothiazol-2-amine (750.0 mg, 59%) as an off-white solid. LCMS (ESI, m/z): [M+H]+=276.9.
To a mixture of 6-bromo-4-fluoro-5-methoxy-1,3-benzothiazol-2-amine (380.0 mg, 1.37 mmol) and 2-methyl-2-propylnitrite (212.1 mg, 2.05 mmol) in THE (10.0 mL) was added dropwise DMSO (10.7 mg, 0.14 mmol). The resulting mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 6-bromo-4-fluoro-5-methoxy-1,3-benzothiazole (200.0 mg, 55%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=261.9.
To a mixture of 6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-ylboronic acid (548.4 mg, 1.68 mmol) and 6-bromo-4-fluoro-5-methoxy-1,3-benzothiazole (400.0 mg, 1.52 mmol) in dioxane (10.0 mL) were added [AMPhosPdCl2]2 (216.1 mg, 0.31 mmol), Cs2CO3 (1491.7 mg, 4.58 mmol) and H2O (1.0 mL) at room temperature. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 6-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-fluoro-5-methoxy-1,3-benzothiazole (400.0 mg, 56%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=464.1.
To a mixture of 6-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-fluoro-5-methoxy-1,3-benzothiazole (300.0 mg, 0.68 mmol) and (1S,2S)-2-fluorocyclopropane-1-carboxamide (333.3 mg, 3.23 mmol) in 1,4-dioxane (5.0 mL) was added Pd2(dba)3 (118.4 mg, 0.13 mmol), BrettPhos (138.8 mg, 0.26 mmol) and K2CO3 (268.0 mg, 1.94 mmol). The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(4-fluoro-5-methoxy-1,3-benzothiazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (50.0 mg, 14%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=531.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(4-fluoro-5-methoxy-1,3-benzothiazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (45.0 mg, 0.09 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 35% B in 7 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(4-fluoro-5-methoxy-1,3-benzothiazol-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (11.6 mg, 34%) as a white solid. LCMS (ESI, m/z): [M+H]+=401.1. 1H NMR (300 MHz, DMSO-d6): δ 11.80 (s, 1H), 10.68 (s, 1H), 9.36 (s, 1H), 8.19-8.17 (m, 2H), 7.93 (d, J=8.4 Hz, 1H), 7.75 (d, J=2.7 Hz, 1H), 5.05-4.78 (m, 1H), 3.74 (s, 3H), 2.25-2.18 (m, 1H), 1.88-1.58 (m, 1H), 1.22-1.07 (m, 1H).
To a solution of 5-bromo-6-methoxypyridin-2-amine (1.5 g, 7.42 mmol) in DMF (10.0 mL) was added DMF-DMA (972.0 mg, 8.17 mmol). The resulting mixture was stirred at 70° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (E)-N′-(5-bromo-6-methoxypyridin-2-yl)-N,N-dimethylformimidamide (1.5 g, 79%) as an off-white solid. LCMS (ESI, m/z): [M+H]+=258.0.
To a solution of (E)-N′-(5-bromo-6-methoxypyridin-2-yl)-N,N-dimethylformimidamide (1.5 g, 5.81 mmol) in methanol (10.0 mL) was added hydroxylamine hydrochloride (1.5 g, 21.44 mmol) and NaOAc (1.5 g, 18.85 mmol). The resulting mixture was stirred at 70° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (E)-N′-(5-bromo-6-methoxypyridin-2-yl)-N-hydroxymethanimidamide (1.4 g, 98%) as an off-white solid. LCMS (ESI, m/z): [M+H]+=246.0.
To a solution of (E)-N′-(5-bromo-6-methoxypyridin-2-yl)-N-hydroxymethanimidamide (1.4 g, 5.69 mmol) in DCM (10.0 mL) was added TFAA (1.3 g, 6.27 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 6-bromo-5-methoxy-[1,2,4]triazolo[1,5-a]pyridine (600.0 mg, 46%) as a white solid. LCMS (ESI, m/z): [M+H]+=228.0.
To a solution of 6-bromo-5-methoxy-[1,2,4]triazolo[1,5-a]pyridine (300.0 mg, 1.36 mmol) in dioxane (5.0 mL) and H2O (0.5 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (645.5 mg, 1.59 mmol), (AMPhospdCl2)2 (93.1 mg, 0.12 mmol) and K3PO4 (837.7 mg, 3.94 mmol). The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 6-chloro-3-[5-methoxy-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (200.0 mg, 35%) as a white solid. LCMS (ESI, m/z): [M+H]+=430.1.
To a solution of 6-chloro-3-[5-methoxy-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (200.0 mg, 0.45 mmol) in t-BuOH (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (479.4 mg, 4.61 mmol), Pd(OAc)2 (20.9 mg, 0.09 mmol), XPhos (22.7 mg, 0.05 mmol) and K2CO3 (192.8 mg, 1.39 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)-2-fluoro-N-(3-(5-methoxy-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (70.0 mg, 30%) as a white solid. LCMS (ESI, m/z): [M+H]+=497.2.
To a solution of (1S,2S)-2-fluoro-N-(3-[5-methoxy-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (70.0 mg, 0.14 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 24% B to 32% B in 7 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-[5-methoxy-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (1.2 mg, 2%) LCMS (ESI, m/z): [M+H]+=367.3. 1H NMR (300 MHz, DMSO-d6): δ 11.87 (s, 1H), 10.70 (s, 1H), 8.55 (s, 1H), 8.09 (d, J=8.4 Hz, 1H), 7.99-7.93 (m, 2H), 7.73-7.70 (m, 2H), 5.05-4.82 (m, 1H), 4.03 (s, 3H), 2.30-2.19 (m, 1H), 1.70-1.60 (m, 1H), 1.23-1.13 (m, 1H).
To a solution of 3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (110.0 mg, 0.28 mmol) in CH2Cl2 (3.0 mL) was added pyridine (87.1 mg, 1.10 mmol) and phenyl chloroformate (51.7 mg, 0.33 mmol) at 0° C. The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. To the above mixture was added pyridine (2.0 mL) and tert-butyl 4-(2-aminoethyl)piperazine-1-carboxylate (189.4 mg, 0.83 mmol). The resulting mixture was stirred at 60° C. for 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (95/5, v/v) to afford tert-butyl 4-[2-([[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (110.0 mg, 61%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=655.4.
To a solution of tert-butyl 4-[2-([[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (120.0 mg, 0.18 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL). The mixture was stirred at room temperature for 4 h. The mixture was concentrated under vacuum. To the above residue were added ACN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 32% B to 42% B in 8 min; 254 nm) to afford 3-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(piperazin-1-yl)ethyl]urea (24.6 mg, 31%) as a white solid. LCMS (ESI, m/z): [M+H]+=425.3. 1H NMR (300 MHz, DMSO-d6): δ 11.30 (s, 1H), 9.15 (s, 1H), 8.41 (s, 1H), 7.46 (d, J=8.7 Hz, 1H), 7.29-7.24 (m, 1H), 7.17 (d, J=2.1 Hz, 1H), 6.99 (d, J=8.4 Hz, 1H), 6.74 (d, J=8.4 Hz, 2H), 3.69 (s, 6H), 2.73-2.70 (m, 4H), 2.46-2.37 (m, 6H).
To a solution of 6-chloro-3-(2-cyclopropoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (320.0 mg, 0.77 mmol) in THE (5.0 mL) was added X-Phos (73.3 mg, 0.15 mmol), Pd2(dba)3 (70.4 mg, 0.08 mmol) and LiHMDS (1.2 mL, 1 mol/L) at room temperature. The resulting mixture was stirred at 60° C. for 1 h. After the reaction was completed, the reaction was quenched with water. The resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (93/7, v/v) to afford 3-(2-cyclopropoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-amine (120.0 mg, 39%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=397.2.
To a solution of 3-(2-cyclopropoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-amine (100.0 mg, 0.25 mmol) in CH2Cl2 (3.0 mL) was added pyridine (79.8 mg, 1.01 mmol) and phenyl chloroformate (47.4 mg, 0.30 mmol) at 0° C. The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. To the above residue was added pyridine (2.0 mL) and tert-butyl 4-(2-aminoethyl)piperazine-1-carboxylate (173.5 mg, 0.76 mmol). The resulting mixture was stirred at 60° C. for 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (94/6, v/v) to afford tert-butyl 4-[2-([[3-(2-cyclopropoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (100.0 mg, 60%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=652.4.
To a solution of tert-butyl 4-[2-([[3-(2-cyclopropoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (80.0 mg, 0.12 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL). The mixture was stirred at room temperature for 3 h. The mixture was concentrated under vacuum. To the above residue were added ACN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 3 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column, 19×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 47% B to 68% B in 9 min; 254 nm) to afford 3-[3-(2-cyclopropoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-6-yl]-1-[2-(piperazin-1-yl)ethyl]urea (14.0 mg, 27%) as a white solid. LCMS (ESI, m/z): [M+H]+=422.3. 1H NMR (300 MHz, DMSO-d6): δ 13.30 (s, 1H), 9.61 (s, 1H), 8.43-8.39 (m, 1H), 7.90 (d, J=8.7 Hz, 1H), 7.58 (d, J=1.2 Hz, 1H), 7.49-7.41 (m, 2H), 7.16-7.06 (m, 2H), 3.93-3.87 (m, 1H), 3.37-3.23 (s, 3H), 2.78-2.75 (m, 4H), 2.47-2.39 (m, 6H), 0.83-0.73 (m, 2H), 0.68-0.63 (m, 2H).
To a mixture of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2,4-dimethoxypyridine (600.0 mg, 1.43 mmol), Cs2CO3 (1.4 g, 4.28 mmol) and BocNH2 (836.8 mg, 7.14 mmol) in dioxane (5.0 mL) were added Pd(OAc)2 (64.1 mg, 0.28 mmol) and X-Phos (272.4 mg, 0.57 mmol). The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford tert-butyl N-[3-(2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamate (550.0 mg, 77%) as a white solid. LCMS (ESI, m/z): [M+H]+=501.2.
A mixture of tert-butyl N-[3-(2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamate (250.0 mg, 0.5 mmol) and formic acid (10.0 mL) in DCM (20.0 mL) was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O. The pH value of the mixture was adjusted to 8 with saturated NaHCO3 solution and then extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 3-(2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (200.0 mg, crude) as a yellow oil. LCMS (ESI, m/z): [M+H]+=401.2.
To a mixture of 3-(2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (100.0 mg, 0.25 mmol) and pyridine (79.9 mg, 1.0 mmol) in DCM (10.0 mL) was added phenyl chloroformate (46.9 mg, 0.30 mmol). The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in pyridine (5.0 mL) was added tert-butyl 4-(2-aminoethyl)piperazine-1-carboxylate (343.5 mg, 1.50 mmol) at room temperature. The resulting mixture was stirred at 60° C. for another 16 h. After the reaction was completed, the mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford tert-butyl 4-[2-([[3-(2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (100.0 mg, 61%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=656.4.
To a solution of tert-butyl 4-[2-([[3-(2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (170.0 mg, 0.26 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 24% B to 32% B in 7 min; 254 nm) to afford 3-[3-(2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(piperazin-1-yl)ethyl]urea (13.2 mg, 11%) as a white solid. LCMS (ESI, m/z): [M+H]+=426.2. 1H NMR (300 MHz, CDCl3) δ 9.74 (s, 1H), 9.58 (s, 1H), 8.09 (d, J=5.7 Hz, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.33 (s, 1H), 7.21 (s, 1H), 6.66 (d, J=5.7 Hz, 1H), 6.48-6.44 (m, 1H), 3.93 (s, 3H), 3.84 (s, 3H), 3.61-3.56 (m, 2H), 3.07-3.02 (m, 4H), 2.76-2.64 (m, 6H).
To a solution of 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.22 mmol) in 1,4-dioxane/H2O (20.0 mL/5.0 mL) was added 5-bromo-4,6-dimethoxypyrimidine (267.9 mg, 1.22 mmol), K2CO3 (507.1 mg, 3.67 mmol) and Pd(dppf)Cl2 (89.4 mg, 0.12 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4,6-dimethoxypyrimidine (240.0 mg, 46%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=421.1.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4,6-dimethoxypyrimidine (110.0 mg, 0.26 mmol) in THF (8.0 mL) was added XPhos (24.9 mg, 0.05 mmol), Pd2(dba)3 (15.0 mg, 0.03 mmol) and LiHMDS (0.4 mL, 1 mol/L) at room temperature under N2. The resulting mixture was stirred at 65° C. for 0.5 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 3-(4,6-dimethoxypyrimidin-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (100.0 mg, 95%) as a brown yellow solid. LCMS (ESI, m/z): [M+H]+=402.2.
To a solution of 3-(4,6-dimethoxypyrimidin-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (180.0 mg, 0.45 mmol) in DCM (10.0 mL) was added pyridine (143.6 mg, 1.79 mmol) and phenyl chloroformate (84.2 mg, 0.54 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added tert-butyl 4-(2-aminoethyl)piperazine-1-carboxylate (308.4 mg, 1.35 mmol) and pyridine (10.0 mL) at room temperature. The resulting mixture was stirred at 60° C. for another 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with DCM/MeOH (20/1, v/v) to afford tert-butyl 4-[2-([[3-(4,6-dimethoxypyrimidin-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (200.0 mg, 67%) as a white solid. LCMS (ESI, m/z): [M+H]+=657.3.
To a solution of tert-butyl 4-[2-([[3-(4,6-dimethoxypyrimidin-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (180.0 mg, 0.27 mmol) in DCM (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (3.0 mL) was added NH3·H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 24% B to 34% B in 7 min, 254/220 nm) to afford 3-[3-(4,6-dimethoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(piperazin-1-yl)ethyl]urea (33.5 mg, 28%) as a white solid. LCMS (ESI, m/z): [M+H]+=427.3. 1H NMR (400 MHz, DMSO-d6): δ 11.53 (s, 1H), 9.20 (s, 1H), 8.46 (s, 1H), 8.32 (s, 1H), 7.62 (d, J=8.8 Hz, 1H), 7.37 (s, 1H), 7.06 (d, J=8.8 Hz, 1H), 3.91 (s, 6H), 3.39-3.30 (m, 3H), 2.71 (s, 4H), 2.51-2.50 (m, 2H), 2.45-2.37 (m, 4H).
To a solution of 6-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-methoxy-1,3-benzothiazole (380.0 mg, 0.85 mmol) in 1.4-dioxane (10.0 mL) was added tert-butyl carbamate (139.7 mg, 1.19 mmol), Xphos (81.2 mg, 0.17 mmol), Cs2CO3 (694.0 mg, 2.13 mmol) and Pd(OAc)2 (19.1 mg, 0.09 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (7/3, v/v) to afford tert-butyl N-[3-(5-methoxy-1,3-benzothiazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamate (420.0 mg, 93%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=527.2.
To a solution of tert-butyl N-[3-(5-methoxy-1,3-benzothiazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamate (470.0 mg, 0.89 mmol) in CH2Cl2 (10.0 mL) was added formic acid (FA, 10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the pH value of the mixture was adjusted to 8 with NaHCO3 solution. The resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 3-(5-methoxy-1,3-benzothiazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (380.0 mg, crude) as a brown oil. LCMS (ESI, m/z): [M+H]+=427.2.
To a solution of 3-(5-methoxy-1,3-benzothiazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (290.0 mg, crude) in DCM (8.0 mL) was added pyridine (215.1 mg, 2.72 mmol) and phenyl chloroformate (255.4 mg, 1.63 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. To the above mixture was added tert-butyl 4-(2-aminoethyl)piperazine-1-carboxylate (935.3 mg, 4.08 mmol) and pyridine (8.0 mL) at room temperature. The resulting mixture was stirred at 60° C. for another 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (15/1, v/v) to afford tert-butyl 4-[2-([[3-(5-methoxy-1,3-benzothiazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (155.0 mg, 33%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=682.3.
To a solution of tert-butyl 4-[2-([[3-(5-methoxy-1,3-benzothiazol-6-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (155.0 mg, 0.23 mmol) in CH2Cl2 (8.0 mL) was added TFA (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (8.0 mL) was added NH3·H2O (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 7% B to 20% B in 8 min; 254/220 nm) to afford 3-[3-(5-methoxy-1,3-benzothiazol-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(piperazin-1-yl)ethyl]urea; bis(formic acid) (12.9 mg, 9%) as a white semi-solid. LCMS (ESI, m/z): [M+H]+=452.3. 1H NMR (400 MHz, DMSO-d6): δ 11.58 (s, 1H), 9.31 (s, 1H), 9.23 (s, 1H), 8.41 (s, 1H), 8.30 (d, J=3.6 Hz, 2H), 8.24 (s, 1H), 8.00 (d, J=8.4 Hz, 1H), 7.75 (s, 1H), 7.57 (d, J=2.4 Hz, 1H), 7.11 (d, J=8.8 Hz, 1H), 3.92 (s, 3H), 3.37-3.32 (m, 2H), 3.02-2.91 (m, 4H), 2.67-2.55 (m, 5H).
To a solution of 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.22 mmol) in dioxane/H2O (20.0 mL/4.0 mL) was added 5-bromo-6-methoxy-1,3-benzothiazole (298.6 mg, 1.22 mmol), K2CO3 (507.1 mg, 3.67 mmol) and Pd(dppf)Cl2 (89.5 mg, 0.12 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1,3-benzothiazole (250.0 mg, 48%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=446.1.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-6-methoxy-1,3-benzothiazole (230.0 mg, 0.52 mmol) in 1,4-dioxane (24.0 mL) was added tert-butyl carbamate (84.6 mg, 0.72 mmol), XPhos (49.2 mg, 0.11 mmol), Cs2CO3 (420.0 mg, 1.29 mmol) and Pd(OAc)2 (11.6 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with ACN/H2O (1/3, v/v) to afford tert-butyl N-[3-(6-methoxy-1,3-benzothiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamate (160.0 mg, 59%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=527.2.
To a solution of tert-butyl N-[3-(6-methoxy-1,3-benzothiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamate (140.0 mg, 0.27 mmol) in DCM (15.0 mL) was added HCOOH (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 30 min. After the reaction was completed, the reaction mixture was diluted with H2O. The pH value of the mixture was adjusted to 8 with saturated NaHCO3 solution. The mixture was extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated in vacuo to afford 3-(6-methoxy-1,3-benzothiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (110.0 mg, crude) as a yellow oil. LCMS (ESI, m/z): [M+H]+=427.2.
To a solution of 3-(6-methoxy-1,3-benzothiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (80.0 mg, crude) in DCM (8.0 mL) was added pyridine (60.1 mg, 0.75 mmol) and phenyl chloroformate (35.2 mg, 0.23 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h under N2. The resulting mixture was concentrated under reduced pressure. To the residue in pyridine (8.0 mL) was added tert-butyl 4-(2-aminoethyl)piperazine-1-carboxylate (215.0 mg, 0.94 mmol) at room temperature. The resulting mixture was stirred at 60° C. for another 4 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) to afford tert-butyl 4-[2-([[3-(6-methoxy-1,3-benzothiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (120.0 mg, 94%) as a brown solid. LCMS (ESI, m/z): [M+H]+=682.3.
To a solution of tert-butyl 4-[2-([[3-(6-methoxy-1,3-benzothiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)ethyl]piperazine-1-carboxylate (120.0 mg, 0.18 mmol) in CH2Cl2 (10.0 mL) was added TFA (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (10.0 mL) was added NH3·H2O (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 4 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 12% B to 42% B in 9 min; 254 nm) to afford 3-[3-(6-methoxy-1,3-benzothiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(piperazin-1-yl)ethyl]urea (22.4 mg, 28%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=452.3. 1H NMR (400 MHz, CD3OD): δ 9.09 (d, J=1.6 Hz, 1H), 8.16 (d, J=1.6 Hz, 1H), 8.02-8.00 (m, 1H), 7.74 (d, J=3.6 Hz, 1H), 7.55 (d, J=0.8 Hz, 1H), 6.79 (d, J=8.4 Hz, 1H), 3.96 (s, 3H), 3.57-3.49 (m, 2H), 2.94-2.92 (m, 4H), 2.79-2.60 (m, 6H).
To a solution of 5-bromo-6-methoxy-1H-pyrrolo[2,3-b]pyridine (500.0 mg, 2.20 mmol) in THE (5.0 mL) was added NaH (158.5 mg, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h under N2. Then SEM-Cl (440.5 mg, 2.64 mmol) was added to the mixture at 0° C. under N2. The resulting mixture was stirred at 0° C. for another 1 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (93/7, v/v) to afford 5-bromo-6-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (620.0 mg, 80%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=357.1.
To a solution of 5-bromo-6-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (320.0 mg, 0.90 mmol) in 1,4-dioxane/H2O (10.0/2.0 mL) was added (6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)boronic acid (366.1 mg, 0.90 mmol), K2CO3 (371.3 mg, 2.69 mmol) and Pd(dppf)Cl2 (65.5 mg, 0.09 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (93/7, v/v) to afford 6-chloro-6′-methoxy-1,1′-bis ((2-(trimethylsilyl)ethoxy)methyl)-1H,1′H-3,5′-bipyrrolo[2,3-b]pyridine (210.0 mg, 41%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=559.2.
To a solution of 6-chloro-6′-methoxy-1,1′-bis ((2-(trimethylsilyl)ethoxy)methyl)-1H,1′H-3,5′-bipyrrolo[2,3-b]pyridine (210.0 mg, 0.38 mmol) in 1,4-dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (193.6 mg, 1.87 mmol), Cs2CO3 (367.0 mg, 1.13 mmol), BrettPhos (40.3 mg, 0.08 mmol) and BrettPhos Pd G3 (34.0 mg, 0.04 mmol) at room temperature under N2, The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (83/17, v/v) to afford the mixture of (1S,2S)-2-fluoro-N-(6′-methoxy-1,1′-bis((2-(trimethylsilyl)ethoxy)methyl)-1H,1′H-[3,5′-bipyrrolo[2,3-b]pyridin]-6-yl)cyclopropane-1-carboxamide and (1R,2S)-2-fluoro-N-(6′-methoxy-1,1′-bis((2-(trimethylsilyl)ethoxy)methyl)-1H,1′H-[3,5′-bipyrrolo[2,3-b]pyridin]-6-yl)cyclopropane-1-carboxamide (110.0 mg, 49%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=626.3.
To a solution of the mixture of (1S,2S)-2-fluoro-N-(6′-methoxy-1,1′-bis((2-(trimethylsilyl)ethoxy)methyl)-1H,1′H-[3,5′-bipyrrolo[2,3-b]pyridin]-6-yl)cyclopropane-1-carboxamide and (1R,2S)-2-fluoro-N-(6′-methoxy-1,1′-bis((2-(trimethylsilyl)ethoxy)methyl)-1H,1′H-[3,5′-bipyrrolo[2,3-b]pyridin]-6-yl)cyclopropane-1-carboxamide (50.0 mg, 0.08 mmol) in DMF (2.0 mL) was added TBAF (62.7 mg, 0.24 mmol) and DEA (24.1 mg, 0.40 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (50/50, v/v) and then Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 30% B to 40% B in 11 min; 254 nm) to afford (1R,2S)-2-fluoro-N-(6′-methoxy-1H,1′H-[3,5′-bipyrrolo[2,3-b]pyridin]-6-yl)cyclopropane-1-carboxamide (3.1 mg, 10%) as a white solid. LCMS (ESI, m/z): [M+H]+=366.1. 1H NMR (300 MHz, DMSO-d6): δ 11.56-11.46 (m, 2H), 10.76 (s, 1H), 8.05-8.00 (m, 2H), 7.85 (d, J=7.8 Hz, 1H), 7.58 (s, 1H), 7.22-7.20 (m, 1H), 6.40 (s, 1H), 5.01-4.79 (m, 1H), 3.94 (s, 3H), 2.60-2.51 (m, 1H), 1.57-1.45 (m, 1H), 1.31-1.20 (m, 1H).
To a solution of 5-bromo-4,6-dimethoxy-2-methylpyrimidine (400.0 mg, 1.72 mmol) in dioxane/H2O (16.0/4.0 mL) was added (6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)boronic acid (560.6 mg, 1.72 mmol), K2CO3 (711.6 mg, 5.15 mmol) and Pd(dppf)Cl2 (125.6 mg, 0.17 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 6-chloro-3-(4,6-dimethoxy-2-methylpyrimidin-5-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (270.0 mg, 20%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=435.2.
To a solution of 6-chloro-3-(4,6-dimethoxy-2-methylpyrimidin-5-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (100.0 mg, 0.23 mmol) in 1,4-dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (118.5 mg, 1.15 mmol), K2CO3 (95.3 mg, 0.69 mmol), Brettphos (24.7 mg, 0.05 mmol) and BrettPhos Pd G3 (20.8 mg, 0.02 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)—N-(3-(4,6-dimethoxy-2-methylpyrimidin-5-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (60.0 mg, 52%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=502.2.
To a solution of (1S,2S)—N-(3-(4,6-dimethoxy-2-methylpyrimidin-5-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (50.0 mg, 0.10 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 36% to 46% in 8 min; 254 nm) to afford (1S,2S)—N-(3-(4,6-dimethoxy-2-methylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (13.6 mg, 36%) as a white solid. LCMS (ESI, m/z): [M+H]+=372.2. 1H NMR (300 MHz, DMSO-d6): δ 11.59 (s, 1H), 10.59 (s, 1H), 7.84 (d, J=8.4 Hz, 1H), 7.68 (d, J=8.7 Hz, 1H), 7.42 (d, J=2.7 Hz, 1H), 5.05-4.78 (m, 1H), 3.88 (s, 6H), 2.52 (s, 3H), 2.27-2.20 (m, 1H), 1.72-1.60 (m, 1H), 1.25-1.08 (m, 1H).
To a mixture of 6-chloro-3-[5-methoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (300.0 mg, 0.67 mmol), BocNH2 (235.8 mg, 2.01 mmol) and Cs2CO3 (655.9 mg, 2.01 mmol) in dioxane (5.0 mL) were added Pd(OAc)2 (15.1 mg, 0.07 mmol) and X-Phos (63.9 mg, 0.14 mmol). The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford tert-butyl N-(3-[5-methoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)carbamate (330.0 mg, 93%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=528.2.
A mixture of tert-butyl N-(3-[5-methoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)carbamate (300.0 mg, 0.57 mmol) and formic acid (10.0 mL) in DCM (20.0 mL) was stirred at room temperature for 16 h. After the reaction was completed, the pH value of the mixture was adjusted to 7 with NaHCO3 solution. The resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 3-(5-methoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (200.0 mg, crude) as a yellow oil. LCMS (ESI, m/z): [M+H]+=428.1.
To a mixture of 3-[5-methoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (100.0 mg, 0.23 mmol) and pyridine (74.0 mg, 0.94 mmol) in DCM (2.0 mL) was added phenyl chloroformate (73.2 mg, 0.46 mmol). The resulting mixture was stirred at room temperature for 3 h. The mixture was evaporated in vacuo. To the above residue was added pyridine (4.0 mL) and 2-(4-ethylpiperazin-1-yl)ethanamine (183.9 mg, 1.17 mmol). The resulting mixture was stirred at 60° C. for another 16 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/5, v/v) to afford 1-(2-(4-ethylpiperazin-1-yl)ethyl)-3-(3-(5-methoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)urea (35.0 mg, 25%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=611.3.
To a solution of 1-(2-(4-ethylpiperazin-1-yl)ethyl)-3-(3-(5-methoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)urea (35.0 mg, 0.06 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue was added CH3CN (2.0 mL) and NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 35% B in 7 min; 254 nm) to afford 1-(2-(4-ethylpiperazin-1-yl)ethyl)-3-(3-(5-methoxythiazolo[4,5-b]pyridin-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)urea (3.8 mg, 13%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=481.4. 1H NMR (400 MHz, CD3OD): δ 9.38 (s, 1H), 8.61 (s, 1H), 8.12 (d, J=8.4 Hz, 1H), 7.71 (s, 1H), 6.83 (d, J=8.4 Hz, 1H), 4.14 (s, 3H), 3.58-3.55 (m, 2H), 2.94-2.84 (m, 4H), 2.82-2.74 (m, 8H), 1.25-1.21 (m, 3H).
To a mixture of 2-chloro-6-methoxypyridine-3-carboxylic acid (5.0 g, 26.6 mmol) and NaOAc (4.3 g, 53.27 mmol) in AcOH (50.0 mL) was added dropwises Br2 (17.04 g, 106.62 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction was quenched with water at room temperature and then filtered. The solid was washed with water and petroleum ether to afford 5-bromo-2-chloro-6-methoxypyridine-3-carboxylic acid (2.0 g, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=265.9
The solution of 5-bromo-2-chloro-6-methoxypyridine-3-carboxylic acid (2.0 g, 7.50 mmol) in BH3. THE (30.0 mL, 1 mol/L) was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was quenched with MeOH and then concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford (5-bromo-2-chloro-6-methoxypyridin-3-yl)methanol (1.4 g, 73%) as a white solid. LCMS (ESI, m/z): [M+H]+=251.9.
To a solution of (5-bromo-2-chloro-6-methoxypyridin-3-yl)methanol (1.4 g, 5.54 mmol) in DCM (20.0 mL) was added Dess-Martin reagent (2.3 g, 5.54 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 5-bromo-2-chloro-6-methoxypyridine-3-carbaldehyde (1.2 g, 86%) as a white solid. LCMS (ESI, m/z): [M+H]+=249.9.
To a mixture of 5-bromo-2-chloro-6-methoxypyridine-3-carbaldehyde (400.0 mg, 1.59 mmol) and K2CO3 (882.8 mg, 6.38 mmol) in dioxane (5.0 mL) was added NH2NH2·H2O (2.3 g, 80%) at room temperature. The resulting mixture was stirred at 90° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 5-bromo-6-methoxy-1H-pyrazolo[3,4-b]pyridine (130.0 mg, 35%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=228.0.
To a solution of 5-bromo-6-methoxy-1H-pyrazolo[3,4-b]pyridine (130.0 mg, 0.57 mmol) in THF (2.0 mL) was added NaH (27.3 mg, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h under N2. Then SEM-Cl (142.5 mg, 0.85 mmol) was added to the mixture at 0° C. The resulting mixture was stirred at 0° C. for another 2 h. After the reaction was completed, the reaction mixture was quenched with saturated NH4Cl solution and then concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 5-bromo-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (200.0 mg, 97%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=358.1.
To a solution of 5-bromo-6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridine (200.0 mg, 0.55 mmol) in dioxane/H2O (5.0/0.5 mL) were added 6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-ylboronic acid (182.3 mg, 0.55 mmol), Pd(dppf)Cl2 (40.8 mg, 0.05 mmol) and K2CO3 (231.4 mg, 1.67 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford 6-chloro-3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (150.0 mg, 47%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=560.2.
To a solution of 6-chloro-3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (110.0 mg, 0.19 mmol) in BuOH (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (101.2 mg, 0.98 mmol), Pd(OAc)2 (4.4 mg, 0.02 mmol), K2CO3 (81.4 mg, 0.58 mmol) and Xphos (18.7 mg, 0.03 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (65.0 mg, 52%) as a white solid. LCMS (ESI, m/z): [M+H]+=627.3
To a solution of (1S,2S)-2-fluoro-N-[3-(6-methoxy-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrazolo[3,4-b]pyridin-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (110.0 mg, 0.17 mmol) in DCM (3.0 mL) was added TFA (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 24% B to 42% B in 9 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-[6-methoxy-1H-pyrazolo[3,4-b]pyridin-5-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl) cyclopropane-1-carboxamide (6.0 mg, 9%) as a white solid. LCMS (ESI, m/z): [M+H]+=367.1. 1H NMR (300 MHz, DMSO-d6): δ 13.36 (s, 1H), 11.63 (s, 1H), 10.65 (s, 1H), 8.26 (s, 1H), 8.08-7.90 (m, 3H), 7.63 (d, J=2.4 Hz, 1H), 5.07-4.80 (m, 1H), 4.00 (s, 3H), 2.44-2.31 (m, 1H), 1.73-1.60 (m, 1H), 1.24-1.12 (m, 1H).
To a solution of 4-fluorobenzene-1,3-diol (10.0 g, 78.06 mmol) in CH3OH (100.0 mL) was added NIS (8.8 g, 39.03 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 16. After the reaction was completed, the reaction was quenched with saturated Na2S203 solution at 0° C. The resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (93/7, v/v) to afford 4-fluoro-2-iodobenzene-1,3-diol (9.7 g, 49%) as a white solid.
To a solution of 4-fluoro-2-iodobenzene-1,3-diol (9.7 g, 38.19 mmol) in DMF (20.0 mL) was added K2CO3 (15.8 g, 114.32 mmol) and CH3I (11.9 g, 83.63 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (100/0, v/v) to afford 1-fluoro-3-iodo-2,4-dimethoxybenzene (5.8 g, 54%) as a white solid.
To a solution of 1-fluoro-3-iodo-2,4-dimethoxybenzene (2.0 g, 7.09 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (3.5 g, 8.51 mmol), K2CO3 (2.9 g, 21.27 mmol) and Pd(dppf)Cl2 (518.8 mg, 0.71 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (94/6, v/v) to afford 6-chloro-3-(3-fluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (2.3 g, 74%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=437.1.
To a solution of 6-chloro-3-(3-fluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (500.0 mg, 1.14 mmol) in 1,4-dioxane (5.0 mL) was added diphenylmethanimine (622.1 mg, 3.43 mmol), Cs2CO3 (1.1 g, 3.43 mmol), BrettPhos (122.8 mg, 0.23 mmol) and BrettPhos Pd G3 (103.7 mg, 0.11 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (56/44, v/v) to afford N-(3-(3-fluoro-2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-1,1-diphenylmethanimine (200.0 mg, 30%) as a white solid. LCMS (ESI, m/z): [M+H]+=582.3.
To a solution of N-[3-(3-fluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1,1-diphenylmethanimine (200.0 mg, 0.34 mmol) in DCM (5.0 mL) was added HCOOH (0.5 ml) at room temperature under N2. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (47/53, v/v) to afford 3-(3-fluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (100.0 mg, 69%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=418.2.
To a solution of 3-(3-fluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (100.0 mg, 0.24 mmol) in CH2Cl2 (5.0 mL) was added pyridine (75.7 mg, 0.96 mmol) and phenyl chloroformate (37.5 mg, 0.24 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. The resulting mixture was concentrated under reduced pressure. To the above mixture in pyridine (5.0 mL) was added 2-(4-methylpiperazin-1-yl)ethanamine (137.4 mg, 0.96 mmol) at 0° C. under N2. The resulting mixture was stirred at 60° C. for another 1 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (88/12, v/v) to afford 3-[3-(3-fluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (100.0 mg, 71%) as a white solid. LCMS (ESI, m/z): [M+H]+=587.3.
To a solution of 3-[3-(3-fluoro-2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (140.0 mg, 0.24 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XSelect CSH Fluoro Phenyl, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: MeOH—Preparative; Flow rate: 25 mL/min; Gradient: 63% B to 87% B in 7 min; 254 nm) to afford 3-[3-(3-fluoro-2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (27.4 mg, 25%) as a white solid. LCMS (ESI, m/z): [M+H]+=457.3. 1H NMR (400 MHz, CDCl3): δ 10.18 (s, 1H), 9.78 (s, 1H), 7.70 (d, J=8.4 Hz, 1H), 7.33 (d, J=2.4 Hz, 1H), 7.28 (s, 1H), 7.08-7.03 (m, 1H), 6.69-6.66 (m, 1H), 6.43 (d, J=8.4 Hz, 1H), 3.79 (s, 3H), 3.64-3.61 (m, 5H), 2.90-2.78 (m, 6H), 2.64-2.61 (m, 4H), 2.41 (s, 3H).
To a solution of 5-bromo-6-methoxypyridin-2-amine (3.0 g, 14.77 mmol) in acetone (40.0 mL) was added benzoyl isothiocyanate (2.9 g, 17.73 mmol) at room temperature under N2. The resulting mixture was stirred at 60° C. for 30 min. After the reaction was completed, the resulting mixture was concentrated under reduced pressure to afford 1-benzoyl-3-(5-bromo-6-methoxypyridin-2-yl)thiourea (5.7 g, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=366.0.
To a solution of 1-benzoyl-3-(5-bromo-6-methoxypyridin-2-yl)thiourea (5.2 g, crude) in MeOH/H2O (20.0/10.0 mL) was added NaOH (1.1 g, 28.54 mmol) at room temperature under N2. The resulting mixture was stirred at 60° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 5-bromo-6-methoxypyridin-2-ylthiourea (3.5 g, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=262.0.
To a solution of 5-bromo-6-methoxypyridin-2-ylthiourea (1.5 g, crude) in CHCl3 (15.0 mL) was added Br2 (0.9 g, 5.72 mmol) at 0° C. under N2. The resulting mixture was stirred at 60° C. for 16 h. After the reaction was completed, the reaction was quenched with sat. NH4Cl (aq.) and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (88/12, v/v) to afford 6-bromo-5-methoxy-[1,3]thiazolo[4,5-b]pyridin-2-amine (690.0 mg, 46%) as a white solid. LCMS (ESI, m/z): [M+H]+=259.9.
To a solution of 6-bromo-5-methoxy-[1,3]thiazolo[4,5-b]pyridin-2-amine (690.0 mg, 2.65 mmol) in THF (10.0 mL) was added DMSO (18.6 mg, 0.24 mmol) and t-BuONO (410.3 mg, 3.98 mmol) at room temperature under N2. The resulting mixture was stirred at 30° C. for 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (92/8, v/v) to afford 6-bromo-5-methoxy-[1,3]thiazolo[4,5-b]pyridine (120.0 mg, 18%) as a white solid. LCMS (ESI, m/z): [M+H]+=244.9.
To a solution of 6-bromo-5-methoxy-[1,3]thiazolo[4,5-b]pyridine (120.0 mg, 0.49 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (240.2 mg, 0.58 mmol), K2CO3 (203.0 mg, 1.46 mmol) and Pd(dppf)Cl2 (79.5 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (65/35, v/v) to afford 6-chloro-3-[5-methoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (130.0 mg, 32%) as a white solid. LCMS (ESI, m/z): [M+H]+=447.1.
To a solution of 6-chloro-3-[5-methoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (100.0 mg, 0.22 mmol) in t-BuOH (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (115.3 mg, 1.12 mmol), XPhos (21.3 mg, 0.04 mmol), K2CO3 (92.7 mg, 0.61 mmol) and Pd(OAc)2 (5.0 mg, 0.02 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (65/35, v/v) to afford (1S,2S)-2-fluoro-N-(3-[5-methoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (47.0 mg, 41%) as a white solid. LCMS (ESI, m/z): [M+H]+=514.2.
To a solution of (1S,2S)-2-fluoro-N-(3-[5-methoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (47.0 mg, 0.09 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (3.0 mL) was added NH3·H2O (3 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 47% B in 10 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-[5-methoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (5.9 mg, 16%) as a white solid. LCMS (ESI, m/z): [M+H]+=384.1. 1H NMR (400 MHz, DMSO-d6): δ 11.80 (s, 1H), 10.68 (s, 1H), 9.51 (s, 1H), 8.81 (s, 1H), 8.25 (d, J=8.8 Hz, 1H), 7.95 (d, J=8.4 Hz, 1H), 7.84 (d, J=1.6 Hz, 1H), 5.04-4.83 (m, 1H), 4.06 (s, 3H), 2.26-2.23 (m, 1H), 1.72-1.63 (m, 1H), 1.24-1.12 (m, 1H).
To a solution of 3-fluoro-5-methoxyaniline (2.0 g, 14.17 mmol) in DMF (20.0 mL) was added NIS (3.2 g, 14.17 mmol) at room temperature. The resulting mixture was stirred at room temperature for 0.5 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 3-fluoro-4-iodo-5-methoxyaniline (2.5 g, 66%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=268.0.
To a solution of 3-fluoro-4-iodo-5-methoxyaniline (2.5 g, 9.36 mmol) in acetone (50.0 mL) was added benzoyl isothiocyanate (1.5 g, 9.36 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 30 min. After the reaction was completed, the reaction mixture was cooled to room temperature and filtered. The solid was collected and dried to afford N-((3-fluoro-4-iodo-5-methoxyphenyl)carbamothioyl)benzamide (3.8 g, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=431.0.
To a solution of N-((3-fluoro-4-iodo-5-methoxyphenyl)carbamothioyl)benzamide (3.8 g, 8.83 mmol) in MeOH/H2O (30.0/30.0 mL) was added NaOH (0.4 g, 9.7 mmol) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 1-(3-fluoro-4-iodo-5-methoxyphenyl)thiourea (1.9 g, 65%) as a white solid. LCMS (ESI, m/z): [M+H]+=326.9.
To a solution of 1-(3-fluoro-4-iodo-5-methoxyphenyl)thiourea (1.9 g, 5.83 mmol) in CHCl3 (30.0 mL) was added dropwise Br2 (1.0 g, 6.41 mmol) at 0° C. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was quenched with aq. NaHSO3 and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 7-fluoro-6-iodo-5-methoxybenzo[d]thiazol-2-amine (1.3 g, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=324.9.
To a solution of 7-fluoro-6-iodo-5-methoxybenzo[d]thiazol-2-amine (1.3 g, 4.01 mmol) in THE (30.0 mL) was added t-BuONO (0.6 g, 6.02 mmol) and DMSO (25.1 mg, 0.32 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 7-fluoro-6-iodo-5-methoxybenzo[d]thiazole (1.2 g, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=309.9.
To a solution of 7-fluoro-6-iodo-5-methoxybenzo[d]thiazole (300.0 mg, 0.97 mmol) in 1,4-dioxane/H2O (16.0/4.0 mL) was added (6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)boronic acid (317.0 mg, 0.97 mmol), K2CO3 (402.4 mg, 2.91 mmol) and Pd(dppf)Cl2 (71.0 mg, 0.10 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-7-fluoro-5-methoxybenzo[d]thiazole (270.0 mg, 59%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=464.1.
To a solution of 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-7-fluoro-5-methoxybenzo[d]thiazole (210.0 mg, 0.45 mmol) in dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (233.3 mg, 2.27 mmol), K2CO3 (187.6 mg, 1.36 mmol), Brettphos (48.6 mg, 0.09 mmol) and BrettPhos Pd G3 (41.0 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the mixture was evaporated in vacuo. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)-2-fluoro-N-(3-(7-fluoro-5-methoxybenzo[d]thiazol-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (160.0 mg, 66%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=531.2.
To a solution of (1S,2S)-2-fluoro-N-(3-(7-fluoro-5-methoxybenzo[d]thiazol-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (140.0 mg, 0.26 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 23% to 53% in 10 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-(7-fluoro-5-methoxybenzo[d]thiazol-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (33.5 mg, 31%) as a white solid. LCMS (ESI, m/z): [M+H]+=401.2. 1H NMR (300 MHz, DMSO-d6): δ 11.80 (s, 1H), 10.68 (s, 1H), 9.45 (s, 1H), 7.90 (d, J=8.4 Hz, 1H), 7.77-7.70 (m, 2H), 7.58 (s, 1H), 5.05-4.82 (m, 1H), 3.90 (s, 3H), 2.28-2.19 (m, 1H), 1.70-1.62 (m, 1H), 1.22-1.11 (m, 1H).
To a solution of cyclopropylmethanol (2.1 g, 28.58 mmol) in DMF (50.0 mL) was added NaH (1.9 g, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h under N2. Then 3-bromo-2-chloropyridine (5.0 g, 25.98 mmol) was added to the mixture at 0° C. under N2. The resulting mixture was stirred at 70° C. for another 6 h. After the reaction was completed, the reaction mixture was quenched with H2O at 0° C. and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 3-bromo-2-(cyclopropylmethoxy)pyridine (3.0 g, 50%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=228.0.
To a solution of 3-bromo-2-(cyclopropylmethoxy)pyridine (500.0 mg, 2.19 mmol) in 1,4-dioxane/H2O (16.0/4.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (896.2 mg, 2.19 mmol), K2CO3 (908.9 mg, 6.58 mmol) and Pd(dppf)Cl2 (160.4 mg, 0.22 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 6-chloro-3-(2-(cyclopropylmethoxy)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (500.0 mg, 53%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=430.2.
To a solution of 6-chloro-3-(2-(cyclopropylmethoxy)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (400.0 mg, 0.93 mmol) in t-BuOH (20.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (479.5 mg, 4.65 mmol), K2CO3 (385.7 mg, 2.79 mmol), Pd(OAc)2 (20.9 mg, 0.09 mmol) and XPhos (88.7 mg, 0.19 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford (1S,2S)—N-(3-(2-(cyclopropylmethoxy)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (160.0 mg, 34%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=497.2.
To a solution of (1S,2S)—N-(3-(2-(cyclopropylmethoxy)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (150.0 mg, 0.18 mmol) in DMF (5.0 mL) was added ethylenediamine (90.8 mg, 1.51 mmol) and TBAF (236.9 mg, 0.91 mmol) at room temperature. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) and then purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 44% to 54% in 8 min; 254 nm) to afford N-(3-(2-(cyclopropylmethoxy)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide Enantiomer 1 (5.2 mg, 4%, retention time: 7.38 minutes) as a white solid and N-(3-(2-(cyclopropylmethoxy)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide Enantiomer 2 (10.2 mg, 9%, retention time, 7.79 minutes) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 148 and 149 in Table 1.
N-(3-(2-(cyclopropylmethoxy)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide Enantiomer 1: LCMS (ESI, m/z): [M+H]+=367.2. 1H NMR (300 MHz, DMSO-d6): δ 11.73 (s, 1H), 10.69 (s, 1H), 8.21 (d, J=8.7 Hz, 1H), 8.05-7.92 (m, 3H), 7.80 (s, 1H), 7.08-7.04 (m, 1H), 5.04-4.82 (m, 1H), 4.22 (d, J=7.2 Hz, 2H), 2.26-2.22 (m, 1H), 1.78-1.55 (m, 1H), 1.31-1.13 (m, 2H), 0.59-0.53 (m, 2H), 0.39-0.34 (m, 2H).
N-(3-(2-(cyclopropylmethoxy)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide Enantiomer 2: LCMS (ESI, m/z): [M+H]+=367.2. 1H NMR (300 MHz, DMSO-d6): δ 11.75 (s, 1H), 10.81 (s, 1H), 8.20 (d, J=8.7 Hz, 1H), 8.05-7.98 (m, 2H), 7.89 (d, J=8.7 Hz, 1H), 7.80 (d, J=2.4 Hz, 1H), 7.07-7.03 (m, 1H), 5.02-4.78 (m, 1H), 4.21 (d, J=7.2 Hz, 2H), 2.69-2.55 (m, 1H), 1.55-1.48 (m, 1H), 1.29-1.22 (m, 2H), 0.57-0.52 (m, 2H), 0.38-0.35 (m, 2H).
To a solution of 3-bromo-2-methoxyaniline (5.0 g, 24.75 mmol) in acetone (125.0 mL) was added benzoyl isothiocyanate (4.0 g, 24.76 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 0.5 h. After the reaction was completed. The resulting mixture was concentrated under reduced pressure. The residue were washed with ethyl ether and then filtered. The solid was collected and dried to afford 1-benzoyl-3-(3-bromo-2-methoxyphenyl)thiourea (7.8 g, crude) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=365.0.
To a solution of 1-benzoyl-3-(3-bromo-2-methoxyphenyl)thiourea (7.8 g, crude) in MeOH/H2O (130.0 mL/26.0 mL) was added NaOH (4.3 g, 106.76 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 1 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 3-bromo-2-methoxyphenylthiourea (5.0 g, crude) as a light pink solid. LCMS (ESI, m/z): [M+H]+=261.0
To a solution of 3-bromo-2-methoxyphenylthiourea (5.0 g, 19.15 mmol) in CHCl3 (300.0 mL) was added dropwise Br2 (3.7 g, 22.98 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 8 h. After the reaction was completed, the reaction mixture was quenched with NaHSO3 (aq.) at 0° C. The resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with acetonitrile/water (1/1, v/v) to afford 5-bromo-4-methoxy-1,3-benzothiazol-2-amine (4.0 g, 80%) as a light brown solid. LCMS (ESI, m/z): [M+H]+=258.9.
To a solution of 5-bromo-4-methoxy-1,3-benzothiazol-2-amine (4.0 g, 15.45 mmol) in THE (60.0 mL) was added tert-butyl nitrite (2.4 g, 23.16 mmol) and DMSO (96.5 mg, 1.24 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (7/1, v/v) to afford 5-bromo-4-methoxy-1,3-benzothiazole (1.6 g, 42%) as a brown solid. LCMS (ESI, m/z): [M+H]+=243.9.
To a solution of 5-bromo-4-methoxy-1,3-benzothiazole (124.1 mg, 0.51 mmol) in 1,4-dioxane/H2O (10.0 mL/2.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (250.0 mg, 0.61 mmol), K2CO3 (140.5 mg, 1.02 mmol) and Pd(dppf)Cl2 (37.2 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxy-1,3-benzothiazole (220.0 mg, 97%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=446.1.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxy-1,3-benzothiazole (190.0 mg, 0.43 mmol) in t-BuOH (20.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (219.6 mg, 2.13 mmol), XPhos (40.6 mg, 0.09 mmol), K2CO3 (176.6 mg, 1.28 mmol) and Pd(OAc)2 (9.6 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(4-methoxy-1,3-benzothiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (114.0 mg, 52%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=513.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(4-methoxy-1,3-benzothiazol-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (114.0 mg, 0.22 mmol) in CH2Cl2 (6.0 mL) was added TFA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (6.0 mL) was added NH3·H2O (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 38% B to 47% B in 8 min; 254/220 nm) to afford (1S,2S)-2-fluoro-N-[3-(4-methoxy-1,3-benzothiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (16.7 mg, 19%) as a white solid. LCMS (ESI, m/z): [M+H]+=383.2. 1H NMR (400 MHz, DMSO-d6): δ 11.69 (s, 1H), 10.65 (s, 1H), 9.37 (s, 1H), 8.08 (d, J=8.4 Hz, 1H), 7.91 (d, J=8.4 Hz, 2H), 7.71-7.67 (m, 2H), 5.01-4.84 (m, 1H), 4.05 (s, 3H), 2.28-2.19 (m, 1H), 1.70-1.62 (m, 1H), 1.23-1.17 (m, 1H).
To a solution of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-formylcyclopropanecarboxamide (260.0 mg, 0.53 mmol) in DCM (26.0 mL) were added tert-butyl piperazine-1-carboxylate (293.1 mg, 1.57 mmol) and NaBH3CN (98.9 mg, 1.57 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford tert-butyl 4-[[trans-2-[[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]cyclopropyl]methyl]piperazine-1-carboxylate (180.0 mg, 52%) as a white solid. LCMS (ESI, m/z): [M+H]+=666.4.
To a solution of tert-butyl 4-[[trans-2-[[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]cyclopropyl]methyl]piperazine-1-carboxylate (350.0 mg, 0.53 mmol) in CH2Cl2 (10.0 mL) was added TFA (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (20.0 mL) was added NH3·H2O (20.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 43% B in 9 min; 254 nm) to afford trans-N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-(piperazin-1-ylmethyl)cyclopropane-1-carboxamide (80.0 mg, 39%) as a white solid. LCMS (ESI, m/z): [M+H]+=436.2.
The racemic trans-N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-(piperazin-1-ylmethyl)cyclopropane-1-carboxamide (80.0 mg, 0.18 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IE, 2×25 cm, 5 um; Mobile Phase A: MTBE (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: MeOH—HPLC; Flow rate: 20 mL/min; Gradient: 15% B to 15% B in 13 min; 220/254 nm) to afford N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-(piperazin-1-ylmethyl)cyclopropane-1-carboxamide Enantiomer 1 (12.4 mg, 15%, Retention time 1: 7.557 min) as a white solid and N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-(piperazin-1-ylmethyl)cyclopropane-1-carboxamide Enantiomer 2 (12.5 mg, 15%, Retention time 2: 9.866 min) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 151 and 152 in Table 1.
N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-(piperazin-1-ylmethyl)cyclopropane-1-carboxamide Enantiomer 1: LCMS (ESI, m/z): [M+H]+=436.3. 1H NMR (400 MHz, DMSO-d6): δ 11.40 (s, 1H), 10.51 (s, 1H), 7.81 (d, J=8.8 Hz, 1H), 7.52 (d, J=8.4 Hz, 1H), 7.29-7.25 (m, 2H), 6.74 (d, J=8.4 Hz, 2H), 3.68 (s, 6H), 2.73-2.67 (m, 4H), 2.38-2.32 (m, 5H), 2.22-2.18 (m, 1H), 1.89-1.87 (m, 1H), 1.35-1.31 (m, 1H), 1.04-0.98 (m, 1H), 0.78-0.61 (m, 1H).
N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-(piperazin-1-ylmethyl)cyclopropane-1-carboxamide Enantiomer 2: LCMS (ESI, m/z): [M+H]+=436.3. 1H NMR (400 MHz, DMSO-d6) δ 11.40 (s, 1H), 10.51 (s, 1H), 7.81 (d, J=8.8 Hz, 1H), 7.52 (d, J=8.8 Hz, 1H), 7.29-7.25 (m, 2H), 6.74 (d, J=8.4 Hz, 2H), 3.68 (s, 6H), 2.73-2.67 (m, 4H), 2.38-2.32 (m, 5H), 2.22-2.18 (m, 1H), 1.89-1.87 (m, 1H), 1.35-1.31 (m, 1H), 1.04-0.98 (m, 1H), 0.78-0.61 (m, 1H).
To a solution of 5-bromo-4-cyclopropoxy-6-methoxypyrimidine (500.0 mg, 2.04 mmol) in dioxane/H2O (20.0 mL/4.0 mL) was added 6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-ylboronic acid (666.5 mg, 2.04 mmol), K2CO3 (845.9 mg, 6.12 mmol) and Pd(dppf)Cl2 (149.3 mg, 0.20 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-cyclopropoxy-6-methoxypyrimidine (260.0 mg, 29%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=447.2.
To a solution of 5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-cyclopropoxy-6-methoxypyrimidine (240.0 mg, 0.54 mmol) in 1,4-dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (276.8 mg, 2.68 mmol), BrettPhos (57.6 mg, 0.11 mmol), Cs2CO3 (524.8 mg, 1.61 mmol) and BrettPhos Pd G3 (48.67 mg, 0.054 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)—N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (150.0 mg, 54%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=514.2.
To a solution of (1S,2S)—N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (150.0 mg, 0.12 mmol) in CH2Cl2 (10.0 mL) was added TFA (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The reaction mixture was evaporated in vacuo. To the residue in CH3CN (10.0 mL) was added NH3·H2O (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 31% B to 41% B in 8 min; 254 nm) to afford (1S,2S)—N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (17.2 mg, 20%) as a white solid. LCMS (ESI, m/z): [M+H]+=384.2. 1H NMR (400 MHz, DMSO-d6): δ 11.65 (s, 1H), 10.63 (s, 1H), 8.49 (s, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.67 (d, J=8.4 Hz, 1H), 7.45 (d, J=2.8 Hz, 1H), 5.01-4.83 (m, 1H), 4.38-4.34 (m, 1H), 3.92 (s, 3H), 2.28-2.22 (m, 1H), 1.68-1.61 (m, 1H), 1.18-1.14 (m, 1H), 0.82-0.71 (m, 2H), 0.69-0.61 (m, 2H).
To a solution of 4,6-dichloropyridin-2-amine (5.0 g, 30.68 mmol) in NMP (30.0 mL) was added CH3ONa (8.3 g, 153.38 mmol). The resulting mixture was stirred at 120° C. for 16 h. After the reaction was completed, the reaction was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (70/30, v/v) to afford 4,6-dimethoxypyridin-2-amine (2.7 g, 57%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=155.1.
To a solution of 4,6-dimethoxypyridin-2-amine (1.0 g, 6.49 mmol) in acetone (10.0 mL) was added benzoyl isothiocyanate (1.2 g, 7.14 mmol). The mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction mixture was filtered. The solid was collected and dried to afford 3-benzoyl-1-(4,6-dimethoxypyridin-2-yl)thiourea (1.8 g, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=318.1.
To a solution of 3-benzoyl-1-(4,6-dimethoxypyridin-2-yl)thiourea (1.7 g, 5.36 mmol) in MeOH/H2O (15.0 mL/3.0 mL) was added NaOH (428.5 mg, 10.71 mmol). The mixture was stirred at room temperature for 2 h. After the reaction was completed, the mixture was filtered. The solid was washed with H2O and dried to afford 4,6-dimethoxypyridin-2-ylthiourea (1.1 g, crude) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=214.1.
To a solution of 4,6-dimethoxypyridin-2-ylthiourea (1.0 g, 4.69 mmol) in CHCl3 (10.0 mL) was added dropwise Br2 (1.5 g, 9.38 mmol) at 0° C. The mixture was stirred at 60° C. for 1 h. After the reaction was completed, the reaction mixture was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (90/10, v/v) to afford 6-bromo-5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-2-amine (1.2 g, 88%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=290.0.
To a solution of 6-bromo-5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-2-amine (1.1 g, 3.79 mmol) in THF (10.0 mL) was added 2-methyl-2-propylnitrite (469.2 mg, 4.55 mmol) and DMSO (50.0 uL). The mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (90/10, v/v) to afford 6-bromo-5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridine (320.0 mg, 30%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=274.9.
To a solution of 6-bromo-5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridine (260.0 mg, 0.95 mmol) in 1,4-dioxane/H2O (5.0 mL/1.0 mL) was added 6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-ylboronic acid (308.7 mg, 0.95 mmol), K3PO4 (601.8 mg, 2.84 mmol) and PdAMPHOS (66.9 mg, 0.10 mmol). The mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the reaction mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (75/25, v/v) to afford 6-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridine (220.0 mg, 48%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=477.1.
To a solution of 6-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridine (200.0 mg, 0.42 mmol) in 1,4-dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (216.1 mg, 2.10 mmol), Cs2CO3 (409.7 mg, 1.257 mmol), BrettPhos (45.0 mg, 0.08 mmol) and BrettPhos Pd G3 (38.0 mg, 0.04 mmol) under N2. The mixture was stirred with microwave at 120° C. for 1 h. After the reaction was completed, the reaction was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (90/10, v/v) to afford (1S,2S)—N-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (160.0 mg, 70%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=544.2.
To a solution of (1S,2S)—N-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (140.0 mg, 0.26 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL). The mixture was stirred at room temperature for 3 h. The mixture was concentrated under vacuum. To the above mixture were added ACN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for another 3 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 35% B in 9 min; 254 nm) to afford (1S,2S)—N-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropane-1-carboxamide (31.0 mg, 29%) as a white solid. LCMS (ESI, m/z): [M+H]+=414.1. 1H NMR (300 MHz, DMSO-d6): δ 11.68 (s, 1H), 10.65 (s, 1H), 9.56 (s, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.71 (d, J=8.7 Hz, 1H), 7.48 (d, J=2.4 Hz, 1H), 5.06-4.78 (m, 1H), 3.90 (s, 6H), 2.28-2.23 (m, 1H), 1.72-1.58 (m, 1H), 1.18-1.05 (m, 1H).
To a solution of trans-2-(methoxycarbonyl)cyclopropane-1-carboxylic acid (144.3 mg, 1.00 mmol) in DMF (20.0 mL) were added EDCI (287.9 mg, 1.50 mmol) and HOBT (202.9 mg, 1.50 mmol) at room temperature. The resulting mixture was stirred at room temperature for 20 min. Then DIEA (517.5 mg, 4.00 mmol) and 3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (400.0 mg, 1.00 mmol) were added to the mixture at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the mixture was purified by reverse phase flash chromatography with H2O/CH3CN (1/1, v/v) to afford trans-methyl 2-[[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]cyclopropane-1-carboxylate (280.0 mg, 53%) as a white solid. LCMS (ESI, m/z): [M+H]+=526.2.
To a solution of trans-methyl 2-[[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]cyclopropane-1-carboxylate (330.0 mg, 0.63 mmol) in THF (33.0 mL)/CH3OH (33.0 mL) was added NaBH4 (712.5 mg, 18.83 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (300.0 mg, 96%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=498.2.
To a solution of trans-N-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (300.0 mg, 0.60 mmol) in CH2Cl2 (30.0 mL) was added Dess-Martin (767.0 mg, 1.81 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (260.0 mg, crude) as a brown solid. LCMS (ESI, m/z): [M+H]+=496.2.
To a solution of trans-N-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (350.0 mg, crude) in CH2Cl2 (20.0 mL) was added 1-methylpiperazine (212.2 mg, 2.12 mmol) and NaBH3CN (133.1 mg, 2.12 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was quenched with CH3OH. The resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (86/14, v/v) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (180.0 mg, 43%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=580.3.
To a solution of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (180.0 mg, 0.31 mmol) in DMF (5.0 mL) was added ethylenediamine (93.3 mg, 1.55 mmol) and TBAF (243.5 mg, 0.93 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (75/25, v/v) and then separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IE, 2×25 cm, 5 um; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 10% to 10% in 46 min; 220/254 nm) to afford N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide Enantiomer 1 (10.3 mg, 7%, Retention time 1: 16.876 min) as a white solid and N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide Enantiomer 1 (12.3 mg, 8%, Retention time 2: 30.093 min) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 155 and 156 in Table 1.
N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide Enantiomer 1: LCMS (ESI, m/z): [M+H]+=450.3. 1H NMR (400 MHz, DMSO-d6): δ 11.42 (s, 1H), 10.53 (s, 1H), 7.82 (d, J=8.4 Hz, 1H), 7.53 (d, J=8.8 Hz, 1H), 7.29-7.25 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 3.69 (s, 6H), 2.49-2.26 (m, 8H), 2.14 (s, 3H), 1.97-1.89 (m, 1H), 1.33-1.23 (m, 1H), 1.04-1.02 (m, 1H), 0.73-0.65 (m, 1H).
N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide Enantiomer 2: LCMS (ESI, m/z): [M+H]+=450.2. 1H NMR (400 MHz, DMSO-d6): δ 11.42 (s, 1H), 10.53 (s, 1H), 7.82 (d, J=7.6 Hz, 1H), 7.53 (d, J=8.0 Hz, 1H), 7.29-7.25 (m, 2H), 6.75 (d, J=8.0 Hz, 2H), 3.69 (s, 6H), 2.49-2.24 (m, 8H), 2.14 (s, 3H), 1.97-1.89 (m, 1H), 1.33-1.23 (m, 1H), 1.04-1.02 (m, 1H), 0.73-0.65 (m, 1H).
To a solution of trans-N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (400.0 mg, 0.80 mmol) in CH2Cl2 (5.0 mL) was added Dess-Martin (1022.7 mg, 2.41 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (500.0 mg, crude) as a brown solid. LCMS (ESI, m/z): [M+H]+=496.2.
To a solution of trans-N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (500.0 mg, crude) in CH2Cl2 (10.0 mL) was added morpholine (262.6 mg, 3.03 mmol) and NaBH3CN (189.4 mg, 3.03 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum CH2Cl2/MeOH (95/5, v/v) to afford trans-N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(morpholin-4-ylmethyl)cyclopropane-1-carboxamide (410.0 mg, 59%) as a brown solid. LCMS (ESI, m/z): [M+H]+=567.3.
To a solution of trans-N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(morpholin-4-ylmethyl)cyclopropane-1-carboxamide (390.0 mg, 0.69 mmol) in DMF (5.0 mL) was added TBAF (539.7 mg, 2.06 mmol) and DEA (206.8 mg, 3.44 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum CH2Cl2/MeOH (90/10, v/v) and then purified by reverse phase flash chromatography with ACN/H2O (50/50, v/v) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(morpholinomethyl)cyclopropanecarboxamide (100.0 mg, 30%) as a white solid. LCMS (ESI, m/z): [M+H]+=437.2.
The racemic trans-N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(morpholinomethyl)cyclopropanecarboxamide (100.0 mg, 0.23 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IE, 2×25 cm, 5 um; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 18 mL/min; Gradient: 50% B to 50% B in 23 min; 254/220 nm) to afford N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-(morpholin-4-ylmethyl)cyclopropane-1-carboxamide Enantiomer 1 (22.2 mg, 44%, Retention time 1: 11.1 min) as a white solid and (1S,2S)—N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-(morpholin-4-ylmethyl)cyclopropane-1-carboxamide Enantiomer 2 (assumed, 28.4 mg, 56%, Retention time 2: 15.904 min) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 157 and 158 in Table 1.
N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-(morpholin-4-ylmethyl)cyclopropane-1-carboxamide Enantiomer 1: LCMS (ESI, m/z): [M+H]+=437.3. 1H NMR (400 MHz, DMSO-d6): δ 11.42 (s, 1H), 10.54 (s, 1H), 7.81 (d, J=8.4 Hz, 1H), 7.52 (d, J=8.4 Hz, 1H), 7.30-7.25 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 3.69 (s, 6H), 3.59-3.57 (m, 4H), 2.50-2.43 (m, 4H), 2.38-2.33 (m, 1H), 2.27-2.25 (m, 1H), 1.95-1.86 (m, 1H), 1.41-1.32 (m, 1H), 1.11-0.99 (m, 1H), 0.77-0.65 (m, 1H).
N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-(morpholin-4-ylmethyl)cyclopropane-1-carboxamide Enantiomer 2: LCMS (ESI, m/z): [M+H]+=437.3. 1H NMR (400 MHz, DMSO-d6) δ 11.42 (s, 1H), 10.54 (s, 1H), 7.81 (d, J=8.8 Hz, 1H), 7.52 (d, J=8.8 Hz, 1H), 7.30-7.25 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 3.69 (s, 6H), 3.59-3.57 (m, 4H), 2.50-2.43 (m, 4H), 2.38-2.33 (m, 1H), 2.27-2.25 (m, 1H), 1.95-1.86 (m, 1H), 1.41-1.32 (m, 1H), 1.11-0.99 (m, 1H), 0.77-0.65 (m, 1H).
The solution of 2,5-dibromo-4-methoxypyridine (500.0 mg, 2.24 mmol) in NH2NH2·H2O (50.0 mL, 80%) was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with CH3CN/H2O (1/4, v/v) to afford 5-bromo-2-hydrazinyl-4-methoxypyridine (60.0 mg, 12%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=218.0
The solution of 5-bromo-2-hydrazinyl-4-methoxypyridine (600.0 mg, 2.75 mmol) in formic acid (10.0 mL) was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with MeOH/H2O (1/10, v/v) to afford 6-bromo-7-methoxy-[1,2,4]triazolo[4,3-a]pyridine (120.0 mg, 19%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=228.0
To a mixture of 6-bromo-7-methoxy-[1,2,4]triazolo[4,3-a]pyridine (100.0 mg, 0.43 mmol) and 6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-ylboronic acid (214.8 mg, 0.65 mmol) in dioxane/H2O (5.0/1.0 mL) were added Pd(dppf)Cl2 (32.0 mg, 0.04 mmol) and K2CO3 (181.8 mg, 1.31 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was residue was purified by flash column chromatography with MeOH/DCM (1/10, v/v) to afford 6-chloro-3-[7-methoxy-[1,2,4]triazolo[4,3-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (110.0 mg, 58%) as a white solid. LCMS (ESI, m/z): [M+H]+=430.1
To a mixture of 6-chloro-3-[7-methoxy-[1,2,4]triazolo[4,3-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (90.0 mg, 0.20 mmol) and (1S,2S)-2-fluorocyclopropane-1-carboxamide (107.9 mg, 1.04 mmol) in dioxane (4.0 mL) was added BrettPhos Pd G3 (37.5 mg, 0.04 mmol), BrettPhos (44.9 mg, 0.08 mmol) and Cs2CO3 (204.0 mg, 0.62 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with MeOH/DCM (1/10, v/v) to afford (1S,2S)-2-fluoro-N-(3-[7-methoxy-[1,2,4]triazolo[4,3-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (40.0 mg, 38%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=497.2
To a solution of (1S,2S)-2-fluoro-N-(3-[7-methoxy-[1,2,4]triazolo[4,3-a]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (30.0 mg, 0.06 mmol) in DCM (1.0 mL) was added TFA (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under vacuum. To the above residue was added ACN (2.0 mL) and NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 16% B to 26% B in 8 min, 254 nm) to afford (1S,2S)-2-fluoro-N-(3-[7-methoxy-[1,2,4]triazolo[4,3-a]pyridin-6-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (Compound 159) (5.0 mg, 22%) as a white solid. LCMS (ESI, m/z): [M+H]+=367.1. 1H NMR (300 MHz, DMSO-d6): δ 11.77 (s, 1H), 10.68 (s, 1H), 9.06 (s, 1H), 8.68 (s, 1H), 8.12 (d, J=8.7 Hz, 1H), 7.94 (d, J=8.4 Hz, 1H), 7.68 (s, 1H), 7.20 (s, 1H), 5.07-4.80 (m, 1H), 3.95 (s, 3H), 2.27-2.24 (m, 1H), 1.73-1.60 (m, 1H), 1.22-1.10 (m, 1H).
To a mixture of 5-bromo-6-methoxypyridin-3-amine (1.0 g, 4.92 mmol) and KSCN (238.8 mg, 2.46 mmol) in AcOH (20.0 mL) was added Br2 (390.0 mg, 2.46 mmol) at 0° C. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the pH value of the mixture was adjusted to 9 with aq·NaHCO3. The resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 6-bromo-5-methoxy-[1,3]thiazolo[5,4-b]pyridin-2-amine (2.0 g, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=259.9
To a mixture of 6-bromo-5-methoxy-[1,3]thiazolo[5,4-b]pyridin-2-amine (1.3 g, 4.99 mmol) and DMSO (312.4 mg, 3.99 mmol) in THE (20.0 mL) was added dropwise 2-methyl-2-propylnitrite (1.5 g, 15.00 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 6-bromo-5-methoxy-[1,3]thiazolo[5,4-b]pyridine (800.0 mg, 65%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=244.9
To a mixture of 6-bromo-5-methoxy-[1,3]thiazolo[5,4-b]pyridine (400.0 mg, 1.63 mmol) and 6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-ylboronic acid (1.0 g, 2.44 mmol) in dioxane/H2O (10.0/1.0 mL) was added K2CO3 (676.6 mg, 4.89 mmol) and Pd(dppf)Cl2 (119.4 mg, 0.16 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (9/1, v/v) to afford 6-chloro-3-[5-methoxy-[1,3]thiazolo[5,4-b]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (370.0 mg, 50%) as a purple solid. LCMS (ESI, m/z): [M+H]+=447.1
To a mixture of 6-chloro-3-[5-methoxy-[1,3]thiazolo[5,4-b]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy] methyl]pyrrolo[2,3-b]pyridine (350.0 mg, 0.78 mmol) and (1S,2S)-2-fluorocyclopropane-1-carboxamide (403.5 mg, 3.91 mmol) in BuOH (5.0 mL) was added Pd(OAc)2 (35.1 mg, 0.15 mmol), K2CO3 (324.6 mg, 2.34 mmol) and X-Phos (37.3 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)-2-fluoro-N-(3-[5-methoxy-[1,3]thiazolo[5,4-b]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (50.0 mg, 12%) as a green solid. LCMS (ESI, m/z): [M+H]+=514.2
To a solution of (1S,2S)-2-fluoro-N-(3-[5-methoxy-[1,3]thiazolo[5,4-b]pyridin-6-yl]-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (40.0 mg, 0.07 mmol) in DCM (1.0 mL) was added TFA (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added ACN (1.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 37% B to 46% B in 8 min, 254 nm) to afford (1S,2S)-2-fluoro-N-(3-[5-methoxy-[1,3]thiazolo[5,4-b]pyridin-6-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (Compound 160) (9.2 mg, 31%) LCMS (ESI, m/z): [M+H]+=384.1. 1H NMR (300 MHz, DMSO-d6): δ 11.81 (s, 1H), 10.68 (s, 1H), 9.33 (s, 1H), 8.54 (s, 1H), 8.13 (d, J=8.7 Hz, 1H), 7.95 (d, J=8.7 Hz, 1H), 7.80 (d, J=1.5 Hz, 1H), 5.10-4.80 (m, 1H), 4.05 (s, 3H), 2.25-2.15 (m, 1H), 1.78-1.60 (m, 1H), 1.27-1.10 (m, 1H).
To a solution of 3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (180.0 mg, 0.42 mmol) in DCM (13.0 mL) was added pyridine (133.5 mg, 1.69 mmol) and phenyl chloroformate ((132.1 mg, 0.84 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in pyridine (13.0 mL) was added (2-aminoethyl)dimethylamine (372.0 mg, 4.22 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (9/1, v/v) to afford 1-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[2-(dimethylamino)ethyl]urea (180.0 mg, 79%) as a white solid. LCMS (ESI, m/z): [M+H]+=541.3.
To a solution of 1-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[2-(dimethylamino)ethyl]urea (180.0 mg, 0.33 mmol) in CH2Cl2 (8.0 mL) was added TFA (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (8.0 mL) was added NH3·H2O (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column (XBridge Prep OBD C18 Column, 30×150 mm, 5 m; Mobile Phase A: Water (10 mmol/L NH4HCO3). Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 20% B to 36% B in 8 min; 254 nm) to afford 1-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(2-(dimethylamino)ethyl)urea (Compound 161) (49.6 mg, 36%) as a white solid. LCMS (ESI, m/z): [M+H]+=411.4. 1H NMR (400 MHz, DMSO-d6): δ 11.40 (s, 1H), 9.16 (s, 1H), 8.43 (s, 1H), 8.06 (d, J=6.0 Hz, 1H), 7.49 (d, J=8.8 Hz, 1H), 7.23 (d, J=2.0 Hz, 1H), 7.13 (d, J=6.0 Hz, 1H), 7.00 (d, J=8.4 Hz, 1H), 3.94-3.90 (m, 1H), 3.81 (s, 3H), 3.34-3.28 (m, 2H), 2.42-2.34 (m, 2H), 2.21 (s, 6H), 0.81-0.76 (m, 2H), 0.65-0.61 (m, 2H).
To a solution of 5-fluoro-2-methoxypyridin-4-ol (400.0 mg, 2.79 mmol) in MeCN (5.0 mL) was added NIS (628.8 mg, 2.79 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 5-fluoro-3-iodo-2-methoxypyridin-4-ol (450.0 mg, crude) as yellow solid. LCMS (ESI, m/z): [M+H]+=269.9
To a mixture of 5-fluoro-3-iodo-2-methoxypyridin-4-ol (700.0 mg, crude) and Ag2CO3 (2.8 g, 10.40 mmol) in CHCl3 (15.0 mL) was added CH3CH2I (1.5 g, 10.40 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 4-ethoxy-5-fluoro-3-iodo-2-methoxypyridine (700.0 mg, 90%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=298.0.
To a mixture of 4-ethoxy-5-fluoro-3-iodo-2-methoxypyridine (200.0 mg, 0.67 mmol) and 6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-ylboronic acid (263.9 mg, 0.80 mmol) in dioxane/H2O (5.0/0.5 mL) was added Pd(PPh3)4 (155.6 mg, 0.13 mmol) and K2CO3 (279.1 mg, 2.02 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-ethoxy-5-fluoro-2-methoxypyridine (140.0 mg, 46%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=452.1
To a mixture of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-ethoxy-5-fluoro-2-methoxypyridine (100.0 mg, 0.22 mmol) and (1S,2S)-2-fluorocyclopropane-1-carboxamide (114.0 mg, 1.10 mmol) in t-BuOH (4.0 mL) was added X-Phos (42.1 mg, 0.08 mmol), Pd(OAc)2 (9.93 mg, 0.044 mmol) and K2CO3 (91.7 mg, 0.66 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)—N-[3-(4-ethoxy-5-fluoro-2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (80.0 mg, 69%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=519.2
To a solution of (1S,2S)—N-[3-(4-ethoxy-5-fluoro-2-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (70.0 mg, 0.13 mmol) in DCM (2.0 mL) was added TFA (2.0 mL). The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under vacuum. To the above residue was added ACN (2.0 mL) and NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um, Mobile Phase A: Water (10 mmol/L NH4HCO3, Mobile Phase B ACN; Flow rate: 60 mL/min; Gradient: 39% B to 49% B in 9 min, 254/220 nm) to afford (1S,2S)—N-[3-(4-ethoxy-5-fluoro-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (Compound 162) (5.4 mg, 10%) as a white solid. LCMS (ESI, m/z): [M+H]+=389.2. 1H NMR (300 MHz, DMSO-d6): δ 11.71 (s, 1H), 10.65 (s, 1H), 8.11 (d, J=2.7 Hz, 1H), 7.87 (d, J=8.4 Hz, 1H), 7.73 (d, J=8.7 Hz, 1H), 7.49 (d, J=2.7 Hz, 1H), 5.05-4.81 (m, 1H), 4.07-4.00 (m, 2H), 3.35 (s, 3H), 2.31-2.19 (m, 1H), 1.72-1.62 (m, 1H), 1.22-1.18 (m, 1H), 1.11-1.02 (m, 3H).
To a solution of 3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (150.0 mg, 0.33 mmol) in CH2C2 (2.0 mL) was added phenyl carbonochloridate (77.0 mg, 0.49 mmol) and pyridine (1.0 mL) at 0° C. The resulting mixture was stirred at 0° C. for 1 h. Then a solution of (2-aminoethyl)dimethylamine (260.6 mg, 2.95 mmol) in pyridine (3.0 mL) was added dropwise to the mixture at 0° C. The resulting mixture was stirred at 60° C. for another 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (50/50, v/v) to afford 1-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(2-(dimethylamino)ethyl)urea (100.0 mg, 86%) as a white solid. LCMS (ESI, m/z): [M+H]+=572.2.
To a solution of 1-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(2-(dimethylamino)ethyl)urea (100.0 mg, 0.21 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L FA), Mobile Phase B: MeOH; Flow rate: 25 mL/min; Gradient: 26% B to 26% B in 15 min; 254 nm) to afford 1-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(2-(dimethylamino)ethyl)urea diformate (Compound 163) (16.5 mg, 14%) as a white solid. LCMS (ESI, m/z): [M+H]+=442.3. 1H NMR (400 MHz, DMSO-d6): δ 11.56 (s, 1H), 9.56 (s, 1H), 9.19 (s, 1H), 8.43 (s, 1H), 8.24 (s, 2H), 7.63 (d, J=8.8 Hz, 1H), 7.36 (s, 1H), 7.06 (d, J=8.4 Hz, 1H), 3.92-3.89 (m, 6H), 3.39-3.33 (m, 2H), 2.51-2.45 (m, 2H), 2.25 (s, 6H).
To a solution of methyl 2-amino-3-hydroxypropanoate hydrochloride (10.0 g, 64.28 mmol) in DMF (600.0 mL) was added KI (5.3 g, 32.14 mmol), K2CO3 (26.7 g, 192.83 mmol) and benzyl bromide (23.1 g, 134.98 mmol) dropwise at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (11/4, v/v) to afford methyl 2-(dibenzylamino)-3-hydroxypropanoate (15.1 g, 78%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=300.2.
To a solution of methyl 2-(dibenzylamino)-3-hydroxypropanoate (7.0 g, 23.38 mmol) in DCM (300.0 mL) was added dropwise DAST (9.4 g, 58.46 mmol) at −40° C. under N2. The reaction mixture was stirred at room temperature for 16 h. After the reaction was completed. The reaction mixture was quenched with sat. NH4Cl (aq.) at 0° C. The resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (9/2, v/v) to afford methyl 3-(dibenzylamino)-2-fluoropropanoate (4.9 g, 68%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=302.1.
To a solution of LiAlH4 (1.9 g, 49.77 mmol) in THE (200.0 mL) was added dropwise a solution of methyl 3-(dibenzylamino)-2-fluoropropanoate (5 g, 16.59 mmol) in THE at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h under N2. After the reaction was completed. The reaction was quenched with water at 0° C. The pH value of the mixture was adjusted to 5 with HCl (aq.). The resulting mixture was extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (7/3, v/v) to afford 3-(dibenzylamino)-2-fluoropropan-1-ol (3.4 g, 74%) as a white solid. LCMS (ESI, m/z): [M+H]+=274.2.
To a solution of 3-(dibenzylamino)-2-fluoropropan-1-ol (5.2 g, 19.02 mmol) in CH2Cl2 (300.0 mL) was added TEA (2.9 g, 28.54 mmol) and methanesulfonyl chloride (2.4 g, 20.93 mmol) at 0° C. The resulting mixture was stirred at room temperature for 6 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 3-(dibenzylamino)-2-fluoropropyl methanesulfonate (6.6 g, crude) as a yellow oil. LCMS (ESI, m/z): [M+H]+=352.1.
The mixture of 3-(dibenzylamino)-2-fluoropropyl methanesulfonate (4.0 g, crude) in dimethylamine/water (120.0 mL, 30%) was stirred at 60° C. for 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford [3-(dibenzylamino)-2-fluoropropyl]dimethylamine (3.3 g, crude) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=301.2.
To a solution of [3-(dibenzylamino)-2-fluoropropyl]dimethylamine (3.8 g, 12.65 mmol) in MeOH (200.0 mL) was added Pd(OH)2/C (1.1 g, wet) at room temperature. The resulting mixture was stirred at 60° C. for 16 h under H2. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure to afford (3-amino-2-fluoropropyl)dimethylamine (1.4 g, crude) as a colorless oil. LCMS (ESI, m/z): [M+H]+=121.1.
To a solution of 3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (300.0 mg, 0.75 mmol) in DCM (12.0 mL) was added pyridine (237.6 mg, 3.00 mmol) and phenyl chloroformate (235.1 mg, 1.50 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added (3-amino-2-fluoropropyl)dimethylamine (902.3 mg, 7.51 mmol) and pyridine (20.0 mL) at room temperature. The resulting mixture was stirred at 60° C. for 16 h. After the reaction was completed. the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (13/1, v/v) to afford 1-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea (336.0 mg, 82%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=546.3.
To a solution of 1-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea (300.0 mg, 0.55 mmol) in CH2Cl2 (10.0 mL) was added TFA (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (10.0 mL) was added NH3·H2O (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with acetonitrile/water (7/4, v/v) to afford 1-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea (106.0 mg, 46%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=416.2.
The racemic 1-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea (90.0 mg, 0.22 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IA, 2×25 cm, 20 um; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 50% B to 50% B in 14 min; 220/254 nm) to afford 1-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 1 (12.6 mg, 14%) as a white solid and 1-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 2 (16.9 mg, 18%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 164 and 165 in Table 1.
1-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 1: Retention Time 1: 7.811 min; LCMS (ESI, m/z): [M+H]+=416.1. 1H NMR (300 MHz, CD3OD): δ 7.59 (d, J=8.7 Hz, 1H), 7.31-7.26 (m, 1H), 7.21 (s, 1H), 6.75 (d, J=8.4 Hz, 2H), 6.68 (d, J=8.4 Hz, 1H), 5.00-4.80 (m, 1H), 3.77-3.71 (m, 7H), 3.69-3.50 (m, 1H), 2.78-2.71 (m, 1H), 2.69-2.50 (m, 1H), 2.36 (s, 6H).
1-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 2: Retention Time 2: 11.45 min; LCMS (ESI, m/z): [M+H]+=416.1. 1H NMR (300 MHz, CD3OD): δ 7.59 (d, J=8.4 Hz, 1H), 7.31-7.26 (m, 1H), 7.21 (s, 1H), 6.75 (d, J=8.4 Hz, 2H), 6.68 (d, J=8.4 Hz, 1H), 5.00-4.80 (m, 1H), 3.77-3.71 (m, 6H), 3.69-3.50 (m, 2H), 2.79-2.71 (m, 1H), 2.69-2.50 (m, 1H), 2.36 (s, 6H).
The solution of 2,3-difluoro-1-methoxy-4-nitrobenzene (2.0 g, 10.57 mmol) in NH3/MeOH (20.0 mL, 7 mol/L) was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 2-fluoro-3-methoxy-6-nitroaniline (1.8 g, 91%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=187.0
The mixture of 2-fluoro-3-methoxy-6-nitroaniline (1.5 g, 8.05 mmol) and NBS (2.1 g, 12.08 mmol) in AcOH (20.0 mL) was stirred at 90° C. for 4 h. After the reaction was completed, the reaction mixture was diluted with water and then filtered. The solid was washed with H2O and then collected to afford 4-bromo-2-fluoro-3-methoxy-6-nitroaniline (2.5 g, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=265.0
To a mixture of 4-bromo-2-fluoro-3-methoxy-6-nitroaniline (1.5 g, 5.66 mmol) and Fe (3.1 g, 56.60 mmol) in i-PrOH (10.0 mL) was added NH4Cl (2.1 g, 39.62 mmol) and formic acid (10.0 mL) at room temperature. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (4/1, v/v) to afford 6-bromo-4-fluoro-5-methoxy-1H-1,3-benzodiazole (620.0 mg, 44%) as a brown solid. LCMS (ESI, m/z): [M+H]+=245.0.
To a solution of 6-bromo-4-fluoro-5-methoxy-1H-1,3-benzodiazole (500.0 mg, 2.04 mmol) in THE (5.0 mL) was added NaH (146.8 mg, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 2 h. Then [2-(chloromethoxy)ethyl]trimethylsilane (510.2 mg, 3.06 mmol) was added to the mixture at 0° C. The resulting mixture was stirred at room temperature for additional 2 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (4/1, v/v) to afford 6-bromo-4-fluoro-5-methoxy-1-{[2-(trimethylsilyl)ethoxy]methyl}-1,3-benzodiazole (700.0 mg, 91%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=375.0
To a mixture of 6-bromo-4-fluoro-5-methoxy-1-{[2-(trimethylsilyl)ethoxy]methyl}-1,3-benzodiazole (350.0 mg, 0.93 mmol) and 6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-ylboronic acid (609.2 mg, 1.86 mmol) in 1,4-dioxane/H2O (5.0/0.5 mL) were added Pd(dppf)Cl2 (136.4 mg, 0.18 mmol) and K2CO3 (386.6 mg, 2.79 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) afford 6-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-4-fluoro-5-methoxy-1-{[2-(trimethylsilyl)ethoxy]methyl}-1,3-benzodiazole (200.0 mg, 37%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=577.2.
To a mixture of 6-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-4-fluoro-5-methoxy-1-{[2-(trimethylsilyl)ethoxy]methyl}-1,3-benzodiazole (190.0 mg, 0.32 mmol) and (1S,2S)-2-fluorocyclopropane-1-carboxamide (169.6 mg, 1.64 mmol) in 1,4-dioxane (4.0 mL) were added Pd2(dba)3 (60.2 mg, 0.06 mmol), BrettPhos (70.7 mg, 0.13 mmol) and Cs2CO3 (321.7 mg, 0.98 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/2, v/v) to afford (1S,2S)-2-fluoro-N-[3-(7-fluoro-6-methoxy-3-{[2-(trimethylsilyl)ethoxy]methyl}-1,3-benzodiazol-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (100.0 mg, 47%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=644.3.
To a solution of (1S,2S)-2-fluoro-N-[3-(7-fluoro-6-methoxy-3-{[2-(trimethylsilyl)ethoxy]methyl}-1,3-benzodiazol-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (100.0 mg, 0.15 mmol) in DCM (1.0 mL) was added TFA (1.0 mL). The mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added NH3·H2O (2.0 mL) and ACN (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 14% B to 34% B in 9 min, 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(7-fluoro-6-methoxy-3H-1,3-benzodiazol-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (Compound 166) (5.2 mg, 9%) as a white solid. LCMS (ESI, m/z): [M+H]+=384.1. 1H NMR (400 MHz, DMSO-d6): δ 12.73-12.68 (br, 1H), 11.70 (s, 1H), 10.69 (s, 1H), 8.25 (s, 1H), 8.06 (d, J=8.4 Hz, 1H), 7.94 (d, J=8.0 Hz, 1H), 7.65 (s, 1H), 7.50 (s, 1H), 5.02-4.85 (m, 1H), 3.62 (s, 3H), 2.32-2.25 (m, 1H), 1.70-1.64 (m, 1H), 1.23-1.16 (m, 1H).
To a solution of 3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (200.0 mg, 0.47 mmol) in DCM (4.0 mL) was added phenyl chloroformate (87.9 mg, 0.56 mmol) and pyridine (148.0 mg, 1.87 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure. To the residue in pyridine (4.0 mL) was added [2-(2-aminoethoxy)ethyl]dimethylamine (185.5 mg, 1.40 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with CH3CN/H2O (80/20, v/v) to afford 1-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-{2-[2-(dimethylamino)ethoxy]ethyl}urea (100.0 mg, 36%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=586.3.
To a solution of 1-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-{2-[2-(dimethylamino)ethoxy]ethyl}urea (100 mg, 0.17 mmol) in DCM (1.0 mL) was added TFA (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in ACN (1.0 mL) was added NH3·H2O (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 5% B to 30% B in 7 min, 254 nm) to afford 1-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-{2-[2-(dimethylamino)ethoxy]ethyl}urea; formic acid (Compound 167) (3.9 mg, 4%) as a white solid. LCMS (ESI, m/z): [M+H]+=456.2. 1H NMR (400 MHz, DMSO-d6): δ 11.74 (s, 1H), 9.25 (s, 1H), 8.70 (d, J=4.0 Hz, 1H), 8.50 (s, 1H), 8.22-8.19 (m, 1H) 7.58 (d, J=8.8 Hz, 1H), 7.35 (d, J=2.0 Hz, 1H), 6.99 (d, J=8.0 Hz, 1H), 4.37-4.34 (m, 1H), 3.91 (s, 3H), 3.63-3.59 (m, 2H), 3.54-3.45 (m, 2H), 3.39-3.34 (m, 2H), 2.68-2.60 (m, 1H), 2.34-2.14 (m, 7H), 0.77-0.69 (m, 2H), 0.65-0.61 (m, 2H).
To a solution of 3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (190.0 mg, 0.44 mmol) in CH2Cl2 (4.0 mL) was added pyridine (140.6 mg, 1.78 mmol) and phenyl chloroformate (83.5 mg, 0.53 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure. To the residue in pyridine (4.0 mL) was added (3-amino-2-fluoropropyl)dimethylamine (534.0 mg, 4.44 mmol) at room temperature. The resulting mixture was stirred at 60° C. for another 1 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum CH2Cl2/CH3OH (95/5, v/v) to afford 1-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea (160.0 mg, 62%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=574.3.
To a solution of 1-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea (150.0 mg, 0.26 mmol) in CH2Cl2 (4.0 mL) was added TFA (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (4.0 mL) was added NH3·H2O (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was Prep-Chiral-HPLC with the following conditions (Column: CHIRAL ART Amylose-SA, 2×25 cm, 5 um; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 50% B to 50% B in 10.5 min; 254 nm) to afford 1-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 1 (10.2 mg, 8%) as a white solid and 1-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 2 (11.0 mg, 9%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 168 and 169 in Table 1.
1-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 1: Retention Time 1: 5.876 min; LCMS (ESI, m/z): [M+H]+=444.1. 1H NMR (400 MHz, DMSO-d6): δ 11.59 (s, 1H), 9.25 (s, 1H), 8.58 (s, 1H), 8.49 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.33 (s, 1H), 7.04 (d, J=8.4 Hz, 1H), 4.81-4.70 (m, 1H), 4.37-4.33 (m, 1H), 3.92 (s, 3H), 3.64-3.52 (m, 1H), 3.45-3.38 (m, 1H), 2.22 (s, 6H), 0.86-0.82 (m, 2H), 0.69-0.62 (m, 2H).
1-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 2: Retention Time 2: 9.106 min; LCMS (ESI, m/z): [M+H]+=444.2. 1H NMR (400 MHz, DMSO-d6): δ 11.60 (s, 1H), 9.27 (s, 1H), 8.60 (s, 1H), 8.49 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.33 (d, J=2.4 Hz, 1H), 7.03 (d, J=8.8 Hz, 1H), 4.81-4.68 (m, 1H), 4.37-4.33 (m, 1H), 3.91 (s, 3H), 3.64-3.52 (m, 1H), 3.43-3.34 (m, 1H), 2.21 (s, 6H), 0.85-0.72 (m, 2H), 0.72-0.62 (m, 2H).
To a solution of 3-buten-1-ol (10.0 g, 138.68 mmol) in tetrahydrofuran (100.0 mL) was added 3,4-dihydro-2H-pyran (2.4 g, 13.88 mmol) and 4-methylbenzenesulfonic acid (17.5 g, 208.01 mmol) at 0° C. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 2-(but-3-en-1-yloxy)tetrahydro-2H-pyran (9.3 g, 43%) as a colorless oil.
To a mixture of 2-(but-3-en-1-yloxy)tetrahydro-2H-pyran (800.0 mg, 5.11 mmol) and Rh2 (OAc)4 (22.63 mg, 0.05 mmol) in CH2Cl2 (5.0 mL) was added dropwise ethyl diazoacetate (1.1 g, 10.22 mmol) at room temperature during 8 h under N2. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford trans-ethyl 2-[2-(oxan-2-yloxy)ethyl]cyclopropane-1-carboxylate (300.0 mg, 24%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=243.2.
To a solution of trans-ethyl 2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)cyclopropane-1-carboxylate (300.0 mg, 0.82 mmol) in THF (5.0 mL) was added 3-(2-methoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (590.15 mg, 2.43 mmol) and AlMe3 (0.2 mL, 2 mol/L) at 0° C. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by reverse flash column chromatography with MeOH/H2O (5/80, v/v) to afford trans-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-[2-(oxan-2-yloxy)ethyl]cyclopropane-1-carboxamide (100.0 mg, 33%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=566.3.
To a solution of trans-N-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-[2-(oxan-2-yloxy)ethyl]cyclopropane-1-carboxamide (100.0 mg, 0.17 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 36% B to 46% B in 8 min; 254 nm) to afford trans-2-(2-hydroxyethyl)-N-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (Compound 170) (8.7 mg, 14%) as a white solid. LCMS (ESI, m/z): [M+H]+=352.1. 1H NMR (300 MHz, DMSO-d6): δ 11.55 (d, J=2.4 Hz, 1H), 10.50 (s, 1H), 7.99-7.89 (m, 2H), 7.57-7.53 (m, 2H), 7.29-7.24 (m, 1H), 7.11-7.00 (m, 2H), 4.55-4.52 (m, 1H), 3.81 (s, 3H), 3.54-3.48 (m, 2H), 1.86-1.84 (m, 1H), 1.48-1.42 (m, 2H), 1.34-1.28 (m, 1H), 1.04-1.01 (m, 1H), 0.71-0.68 (m, 1H).
To a solution of 3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (500.0 mg, 1.25 mmol) in DMF (10.0 mL) was added cis-2-(methoxycarbonyl)cyclopropane-1-carboxylic acid (180.4 mg, 1.25 mmol), DIEA (808.6 mg, 6.26 mmol) and HATU (570.9 mg, 1.50 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 2 h under N2. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (62/38, v/v) to afford methyl cis-2-((3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)carbamoyl)cyclopropane-1-carboxylate (640.0 mg, 97%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=526.2.
To a solution of methyl cis-2-((3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)carbamoyl)cyclopropane-1-carboxylate (640.0 mg, 1.21 mmol) in THF/CH3OH (10.0 mL/10.0 mL) was added NaBH4 (460.6 mg, 12.17 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was quenched with H2O at 0° C. and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford cis-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(hydroxymethyl)cyclopropane-1-carboxamide (510.0 mg, crude) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=498.2.
To a solution of cis-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(hydroxymethyl)cyclopropane-1-carboxamide (510.0 mg, crude) in CH2Cl2 (20.0 mL) was added Dess-Martin (869.3 mg, 2.06 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford cis-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-formylcyclopropane-1-carboxamide (840.0 mg, crude) as a brown solid. LCMS (ESI, m/z): [M+H]+=496.2.
To a solution of cis-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-formylcyclopropane-1-carboxamide (840.0 mg, 1.69 mmol) in CH2Cl2 (20.0 mL) was added tert-butyl piperazine-1-carboxylate (631.3 mg, 3.39 mmol) and NaBH3CN (213.0 mg, 3.39 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was quenched with CH3OH. The resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (20/80, v/v) to afford tert-butyl 4-((cis-2-((3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)carbamoyl)cyclopropyl)methyl)piperazine-1-carboxylate (660.0 mg, 58%) as a white solid. LCMS (ESI, m/z): [M+H]+=666.4.
To a solution of tert-butyl 4-((cis-2-((3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)carbamoyl)cyclopropyl)methyl)piperazine-1-carboxylate (75.0 mg, 0.16 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in ACN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was separated by Prep-Chiral-HPLC with the following conditions (Column: Lux 5 um Cellulose-2, 2.12×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 30% B to 30% B in 24 min; 254 nm) to afford N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(piperazin-1-ylmethyl)cyclopropane-1-carboxamide Enantiomer 1 (5.1 mg, 2%) as a white solid and N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(piperazin-1-ylmethyl)cyclopropane-1-carboxamide Enantiomer 2 (3.2 mg, 1%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 171 and 172 in Table 1.
N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(piperazin-1-ylmethyl)cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1: 15.594 min; LCMS (ESI, m/z): [M+H]+=436.3. 1H NMR (400 MHz, DMSO-d6): δ 11.39 (s, 1H), 10.46 (s, 1H), 7.79 (d, J=8.4 Hz, 1H), 7.52 (d, J=8.8 Hz, 1H), 7.29-7.25 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 3.69 (s, 6H), 2.67-2.62 (m, 3H), 2.47-2.42 (m, 1H), 2.38-2.21 (m, 4H), 2.10-2.06 (m, 1H), 1.36-1.30 (m, 1H), 1.04-0.91 (m, 1H), 0.89-0.82 (m, 1H).
N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(piperazin-1-ylmethyl)cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2: 21.022 min; LCMS (ESI, m/z): [M+H]+=436.3. 1H NMR (400 MHz, DMSO-d6): δ 11.39 (s, 1H), 10.46 (s, 1H), 7.79 (d, J=8.4 Hz, 1H), 7.52 (d, J=8.8 Hz, 1H), 7.27-7.25 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 3.69 (s, 6H), 2.67-2.61 (m, 3H), 2.47-2.30 (m, 4H), 2.10-2.06 (m, 1H), 1.35-1.32 (m, 1H), 0.98-0.91 (m, 1H), 0.89-0.82 (m, 1H)
To a solution of 4-cyclopropoxy-3-iodo-2-methoxypyridine (500.0 mg, 1.72 mmol) in 1, 4-dioxane (20.0 mL)/H2O (5.0 mL) was added K3PO4 (1093.8 mg, 5.15 mmol), 6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-ylboronic acid (561.1 mg, 1.72 mmol) and (AMPhosPdCl2)2 (243.3 mg, 0.34 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 3-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-4-cyclopropoxy-2-methoxypyridine (650.0 mg, 84%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=446.2.
To a solution of 3-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-4-cyclopropoxy-2-methoxypyridine (524.0 mg, 1.2 mmol) in THF (20.0 mL) was added Pd2(dba)3 (107.6 mg, 0.12 mmol), LiHMDS (3.5 mL, 1.0 mol/L) and XPhos (112.0 mg, 0.24 mmol) at room temperature under N2. The resulting mixture was stirred at 60° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (440.0 mg, 87%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=427.2.
To a solution of 3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (200.0 mg, 0.47 mmol) in CH2Cl2 (15.0 mL) was added Pyridine (148.3 mg, 1.88 mmol) and phenyl chloroformate (146.8 mg, 0.94 mmol) at 0° C. The resulting mixture was stirred at 0° C. for 1 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added N1, N1-dimethylpropane-1,3-diamine (335.3 mg, 3.28 mmol) and Pyridine (10.0 mL) at room temperature. The resulting mixture was stirred at 60 C for 3 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (20/1, v/v) to afford 1-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)propyl]urea (140.0 mg, 53%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=555.3.
To a solution of 1-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)propyl]urea (120.0 mg, 0.22 mmol) in CH2Cl2 (8.0 mL) was added TFA (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added NH3·H2O (8.0 mL) and CH3CN (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD Cis Column, 30×150 mm, 5 m; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60.0 mL/min; Gradient: 18% B to 35% B in 8 min; Wave Length: 254 nm) to afford 1-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)propyl]urea (Compound 173) (37.4 mg, 39%) as a white solid. LCMS (ESI, m/z): [M+H]+=425.4. 1H NMR (400 MHz, DMSO-d6): δ 11.43 (s, 1H), 9.11 (s, 1H), 8.53 (s, 1H); 8.06 (d, J=6.0 Hz, 1H), 7.49 (d, J=8.4 Hz, 1H), 7.23 (d, J=2.4 Hz, 1H), 7.13 (d, J=5.6 Hz, 1H), 6.95 (d, J=8.4 Hz, 1H), 3.94-3.90 (m, 1H), 3.81 (s, 3H), 3.27-3.22 (m, 2H), 2.29-2.26 (m, 2H), 2.15 (s, 6H), 1.69-1.62 (m, 2H), 0.81-0.78 (m, 2H), 0.66-0.61 (m, 2H).
To a solution of 3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (600.0 mg, 1.40 mmol) in CH2Cl2 (6.0 mL) was added phenyl chloroformate (263.7 mg, 1.68 mmol) and pyridine (444.0 mg, 5.61 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure. To the residue in pyridine (6.0 mL) was added N1,N1-dimethylpropane-1,3-diamine (430.2 mg, 4.21 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography with CH2Cl2/CH3OH (90/10, v/v) to afford 1-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)propyl]urea (390.0 mg, 50%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=556.3.
To a solution of 1-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)propyl]urea (200.0 mg, 0.36 mmol) in CH2Cl2 (4.0 mL) was added TFA (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (4.0 mL) was added NH3·H2O (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 m; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 18% B to 38% B in 8 min, 38% B; Wave Length: 254 nm) to afford 1-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)propyl]urea (Compound 174) (56.7 mg, 36%) as a white solid. LCMS (ESI, m/z): [M+H]+=426.2. 1H NMR (400 MHz, DMSO-d6): δ 11.56 (s, 1H), 9.15 (s, 1H), 8.48 (s, 2H), 7.58 (d, J=8.4 Hz, 1H), 7.33 (s, 1H), 6.98 (d, J=8.8 Hz, 1H), 4.38-4.33 (m, 1H), 3.91 (s, 3H), 3.26-3.21 (m, 2H), 2.29-2.25 (m, 2H), 2.14 (s, 6H), 1.69-1.62 (m, 2H), 0.77-0.74 (m, 2H), 0.69-0.59 (m, 2H).
To a solution of 4,5-dibromo-2H-pyridazin-3-one (5.0 g, 19.69 mmol) in CH3OH (80.0 mL) was added NaOMe (3.2 g, 59.08 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction solution was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with ACN/H2O (1/10, v/v) to afford 4-bromo-5-methoxy-2H-pyridazin-3-one (150.0 mg, 4%) as a white solid. LCMS (ESI, m/z): [M+H]+=205.0.
To a solution of 4-bromo-5-methoxy-2H-pyridazin-3-one (2.0 g, 9.76 mmol) in CHCl3 (30.0 mL) was added Ag2CO3 (10.8 g, 39.02 mmol) and CH3I (11.1 g, 78.04 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with ACN/H2O (1/2, v/v) to afford 4-bromo-3,5-dimethoxypyridazine (100.0 mg, 4%) as a white solid. LCMS (ESI, m/z): [M+H]+=219.0.
To a solution of 4-bromo-3,5-dimethoxypyridazine (130.0 mg, 0.59 mmol) in dioxane/H2O (10.0 mL/2.0 mL) was added 6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-ylboronic acid (193.9 mg, 0.59 mmol), K2CO3 (246.1 mg, 1.78 mmol) and Pd(dppf)Cl2 (43.4 mg, 0.06 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 4-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-3,5-dimethoxypyridazine (70.0 mg, 28%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=421.1.
To a solution of 4-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-3,5-dimethoxypyridazine (80.0 mg, 0.19 mmol) in t-BuOH (6.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (97.9 mg, 0.95 mmol), X-Phos (18.1 mg, 0.04 mmol), K2CO3 (78.8 mg, 0.57 mmol) and Pd(OAc)2 (4.3 mg, 0.02 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h under N2. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with ACN/H2O (1/2, v/v) to afford (1S,2S)—N-[3-(3,5-dimethoxypyridazin-4-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (30.0 mg, 32%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=488.2.
To a solution of (1S,2S)—N-[3-(3,5-dimethoxypyridazin-4-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (50.0 mg, 0.12 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column (YMC-Actus Triart C18 ExRS, 30×150 mm, 5 m; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 15% B to 40% B in 8 min; 254 nm) to afford (1S,2S)—N-[3-(3,5-dimethoxypyridazin-4-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (Compound 175) (6.6 mg, 17%) as a white solid. LCMS (ESI, m/z): [M+H]+=358.2. 1H NMR (400 MHz, DMSO-d6): δ 11.84 (s, 1H), 10.64 (s, 1H), 9.01 (s, 1H), 7.90 (d, J=8.8 Hz, 1H), 7.77 (d, J=8.8 Hz, 1H), 7.64 (s, 1H), 5.01-4.84 (m, 1H), 4.04 (s, 3H), 3.99 (s, 3H), 2.26-2.22 (m, 1H), 1.70-1.63 (m, 1H), 1.19-1.13 (m, 1H).
To a solution of 3,5-dimethoxyaniline (10.0 g, 6.52 mmol) in ACN (10.0 mL) was added Boc2O (21.3 g, 97.93 mmol) and TEA (19.8 g, 195.84 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford tert-butyl N-(3,5-dimethoxyphenyl)carbamate (5.0 g, 30%) as a white solid. LCMS (ESI, m/z): [M+H]+=254.1.
To a solution of Selectfluor (5.0 g, 15.72 mmol) in ACN (50.0 mL) was added tert-butyl N-(3,5-dimethoxyphenyl)carbamate (4.0 g, 15.72 mmol) at 0° C. The resulting mixture was stirred at 0° C. for 2 h. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford tert-butyl N-(2-fluoro-3,5-dimethoxyphenyl)carbamate (1.2 g, 28%) as a white solid. LCMS (ESI, m/z): [M+H]+=272.1.
To a solution of tert-butyl N-(2-fluoro-3,5-dimethoxyphenyl)carbamate (1.0 g, 3.66 mmol) in DCM (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O. The pH value of the mixture was adjusted to 7 with aq. NaHCO3. The mixture was extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 2-fluoro-3,5-dimethoxyaniline (500.0 mg, 79%) as a red solid. LCMS (ESI, m/z): [M+H]+=172.1.
To a solution of 2-fluoro-3,5-dimethoxyaniline (500.0 mg, 2.91 mmol) in propan-2-one (5.0 mL) was added benzoyl isothiocyanate (441.6 mg, 2.91 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 12 h. After the reaction was completed, the resulting mixture was filtered. The solid was collected and dried to afford 1-benzoyl-3-(2-fluoro-3,5-dimethoxyphenyl)thiourea (500.0 mg, crude) as a red oil. LCMS (ESI, m/z): [M+H]+=335.1.
To a solution of 1-benzoyl-3-(2-fluoro-3,5-dimethoxyphenyl)thiourea (500.0 mg, crude) in MeOH (5.0 mL) and H2O (5.0 mL) was added NaOH (39.6 mg, 1.65 mmol). The resulting mixture was stirred at 80° C. for 12 h. After the reaction was completed, the resulting mixture was filtered. The solid was washed with CH3OH and then dried to afford 2-fluoro-3,5-dimethoxyphenylthiourea (300.0 mg, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=231.1.
To a solution of 2-fluoro-3,5-dimethoxyphenylthiourea (300.0 mg, 1.34 mmol) in CHCl3 (5.0 mL) was added Br2 (417.4 mg, 1.5 mmol). The resulting mixture was stirred at 60° C. for 2 h. After the reaction was completed, the resulting mixture was filtered. The solid was collected and dried to afford 4-fluoro-5,7-dimethoxy-1,3-benzothiazol-2-amine (250.0 mg, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=229.0.
To a solution of 4-fluoro-5,7-dimethoxy-1,3-benzothiazol-2-amine (250.0 mg, 1.10 mmol) in THE (5.0 mL) was added NBS (300.0 mg, 1.64 mmol). The resulting mixture was stirred at 60° C. for 2 h. After the reaction was completed, the resulting mixture was filtered. The solid was collected and dried to afford 6-bromo-4-fluoro-5,7-dimethoxy-1,3-benzothiazol-2-amine (250.0 mg, crude) as an off-white solid. LCMS (ESI, m/z): [M+H]+=306.9.
To a solution of 6-bromo-4-fluoro-5,7-dimethoxy-1,3-benzothiazol-2-amine (250.0 mg, crude) in THF (5.0 mL) was added tert-butyl nitrite (125.9 mg, 1.36 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with dichloromethane. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 6-bromo-4-fluoro-5,7-dimethoxy-1,3-benzothiazole (200.0 mg, 84%) as a white solid. LCMS (ESI, m/z): [M+H]+=291.9.
To a solution of 6-bromo-4-fluoro-5,7-dimethoxy-1,3-benzothiazole (200.0 mg, 0.69 mmol) in dioxane (5.0 mL) and H2O (0.5 mL) was added 6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-ylboronic acid (268.4 mg, 0.82 mmol), Pd(dppf)Cl2 (50.2 mg, 0.07 mmol) and K2CO3 (283.9 mg, 2.07 mmol) under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 6-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-4-fluoro-5,7-dimethoxy-1,3-benzothiazole (200.0 mg, 59%) as a green oil. LCMS (ESI, m/z): [M+H]+=494.1.
To a solution of 6-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-4-fluoro-5,7-dimethoxy-1,3-benzothiazole (200.0 mg, 0.45 mmol) in dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (125.2 mg, 1.25 mmol), K2CO3 (167.8 mg, 1.25 mmol), BrettPhos (43.4 mg, 0.08 mmol) and BrettPhos Pd G3 (36.7 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(4-fluoro-5,7-dimethoxy-1,3-benzothiazol-6-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (100.0 mg, 44%) as a green oil. LCMS (ESI, m/z): [M+H]+=561.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(4-fluoro-5,7-dimethoxy-1,3-benzothiazol-6-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (100.0 mg, 0.18 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 49% B to 63% B in 10 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-(4-fluoro-5,7-dimethoxybenzo[d]thiazol-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (Compound 176) (2.3 mg, 2%) as a white solid. LCMS (ESI, m/z): [M+H]+=431.1. 1H NMR (300 MHz, DMSO-d6): δ 11.79 (s, 1H), 10.67 (s, 1H), 9.42 (s, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.76 (d, J=8.7 Hz, 1H), 7.55 (d, J=2.4 Hz, 1H), 5.03-4.79 (m, 1H), 3.61 (s, 3H), 3.47 (s, 3H), 2.26-2.20 (m, 1H), 1.68-1.59 (m, 1H), 1.17-1.08 (m, 1H).
To a solution of ethyl (1R,5S,6R)-3-azabicyclo[3.1.0]hexane-6-carboxylate hydrochloride (200.0 mg, 1.04 mmol) in THE (5.0 mL) was added formaldehyde (522.2 mg, 5.22 mmol) and NaBH(OAc)3 (442.3 mg, 2.09 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was quenched with MeOH. The resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (90/10, v/v) to afford ethyl (1R,5S,6R)-3-methyl-3-azabicyclo[3.1.0]hexane-6-carboxylate (140.0 mg, 79%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=170.1.
To a mixture of ethyl (1R,5S,6R)-3-methyl-3-azabicyclo[3.1.0]hexane-6-carboxylate (120.0 mg, 0.71 mmol) and 3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (141.7 mg, 0.35 mmol) in THE (3.0 mL) was added dropwise AlMe3 (1.8 mL, 2 mol/L) at 0° C. The mixture was stirred at 80° C. for 2 h. After the reaction was completed, the reaction mixture was quenched with water. The resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (90/10, v/v) to afford (1R,5S,6R)—N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-3-azabicyclo[3.1.0]hexane-6-carboxamide (80.0 mg, 21%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=523.3.
To a solution of (1R,5S,6R)—N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-3-azabicyclo[3.1.0]hexane-6-carboxamide (60.0 mg, 0.12 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL). The mixture was stirred at room temperature for 1 h. The mixture was concentrated under vacuum. To the above residue were added ACN (2.0 mL) and NH3·H2O (2.0 mL). The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18 ExRS, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 34% B to 48% B in 8 min; 254 nm) to afford (1R,5S,6R)—N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-methyl-3-azabicyclo[3.1.0]hexane-6-carboxamide (Compound 177) (15.5 mg, 34%) as a white solid. LCMS (ESI, m/z): [M+H]+=393.2. H NMR (300 MHz, DMSO-d6): δ 11.42 (s, 1H), 10.35 (s, 1H), 7.77 (d, J=8.4 Hz, 1H), 7.52-7.50 (m, 1H), 7.29-7.25 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 3.69 (s, 6H), 3.00-2.98 (m, 2H), 2.36-2.31 (m, 3H), 2.25 (s, 3H), 1.85 (s, 2H).
To a solution of 4-cyclopropoxy-3-iodo-2-methoxypyridine (268.0 mg, 0.92 mmol) in 1,4-dioxane/H2O (8.0 mL/1.6 mL) was added 6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-ylboronic acid (300.8 mg, 0.92 mmol), K3PO4 (390.9 mg, 1.84 mmol) and PdAMPhos (130.4 mg, 0.18 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 3-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-4-cyclopropoxy-2-methoxypyridine (253.0 mg, 61%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=446.2.
To a solution of 3-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-4-cyclopropoxy-2-methoxypyridine (170.0 mg, 0.38 mmol) in dioxane (6.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (117.9 mg, 1.14 mmol), K2CO3 (158.0 mg, 1.14 mmol), Brettphos (40.9 mg, 0.08 mmol) and Brettphos Pd G3 (34.6 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)—N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (80.0 mg, 41%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=513.2.
To a solution of (1S,2S)—N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (80.0 mg, 0.16 mmol) in DCM (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added NH3·H2O (3.0 mL) and ACN (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for additional 4 h. After the reaction was completed, the mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (4/1, v/v) and then purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 10% B to 40% B in 8 min, 254 nm) to afford (1S,2S)—N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (Compound 178) (10.3 mg, 16%) as a white solid. LCMS (ESI, m/z): [M+H]+=383.1. 1H NMR (400 MHz, DMSO-d6): δ 11.55 (s, 1H), 10.61 (s, 1H), 8.08 (d, J=6.0 Hz, 1H), 7.84 (d, J=8.4 Hz, 1H), 7.58 (d, J=8.4 Hz, 1H), 7.36 (d, J=2.4 Hz, 1H), 7.14 (d, J=5.6 Hz, 1H), 5.02-4.83 (m, 1H), 3.95-3.92 (m, 1H), 3.82 (s, 3H), 2.28-2.19 (m, 1H), 1.72-1.55 (m, 1H), 1.16-1.13 (m, 1H), 0.80-0.76 (m, 2H), 0.66-0.61 (m, 2H).
To a solution of 3-{5-methoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (170.0 mg, 0.40 mmol) in CH2Cl2 (5.0 mL) was added pyridine (125.8 mg, 1.59 mmol) and phenyl chloroformate (74.7 mg, 0.48 mmol) at 0° C. The mixture was stirred at room temperature for 1 h. The mixture was concentrated under vacuum. To the above residue was added pyridine (3.0 mL) and N1,N1-dimethylpropane-1,3-diamine (121.9 mg, 1.19 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 3 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (85/15, v/v) to afford 3-[3-(dimethylamino)propyl]-1-(3-{5-methoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl)urea (100.0 mg, 45%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=556.2.
To a solution of 3-[3-(dimethylamino)propyl]-1-(3-{5-methoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl)urea (80.0 mg, 0.14 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. To the above residue were added ACN (2.0 mL) and NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18 ExRS, 30×250 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 20% B to 30% B in 11 min; 254 nm) to afford 3-[3-(dimethylamino)propyl]-1-(3-{5-methoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1H-pyrrolo[2,3-b]pyridin-6-yl)urea (Compound 179) (19.8 mg, 32%) as a white solid. LCMS (ESI, m/z): [M+H]+=426.3. 1H NMR (400 MHz, DMSO-d6): δ 11.71 (s, 1H), 9.51 (s, 1H), 9.19 (s, 1H), 8.78 (s, 1H), 8.45 (s, 1H), 8.16 (d, J=8.4 Hz, 1H), 7.73 (d, J=2.4 Hz, 1H), 7.12 (d, J=8.8 Hz, 1H), 4.06 (s, 3H), 3.27-3.22 (m, 2H), 2.34-2.27 (m, 2H), 2.14 (s, 6H), 1.70-1.64 (m, 2H).
The cis-N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluoro-2-(hydroxymethyl)cyclopropanecarboxamide (94.0 mg, 0.24 mmol) was separated by CHIRAL-HPLC with the following conditions (Column: CHIRAL ART Amylose-SA, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 50% B to 50% B in 17 min; Wave Length: 220/254 nm) to afford N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluoro-2-(hydroxymethyl)cyclopropanecarboxamide Enantiomer 1 (18.2 mg, 19%) as a white solid and N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluoro-2-(hydroxymethyl)cyclopropanecarboxamide Enantiomer 2 (22.6 mg, 24%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 180 and 181 in Table 1.
N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluoro-2-(hydroxymethyl)cyclopropanecarboxamide Enantiomer 1: Retention Time 1=7.02 min; LCMS (ESI, m/z): [M+H]+=386.1. 1H NMR (400 MHz, DMSO-d6): δ 11.43 (d, J=1.6 Hz, 1H), 10.43 (s, 1H), 7.82 (d, J=8.4 Hz, 1H), 7.55 (d, J=8.8 Hz, 1H), 7.30-7.26 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 5.15-5.12 (m, 1H), 3.72-3.66 (m, 8H), 2.33-2.29 (m, 1H), 1.76-1.68 (m, 1H), 1.25-1.22 (m, 1H).
N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluoro-2-(hydroxymethyl)cyclopropanecarboxamide Enantiomer 2: Retention Time 2=14.64 min; LCMS (ESI, m/z): [M+H]+=386.3. H NMR (400 MHz, DMSO-d6): δ 11.41 (s, 1H), 10.39 (s, 1H), 7.81 (d, J=8.0 Hz, 1H), 7.55 (d, J=8.4 Hz, 1H), 7.30-7.26 (m, 2H), 6.75 (d, J=8.0 Hz, 2H), 5.13-5.10 (m, 1H), 3.73-3.66 (m, 8H), 2.33-2.29 (m, 1H), 1.77-1.69 (m, 1H), 1.29-1.18 (m, 1H).
To a solution of 6-chloro-3-iodo-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridine (400.0 mg, 0.98 mmol) in dioxane/H2O (10.0 mL/2.0 mL) was added 2-methoxy-6-methylphenylboronic acid (162.0 mg, 0.97 mmol), K2CO3 (404.8 mg, 2.93 mmol) and Pd(dppf)Cl2 (71.4 mg, 0.10 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 6-chloro-3-(2-methoxy-6-methylphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridine (120.0 mg, 30%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=404.1.
To a solution of 6-chloro-3-(2-methoxy-6-methylphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridine (210.0 mg, 0.52 mmol) in 1,4-dioxane (16.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (267.9 mg, 2.60 mmol), BrettPhos (55.8 mg, 0.10 mmol), Cs2CO3 (508.1 mg, 1.56 mmol) and BrettPhos Pd G3 (47.1 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)-2-fluoro-N-[3-(2-methoxy-6-methylphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (120.0 mg, 49%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=471.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(2-methoxy-6-methylphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (120.0 mg 0.12 mmol) in CH2Cl2 (10.0 mL) was added TFA (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (8.0 mL) was added NH3·H2O (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 33% B to 43% B in 8 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(2-methoxy-6-methylphenyl)-1H-pyrazolo[3,4-b]pyridin-6-yl]cyclopropane-1-carboxamide (Compound 182) (16.9 mg, 14%) as a white solid. LCMS (ESI, m/z): [M+H]+=341.2. 1H NMR (400 MHz, DMSO-d6): δ 13.39 (s, 1H), 10.96 (s, 1H), 7.93 (d, J=8.8 Hz, 1H), 7.78 (d, J=8.8 Hz, 1H), 7.36-7.32 (m, 1H), 7.00-6.94 (m, 2H), 5.05-4.85 (m, 1H), 3.66 (s, 3H), 2.28-2.25 (m, 1H), 2.12 (s, 3H), 1.70-1.62 (m, 1H), 1.23-1.16 (m, 1H).
To a solution of 3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (300.0 mg, 0.75 mmol) in THE (10.0 mL) was added trans-ethyl 2-((tert-butyldiphenylsilyloxy)methyl)-2-fluorocyclopropanecarboxylate (902 mg, 2.25 mmol) and AlMe3 (1.9 mL, 2 mol/L) at 0° C. under N2. The resulting mixture was stirred at 80° C. for 5 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (30/70, v/v) to afford trans-2-((tert-butyldiphenylsilyloxy)methyl)-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropanecarboxamide (200.0 mg, 35%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=754.3.
The solution of trans-2-((tert-butyldiphenylsilyloxy)methyl)-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropanecarboxamide (150.0 mg, 0.20 mmol) in HCl/dioxane (5.0 mL, 4 mol/L) was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. To the residue in ACN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse flash column chromatography with petroleum MeOH/H2O (70/30, v/v) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluoro-2-(hydroxymethyl)cyclopropanecarboxamide (Compound 183) (1.2 mg, 2%) as a white solid. LCMS (ESI, m/z): [M+H]+=386.3. 1H NMR (400 MHz, DMSO-d6): δ 11.47 (s, 1H), 10.64 (s, 1H), 7.80 (d, J=8.4 Hz, 1H), 7.55 (d, J=8.8 Hz, 1H), 7.30-7.26 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 5.06-5.03 (m, 1H), 3.98-3.91 (m, 1H), 3.86-3.74 (m, 1H) 3.69 (s, 6H) 2.69-2.62 (m, 1H), 1.53-1.47 (m, 1H), 1.35-1.30 (m, 1H).
To a mixture of piperidine-4-carbonitrile (1.0 g, 9.07 mmol) and K2CO3 (3.7 g, 27.20 mmol) in MeCN (10.0 mL) was added bromoethane (1.0 g, 9.08 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (5/1, v/v) to afford 1-ethylpiperidine-4-carbonitrile (750.0 mg, 59%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=139.1
To a solution of 1-ethylpiperidine-4-carbonitrile (2.2 g, 15.91 mmol) in THE (20.0 mL) was added LiHMDS (31.0 mL, 1 mol/L) at −10° C. under N2. The resulting mixture was stirred at −10° C. for 1 h under N2. Then 2-bromoethoxy(tert-butyl)dimethylsilane (7.6 g, 31.85 mmol) was added dropwise to the mixture at −10° C. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 4-[2-[(tert-butyldimethylsilyl)oxy]ethyl]-1-ethylpiperidine-4-carbonitrile (3.0 g, 63%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=297.2
The solution of 4-[2-[(tert-butyldimethylsilyl)oxy]ethyl]-1-ethylpiperidine-4-carbonitrile (3.0 g, 10.11 mmol) in HCl/1,4-dioxane (20.0 mL, 4 mol/L) was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was evaporated in vacuo. The PH of the residue was adjusted to 8 with NaHCO3 (aq) and then extracted with ethyl acetate. The aqueous layer was concentrated under reduced pressure to afford 1-ethyl-4-(2-hydroxyethyl)piperidine-4-carbonitrile (2.3 g, crude) as yellow solid. LCMS (ESI, m/z): [M+H]+=183.1
To a mixture of 1-ethyl-4-(2-hydroxyethyl)piperidine-4-carbonitrile (300.0 mg, 1.64 mmol) and PPh3 (863.4 mg, 3.29 mmol) in THE (5.0 mL) was added DIAD (665.6 mg, 3.29 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 15 min under N2. Then DPPA (905.9 mg, 3.29 mmol) was added to the mixture at 0° C. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) to afford 4-(2-azidoethyl)-1-ethylpiperidine-4-carbonitrile (300.0 mg, 87%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=208.1
To a solution of 4-(2-azidoethyl)-1-ethylpiperidine-4-carbonitrile (1.4 g, 6.75 mmol) in THE (20.0 mL) was added triphenylphosphine (2.6 g, 10.13 mmol) at room temperature. The resulting mixture was stirred at 70° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The aqueous layer was concentrated under reduced pressure to afford 4-(2-aminoethyl)-1-ethylpiperidine-4-carbonitrile (1.1 g, crude) as a yellow oil. LCMS (ESI, m/z): [M+H]+=182.2
To a mixture of 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (1.0 g, 2.75 mmol) and pyridine (872.6 mg, 11.03 mmol) in DCM (10.0 mL) was added phenyl chloroformate (518.2 mg, 3.31 mmol) at 0° C. The resulting mixture was stirred for at room temperature 2 h. The resulting mixture was concentrated under reduced pressure. To the above mixture was added pyridine (10.0 mL) and 4-(2-aminoethyl)-1-ethylpiperidine-4-carbonitrile (500.0 mg, 2.75 mmol) at room temperature. The resulting mixture was stirred at 70° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (8/1, v/v) to afford 1-[2-(4-cyano-1-ethylpiperidin-4-yl)ethyl]-3-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (450.0 mg, 28%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=577.3
To a solution of 1-[2-(4-cyano-1-methylpiperidin-4-yl)ethyl]-3-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (450.0 mg, 0.80 mmol) in DCM (5.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was concentrated under reduced pressure. To the above mixture was added NH3·H2O (2.0 mL) and ACN (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 11% B to 20% B in 8 min, 254/220 nm) to afford 1-[2-(4-cyano-1-ethylpiperidin-4-yl)ethyl]-3-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]urea (Compound 184) (77.3 mg, 22%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=447.2. 1H NMR (300 MHz, DMSO-d6): δ 11.41 (s, 1H), 9.42 (s, 1H), 7.99-7.89 (m, 2H), 7.56-7.52 (m, 2H), 7.30-7.24 (m, 1H), 7.11 (d, J=7.5 Hz, 1H), 7.05-7.00 (m, 1H), 3.82 (s, 3H), 3.62-3.48 (m, 2H), 3.34-2.73 (m, 3H), 2.08-1.76 (m, 6H), 1.29-1.19 (m, 3H).
The cis-N-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (Compound 41) (33.0 mg, 0.08 mmol) was separated by CHIRAL-HPLC with the following conditions (Column: CHIRALPAK IH, 2×25 cm, 5 um; Mobile Phase A: MTBE (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 10% B to 10% B in 22 min; Wave Length: 254/220 nm) to afford N-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 1 (10.4 mg, 31%) as a white solid and N-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 2 (10.0 mg, 30%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 185 and 186 in Table 1.
N-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1: 7.772 min; LCMS (ESI, m/z): [M+H]+=420.3. 1H NMR (300 MHz, DMSO-d6): δ 11.53 (d, J=2.1 Hz, 1H), 10.51 (s, 1H), 7.97 (d, J=8.7 Hz, 1H), 7.88 (d, J=8.7 Hz, 1H), 7.56-7.52 (m, 2H), 7.30-7.25 (m, 1H), 7.11 (d, J=7.5 Hz, 1H), 7.05-7.00 (m, 1H), 3.82 (s, 3H), 2.60-2.52 (m, 2H), 2.51-2.46 (m, 1H), 2.44-2.32 (m, 3H), 2.31-2.17 (m, 4H), 2.16-2.10 (m, 1H), 2.08 (s, 3H), 1.35-1.30 (m, 1H), 1.03-0.99 (m, 1H), 0.91-0.82 (m, 1H).
N-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2:17.229 min; LCMS (ESI, m/z): [M+H]+=420.3. 1H NMR (300 MHz, DMSO-d6): δ 11.53 (d, J=1.8 Hz, 1H), 10.51 (s, 1H), 7.97 (d, J=8.7 Hz, 1H), 7.88 (d, J=8.7 Hz, 1H), 7.56-7.52 (m, 2H), 7.30-7.25 (m, 1H), 7.11 (d, J=7.5 Hz, 1H), 7.05-7.00 (m, 1H), 3.82 (s, 3H), 2.60-2.52 (m, 2H), 2.51-2.46 (m, 1H), 2.44-2.32 (m, 3H), 2.31-2.17 (m, 4H), 2.16-2.10 (m, 1H), 2.08 (s, 3H), 1.38-1.31 (m, 1H), 1.03-0.99 (m, 1H), 0.91-0.82 (m, 1H).
To a solution of 3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (450.0 mg, 1.13 mmol) in DCM (10.0 mL) was added pyridine (356.3 mg, 4.50 mmol) and phenyl chloroformate (211.6 mg, 1.35 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. To the residue in pyridine (10 mL) was added tert-butyl 3-(aminomethyl)-3-fluoroazetidine-1-carboxylate (690.1 mg, 3.38 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 4 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) to afford tert-butyl 3-[({[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}amino)methyl]-3-fluoroazetidine-1-carboxylate (600.0 mg, 85%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=630.3.
To a solution of tert-butyl 3-[({[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}amino)methyl]-3-fluoroazetidine-1-carboxylate (580.0 mL, 0.92 mmol) in DCM (5.0 mL) was added HCOOH (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the pH value of the mixture was adjusted to 8 with NaHCO3 (aq). The mixture was extracted with EtOAc. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 1-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[(3-fluoroazetidin-3-yl)methyl]urea (420.0 mg, crude) as a yellow oil. LCMS (ESI, m/z): [M+H]+=530.3.
To a solution of 1-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[(3-fluoroazetidin-3-yl)methyl]urea (420.0 mg, crude) in THF/CH3OH (20.0 mL/4.0 mL) was added NaBH3CN (149.5 mg, 2.38 mmol) and HCHO (238.1 mg, 7.93 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 1-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[(3-fluoro-1-methylazetidin-3-yl)methyl]urea (400.0 mg, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=544.3.
To a solution of 1-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[(3-fluoro-1-methylazetidin-3-yl)methyl]urea (400.0 mg, crude) in CH2Cl2 (20.0 mL) was added TFA (20.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (20.0 mL) was added NH3·H2O (20.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 23% B to 53% B in 9 min; 254 nm) to afford 1-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[(3-fluoro-1-methylazetidin-3-yl)methyl]urea (Compound 187) (12.9 mg, 4%) as a white solid. LCMS (ESI, m/z): [M+H]+=414.2. 1H NMR (400 MHz, DMSO-d6): δ 11.40 (s, 1H), 9.25 (s, 1H), 8.76 (s, 1H), 7.49 (d, J=8.4 Hz, 1H), 7.28-7.24 (m, 1H), 7.17 (d, J=2.4 Hz, 1H), 6.97 (d, J=8.4 Hz, 1H), 6.73 (d, J=8.4 Hz, 2H), 3.75-3.67 (m, 10H), 3.46-3.31 (m, 2H), 2.45 (s, 3H).
To a solution of LiAlH4 (5.5 g, 114.12 mmol) in Et2O (100.0 mL) was added AlCl3 (6.4 g, 48.04 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 30 min. Then methyl 2-fluoroacrylate (5.0 g, 48.04 mmol) was added dropwise to the mixture at 0° C. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the resulting mixture was filtered. To the above filtrate was added TBDPSCl (19.8 g, 72.12 mmol) and 1H-imidazole (11.3 g, 96.15 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (95/5, v/v) to afford tert-butyl(2-fluoroallyloxy)diphenylsilane (4.0 g, 19%) as a yellow oil.
To a solution of tert-butyl(2-fluoroallyloxy)diphenylsilane (1.0 g, 3.18 mmol) in DCM (5.0 mL) was added dropwise ethyl 2-diazoacetate (1.1 g, 9.54 mmol) at 0° C. for 8 h. The resulting mixture was stirred at room temperature for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to afford cis-ethyl 2-((tert-butyldiphenylsilyloxy)methyl)-2-fluorocyclopropanecarboxylate (300.0 mg, 24%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=401.2. and trans-ethyl 2-((tert-butyldiphenylsilyloxy)methyl)-2-fluorocyclopropanecarboxylate. (400.0 mg, 31%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=401.2.
To a solution of 3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (400.0 mg, 1.00 mmol) in THE (10.0 mL) was added cis-ethyl 2-((tert-butyldiphenylsilyloxy)methyl)-2-fluorocyclopropanecarboxylate (1.2 g, 3.00 mmol) and AlMe3 (2.5 mL, 2 mol/L) at 0° C. under N2. The resulting mixture was stirred at 80° C. for 5 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (30/70, v/v) to afford cis-2-((tert-butyldiphenylsilyloxy)methyl)-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropanecarboxamide (150.0 mg, 20%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=754.3.
The solution of cis-2-((tert-butyldiphenylsilyloxy)methyl)-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluorocyclopropanecarboxamide (90.0 mg, 0.07 mmol) in HCl/dioxane (5.0 mL, 4 mol/L) was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. To the residue in ACN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse flash column chromatography with petroleum MeOH/H2O (70/30, v/v) to afford cis-N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-fluoro-2-(hydroxymethyl)cyclopropanecarboxamide (Compound 188) (4.2 mg, 9%) as a white solid. LCMS (ESI, m/z): [M+H]+=386.2. 1H NMR (400 MHz, DMSO-d6): δ 11.41 (d, J=2.0 Hz, 1H), 10.40 (s, 1H), 7.81 ((d, J=8.4 Hz, 1H), 7.55 ((d, J=8.8 Hz, 1H), 7.30-7.26 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 5.13-5.10 (m, 1H), 3.73-3.66 (m, 8H), 2.34-2.29 (m, 1H), 1.77-1.70 (m, 1H), 1.29-1.08 (m, 1H).
To a solution of 5-bromo-4-cyclopropoxy-6-methoxypyrimidine (500.0 mg, 2.04 mmol) in 1,4-dioxane (10.0 mL) was added 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (1.6 g, 6.12 mmol), KOAc (400.5 mg, 4.08 mmol) and Pd(dppf)Cl2 (149.3 mg, 0.20 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 4-cyclopropoxy-6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine (240.0 mg, 40%) as a light brown oil. LCMS (ESI, m/z): [M+H]+=293.2.
To a solution of 4-cyclopropoxy-6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine (320.0 mg, 1.10 mmol) in 1,4-dioxane/H2O (30.0 mL/6.0 mL) was add 6-chloro-3-iodo-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridine (372.5 mg, 0.91 mmol), K2CO3 (302.8 mg, 2.19 mmol) and Pd(PPh3)4 (126.6 mg, 0.11 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (7/1, v/v) to afford 5-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridin-3-yl)-4-cyclopropoxy-6-methoxypyrimidine (295.0 mg, 60%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=448.1.
To a solution of 5-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridin-3-yl)-4-cyclopropoxy-6-methoxypyrimidine (255.0 mg, 0.57 mmol) in t-BuOH (25.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (293.4 mg, 2.85 mmol), XPhos (54.3 mg, 0.11 mmol), K2CO3 (236.0 mg, 1.71 mmol) and Pd(OAc)2 (12.8 mg, 0.06 mmol) at room temperature under N2. The final reaction mixture was stirred at 100° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/2, v/v) to afford (1S,2S)—N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (85.0 mg, 29%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=515.2.
To a solution of (1S,2S)—N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (85.0 mg, 0.17 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (3.0 mL) was added NH3·H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 35% B in 9 min; 254/220 nm) to afford (1S,2S)—N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (Compound 189) (13.5 mg, 21%) as a white solid. LCMS (ESI, m/z): [M+H]+=385.2. 1H NMR (400 MHz, DMSO-d6): δ 13.55 (s, 1H), 11.01 (s, 1H), 8.64 (s, 1H), 7.98-7.90 (m, 2H), 5.04-4.87 (m, 1H), 4.37-4.34 (m, 1H), 3.89 (s, 3H), 2.28-2.21 (m, 1H), 1.70-1.63 (m, 1H), 1.22-1.16 (m, 1H), 0.77-0.72 (m, 2H), 0.57-0.52 (m, 2H).
To a solution of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-formylcyclopropane-1-carboxamide (500.0 mg, 1.01 mmol) in CH2Cl2 (20.0 mL) was added dimethylamine hydrochloride (246.8 mg, 3.03 mmol) and NaBH3CN (190.2 mg, 3.03 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was quenched with CH3OH. The resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (80/20, v/v) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (210.0 mg, 39%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=525.3.
To a solution of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (210.0 mg, 0.40 mmol) in DMF (5.0 mL) was added ethylenediamine (120.3 mg, 2.00 mmol) and TBAF (313.9 mg, 1.20 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (75/25, v/v) and then separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK ID, 2×25 cm, 5 um; Mobile Phase A: Hex (0.2% IPA)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 30% to 30% in 23 min; 220/254 nm) to afford N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide Enantiomer 1 (19.8 mg, 12%) as a white solid and N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide Enantiomer 2 (33.3 mg, 21%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 190 and 191 in Table 1.
N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1:14.6 min; LCMS (ESI, m/z): [M+H]+=395.2. H NMR (300 MHz, DMSO-d6): δ 11.42 (s, 1H), 10.52 (s, 1H), 7.81 (d, J=8.7 Hz, 1H), 7.53 (d, J=8.7 Hz, 1H), 7.30-7.25 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 3.69 (s, 6H), 2.32-2.25 (m, 1H), 2.19-2.12 (m, 7H), 1.92-1.86 (m, 1H), 1.36-1.32 (m, 1H), 1.07-1.04 (m, 1H), 0.74-0.65 (m, 1H).
N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2:18.357 min; LCMS (ESI, m/z): [M+H]+=395.2. H NMR (300 MHz, DMSO-d6): δ 11.41 (s, 1H), 10.52 (s, 1H), 7.81 (d, J=8.7 Hz, 1H), 7.53 (d, J=8.4 Hz, 1H), 7.30-7.25 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 3.69 (s, 6H), 2.32-2.25 (m, 1H), 2.19-2.12 (m, 7H), 1.92-1.86 (m, 1H), 1.36-1.31 (m, 1H), 1.07-1.04 (m, 1H), 0.74-0.65 (m, 1H). d
To a solution of (trans-(2-((benzyloxy)methyl)-3-fluorocyclopropyl)methoxy)triisopropylsilane (23.0 g, 62.74 mmol) in THE (200.0 mL) was added TBAF (19.7 g, 75.39 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (40/60, v/v) to afford trans-(2-((benzyloxy)methyl)-3(R)-fluorocyclopropyl)methanol (1.6 g, 12%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=211.1.
To a solution of (CClO)2 (1.9 g, 15.22 mmol) in CH2Cl2 (20.0 mL) was added dropwise a solution of DMSO (4.8 g, 60.88 mmol) in CH2Cl2 (20.0 mL) at −78° C. under N2. The resulting mixture was stirred at −78° C. for 0.5 h under N2. Then a solution of trans-(2-((benzyloxy)methyl)-3(R)-fluorocyclopropyl)methanol (1.6 g, 7.61 mmol) in CH2Cl2 (20.0 mL) was added dropwise to the mixture at −78° C. under N2. The resulting mixture was stirred at −78° C. for 1 h. Then TEA (6.2 g, 60.88 mmol) was added dropwise to the mixture at −78° C. under N2. The resulting mixture was stirred at −78° C. for 0.5 h under N2. After the reaction was completed, the resulting mixture was quenched with aqueous NH4Cl at 0° C. and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to afford trans-(2-((benzyloxy)methyl)-3(S)-fluorocyclopropyl)-1-carbaldehyde (1.1 g, 69%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=209.1.
To a mixture of trans-(2-((benzyloxy)methyl)-3(S)-fluorocyclopropyl)-1-carbaldehyde (1.1 g, 5.28 mmol) and NH2SO3H (5.1 g, 52.83 mmol) in t-BuOH/H2O (2.0/18.0 mL) was added NaClO2 (4.8 g, 52.83 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 1.5 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (50/50, v/v) to afford trans-(2-((benzyloxy)methyl)-3(S)-fluorocyclopropyl)-1-carboxylic acid (800.0 mg, 67%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=225.1.
To a mixture of 3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (1.4 g, 3.57 mmol) and DIEA (1.4 g, 10.70 mmol) in DMF (20.0 mL) was added trans-(2-((benzyloxy)methyl)-3(S)-fluorocyclopropyl)-1-carboxylic acid (800.0 mg, 3.57 mmol) and HATU (1.6 g, 4.28 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (90/10, v/v) to afford trans-2-((benzyloxy)methyl)-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(S)-fluorocyclopropane-1-carboxamide (1.4 g, 65%) as a white solid. LCMS (ESI, m/z): [M+H]+=606.3.
To a solution of trans-2-((benzyloxy)methyl)-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(S)-fluorocyclopropane-1-carboxamide (1.3 g, 2.15 mmol) in CH3OH (15.0 mL) was added Pd/C (228.4 mg, dry) at room temperature under N2. The resulting mixture was stirred at room temperature for 4 h under H2. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with petroleum CH3CN/H2O (28/72, v/v) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(S)-fluoro-2-(hydroxymethyl)cyclopropane-1-carboxamide (570.0 mg, 25%) as a white solid. LCMS (ESI, m/z): [M+H]+=516.2.
To a solution of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(S)-fluoro-2-(hydroxymethyl)cyclopropane-1-carboxamide (570.0 mg, 1.11 mmol) in CH2Cl2 (10.0 mL) was added Dess-Martin (701.1 mg, 1.62 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with aq. NaHCO3, brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(S)-fluoro-2-formylcyclopropane-1-carboxamide (600.0 mg, crude) as a brown solid. LCMS (ESI, m/z): [M+H]+=514.2.
To a mixture of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(S)-fluoro-2-formylcyclopropane-1-carboxamide (600.0 mg, crude) and 1-methylpiperazine (351.0 mg, 3.51 mmol) in CH2C2 (10.0 mL) was added NaBH3CN (330.3 mg, 3.51 mmol) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC with CH2Cl2/CH3OH (20/1, v/v) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(S)-fluoro-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (200.0 mg, 34%) as a brown solid. LCMS (ESI, m/z): [M+H]+=598.3.
To a solution of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(S)-fluoro-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (180.0 mg, 0.30 mmol) in CH2Cl2 (4.0 mL) was added TFA (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (4.0 mL) was added NH3·H2O (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 8 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Shield RP18 OBD Column, 30×150 mm, 5 m; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 50% B in 8 min; Wave Length: 254 nm) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(S)-fluoro-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (Compound 192) (5.2 mg, 4%) as a white solid. LCMS (ESI, m/z): [M+H]+=468.2. 1H NMR (400 MHz, DMSO-d6): δ 11.45 (s, 1H), 10.63 (s, 1H), 7.75 (d, J=8.4 Hz, 1H), 7.54 (d, J=8.8 Hz, 1H), 7.30-7.26 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 4.99-4.82 (m, 1H), 3.69 (s, 6H), 2.73-2.66 (m, 1H), 2.62-2.50 (m, 1H), 2.47-2.26 (m, 8H), 2.09 (s, 3H), 2.06-1.99 (m, 1H).
To a solution of 3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (300.0 mg, 0.75 mmol) in CH2Cl2 (5.0 mL) was added pyridine (224.0 mg, 3.00 mmol) and phenyl chloroformate (140.4 mg, 0.90 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated in vacuo to afford phenyl (3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)carbamate (400.0 mg, crude) as a colorless oil. LCMS (ESI, m/z): [M+H]+=520.2
To a solution of phenyl N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamate (300.0 mg, 0.57 mmol) in pyridine (3.0 mL) was added ethanolamine (42.3 mg, 0.69 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (92/8, v/v) to afford 1-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-(2-hydroxyethyl)urea (250.0 mg, 78%) as a white solid. LCMS (ESI, m/z): [M+H]+=487.2
To a solution of 1-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-(2-hydroxyethyl)urea (100.0 mg, 0.21 mmol) in CH2Cl2 (3.0 mL) was added Dess-Martin (348.6 mg, 0.82 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 1-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-(2-oxoethyl)urea (100.0 mg, crude) as a black solid. LCMS (ESI, m/z): [M+H]+=485.2
To a solution of 1-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-(2-oxoethyl)urea (100.0 mg, crude) in CH2Cl2 (3.0 mL) was added 6-methyl-3,6-diazabicyclo[3.1.1]heptane (69.4 mg, 0.61 mmol) and NaBH3CN (13.0 mg, 0.20 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with petroleum CH3CN/H2O (60/40, v/v) to afford 1-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-(2-{6-methyl-3,6-diazabicyclo[3.1.1]heptan-3-yl}ethyl)urea (30.0 mg, 25%) as a white solid. LCMS (ESI, m/z): [M+H]+=581.3.
To a solution of 1-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-(2-{6-methyl-3,6-diazabicyclo[3.1.1]heptan-3-yl}ethyl)urea (30.0 mg, 0.06 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (3.0 mL) was added NH3·H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 47% B in 9 min; 254 nm) to afford 1-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-(2-{6-methyl-3,6-diazabicyclo[3.1.1]heptan-3-yl}ethyl)urea (Compound 193) (1.4 mg, 6%) as a white solid. LCMS (ESI, m/z): [M+H]+=451.4. H NMR (400 MHz, DMSO-d6): δ 11.27 (s, 1H), 9.17 (s, 1H), 8.49 (s, 1H), 7.46 (d, J=8.4 Hz, 1H), 7.27 (d, J=8.4 Hz, 1H), 7.15 (d, J=2.0 Hz, 1H), 6.97 (d, J=8.4 Hz, 1H), 6.73 (d, J=8.4 Hz, 2H), 3.68 (s, 6H), 2.93-2.83 (m, 4H), 2.76-2.73 (m, 2H), 2.26-2.12 (m, 1H), 2.01 (s, 3H), 1.87-1.82 (m, 1H).
To a solution of 6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-ylboronic acid (1.20 g, 3.67 mmol) in H2O (1.5 mL) and dioxane (15.0 mL) was added 3-bromo-2,4-dimethoxypyridine (961.2 mg, 4.48 mmol), Pd(AMPHOS)2C12 (260.1 mg, 0.37 mmol) and K3PO4 (2.4 g, 11.24 mmol). The resulting mixture was stirred at 80° C. for 12 h under N2. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 6-chloro-3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (830.0 mg, 54%) as a red oil. LCMS (ESI, m/z): [M+H]+=420.1.
To a solution of 6-chloro-3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (830.0 mg, 1.98 mmol) in dioxane (10.0 mL) was added diphenylmethanimine (1.0 g, 5.94 mmol), Brettphos Pd G3 (194.5 mg, 0.20 mmol), Brettphos (216.4 mg, 0.40 mmol) and Cs2CO3 (1936.4 mg, 5.94 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/2, v/v) to afford N-(3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-1,1-diphenylmethanimine (1.0 g, 89%) as a light red oil. LCMS (ESI, m/z): [M+H]+=565.3.
To a solution of N-(3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-1,1-diphenylmethanimine (1.0 g, 0.47 mmol) in DCM (10.0 mL) was added FA (5.0 mL) at room temperature. The mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O. The pH value of the mixture was adjusted to 7 with NaHCO3 solution. The mixture was extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (530.0 mg, 75%) as a red oil. LCMS (ESI, m/z): [M+H]+=401.2.
To a solution of 3-(2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (200.0 mg, 0.50 mmol) in pyridine/DCM (10.0 mL/5.0 mL) was added phenyl chloroformate (78.18 mg, 0.499 mmol) at 0° C. under N2. The mixture was stirred at 0° C. for 1 h. The resulting mixture was concentrated under vacuum. To the above mixture was added a solution of 2-(4-ethylpiperazin-1-yl)ethan-1-amine (235.7 mg, 1.49 mmol) in pyridine (10.0 mL). The resulting mixture was stirred at 60° C. for 2 h under N2. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (9/1, v/v) to afford 3-[3-(2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-ethylpiperazin-1-yl)ethyl]urea (90.0 mg, 31%) as an off-white solid. LCMS (ESI, m/z): [M+H]+=584.3.
To a solution of 3-[3-(2,4-dimethoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-ethylpiperazin-1-yl)ethyl]urea (70.0 mg, 0.12 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 10% B to 40% B in 9 min; 254 nm) to afford 3-[3-(2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-ethylpiperazin-1-yl)ethyl]urea (Compound 194) (6.8 mg, 13%) as a white solid. LCMS (ESI, m/z): [M+H]+=454.3. 1H NMR (400 MHz, DMSO-d6): δ 11.40 (s, 1H), 9.16 (s, 1H), 8.35 (s, 1H), 8.05 (d, J=5.6 Hz, 1H), 7.52 (d, J=8.8 Hz, 1H), 7.25 (d, J=2.4 Hz, 1H), 7.03 (d, J=8.4 Hz, 1H), 6.87 (d, J=6.0 Hz, 1H), 3.80 (s, 3H), 3.79 (s, 3H), 2.52-2.44 (m, 5H), 2.36-2.26 (m, 5H), 0.99-0.96 (m, 3H).
To a solution of 3-bromo-2,4-dimethoxypyridine (380.0 mg, 1.74 mmol) in 1,4-dioxane/H2O (10.0 mL/2.0 mL) was added (6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)boronic acid (683.1 mg, 2.09 mmol), K3PO4 (1109.8 mg, 5.23 mmol) and (AMPhosPdCl2)2 (123.4 mg, 0.17 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (60/40, v/v) to afford 6-chloro-3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (220.0 mg, 30%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=420.1.
To a solution of 6-chloro-3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (180.0 mg, 0.43 mmol) in 1,4-dioxane (10.0 mL) was added NH2-Boc (65.3 mg, 0.56 mmol), Cs2CO3 (418.9 mg, 1.29 mmol), XPhos (40.9 mg, 0.09 mmol) and Pd(OAc)2 (9.6 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (50/50, v/v) to afford tert-butyl (3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)carbamate (200.0 mg, 93%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=501.2.
To a solution of tert-butyl (3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)carbamate (180.0 mg, 0.45 mmol) in CH2Cl2 (10.0 mL) was added FA (15.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (140.0 mg, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=401.2.
To a solution of 3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (100.0 mg, 0.25 mmol) in CH2Cl2 (10.0 mL) was added phenyl carbonochloridate (117.3 mg, 0.75 mmol) and pyridine (80.0 mg, 1.00 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in pyridine (10.0 mL). Then N1, N1-dimethylpropane-1,3-diamine (30.6 mg, 0.30 mmol) was added to the mixture at room temperature. The resulting mixture was stirred at 60° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (86/14, v/v) to afford 1-(3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(3-(dimethylamino)propyl)urea (72.0 mg, 55%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=529.3.
To a solution of 1-(3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(3-(dimethylamino)propyl)urea (120.0 mg, 0.23 mmol) in CH2Cl2 (4.0 mL) was added TFA (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (4.0 mL) was added NH3·H2O (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 47% B in 9 min; 254 nm) to afford 1-(3-(2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(3-(dimethylamino)propyl)urea (Compound 195) (14.6 mg, 16%) as a white solid. LCMS (ESI, m/z): [M+H]+=399.3. H NMR (400 MHz, DMSO-d6): δ 11.45 (s, 1H), 9.13 (s, 1H), 8.57 (s, 1H), 8.05 (d, J=6.0 Hz, 1H), 7.53 (d, J=8.4 Hz, 1H), 7.26 (d, J=2.4 Hz, 1H), 6.95 (d, J=8.4 Hz, 1H), 6.87 (d, J=6.0 Hz, 1H), 3.80 (s, 3H), 3.79 (s, 3H), 3.33-3.21 (m, 1H), 3.25-3.23 (m, 1H), 2.34-2.23 (m, 2H), 2.17 (s, 6H), 1.68-1.62 (m, 2H).
To a solution of 3-chloro-5-methoxyaniline (2.0 g, 12.69 mmol) in DMF (10.0 mL) was added NIS (2.6 g, 11.42 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 0.5 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (85/15, v/v) to afford 3-chloro-4-iodo-5-methoxyaniline (1.6 g, 44%) as an off-white solid. LCMS (ESI, m/z): [M+H]+=283.9.
To a solution of 3-chloro-4-iodo-5-methoxyaniline (1.6 g, 5.64 mmol) in acetone (24.0 mL) was added benzoyl isothiocyanate (921.6 mg, 5.64 mmol) at 0° C. under N2. The resulting mixture was stirred at 60° C. for 0.5 h. After the reaction was completed, the reaction mixture was filtered. The solid was collected and dried to afford 1-benzoyl-3-(3-chloro-4-iodo-5-methoxyphenyl)thiourea (2.0 g, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=446.9.
To a solution of 1-benzoyl-3-(3-chloro-4-iodo-5-methoxyphenyl)thiourea (2.0 g, 4.38 mmol) in MeOH/H2O (10.0/10.0 mL) was added NaOH (193.3 mg, 4.83 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the mixture was filtered. The solid was collected and dried to afford 3-chloro-4-iodo-5-methoxyphenylthiourea (1.4 g, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=342.9.
To a solution of 3-chloro-4-iodo-5-methoxyphenylthiourea (700.0 mg, 2.04 mmol) in CHCl3 (8.0 mL) was added Br2 (359.2 mg, 2.25 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the reaction was quenched with saturated NaHSO3 solution at 0° C. The resulting mixture was extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 7-chloro-6-iodo-5-methoxy-1,3-benzothiazol-2-amine (560.0 mg, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=340.9.
To a solution of 7-chloro-6-iodo-5-methoxy-1,3-benzothiazol-2-amine (560.0 mg, 1.72 mmol) in THF (10.0 mL) was added t-BuONO (265.3 mg, 2.57 mmol) and DMSO (10.7 mg, 0.14 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (85/15, v/v) to afford 7-chloro-6-iodo-5-methoxy-1,3-benzothiazole (250.0 mg, 44%) as a white solid. LCMS (ESI, m/z): [M+H]+=325.9.
To a solution of 7-chloro-6-iodo-5-methoxy-1,3-benzothiazole (220.0 mg, 0.68 mmol) in 1,4-dioxane/H2O (8.0 mL/1.6 mL) was added 6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-ylboronic acid (264.9 mg, 0.81 mmol), K2CO3 (280.2 mg, 2.03 mmol) and Pd(dppf)Cl2 (55.0 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford 7-chloro-6-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-5-methoxy-1,3-benzothiazole (218.0 mg, 67%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=480.1.
To a solution of 7-chloro-6-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-5-methoxy-1,3-benzothiazole (188.0 mg, 0.39 mmol) in 1,4-dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (201.7 mg, 1.96 mmol), K2CO3 (162.2 mg, 1.17 mmol), BrettPhos (42.0 mg, 0.08 mmol) and Pd2(dba)3 (35.8 mg, 0.04 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum CH2Cl2/CH3OH (95/5, v/v) to afford (1S,2S)—N-[3-(7-chloro-5-methoxy-1,3-benzothiazol-6-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (140.0 mg, 52%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=547.1.
To a solution of (1S,2S)—N-[3-(7-chloro-5-methoxy-1,3-benzothiazol-6-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (130.0 mg, 0.24 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with CH3CN/H2O (70/30, v/v) to afford (1S,2S)—N-[3-(7-chloro-5-methoxy-1,3-benzothiazol-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (Compound 196) (12.3 mg, 11%) as a white solid. LCMS (ESI, m/z): [M+H]+=417.1. 1H NMR (400 MHz, DMSO-d6): δ 11.77 (s, 1H), 10.68 (s, 1H), 9.45 (s, 1H), 7.87 (d, J=8.4 Hz, 1H), 7.79 (s 1H), 7.61 (d, J=8.8 Hz, 1H), 7.49 (d, J=2.0 Hz, 1H), 5.03-4.82 (m, 1H), 3.82 (s, 3H), 2.30-2.19 (m, 1H), 1.70-1.62 (m, 1H), 1.23-1.11 (m, 1H).
To a solution of 5-fluoro-4-methylpyridin-2-amine (600.0 mg, 4.76 mmol) in H2SO4 (12.0 mL)/H2O (24.0 mL) was added dropwise a solution of NaNO2 (1641.0 mg, 23.78 mmol) in H2O (12.0 mL) at 0° C. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 5-fluoro-4-methylpyridin-2-ol (200.0 mg, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=128.0.
To a solution of 5-fluoro-4-methylpyridin-2-ol (820.0 mg, 6.45 mmol) in ACN (30.0 mL) was added NIS (1.6 g, 7.10 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/4, v/v) to afford 5-fluoro-3-iodo-4-methylpyridin-2-ol (1.6 g, 98%) as a brown solid. LCMS (ESI, m/z): [M+H]+=253.9.
To a solution of 5-fluoro-3-iodo-4-methylpyridin-2-ol (900.0 mg, 3.56 mmol) in CHCl3 (20.0 mL) was added Ag2CO3 (3923.4 mg, 14.23 mmol) and CH3I (2019.6 mg, 14.23 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 4 h. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 5-fluoro-3-iodo-2-methoxy-4-methylpyridine (130.0 mg, 13%) as a white solid. LCMS (ESI, m/z): [M+H]+=268.0
To a solution of 5-fluoro-3-iodo-2-methoxy-4-methylpyridine (380.0 mg, 1.42 mmol) in 1,4-dioxane/H2O (10.0/2.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (581.7 mg, 1.42 mmol), XPhos (135.7 mg, 0.29 mmol), K3PO4 (906.2 mg, 4.27 mmol) and XPhos Pd G3 (120.5 mg, 0.14 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-fluoro-2-methoxy-4-methylpyridine (320.0 mg, 53%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=422.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-5-fluoro-2-methoxy-4-methylpyridine (320.0 mg, 0.76 mmol) in 1,4-dioxane (10.0 mL) were added (1S,2S)-2-fluorocyclopropane-1-carboxamide (390.9 mg, 3.79 mmol), BrettPhos (81.4 mg, 0.15 mmol), Cs2CO3 (741.3 mg, 2.28 mmol) and BrettPhos Pd G3 (68.7 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 4 h under N2. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/3, v/v) to afford (1S,2S)-2-fluoro-N-[3-(5-fluoro-2-methoxy-4-methylpyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (180.0 mg, 48%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=489.2.
To a solution of (1S,2S)-2-fluoro-N-[3-(5-fluoro-2-methoxy-4-methylpyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (180.0 mg, 0.37 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (3.0 mL) was added NH3·H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 56% B in 12 min; 254 nm) to afford (1S,2S)-2-fluoro-N-[3-(5-fluoro-2-methoxy-4-methylpyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropane-1-carboxamide (Compound 197) (27.0 mg, 20%) as a white solid. LCMS (ESI, m/z): [M+H]+=359.1. 1H NMR (300 MHz, DMSO-d6): δ 11.74 (s, 1H), 10.67 (s, 1H), 8.09 (s, 1H), 7.87 (d, J=8.1 Hz, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.46 (s, 1H), 5.05-4.81 (m, 1H), 3.77 (s, 3H), 2.25-2.16 (m, 4H), 1.70-1.61 (m, 1H), 1.19-1.12 (m, 1H).
To a solution of 3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (500.0 mg, 1.35 mmol) in CH2Cl2 (5.0 mL) was added pyridine (428.1 mg, 5.41 mmol) and phenyl chloroformate (254.2 mg, 1.62 mmol) at 0° C. The mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum. To the above residue was added pyridine (5.0 mL) and tert-butyl 3-aminoazetidine-1-carboxylate (699.1 mg, 4.06 mmol). The resulting mixture was stirred at 60° C. for 2 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (97/3, v/v) to afford tert-butyl 3-([[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)azetidine-1-carboxylate (600.0 mg, 78%) as a brown oil. LCMS (ESI, m/z): [M+H]+=568.3.
To a solution of tert-butyl 3-([[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]carbamoyl]amino)azetidine-1-carboxylate (600.0 mg, 1.06 mmo) in CH2Cl2 (2.0 mL) was added HCOOH (4.0 mL). The resulting mixture was stirred at room temperature for 6 h. After the reaction was completed, The pH value of the mixture was adjusted to 7 with aq·NaHCO3. The resulting mixture was extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum to afford 3-(azetidin-3-yl)-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (480.0 mg, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=468.2.
To a solution of 3-(azetidin-3-yl)-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (480.0 mg, crude) in CH2Cl2 (5.0 mL) was added 2-(dimethylamino)acetaldehyde hydrochloride (380.5 mg, 3.08 mmol) and NaBH3CN (193.5 mg, 3.08 mmol) at room temperature. The mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with CH2Cl2/MeOH (97/3, v/v) to afford 3-[1-[2-(dimethylamino)ethyl]azetidin-3-yl]-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (150.0 mg, 27%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=539.3.
To a solution of 3-[1-[2-(dimethylamino)ethyl]azetidin-3-yl]-1-[3-(2-methoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]urea (150.0 mg, 0.28 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL). The mixture was stirred at room temperature for 3 h. The mixture was concentrated under vacuum. To the above residue was added ACN (3.0 mL) and NH3·H2O (3.0 mL). The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the resulting mixture was concentrated under vacuum. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 48% B in 8 min; 254 nm) to afford 3-[1-[2-(dimethylamino)ethyl]azetidin-3-yl]-1-[3-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]urea (Compound 198) (11.4 mg, 10%) as a white solid. LCMS (ESI, m/z): [M+H]+=409.3. 1H NMR (300 MHz, DMSO-d6): δ 11.62 (s, 1H), 9.24 (s, 1H), 8.98-8.95 (m, 1H), 7.92 (d, J=8.4 Hz, 1H), 7.53-7.50 (m, 2H), 7.30-7.24 (m, 1H), 7.11 (d, J=7.5 Hz, 1H), 7.04-6.99 (m, 2H), 4.37-4.30 (m, 1H), 3.81 (s, 3H), 3.68-3.57 (m, 2H), 3.00-2.95 (m, 2H), 2.63-2.55 (m, 2H), 2.36-2.27 (m, 2H), 2.22 (s, 6H).
To a solution of 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (350.0 mg, 1.07 mmol) in 1,4-dioxane (2.0 mL) was added 6-bromo-5-methoxy-1,3-benzothiazole (260.8 mg, 1.07 mmol), K3PO4 (680.2 mg, 3.21 mmol), Pd(AMPHOS)2C12 (90.8 mg, 0.13 mmol) and H2O (0.5 mL) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (60/40, v/v) to afford 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)-5-methoxybenzo[d]thiazole (195.0 mg, 41%) as a white solid. LCMS (ESI, m/z): [M+H]+=447.1.
To a solution of 6-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)-5-methoxybenzo[d]thiazole (150.0 mg, 0.34 mmol) in 1,4-dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (173.0 mg, 1.68 mmol), K2CO3 (139.1 mg, 1.01 mmol) BrettPhos (36.0 mg, 0.07 mmol) and BrettPhos Pd G3 (30.4 mg, 0.03 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (70/30, v/v) to afford (1S,2S)-2-fluoro-N-(3-(5-methoxybenzo[d]thiazol-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)cyclopropane-1-carboxamide (66.0 mg, 38%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=514.2.
To a solution of (1S,2S)-2-fluoro-N-(3-(5-methoxybenzo[d]thiazol-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridin-6-yl)cyclopropane-1-carboxamide (46.0 mg, 0.09 mmol) in DCM (1.0 mL) was added TFA (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in ACN (1.0 mL) was added NH3·H2O (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 ml/min; Gradient: 28% B to 35% B in 8 min; 254 nm) to afford (1S,2S)-2-fluoro-N-(3-(5-methoxybenzo[d]thiazol-6-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl)cyclopropane-1-carboxamide (Compound 199) (15.3 mg, 44%) as a white solid. LCMS (ESI, m/z): [M+H]+=384.1. 1H NMR (300 MHz, DMSO-d6): δ 13.57 (s, 1H), 10.99 (s, 1H), 9.42 (s, 1H), 8.35 (s, 1H), 8.14 (d, J=8.8 Hz, 1H), 8.01 (d, J=9.2 Hz, 1H), 7.84 (s, 1H), 5.05-4.87 (m, 1H), 3.94 (s, 3H), 2.29-2.26 (m, 1H), 1.72-1.64 (m, 1H), 1.22-1.15 (m, 1H).
To a solution of 3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (200.0 mg, 0.50 mmol) in CH2Cl2 (5.0 mL) was added pyridine (80.0 mg, 2.50 mmol) and phenyl chloroformate (156.7 mg, 1.0 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. The resulting mixture was concentrated under reduced pressure. To the above mixture in pyridine (5.0 mL) was added tert-butyl 3-(aminomethyl)azetidine-1-carboxylate (372.9 mg, 2.00 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (90/10, v/v) to afford tert-butyl 3-((3-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)ureido)methyl)azetidine-1-carboxylate (150.0 mg, 49%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=612.3.
To a solution of tert-butyl 3-((3-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)ureido)methyl)azetidine-1-carboxylate (100.0 mg, 0.16 mmol) in DCM (2.0 mL) was added FA (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the pH value of the mixture was adjusted to 7 with aq·NaHCO3 and then extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 1-(azetidin-3-ylmethyl)-3-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)urea (170.0 mg, crude) as a brown solid. LCMS (ESI, m/z): [M+H]+=512.3.
To a solution of 1-(azetidin-3-ylmethyl)-3-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)urea (170.0 mg, 0.33 mmol) in MeOH (3.0 mL) was added HCHO (50.0 mg, 0.67 mmol) and NaBH4 (75.4 mg, 1.99 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 4 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 1-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-((1-methylazetidin-3-yl)methyl)urea (190.0 mg, crude) as a brown solid. LCMS (ESI, m/z): [M+H]+=526.3.
To a solution of 1-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-((1-methylazetidin-3-yl)methyl)urea (200.0 mg, 0.38 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 31% B to 40% B in 8 min; 254 nm) to afford 1-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-((1-methylazetidin-3-yl)methyl)urea (Compound 200) (5.2 mg, 3%) as a white solid. LCMS (ESI, m/z): [M+H]+=396.2. 1H NMR (400 MHz, DMSO-d6): δ 11.37 (s, 1H), 9.14 (s, 1H), 8.82 (s, 1H), 7.46 (d, J=8.4 Hz, 1H), 7.28-7.24 (m, 1H), 7.16 (d, J=2.4 Hz, 1H), 6.88 (d, J=8.8 Hz, 1H), 6.74 (d, J=8.8 Hz, 2H), 3.68 (s, 6H), 3.40-3.37 (m, 3H), 3.30-3.24 (m, 2H), 2.92-2.85 (m, 2H), 2.18 (s, 3H).
To a solution of 2-chloro-5-fluoropyridin-4-ol (2.0 g, 13.55 mmol) in CH3OH (10.0 mL) was added NBS (2.9 g, 16.26 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (92/8, v/v) to afford 3-bromo-2-chloro-5-fluoropyridin-4-ol (3.0 g, 97%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=225.9.
To a solution of 3-bromo-2-chloro-5-fluoropyridin-4-ol (1.5 g, 6.62 mmol) in DMF (5.0 mL) was added K2CO3 (1.8 g, 13.25 mmol) and CH3I (1.1 g, 7.95 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (60/40, v/v) to afford 3-bromo-2-chloro-5-fluoro-4-methoxypyridine (600.0 mg, 37%) as a white solid. LCMS (ESI, m/z): [M+H]+=239.9.
To a solution of 3-bromo-2-chloro-5-fluoro-4-methoxypyridine (600.0 mg, 2.50 mmol) in EtOH (5.0 mL) was added EtONa (203.6 mg, 3.00 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (65/35, v/v) to afford 3-bromo-2-ethoxy-5-fluoro-4-methoxypyridine (500.0 mg, 80%) as a white solid. LCMS (ESI, m/z): [M+H]+=250.0.
To a solution of 3-bromo-2-ethoxy-5-fluoro-4-methoxypyridine (400.0 mg, 1.60 mmol) in 1,4-dioxane/H2O (5.0/1.0 mL) was added 6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-ylboronic acid (627.0 mg, 1.92 mmol), K3PO4 (679.2 mg, 3.20 mmol), 3-tert-butyl-4-(2,6-dimethoxyphenyl)-2,3-dihydro[d]-[1,3]-oxaphosphole (CAS 1246888-90-3) (52.8 mg, 0.16 mmol) and Pd2(dba)3 CHCl3 (80.0 mg, 0.08 mmol) at room temperature under N2. The resulting mixture was stirred at 90° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (0/100, v/v) to afford 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-ethoxy-5-fluoro-4-methoxypyridine (300.0 mg, 55%) as a white solid. LCMS (ESI, m/z): [M+H]+=452.1.
To a solution of 3-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-2-ethoxy-5-fluoro-4-methoxypyridine (300.0 mg, 0.66 mmol) in 1,4-dioxane (5.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (342.1 mg, 3.32 mmol), Cs2CO3 (648.7 mg, 1.99 mmol), BrettPhos (71.1 mg, 0.13 mmol) and BrettPhos Pd G3 (60.6 mg, 0.07 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (0/100, v/v) to afford (1S,2S)—N-[3-(2-ethoxy-5-fluoro-4-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (160.0 mg, 46%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=519.2.
To a solution of (1S,2S)—N-[3-(2-ethoxy-5-fluoro-4-methoxypyridin-3-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (160.0 mg, 0.31 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (3.0 mL) was added NH3·H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 27% B in 9 min, 254 nm) to afford (1S,2S)—N-[3-(2-ethoxy-5-fluoro-4-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (Compound 201) (37.5 mg, 31%) as a white solid. LCMS (ESI, m/z): [M+H]+=389.1. 1H NMR (400 MHz, DMSO-d6): δ 11.58 (s, 1H), 10.63 (s, 1H), 8.06 (d, J=7.2 Hz, 1H), 7.84 (d, J=8.8 Hz, 1H), 7.75 (d, J=8.4 Hz, 1H), 7.59 (d, J=2.4 Hz, 1H), 5.00-4.83 (m, 1H), 3.63-3.57 (m, 5H), 2.27-2.18 (m, 1H), 1.67-1.61 (m, 1H), 1.17-1.12 (m, 1H), 0.96-0.90 (m, 3H).
To a solution of 3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-amine (180.0 mg, 0.45 mmol) in DCM (10.0 mL) were added pyridine (142.5 mg, 1.80 mmol) and phenyl chloroformate (141.1 mg, 0.90 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the above mixture were added 1-[1-[(dimethylamino)methyl]cyclopropyl]methanamine dihydrochloride (453.1 mg, 2.25 mmol) and pyridine (10.0 mL) at room temperature. The resulting mixture was stirred at 60° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/methanol (6/1, v/v) to afford 1-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-3-([1-[(dimethylamino)methyl]cyclopropyl]methyl)urea (174.0 mg, 69%) as a light brown solid. LCMS (ESI, m/z): [M+H]+=554.3.
To a solution of 1-[3-(2,6-dimethoxyphenyl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-6-yl]-3-([1-[(dimethylamino)methyl]cyclopropyl]methyl)urea (174.0 mg, 0.31 mmol) in CH2Cl2 (8.0 mL) was added TFA (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (8.0 mL) was added NH3·H2O (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 47% B in 10 min; 254/220 nm) to afford 1-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-([1-[(dimethylamino)methyl]cyclopropyl]methyl)urea (Compound 202) (30.6 mg, 22%) as a white solid. LCMS (ESI, m/z): [M+H]+=424.2. 1H NMR (400 MHz, DMSO-d6): δ 11.26 (d, J=1.6 Hz, 1H), 9.11 (s, 1H), 8.46 (s, 1H), 7.46 (d, J=8.8 Hz, 1H), 7.28-7.24 (m, 1H), 7.15 (d, J=2.4 Hz, 1H), 6.96 (d, J=8.4 Hz, 1H), 6.74 (d, J=8.4 Hz, 2H), 3.68 (s, 6H), 3.33-3.24 (m, 2H), 2.20-2.18 (m, 8H), 0.54-0.51 (m, 2H), 0.29-0.26 (m, 2H).
To a solution of 6-chloro-3-(4,6-dimethoxy-2-methylpyrimidin-5-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (200.0 mg, 0.46 mmol) in THE (10.0 mL) was added LiHMDS (0.9 mL, 1 mol/L), Xphos (43.8 mg, 0.09 mmol) and Pd2(dba)3 (42.1 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred at 60° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 3-(4,6-dimethoxy-2-methylpyrimidin-5-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (160.0 mg, 83%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=416.2.
To a solution of 3-(4,6-dimethoxy-2-methylpyrimidin-5-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (130.0 mg, 0.31 mmol) in CH2Cl2 (5.0 mL) was added phenyl carbonochloridate (146.9 mg, 0.94 mmol) and pyridine (100.2 mg, 1.25 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in pyridine (5.0 mL) was added 2-(4-ethylpiperazin-1-yl)ethan-1-amine (59.0 mg, 0.38 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 1-(3-(4,6-dimethoxy-2-methylpyrimidin-5-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(2-(4-ethylpiperazin-1-yl)ethyl)urea (80.0 mg, 42%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=599.3.
To a solution of 1-(3-(4,6-dimethoxy-2-methylpyrimidin-5-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(2-(4-ethylpiperazin-1-yl)ethyl)urea (70.0 mg, 0.12 mmol) in DCM (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in ACN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 31% to 41% in 8 min; 254 nm) to afford 1-(3-(4,6-dimethoxy-2-methylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(2-(4-ethylpiperazin-1-yl)ethyl)urea (Compound 203) (12.1 mg, 22%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=469.4. 1H NMR (300 MHz, CDCl3): δ 10.14 (s, 1H), 9.73 (s, 1H), 7.64 (d, J=8.4 Hz, 1H), 7.36 (d, J=2.4 Hz, 1H), 6.43 (d, J=8.4 Hz, 1H), 3.99 (s, 6H), 3.66-3.60 (m, 2H), 2.90-2.83 (m, 6H), 2.66-2.52 (m, 9H), 1.21-1.16 (m, 3H).
To a solution of 3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (375.0 mg, 0.88 mmol) in DCM (15.0 mL) was added pyridine (278.1 mg, 3.52 mmol) and phenyl chloroformate (275.3 mg, 1.76 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added (3-amino-2-fluoropropyl)dimethylamine (950.7 mg, 7.91 mmol) and pyridine (15.0 mL) at room temperature. The resulting mixture was stirred at 70° C. for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (11/1, v/v) to afford 1-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea (420.0 mg, 83%) as a light yellow oil. LCMS (ESI, m/z): [M+H]+=573.3.
To a solution of 1-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea (270.0 mg, 0.47 mmol) in CH2Cl2 (10.0 mL) was added TFA (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (10.0 mL) was added NH3·H2O (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with acetonitrile/water (1/1, v/v) to afford 1-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea (95.0 mg, 45%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=443.2.
The racemic 1-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea (95.0 mg, 0.22 mmol) was separated by Prep-Chiral—HPLC with the following conditions (Column: CHIRAL ART Amylose-SA, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 50% B to 50% B in 21 min; Wave Length: 220/254 nm) to afford 1-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 1 (31.4 mg, 66%) as a white solid and 1-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 2 (35.4 mg, 74%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 204 and 205 in Table 1.
1-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 1: Retention Time 1=6.66 min; LCMS (ESI, m/z): [M+H]+=443.1. 1H NMR (400 MHz, DMSO-d6): δ 11.49 (s, 1H), 9.23 (s, 1H), 8.62 (s, 1H), 8.07 (d, J=5.6 Hz, 1H), 7.51 (d, J=8.4 Hz, 1H), 7.23 (d, J=2.0 Hz, 1H), 7.13 (d, J=6.0 Hz, 1H), 7.00 (d, J=8.8 Hz, 1H), 4.81-4.68 (m, 1H), 3.94-3.91 (m, 1H), 3.81 (s, 3H), 3.65-3.52 (m, 1H), 3.45-3.35 (m, 1H), 2.56-2.51 (m, 2H), 2.22 (s, 6H), 0.81-0.76 (m, 2H), 0.63-0.58 (m, 2H).
1-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 2: Retention Time 2=14.70 min; LCMS (ESI, m/z): [M+H]+=443.1. 1H NMR (400 MHz, DMSO-d6): δ 11.49 (s, 1H), 9.23 (s, 1H), 8.63 (s, 1H), 8.06 (d, J=5.6 Hz, 1H), 7.51 (d, J=8.8 Hz, 1H), 7.23 (d, J=2.0 Hz, 1H), 7.13 (d, J=6.0 Hz, 1H), 7.00 (d, J=8.4 Hz, 1H), 4.82-4.69 (m, 1H), 3.93-3.90 (m, 1H), 3.81 (s, 3H), 3.63-3.52 (m, 1H), 3.45-3.34 (m, 1H), 2.57-2.51 (m, 2H), 2.23 (s, 6H), 0.81-0.76 (m, 2H), 0.65-0.63 (m, 2H).
To a solution of 3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (300.0 mg, 0.66 mmol) in CH2Cl2 (4.0 mL) was added pyridine (207.4 mg, 2.62 mmol) and phenyl chloroformate (123.2 mg, 0.79 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure. To the residue in pyridine (6.0 mL) was added (3-amino-2-fluoropropyl)dimethylamine (787.8 mg, 6.56 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 1 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum CH2Cl2/CH3OH (90/10, v/v) to afford 1-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl)-3-[3-(dimethylamino)-2-fluoropropyl]urea (250.0 mg, 63%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=604.2.
To a solution of 1-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl)-3-[3-(dimethylamino)-2-fluoropropyl]urea (250.0 mg, 0.41 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with ACN/H2O (52/48, v/v) and then separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRAL ART Amylose-SA, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 50% B to 50% B in 10.5 min; Wave Length: 220/254 nm) to afford 1-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 1 (30.9 mg, 31%) as a white solid and 1-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 2 (40.4 mg, 41%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 206 and 207 in Table 1.
1-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 1: Retention Time 1=5.876 min; LCMS (ESI, m/z): [M+H]+=474.2. 1H NMR (400 MHz, DMSO-d6): δ 11.62 (s, 1H), 9.56 (s, 1H), 9.27 (s, 1H), 8.62 (s, 1H), 7.65 (d, J=8.8 Hz, 1H), 7.37 (d, J=2.0 Hz, 1H), 7.05 (d, J=8.8 Hz, 1H), 4.82-4.69 (m, 1H), 3.91-3.86 (m, 6H), 3.65-3.53 (m, 1H), 3.46-3.33 (m, 1H), 2.56-2.51 (m, 2H), 2.21 (s, 6H).
1-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-[3-(dimethylamino)-2-fluoropropyl]urea Enantiomer 2: Retention Time 2=9.106 min; LCMS (ESI, m/z): [M+H]+=474.2. 1H NMR (400 MHz, DMSO-d6): δ 11.62 (s, 1H), 9.56 (s, 1H), 9.27 (s, 1H), 8.62 (s, 1H), 7.65 (d, J=8.4 Hz, 1H), 7.37 (s, 1H), 7.05 (d, J=8.8 Hz, 1H), 4.82-4.69 (m, 1H), 3.91 and 3.89 (s, total 6H), 3.62-3.54 (m, 1H), 3.44-3.33 (m, 1H), 2.67-2.46 (m, 2H), 2.21 (s, 6H).
To a solution of 3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (1.0 g, 2.34 mmol) in DMF (20.0 mL) was added trans-2-(methoxycarbonyl)cyclopropane-1-carboxylic acid (404.5 mg, 2.81 mmol), DIEA (1.5 g, 11.70 mmol) and HATU (1.3 g, 3.51 mmol) at 0 C under N2. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (86/14, v/v) to afford methyl (trans)-2-{[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}cyclopropane-1-carboxylate (510.0 mg, 39%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=554.2.
To a solution of methyl (trans)-2-{[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}cyclopropane-1-carboxylate (460.0 mg, 0.83 mmol) in THF/CH3OH (12.0/3.0 mL) was added NaBH4 (3.1 g, 83.10 mmol) at room temperature. The resulting mixture was stirred at 30° C. for 16 h. After the reaction was completed, the reaction mixture was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (57/43, v/v) to afford trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (265.0 mg, 61%) as a white solid. LCMS (ESI, m/z): [M+H]+=526.2.
To a solution of trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (250.0 mg, 0.48 mmol) in CH2Cl2 (10.0 mL) was added Dess-Martin (302.6 mg, 0.71 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (250.0 mg, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=524.2.
To a solution of trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (250.0 mg, crude) in CH2Cl2 (10.0 mL) were added dimethylamine hydrochloride (116.8 mg, 1.43 mmol) and NaBH3CN (90.0 mg, 1.43 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum CH2Cl2/CH3OH (10/01, v/v) to afford trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide (150.0 mg, 57%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=553.3.
To a solution of trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide (130.0 mg, 0.24 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 m; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 20% B to 30% B in 8 min, 30% B; Wave Length: 254 nm) to afford trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide (50.0 mg, 50%) as a white solid. LCMS (ESI, m/z): [M+H]+=423.2.
The product trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide (50.0 mg, 0.12 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRAL ART Cellulose-SC, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: MeOH: EtOH=1: 1—HPLC; Flow rate: 20 mL/min; Gradient: 30% B to 30% B in 12.5 min; Wave Length: 220/254 nm) to afford N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 1 (6.6 mg, 26%) and N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 2 (5.4 mg, 22%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 208 and 209 in Table 1.
N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1=9.02 min; LCMS (ESI, m/z): [M+H]+=423.4. 1H NMR (400 MHz, DMSO-d6): δ 11.62 (s, 1H), 10.54 (s, 1H), 8.49 (s, 1H), 7.85 (d, J=8.8 Hz, 1H), 7.65 (d, J=8.8 Hz, 1H), 7.44 (d, J=2.4 Hz, 1H), 4.38-4.35 (m, 1H), 3.92 (s, 3H), 2.31-2.27 (m, 1H), 2.19-2.14 (m, 7H), 1.90-1.87 (m, 1H), 1.35-1.33 (m, 1H), 1.07-1.03 (m, 1H), 0.76-0.70 (m, 3H), 0.69-0.62 (m, 2H).
N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2=11.39 min; LCMS (ESI, m/z): [M+H]+=423.4. 1H NMR (400 MHz, DMSO-d6): δ 11.62 (s, 1H), 10.54 (s, 1H), 8.49 (s, 1H), 7.86 (d, J=8.8 Hz, 1H), 7.65 (d, J=8.8 Hz, 1H), 7.44 (d, J=2.4 Hz, 1H), 4.38-4.35 (m, 1H), 3.92 (s, 3H), 2.31-2.26 (m, 1H), 2.19-2.13 (m, 7H), 1.91-1.88 (m, 1H), 1.35-1.33 (m, 1H), 1.07-1.03 (m, 1H), 0.78-0.62 (m, 5H). $$
To a solution of 3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (170.0 mg, 0.40 mmol) in pyridine/DCM (3.0 mL/25.0 mL) was added phenyl chloroformate (124.8 mg, 0.80 mmol) at 0° C. The mixture was stirred at 0° C. for 1 h. The resulting mixture was concentrated under vacuum. To the above residue was added a solution of 2-(4-methylpiperazin-1-yl)ethanamine (285.4 mg, 2.00 mmol) in pyridine (10.0 mL). The resulting mixture was stirred at 60° C. for 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (10/1, v/v) to afford 3-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (110.0 mg, 46%) as a red oil. LCMS (ESI, m/z): [M+H]+=596.3.
To a solution of 3-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-1-[2-(4-methylpiperazin-1-yl)ethyl]urea (100.0 mg, 0.168 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 5% B to 25% B in 8 min; 254 nm) to afford 1-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(2-(4-methylpiperazin-1-yl)ethyl)urea formate (Compound 210) (55.7 mg, 71%) as a white solid. LCMS (ESI, m/z): [M+H]+=466.4. 1H NMR (400 MHz, Methanol-d4): δ 8.27 (s, 1H), 8.04 (d, J=5.6 Hz, 1H), 7.59 (d, J=8.8 Hz, 1H), 7.28 (s, 1H), 7.17 (d, J=6.0 Hz, 1H), 6.70 (d, J=8.4 Hz, 1H), 3.93-3.89 (m, 4H), 3.59-3.56 (m, 2H), 3.21-3.17 (m, 4H), 2.92 (s, 3H), 2.83-2.80 (m, 6H), 0.85-0.81 (m, 2H), 0.70-0.65 (m, 2H).
To a solution of trans-N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (400.0 mg, crude) in CH2Cl2 (20.0 mL) was added 1-ethylpiperazine (276.5 mg, 2.42 mmol) and NaBH3CN (152.1 mg, 2.42 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction mixture was quenched with MeOH. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (8/1, v/v) to afford trans-N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (180.0 mg, 38%) as a white solid. LCMS (ESI, m/z): [M+H]+=594.3.
To a solution of trans-N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (150.0 mg, 0.26 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with CH3OH/H2O (7/1, v/v) to afford trans-N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (65.7 mg, 55%) as a white solid. LCMS (ESI, m/z): [M+H]+=464.3.
The product trans-N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (65.7 mg, 0.14 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IG, 2×25 cm, 5 m; Mobile Phase A:Hex:DCM=3:1 (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 20% B to 20% B in 14 min; Wave Length: 220/254 nm) to afford N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 1 (10.7 mg, 16%) as a white solid and N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 2 (8.5 mg, 13%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 211 and 212 in Table 1.
N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1=7.55 min; LCMS (ESI, m/z): [M+H]+=464.3. 1H NMR (400 MHz, DMSO-d6): δ 11.40 (d, J=1.6 Hz, 1H), 10.52 (s, 1H), 7.81 (d, J=8.4 Hz, 1H), 7.52 (d, J=8.4 Hz, 1H), 7.29-7.25 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 3.69 (s, 6H), 2.46-2.21 (m, 10H), 1.91-1.88 (m, 1H), 1.36-1.31 (m, 1H), 1.05-0.97 (m, 4H), 0.78-0.65 (m, 1H).
N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2=11.05 min; LCMS (ESI, m/z): [M+H]+=464.3. 1H NMR (400 MHz, DMSO-d6): δ 11.40 (d, J=1.6 Hz, 1H), 10.52 (s, 1H), 7.81 (d, J=8.4 Hz, 1H), 7.52 (d, J=8.4 Hz, 1H), 7.30-7.25 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 3.69 (s, 6H), 2.46-2.21 (m, 10H), 1.90-1.88 (m, 1H), 1.36-1.31 (m, 1H), 1.05-0.97 (m, 4H), 0.78-0.65 (m, 1H).
To a solution of 5-bromo-4,6-dichloropyrimidine (30.0 g, 131.64 mmol) in MeOH/H2O (200.0/200.0 mL) was added MeONa (6.4 g, 118.47 mmol) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 5-bromo-4-chloro-6-methoxypyrimidine (31.0 g, crude) as a white solid. LCMS (ESI, m/z): [M+H]+=222.9.
To a solution of 5-bromo-4-chloro-6-methoxypyrimidine (31.0 g, crude) in DMF (120.0 mL) was added Cs2CO3 (57.2 g, 175.59 mmol) and cyclopropanol (8.5 g, 146.31 mmol) at room temperature under N2. The reaction mixture was stirred at 80° C. for 6 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (91/9, v/v) to afford 5-bromo-4-cyclopropoxy-6-methoxypyrimidine (9.0 g, 12%) as a white solid. LCMS (ESI, m/z): [M+H]+=245.0.
To a solution of 5-bromo-4-cyclopropoxy-6-methoxypyrimidine (8.0 g, 24.48 mmol) in 1.4-dioxane/H2O (100.0/20.0 mL) was added 6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-ylboronic acid (12.8 g, 39.17 mmol), K2CO3 (13.6 g, 97.92 mmol) and Pd(dppf)Cl2 (5.3 g, 6.51 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (86/14, v/v) to afford 5-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-4-cyclopropoxy-6-methoxypyrimidine (5.8 g, 39%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=447.2.
To a solution of 5-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-4-cyclopropoxy-6-methoxypyrimidine (5.0 g, 11.18 mmol) in 1,4-dioxane (120.0 mL) was added NH2Boc (6.5 g, 55.82 mmol), Cs2CO3 (27.8 g, 33.54 mmol), XPhos (1.7 g, 2.23 mmol) and Pd(OAc)2 (251.2 mg, 1.11 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (92/8, v/v) to afford tert-butyl N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamate (5.0 g, 70%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=528.3.
A solution of tert-butyl N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamate (5.0 g, 7.56 mmol) in FA/CH2Cl2 (20.0/20.0 mL) was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was adjusted pH to 7 with aq. NaHCO3 and then extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (3.0 g, crude) as a yellow oil. LCMS (ESI, m/z): [M+H]+=428.2.
To a solution of 3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (2.0 g, crude) in DMF (40.0 mL) was added trans-2-(methoxycarbonyl)cyclopropane-1-carboxylic acid (2.1 g, 4.54 mmol), DIEA (2.4 g, 18.94 mmol) and HATU (2.2 g, 5.68 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (88/12, v/v) to afford methyl (trans)-2-{[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}cyclopropane-1-carboxylate (1.0 g, 47%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=554.2.
To a solution of methyl (trans)-2-{[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}cyclopropane-1-carboxylate (1.0 g, 1.80 mmol) in THF/MeOH (5.0/5.0 mL) was added NaBH4 (2.1 g, 54.18 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed. the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated in vacuo. The residue was purified by reverse phase flash column chromatography with petroleum CH3CN/H2O (60/40, v/v) to afford trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (560.0 mg, 70%) as a white solid. LCMS (ESI, m/z): [M+H]+=526.2.
To a solution of trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (560.0 mg, 1.23 mmol) in CH2Cl2 (8.0 mL) was added Dess-Martin (1.2 g, 3.00 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum to afford trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (400.0 mg, crude) as a yellow oil. LCMS (ESI, m/z): [M+H]+=524.2.
To a solution of trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (400.0 mg, crude) in CH2Cl2 (5.0 mL) was added 1-methylpiperazine (400.0 mg, 3.99 mmol) and NaBH3CN (150.0 mg, 2.40 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under vacuum. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (60/40, v/v) to afford trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (200.0 mg, 62%) as a white solid. LCMS (ESI, m/z): [M+H]+=608.3.
To a solution of trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (200.0 mg, 0.33 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with CH3CN/H2O (7/4, v/v) to afford trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (110.0 mg, 46%) as a white solid. LCMS (ESI, m/z): [M+H]+=478.2.
The product trans-N-(3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (110.0 mg, 0.20 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRAL ART Cellulose-SC, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 30% B to 30% B in 15 min; Wave Length: 220/254 nm) to afford N-(3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide Enantiomer 1 (8.4 mg, 10%) as a white solid and N-(3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide Enantiomer 2 (9.0 mg, 14%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 213 and 214 in Table 1.
N-(3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1=11.427 min; LCMS (ESI, m/z): [M+H]+=478.4. 1H NMR (400 MHz, CD3OD): δ 8.41 (s, 1H), 7.78-7.70 (m, 2H), 7.43 (s, 1H), 4.42-4.39 (m, 1H), 4.00 (s, 3H), 2.81-2.53 (m, 7H), 2.43-2.23 (m, 5H), 1.83-1.78 (m, 1H), 1.64-1.54 (m, 1H), 1.43-1.21 (m, 2H), 0.98-0.52 (m, 5H).
N-(3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2=13.695 min; LCMS (ESI, m/z): [M+H]+=478.4. 1H NMR (400 MHz, CD3OD): δ 8.41 (s, 1H), 7.76-7.68 (m, 2H), 7.41 (s, 1H), 4.40-4.37 (m, 1H), 3.98 (s, 3H), 2.80-2.51 (m, 7H), 2.39-2.29 (m, 5H), 1.81-1.76 (m, 1H), 1.62-1.52 (m, 1H), 1.33-1.24 (m, 2H), 0.90-0.66 (m, 5H).
To a solution of 4-cyclopropoxy-3-iodo-2-methoxypyridine (300.0 mg, 1.03 mmol) in dioxane/H2O (10.0/2.0 mL) was added 6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridin-3-ylboronic acid (337.7 mg, 1.03 mmol), K3PO4 (656.3 mg, 3.09 mmol), (AMPhosPdCl2)2 (145.9 mg, 0.20 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with ACN/H2O (2/1, v/v) to afford 3-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridin-3-yl)-4-cyclopropoxy-2-methoxypyridine (80.0 mg, 16%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=447.2.
To a solution of 3-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridin-3-yl)-4-cyclopropoxy-2-methoxypyridine (110.0 mg, 0.25 mmol) in 1,4-dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (126.8 mg, 1.23 mmol), BrettPhos (26.4 mg, 0.05 mmol), Cs2CO3 (240.5 mg, 0.74 mmol) and BrettPhos Pd G3 (22.3 mg, 0.03 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 16 h. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford (1S,2S)—N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (80.0 mg, 63%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=514.2.
To a solution of (1S,2S)—N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrazolo[3,4-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (70.0 mg, 0.14 mmol) in DCM (4.0 mL) was added TFA (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. Then NH3·H2O (4.0 mL) and ACN (4.0 mL) were added to the residue at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column (Xselect CSH OBD Column 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 30% B to 40% B in 8 min; 254 nm) to afford (1S,2S)—N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrazolo[3,4-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (Compound 215) (12.5 mg, 24%) as a white solid. LCMS (ESI, m/z): [M+H]+=384.3. 1H NMR (400 MHz, DMSO-d6): δ 13.41 (s, 1H), 10.95 (s, 1H), 8.22 (d, J=5.6 Hz, 1H), 7.94 (d, J=8.8 Hz, 1H), 7.81 (d, J=8.8 Hz, 1H), 7.19 (d, J=5.6 Hz, 1H), 5.06-4.85 (m, 1H), 4.02-3.93 (m, 1H), 3.79 (s, 3H), 2.28-2.25 (m, 1H), 1.71-1.64 (m, 1H), 1.22-1.20 (m, 1H), 0.83-0.74 (m, 2H), 0.59-0.51 (m, 2H).
To a solution of 3-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-4-cyclopropoxy-2-methoxypyridine (950.0 mg, 2.13 mmol) in THE (30.0 mL) was added LiHMDS (6.4 mL, 2 mol/L), XPhos (203.1 mg, 0.43 mmol) and Pd2(dba)3 (122.5 mg, 0.13 mmol) at room temperature under N2. The resulting mixture was stirred at 60° C. for 1 h. After the reaction was completed, the reaction mixture was quenched by the addition of sat. NH4Cl (aq.). The reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford 3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (690.0 mg, 76%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=427.2.
To a solution of 3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (640.0 mg, 1.50 mmol) in DMF (20.0 mL) was added DIEA (969.5 mg, 7.50 mmol), HATU (855.7 mg, 2.25 mmol) and trans-2-(methoxycarbonyl)cyclopropane-1-carboxylic acid (259.5 mg, 1.80 mmol) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the mixture was purified by reverse phase flash chromatography with ACN/H2O (2/1, v/v) to afford methyl (trans)-2-{[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}cyclopropane-1-carboxylate (720.0 mg, 87%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=553.2.
To a solution of methyl (trans)-2-{[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}cyclopropane-1-carboxylate (700.0 mg, 1.27 mmol) in THE (6.0 mL)/CH3OH (4.0 mL) was added NaBH4 (958.3 mg, 25.34 mmol) at room temperature. The resulting mixture was stirred at 40° C. for 3 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with ACN/H2O (2/1, v/v) to afford trans-N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (600.0 mg, 90%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=525.2.
To a solution of trans-N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (200.0 mg, 0.38 mmol) in DCM (10.0 mL) was added Dess-Martin (242.5 mg, 0.57 mmol) at 0° C. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction mixture was quenched by the addition of sat. NaHCO3 (aq.). The reaction mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (190.0 mg, crude) as a green oil. LCMS (ESI, m/z): [M+H]+=523.2.
To a solution of trans-N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (190.0 mg, crude) in DCM (6.0 mL) was added 1-methylpiperazine (109.2 mg, 1.09 mmol) and NaBH3CN (68.5 mg, 1.09 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) to afford trans-N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (100.0 mg, 45%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=607.3.
To a solution of trans-N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (280.0 mg, 0.46 mmol) in CH2Cl2 (10.0 mL) was added TFA (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (10.0 mL) was added NH3·H2O (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with ACN/H2O (2/1, v/v) to afford trans-N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (100.0 mg, 45%) as a white solid. LCMS (ESI, m/z): [M+H]+=477.3.
The product trans-N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (100.0 mg, 0.18 mmol) was separated by Prep-Chiral-HPLC with the following conditions Column: Column (CHIRAL ART Cellulose-SC, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: MeOH: EtOH=1:1—HPLC; Flow rate: 20 mL/min; Gradient: 40% B to 40% B in 20 min; Wave Length: 220/254 nm) to afford N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 1 (26.5 mg, 29%) as a white solid and N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 2 (20.4 mg, 22%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 216 and 217 in Table 1.
N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1=18.75 min; LCMS (ESI, m/z): [M+H]+=477.4. 1H NMR (400 MHz, DMSO-d6): δ 11.50 (s, 1H), 10.52 (s, 1H), 8.07 (d, J=6.0 Hz, 1H), 7.83 (d, J=8.8 Hz, 1H), 7.56 (d, J=8.8 Hz, 1H), 7.34 (d, J=2.4 Hz, 1H), 7.14 (d, J=6.0 Hz, 1H), 3.94-3.91 (m, 1H), 3.82 (s, 3H), 2.44-2.22 (m, 7H), 2.14 (s, 3H), 1.90-1.88 (m, 1H), 1.34-1.31 (m, 1H), 1.06-1.03 (m, 1H), 0.79-0.76 (m, 2H), 0.71-0.64 (m, 3H).
N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 2 Retention Time 2=23.58 min; LCMS (ESI, m/z): [M+H]+=477.4. 1H NMR (400 MHz, DMSO-d6): δ 11.50 (s, 1H), 10.52 (s, 1H), 8.07 (d, J=6.0 Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 7.56 (d, J=8.8 Hz, 1H), 7.34 (d, J=2.4 Hz, 1H), 7.14 (d, J=6.0 Hz, 1H), 3.94-3.91 (m, 1H), 3.82 (s, 3H), 2.45-2.22 (m, 9H), 2.14 (s, 3H), 1.90-1.88 (m, 1H), 1.34-1.31 (m, 1H), 1.06-1.03 (m, 1H), 0.79-0.76 (m, 2H), 0.71-0.64 (m, 3H).
To a solution of trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-formylcyclopropane-1-carboxamide (400.0 mg, crude) in CH2Cl2 (5.0 mL) was added dimethylamine hydrochloride (103.5 mg, 1.27 mmol) and NaBH3CN (144.3 mg, 3.30 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with acetonitrile/water (16/84, v/v) to afford trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (168.0 mg, 40%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=552.3.
To a solution of trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (158.0 mg, 0.29 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with acetonitrile/water (65/35, v/v) to afford trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (51.0 mg, 41%) as a white solid. LCMS (ESI, m/z): [M+H]+=422.4.
The product trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (100.0 mg, 0.23 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRAL ART Cellulose-SC, 2×25 cm, 5 um; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: MeOH:EtOH=1:1—HPLC; Flow rate: 20 mL/min; Gradient: 30% B to 30% B in 14 min; 254 nm) to afford N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide Enantiomer 1 (11.1 mg, 9%) as a white solid and N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide Enantiomer 2 (19.1 mg, 16%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 218 and 219 in Table 1.
N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1=9.93 min; LCMS (ESI, m/z): [M+H]+=422.4. 1H NMR (400 MHz, DMSO-d6): δ 11.50 (s, 1H), 10.52 (s, 1H), 8.07 (d, J=5.2 Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 7.56 (d, J=8.8 Hz, 1H), 7.35 (s, 1H), 7.14 (d, J=4.4 Hz, 1H), 3.97-3.91 (m, 1H), 3.82 (s, 3H), 2.41-2.22 (m, 8H), 1.93-1.87 (m, 1H), 1.35-1.24 (m, 1H), 1.10-1.02 (m, 1H), 0.88-0.62 (m, 5H).
N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2=12.38 min; LCMS (ESI, m/z): [M+H]+=422.3. 1H NMR (400 MHz, DMSO-d6): δ 11.50 (s, 1H), 10.52 (s, 1H), 8.07 (d, J=5.6 Hz, 1H), 7.82 (d, J=8.4 Hz, 1H), 7.55 (d, J=8.8 Hz, 1H), 7.34 (d, J=2.4 Hz, 1H), 7.13 (d, J=5.6 Hz, 1H), 3.93-3.90 (m, 1H), 3.81 (s, 3H), 2.33-2.16 (m, 8H), 1.91-1.88 (m, 1H), 1.34-1.30 (m, 1H), 1.06-1.02 (m, 1H), 0.84-0.60 (m, 5H).
To a solution of 3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (1.9 g, 4.09 mmol) in DMF (20.0 mL) was DIEA (2.6 g, 20.4 mmol), trans-2-(methoxycarbonyl)cyclopropane-1-carboxylic acid (588.9 mg, 4.09 mmol) and HATU (1.9 g, 4.90 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 2 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (66/34, v/v) to afford methyl (trans)-2-((3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)carbamoyl)cyclopropane-1-carboxylate (1.6 g, 67%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=584.2.
To a solution of methyl (trans)-2-((3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)carbamoyl)cyclopropane-1-carboxylate (1.5 g, 2.57 mmol) in THF/CH3OH (20.0/5.0 mL) was NaBH4 (2.9 g, 77.2 mmol) at room temperature. The resulting mixture was stirred at 40° C. for 16 h. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (60/40, v/v) to afford trans-N-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(hydroxymethyl)cyclopropane-1-carboxamide (800.0 mg, 50%) as a white solid. LCMS (ESI, m/z): [M+H]+=556.2.
To a solution of trans-N-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(hydroxymethyl)cyclopropane-1-carboxamide (660.0 mg, 1.19 mmol) in CH2Cl2 (15.0 mL) was Dess-Martin (755.6 mg, 1.78 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with aq. NaHCO3, brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-formylcyclopropane-1-carboxamide (850.0 mg, crude) as a brown solid. LCMS (ESI, m/z): [M+H]+=554.2.
To a solution of trans-N-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-formylcyclopropane-1-carboxamide (850.0 mg, 1.54 mmol) in CH2Cl2 (20.0 mL) was added 1-methylpiperazine (307.5 mg, 3.07 mmol) and NaBH3CN (192.9 mg, 3.07 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (10/1, v/v) to afford trans-N-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (170.0 mg, 17%) as a brown solid. LCMS (ESI, m/z): [M+H]+=638.3.
To a solution of trans-N-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (170.0 mg, 0.27 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (10/1, v/v) to afford trans-N-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (100.0 mg, 70%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=508.2.
The product trans-N-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (100.0 mg, 0.20 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRAL ART Cellulose-SC, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: MeOH: EtOH=1:1—HPLC; Flow rate: 20 mL/min; Gradient: 50% B to 50% B in 14.5 min; Wave Length: 220/254 nm) to afford N-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide Enantiomer 1 (20.7 mg, 41%) as a white solid and N-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide Enantiomer 2 (19.7 mg, 39%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 220 and 221 in Table 1.
N-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1: 10.25 min; LCMS (ESI, m/z): [M+H]+=508.4. 1H NMR (400 MHz, DMSO-d6): δ 11.65 (s, 1H), 10.57 (s, 1H), 9.57 (d, J=5.6 Hz, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.69 (d, J=8.4 Hz, 1H), 7.47 (s, 1H), 3.91 (s, 6H), 2.44-2.11 (m, 9H), 2.10-2.01 (m, 4H), 1.91-1.89 (m, 1H), 1.37-1.33 (m, 1H), 1.11-1.00 (m, 1H), 0.77-0.64 (m, 1H).
N-(3-(5,7-dimethoxythiazolo[4,5-b]pyridin-6-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide Enantiomer 2 Retention Time 2:12.63 min; LCMS (ESI, m/z): [M+H]+=508.4. 1H NMR (400 MHz, DMSO-d6): δ 11.65 (s, 1H), 10.57 (s, 1H), 9.56 (s, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.68 (d, J=8.8 Hz, 1H), 7.47 (s, 1H), 3.91 (s, 6H), 2.44-2.20 (m, 10H), 2.14 (s, 3H), 1.91-1.89 (m, 1H), 1.35-1.32 (m, 1H), 1.11-1.00 (m, 1H), 0.77-0.64 (m, 1H).
To a solution of 6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-ylboronic acid (2.0 g, 6.12 mmol) in 1,4-dioxane/H2O (40.0 mL/8.0 mL) was added 1-fluoro-2-iodo-3-methoxybenzene (1.5 g, 6.12 mmol), K2CO3 (1.7 g, 12.25 mmol) and Pd(dppf)Cl2 (448.0 mg, 0.61 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford 6-chloro-3-(2-fluoro-6-methoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridine (2.3 g, 92%) as a brown oil. LCMS (ESI, m/z): [M+H]+=407.1.
To a solution of 6-chloro-3-(2-fluoro-6-methoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridine (1.0 g, 2.46 mmol) in THE (10.0 mL) was added XPhos (351.4 mg, 0.74 mmol), Pd2(dba)3 (225.0 mg, 0.25 mmol) and LiHMDS (7.4 mL, 1 mol/L) at room temperature under N2. The resulting mixture was stirred at 60° C. for 2 h. After the reaction was completed, the reaction mixture was quenched by the addition of sat. NH4Cl (aq.) at room temperature. The resulting mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (4/1, v/v) to afford 3-(2-fluoro-6-methoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (790.0 mg, 82%) as a brown oil. LCMS (ESI, m/z): [M+H]+=388.2.
To a solution of 3-(2-fluoro-6-methoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (770.0 mg, 1.99 mmol) in DMF (20.0 mL) was added DIEA (1.2 g, 9.94 mmol), trans-2-(methoxycarbonyl)cyclopropane-1-carboxylic acid (429.6 mg, 2.98 mmol) and HATU (1.4 g, 3.58 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with acetonitrile/water (4/1, v/v) to afford methyl trans-2-{[3-(2-fluoro-6-methoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}cyclopropane-1-carboxylate (760.0 mg, 74%) as a light brown oil. LCMS (ESI, m/z): [M+H]+=514.2.
To a solution of methyl trans-2-{[3-(2-fluoro-6-methoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}cyclopropane-1-carboxylate (750.0 mg, 1.46 mmol) in THF/MeOH (12.0 mL/8.0 mL) was added NaBH4 (1.7 g, 43.8 mmol) at room temperature. The resulting mixture was stirred at 40° C. for 0.5 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with acetonitrile/water (7/3, v/v) to afford trans-N-[3-(2-fluoro-6-methoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (365.0 mg, 51%) as a white solid. LCMS (ESI, m/z): [M+H]+=486.2.
To a solution of trans-N-[3-(2-fluoro-6-methoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (340.0 mg, 0.70 mmol) in CH2Cl2 (10.0 mL) was added Dess-Martin (445.4 mg, 1.05 mmol) at 0° C. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed. the resulting mixture was diluted with water and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-[3-(2-fluoro-6-methoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (330.0 mg, crude) as a green oil. LCMS (ESI, m/z): [M+H]+=484.2.
To a solution of trans-N-[3-(2-fluoro-6-methoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (650.0 mg, crude) in DCM (10.0 mL) was added 1-methylpiperazine (403.9 mg, 4.03 mmol) and NaBH3CN (253.4 mg, 4.03 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (9/1, v/v) to afford trans-N-[3-(2-fluoro-6-methoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (150.0 mg, 19%) as a light brown oil. LCMS (ESI, m/z): [M+H]+=568.3.
To a solution of trans-N-[3-(2-fluoro-6-methoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (140.0 mg, 0.25 mmol) in CH2Cl2 (6.0 mL) was added TFA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (6.0 mL) was added NH3·H2O (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with acetonitrile/water (1/4, v/v) to afford trans-N-[3-(2-fluoro-6-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (80.0 mg, 74%) as a light yellow solid. LCMS (ESI, m/z): [M+H]+=438.2.
The product trans-N-[3-(2-fluoro-6-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (80.0 mg, 0.18 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IE, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: MeOH: EtOH=1: 1—HPLC; Flow rate: 16 mL/min; Gradient: 60% B to 60% B in 29 min; Wave Length: 220/254 nm) to afford N-[3-(2-fluoro-6-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 1 (13.2 mg, 15%) as a white solid and N-[3-(2-fluoro-6-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 2 (12.3 mg, 15%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 222 and 223 in Table 1.
N-[3-(2-fluoro-6-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1=15.57 min; LCMS (ESI, m/z): [M+H]+=438.4. 1H NMR (400 MHz, DMSO-d6): δ 11.63 (s, 1H), 10.55 (s, 1H), 7.87 (d, J=8.4 Hz, 1H), 7.66 (d, J=8.4 Hz, 1H), 7.44 (s, 1H), 7.36-7.30 (m, 1H), 6.98-6.89 (m, 2H), 3.78 (s, 3H), 2.45-2.22 (m, 8H), 2.14 (s, 3H), 1.95-1.89 (m, 1H), 1.36-1.32 (m, 1H), 1.06-1.04 (m, 1H), 0.74-0.66 (m, 1H).
N-[3-(2-fluoro-6-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-methylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2=22.25 min; LCMS (ESI, m/z): [M+H]+=438.4. 1H NMR (400 MHz, DMSO-d6): δ 11.63 (s, 1H), 10.55 (s, 1H), 7.87 (d, J=8.4 Hz, 1H), 7.66 (d, J=8.8 Hz, 1H), 7.44 (s, 1H), 7.36-7.30 (m, 1H), 6.98-6.89 (m, 2H), 3.78 (s, 3H), 2.38-2.22 (m, 8H), 2.14 (s, 3H), 1.94-1.89 (m, 1H), 1.38-1.32 (m, 1H), 1.06-1.01 (m, 1H), 0.72-0.66 (m, 1H).
To a solution of 3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (900.0 mg, 2.52 mmol) in THE (10.0 mL) was added trans-ethyl-2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)cyclopropane-1-carboxylate (818.7 mg, 3.38 mmol) and AlMe3 (1.5 mL, 2 mol/L) at room temperature under N2. The resulting mixture was stirred at 80° C. for 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (90/10, v/v) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)cyclopropane-1-carboxamide (1.3 g, 87%) as a brown solid. LCMS (ESI, m/z): [M+H]+=596.3.
To a solution of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)cyclopropane-1-carboxamide (1.1 g, 1.84 mmol) in CH3OH (10.0 mL) was added TsOH (158.9 mg, 0.92 mmol) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. the residue was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (88/12, v/v) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-hydroxyethyl)cyclopropane-1-carboxamide (930.0 mg, 88%) as a brown solid. LCMS (ESI, m/z): [M+H]+=512.3.
To a solution of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-hydroxyethyl)cyclopropane-1-carboxamide (790.0 mg, 1.54 mmol) in CH2Cl2 (10.0 mL) was added Dess-Martin (982.2 mg, 2.32 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-oxoethyl)cyclopropane-1-carboxamide (790.0 mg, crude) as a brown solid. LCMS (ESI, m/z): [M+H]+=510.2.
To a solution of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-oxoethyl)cyclopropane-1-carboxamide (790.0 mg, crude) in CH2Cl2 (10.0 mL) was added dimethylamine hydrochloride (376.2 mg, 4.61 mmol) and NaBH3CN (290.0 mg, 4.61 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (90/10, v/v) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide (750.0 mg, 81%) as a brown solid. LCMS (ESI, m/z): [M+H]+=539.3.
To a solution of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide (750.0 mg, 1.26 mmol) in CH2Cl2 (4.0 mL) was added TFA (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (4.0 mL) was added NH3·H2O (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 8 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (10/1, v/v) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide (200.0 mg, 27%) as a white solid. LCMS (ESI, m/z): [M+H]+=409.2.
The product trans-N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide (200.0 mg, 0.48 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IH, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: MeOH: EtOH=1: 1—HPLC; Flow rate: 20 mL/min; Gradient: 20% B to 20% B in 19 min; Wave Length: 220/254 nm) to afford N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide Enantiomer 1 (23.0 mg, 23%) as a white solid and N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide Enantiomer 2 (20.6 mg, 20%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 224 and 225 in Table 1.
N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1 (min): 10.06; LCMS (ESI, m/z): [M+H]+=409.2. 1H NMR (400 MHz, DMSO-d6): δ 11.38 (s, 1H), 10.43 (s, 1H), 7.80 (d, J=8.8 Hz, 1H), 7.52 (d, J=8.4 Hz, 1H), 7.29-7.25 (m, 2H), 6.80-6.74 (m, 2H), 3.69 (s, 6H), 2.36-2.29 (m, 2H), 2.20-2.13 (m, 6H), 1.85-1.82 (m, 1H), 1.50-1.36 (m, 2H), 1.24-1.21 (m, 1H), 1.02-0.98 (m, 1H), 0.70-0.66 (m, 1H).
N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2 (min): 14.10; LCMS (ESI, m/z): [M+H]+=409.2. 1H NMR (400 MHz, DMSO-d6): δ 11.38 (s, 1H), 10.43 (s, 1H), 7.80 (d, J=8.4 Hz, 1H), 7.52 (d, J=8.8 Hz, 1H), 7.29-7.25 (m, 2H), 6.75 (d, J=8.4 Hz, 2H), 3.69 (s, 6H), 2.33-2.28 (m, 2H), 2.13 (s, 6H), 1.85-1.82 (m, 1H), 1.49-1.35 (m, 2H), 1.24-1.21 (m, 1H), 1.02-0.98 (m, 1H), 0.70-0.66 (m, 1H).
To a solution of 3-(4,6-dimethoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (474.0 mg, 1.18 mmol) in DCM (10.0 mL) was added Pyridine (373.5 mg, 4.72 mmol) and phenyl chloroformate (369.7 mg, 2.36 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added Pyridine (10.0 mL) and (3-amino-2-fluoropropyl)dimethylamine (1418.6 mg, 11.80 mmol) at room temperature. The resulting mixture was stirred at 60° C. for 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (20/1, v/v) to afford 1-[3-(4,6-dimethoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea (440.0 mg, 68%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=548.3.
To a solution of 1-[3-(4,6-dimethoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-3-[3-(dimethylamino)-2-fluoropropyl]urea (410.0 mg, 0.75 mmol) in CH2Cl2 (8.0 mL) was added TFA (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added CH3CN (8.0 mL) and NH3·H2O (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography eluted with methanol/H2O (3/2, v/v) to afford 1-(3-(4,6-dimethoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(3-(dimethylamino)-2-fluoropropyl)urea (95.0 mg, 30%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=418.2.
The racemic 1-(3-(4,6-dimethoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(3-(dimethylamino)-2-fluoropropyl)urea (95.0 mg, 0.23 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRAL ART Amylose-SA, 2×25 cm, m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 30% B to 30% B in 22 min; Wave Length: 220/254 nm) to afford 1-(3-(4,6-dimethoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(3-(dimethylamino)-2-fluoropropyl)urea Enantiomer 1 (18.9 mg, 20%) as a white solid and 1-(3-(4,6-dimethoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(3-(dimethylamino)-2-fluoropropyl)urea Enantiomer 2 (11.5 mg, 18%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 226 and 227 in Table 1.
1-(3-(4,6-dimethoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(3-(dimethylamino)-2-fluoropropyl)urea Enantiomer 1: Retention Time 1=10.16 min; LCMS (ESI, m/z): [M+H]+=418.3. 1H NMR (400 MHz, DMSO-d6): δ 11.61 (s, 1H), 9.27 (s, 1H), 8.61 (s, 1H), 8.46 (s, 1H), 7.65 (s, 1H), 7.37 (s, 1H), 7.05 (s, 1H), 4.81-4.69 (m, 1H), 3.91 (s, 6H), 3.74-3.55 (m, 1H), 2.21 (s, 6H).
1-(3-(4,6-dimethoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3-(3-(dimethylamino)-2-fluoropropyl)urea Enantiomer 2: Retention Time 2=17.388 min; LCMS (ESI, m/z): [M+H]+=418.3. 1H NMR (400 MHz, DMSO-d6): δ 11.62 (s, 1H), 9.27 (s, 1H), 8.61 (s, 1H), 8.46 (s, 1H), 7.65 (d, J=7.6 Hz, 1H), 7.37 (s, 1H), 7.05 (d, J=7.6 Hz, 1H), 4.81-4.69 (m, 1H), 3.91 (s, 6H), 3.60-3.54 (m, 1H), 2.21 (s, 6H).
To a solution of NaH (16.3 g, 60% purity) in DMF (250.0 mL) was added dropwise a solution of (E)-but-2-ene-1,4-diol (50.0 g, 567.50 mmol) in DMF (250.0 mL) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h under N2. Then a solution of BnBr (97.1 g, 567.50 mmol) in DMF (100.0 mL) was added dropwise to the mixture at 0° C. under N2. The resulting mixture was stirred at 0° C. for additional 1 h under N2. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (75/25, v/v) to afford (E)-4-(benzyloxy)but-2-en-1-ol (70.0 g, 69%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=179.1.
To a solution of (E)-4-(benzyloxy)but-2-en-1-ol (50.0 g, 280.54 mmol) in CH2Cl2 (500.0 mL) was added imidazole (42.0 g, 617.17 mmol), DMAP (6.9 g, 56.11 mmol) and TIPSCl (108.2 g, 561.07 mmol) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (96/4, v/v) to afford (E)-((4-(benzyloxy)but-2-en-1-yl)oxy)triisopropylsilane (90.0 g, 95%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=335.2.
To a solution of (E)-((4-(benzyloxy)but-2-en-1-yl)oxy)triisopropylsilane (59.0 g, 132.88 mmol) in DCM (300.0 mL) and NaOH (300.0 mL, 50%) was added NBu4I (4.90 g, 13.29 mmol) and CHBr2F (50.3 g, 199.32 mmol) at room temperature. The resulting mixture was stirred at room temperature for 12 h. After the reaction was completed, the resulting mixture was quenched with H2O at 0° C. and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford (((1S/R,3S/R)-3-((benzyloxy)methyl)-2-bromo-2-fluorocyclopropyl)methoxy)triisopropylsilane (50.0 g, crude) as a colorless oil. LCMS (ESI, m/z): [M+H]+=445.1.
To a solution of (((1S/R,3S/R)-3-((benzyloxy)methyl)-2-bromo-2-fluorocyclopropyl)methoxy)triisopropylsilane (50.0 g, 112.24 mmol) in hexane (500.0 mL) was added Bu3SnH (35.9 g, 123.4 mmol) and AIBN (1.8 g, 11.22 mmol) at room temperature. The resulting mixture was stirred at 90° C. for 4 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (88/12, v/v) to afford (trans-(2-((benzyloxy)methyl)-3-fluorocyclopropyl)methoxy)triisopropylsilane (50.0 g, crude) as a colorless oil. LCMS (ESI, m/z): [M+H]+=367.2.
To a solution of (trans-(2-((benzyloxy)methyl)-3-fluorocyclopropyl)methoxy)triisopropylsilane (50.0 g, 136.39 mmol) in THE (500.0 mL) was added TBAF (42.8 g, 163.67 mmol) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (40/60, v/v) to afford trans-(2-((benzyloxy)methyl)-3(S)-fluorocyclopropyl)methanol (1.5 g, 10%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=211.1.
To a solution of (COCl)2 (1.8 g, 14.27 mmol) in CH2Cl2 (10.0 mL) was added dropwise a solution of DMSO (4.5 g, 57.08 mmol) in CH2Cl2 (10.0 mL) at −78° C. under N2. The resulting mixture was stirred at −78° C. for 0.5 h under N2. Then a solution of trans-(2-((benzyloxy)methyl)-3(S)-fluorocyclopropyl)methanol (1.5 g, 7.13 mmol) in CH2Cl2 (10.0 mL) was added dropwise to the mixture at −78° C. under N2. The resulting mixture was stirred at −78° C. for additional 1 h. After the reaction was completed, TEA (5.8 g, 57.08 mmol) was added dropwise to the mixture at −78° C. under N2. The resulting mixture was stirred at −78° C. for another 0.5 h under N2. The resulting mixture was quenched with aqueous NH4Cl and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (90/10, v/v) to afford trans-(2-((benzyloxy)methyl)-3(R)-fluorocyclopropyl)-1-carbaldehyde (410.0 mg, 25%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=209.1.
To a mixture of trans-(2-((benzyloxy)methyl)-3(R)-fluorocyclopropyl)-1-carbaldehyde (360.0 mg, 1.73 mmol) and NH2SO3H (1.7 g, 17.29 mmol) in t-BuOH/H2O (1.0/8.0 mL) was added NaClO2 (1.6 g, 17.29 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 1.5 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (50/50, v/v) to afford trans-(2-((benzyloxy)methyl)-3(R)-fluorocyclopropyl)-1-carboxylic acid (80.0 mg, 20%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=225.1.
To a solution of 3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (231.7 mg, 0.58 mmol) in DMF (6.0 mL) were added trans-(2-((benzyloxy)methyl)-3(R)-fluorocyclopropyl)-1-carboxylic acid (130.0 mg, 0.58 mmol), DIEA (224.8 mg, 1.74 mmol) and HATU (264.5 mg, 0.70 mmol) at 0° C. under N2. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (90/10, v/v) to afford trans-2-((benzyloxy)methyl)-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(R)-fluorocyclopropane-1-carboxamide (270.0 mg, 77%) as a white solid. LCMS (ESI, m/z): [M+H]+=606.3.
To a solution of trans-2-((benzyloxy)methyl)-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(R)-fluorocyclopropane-1-carboxamide (230.0 mg, 0.38 mmol) in CH3OH (5.0 mL) was Pd(OH)2/C (106.1 mg, dry) at room temperature under N2. The resulting mixture was stirred at room temperature for 4 h under H2. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse flash column chromatography with CH3CN/H2O (28/72, v/v) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(R)-fluoro-2-(hydroxymethyl)cyclopropane-1-carboxamide (200.0 mg, 97%) as a white solid. LCMS (ESI, m/z): [M+H]+=516.2.
To a solution of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(R)-fluoro-2-(hydroxymethyl)cyclopropane-1-carboxamide (200.0 mg, 0.39 mmol) in CH2Cl2 (10.0 mL) was added Dess-Martin (246.8 mg, 0.58 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with aq. NaHCO3, brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(R)-fluoro-2-formylcyclopropane-1-carboxamide (200.0 mg, crude) as a brown solid. LCMS (ESI, m/z): [M+H]+=514.2.
To a solution of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(R)-fluoro-2-formylcyclopropane-1-carboxamide (200.0 mg, crude) in CH2Cl2 (10.0 mL) was added 1-methylpiperazine (117.0 mg, 1.17 mmol) and NaBH3CN (110.1 mg, 1.17 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (20/1, v/v) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(R)-fluoro-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (75.0 mg, 34%) as a brown solid. LCMS (ESI, m/z): [M+H]+=598.3.
To a solution of trans-N-(3-(2,6-dimethoxyphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(R)-fluoro-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (70.0 mg, 0.27 mmol) in CH2Cl2 (4.0 mL) was added TFA (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (4.0 mL) was added NH3·H2O (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for additional 8 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30×150 mm, 5 m; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 27% B to 37% B in 8 min; Wave Length: 254 nm) to afford trans-N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-3(R)-fluoro-2-((4-methylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (Compound 228) (3.6 mg, 7%) as a white solid. LCMS (ESI, m/z): [M+H]+=468.2. 1H NMR (400 MHz, DMSO-d6): δ 11.37 (s, 1H), 10.88 (s, 1H), 7.78 (d, J=8.0 Hz, 1H), 7.54 (d, J=8.4 Hz, 1H), 7.30-7.26 (m, 2H), 6.76 (d, J=8.4 Hz, 2H), 5.07-4.87 (m, 1H), 3.70 (s, 6H), 2.90-2.79 (m, 2H), 2.72-2.55 (m, 1H), 2.45-2.23 (m, 5H), 2.21-2.10 (m, 4H), 1.56-1.39 (m, 1H).
To a solution of trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-formylcyclopropane-1-carboxamide (250.0 mg, 0.47 mmol) in DCM (4.0 mL) was added NaBH3CN (90.1 mg, 1.43 mmol) and 1-ethylpiperazine (163.8 mg, 1.43 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (10/1, v/v) to afford trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((4-ethylpiperazin-1-yl)methyl)cyclopropane-1-carboxamide (75.0 mg, 25%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=621.4
To a solution of trans-N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (150.0 mg, 0.09 mmol) in DCM (2.0 mL) was added TFA (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added NH3·H2O (1.0 mL) and ACN (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for additional 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with ACN/H2O (2/1, v/v) to afford trans-N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (60.0 mg, 51%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=491.3
The trans-N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (60.0 mg) was separated by Prep-chiral HPLC with the following conditions (Column: CHIRAL ART Cellulose-SC, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 30% B to 30% B in 11 min; Wave Length: 220/254 nm) to afford N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 1 (11.8 mg, 39%) and N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 2 (17.4 mg, 56%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 229 and 230 in Table 1.
N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1=7.61 min; LCMS (ESI, m/z): [M+H]+=491.2. 1H NMR (400 MHz, DMSO-d6): δ 11.50 (s, 1H), 10.53 (s, 1H), 8.07 (d, J=6.0 Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 7.56 (d, J=8.8 Hz, 1H), 7.34 (d, J=2.0 Hz, 1H), 7.14 (d, J=2.0 Hz, 1H), 3.99-3.91 (m, 1H), 3.81 (s, 3H), 2.51-2.08 (m, 10H), 1.96-1.90 (m, 1H), 1.34-1.24 (m, 1H), 1.15-0.96 (m, 4H), 0.81-0.76 (m, 2H), 0.72-0.59 (m, 3H).
N-[3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 2 Retention Time 2=9.31 min; LCMS (ESI, m/z): [M+H]+=491.3. 1H NMR (400 MHz, DMSO-d6): δ 11.50 (s, 1H), 10.53 (s, 1H), 8.07 (d, J=5.6 Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 7.35 (d, J=2.0 Hz, 1H), 7.14 (d, J=5.6 Hz, 1H), 3.94-3.91 (m, 1H), 3.82 (s, 3H), 2.68-2.21 (m, 10H), 1.91-1.88 (m, 1H), 1.34-1.30 (m, 1H), 1.06-0.96 (m, 4H), 0.81-0.76 (m, 2H), 0.72-0.64 (m, 3H).
To a solution of 4,6-dimethoxypyridin-3-amine (2.0 g, 12.97 mmol) in MeCN (20.0 mL) was added t-BuONO (2.0 g, 19.46 mmol), CuCl2 (0.8 g, 6.48 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (9/1, v/v) to afford 5-chloro-2,4-dimethoxypyridine (900.0 mg, 39%) as a white solid. LCMS (ESI, m/z): [M+H]+=174.0
To a solution of 5-chloro-2,4-dimethoxypyridine (500.0 mg, 2.88 mmol) in AcOH (8.0 mL) was added NaOAc (236.2 mg, 2.88 mmol) and Br2 (690.4 mg, 4.32 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the pH value of the mixture was adjusted to 7 with aq·NaHCO3. The mixture was extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford 3-bromo-5-chloro-2,4-dimethoxypyridine (450.0 mg, 61%) as a white solid. LCMS (ESI, m/z): [M+H]+=251.9
To a solution of 3-bromo-5-chloro-2,4-dimethoxypyridine (400.0 mg, 1.67 mmol) in 1,4-dioxane/H2O (10.0/2.0 mL) was added Pd(PPh3)4 (387.6 mg, 0.33 mmol), 6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-ylboronic acid (657.5 mg, 2.01 mmol) and K2CO3 (695.4 mg, 5.03 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with ACN/H2O (6/1, v/v) to afford 5-chloro-3-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-2,4-dimethoxypyridine (250.0 mg, 32%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=454.1.
To a solution of 5-chloro-3-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-2,4-dimethoxypyridine (300.0 mg, 0.66 mmol) in dioxane (10.0 mL) was added (1S,2S)-2-fluorocyclopropane-1-carboxamide (0.3 g, 3.30 mmol), Pd2(dba)3 (0.1 g, 0.13 mmol), K2CO3 (0.3 g, 2.64 mmol) and BrettPhos (0.1 g, 0.26 mmol) at room temperature under N2. The resulting mixture was stirred at 100° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford (1S,2S)—N-[3-(5-chloro-2,4-dimethoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (150.0 mg, 43%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=521.2
To a solution of (1S,2S)—N-[3-(5-chloro-2,4-dimethoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (150.0 mg, 0.29 mmol) in DCM (2.0 mL) was added TFA (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the above residue was added NH3·H2O (2.0 mL) and ACN (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for additional 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Shield RP18 OBD Column, 19×250 mm, 10 m; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL/min; Gradient: 45% B to 60% B in 8 min, Wave Length: 254 nm) to afford (1S,2S)—N-[3-(5-chloro-2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-fluorocyclopropane-1-carboxamide (Compound 231) (10.6 mg, 9%) as a white solid. LCMS (ESI, m/z): [M+H]+=391.1. 1H NMR (400 MHz, DMSO-d6): δ 11.75 (s, 1H), 10.64 (s, 1H), 8.21 (s, 1H), 7.87 (d, J=8.4 Hz, 1H), 7.72 (d, J=8.4 Hz, 1H), 7.54 (s, 1H), 5.01-4.84 (m, 1H), 3.84 (s, 3H), 3.46 (s, 3H), 2.29-2.18 (m, 1H), 1.69-1.63 (m, 1H), 1.19-1.08 (m, 1H).
To a solution of 3-bromo-2,4-dimethoxypyridine (2.0 g, 9.17 mmol) in 1,4-dioxane (20.0 mL)/H2O (5.0 mL) was added 6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-ylboronic acid (3.6 g, 11.00 mmol), K2CO3 (3.8 g, 27.52 mmol) and Pd(dppf)Cl2 (671.1 mg, 0.92 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford 3-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-2,4-dimethoxypyridine (1.2 g, 31%) as a brown oil. LCMS (ESI, m/z): [M+H]+=420.1.
To a solution of 3-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-2,4-dimethoxypyridine (1.0 g, 2.38 mmol) in THF (40.0 mL) was added XPhos (454.0 mg, 0.95 mmol), Pd2(dba)3 (436.1 mg, 0.48 mmol) and LiHMDS (4.7 mL, 1 mol/L) at room temperature under N2. The resulting mixture was stirred at 60° C. for 1 h. After the reaction was completed, the reaction mixture was quenched with NH4Cl (aq) at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/1, v/v) to afford 3-(2,4-dimethoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (800.0 mg, 83%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=401.2.
To a solution of 3-(2,4-dimethoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (750.0 mg, 1.87 mmol) in DMF (20.0 mL) was added trans-2-(methoxycarbonyl)cyclopropane-1-carboxylic acid (323.2 mg, 2.24 mmol), DIEA (363.0 mg, 2.81 mmol) and HATU (1.1 g, 2.81 mmol) at 0° C. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford methyl (trans)-2-{[3-(2,4-dimethoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}cyclopropane-1-carboxylate (860.0 mg, 87%) as a brown oil. LCMS (ESI, m/z): [M+H]+=527.2.
To a solution of methyl (trans)-2-{[3-(2,4-dimethoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}cyclopropane-1-carboxylate (850.0 mg, 1.61 mmol) in THF (20.0 mL)/MeOH (30.0 mL) was added NaBH4 (3.1 g, 80.70 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction mixture was quenched with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/99, v/v) to afford trans-N-[3-(2,4-dimethoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (530.0 mg, 65%) as a brown solid. LCMS (ESI, m/z): [M+H]+=499.2.
To a solution of trans-N-[3-(2,4-dimethoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (500.0 mg, 1.00 mmol) in CH2Cl2 (20.0 mL) was added Dess-Martin (637.9 mg, 1.50 mmol) at 0° C. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-[3-(2,4-dimethoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (495.0 mg, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=497.2.
To a solution of trans-N-[3-(2,4-dimethoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (495.0 mg, 0.99 mmol) in DCM (20.0 mL) was added dimethylamine hydrochloride (245.0 mg, 3.02 mmol) and NaBH3CN (68.5 mg, 3.02 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction was quenched with water at room temperature. The resulting mixture was diluted with water and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford trans-N-(3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (170.0 mg, 32%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=526.3.
To a solution of trans-N-(3-(2,4-dimethoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (150.0 mg, 0.29 mmol) in CH2Cl2 (8.0 mL) was added TFA (8.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure. To the residue in ACN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (66/34, v/v) to afford trans-N-(3-(2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (80.0 mg, 70%) as a white solid. LCMS (ESI, m/z): [M+H]+=396.1.
The product trans-N-(3-(2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (80.0 mg, 0.20 mmol) was separated by Prep-Chiral—HPLC with the following conditions (Column: CHIRAL ART Cellulose-SC, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: MeOH: EtOH=1:1—HPLC; Flow rate: 20 mL/min; Gradient: 30% B to 30% B in 14.5 min; Wave Length: 220/254 nm) to afford N-[3-(2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 1 (8.7 mg, 21%) as a white solid and N-[3-(2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 2 (7.8 mg, 20%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 232 and 233 in Table 1.
N-[3-(2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1=10.43 min; LCMS (ESI, m/z): [M+H]+=396.1. 1H NMR (400 MHz, DMSO-d6): δ 11.51 (s, 1H), 10.51 (s, 1H), 8.07-8.02 (m, 1H), 7.83 (d, J=8.8 Hz, 1H), 7.60-7.55 (m, 1H), 7.37 (d, J=2.4 Hz, 1H), 6.89-6.82 (m, 1H), 3.98-3.80 (m, 6H), 2.35-2.27 (m, 1H), 2.19-2.15 (m, 7H), 1.96-1.89 (m, 1H), 1.36-1.28 (m, 1H), 1.11-1.04 (m, 1H), 0.72-0.68 (m, 1H).
N-[3-(2,4-dimethoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2=13.15 min; LCMS (ESI, m/z): [M+H]+=396.2. 1H NMR (400 MHz, DMSO-d6): δ 11.50 (s, 1H), 10.51 (s, 1H), 8.07 (d, J=5.6 Hz, 1H), 7.83 (d, J=8.8 Hz, 1H), 7.59 (d, J=8.8 Hz, 1H), 7.37 (d, J=2.4 Hz, 1H), 6.88 (d, J=6.0 Hz, 1H), 3.99-3.72 (m, 6H), 2.35-2.27 (m, 1H), 2.19-2.14 (m, 7H), 1.90-1.85 (m, 1H), 1.34-1.24 (m, 1H), 1.06-1.03 (m, 1H), 0.72-0.70 (m, 1H).
To a solution of trans-N-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl)-2-formylcyclopropane-1-carboxamide (400.0 mg, 0.72 mmol) in CH2Cl2 (4.0 mL) was added dimethylamine hydrochloride (176.7 mg, 2.17 mmol) and NaBH3CN (136.2 mg, 2.17 mmol) at room temperature. The resulting mixture was stirred at room temperature for 0.5 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography with CH2Cl2/CH3OH (12/1, v/v)) to afford trans-N-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl)-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide (300.0 mg, 75%) as a brown solid. LCMS (ESI, m/z): [M+H]+=583.2.
To a solution of trans-N-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl)-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide (300.0 mg, 0.52 mmol) in CH2Cl2 (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (3.0 mL) was added NH3·H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for additional 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with H2O/ACN (46/54, v/v) to afford trans-N-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide (60.0 mg, 25%) as a green solid. LCMS (ESI, m/z): [M+H]+=453.2.
The product of trans-N-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide (60.0 mg, 0.13 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRAL ART Cellulose-SC, 2×25 cm, 5 m; Mobile Phase A: Hex: DCM=3:1 (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 30% B to 30% B in 13 min; Wave Length: 220/254 nm) to afford N-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 1 (17.6 mg, 58%) as a green solid and N-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 2 (18.3 mg, 60%) as a green solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 234 and 235 in Table 1.
N-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1: 8.92 min; LCMS (ESI, m/z): [M+H]+=453.1. 1H NMR (400 MHz, DMSO-d6): δ 11.66 (d, J=1.6 Hz, 1H), 10.58 (s, 1H), 9.57 (s, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.69 (d, J=8.4 Hz, 1H), 7.48 (d, J=2.4 Hz, 1H), 3.91-3.87 (m, 6H), 2.33-2.15 (m, 8H), 1.91-1.85 (m, 1H), 1.36-1.32 (m, 1H), 1.07-1.01 (m, 1H), 0.72-0.65 (m, 1H).
N-(3-{5,7-dimethoxy-[1,3]thiazolo[4,5-b]pyridin-6-yl}-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2: 12.18 min; LCMS (ESI, m/z): [M+H]+=453.1. 1H NMR (400 MHz, DMSO-d6): δ 11.66 (d, J=1.6 Hz, 1H), 10.59 (s, 1H), 9.57 (s, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.69 (d, J=8.4 Hz, 1H), 7.48 (d, J=2.4 Hz, 1H), 3.91 (s, 6H), 2.39-2.20 (m, 8H), 1.93-1.88 (m, 1H), 1.38-1.33 (m, 1H), 1.08-1.05 (m, 1H), 0.76-0.65 (m, 1H).
To a solution of trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (170.0 mg, 0.33 mmol) in DCM (15.0 mL) was added 1-ethylpiperazine (111.2 mg, 0.98 mmol) and NaBH3CN (61.2 mg, 0.98 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (5/1, v/v) to afford trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (100.0 mg, 49%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=622.3.
To a solution of trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (300.0 mg, 0.48 mmol) in CH2Cl2 (6.0 mL) was added TFA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (6.0 mL) was added NH3·H2O (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with ACN/H2O (1/1, v/v) to afford trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (100.0 mg, 42%) as a white solid. LCMS (ESI, m/z): [M+H]+=492.3.
The trans-N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide (100.0 mg, 0.20 mmol) was separated by Prep-Chiral-HPLC with the following conditions Column: Column (CHIRAL ART Cellulose-SC, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 20% B to 20% B in 17 min; Wave Length: 220/254 nm) to afford N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 1 (21.5 mg, 42%) as a white solid and N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 2 (20.6 mg, 41%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 236 and 237 in Table 1.
N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1: 18.75 min; LCMS (ESI, m/z): [M+H]+=492.3. 1H NMR (400 MHz, DMSO-d6): δ 11.62 (s, 1H), 10.56 (s, 1H), 8.49 (s, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.65 (d, J=8.4 Hz, 1H), 7.44 (d, J=2.4 Hz, 1H), 4.39-4.34 (m, 1H), 3.92 (s, 3H), 2.51-2.21 (m, 10H), 1.91-1.89 (m, 1H), 1.34-1.32 (m, 1H), 1.06-0.96 (m, 4H), 0.78-0.62 (m, 5H).
N-[3-(4-cyclopropoxy-6-methoxypyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(4-ethylpiperazin-1-yl)methyl]cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2: 14.10 min; LCMS (ESI, m/z): [M+H]+=492.3. 1H NMR (400 MHz, DMSO-d6): δ 11.62 (s, 1H), 10.56 (s, 1H), 8.49 (s, 1H), 7.86 (d, J=8.8 Hz, 1H), 7.65 (d, J=8.8 Hz, 1H), 7.44 (d, J=2.4 Hz, 1H), 4.39-4.34 (m, 1H), 3.92 (s, 3H), 2.51-2.21 (m, 7H), 1.91-1.89 (m, 1H), 1.34-1.32 (m, 1H), 1.06-0.96 (m, 4H), 0.78-0.62 (m, 5H).
To a solution of 4-chloro-3-nitropyridin-2-ol (12.0 g, 68.97 mmol) in toluene (150.0 mL) was added CD3I (20.0 g, 137.94 mmol) and Ag2CO3 (28.6 g, 103.46 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (66/34, v/v) to afford 4-chloro-2-(methoxy-d3)-3-nitropyridine (8.0 g, 60%) as a white solid. LCMS (ESI, m/z): [M+H]+=192.0.
To a solution of 4-chloro-2-(methoxy-d3)-3-nitropyridine (8.0 g, 41.76 mmol) in DMF (100.0 mL) was added cyclopropanol (3.6 g, 62.63 mmol) and Cs2CO3 (20.4 g, 62.63 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (74/26, v/v) to afford 4-cyclopropoxy-2-(methoxy-d3)-3-nitropyridine (4.7 g, 52%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=214.1.
To a solution of 4-cyclopropoxy-2-(methoxy-d3)-3-nitropyridine (4.7 g, 22.04 mmol) in MeOH (30.0 mL)/H2O (6.0 mL) was added NH4Cl (4.7 g, 88.18 mmol) at room temperature. Then Fe (3.7 g, 66.13 mmol) was added to the mixture at 80° C. The resulting mixture was stirred at 80° C. for 3 h. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (78/22, v/v) to afford 4-cyclopropoxy-2-(methoxy-d3)pyridin-3-amine (3.1 g, 76%) as a white solid. LCMS (ESI, m/z): [M+H]+=184.1.
To a solution of 4-cyclopropoxy-2-(methoxy-d3)pyridin-3-amine (3.1 g, 16.92 mmol) in MeCN (35.0 mL) were added CH2I2 (4.5 g, 16.92 mmol) and tert-butyl nitrite (5.2 g, 50.76 mmol) at room temperature. The resulting mixture was stirred at 80 C for 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (82/18, v/v) to afford 4-cyclopropoxy-3-iodo-2-(methoxy-d3)pyridine (2.8 g, 56%) as a white solid. LCMS (ESI, m/z): [M+H]+=295.0.
To a solution of 4-cyclopropoxy-3-iodo-2-(methoxy-d3)pyridine (2.8 g, 9.52 mmol) in 1,4-dioxane (24.0 mL) and H2O (6.0 mL) were added (6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)boronic acid (3.1 g, 9.52 mmol), K2CO3 (3.9 g, 28.56 mmol) and Pd(dppf)Cl2 (1.3 g, 1.90 mmol) at room temperature under N2. The mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (81/19, v/v) to afford 6-chloro-3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (2.0 g, 46%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=449.2.
To a solution of 6-chloro-3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (2.0 g, 4.45 mmol) in THF (20.0 mL) were added Pd2(dba)3 (815.7 mg, 0.89 mmol), XPhos (849.3 mg, 1.78 mmol) and LiHMDS (8.9 mL, 1.0 mol/L) at room temperature under N2. The resulting mixture was stirred at 60° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (87/13, v/v) to afford 3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (1.2 g, 62%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=430.2.
To a solution of 3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (1.2 g, 2.79 mmol) in DMF (15.0 mL) was added trans-2-(methoxycarbonyl)cyclopropane-1-carboxylic acid (483.1 mg, 3.35 mmol), DIEA (1.8 g, 13.97 mmol) and HATU (1.6 g, 4.19 mmol) at 0 C under N2. The resulting mixture was stirred at 0° C. for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (80/20, v/v) to afford methyl (trans)-2-((3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)carbamoyl)cyclopropane-1-carboxylate (1.2 g, 77%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=556.3.
To a solution of methyl (trans)-2-((3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)carbamoyl)cyclopropane-1-carboxylate (1.2 g, 2.16 mmol) in THF (12.0 mL)/CH3OH (3.0 mL) was added NaBH4 (1.6 g, 43.18 mmol) at room temperature. The resulting mixture was stirred at room temperature for 6 h. After the reaction was completed, the reaction mixture was quenched with water at 0° C. and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (55/45, v/v) to afford trans-N-(3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(hydroxymethyl)cyclopropane-1-carboxamide (750.0 mg, 65%) as a white solid. LCMS (ESI, m/z): [M+H]+=528.3.
To a solution of trans-N-(3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(hydroxymethyl)cyclopropane-1-carboxamide (750.0 mg, 1.42 mmol) in CH2Cl2 (10.0 mL) was added Dess-Martin (904.2 mg, 2.13 mmol) at 0 C under N2. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with CH2Cl2. The combined organic layer was washed with NaHCO3 (aq), brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-(3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-formylcyclopropane-1-carboxamide (700.0 mg, crude) as a brown solid. LCMS (ESI, m/z): [M+H]+=526.2.
To a solution of trans-N-(3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-formylcyclopropane-1-carboxamide (700.0 mg, crude) in CH2Cl2 (10.0 mL) were added dimethylamine hydrochloride (325.7 mg, 4.00 mmol) and NaBH3CN (251.0 mg, 4.00 mmol) at 0 C under N2. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum CH2Cl2/CH3OH (10/01, v/v) to afford trans-N-(3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (200.0 mg, 23%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=555.3.
To a solution of trans-N-(3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (200.0 mg, 0.36 mmol) in CH2Cl2 (1.5 mL) was added TFA (1.5 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (1.5 mL) was added NH3·H2O (1.5 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH3CN/H2O (55/45, v/v) to afford trans-N-(3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (50.0 mg, 32%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=425.2.
The product trans-N-(3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide (50.0 mg, 0.12 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRAL ART Cellulose-SC, 2×25 cm, 5 m; Mobile Phase A: Hex: DCM=3:1 (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 10% B to 10% B in 12.5 min; Wave Length: 220/254 nm) to afford N-(3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide Enantiomer 1 (14.4 mg, 57%) as a white solid and N-(3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide Enantiomer 2 (13.3 mg, 52%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 238 and 239 in Table 1.
N-(3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1: 8.39 min; LCMS (ESI, m/z): [M+H]+=425.2. 1H NMR (400 MHz, DMSO-d6): δ 11.50 (d, J=2.0 Hz, 1H), 10.52 (s, 1H), 8.07 (d, J=5.6 Hz, 1H), 7.83 (d, J=8.8 Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 7.34 (d, J=2.4 Hz, 1H), 7.14 (d, J=6.0 Hz, 1H), 3.94-3.91 (m, 1H), 2.35-2.30 (m, 1H), 2.24-2.19 (m, 7H), 1.91-1.88 (m, 1H), 1.37-1.33 (m, 1H), 1.07-1.03 (m, 1H), 0.81-0.76 (m, 2H), 0.74-0.69 (m, 1H), 0.66-0.62 (m, 2H).
N-(3-(4-cyclopropoxy-2-(methoxy-d3)pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((dimethylamino)methyl)cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2: 11.34 min; LCMS (ESI, m/z): [M+H]+=425.2. 1H NMR (400 MHz, DMSO-d6): δ 11.50 (d, J=2.0 Hz, 1H), 10.52 (s, 1H), 8.07 (d, J=5.6 Hz, 1H), 7.83 (d, J=8.8 Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 7.34 (d, J=2.4 Hz, 1H), 7.14 (d, J=6.0 Hz, 1H), 3.94-3.91 (m, 1H), 2.34-2.29 (m, 1H), 2.21-2.18 (m, 7H), 1.91-1.89 (m, 1H), 1.37-1.32 (m, 1H), 1.07-1.03 (m, 1H), 0.81-0.76 (m, 2H), 0.73-0.69 (m, 1H), 0.66-0.62 (m, 2H).
To a solution of trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-formylcyclopropane-1-carboxamide (450.0 mg, 0.87 mmol) in CH2Cl2 (5.0 mL) was added methanamine (0.2 mL, 2 mol/L in THF) and NaBH3CN (162.3 mg, 2.58 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reversed phase flash column chromatography with water/MeOH (60/40, v/v) to afford trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((methylamino)methyl)cyclopropane-1-carboxamide (100.0 mg, 22%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=538.3.
To a solution of trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((methylamino)methyl)cyclopropane-1-carboxamide (130.0 mg, 0.24 mmol) in CH2Cl2 (2.0 mL) was added TFA (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (4.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reversed phase flash column chromatography with water/MeOH (0/100, v/v) to afford trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((methylamino)methyl)cyclopropane-1-carboxamide (70.0 mg, 71%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=408.2.
The compound trans−-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((methylamino)methyl)cyclopropanecarboxamide (70.0 mg, 0.17 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRAL ART Amylose-SA, 2×25 cm, 5 um; Mobile Phase A: Hex: DCM=3: 1 (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 20% B to 20% B in 19 min; 254 nm) to afford N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((methylamino)methyl)cyclopropane-1-carboxamide Enantiomer 1 (5.5 mg, 15%) as a white solid and N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((methylamino)methyl)cyclopropane-1-carboxamide Enantiomer 2 (6.1 mg, 18%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 240 and 241 in Table 1.
N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((methylamino)methyl)cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1: 22.47 min; LCMS (ESI, m/z): [M+H]+=408.1. 1H NMR (400 MHz, DMSO-d6): δ 11.55 (s, 1H), 10.49 (d, J=4.8 Hz, 1H), 8.08 (d, J=5.6 Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 7.35 (s, 1H), 7.14 (d, J=6.0 Hz, 1H), 4.00-3.92 (m, 1H), 3.82 (s, 3H), 2.68-2.60 (m, 1H), 2.34 (s, 3H), 1.96-1.93 (m, 1H), 1.49-1.42 (m, 1H), 1.05-1.03 (m, 1H), 0.88-0.78 (m, 3H), 0.72-0.64 (m, 2H).
N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-((methylamino)methyl)cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2:26.92 min; LCMS (ESI, m/z): [M+H]+=408.1. 1H NMR (400 MHz, DMSO-d6): δ 11.55 (s, 1H), 10.47 (s, 1H), 8.07 (d, J=5.6 Hz, 1H), 7.83 (d, J=8.0 Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 7.35 (s, 1H), 7.14 (d, J=5.6 Hz, 1H), 3.95-3.91 (m, 1H), 3.81 (s, 3H), 2.68-2.60 (m, 1H), 2.32 (s, 3H), 1.97-1.91 (m, 1H), 1.49-1.42 (m, 1H), 1.05-1.02 (m, 1H), 0.81-0.74 (m, 3H), 0.70-0.64 (m, 2H).
To a solution of 4-chloro-3-nitropyridin-2-ol (30.0 g, 171.88 mmol) in toluene (900.0 mL) was added Ag2CO3 (71.1 g, 257.82 mmol) and CH3I (48.8 g, 343.76 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 1 h. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/9, v/v) to afford 4-chloro-2-methoxy-3-nitropyridine (18.7 g, 58%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=189.0.
To a solution of 4-chloro-2-methoxy-3-nitropyridine (18.7 g, 99.17 mmol) in DMF (600.0 mL) was added cyclobutanol (10.7 g, 148.75 mmol) and Cs2CO3 (48.5 g, 148.75 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 1 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/9, v/v) to afford 4-cyclobutoxy-2-methoxy-3-nitropyridine (13.8 g, 62%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=225.1
To a solution of 4-cyclobutoxy-2-methoxy-3-nitropyridine (13.8 g, 61.55 mmol) in methanol/H2O (100.0 mL/100 mL) was added NH4Cl (13.2 g, 246.19 mmol) and Fe (10.3 g, 184.64 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 1 h. After the reaction was completed, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/10, v/v) to afford 4-cyclobutoxy-2-methoxypyridin-3-amine (8.5 g, 71%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=195.1.
To a solution of 4-cyclobutoxy-2-methoxypyridin-3-amine (8.5 g, 43.76 mmol) in CAN (300.0 mL) was added CH2I2 (11.7 g, 43.76 mmol) and t-BuNO2 (20.3 g, 196.93 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 1 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/9, v/v) to afford 4-cyclobutoxy-3-iodo-2-methoxypyridine (4.2 g, 32%) as a brown oil. LCMS (ESI, m/z): [M+H]+=306.0.
To a solution of 4-cyclobutoxy-3-iodo-2-methoxypyridine (2.1 g, 10.94 mmol) in 1,4-dioxane/H2O (100.0 mL/20.0 mL) was added 6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-ylboronic acid (1.9 g, 5.77 mmol), K3PO4 (3.8 g, 17.30 mmol) and PdAMPhos (0.8 g, 1.15 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 1 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/9, v/v) to afford 3-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-4-cyclobutoxy-2-methoxypyridine (2.0 g, 73%) as a brown oil. LCMS (ESI, m/z): [M+H]+=460.2.
To a solution of 3-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-3-yl)-4-cyclobutoxy-2-methoxypyridine (2.0 g, 4.35 mmol) in THE (60.0 mL) was added Pd2(dba)3 (0.4 g, 0.44 mmol), XPhos (0.6 g, 1.30 mmol) and LiHMDS (13.0 mL, 1 mol/L) at room temperature under N2. The resulting mixture was stirred at 80° C. for 1 h. After the reaction was completed, the reaction was quenched by the addition of aq. NH4Cl at 0° C. and then extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (1/4, v/v) to afford 3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (1.7 g, 89%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=441.2.
To a solution of 3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (1.7 g, 3.86 mmol) in DMF (40.0 mL) was added DIEA (2.5 g, 19.29 mmol), trans-2-(methoxycarbonyl)cyclopropane-1-carboxylic acid (0.7 g, 4.63 mmol) and HATU (1.8 g, 4.63 mmol) at 0° C. The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (7/1, v/v) to afford methyl trans-2-{[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}cyclopropane-1-carboxylate (853.0 mg, 46%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=567.3.
To a solution of methyl trans-2-{[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}cyclopropane-1-carboxylate (800.0 mg, 1.41 mmol) in CH3OH/THF (6.0 mL/9.0 mL) was added NaBH4 (534.1 mg, 14.12 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (6/1, v/v) to afford trans-N-[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (651.4 mg, 86%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=539.3.
To a solution of trans-N-[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-(hydroxymethyl)cyclopropane-1-carboxamide (600.0 mg, 1.11 mmol) in CH2Cl2 (18.0 mL) was added Dess-Martin (708.6 mg, 1.67 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. After the reaction was completed, The reaction was diluted with aq·NaHCO3 and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (800.0 mg, crude) as a white oil. LCMS (ESI, m/z): [M+H]+=537.2.
To a solution of trans-N-[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-formylcyclopropane-1-carboxamide (480.0 mg, 0.89 mmol) in CH2Cl2 (15.0 mL) was added dimethylamine hydrochloride (218.8 mg, 2.68 mmol) and NaBH3CN (168.6 mg, 2.68 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (6/1, v/v) to afford trans-N-[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide (400.0 mg, 79%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=566.3.
To a solution of trans-N-[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide (400.0 mg, 0.71 mmol) in DCM (6.0 mL) was added TFA (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The mixture was evaporated under reduced pressure. To the above mixture was added ACN/NH3·H2O (6.0 mL/6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for additional 1 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with CH3CN/H2O (3/7, v/v) to afford trans-N-[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide (75.0 mg, 24%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=436.2.
The product trans-N-[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide (75.0 mg, 0.17 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRAL ART Cellulose-SC, 2×25 cm, 5 m; Mobile Phase A:Hex:DCM=3:1 (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 20 mL/min; Gradient: 15% B to 15% B in 8 min; Wave Length: 220/254 nm) to afford N-[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 1 (7.2 mg, 19%) as a white solid and N-[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 2 (5.8 mg, 15%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 242 and 243 in Table 1.
N-[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1 (min): 5.14; LCMS (ESI, m/z): [M+H]+=436.2. 1H NMR (400 MHz, DMSO-d6): δ 11.52 (s, 1H), 10.53 (s, 1H), 8.00 (d, J=5.6 Hz, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.66 (d, J=8.4 Hz, 1H), 7.41 (d, J=2.4 Hz, 1H), 6.68 (d, J=5.6 Hz, 1H), 4.80-4.76 (m, 1H), 3.81 (s, 3H), 2.43-2.37 (m, 2H), 2.33-2.28 (m, 1H), 2.20-2.15 (m, 6H), 1.98-1.91 (m, 4H), 1.77-1.74 (m, 1H), 1.65-1.60 (m, 1H), 1.37-1.28 (m, 1H), 1.09-1.01 (m, 1H), 0.72-0.68 (m, 1H).
N-[3-(4-cyclobutoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-2-[(dimethylamino)methyl]cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2 (min): 6.57; LCMS (ESI, m/z): [M+H]+=436.2. 1H NMR (400 MHz, DMSO-d6): δ 11.53 (s, 1H), 10.55 (s, 1H), 7.99 (d, J=6.0 Hz, 1H), 7.86 (d, J=8.8 Hz, 1H), 7.66 (d, J=8.8 Hz, 1H), 7.41 (d, J=2.4 Hz, 1H), 6.68 (d, J=5.6 Hz, 1H), 4.79-4.76 (m, 1H), 3.81 (s, 3H), 2.43-2.37 (m, 2H), 2.32-2.27 (m, 1H), 2.20-2.15 (m, 6H), 1.98-1.89 (m, 4H), 1.76-1.74 (m, 1H), 1.65-1.60 (m, 1H), 1.36-1.24 (m, 1H), 1.07-1.02 (m, 1H), 0.73-0.68 (m, 1H).
To a mixture of ethyl trans-2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)cyclopropane-1-carboxylate (0.9 g, 3.52 mmol) and 3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (1.0 g, 2.34 mmol) in THF (40.0 mL) was added AlMe3 (3.5 mL, 2 mol/L) at 0° C. under N2. The resulting mixture was stirred at 80° C. for 2 h. After the reaction was completed, the resulting mixture was quenched with H2O. The mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (10/1, v/v) to afford trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)cyclopropane-1-carboxamide (1.6 g, 87%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=623.3.
To a solution of trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)cyclopropane-1-carboxamide (1.6 g, 2.57 mmol) in MeOH (30.0 mL) was added TsOH (0.4 g, 2.57 mmol) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (5/1, v/v) to afford trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-hydroxyethyl)cyclopropane-1-carboxamide (450.0 mg, 32%) as a brown solid. LCMS (ESI, m/z): [M+H]+=539.3.
To a solution of trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-hydroxyethyl)cyclopropane-1-carboxamide (400.0 mg, 0.74 mmol) in CH2Cl2 (30.0 mL) was added Dess-Martin (472.4 mg, 1.12 mmol) at 0° C. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-oxoethyl)cyclopropane-1-carboxamide (400.0 mg, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=537.3.
To a solution of trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-oxoethyl)cyclopropane-1-carboxamide (400.0 mg, crude) in CH2Cl2 (20.0 mL) was added dimethylamine hydrochloride (182.3 mg, 2.24 mmol) and NaBH3CN (140.5 mg, 2.24 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was quenched with CH3OH. The resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (5/1, v/v) to afford trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide (90.0 mg, 21%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=566.3.
To a solution of trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide (90.0 mg, 0.16 mmol) in CH2Cl2 (2.0 mL) was added TFA (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (2.0 mL) was added NH3·H2O (2.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (5/1, v/v) to afford trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide (50.0 mg, 72%) as a white solid. LCMS (ESI, m/z): [M+H]+=436.2.
The product of trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide (50.0 mg, 0.11 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column CHIRALPAK IE, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 12 mL/min; Gradient: 50% to 50% in 27 min; Wave Length: 220/254 nm) to afford N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide Enantiomer 1 (5.3 mg, 10%) as a white solid and N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide Enantiomer 2 (5.1 mg, 10%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 244 and 245 in Table 1.
N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1 (min): 12.79; LCMS (ESI, m/z): [M+H]+=436.3. 1H NMR (400 MHz, DMSO-d6): δ 11.49 (s, 1H), 10.46 (s, 1H), 8.07 (d, J=6.0 Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 7.55 (d, J=8.8 Hz, 1H), 7.34 (s, 1H), 7.13 (d, J=5.6 Hz, 1H), 4.01-3.93 (m, 1H), 3.81 (s, 3H), 2.33-2.30 (m, 2H), 2.13 (s, 6H), 1.88-1.82 (m, 1H), 1.47-1.37 (m, 2H), 1.24-1.16 (m, 1H), 1.09-0.91 (m, 1H), 0.89-0.75 (m, 2H), 0.73-0.58 (m, 3H).
N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethyl)cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2 (min): 18.32; LCMS (ESI, m/z): [M+H]+=436.3. 1H NMR (400 MHz, DMSO-d6): δ 11.49 (s, 1H), 10.46 (s, 1H), 8.07 (d, J=5.6 Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 7.55 (d, J=8.8 Hz, 1H), 7.34 (s, 1H), 7.14 (d, J=5.6 Hz, 1H), 3.99-3.93 (m, 1H), 3.81 (s, 3H), 2.34-2.31 (m, 2H), 2.13 (s, 6H), 1.89-1.84 (m, 1H), 1.47-1.35 (m, 2H), 1.24-1.16 (m, 1H), 1.12-1.00 (m, 1H), 0.86-0.73 (m, 2H), 0.68-0.64 (m, 3H).
To a solution of 3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-amine (1.8 g, 4.41 mmol) in DMF (30.0 mL) were added 5-(tert-butoxycarbonyl)-5-azaspiro[2.3]hexane-1-carboxylic acid (1.0 g, 4.41 mmol), DIEA (2.3 g, 17.64 mmol) and HATU (2.4 g, 6.17 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the resulting mixture was diluted with water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (3/1, v/v) to afford tert-butyl 1-{[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}-5-azaspiro[2.3]hexane-5-carboxylate (2.2 g, 83%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=609.3.
To a solution of tert-butyl 1-{[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]carbamoyl}-5-azaspiro[2.3]hexane-5-carboxylate (1.0 g, 1.64 mmol) in DCM (10.0 mL) was added HCOOH (20.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with water. The pH value of the mixture was adjusted to 8 with aq·NaHCO3. The resulting mixture was extracted with DCM. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/MeOH (4/1, v/v) to afford N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-5-azaspiro[2.3]hexane-1-carboxamide (0.4 g, 45%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=509.3.
To a solution of N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-5-azaspiro[2.3]hexane-1-carboxamide (550.0 mg, 1.08 mmol) in CH3OH (20.0 mL) was added a solution of HCHO in H2O (1.1 mL, 37%) at room temperature. The resulting mixture was stirred at room temperature for 10 min. Then NaBH3CN (203.8 mg, 3.24 mmol) was added to the mixture. The resulting mixture was stirred at room temperature for 30 min. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with acetonitrile/water (2/3, v/v) to afford N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-5-methyl-5-azaspiro[2.3]hexane-1-carboxamide (130.0 mg, 23%) as a white solid. LCMS (ESI, m/z): [M+H]+=523.3.
To a solution of N-[3-(2,6-dimethoxyphenyl)-1-{[2-(trimethylsilyl)ethoxy]methyl}pyrrolo[2,3-b]pyridin-6-yl]-5-methyl-5-azaspiro[2.3]hexane-1-carboxamide (176.0 mg, 0.34 mmol) in DCM (10.0 mL) was added TFA (10.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 1.5 h. The resulting mixture was concentrated under reduced pressure. To the above mixture was added CH3CN (6.0 mL) and NH3·H2O (6.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography with CH3CN/H2O (2/3, v/v) to afford N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-5-methyl-5-azaspiro[2.3]hexane-1-carboxamide (130.0 mg, 98%) as a white solid. LCMS (ESI, m/z): [M+H]+=393.2.
The racemic N-(3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-5-methyl-5-azaspiro[2.3]hexane-1-carboxamide (130.0 mg, 0.33 mmol) was separated by Chiral-Prep-HPLC with the following conditions (Column: CHIRALPAK IG, 2×25 cm, 5 m; Mobile Phase A: Hex: DCM=1: 1 (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH—HPLC; Flow rate: 17 mL/min; Gradient: 50% B to 50% B in 14.5 min; Wave Length: 220/254 nm) to afford N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-5-methyl-5-azaspiro[2.3]hexane-1-carboxamide Enantiomer 1 (13.5 mg, 13%) as a yellow solid and N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-5-methyl-5-azaspiro[2.3]hexane-1-carboxamide Enantiomer 2 (14.0 mg, 12%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 246 and 247 in Table 1.
N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-5-methyl-5-azaspiro[2.3]hexane-1-carboxamide Enantiomer 1: Retention Time 1: 4.65 min; LCMS (ESI, m/z): [M+H]+=393.2. 1H NMR (400 MHz, DMSO-d6): δ 11.41 (s, 1H), 10.50 (s, 1H), 7.80 (d, J=8.4 Hz, 1H), 7.52 (d, J=8.4 Hz, 1H), 7.30-7.27 (m, 2H), 6.76-6.72 (m, 2H), 3.67 (s, 6H), 3.36-3.24 (m, 4H), 2.30 (s, 3H), 2.13-2.10 (m, 1H), 1.10-1.04 (m, 2H).
N-[3-(2,6-dimethoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-6-yl]-5-methyl-5-azaspiro[2.3]hexane-1-carboxamide Enantiomer 2: Retention Time 2: 10.92 min; LCMS (ESI, m/z): [M+H]+=393.2. 1H NMR (400 MHz, DMSO-d6): δ 11.41 (s, 1H), 10.49 (s, 1H), 7.80 (d, J=8.4 Hz, 1H), 7.53 (d, J=8.4 Hz, 1H), 7.30-7.22 (m, 2H), 6.76-6.70 (m, 2H), 3.72 (s, 6H), 3.34-3.21 (m, 4H), 2.29 (s, 3H), 2.15-2.02 (m, 1H), 1.12-1.03 (m, 2H).
To a solution of trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-formylcyclopropane-1-carboxamide (670.0 mg, 1.28 mmol) in CH2Cl2 (20.0 mL) was added dimethyl-D6-amine hydrochloride (336.8 mg, 3.85 mmol) and NaBH3CN (241.7 mg, 3.87 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction was quenched with H2O and extracted with dichloromethane. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with H2O/CH3CN (3/7, v/v) to afford trans-2-((bis(methyl-d3)amino)methyl)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (200.0 mg, 27%) as a green solid. LCMS (ESI, m/z): [M+H]+=558.3.
To a solution of trans-2-((bis(methyl-d3)amino)methyl)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (300.0 mg, 0.54 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for another 2 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with H2O/CH3CN (1/1, v/v) to afford trans-2-((bis(methyl-d3)amino)methyl)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (190.0 mg, 82%) as a white solid. LCMS (ESI, m/z): [M+H]+=428.3.
The product trans-2-((bis(methyl-d3)amino)methyl)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (190.0 mg, 0.34 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRAL ART Cellulose-SC, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: MeOH: EtOH=1: 1—HPLC; Flow rate: 20 mL/min; Gradient: 20% to 20% in 17 min; Wave Length: 220/254 nm) to afford 2-((bis(methyl-d3)amino)methyl)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide Enantiomer 1 (44.7 mg, 60%) as a white solid and 2-((bis(methyl-d3)amino)methyl)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide Enantiomer 2 (30.1 mg, 40%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 248 and 249 in Table 1.
2-((bis(methyl-d3)amino)methyl)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide Enantiomer 1: Retention Time 1 (min): 11.48; LCMS (ESI, m/z): [M+H]+=428.3. 1H NMR (400 MHz, DMSO-d6): δ 11.50 (s, 1H), 10.52 (s, 1H), 8.07 (d, J=5.6 Hz, 1H), 7.83 (d, J=8.8 Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 7.34 (d, J=2.0 Hz, 1H), 7.14 (d, J=5.6 Hz, 1H), 3.94-3.91 (m, 1H), 3.82 (s, 3H), 2.32-2.28 (m, 1H), 2.21-2.16 (m, 1H), 1.91-1.89 (m, 1H), 1.38-1.30 (m, 1H), 1.07-1.03 (m, 1H), 0.87-0.76 (m, 2H), 0.73-0.64 (m, 3H).
2-((bis(methyl-d3)amino)methyl)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide Enantiomer 2: Retention Time 2 (min): 15.63; LCMS (ESI, m/z): [M+H]+=428.4. 1H NMR (400 MHz, DMSO-d6): δ 11.50 (s, 1H), 10.52 (s, 1H), 8.07 (d, J=6.0 Hz, 1H), 7.83 (d, J=8.8 Hz, 1H), 7.55 (d, J=8.4 Hz, 1H), 7.34 (d, J=2.0 Hz, 1H), 7.14 (d, J=5.6 Hz, 1H), 3.99-3.91 (m, 1H), 3.81 (s, 3H), 2.29-2.25 (m, 1H), 2.18-2.13 (m, 1H), 1.90-1.88 (m, 1H), 1.37-1.32 (m, 1H), 1.08-1.02 (m, 1H), 0.81-0.76 (m, 2H), 0.71-0.63 (m, 3H).
To a solution of tert-butyl N-(5-bromo-4-methoxypyridin-2-yl)carbamate (635.0 mg, 2.10 mmol) in THF (20.0 mL) was added NaH (251.3 mg, 60%) at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h under N2. Then methyl iodide (1.5 g, 10.47 mmol) was added dropwise to the mixture at 0° C. under N2. The resulting mixture was stirred at 0° C. for 1 h. After the reaction was completed, the reaction mixture was quenched with water and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford tert-butyl N-(5-bromo-4-methoxypyridin-2-yl)-N-methylcarbamate (500.0 mg, 75%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=317.0.
To a solution of tert-butyl N-(5-bromo-4-methoxypyridin-2-yl)-N-methylcarbamate (460.0 mg, 1.45 mmol) in dioxane/H2O (10.0 mL/2.0 mL) was added 6-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridine (592.9 mg, 1.45 mmol), K2CO3 (601.3 mg, 4.35 mmol) and Pd(dppf)Cl2 (118.4 mg, 0.15 mmol) at room temperature under N2. The resulting mixture was stirred at 80° C. for 16 h. After the reaction was completed, the reaction was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford tert-butyl N-[5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxypyridin-2-yl]-N-methylcarbamate (278.0 mg, 37%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=519.2.
To a solution of tert-butyl N-[5-(6-chloro-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxypyridin-2-yl]-N-methylcarbamate (258.0 mg, 0.50 mmol) in 1,4-dioxane (8.0 mL) was added cyclopropanecarboxamide (211.5 mg, 2.49 mmol), BrettPhos (53.4 mg, 0.10 mmol), Cs2CO3 (485.8 mg, 1.49 mmol) and BrettPhos Pd G3 (45.1 mg, 0.05 mmol) at room temperature under N2. The resulting mixture was stirred with microwave at 120° C. for 1.5 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (2/1, v/v) to afford tert-butyl (5-(6-(cyclopropanecarboxamido)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-4-methoxypyridin-2-yl)(methyl)carbamate (168.0 mg, 72%) as a white oil. LCMS (ESI, m/z): [M+H]+=568.3.
To a solution of tert-butyl N-[5-(6-cyclopropaneamido-1-[[2-(trimethylsilyl)ethoxy]methyl]pyrrolo[2,3-b]pyridin-3-yl)-4-methoxypyridin-2-yl]-N-methylcarbamate (168.0 mg, 0.30 mmol) in DCM (3.0 mL) was added TFA (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (3.0 mL) was added NH3·H2O (3.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC with the following conditions Column: (XBridge Prep OBD C18 Column, 30×150 mm, 5 um; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 23% B to 53% B in 7 min; 254 nm) to afford N-[3-[4-methoxy-6-(methylamino)pyridin-3-yl]-1H-pyrrolo[2,3-b]pyridin-6-yl]cyclopropanecarboxamide (16.1 mg, 16%) as a white solid. LCMS (ESI, m/z): [M+H]+=338.2. 1H NMR (300 MHz, DMSO-d6): δ 11.39 (s, 1H), 10.52 (s, 1H), 8.00 (s, 1H), 7.91-7.82 (m, 2H), 7.36 (d, J=2.4 Hz, 1H), 6.38-6.34 (m, 1H), 6.10 (s, 1H), 3.78 (s, 3H), 2.81 (d, J=4.8 Hz, 3H), 2.07-1.99 (m, 1H), 0.82-0.74 (m, 4H).
To a solution of 2-(vinyloxy)ethan-1-ol (20.0 g, 227.0 mmol) in DCM (200.0 mL) was added TEA (68.9 g, 681.00 mmol), DMAP (2.8 g, 22.70 mmol) and tert-butylchlorodiphenylsilane (74.9 g, 272.40 mmol) at room temperature under N2. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetate (10/1, v/v) to afford tert-butyldiphenyl(2-(vinyloxy)ethoxy)silane (40.0 g, 53%) as a yellow oil.
To a mixture of Rh2(OAc)4 (0.2 g, 0.46 mmol) in tert-butyldiphenyl(2-(vinyloxy)ethoxy)silane (5.0 g, 15.31 mmol) was added dropwise ethyl 2-diazoacetate (8.7 g, 76.57 mmol) at room temperature under N2 in 10 h. The resulting mixture was stirred at room temperature for 16 h under N2. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with petroleum ether/ethyl acetated (10/1, v/v) to afford ethyl (trans)-2-(2-((tert-butyldiphenylsilyl)oxy)ethoxy)cyclopropane-1-carboxylate (2.3 g, 36%) as a colorless oil. LCMS (ESI, m/z): [M+H]+=413.2.
To a solution of ethyl (trans)-2-(2-((tert-butyldiphenylsilyl)oxy)ethoxy)cyclopropane-1-carboxylate (2.2 g, 5.33 mmol) in DMF (20.0 mL) was added TEA (1.6 g, 16.00 mmol) and TBAF (4.2 g, 16.00 mmol) at room temperature. The resulting mixture was stirred at 80° C. for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford ethyl (trans)-2-(2-hydroxyethoxy)cyclopropane-1-carboxylate (1.3 g, crude) as a colorless oil. LCMS (ESI, m/z): [M+H]+=175.1.
To a solution of ethyl (trans)-2-(2-hydroxyethoxy)cyclopropane-1-carboxylate (1.2 g, crude) in CH2Cl2 (30.0 mL) was added Dess-Martin (4.4 g, 10.33 mmol) at 0° C. The resulting mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction mixture was diluted with H2O and extracted with CH2Cl2. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford ethyl (trans)-2-(2-oxoethoxy)cyclopropane-1-carboxylate (1.1 g, crude) as a yellow solid. LCMS (ESI, m/z): [M+H]+=173.1.
To a solution of ethyl (trans)-2-(2-oxoethoxy)cyclopropane-1-carboxylate (1.1 g, crude) in CH2Cl2 (20.0 mL) was added (CH3)2NH in THF (9.6 mL, 2 mol/L) and NaBH3CN (1.2 g, 19.17 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was quenched with CH3OH. The resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (10/1, v/v) to afford ethyl (trans)-2-(2-(dimethylamino)ethoxy)cyclopropane-1-carboxylate (300.0 mg, 23%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=202.1.
To a solution of ethyl (trans)-2-(2-(dimethylamino)ethoxy)cyclopropane-1-carboxylate (471.8 mg, 2.34 mmol) in THE (20.0 mL) was added 3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (500.0 mg, 1.17 mmol) and LiHMDS (3.5 mL, 1 mol/L) at 0° C. under N2. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was quenched with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (10/1, v/v) to afford (trans)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethoxy)cyclopropane-1-carboxamide (180.0 mg, 26%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=582.3.
To a solution of (trans)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethoxy)cyclopropane-1-carboxamide (500.0 mg, 0.86 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with H2O/CH3OH (2/3, v/v) to afford trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethoxy)cyclopropane-1-carboxamide (220.0 mg, 56%) as a white solid. LCMS (ESI, m/z): [M+H]+=452.2.
The product of trans-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethoxy)cyclopropane-1-carboxamide (220.0 mg, 0.49 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IG, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: MeOH: EtOH=1:1—HPLC; Flow rate: 20 mL/min; Gradient: 80% to 80% in 21 min; Wave Length: 220/254 nm; RT1 (min): 10.36; RT2 (min): 15.92) to afford N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethoxy)cyclopropane-1-carboxamide Enantiomer 1 (RT1: 10.36 min, 58.7 mg, 53%) as a white solid and N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethoxy)cyclopropane-1-carboxamide Enantiomer 2 (RT2: 15.92 min, 57.5 mg, 52%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 251 and 252 in Table 1.
N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethoxy)cyclopropane-1-carboxamide Enantiomer 1: RT1 (min): 10.36; LCMS (ESI, m/z): [M+H]+=452.3. 1H NMR (400 MHz, DMSO-d6): δ 11.53 (s, 1H), 10.61 (s, 1H), 8.08 (d, J=5.6 Hz, 1H), 7.82 (d, J=8.4 Hz, 1H), 7.56 (d, J=8.8 Hz, 1H), 7.35 (d, J=2.4 Hz, 1H), 7.14 (d, J=6.0 Hz, 1H), 3.95-3.91 (m, 1H), 3.81 (s, 3H), 3.64-3.52 (m, 3H), 2.61-2.53 (m, 1H), 2.22-2.18 (m, 7H), 1.16-1.13 (m, 2H), 0.82-0.78 (m, 2H), 0.68-0.59 (m, 2H).
N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)-2-(2-(dimethylamino)ethoxy)cyclopropane-1-carboxamide Enantiomer 2: RT2 (min): 15.92; LCMS (ESI, m/z): [M+H]+=452.3. 1H NMR (400 MHz, DMSO-d6): δ 11.53 (s, 1H), 10.61 (s, 1H), 8.08 (d, J=5.6 Hz, 1H), 7.82 (d, J=8.4 Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 7.35 (d, J=2.4 Hz, 1H), 7.14 (d, J=6.0 Hz, 1H), 3.95-3.91 (m, 1H), 3.81 (s, 3H), 3.63-3.52 (m, 3H), 2.49-2.47 (m, 2H), 2.21-2.15 (m, 7H), 1.16-1.13 (m, 2H), 0.82-0.73 (m, 2H), 0.68-0.59 (m, 2H).
To a solution of ethyl (trans)-2-(2-oxoethoxy)cyclopropane-1-carboxylate (1.1 g, 6.39 mmol) in CH2Cl2 (20.0 mL) was added azetidine (1.1 g, 19.17 mmol) and NaBH3CN (1.2 g, 19.17 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. After the reaction was completed, the resulting mixture was quenched with CH3OH. The resulting mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (10/1, v/v) to afford ethyl (trans)-2-(2-(azetidin-1-yl)ethoxy)cyclopropane-1-carboxylate (500.0 mg, 36%) as a yellow oil. LCMS (ESI, m/z): [M+H]+=214.1.
To a solution of ethyl (trans)-2-(2-(azetidin-1-yl)ethoxy)cyclopropane-1-carboxylate (440.0 mg, 2.06 mmol) in THE (20.0 mL) was added 3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-amine (220.0 mg, 0.52 mmol) and LiHMDS (1.6 mL, 1 mol/L) at 0° C. under N2. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with CH2Cl2/CH3OH (10/1, v/v) to afford (trans)-2-(2-(azetidin-1-yl)ethoxy)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (130.0 mg, 42%) as a yellow solid. LCMS (ESI, m/z): [M+H]+=594.3.
To a solution of (trans)-2-(2-(azetidin-1-yl)ethoxy)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (180.0 mg, 0.30 mmol) in CH2Cl2 (5.0 mL) was added TFA (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure. To the residue in CH3CN (5.0 mL) was added NH3·H2O (5.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 4 h. After the reaction was completed, the resulting mixture was diluted with H2O and extracted with ethyl acetate. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash column chromatography with H2O/CH3CN (3/2, v/v) to afford trans-2-(2-(azetidin-1-yl)ethoxy)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (49.0 mg, 34%) as a white solid. LCMS (ESI, m/z): [M+H]+=464.2.
The product of trans-2-(2-(azetidin-1-yl)ethoxy)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide (48.0 mg, 0.10 mmol) was separated by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IG, 2×25 cm, 5 m; Mobile Phase A: Hex (0.5% 2M NH3-MeOH)—HPLC, Mobile Phase B: EtOH: DCM=1: 1—HPLC; Flow rate: 20 mL/min; Gradient: 45% B to 45% B in 24.5 min; Wave Length: 220/254 nm; RT1 (min): 9.43; RT2 (min): 17.99) to afford 2-(2-(azetidin-1-yl)ethoxy)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide Enantiomer 1 (12.8 mg, 53%) as a white solid and 2-(2-(azetidin-1-yl)ethoxy)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide Enantiomer 2 (11.6 mg, 48%) as a white solid. The absolute stereochemistry of Enantiomers 1 and 2 was not assigned. The two enantiomeric structures that could be obtained from chiral separation of the enantiomeric mixture as described above are shown as Compounds 254 and 255 in Table 1.
2-(2-(azetidin-1-yl)ethoxy)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide Enantiomer 1: RT1 (min): 9.43; LCMS (ESI, m/z): [M+H]+=464.2. 1H NMR (400 MHz, DMSO-d6): δ 11.54 (s, 1H), 10.61 (s, 1H), 8.08 (d, J=6.0 Hz, 1H), 7.82 (d, J=8.8 Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 7.35 (d, J=2.4 Hz, 1H), 7.14 (d, J=5.6 Hz, 1H), 3.94-3.92 (m, 1H), 3.82 (s, 3H), 3.58-3.45 (m, 4H), 3.19-3.13 (m, 4H), 2.24-2.18 (m, 1H), 1.99-1.92 (m, 2H), 1.14-1.11 (m, 2H), 0.81-0.77 (m, 2H), 0.67-0.63 (m, 2H).
2-(2-(azetidin-1-yl)ethoxy)-N-(3-(4-cyclopropoxy-2-methoxypyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)cyclopropane-1-carboxamide Enantiomer 2: RT2 (min): 17.99; LCMS (ESI, m/z): [M+H]+=464.3. 1H NMR (400 MHz, DMSO-d6): δ 11.54 (s, 1H), 10.61 (s, 1H), 8.08 (d, J=5.6 Hz, 1H), 7.82 (d, J=8.8 Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 7.35 (d, J=2.4 Hz, 1H), 7.14 (d, J=6.0 Hz, 1H), 3.94-3.92 (m, 1H), 3.82 (s, 3H), 3.52-3.44 (m, 3H), 3.18-3.11 (m, 4H), 2.24-2.18 (m, 1H), 1.98-1.91 (m, 2H), 1.14-1.11 (m, 2H), 0.81-0.77 (m, 2H), 0.74-0.64 (m, 2H).
K562 and HL60 cells, cultured in Iscove's Modified Dulbecco's Media (IMDM) supplemented with 10% FBS, were harvested at 50-80% confluency and plated at 2,000 cells per well (K562) or 1,500 cells per well (HL60) in 384-well tissue culture plates. A subset of wells contained media only (low control, LC). Compounds were serially diluted in DMSO. 40 nL of compound or DMSO only (high control, HC) were added to each well using an Echo 550 liquid handler (Labcyte). The plates were placed in a 37° C. incubator with 5% CO2 for 72 hours. Cell viability was measured using a CellTiter-Glo luminescent cell viability assay (Promega), which allows for relative quantification of metabolically active cells by luminescence-based measurements of intracellular ATP concentrations. Briefly, plates were removed from the incubator and equilibrated for 15 minutes at room temperature prior to the addition of 40 μL of CellTiter-Glo reagent. Plates were then incubated for 30 minutes at room temperature. Luminescence was measured using an EnSpire plate reader (Perkin Elmer). As noted above, luminescence values from wells with DMSO only or media without cells were used as high and low controls (HC and LC), respectively. Normalized percent viability was calculated as follows: percent viability=100×(Lumsample−LumLC)/(LumHC−LumLC). IC50 values were calculated using the XLFit software and are shown in Table 2.
Kinase activity of ABL1 was measured using the ADP-Glo system (Promega), which measures formation of ADP using a luminescence-based method. Compounds were serially diluted in DMSO. 20 nL of compound or DMSO only (high control, HC) were added to a 384-well plate (OptiPlate-384, PerkinElmer) using an Echo550 liquid handler (Labcyte). 15 μL of kinase solution (10 mM MgCl2, 0.01% Brij-35, 2 mM DTT, 0.05% BSA, 1 mM EGTA, 50 mM HEPES pH 7.5, and 3.325 nM ABL1 [Carna Biosciences]) were added to each well of the 384-well plate containing the compounds. No enzyme control wells were included (low control, LC). The plate was incubated at room temperature for 30 minutes. 5 μL of a second solution containing 10 mM MgCl2, 0.01% Brij-35, 2 mM DTT, 0.05% BSA, 1 mM EGTA, 50 mM HEPES pH 7.5, 6 μM Peptide 2 (Perkin Elmer, Cat No 760346), and 1.2 mM ATP were added to each well to start the kinase reaction. The plate was incubated for 90 minutes at room temperature. 20 μL of ADP-Glo reagent (Promega) were then added to each well and the plate was incubated for 40 minutes at room temperature. 40 μL of kinase detection reagent (Promega) was added to each well and the plate was incubated for an additional 45 minutes at room temperature. During this step, ADP was converted to ATP, a substrate for luciferase, to produce luminescence signal. Luminescence was measured on an Envision plate reader (Perkin Elmer). Luminescence signal positively correlates with kinase activity. The percent kinase activity was calculated as follows: percent kinase activity=100×(Lumsample−LumLC)/(LumHC−LumLC). As noted above, DMSO only and no enzyme wells were used as high and low controls, respectively. IC50 values were calculated using the XLFit software.
IC50 data obtained using the screening procedures described above for certain compounds disclosed herein are listed in Table 3.
pCRKL ELISA assay:
K562 or Ba/F3 ABL T315I cells (2.0*105 cells/100 μl/well) were seeded in 96 well (Corning, cat #3799). Compounds were dissolved in DMSO, serially diluted in DMSO and then were added, mixed, and incubated for 90 minutes at 37° C., 5% CO2. Following the 90-minute incubation, plates were centrifuged for 5 min at 3000 RPM and supernatant was removed from each well. Cells were washed 3 times with 150 μl PBS prior to addition of 100 μl cell RIPA lysis buffer (Boston BioProducts, cat #BP-115D) supplied with 1×complete ULTRA cocktail inhibitor (Roche, 05892791001) and 1×PhosSTOP Phosphatase Inhibitor Cocktail Tablets (Roche, 04906837001). Cells were incubated with lysis buffer for 1 hour at 4° C. prior to storage at −80 C.
A capture antibody able to detect phosphorylated and non-phosphorylated CRKL (R&D Systems, cat #AF5127) was added to Meso Scale Discovery (MSD) standard bind plates (MSD, cat #L15XA-3) at 5 ug/mL and incubated at 4° C. overnight. The next day, plates were washed with PBS+0.05% Tween20 (PBST) and 150 μl of 5% BSA blocking solution was added for 1 hour at room temperature with shaking. Plates were washed with PBST. Lysates were thawed and 30 μl of lysate was added to the MSD plates and incubated for 2 hours at room temperature with shaking. MSD plates were washed with PBST and 30 μl of a detection antibody that binds phosphorylated pCRKL (R&D Systems, cat #MAB6910) was added at 1 μg/mL to each well. Plates were incubated for 1 hour at room temperature with shaking. Plates were washed with PBST prior to addition of 30 μl of a sulfo tagged goat anti mouse detection antibody (MSD cat #R32AC-1). Plates were incubated for 1 hour at room temperature with shaking. Plates were washed with PBST prior to addition of 150 ul of 1×MSD read buffer T (MSD, cat 4R92TC-2). Electrochemiluminescence (ECLU) was read on an MSD plate reader (Meso Scale Discovery). The remaining activity by calculated as follows: 00 Relative activity=100×(ELCUsample−ECLULC)/(ECLUHC−ECLULC). The low and high controls (LC/HC) are generated from lysate from wells without cells or with cells treated with 0.1% DMSO, respectively. IC50 values were calculated using XLFit software using a nonlinear regression model with a sigmoidal dose response and are shown in Table 4 and Table 5 below.
Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it is apparent to those skilled in the art that certain minor changes and modifications will be practiced in light of the above teaching. Therefore, the description and examples should not be construed as limiting the scope of the invention.
This application claims priority to and the benefit of U.S. Provisional Application No. 63/087,780, filed on Oct. 5, 2020, the disclosure of which is incorporated herein by reference in its entirety.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US2021/071692 | 10/4/2021 | WO |
| Number | Date | Country | |
|---|---|---|---|
| 63087780 | Oct 2020 | US |
