Patent Application
20250115600
The instant application contains a Sequence Listing which has been submitted electronically in XML file format and is hereby incorporated by reference in its entirety. Said XML copy, created on May 15, 2023, is named 61402-704_601_SL.xml and is 12,141 bytes in size.
The AKT or Protein Kinase B (PKB) family of serine/threonine protein kinases is comprised of 3 highly homologous members, AKT1, AKT2 and AKT3. The family of AKT proteins are involved in signal transduction pathways that regulate cellular processes including apoptosis, proliferation, differentiation and metabolism. The AKT1 pathway is the most frequently deregulated signaling pathways in human cancers. Enhanced activation of all the isoforms can be implicated in tumor development and progression, and has been demonstrated in breast, ovarian, pancreatic, and prostate cancers among others (Song et al., 2019). In cancer cells, AKT1 is involved in proliferation and growth, promoting tumor initiation and suppressing apoptosis, whereas AKT2 regulates cytoskeleton dynamics, favoring local tissue invasion and metastasis. The role of AKT3 hyperactivation in cancer is hypothesized to be involved with possible stimulation of cell proliferation (Hinz, N. et al. Cell Commun Signal 2019, 17(1), 154; Pascual, J. et al. Ann. Oncol. 2019, 30(7), 1051-1060). Expression of these AKT family members is altered in many human malignant carcinomas including gastric, breast, prostate, ovarian and pancreatic. AKT family members are rarely mutated however, the most common mutation is AKT1 E17K which has been reported in 6-8% of breast cancers, 2-6% of colorectal cancers, and in 6% of meningiomas, in human (Yu, Y., et al. PLoS One 2015, 10 (10) No. e0140479). Thus, there is a need to develop new treatments for the modulation of AKT1 and mutants thereof.
In one aspect, the present disclosure provides an AKT1 protein covalently bound to a compound, wherein the compound is covalently bound to a lysine residue of the AKT1 protein. In some embodiments, the compound is an exogenous AKT1 modulator. In some embodiments, the compound is an exogenous AKT1 inhibitor. In some embodiments, the AKT1 protein comprises a E17K mutation, a E40K mutation, or a E49K mutation. In some embodiments, the lysine residue is selected from K17, K40, K49, K158, K163, K179, K267, and K297. In some embodiments, the lysine residue is K17. In some embodiments, the lysine residue is selected from K40, K49, K158, K163, K179, K267, and K297. In some embodiments, the AKT1 protein is in vivo. In some embodiments, the AKT1 protein is an in vivo engineered AKT1 protein, wherein the in vivo engineered AKT1 protein is generated by contacting the AKT1 protein in vivo with the compound. In some embodiments, the in vivo engineered AKT1 protein is a human in vivo engineered AKT1 protein. In some embodiments, the covalent bond between the compound and the lysine residue is a reversible covalent bond. In some embodiments, the reversible covalent bond in the in vivo AKT1 protein is a carbon-nitrogen double bond. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K17 and an aldehyde functional group on the compound. In some embodiments, the aldehyde functional group is an aromatic aldehyde.
In another aspect, the present disclosure provides an in vivo engineered AKT1 protein comprising a non-naturally occurring reversible covalent modification at a lysine residue, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and a lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue. In some embodiments, the AKT1 protein comprises a E17K mutation. In some embodiments, the lysine residue is selected from K17, K40, K49, K158, K163, K179, K267, and K297. In some embodiments, the lysine residue is K17. In some embodiments, the lysine is selected from K40, K49, K158, K163, K179, K267, and K297. In some embodiments, the in vivo engineered AKT1 protein is a human in vivo engineered AKT1 protein.
In another aspect, the present disclosure provides a method of covalently modifying an AKT1 protein, comprising contacting the AKT1 protein with an exogenous AKT1 modulator, wherein the AKT1 modulator comprises a reversible electrophilic moiety thereby forming a reversible covalent AKT1 adduct. In some embodiments, the contacting is in vitro. In some embodiments, the contacting is in vivo. In some embodiments, the AKT1 modulator is an AKT1 inhibitor. In some embodiments, the reversible electrophilic moiety is an aromatic aldehyde.
In another aspect, the present disclosure provides a method of attenuating AKT1 activity, comprising contacting AKT1 protein with an AKT1 inhibitor, wherein the AKT1 inhibitor comprises a reversible electrophilic moiety. In some embodiments, the contacting is in vitro. In some embodiments, following the contacting, the AKT1 activity is attenuated by 50% or more relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by 70% or more relative to a control in the absence of the exogenous AKT1 inhibitor.
In another aspect, the present disclosure provides a compound of Formula (I):
In some embodiments, the compound or salt of Formula (I) is a compound represented by the structure of Formula (I-A):
In another aspect, the present disclosure provides a compound of Formula (II):
In some embodiments, the compound or salt of Formula (II) is a compound represented by the structure of Formula (II-A):
or a pharmaceutically acceptable salt thereof.
In some embodiments, the compound or salt of Formula (II) is a compound represented by the structure of Formula (II-B):
or a pharmaceutically acceptable salt thereof.
In some embodiments, the compound or salt of Formula (II) is a compound represented by the structure of Formula (II-C):
or pharmaceutically acceptable salt thereof.
In some embodiments, the compound is a compound of Table 1, or a salt of any one thereof.
In another aspect, the present disclosure provides a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof.
In another aspect, the present disclosure provides a method of modulating activity of mutant AKT1 comprising, administering to a subject in need thereof a compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of a compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof.
In another aspect, the present disclosure provides a method of selectively modulating activity a mutant AKT1 over a wild type AKT comprising administering to a subject in need thereof a compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of a compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof, wherein the wild type AKT is selected from wild type AKT1 and wild type AKT2.
In another aspect, the present disclosure provides a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of a compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof. In some embodiments, the cancer is selected from breast cancer, colorectal cancer, and meningioma. In some embodiments, the administration modulates activity of a mutant AKT1. In some embodiments, the mutant AKT1 is AKT1 E17K.
All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.
The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:
While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this invention belongs. All patents and publications referred to herein are incorporated by reference.
As used in the specification and claims, the singular form “a”, “an”, and “the” includes plural references unless the context clearly dictates otherwise.
In this application, the use of “or” means “and/or” unless stated otherwise. The terms “and/or” and “any combination thereof” and their grammatical equivalents as used herein, can be used interchangeably. These terms can convey that any combination is specifically contemplated. Solely for illustrative purposes, the following phrases “A, B, and/or C” or “A, B, C, or any combination thereof” can mean “A individually; B individually; C individually; A and B; B and C; A and C; and A, B, and C.” The term “or” can be used conjunctively or disjunctively, unless the context specifically refers to a disjunctive use.
The term “about” or “approximately” can mean within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, “about” can mean within 1 or more than 1 standard deviation, per the practice in the art. Alternatively, “about” can mean a range of up to 20%, up to 15%, up to 10%, up to 5%, or up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, within 5-fold, or within 2-fold, of a value.
Ranges provided herein are understood to be shorthand for all of the values within the range. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50, as well as all intervening decimal values between the aforementioned integers such as, for example, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, and 1.9. With respect to sub-ranges, “nested sub-ranges” that extend from either end point of the range are specifically contemplated. For example, a nested sub-range of an exemplary range of 1 to 50 may comprise 1 to 10, 1 to 20, 1 to 30, and 1 to 40 in one direction, or 50 to 40, 50 to 30, 50 to 20, and 50 to 10 in the other direction.
As used in this specification and claim(s), the words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”) or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. It is contemplated that any embodiment discussed in this specification can be implemented with respect to any method or composition of the present disclosure, and vice versa. Furthermore, compositions of the present disclosure can be used to achieve methods of the present disclosure.
Reference in the specification to “some embodiments,” “an embodiment,” “one embodiment” or “other embodiments” means that a particular feature, structure, or characteristic described in connection with the embodiments is included in at least some embodiments, but not necessarily all embodiments, of the present disclosures. To facilitate an understanding of the present disclosure, a number of terms and phrases are defined below. As used herein, the terms “treatment,” “treat,” and “treating” are meant to include the full spectrum of intervention for the cancer from which the subject is suffering, such as administration of the combination to alleviate, slow, stop, or reverse one or more symptoms of the cancer and to delay the progression of the cancer even if the cancer is not actually eliminated. Treatment can include, for example, a decrease in the severity of a symptom, the number of symptoms, or frequency of relapse, e.g., the inhibition of tumor growth, the arrest of tumor growth, or the regression of already existing tumors.
The term “exogenous” refers to any material introduced from or originating from outside a cell, a tissue or an organism that is not produced by or does not originate from the same cell, tissue, or organism in which it is being introduced. By way of example a chemical compound, nucleic acid or antibody.
The term “nucleic acid” refers to a deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), or a combination thereof, in either a single- or double-stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogues of natural nucleotides that have similar binding properties as the reference nucleotides. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses complementary sequences as well as the sequence explicitly indicated. In some embodiments of any of the nucleic acids described herein, the nucleic acid is DNA. In some embodiments of any of the nucleic acids described herein, the nucleic acid is RNA.
The term “N-terminally positioned” when referring to a position of a first domain or sequence relative to a second domain or sequence in a polypeptide primary amino acid sequence means that the first domain is located closer to the N-terminus of the polypeptide primary amino acid sequence. In some embodiments, there may be additional sequences and/or domains between the first domain or sequence and the second domain or sequence.
The term “C-terminally positioned” when referring to a position of a first domain or sequence relative to a second domain or sequence in a polypeptide primary amino acid sequence means that the first domain is located closer to the C-terminus of the polypeptide primary amino acid sequence. In some embodiments, there may be additional sequences and/or domains between the first domain or sequence and the second domain or sequence.
The term “antibody” is used herein in the broadest sense and encompasses monoclonal antibodies, polyclonal antibodies, dimers, multimers, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit the desired biological activity (Miller et al., J Immunol. 2003, 170, 4854-4861). Antibodies may be murine, human, humanized, chimeric, or derived from other species. (Janeway, C., Travers, P., Walport, M., Shlomchik (2001) Immunol. Biology, 5th Ed., Garland Publishing, New York). Antibodies bound to various types of molecules, such as polyethylene glycols (PEGs), may be used as modified antibodies. Methods for modifying antibodies are already established in the art.
The AKT1 protein described herein includes the naturally occurring (e.g., homo sapien) protein and variants thereof (e.g., Wild-type (WT) AKT1 protein, and mutant AKT1 variants, such as but not limited to E17K AKT1 mutant protein). In some embodiments, the AKT1 protein as described herein can be found in the NCBI database as GenBank: AAL55732.1 (https://www.ncbi.nlm.nih.gov/protein/AAL55732.1).
Modifications can be introduced into a nucleotide sequence by standard techniques known in the art, such as site-directed mutagenesis and polymerase chain reaction (PCR)-mediated mutagenesis. Conservative amino acid substitutions are ones in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., arginine, lysine and histidine), acidic side chains (e.g., aspartic acid and glutamic acid), uncharged polar side chains (e.g., asparagine, cysteine, glutamine, glycine, serine, threonine, tyrosine, and tryptophan), nonpolar side chains (e.g., alanine, isoleucine, leucine, methionine, phenylalanine, proline, and valine), beta-branched side chains (e.g., isoleucine, threonine, and valine), and aromatic side chains (e.g., histidine, phenylalanine, tryptophan, and tyrosine), and aromatic side chains (e.g., histidine, phenylalanine, tryptophan, and tyrosine).
The terms “polypeptide,” “peptide” and “protein” are often used interchangeably herein in reference to a polymer of amino acid residues. A protein, generally, refers to a full-length polypeptide as translated from a coding open reading frame, or as processed to its mature form, while a polypeptide or peptide informally refers to a degradation fragment or a processing fragment of a protein that nonetheless uniquely or identifiably maps to a particular protein. A polypeptide can be a single linear polymer chain of amino acids bonded together by peptide bonds between the carboxyl and amino groups of adjacent amino acid residues. Polypeptides can be modified, for example, by the addition of carbohydrate, phosphorylation, etc. Proteins can comprise one or more polypeptides.
The term electrophile as used herein refers to a chemical moiety that is capable of accepting an electron pair (e.g. Lewis acid, electron pair acceptor). For example, an electrophile as used herein is a chemical moiety that accepts a pair of electrons thereby forming a covalent bond.
The term “reversible covalent bond” as used herein refers to a labile bond between the amine of a lysine residue and a compound as disclosed herein (e.g. between an electron deficient functional group and the amine of the lysine reside). As used herein a reversible covalent inhibitor or reversible covalent modifier, refer to classes of compounds that comprise a reversible covalent bond. The reversible covalent bond may be a bond as described herein (e.g., carbon nitrogen double bond or sulfur-nitrogen single bond). Bandyopadhyay, A. et al. Curr Opin Chem Biol. October 2016, (34) pp. 110-116; Serafimova, I. M. et al. Nat Chem Biol. May 2012, 8(5), pp. 471-476; and Bradshaw M. J. et al. Nat Chem Biol. Jul. 2015, 11(7): 525-531, describe reversible covalent modifiers or reversible covalent chemical kinetics, the entire contents of each of which are incorporated herein by reference.
The term “contacting” may include allowing two species to react, interact, or physically touch, for example the two species may be an exogenous compound and an AKT1 protein or mutant thereof as described herein.
As used herein “cancer” in a subject refers to the presence of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and certain characteristic morphological features. In some circumstances, cancers will be in the form of a tumor, or such cells that may exist locally within a subject, or circulate in the blood stream as independent cells.
The term “tumor” can refer to a solid or fluid-filled lesion or structure that may be formed by cancerous or non-cancerous cells, such as cells exhibiting aberrant cell growth or division. The terms “mass” and “nodule” are often used synonymously with “tumor”. Tumors include malignant tumors or benign tumors. An example of a malignant tumor can be a carcinoma which is known to comprise transformed cells.
The term “in vivo” is used to describe an event that takes place in a subject's body.
The term “ex vivo” is used to describe an event that takes place outside of a subject's body. An “ex vivo” assay is not performed on a subject. Rather, it is performed upon a sample separate from a subject. An example of an ‘ex vivo’ assay performed on a sample is an ‘in vitro’ assay.
The term “in vitro” is used to describe an event that takes places contained in a container for holding laboratory reagent such that it is separated from the living biological source organism from which the material is obtained. In vitro assays can encompass cell-based assays in which cells alive or dead are employed. In vitro assays can also encompass a cell-free assay in which no intact cells are employed.
The phrase “pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
The term “salt” or “pharmaceutically acceptable salt” refers to salts derived from a variety of organic and inorganic counter ions well known in the art. Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Those of ordinary skill in the art will recognize from this disclosure and the knowledge in the art that further pharmaceutically acceptable salts include those listed by Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, PA, p. 1418 (1985). In general, a pharmaceutically acceptable acid or base salt can be synthesized from a parent compound that contains a basic or acidic moiety by any conventional chemical method. Briefly, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in an appropriate solvent.
The phrase “pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.
The terms “subject,” “individual,” and “patient” may be used interchangeably and refer to humans, as well as non-human mammals (e.g., non-human primates, canines, equines, felines, porcines, bovines, ungulates, lagomorphs, and the like). In various embodiments, the subject can be a human (e.g., adult male, adult female, adolescent male, adolescent female, male child, female child) under the care of a physician or other health worker in a hospital, as an outpatient, or other clinical context. In certain embodiments, the subject may not be under the care or prescription of a physician or other health worker.
As used herein, the phrase “a subject in need thereof” refers to a subject, as described infra, that suffers from, or is at risk for, a pathology to be prophylactically or therapeutically treated with a compound or salt described herein.
The terms “administer”, “administered”, “administers” and “administering” are defined as providing a composition to a subject via a route known in the art, including but not limited to intravenous, intraarterial, oral, parenteral, buccal, topical, transdermal, rectal, intramuscular, subcutaneous, intraosseous, transmucosal, or intraperitoneal routes of administration. In certain embodiments, oral routes of administering a composition can be used. The terms ““administer”, “administered”, “administers” and “administering” a compound should be understood to mean providing a compound of the invention or a prodrug of a compound of the invention to the individual in need.
The terms “determining”, “measuring”, “evaluating”, “assessing, ““assaying, “and “analyzing” are often used interchangeably herein to refer to forms of measurement, and include determining if an element is present or not (for example, detection). These terms can include quantitative, qualitative or quantitative and qualitative determinations. Assessing is alternatively relative or absolute. “Detecting the presence of” includes determining the amount of something present, as well as determining whether it is present or absent.
As used herein, “treatment” or “treating” refers to an approach for obtaining beneficial or desired results with respect to a disease, disorder, or medical condition including, but not limited to, a therapeutic benefit and/or a prophylactic benefit. In certain embodiments, treatment or treating involves administering a compound or composition disclosed herein to a subject. A therapeutic benefit may include the eradication or amelioration of the underlying disorder being treated. Also, a therapeutic benefit may be achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder, such as observing an improvement in the subject, notwithstanding that the subject may still be afflicted with the underlying disorder. In certain embodiments, for prophylactic benefit, the compositions are administered to a subject at risk of developing a particular disease, or to a subject reporting one or more of the physiological symptoms of a disease, even though a diagnosis of this disease may not have been made. Treating can include, for example, reducing, delaying or alleviating the severity of one or more symptoms of the disease or condition, or it can include reducing the frequency with which symptoms of a disease, defect, disorder, or adverse condition, and the like, are experienced by a patient. Treating can be used herein to refer to a method that results in some level of treatment or amelioration of the disease or condition, and can contemplate a range of results directed to that end, including but not restricted to prevention of the condition entirely.
In certain embodiments, the term “prevent” or “preventing” as related to a disease or disorder may refer to a compound that, in a statistical sample, reduces the occurrence of the disorder or condition in the treated sample relative to an untreated control sample, or delays the onset or reduces the severity of one or more symptoms of the disorder or condition relative to the untreated control sample.
A “therapeutic effect,” as that term is used herein, encompasses a therapeutic benefit and/or a prophylactic benefit as described above. A prophylactic effect includes delaying or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or thereof.
The term “kinase activity” as herein refers to a variance in the nucleotide sequence of agene that results in a modulated kinase activity (increased or decreased). The modulated kinase activity is a result of the variance in the nucleic acid and is associated with the protein for which the gene encodes.
“Alkyl” refers to a straight or branched hydrocarbon chain monovalent radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, and preferably having from one to twelve carbon atoms (i.e., C1-12 alkyl). The alkyl is attached to the remainder of the molecule through a single bond. An alkyl chain may be optionally substituted by one or more substituents such as those substituents described herein. In certain embodiments, an alkyl comprises one to twelve carbon atoms (i.e., C1-12 alkyl). In certain embodiments, an alkyl comprises one to eight carbon atoms (i.e., C1-8 alkyl). In other embodiments, an alkyl comprises one to five carbon atoms (i.e., C1-5 alkyl). In other embodiments, an alkyl comprises one to four carbon atoms (i.e., C1-4 alkyl). In other embodiments, an alkyl comprises one to three carbon atoms (i.e., C1-3 alkyl). In other embodiments, an alkyl comprises one to two carbon atoms (i.e., C1-2 alkyl). In other embodiments, an alkyl comprises one carbon atom (i.e., C1 alkyl). In other embodiments, an alkyl comprises five to fifteen carbon atoms (i.e., C5-15 alkyl). In other embodiments, an alkyl comprises five to eight carbon atoms (i.e., C5-8 alkyl). In other embodiments, an alkyl comprises two to five carbon atoms (i.e., C2-5 alkyl). In other embodiments, an alkyl comprises three to five carbon atoms (i.e., C3-5 alkyl). For example, the alkyl group may be attached to the rest of the molecule by a single bond, such as, methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (iso-propyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (iso-butyl), 1,1-dimethylethyl (tert-butyl), 1-pentyl (n-pentyl), and the like.
“Alkenyl” refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon double bond, and preferably having from two to twelve carbon atoms (i.e., C2-12 alkenyl). An alkenyl chain may be optionally substituted by one or more substituents such as those substituents described herein. In certain embodiments, an alkenyl comprises two to eight carbon atoms (i.e., C2-8 alkenyl). In certain embodiments, an alkenyl comprises two to six carbon atoms (i.e., C2-6 alkenyl). In other embodiments, an alkenyl comprises two to four carbon atoms (i.e., C2-4 alkenyl). The alkenyl is attached to the rest of the molecule by a single bond, for example, ethenyl (i.e., vinyl), prop-1-enyl (i.e., allyl), but-1-enyl, pent-1-enyl, penta-1,4-dienyl, and the like.
“Alkynyl” refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one carbon—carbon triple bond, and preferably having from two to twelve carbon atoms (i.e., C2-12 alkynyl). An alkylnyl chain may be optionally substituted by one or more substituents such as those substituents described herein. In certain embodiments, an alkynyl comprises two to eight carbon atoms (i.e., C2-8 alkynyl). In other embodiments, an alkynyl comprises two to six carbon atoms (i.e., C2-6 alkynyl). In other embodiments, an alkynyl comprises two to four carbon atoms (i.e., C2-4 alkynyl). The alkynyl is attached to the rest of the molecule by a single bond, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like.
“Alkylene” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing no unsaturation, and preferably having from one to twelve carbon atoms, for example, methylene, ethylene, propylene, (methyl)ethylene, butylene, and the like. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. An alkylene chain may be optionally substituted by one or more substituents such as those substituents described herein. In certain embodiments, an alkylene comprises one to ten carbon atoms (i.e., C1-10 alkylene). In certain embodiments, an alkylene comprises one to eight carbon atoms (i.e., C1-8 alkylene). In other embodiments, an alkylene comprises one to five carbon atoms (i.e., C1-5 alkylene). In other embodiments, an alkylene comprises one to four carbon atoms (i.e., C1-4 alkylene). In other embodiments, an alkylene comprises one to three carbon atoms (i.e., C1-3 alkylene). In other embodiments, an alkylene comprises one to two carbon atoms (i.e., C1-2 alkylene). In other embodiments, an alkylene comprises one carbon atom (i.e., C1 alkylene). In other embodiments, an alkylene comprises five to eight carbon atoms (i.e., C5-8 alkylene). In other embodiments, an alkylene comprises two to five carbon atoms (i.e., C2-5 alkylene). In other embodiments, an alkylene comprises three to five carbon atoms (i.e., C3-5 alkylene).
“Alkenylene” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing at least one carbon-carbon double bond, and preferably having from two to twelve carbon atoms. The alkenylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. An alkenylene chain may be optionally substituted by one or more substituents such as those substituents described herein. In certain embodiments, an alkenylene comprises two to ten carbon atoms (i.e., C2-10 alkenylene). In certain embodiments, an alkenylene comprises two to eight carbon atoms (i.e., C2-8 alkenylene). In other embodiments, an alkenylene comprises two to five carbon atoms (i.e., C2-5 alkenylene). In other embodiments, an alkenylene comprises two to four carbon atoms (i.e., C2-4 alkenylene). In other embodiments, an alkenylene comprises two to three carbon atoms (i.e., C2-3 alkenylene). In other embodiments, an alkenylene comprises two carbon atoms (i.e., C2 alkenylene). In other embodiments, an alkenylene comprises five to eight carbon atoms (i.e., C5-8 alkenylene). In other embodiments, an alkenylene comprises three to five carbon atoms (i.e., C3-5 alkenylene).
“Alkynylene” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing at least one carbon-carbon triple bond, and preferably having from two to twelve carbon atoms. The alkynylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. An alkynylene chain may be optionally substituted by one or more substituents such as those substituents described herein. In certain embodiments, an alkynylene comprises two to ten carbon atoms (i.e., C2-10 alkynylene). In certain embodiments, an alkynylene comprises two to eight carbon atoms (i.e., C2-8 alkynylene). In other embodiments, an alkynylene comprises two to five carbon atoms (i.e., C2-5 alkynylene). In other embodiments, an alkynylene comprises two to four carbon atoms (i.e., C2-4 alkynylene). In other embodiments, an alkynylene comprises two to three carbon atoms (i.e., C2-3 alkynylene). In other embodiments, an alkynylene comprises two carbon atoms (i.e., C2 alkynylene). In other embodiments, an alkynylene comprises five to eight carbon atoms (i.e., C5-8 alkynylene). In other embodiments, an alkynylene comprises three to five carbon atoms (i.e., C3-5 alkynylene).
The term “Cx-y” when used in conjunction with a chemical moiety, such as alkyl, alkenyl, or alkynyl is meant to include groups that contain from x to y carbons in the chain. For example, the term “C1-6 alkyl” refers to saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups that contain from 1 to 6 carbons. The term -Cx-y alkylene-refers to a alkylene chain with from x to y carbons in the alkylene chain. For example, —C1-6 alkylene-may be selected from methylene, ethylene, propylene, butylene, pentylene, and hexylene, any one of which may be optionally substituted.
The terms “Cx-y alkenyl” and “Cx-y alkynyl” refer to unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond, respectively. The term -Cx-y alkenylene-refers to a alkenylene chain with from x to y carbons in the alkenylene chain. For example, —C2-6 alkenylene-may be selected from ethenylene, propenylene, butenylene, pentenylene, and hexenylene, any one of which may be optionally substituted. An alkenylene chain may have one double bond or more than one double bond in the alkenylene chain. The term -Cx-y alkynylene-refers to a alkynylene chain with from x to y carbons in the alkynylene chain. For example, —C2-6 alkynylene-may be selected from ethynylene, propynylene, butynylene, pentynylene, and hexynylene, any one of which may be optionally substituted. An alkynylene chain may have one triple bond or more than one triple bond in the alkynylene chain.
The term “carbocycle” as used herein refers to a saturated, unsaturated or aromatic ring in which each atom of the ring is carbon. Carbocycle includes 3- to 10-membered monocyclic rings and polycyclic rings (e.g., 6- to 12-membered bicyclic rings). Each ring of a polycyclic carbocycle may be selected from saturated, unsaturated, and aromatic rings. Polycyclic carbocycles may be fused, bridged or spiro-ring systems. Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated, and aromatic rings. Bicyclic carbocycles may be fused, bridged or spiro-ring systems. In some embodiments, the carbocycle is an aryl. In some embodiments, the carbocycle is a cycloalkyl. In some embodiments, the carbocycle is a cycloalkenyl. In an exemplary embodiment, an aromatic ring, e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits, are included in the definition of carbocyclic. Exemplary carbocycles include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl. Carbocycle may be optionally substituted by one or more substituents such as those substituents described herein.
The term “carbocyclene” as used herein refers to a divalent saturated, unsaturated or aromatic ring in which each atom of the ring is carbon. The carbocyclene is attached to the rest of the molecule through a single bond and to the radical group through a single bond. A carbocyclene may be optionally substituted by one or more substituents such as those substituents described herein. Carbocyclene includes divalent 3- to 10-membered monocyclic rings and divalent polycyclic rings (e.g., 6- to 12-membered bicyclic rings). Each ring of a polycyclic carbocyclene may be selected from saturated, unsaturated, and aromatic rings. Polycyclic carbocyclenes may be fused, bridged or spiro-ring systems. Polycyclic carbocyclenes may be fused, bridged or spiro-ring systems. The single bond connecting the carbocyclene to the rest of the molecule and the single bond connecting the carbocyclene to the radical group may be located on the same ring or different rings of a polycyclic carbocyclene. In some embodiments, the carbocycle is an arylene, for example, a phenylene. A “phenylene” as used herein refers to a divalent benzene group. The phenylene is attached to the rest of the molecule through a single bond and to the radical group through a single bond. A phenylene may be optionally substituted by one or more substituents such as those substituents described herein.
“Cycloalkyl” refers to a stable fully saturated monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which includes fused, bridged, or spiro-ring systems, and preferably having from three to twelve carbon atoms (i.e., C3-12 cycloalkyl). In certain embodiments, a cycloalkyl comprises three to ten carbon atoms (i.e., C3-10 cycloalkyl). In other embodiments, a cycloalkyl comprises five to seven carbon atoms (i.e., C5-7 cycloalkyl). The cycloalkyl may be attached to the rest of the molecule by a single bond. Examples of monocyclic cycloalkyls include, e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl radicals include, for example, adamantyl, norbomyl (i.e., bicyclo[2.2.1]heptanyl), norbornenyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Cycloalkyl may be optionally substituted by one or more substituents such as those substituents described herein.
“Cycloalkenyl” refers to a stable unsaturated non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which includes fused or bridged ring systems, preferably having from three to twelve carbon atoms and comprising at least one double bond (i.e., C3-12 cycloalkenyl). In certain embodiments, a cycloalkenyl comprises three to ten carbon atoms (i.e., C3-10 cycloalkenyl). In other embodiments, a cycloalkenyl comprises five to seven carbon atoms (i.e., C5-7 cycloalkenyl). The cycloalkenyl may be attached to the rest of the molecule by a single bond. Examples of monocyclic cycloalkenyls include, e.g., cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. Cycloalkenyl may be optionally substituted by one or more substituents such as those substituents described herein.
“Aryl” refers to a radical derived from an aromatic monocyclic or aromatic polycyclic hydrocarbon ring system by removing a hydrogen atom from a ring carbon atom. The aromatic monocyclic or aromatic multicyclic hydrocarbon ring system contains only hydrogen and carbon and from five to eighteen carbon atoms, where at least one of the rings in the ring system is aromatic, i.e., it contains a cyclic, delocalized (4n+2) p-electron system in accordance with the Huckel theory. The ring system from which aryl groups are derived include, but are not limited to, groups such as benzene, fluorene, indane, indene, tetralin and naphthalene. Aryl may be optionally substituted by one or more substituents such as those substituents described herein.
A “Cx-y carbocycle” is meant to include groups that contain from x to y carbons in a ring. For example, the term “C3-6 carbocycle” can be a saturated, unsaturated or aromatic ring system that contains from 3 to 6 carbon atoms—any one of which may be optionally substituted as provided herein.
The term “heterocycle” as used herein refers to a saturated, unsaturated, non-aromatic or aromatic ring comprising one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycles include 3- to 10-membered monocyclic rings and polycyclic rings (e.g., 6- to 12-membered bicyclic rings). Polycyclic heterocycles may be fused, bridged or spiro-ring systems. Each ring of a polycyclic heterocycle may be selected from saturated, unsaturated, and aromatic rings. In some embodiments, the heterocycle comprises at least one heteroatom selected from oxygen, nitrogen, sulfur, or any combination thereof. In some embodiments, the heterocycle comprises at least one heteroatom selected from oxygen, nitrogen, or any combination thereof. In some embodiments, the heterocycle comprises at least one heteroatom selected from oxygen, sulfur, or any combination thereof. In some embodiments, the heterocycle comprises at least one heteroatom selected from nitrogen, sulfur, or any combination thereof. The heterocycle may be attached to the rest of the molecule through any atom of the heterocycle, valence permitting, such as a carbon or nitrogen atom of the heterocycle. In some embodiments, the heterocycle is a heteroaryl. In some embodiments, the heterocycle is a heterocycloalkyl. Exemplary heterocycles include pyrrolidinyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, piperidinyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, thiophenyl, oxazolyl, thiazolyl, morpholinyl, indazolyl, indolyl, and quinolinyl. Heterocycle may be optionally substituted by one or more substituents such as those substituents described herein. Bicyclic heterocycles may be fused, bridged or spiro-ring systems. In an exemplary embodiment, a heterocycle, e.g., pyridyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Heterocycle may be optionally substituted by one or more substituents such as those substituents described herein.
The term “heterocyclene” as used herein refers to a divalent saturated, unsaturated, non-aromatic or aromatic ring comprising one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. The heterocyclene is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The single bond attaching the heterocyclene group to the rest of the molecule and the single bond attaching the heterocyclene group to the radical group may be each independently connected through any atom of the heterocyclene as valency permits, including a carbon atom in the heterocyclene ring or a heteroatom in the heterocyclene ring. A heterocyclene may be optionally substituted by one or more substituents such as those substituents described herein. Heterocyclenes include 3- to 10-membered monocyclic rings and polycyclic rings (e.g., 6- to 12-membered bicyclic rings). Each ring of a polycyclic heterocyclene may be selected from saturated, unsaturated, and aromatic rings. Polycyclic heterocyclenes may be fused, bridged or spiro-ring systems. The single bond connecting the heterocyclene to the rest of the molecule and the single bond connecting the heterocyclene to the radical group may be located on the same ring or different rings of a polycyclic heterocyclene, and may be attached to the rest of the molecule or the radical group through any atom of the heterocyclene, valence permitting, such as a carbon or nitrogen atom of the heterocycle. In some embodiments, the heterocyclene comprises at least one heteroatom selected from oxygen, nitrogen, sulfur, or any combination thereof. In some embodiments, the heterocyclene comprises at least one heteroatom selected from oxygen, nitrogen, or any combination thereof. In some embodiments, the heterocyclene comprises at least one heteroatom selected from oxygen, sulfur, or any combination thereof. In some embodiments, the heterocyclene comprises at least one heteroatom selected from nitrogen, sulfur, or any combination thereof. In some embodiments, the heterocyclene is a heteroarylene. In some embodiments, the heterocyclene is a heterocycloalkylene.
“Heterocycloalkyl” refers to a stable 3 to 12 membered non-aromatic ring radical that comprises two to twelve carbon atoms and at least one heteroatom wherein each heteroatom may be selected from N, O, Si, P, B, and S atoms. In some embodiments, the heterocycloalkyl comprises at least one heteroatom selected from oxygen, nitrogen, sulfur, or any combination thereof. In some embodiments, the heterocycloalkyl comprises at least one heteroatom selected from oxygen, nitrogen, or any combination thereof. In some embodiments, the heterocycloalkyl comprises at least one heteroatom selected from oxygen, sulfur, or any combination thereof. In some embodiments, the heterocycloalkyl comprises at least one heteroatom selected from nitrogen, sulfur, or any combination thereof. The heterocycloalkyl may be selected from monocyclic or bicyclic, and fused, bridged, or spiro-ring systems. The heteroatoms in the heterocycloalkyl radical are optionally oxidized. One or more nitrogen atoms, if present, are optionally quaternized. The heterocycloalkyl radical is partially or fully saturated. The heterocycloalkyl is attached to the rest of the molecule through any atom of the heterocycloalkyl, valence permitting, such as any carbon or nitrogen atoms of the heterocycloalkyl. Examples of heterocycloalkyl radicals include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxothiomorpholinyl, and 1,1-dioxothiomorpholinyl. Heterocycloalkyl may be optionally substituted by one or more substituents such as those substituents described herein.
The term “heteroaryl” refers to a radical derived from a 3- to 12-membered aromatic ring radical that comprises one to eleven carbon atoms and at least one heteroatom wherein each heteroatom may be selected from N, O, and S. In some embodiments, the heteroaryl comprises at least one heteroatom selected from oxygen, nitrogen, sulfur, or any combination thereof. In some embodiments, the heteroaryl comprises at least one heteroatom selected from oxygen, nitrogen, or any combination thereof. In some embodiments, the heteroaryl comprises at least one heteroatom selected from oxygen, sulfur, or any combination thereof. In some embodiments, the heteroaryl comprises at least one heteroatom selected from nitrogen, sulfur, or any combination thereof. As used herein, the heteroaryl ring may be selected from monocyclic or bicyclic and fused or bridged ring systems wherein at least one of the rings in the ring system is aromatic, i.e., it contains a cyclic, delocalized (4n+2) p-electron system in accordance with the Hückel theory. The heteroatom(s) in the heteroaryl radical may be optionally oxidized. One or more nitrogen atoms, if present, are optionally quatemized. The heteroaryl may be attached to the rest of the molecule through any atom of the heteroaryl, valence permitting, such as a carbon or nitrogen atom of the heteroaryl. Heteroaryl includes aromatic single ring structures, preferably 5- to 6-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like. Heteroaryl may be optionally substituted by one or more substituents such as those substituents described herein. Heteroaryl also includes polycyclic ring systems having two or more rings in which two or more atoms are common to two adjoining rings wherein at least one of the rings is heteroaromatic, e.g., the other rings can be aromatic or non-aromatic carbocyclic, or heterocyclic. Heteroaryl may be optionally substituted by one or more substituents such as those substituents described herein.
An “X-membered heterocycle” refers to the number of endocyclic atoms, i.e., X, in the ring. For example, a 5-membered heteroaryl ring or 5-membered aromatic heterocycle has 5 endocyclic atoms, e.g., triazole, oxazole, thiophene, etc.
“Alkoxy” refers to a radical bonded through an oxygen atom of the formula: O-alkyl, where alkyl is an alkyl chain as defined above.
“Halo” or “halogen” refers to halogen substituents such as bromo, chloro, fluoro and iodo substituents.
As used herein, the term “haloalkyl” or “haloalkane” refers to an alkyl radical, as defined above, that is substituted by one or more halogen radicals, for example, trifluoromethyl, dichloromethyl, bromomethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like. In some embodiments, the alkyl part of the fluoroalkyl radical is optionally further substituted. Examples of halogen substituted alkanes (“haloalkanes”) include halomethane (e.g., chloromethane, bromomethane, fluoromethane, iodomethane), di- and trihalomethane (e.g., trichloromethane, tribromomethane, trifluoromethane, triiodomethane), 1-haloethane, 2-haloethane, 1,2-dihaloethane, 1-halopropane, 2-halopropane, 3-halopropane, 1,2-dihalopropane, 1,3-dihalopropane, 2,3-dihalopropane, 1,2,3-trihalopropane, and any other suitable combinations of alkanes (or substituted alkanes) and halogens (e.g., Cl, Br, F, and I). When an alkyl group is substituted with more than one halogen radical, each halogen may be independently selected for example, 1-chloro,2-fluoroethane.
The term “substituted” refers to moieties having substituents replacing a hydrogen on one or more carbons or substitutable heteroatoms, e.g., an NH or NH2 of a compound. Unless specified otherwise (e.g., by using the terms “substituted” or “optionally substituted”, or by the inclusion of an “—R” group), chemical groups described herein are unsubstituted. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, i.e., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. In certain embodiments, substituted refers to moieties having substituents replacing two hydrogen atoms on the same carbon atom, such as substituting the two hydrogen atoms on a single carbon with an oxo, imino or thioxo group. Unless specified otherwise (e.g., by the phrase “two or more substituents come together to form a carbocycle or heterocycle”), two or more substituents on the same or different substituted carbon or substitutable heteroatom will not come together to form a ring structure (e.g., two substituents on an alkyl chain forming a monocyclic ring, or two substituents on a ring system to form a spiro, fused, or bridged polycyclic ring system). As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds.
In some embodiments, substituents may include any substituents described herein, for example: halogen, hydroxy, oxo (═O), thioxo (═S), cyano (—CN), nitro (—NO2), imino (═N—H), oximo (═N—OH), hydrazino (═N—NH2), —Rb—ORa, —Rb—OC(O)Ra, —Rb—OC(O)ORa, —Rb—OC(O)N(Ra)2, —Rb—N(Ra)2, —Rb—C(O)Ra, —Rb—C(O)ORa, —Rb—C(O)N(Ra)2, —Rb—O—Rc—C(O)N(Ra)2, —Rb—N(Ra)C(O)ORa, —Rb—N(Ra)C(O)Ra, —Rb—N(Ra)S(O)tRa (where t is 1 or 2), —Rb—S(O)tRa (where t is 0, 1, or 2), —Rb—S(O)tORa (where t is 1 or 2), —Rb—S(O)tN(Ra)2 (where t is 1 or 2), and —P(O)(Ra)2; and alkyl, alkenyl, alkynyl, aryl, aralkyl, aralkenyl, aralkynyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, and heteroarylalkyl any one of which may be optionally substituted by alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (═O), thioxo (═S), cyano (—CN), nitro (—NO2), imino (═N—H), oximo(═N—OH), hydrazine(═N—NH2), —Rb—ORa, —Rb—OC(O)Ra, —Rb—OC(O)ORa, —Rb—OC(O)N(Ra)2, —Rb—N(Ra)2, —Rb—C(O)Ra, —Rb—C(O)ORa, —Rb—C(O)N(Ra)2, —Rb—O—Rc—C(O)N(Ra)2, —Rb—N(Ra)C(O)ORa, —Rb—N(Ra)C(O)Ra, —Rb—N(Ra)S(O)tRa (where t is 1 or 2), —Rb—S(O)tRa (where t is 0, 1, or 2), —Rb—S(O)ORa (where t is 1 or 2), —Rb—S(O)tN(Ra)2 (where t is 1 or 2), and —P(O)(Ra)2; wherein each Ra is independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl, wherein each Ra, valence permitting, may be optionally substituted with alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (═O), thioxo (═S), cyano (—CN), nitro (—NO2), imino (═N—H), oximo (═N—OH), hydrazine (═N—NH2), —Rb—ORa,
In some embodiments, “substituent group,” as used herein, means a group selected from the following moieties:
A “size-limited substituent” or “size-limited substituent group,” as used herein, means a group selected from all of the substituents described above for a “substituent group,” wherein each substituted or unsubstituted alkyl is a substituted or unsubstituted C1-C20 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 20 membered heteroalkyl, each substituted or unsubstituted alkenyl is a substituted or unsubstituted C1-C20 alkenyl, each substituted or unsubstituted alkynyl is a substituted or unsubstituted C1-C20 alkynyl, each substituted or unsubstituted carbocycle is a substituted or unsubstituted C3-C8 carbocycle, each substituted or unsubstituted cycloalkenyl is a substituted or unsubstituted C3-C8 cycloalkenyl, each substituted or unsubstituted heterocycle is a substituted or unsubstituted 3 to 8 membered heterocycle, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C8 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 8 membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted C6-C10 aryl, and each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5 to 10 membered heteroaryl.
A “lower substituent” or “lower substituent group,” as used herein, means a group selected from all of the substituents described above for a “substituent group,” wherein each substituted or unsubstituted alkyl is a substituted or unsubstituted C1-C8 alkyl, each substituted or unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8 membered heteroalkyl, each substituted or unsubstituted cycloalkyl is a substituted or unsubstituted C3-C7 cycloalkyl, each substituted or unsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 7 membered heterocycloalkyl, each substituted or unsubstituted aryl is a substituted or unsubstituted phenyl, each substituted or unsubstituted alkenyl is a substituted or unsubstituted C1-C8 alkenyl, each substituted or unsubstituted alkynyl is a substituted or unsubstituted C1-C8 alkynyl, each substituted or unsubstituted carbocycle is a substituted or unsubstituted C3-C7 carbocycle, each substituted or unsubstituted cycloalkenyl is a substituted or unsubstituted C3-C7 cycloalkenyl, each substituted or unsubstituted heterocycle is a substituted or unsubstituted 5 to 6 membered heterocycle, and each substituted or unsubstituted heteroaryl is a substituted or unsubstituted 5 to 6 membered heteroaryl.
In some embodiments, each substituted group (e.g. substituted alkyl, alkenyl, alkynyl, heteroalkyl, carbocycle, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycle, heterocycloalkalkylene, alkenylene, alkynylene, heteroalkylene, carbocyclene, arylene, heterarylene and/or heterocyclene) described in the compounds herein is substituted with at least one substituent group. In other embodiments, at least one or all of these groups are substituted with at least one size-limited substituent group. In other embodiments, at least one or all of these groups are substituted with at least one lower substituent group.
While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.
In some aspects, the present disclosure provides an AKT1 protein covalently bound to a compound, wherein the compound is covalently bound to a lysine residue of the AKT1 protein. In some embodiments, the compound is exogenous. In some embodiments, the exogenous compound is selected from an exogenous AKT1 inhibitor and an exogenous AKT1 activator. In some embodiments, the exogenous compound is an exogenous AKT1 modulator. In some embodiments, the exogenous compound is an exogenous AKT1 inhibitor.
In some embodiments, the AKT1 protein is selected from a wild-type AKT1 protein and a mutated AKT1 protein. In some embodiments, the AKT1 protein is a mutated AKT1 protein. In some embodiments, the mutated AKT1 protein comprises a mutation selected from a E17K mutation, a E40K mutation, and a E49K mutation. In some embodiments, the mutated AKT1 protein comprises a E17K mutation. In some embodiments, the mutated AKT1 protein comprises a E40K mutation. In some embodiments, the mutated AKT1 protein comprises a E49K mutation.
In some embodiments, the exogenous compound is in contact a lysine residue of the AKT1 protein as described herein. In some embodiments, the contact is between the lysine reside of the AKT1 protein and the exogenous compound is a covalent bond. In some embodiments, the lysine reside is selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, the lysine residue is K17. In some embodiments, the lysine residue is selected from K158, K163, and K179. In some embodiments, the lysine residue is K158. In some embodiments, the lysine residue is K163. In some embodiments, the lysine residue is K179. In some embodiments, the lysine residue is selected from K40, K49, K276, and K297. In some embodiments, the lysine residue is K40. In some embodiments, the lysine residue is K49. In some embodiments, the lysine residue is K276. In some embodiments, the lysine residue is K297. In some embodiments, the lysine residue is selected from K17 and K297.
In some embodiments, the covalent bond between the exogenous compound and the lysine residue is a reversible covalent bond. In some embodiments, the reversible covalent bond is selected from a single bond, a double bound, and a triple bond. In some embodiments, the reversible covalent bond is a single bond. In some embodiments the reversible covalent bond is a double bond. In some embodiments, the reversible covalent bond is a triple bond. In some embodiments, the reversible covalent bond is a double bond between a carbon atom on the exogenous compound and the nitrogen atom on the sidechain of the lysine residue.
In some embodiments, the covalent bond between the exogenous compound and the lysine residue is a reversible covalent bond, wherein the lysine residue is selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, the covalent bond between the exogenous compound and the lysine residue is a reversible covalent bond selected from a single bond, a double bound, and a triple bond, wherein the lysine residue is selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, the covalent bond between the exogenous compound and the lysine residue is a reversible covalent single bond, wherein the lysine residue is selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, the covalent bond between the exogenous compound and the lysine residue is a reversible covalent double bond, wherein the lysine residue is selected from K17, K40, K49, K158, K163, K179, K276, and K297.
In some embodiments, the covalent bond between the exogenous compound and the lysine residue is a reversible covalent bond, wherein the lysine residue is selected from K17, K40, and K49. In some embodiments, the covalent bond between the exogenous compound and the lysine residue is a reversible covalent bond, wherein the lysine residue is K17. In some embodiments, the covalent bond between the exogenous compound and the lysine residue is a reversible covalent bond selected from a single bond, a double bound, and a triple bond, wherein the lysine residue is K17. In some embodiments, the covalent bond between the exogenous compound and the lysine residue is a reversible covalent single bond, wherein the lysine residue is K17. In some embodiments, the covalent bond between the exogenous compound and the lysine residue is a reversible covalent double bond, wherein the lysine residue is K17.
In some embodiments, the reversible covalent bond in the in vivo AKT1 protein comprises a carbon-nitrogen interaction. In some embodiments, the carbon-nitrogen interaction is selected form a carbon-nitrogen single bond and a carbon-nitrogen double bond. In some embodiments, the carbon-nitrogen interaction is a carbon-nitrogen single bond. In some embodiments, the carbon-nitrogen interaction is a carbon-nitrogen double bond.
In some embodiments, the AKT1 protein is covalently bound with the exogenous compound, wherein the exogenous compound is bound at only one residue of the AKT1 protein. In some embodiments, the AKT1 protein is covalently bond with the exogenous compound via one covalent bond. In some embodiments, the AKT1 protein is covalently bound with the exogenous compound, wherein the exogenous compound is bound at one lysine residue. In some embodiments, the AKT1 protein has a single covalent bond between a lysine residue and the exogenous compound. In some embodiments, the AKT1 protein has a single covalent bond between K17 and the exogenous compound. In some embodiments, the AKT1 protein has a single covalent bond between K158 and the exogenous compound. In some embodiments, the AKT1 protein has a single covalent bond between K163 and the exogenous compound. In some embodiments, the AKT1 protein has a single covalent bond between K179 and the exogenous compound.
In some embodiments, the exogenous compound has reduced engagement at other lysine residues when covalently bound at a lysine residue selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, the exogenous compound is in contact with one lysine residue selected from K17, K40, K49, K158, K163, K179, K276, and K297, and has reduced engagement at the remaining lysine residues. In some embodiments, the exogenous compound is in contact with K17, and has reduced engagement at K40, K49, K158, K163, K179, K276, or K297. In some embodiments, the exogenous compound is in contact with K158, and has reduced engagement at K17, K40, K49, K163, K179, K276, or K297. In some embodiments, the exogenous compound is in contact with K163, and has reduced engagement at K17, K40, K49, K158, K179, K276, or K297. In some embodiments, the exogenous compound is in contact with K179, and has reduced engagement at K17, K40, K49, K158, K163, K276, or K297. In some embodiments, the exogenous compound is in contact with K40, and has reduced engagement at K17, K49, K158, K163, K179, K276, or K297. In some embodiments, the exogenous compound is in contact with K49, and has reduced engagement at K17, K40, K158, K163, K179, K276, or K297. In some embodiments, the exogenous compound is in contact with K276, and has reduced engagement at K17, K40, K49, K158, K163, K179, or K297. In some embodiments, the exogenous compound is in contact with K297, and has reduced engagement at K17, K40, K49, K158, K163, K179, or K276.
In some embodiments, the AKT1 protein has a single lysine residue covalently bound with the exogenous compound. In some embodiments, the AKT1 protein has a single lysine residue covalently bound with the exogenous compound, wherein the single lysine residue is selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, the AKT1 protein has a single lysine residue covalently bound with the exogenous compound, wherein the single lysine residue is K17. In some embodiments, the AKT1 protein has a single lysine residue covalently bound with the exogenous compound, wherein the single lysine residue is K158. In some embodiments, the AKT1 protein has a single lysine residue covalently bound with the exogenous compound, wherein the single lysine residue is K163. In some embodiments, the AKT1 protein has a single lysine residue covalently bound with the exogenous compound, wherein the single lysine residue is K179. In some embodiments, the AKT1 protein has a single lysine residue covalently bound with the exogenous compound, wherein the single lysine residue is K40. In some embodiments, the AKT1 protein has a single lysine residue covalently bound with the exogenous compound, wherein the single lysine residue is K49. In some embodiments, the AKT1 protein has a single lysine residue covalently bound with the exogenous compound, wherein the single lysine residue is K276. In some embodiments, the AKT1 protein has a single lysine residue covalently bound with the exogenous compound, wherein the single lysine residue is K297.
In some embodiments, the AKT1 protein is in vivo. In some embodiments, the AKT1 protein is in vitro. In some embodiments, the AKT1 protein is ex vivo. In some embodiments, the AKT1 protein is an in vivo engineered protein.
In some embodiments, the AKT1 protein is an in vivo engineered AKT1 protein, wherein the in vivo engineered AKT1 protein is generated by contacting the AKT1 protein in vivo with the exogenous compound. In some embodiments, the AKT1 protein is a mammalian in vivo engineered AKT1 protein, wherein the in vivo engineered AKT1 protein is generated by contacting the AKT1 protein in vivo with the exogenous compound. In some embodiments, the AKT1 protein is a human in vivo engineered AKT1 protein, wherein the in vivo engineered AKT1 protein is generated by contacting the AKT1 protein in vivo with the exogenous compound.
In some embodiments, the reversible covalent bond in the in vivo AKT1 protein is between a carbon atom and a nitrogen atom. In some embodiments, the reversible covalent bond in the in vivo AKT1 protein is between a carbon atom of the exogenous compound and a nitrogen atom of the lysine residue. In some embodiments, the reversible covalent bond in the in vivo AKT1 protein is between a carbon atom of the exogenous compound and a nitrogen atom of the lysine residue selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, the reversible covalent bond in the in vivo AKT1 protein is between a carbon atom of the exogenous compound and a nitrogen atom of the K17 lysine residue. In some embodiments, the reversible covalent bond in the in vivo AKT1 protein is between a carbon atom of the exogenous compound and a nitrogen atom of the K158 lysine residue. In some embodiments, the reversible covalent bond in the in vivo AKT1 protein is between a carbon atom of the exogenous compound and a nitrogen atom of the K163 lysine residue. In some embodiments, the reversible covalent bond in the in vivo AKT1 protein is between a carbon atom of the exogenous compound and a nitrogen atom of the K179 lysine residue.
In some embodiments, the reversible covalent bond in the in vivo AKT1 protein is a carbon-nitrogen double bond. In some embodiments, the reversible covalent bond in the in vivo AKT1 protein is a carbon-nitrogen double bond between the exogenous compound and the lysine residue. In some embodiments, the carbon-nitrogen double bond is between the exogenous compound and the lysine residue, wherein the lysine residue is selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, the carbon-nitrogen double bond is between the exogenous compound and the K17 lysine residue. In some embodiments, the carbon-nitrogen double bond is between the exogenous compound and the K158 lysine residue. In some embodiments, the carbon-nitrogen double bond is between the exogenous compound and the K163 lysine residue. In some embodiments, the carbon-nitrogen double bond is between the exogenous compound and the K179 lysine residue. In some embodiments, the carbon-nitrogen double bond is between the exogenous compound and the K40 lysine residue. In some embodiments, the carbon-nitrogen double bond is between the exogenous compound and the K49 lysine residue. In some embodiments, the carbon-nitrogen double bond is between the exogenous compound and the K276 lysine residue. In some embodiments, the carbon-nitrogen double bond is between the exogenous compound and the K297 lysine residue.
In some embodiments, the carbon-nitrogen double bond is an imine bound. In some embodiments, the imine bond is between a carbon atom of the exogenous compound and a nitrogen atom of the lysine residue In some embodiments, the imine bond is between the exogenous compound and the lysine residue, wherein the lysine residue is selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, the imine bond is between the exogenous compound and the K 17 lysine residue. In some embodiments, the imine bond is between the exogenous compound and the K158 lysine residue. In some embodiments, the imine bond is between the exogenous compound and the K163 lysine residue. In some embodiments, the imine bond is between the exogenous compound and the K179 lysine residue. In some embodiments, the imine bond is between the exogenous compound and the K40 lysine residue. In some embodiments, the imine bond is between the exogenous compound and the K49 lysine residue. In some embodiments, the imine bond is between the exogenous compound and the K276 lysine residue. In some embodiments, the imine bond is between the exogenous compound and the K297 lysine residue.
In some embodiments, the exogenous compound comprises a functional group. In some embodiments, the exogenous compound comprises an aldehyde functional group. In some embodiments, the reversible covalent bond is between the aldehyde functional group and the lysine residue. In some embodiments, the reversible covalent bond is between the aldehyde functional group and the lysine residue selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, the reversible covalent bond is between the aldehyde functional group and the K17 lysine residue. In some embodiments, the reversible covalent bond is between the aldehyde functional group and the K158 lysine residue. In some embodiments, the reversible covalent bond is between the aldehyde functional group and the K163 lysine residue. In some embodiments, the reversible covalent bond is between the aldehyde functional group and the K179 lysine residue. In some embodiments, the reversible covalent bond is between the aldehyde functional group and the K40 lysine residue. In some embodiments, the reversible covalent bond is between the aldehyde functional group and the K49 lysine residue. In some embodiments, the reversible covalent bond is between the aldehyde functional group and the K276 lysine residue. In some embodiments, the reversible covalent bond is between the aldehyde functional group and the K297 lysine residue.
In some embodiments, the carbon-nitrogen bond is a reversible bond that results from a reversible reaction. In some embodiments, the carbon-nitrogen bond is a reversible bond that results from a reversible reaction between the exogenous compound and a lysine residue. In some embodiments, the carbon-nitrogen bond is a reversible bond that results from a reversible reaction between the exogenous compound and a lysine residue selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, the carbon-nitrogen bond is a reversible bond that results from a reversible reaction between the exogenous compound and the K17 lysine residue. In some embodiments, the carbon-nitrogen bond is a reversible bond that results from a reversible reaction between the exogenous compound and the K158 lysine residue. In some embodiments, the carbon-nitrogen bond is a reversible bond that results from a reversible reaction between the exogenous compound and the K163 lysine residue. In some embodiments, the carbon-nitrogen bond is a reversible bond that results from a reversible reaction between the exogenous compound and the K179 lysine residue. In some embodiments, the carbon-nitrogen bond is a reversible bond that results from a reversible reaction between the exogenous compound and the K40 lysine residue. In some embodiments, the carbon-nitrogen bond is a reversible bond that results from a reversible reaction between the exogenous compound and the K49 lysine residue. In some embodiments, the carbon-nitrogen bond is a reversible bond that results from a reversible reaction between the exogenous compound and the K276 lysine residue. In some embodiments, the carbon-nitrogen bond is a reversible bond that results from a reversible reaction between the exogenous compound and the K297 lysine residue.
In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of a lysine residue and an aldehyde functional group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of a lysine residue and an aldehyde functional group on the exogenous compound, wherein the lysine residue is selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K17 and an aldehyde functional group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K158 and an aldehyde functional group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K163 and an aldehyde functional group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K179 and an aldehyde functional group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K40 and an aldehyde functional group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K49 and an aldehyde functional group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K276 and an aldehyde functional group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K297 and an aldehyde functional group on the exogenous compound.
In some embodiments, the aldehyde functional group of the exogenous compound is an aromatic aldehyde. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of a lysine residue and the aromatic aldehyde group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of a lysine residue and the aromatic aldehyde group on the exogenous compound, wherein the lysine residue is selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K17 and the aromatic aldehyde group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K158 and the aromatic aldehyde group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K163 and the aromatic aldehyde group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K179 and the aromatic aldehyde group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K40 and the aromatic aldehyde group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K49 and the aromatic aldehyde group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K276 and the aromatic aldehyde group on the exogenous compound. In some embodiments, the carbon-nitrogen double bond results from a reversible reaction between the amine functional group of K297 and the aromatic aldehyde group on the exogenous compound.
In some embodiments, the AKT1 modulator is selected from N-(4-(2-(2-aminopyridin-3-yl)-5-(3-ethynylphenyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)-4-formyl-3-hydroxybenzamide (Compound A) and 4-(2-((4-(2-(2-aminopyridin-3-yl)-5-(3-ethynylphenyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)amino)ethyl)-2-hydroxybenzaldehyde (Compound B). In some embodiments, the AKT1 modulator is selected from
In some embodiments, the AKT1 modulator is Compound A. In some embodiments, the AKT1 modulator is Compound B.
In some embodiments, the AKT1 modulator is a compound or salt as disclosed herein. In some embodiments, the AKT1 modulator is selected from a compound or salt of Formula (I) or Formula (II). In some embodiments, the AKT1 modulator is selected from a compound or salt in Table 1.
In some embodiments, the AKT1 inhibitor is selected from N-(4-(2-(2-aminopyridin-3-yl)-5-(3-ethynylphenyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)-4-formyl-3-hydroxybenzamide (Compound A) and 4-(2-((4-(2-(2-aminopyridin-3-yl)-5-(3-ethynylphenyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)amino)ethyl)-2-hydroxybenzaldehyde (Compound B). In some embodiments, the AKT1 inhibitor is selected from
In some embodiments, the AKT1 inhibitor is Compound A. In some embodiments, the AKT1 inhibitor is Compound B.
In some embodiments, the AKT1 inhibitor is a compound or salt as disclosed herein. In some embodiments, the AKT1 inhibitor is selected from a compound or salt of Formula (I) or Formula (II). In some embodiments, the AKT1 inhibitor is selected from a compound or salt in Table 1.
In some embodiments, the exogenous compound is selected from N-(4-(2-(2-aminopyridin-3-yl)-5-(3-ethynylphenyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)-4-formyl-3-hydroxybenzamide (Compound A) and 4-(2-((4-(2-(2-aminopyridin-3-yl)-5-(3-ethynylphenyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)amino)ethyl)-2-hydroxybenzaldehyde (Compound B). In some embodiments, the exogenous compound is selected from
In some embodiments, the exogenous compound is Compound A. In some embodiments, the exogenous compound is Compound B.
In some embodiments, the exogenous compound is a compound or salt as disclosed herein. In some embodiments, the exogenous compound is selected from a compound or salt of Formula (I) or Formula (II). In some embodiments, the exogenous compound is selected from a compound or salt in Table 1.
In certain aspects the present disclosure provides an in vivo engineered AKT1 protein comprising a non-naturally occurring reversible covalent modification at a lysine residue, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue. In some embodiments, the in vivo engineered AKT1 protein is a human in vivo engineered AKT1 protein
In another aspect, the present disclosure provides an in vivo engineered AKT1 protein comprising a non-naturally occurring reversible covalent modification at a lysine residue selected from K17, K40, K49, K158, K163, K179, K276, and K297, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue. In some embodiments, the in vivo engineered AKT1 protein is a human in vivo engineered AKT1 protein
In another aspect, the present disclosure provides an in vivo engineered AKT1 protein comprising a non-naturally occurring reversible covalent modification at a lysine residue selected from K17, K40, K49, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue. In some embodiments, the in vivo engineered AKT1 protein is a human in vivo engineered AKT1 protein.
In another aspect, the present disclosure provides an in vivo engineered AKT1 protein comprising a non-naturally occurring reversible covalent modification at a K17 lysine residue, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the K17 lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue. In some embodiments, the in vivo engineered AKT1 protein is a human in vivo engineered AKT1 protein
In another aspect, the present disclosure provides an in vivo engineered AKT1 protein comprising a non-naturally occurring reversible covalent modification at a lysine residue selected from K158, K163, and K179, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue. In some embodiments, the in vivo engineered AKT1 protein is a human in vivo engineered AKT1 protein.
In another aspect, the present disclosure provides an in vivo engineered AKT1 protein comprising a non-naturally occurring reversible covalent modification at a lysine residue selected from K40, K49, K276 and K297, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue. In some embodiments, the in vivo engineered AKT1 protein is a human in vivo engineered AKT1 protein
In certain aspects the present disclosure provides a human in vivo engineered AKT1 protein comprising a non-naturally occurring reversible covalent modification at a lysine residue selected from K17, K40, K49, K158, K163, K179, K276, and K297, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue.
In another aspect the present disclosure provides a human in vivo engineered AKT1 protein comprising a non-naturally occurring reversible covalent modification at a lysine residue selected from K17, K40, and K49, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue.
In another aspect, the present disclosure provides a human in vivo engineered AKT1 protein comprising a non-naturally occurring reversible covalent modification at a K17 lysine residue, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the K17 lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue.
In another aspect, the present disclosure provides a human in vivo engineered AKT1 protein comprising a non-naturally occurring reversible covalent modification at a lysine residue selected from K158, K163, and K179, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue.
In another aspect, the present disclosure provides a human in vivo engineered AKT1 protein comprising a non-naturally occurring reversible covalent modification at a lysine residue selected from K40, K49, K276, and K297, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue.
In certain aspects the present disclosure provides an in vivo engineered AKT1 protein comprises a mutation selected from E17K, E40K, and E49K, a non-naturally occurring reversible covalent modification at a lysine residue, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue.
In certain aspects the present disclosure provides an in vivo engineered AKT1 protein comprising a E17K mutation, a non-naturally occurring reversible covalent modification at a lysine residue, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue.
In another aspect, the present disclosure provides an in vivo engineered AKT1 protein comprising a E17K mutation, a non-naturally occurring reversible covalent modification at a lysine residue selected from K17, K40, K49, K158, K163, K179, K276, and K297, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue. In some embodiments, the in vivo engineered AKT1 protein is a human in vivo engineered AKT1 protein.
In certain aspects the present disclosure provides a human in vivo engineered AKT1 protein comprising a E17K mutation, a non-naturally occurring reversible covalent modification at a lysine residue, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue.
In another aspect, the present disclosure provides a human in vivo engineered AKT1 protein comprising a E17K mutation, a non-naturally occurring reversible covalent modification at a lysine residue, the reversible covalent modification being generated from an in vivo nucleophilic reaction between an exogenous aromatic aldehyde and the lysine residue of AKT1, wherein the exogenous aromatic aldehyde undergoes a nucleophilic addition with the amine functional group on the lysine residue and forming a carbon-nitrogen double bond between the exogenous aromatic aldehyde and the amine functional group on the lysine residue.
In some aspects the present disclosure provides a method of modifying an AKT1 protein as disclosed herein. In some embodiments, the method of covalently modifying an AKT1 protein, comprises contacting the AKT1 protein with an exogenous compound, wherein the exogenous compound comprises a reversible electrophilic moiety thereby forming a reversible covalent AKT1 adduct. In some embodiments, the contacting is in vitro or in vivo. In some embodiments, the contacting is in vitro. In some embodiments, the contacting is in vivo. In some embodiments, the AKT1 protein is wild type AKT1 or a mutated AKT1. In some embodiments, the mutated AKT1 is E17K AKT1. In some embodiments, the wild type AKT1 protein is wild type. In some embodiments, the AKT1 protein is E17K AKT1. In some embodiments, the exogenous compound is an AKT1 inhibitor. In some embodiments, the reversible covalent moiety on the AKT1 inhibitor is an aromatic aldehyde. In some embodiments, the reversible covalent AKT1 adduct is formed between the reversible covalent moiety and a lysine reside of the AKT1 protein. In some embodiments, the reversible covalent AKT1 adduct is formed between the aromatic aldehyde and the lysine residue of the AKT1 protein. In some embodiments, reversible covalent AKT1 adduct is formed between the reversible covalent moiety and a lysine residue of the AKT1 protein selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, reversible covalent AKT1 adduct is formed between the aromatic aldehyde and a lysine residue of the AKT1 protein selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, reversible covalent AKT1 adduct is formed between the aromatic aldehyde and the K17 lysine residue of the AKT1 protein.
In another aspect, the method of covalently modifying an AKT1 protein, comprises contacting the AKT1 protein with an exogenous AKT1 modulator, wherein the AKT1 modulator comprises a reversible electrophilic moiety thereby forming a reversible covalent AKT1 adduct. In some embodiments, the contacting is in vitro or in vivo. In some embodiments, the contacting is in vitro. In some embodiments, the contacting is in vivo. In some embodiments, the AKT1 protein is wild type AKT1 or a mutated AKT1. In some embodiments, the mutated AKT1 is selected from E17K AKT1, E40K AKT1, and E49K AKT1. In some embodiments, the mutated AKT1 is E17K AKT1. In some embodiments, the wild type AKT1 protein is wild type. In some embodiments, the AKT1 protein is E17K AKT1. In some embodiments, the reversible covalent moiety on the AKT1 modulator is an aromatic aldehyde. In some embodiments, the reversible covalent AKT1 adduct is formed between the reversible covalent moiety and a lysine reside of the AKT1 protein. In some embodiments, the reversible covalent AKT1 adduct is formed between the aromatic aldehyde and the lysine residue of the AKT1 protein. In some embodiments, reversible covalent AKT1 adduct is formed between the reversible covalent moiety and a lysine residue of the AKT1 protein selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, reversible covalent AKT1 adduct is formed between the aromatic aldehyde and a lysine residue of the AKT1 protein selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, reversible covalent AKT1 adduct is formed between the aromatic aldehyde and the K17 lysine residue of the AKT1 protein. In some embodiments, the exogenous AKT1 modulator is an AKT1 inhibitor.
In certain aspects the present disclosure provides a method of attenuating AKT1 activity In some embodiments, the method of covalently modifying an AKT1 protein, comprises contacting the AKT1 protein with an exogenous AKT1 inhibitor, wherein the AKT1 modulator comprises a reversible electrophilic moiety thereby forming a reversible covalent AKT1 adduct. In some embodiments, the contacting is in vitro or in vivo. In some embodiments, the contacting is in vitro. In some embodiments, the contacting is in vivo. In some embodiments, the AKT1 protein is wild type AKT1 or a mutated AKT1. In some embodiments, the mutated AKT1 is selected from E17K AKT1, E40K AKT1, and E49K AKT1. In some embodiments, the mutated AKT1 is E17K AKT1. In some embodiments, the wild type AKT1 protein is wild type. In some embodiments, the AKT1 protein is E17K AKT1. In some embodiments, the reversible covalent moiety on the AKT1 inhibitor is an aromatic aldehyde. In some embodiments, the reversible covalent AKT1 adduct is formed between the reversible covalent moiety and a lysine reside of the AKT1 protein. In some embodiments, the reversible covalent AKT1 adduct is formed between the aromatic aldehyde and the lysine residue of the AKT1 protein. In some embodiments, reversible covalent AKT1 adduct is formed between the reversible covalent moiety and a lysine residue of the AKT1 protein selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, reversible covalent AKT1 adduct is formed between the aromatic aldehyde and a lysine residue of the AKT1 protein selected from K17, K40, K49, K158, K163, K179, K276, and K297. In some embodiments, reversible covalent AKT1 adduct is formed between the aromatic aldehyde and the K17 lysine residue of the AKT1 protein.
In some aspects, the method of attenuating AKT1 activity, comprises contacting AKT1 protein with an exogenous compound, wherein the exogenous compound comprises a reversible electrophilic moiety. In some embodiments, the AKT1 protein is wild type AKT1 or a mutated AKT1. In some embodiments, the mutated AKT1 is selected from E17K AKT1, E40K AKT1, and E49K AKT1. In some embodiments, the mutated AKT1 is E17K AKT1. In some embodiments, the wild type AKT1 protein is wild type. In some embodiments, the contacting is in vitro or in vivo. In some embodiments, the contacting is in vitro.
In some embodiments, following the contacting, the AKT1 activity is attenuated by 50% to 95% relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by 75% to 95% relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by 50% or more relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by 70% or more relative to a control in the absence of the exogenous compound.
In some embodiments, following the contacting, the AKT1 activity is attenuated by about 50% to about 95% relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by about 75% to about 95% relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by about 50% or more relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by about 70% or more relative to a control in the absence of the exogenous compound.
In some embodiments, following the contacting, the AKT1 activity is attenuated by at least 50% to at least 95% relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by at least 75% to at least 95% relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by at least 50% relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by at least 70% relative to a control in the absence of the exogenous compound.
In some embodiments, following the contacting, the AKT1 activity is attenuated by at most 50% to at most 95% relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by at most 75% to at most 95% relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by at most 50% relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by at most 70% relative to a control in the absence of the exogenous compound. In some embodiments, following the contacting, the AKT1 activity is attenuated by at most 50%, at most 55%, at most 60%, at most 65%, at most 70%, at most 75%, at most 80%, at most 85%, at most 90%, or at most 95% relative to a control in the absence of the exogenous compound.
In some embodiments, the exogenous compound is more selective toward mutated AKT1 than wild-type AKT1. In some embodiments, the mutated AKT1 is E17K AKT1. In some embodiments, the exogenous compound is 2-fold to 100-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is 2-fold to 10-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is 2-fold to 5-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is 2-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is 3-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is 4-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is 5-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is 10-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 25-fold, 50-fold, 75-fold, or 100-fold more selective for E17K AKT1 over wild-type AKT1.
In some embodiments, the exogenous compound is at least 2-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is at least 3-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is at least 4-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is at least 5-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is at least 10-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is at least 2-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, at least 75-fold, or at least 100-fold more selective for E17K AKT1 over wild-type AKT1.
In some embodiments, the exogenous compound is about 2-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is about 3-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is about 4-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is about 5-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is about 10-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, about 10-fold, about 15-fold, about 20-fold, about 25-fold, about 50-fold, about 75-fold, or about 100-fold more selective for E17K AKT1 over wild-type AKT1.
In some embodiments, the exogenous compound is at most 2-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is at most 3-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is at most 4-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is at most 5-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is at most 10-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is at most 2-fold more selective for E17K AKT1 over wild-type AKT1. In some embodiments, the exogenous compound is at most 2-fold, at most 3-fold, at most 4-fold, at most 5-fold, at most 6-fold, at most 7-fold, at most 8-fold at most 9-fold, at most 10-fold, at most 15-fold, at most 20-fold, at most 25-fold, at most 50-fold at most 75-fold, or at most 100-fold more selective for E17K AKT1 over wild-type AKT1.
In another aspect, the present disclosure provides a method of attenuating AKT1 activity, comprising contacting AKT1 protein with an AKT1 inhibitor, wherein the AKT1 inhibitor comprises a reversible electrophilic moiety. In some embodiments, the AKT1 protein is wild type AKT1 or a mutated AKT1. In some embodiments, the mutated AKT1 is selected from E17K AKT1, E40K AKT1, and E49K AKT1. In some embodiments, the mutated AKT1 is E17K AKT1. In some embodiments, the wild type AKT1 protein is wild type. In some embodiments, the contacting is in vitro or in vivo. In some embodiments, the contacting is in vitro.
In some embodiments, following the contacting, the AKT1 activity is attenuated by 50% to 95% relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by 75% to 95% relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by 50% or more relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by 70% or more relative to a control in the absence of the exogenous AKT1 inhibitor.
In some embodiments, following the contacting, the AKT1 activity is attenuated by about 50% to about 95% relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by about 75% to about 95% relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by about 50% or more relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by about 70% or more relative to a control in the absence of the exogenous AKT1 inhibitor.
In some embodiments, following the contacting, the AKT1 activity is attenuated by at least 50% to at least 95% relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by at least 75% to at least 95% relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by at least 50% relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by at least 70% relative to a control in the absence of the exogenous AKT1 inhibitor.
In some embodiments, following the contacting, the AKT1 activity is attenuated by at most 50% to at most 95% relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by at most 75% to at most 95% relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by at most 50% relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by at most 70% relative to a control in the absence of the exogenous AKT1 inhibitor. In some embodiments, following the contacting, the AKT1 activity is attenuated by at most 50%, at most 55%, at most 60%, at most 65%, at most 70%, at most 75%, at most 80%, at most 85%, at most 90%, or at most 95% relative to a control in the absence of the exogenous AKT1 inhibitor.
A compound of Formula (I):
In some embodiments, for the compound or salt of Formula (I), R0 is independently selected at each occurrence from hydrogen, alkyl, heteroalkyl, alkenyl, and alkynyl, any of which is independently unsubstituted or substituted. In some embodiments, R0 is independently selected at each occurrence from hydrogen, alkyl, and heteroalkyl, any of which is independently unsubstituted or substituted. R0 is hydrogen.
In some embodiments, for the compound or salt of Formula (I), R1 is selected from hydrogen, halogen, alkyl, heteroalkyl, alkenyl, alkynyl, —OR10, —SR10, —N(R10)2, —NO2, and —CN, wherein each alkyl, heteroalkyl, alkenyl, and alkynyl is independently unsubstituted or substituted. In some embodiments, R1 is selected from hydrogen, halogen, alkyl, heteroalkyl, alkenyl, and alkynyl, wherein each alkyl, wherein each alkyl, heteroalkyl, alkenyl, and alkynyl is independently unsubstituted or substituted. In some embodiments, R1 is selected from hydrogen, halogen, alkyl, and heteroalkyl, wherein each alkyl and heteroalkyl is independently unsubstituted or substituted In some embodiments, R1 is selected from hydrogen, halogen, alkyl, heteroalkyl, —OR10, —SR10, —N(R10)2, —NO2, and —CN, wherein each alkyl, and heteroalkyl is independently unsubstituted or substituted; and wherein R10 is independently selected at each occurrence from hydrogen, unsubstituted alkyl, and substituted alkyl. In some embodiments, R1 is selected from hydrogen, halogen, unsubstituted alkyl, —OR10, —SR10, —N(R10)2, —NO2, and —CN, and wherein R10 is independently selected at each occurrence from hydrogen, unsubstituted alkyl, and substituted alkyl. In some embodiments, R1 is selected from hydrogen, halogen, and unsubstituted alkyl. In some embodiments, R1 is hydrogen.
In some embodiments, for the compound or salt of Formula (I), R2 is selected from hydrogen, halogen, alkyl, heteroalkyl, alkenyl, alkynyl, —OR10, —SR10, —N(R10)2, —NO2, and —CN, wherein each alkyl, heteroalkyl, alkenyl, and alkynyl is independently unsubstituted or substituted. In some embodiments, R2 is selected from hydrogen, halogen, alkyl, heteroalkyl, alkenyl, and alkynyl, wherein each alkyl, wherein each alkyl, heteroalkyl, alkenyl, and alkynyl is independently unsubstituted or substituted. In some embodiments, R2 is selected from hydrogen, halogen, alkyl, and heteroalkyl, wherein each alkyl and heteroalkyl is independently unsubstituted or substituted In some embodiments, R2 is selected from hydrogen, halogen, alkyl, heteroalkyl, —OR10, —SR10, —N(R10)2, —NO2, and —CN, wherein each alkyl, and heteroalkyl is independently unsubstituted or substituted; and wherein R10 is independently selected at each occurrence from hydrogen, unsubstituted alkyl, and substituted alkyl. In some embodiments, R2 is selected from hydrogen, halogen, unsubstituted alkyl, —OR10, —SR10, —N(R10)2, —NO2, and —CN, and wherein R10 is independently selected at each occurrence from hydrogen, unsubstituted alkyl, and substituted alkyl. In some embodiments, R2 is selected from hydrogen, halogen, and unsubstituted alkyl.
In some embodiments, for the compound or salt of Formula (I), n is selected from 0, 1, 2, and 3. In some embodiments, n is selected from 0, 1, and 2. In some embodiments, n is selected from 0 and 1. In some embodiments, n is 0.
In some embodiments, the compound or salt of Formula (I) is a compound represented by the structure of Formula (I-A):
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), A1 and A2 are each independently selected from: hydrogen, halogen, —OR11, —SR11, —N(R11)2, —C(O)R11, —C(O)N(R11)2, —N(R11)C(O)R11, —NO2, —CN, alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle of A1 and A2 is independently unsubstituted or substituted. In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), A1 and A2 are each independently selected from: hydrogen, halogen, —OR11, —SR11, —N(R11)2, —C(O)R11, —C(O)N(R11)2, —N(R11)C(O)R11, —NO2, —CN, alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle of A1 and A2 is independently unsubstituted or substituted; and R11 is independently selected at each occurrence from hydrogen, alkyl, heteroalkyl, carbocycle, and heterocycle, any of which is independently unsubstituted or substituted. In some embodiments, A1 and A2 are each independently selected from: hydrogen, halogen, —OR11, —N(R11)2, —C(O)R11, —C(O)N(R11)2, —N(R11)C(O)R11, alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle of A1 and A2 is independently unsubstituted or substituted. In some embodiments, A1 and A2 are each independently selected from: hydrogen, halogen, —OR11, —N(R11)2, alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle of A1 and A2 is independently unsubstituted or substituted. In some embodiments, A1 and A2 are each independently selected from: hydrogen, halogen, —OR11, —N(R11)2, alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle of A1 and A2 is independently unsubstituted or substituted; and R11 is independently selected at each occurrence from hydrogen and unsubstituted alkyl. In some embodiments, A1 and A2 are each independently selected from: alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle, any of which is independently unsubstituted or substituted. In some embodiments, A1 and A2 are each independently selected from: alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle, any of which is independently unsubstituted or substituted. In some embodiments, A1 and A2 are hydrogen.
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), A1 and A2 are each independently selected from: hydrogen, halogen, —OR11, —SR11, —N(R11)2, —C(O)R11, —C(O)N(R11)2, —N(R11)C(O)R11, —NO2, —CN, alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle of A1 and A2 is independently unsubstituted or substituted with one or more substituents selected from
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), A1 and A2 are each independently selected from: hydrogen, halogen, —OR11, —SR11, —N(R11)2, —C(O)R11, —C(O)N(R11)2, —N(R11)C(O)R11, —NO2, —CN, alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle of A1 and A2 is independently unsubstituted or substituted with one or more substituents selected from
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), A1 and A2 are each independently selected from: hydrogen, halogen, —OR11, —N(R11)2, —C(O)R11, —C(O)N(R11)2, —N(R11)C(O)R11, alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle of A and A2 is independently unsubstituted or substituted with one or more substituents selected from
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), A1 and A2 are each independently selected from: hydrogen, halogen, —OR11, —N(R11)2, alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle of A1 and A2 is independently unsubstituted or substituted with one or more substituents selected from
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), A1 and A2 are each independently selected from: hydrogen, halogen, —OR11, —N(R11)2, alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle of A1 and A2 is independently unsubstituted or substituted with one or more substituents selected from
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), A1 and A2 are each independently selected from: hydrogen, halogen, —OR11, —N(R11)2, alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle of A1 and A2 is independently unsubstituted or substituted with one or more substituents selected from
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), A1 is selected from: hydrogen, halogen, —OR11, —SR11, —N(R11)2, —C(O)R11, —C(O)N(R11)2, —N(R11)C(O)R11, —NO2, —CN, alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle is independently unsubstituted or substituted; and wherein R11 is independently selected at each occurrence from hydrogen and unsubstituted alkyl. In some embodiments, A1 is selected from: hydrogen, halogen, alkyl, carbocycle and heterocycle, wherein each alkyl, carbocycle and heterocycle is independently unsubstituted or substituted. In some embodiments, A1 is selected from: hydrogen and phenyl. In some embodiments, A1 is hydrogen.
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), A2 is selected from: hydrogen, halogen, —OR11, —SR11, —N(R11)2, —C(O)R11, —C(O)N(R11)2, —N(R11)C(O)R11, —NO2, —CN, alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle is independently unsubstituted or substituted. In some embodiments, A2 is selected from: hydrogen, halogen, —OR11, —N(R11)2, —C(O)R11, —C(O)N(R11)2, —N(R11)C(O)R11, alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle is independently unsubstituted or substituted. In some embodiments, A2 is selected from: hydrogen, halogen, —OR11, —N(R11)2, alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle is independently unsubstituted or substituted. In some embodiments, A2 is selected from: hydrogen, halogen, —OR11, —N(R11)2, alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle is independently unsubstituted or substituted; and wherein R11 is independently selected at each occurrence from hydrogen and unsubstituted alkyl. In some embodiments, A2 is selected from: alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle, any of which is independently unsubstituted or substituted. In some embodiments, A2 is selected from: alkyl, heteroalkyl, alkynyl, carbocycle and heterocycle, any of which is independently unsubstituted or substituted.
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), A2 is selected from: hydrogen, halogen, —OR11, —SR11, —N(R11)2, —C(O)R11, —C(O)N(R11)2, —N(R11)C(O)R11, —NO2, —CN, alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle, wherein each alkyl, heteroalkyl, alkenyl, alkynyl, carbocycle and heterocycle is independently unsubstituted or substituted with one or more substituents selected from
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), A2 is selected from: hydrogen, halogen, —OR11, —SR11, —N(R11)2, —C(O)R11, —C(O)N(R11)2,
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), R20 is selected from heterocyclene and phenylene, any of which is unsubstituted or substituted. In some embodiments, R20 is selected from unsubstituted heterocyclene and unsubstituted phenylene. In some embodiments, R20 is selected from 5- to 6-membered heterocyclene and phenylene, any of which are unsubstituted or substituted. In some embodiments, R20 is selected from unsubstituted 5- to 6-membered heterocyclene and unsubstituted phenylene. In some embodiments, R20 is selected from 6-membered heteroarylene and phenylene, any of which are unsubstituted or substituted. In some embodiments, R20 is selected from unsubstituted 6-membered heteroarylene and unsubstituted phenylene. In some embodiments, R20 is selected from pyridinylene and phenylene, any one of which are substituted or unsubstituted. In some embodiments, R20 is selected from unsubstituted pyridinylene and unsubstituted phenylene. In some embodiments, R20 is unsubstituted phenylene.
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L is a bond or represented by -L1-L2-L3-L4-.
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L1, L2, L3, and L4 are each independently selected from (a) and (b):
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L1, L2, L3, and L4 are each independently selected from (a) and (b):
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L1, L2, L3, and L4 are each independently selected from (a) and (b):
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L1, L2, L3, and L4 are each independently selected from (a) and (b):
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L1, L2, L3, and L4 are each independently selected from (a) and (b):
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L2, L3, and L4 are absent; and L1 is selected from
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L2, L3, and L4 are absent; and L1 is selected from
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L2, L3, and L4 are absent; and L1 is selected from
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L2, L3, and L4 are absent; and L1 is selected from
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L2, L3, and L4 are absent; and L1 is selected from alkylene, heteroalkylene, alkynylene, carbocyclene, and heterocyclene, any of which is independently unsubstituted or substituted.
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L2, L3, and L4 are absent; and L1 is selected from unsubstituted alkylene, substituted alkylene, unsubstituted heteroalkylene, and substituted heteroalkylene.
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L3 and L4 are absent; and L1 and L2 are independently selected from:
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L3 and L4 are absent; and L1 and L2 are independently selected from:
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L3 and L4 are absent; and L1 and L2 are independently selected from:
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L3 and L4 are absent; and L1 and L2 are independently selected from:
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L3 and L4 are absent; and L1 and L2 are independently selected from:
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L3 and L4 are absent; and L1 and L2 are independently selected from:
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L4 is absent; and each of L1, L2, and L3 are independently selected from:
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L4 is absent; and each of L1, L2, and L3 are independently selected from:
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L4 is absent; and each of L1, L2, and L3 are independently selected from:
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L4 is absent; and each of L1, L2, and L3 are independently selected from:
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L4 is absent; and each of L1, L2, and L3 are independently selected from:
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), L4 is absent; and each of L1, L2, and L3 are independently selected from:
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), R21 is selected from heterocycle and carbocycle, any of which is substituted with C(O)H, wherein R21 is optionally further substituted. In some embodiments, R21 is selected from 5- to 6-membered heterocycle and C3-6 carbocycle, any of which is substituted with C(O)H, wherein R21 is optionally further substituted. In some embodiments, R21 is selected from 6-membered heteroaryl and phenyl, any of which is substituted with C(O)H, wherein R21 is optionally further substituted. In some embodiments, R21 is selected from pyridinyl and phenyl, any of which is substituted with C(O)H, wherein R21 is optionally further substituted. In some embodiments, R21 is phenyl substituted with C(O)H and is optionally further substituted.
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), R21 is selected from heterocycle and carbocycle, any of which is substituted with C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from: halogen, unsubstituted alkyl, unsubstituted
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), R21 is selected from 5- to 6-membered heterocycle and C3-6 carbocycle, any of which is substituted with C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from: halogen, unsubstituted alkyl, unsubstituted haloalkyl, —OR15, —SR15, —N(R15)2, —B(OR15)2, —C(O)R15, —C(O)N(R15)2, —C(O)OR15, —OC(O)R15, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15)C(O)N(R15)2, —S(O)R15, —S(O)2R15, —NO2, ═O, ═S, ═N(R15), and —CN. In some embodiments, R21 is selected from 5- to 6-membered heterocycle and C3-6 carbocycle, any of which is substituted with C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from: halogen, unsubstituted alkyl, unsubstituted haloalkyl, —OR15, —SR15, —N(R15)2, —B(OR15)2, —C(O)R15, —C(O)N(R15)2, —C(O)OR15, —OC(O)R15, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15)C(O)N(R15)2, —S(O)R15, —S(O) 2R15, —NO2, ═O, ═S, ═N(R15), and —CN; and wherein R15 is independently selected at each occurrence from hydrogen, unsubstituted alkyl, unsubstituted carbocycle, unsubstituted heterocycle, and substituted heterocycle. In some embodiments, R21 is selected from 5- to 6-membered heterocycle and C3-6 carbocycle, any of which is substituted with C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from: halogen, unsubstituted alkyl, unsubstituted haloalkyl, —OR15, —C(O)R15, —C(O)N(R15)2, —C(O)OR15, —OC(O)R15, —N(R15)C(O)R15, —N(R15)C(O)N(R15)2, ═O, and —CN. In some embodiments, R21 is selected from 5- to 6-membered heterocycle and C3-6 carbocycle, any of which is substituted with C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from: halogen, unsubstituted alkyl, unsubstituted haloalkyl, —OR15, —N(R15)C(O)R15, and —N(R15)C(O)N(R15)2. In some embodiments, R21 is selected from 5- to 6-membered heterocycle and C3-6 carbocycle, any of which is substituted with C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from: halogen, unsubstituted alkyl, unsubstituted haloalkyl, —OR15, —N(R15)C(O)R15, and —N(R15)C(O)N(R15)2; and R15 is independently selected at each occurrence from hydrogen, unsubstituted alkyl, unsubstituted carbocycle, unsubstituted heterocycle, and substituted heterocycle.
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), R21 is selected from heteroaryl and phenyl, any of which is substituted with C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from: halogen, unsubstituted alkyl, unsubstituted haloalkyl, —OR15, —SR15, —N(R15)2, —B(OR15)2, —C(O)R15, —C(O)N(R15)2, —C(O)OR15, —OC(O)R15, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15)C(O)N(R15)2, —S(O)R15, —S(O)2R15, —NO2, ═O, ═S, ═N(R15), and —CN. In some embodiments, R21 is selected from heteroaryl and phenyl, any of which is substituted with C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from: halogen, unsubstituted alkyl, unsubstituted haloalkyl, —OR15, —SR15, —N(R15)2, —B(OR15)2, —C(O)R15, —C(O)N(R15)2, —C(O)OR15, —OC(O)R15, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15)C(O)N(R15)2, —S(O)R15, —S(O)2R15, —NO2, ═O, ═S, ═N(R15), and —CN; and wherein R15 is independently selected at each occurrence from hydrogen, unsubstituted alkyl, unsubstituted carbocycle, unsubstituted heterocycle, and substituted heterocycle. In some embodiments, R21 is selected from heteroaryl and phenyl, any of which is substituted with C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from: halogen, unsubstituted alkyl, unsubstituted haloalkyl, —OR15, —C(O)R15, —C(O)N(R15)2, —C(O)OR15, —OC(O)R15, —N(R15)C(O)R15, —N(R15)C(O)N(R15)2, ═O, and —CN. In some embodiments, R21 is selected from heteroaryl and phenyl, any of which is substituted with C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from: halogen, unsubstituted alkyl, unsubstituted haloalkyl, —OR15, —N(R15)C(O)R15, and —N(R15)C(O)N(R15)2. In some embodiments, R21 is selected from heteroaryl and phenyl, any of which is substituted with C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from: halogen, unsubstituted alkyl, unsubstituted haloalkyl, —OR15, —N(R15)C(O)R15 and —N(R15)C(O)N(R15)2; and R15 is independently selected at each occurrence from hydrogen, unsubstituted alkyl, unsubstituted carbocycle, unsubstituted heterocycle, and substituted heterocycle.
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), R21 is selected from pyridinyl and phenyl, any of which is substituted with C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from: halogen, unsubstituted alkyl, unsubstituted
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), R10, R11, and R14 are each independently selected at each occurrence from hydrogen, alkyl, heteroalkyl, carbocycle, and heterocycle, any of which is independently unsubstituted or substituted. In some embodiments, R10, R11, and R14 are each independently selected at each occurrence from hydrogen, unsubstituted alkyl, and substituted alkyl. In some embodiments, R10, R11, and R14 are each hydrogen.
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), R10 is independently selected at each occurrence from hydrogen, alkyl, heteroalkyl, carbocycle, and heterocycle, any of which is independently unsubstituted or substituted. In some embodiments, R10 is independently selected at each occurrence from hydrogen, unsubstituted alkyl, and substituted alkyl. In some embodiments, R10 is hydrogen.
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), R11 is independently selected at each occurrence from hydrogen, alkyl, heteroalkyl, carbocycle, and heterocycle, any of which is independently unsubstituted or substituted. In some embodiments, R11 is independently selected at each occurrence from hydrogen, unsubstituted alkyl, and substituted alkyl. In some embodiments, R11 is hydrogen.
In some embodiments, for the compound or salt of Formula (I) or Formula (I-A), R14, is independently selected at each occurrence from hydrogen, alkyl, heteroalkyl, carbocycle, and heterocycle, any of which is independently unsubstituted or substituted. In some embodiments, R14 is independently selected at each occurrence from hydrogen, unsubstituted alkyl, and substituted alkyl. In some embodiments, R14 is hydrogen.
In one aspect, the present disclosure provides a compound represented by the structure of Formula (II):
wherein,
In some embodiments, for the compound or salt of Formula (II), R1 is selected from hydrogen, halogen, C1-4 alkyl, C1-4 haloalkyl, —OR10, —SR10, —N(R10)2, —NO2, and —CN. In some embodiments, R1 is selected from hydrogen, halogen, C1-4 alkyl, and C1-4 haloalkyl. In some embodiments, R1 is hydrogen.
In some embodiments, for the compound or salt of Formula (II), R2 is selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR10, —SR10, —N(R10)2, —NO2, and —CN. In some embodiments, R2 is selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR10, —SR10, —N(R10)2, —NO2, and —CN; and R10 is independently selected at each occurrence from hydrogen and C1-4 alkyl. In some embodiments, R2 is selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR10, —SR10, —N(R10)2, —NO2, and —CN; and R10 is hydrogen. In some embodiments, R2 is selected from halogen, C1-4 alkyl, and C1-4 haloalkyl.
In some embodiments, for the compound or salt of Formula (II), n is selected from 0, 1, 2, and 3. In some embodiments, n is selected from 0, 1, and 2. In some embodiments, n is selected from 0 and 1. In some embodiments, n is 0.
In some embodiments, the compound or salt of Formula (II) is a compound represented by the structure of Formula (II-A):
In some embodiments, for the compound or salt of Formula (II) or (II-A), A1 and A2 are each independently selected from:
In some embodiments, for the compound or salt of Formula (II) or (II-A), A1 and A2 are each independently selected from:
In some embodiments, for the compound or salt of Formula (II) or (II-A), A1 and A2 are each independently selected from:
In some embodiments, for the compound or salt of Formula (II) or (II-A), A1 and A2 are each independently selected from:
In some embodiments, for the compound or salt of Formula (II) or (II-A), A1 and A2 are each independently selected from:
In some embodiments, for the compound or salt Formula (II) or (II-A), A1 and A2 are each independently selected from:
In some embodiments, for the compound or salt Formula (II) or (II-A), A1 and A2 are each independently selected from:
In some embodiments, for the compound or salt of Formula (II) or (II-A), A1 and A2 are each independently selected from:
In some embodiments, for the compound or salt of Formula (II) or (II-A), A1 and A2 are each independently selected from:
In some embodiments, for the compound or salt of Formula (II) or (II-A), A1 and A2 are each independently selected from hydrogen, halogen, —OR11, —N(R11)2, C1-6 alkyl, and C2-6 alkynyl, wherein the C1-6 alkyl, and C2-6 alkynyl are each optionally substituted with one or more substituents independently selected from halogen and —OR11. In some embodiments, A1 and A2 are hydrogen.
In some embodiments, for the compound or salt of Formula (II) or (II-A), A1 is independently selected from:
In some embodiments, for the compound or salt of Formula (II) or (II-A), A1 is independently selected from:
In some embodiments, for the compound or salt of Formula (II) or (II-A), A1 is selected from hydrogen, halogen, 3- to 10-membered heterocycle, and C3-10 carbocycle. In some embodiments, A1 is selected from hydrogen and C3-10 carbocycle. In some embodiments, A1 is selected from hydrogen and C3-6 carbocycle. In some embodiments, A1 is selected from hydrogen and phenyl. In some embodiments, A1 is hydrogen.
In some embodiments, the compound or salt of Formula (II) is a compound represented by the structure of Formula (II-B):
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from:
In some embodiments, for the compound or salt Formula (II), (II-A), or (II-B), A2 is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from hydrogen, halogen, —OR11, —N(R11)2, C1-6 alkyl, and C2-6 alkynyl, wherein the C1-6 alkyl, and C2-6 alkynyl are each optionally substituted with one or more substituents independently selected from halogen and —OR11. In some embodiments, A2 is selected from hydrogen, halogen, —OR11, —N(R11)2, C1-6 alkyl, and C2-6 alkynyl, wherein the C1-6 alkyl, and C2-6 alkynyl are each optionally substituted with one or more substituents independently selected from halogen and —OR11; and R11 is independently selected at each occurrence from hydrogen, C1-4 alkyl, C3-s carbocycle, and 3- to 8-membered heterocycle, wherein the C3-8 carbocycle and 3- to 8-membered heterocycle are each optionally substituted with one or more C1-4 alkyl. In some embodiments, In some embodiments, A2 is hydrogen.
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from hydrogen, fluoro, chloro,
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from 3- to 10-membered heterocycle and C3-10 carbocycle, any one of which is optionally substituted with one or more substituents independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from 3- to 10-membered heterocycle and C3-10 carbocycle, any one of which is optionally substituted with one or more substituents independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from 3- to 10-membered heterocycle and C3-10 carbocycle, any one of which is optionally substituted with one or more substituents independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from 3- to 10-membered heterocycle and C3-10 carbocycle, any one of which is optionally substituted with one or more substituents independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from 3- to 10-membered heterocycle and C3-10 carbocycle, one of which is optionally substituted with one or more substituents independently selected from methyl, fluoro,
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from cyclopropyl, phenyl, pyrazolyl, morpholine, pyridinyl, 8-Oxa-3-azabicyclo[3.2.1]octanyl, 3-Oxa-8-azabicyclo[3.2.1]octanyl any one of which is optionally substituted with one or more substituents independently selected from halogen, C1-3 alkyl, —C(O)N(R11)2, —N(R11)C(O)R11, —N(R11)S(O)2R11, and 3- to 10-membered heterocycle, wherein the 3- to 10-membered heterocycle is optionally substituted with one or more substituents independently selected from —N(R11)C(O)R11 and 5-membered heterocycle, wherein the 5-membered heterocycle is optionally substituted with ═O.
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from cyclopropyl, phenyl, pyrazolyl, morpholine, pyridinyl, 8-Oxa-3-azabicyclo[3.2.1]octanyl, 3-Oxa-8-azabicyclo[3.2.1]octanyl any one of which is optionally substituted with one or more substituents independently selected from halogen, C1-3 alkyl, —N(R11)S(O)2R11, and 3- to 10-membered heterocycle, wherein the 3- to 10-membered heterocycle is optionally substituted with one or more substituents independently selected from —N(R11)C(O)R11 and 5-membered heterocycle, wherein the 5-membered heterocycle is optionally substituted with ═O.
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from cyclopropyl, phenyl, pyrazolyl, morpholine, pyridinyl, 8-Oxa-3-azabicyclo[3.2.1]octanyl, 3-Oxa-8-azabicyclo[3.2.1]octanyl any one of which is optionally substituted with one or more substituents independently selected from halogen, C1-3 alkyl, —C(O)N(R11)2, —N(R11)C(O)R11, —N(R11)S(O)2R11, morpholine, and piperidinyl, wherein the piperidinyl is optionally substituted with one or more substituents independently selected from —N(R11)C(O)R11 and 5-membered heterocycle, wherein the 5-membered heterocycle is optionally substituted with ═O.
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from cyclopropyl, phenyl, pyrazolyl, morpholine, pyridinyl, 8-Oxa-3-azabicyclo[3.2.1]octanyl, 3-Oxa-8-azabicyclo[3.2.1]octanyl any one of which is optionally substituted with one or more substituents independently selected from halogen, C1-3 alkyl, —N(R11)S(O)2R11, morpholine, and piperidinyl, wherein the piperidinyl is optionally substituted with one or more substituents independently selected from —N(R11)C(O)R11 and 5-membered heterocycle, wherein the 5-membered heterocycle is optionally substituted with ═O.
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from cyclopropyl, phenyl, pyrazolyl, morpholine, pyridinyl, 8-Oxa-3-azabicyclo[3.2.1]octanyl, 3-Oxa-8-azabicyclo[3.2.1]octanyl any one of which is optionally substituted with one or more substituents independently selected from: halogen, C1-3 alkyl, —C(O)N(R11)2, —N(R11)C(O)R11, —N(R11)S(O)2R11, morpholine, and piperidinyl, wherein the piperidinyl is optionally substituted with one or more substituents independently selected from —N(R11)C(O)R11 and pyrrolidinone.
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from cyclopropyl, phenyl, pyrazolyl, morpholine, pyridinyl, 8-Oxa-3-azabicyclo[3.2.1]octanyl, 3-Oxa-8-azabicyclo[3.2.1]octanyl any one of which is optionally substituted with one or more substituents independently selected from: halogen, C1-3 alkyl, —N(R11)S(O)2R11, morpholine, and piperidinyl, wherein the piperidinyl is optionally substituted with one or more substituents independently selected from —N(R11)C(O)R11 and pyrrolidinone.
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from cyclopropyl, phenyl, pyrazolyl, morpholine, pyridinyl, 8-Oxa-3-azabicyclo[3.2.1]octanyl, 3-Oxa-8-azabicyclo[3.2.1]octanyl any one of which is optionally substituted with one or more substituents independently selected from: halogen, C1-3 alkyl, —N(R11)S(O)2R11, morpholine, and piperidinyl, wherein the piperidinyl is optionally substituted with one or more substituents independently selected from —N(R11)C(O)R11 and pyrrolidinone; and R11 is independently selected at each occurrence from hydrogen and C1-4 alkyl.
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from cyclopropyl, phenyl, pyrazolyl, morpholine, pyridinyl, 8-Oxa-3-azabicyclo[3.2.1]octanyl, 3-Oxa-8-azabicyclo[3.2.1]octanyl any one of which is optionally substituted with one or more substituents independently selected from methyl, fluoro,
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A) or (II-B) A2 is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), A2 is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), R20 is selected from 5- to 6-membered heterocyclene and phenylene, any one of which is optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR12, —SR12, —N(R12)2, —NO2, ═O, ═S, ═N(R12), and —CN. In some embodiments, R20 is selected from 5- to 6-membered heterocyclene and phenylene, any one of which is optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR12, —SR12,
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), R20 is selected from 5- to 6-membered heterocyclene and phenylene, any one of which is optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, and —CN. In some embodiments, R20 is selected from 5- to 6-membered heterocyclene and phenylene.
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), R20 is selected from 6-membered heteroarylene and phenylene, any one of which is optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR12, —SR12, —N(R12)2, —NO2, and —CN. In some embodiments, R20 is selected from 6-membered heteroarylene and phenylene, any one of which is optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR12, —SR12, —N(R12)2, —NO2, and —CN; and R12 is independently selected at each occurrence from hydrogen and C1-4 alkyl.
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), R20 is selected from 6-membered heteroarylene and phenylene, any one of which is optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, and —CN. In some embodiments, R20 is selected from 6-membered heteroarylene and phenylene.
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), R20 is selected from pyridinylene and phenylene, any one of which is optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR12, —SR12, —N(R12)2, —NO2, and —CN. In some embodiments, R20 is selected from pyridinylene and phenylene, any one of which is optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR12, —SR12, —N(R12)2, —NO2, and —CN; and R12 is independently selected at each occurrence from hydrogen and C1-4 alkyl.
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), R20 is selected from pyridinylene and phenylene, any one of which is optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, and —CN. In some embodiments, R20 is selected from pyridinylene and phenylene. In some embodiments, R20 is phenylene.
In some embodiments, for the compound or salt of Formula (II), (II-A), or (II-B), R20 is selected fro
wherein * represents the attachment to L.
In some embodiments, the compound or salt of Formula (II) is a compound represented by the structure of Formula (II-C):
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L2, L3, and L4 are absent; and L1 is selected from C1-6 alkylene optionally substituted with one or more substituents independently selected from halogen, —OR14, —N(R14)2, and ═O. In some embodiments, L2, L3, and L4 are absent; and L1 is selected from C1-6 alkylene optionally substituted with one or more substituents independently selected from halogen, —OR14, —N(R14)2, and ═O; and R14 is selected from hydrogen.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L2, L3, and L4 are absent; and L1 is selected from C1-3 alkylene optionally substituted with ═O.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L2, L3, and L4 are absent; and L1 is selected from
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and each of L1 and L2 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and each of L1 and L2 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and each of L1 and L2 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and each of L1 and L2 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and each of L1 and L2 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and each of L1 and L2 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and each of L1 and L2 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and each of L1 and L2 are independently selected from C1-4 alkylene, C2-4 alkynylene, and 3- to 6-membered heterocyclene, any one of which is optionally substituted with one or more substituents independently selected from halogen, —OR13, —SR13, —N(R13)2, and —CN. In some embodiments, L3 and L4 are absent; and each of L1 and L2 are independently selected from C1-4 alkylene, C2-4 alkynylene, and 3- to 6-membered heterocyclene, any one of which is optionally substituted with one or more substituents independently selected from halogen, —OR13, —SR13, —N(R13)2, and —CN; and R13 is selected from each occurrence from hydrogen and C1-4 alkyl. In some embodiments, L3 and L4 are absent; and each of L1 and L2 are independently selected from C1-4 alkylene, and piperidinylene.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L3 and L4 are absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L is
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B3), or (II-C), L is
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L is:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and each of L1, L2, and L3 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and each of L1, L2, and L3 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and each of L1, L2, and L3 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and each of L1, L2, and L3 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and each of L1, L2, and L3 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and each of L1, L2, and L3 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and each of L1, L2, and L3 are independently selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L4 is absent; and
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L is selected from:
and In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), L is selected from:
In some embodiments, for the compound of salt of Formula (II), (II-A), (II-B), and (II), L is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from 5- to 6-membered heterocycle and C3-6 carbocycle, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —SR15, —N(R15)2, —B(OR15)2, —C(O)R15, —C(O)N(R15)2, —C(O)OR15, —OC(O)R15, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15)C(O)N(R15)2, —S(O)R15, —S(O)2R15, —NO2, ═O, ═S, ═N(R15), and —CN. In some embodiments, R21 is selected from 5- to 6-membered heterocycle and C3-6 carbocycle, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —SR15, —N(R15)2, —B(OR15)2, —C(O)R15, —C(O)N(R15)2, —C(O)OR15, —OC(O)R15, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15)C(O)N(R15)2, —S(O)R15, —S(O)2R15, —NO2, ═O, ═S, ═N(R15), and —CN; and R15 is independently selected at each occurrence from hydrogen, C1-4 alkyl, C1-4 haloalkyl, C3-8 carbocycle, and 3- to 8-membered heterocycle, wherein the C3-8 carbocycle and 3- to 8-membered heterocycle are each optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, and ═O.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from 5- to 6-membered heterocycle and C3-6 carbocycle, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —N(R15)2, —C(O)R15, —C(O)N(R15)2, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15) C(O)N(R15)2, and —CN.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from 5- to 6-membered heterocycle and C3-6 carbocycle, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —N(R15)C(O)R15, and —N(R15)C(O)N(R15)2. In some embodiments, R21 is selected from 5- to 6-membered heterocycle and C3-6 carbocycle, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —N(R15)C(O)R15, and —N(R15)C(O)N(R15)2; and R15 is independently selected at each occurrence from hydrogen and C1-4 alkyl.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from 5- to 6-membered heterocycle and C3-6 carbocycle, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from fluoro, chloro, bromo, hydroxyl, methyl,
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from 6-membered heteroarylene and phenylene, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR5, —SR5, —N(R15)2, —B(OR15)2, —C(O)R15, —C(O)N(R15)2, —C(O)OR15, —OC(O)R15, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15)C(O)N(R15)2, —S(O)R15, —S(O)2R15, —NO2, and —CN.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from 6-membered heteroarylene and phenylene, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —N(R15)2, —C(O)R15, —C(O)N(R15)2, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15)C(O)N(R15)2, and —CN.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from 6-membered heteroarylene and phenylene, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —N(R15)C(O)R15, and —N(R15)C(O)N(R15)2.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from 6-membered heteroarylene and phenylene, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from fluoro, chloro, bromo, hydroxyl, methyl,
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from pyridinylene and phenylene, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —SR15, —N(R15)2, —B(OR15)2, —C(O)R15, —C(O)N(R15)2, —C(O)OR15, —OC(O)R15, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15)C(O)N(R15)2, —S(O)R15, —S(O)2R15, —NO2, and —CN. In some embodiments, R21 is selected from pyridinylene and phenylene, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —SR15, —N(R15)2, —B(OR15)2, —C(O)R15, —C(O)N(R15)2, —C(O)OR15, —OC(O)R15, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15)C(O)N(R15)2, —S(O)R15, —S(O)2R11, —NO2, and —CN; and R15 is independently selected at each occurrence from hydrogen, C1-4 alkyl, C1-4 haloalkyl, C3-8 carbocycle, and 3- to 8-membered heterocycle, wherein the C3-8 carbocycle and 3- to 8-membered heterocycle are each optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, and ═O.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from pyridinylene and phenylene, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR5, —N(R11)2, —C(O)R15, —C(O)N(R15)2, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15)C(O)N(R15)2, and —CN.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from pyridinylene and phenylene, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —N(R15)C(O)R1′ and —N(R15)C(O)N(R15)2. In some embodiments, R21 is selected from pyridinylene and phenylene, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —N(R15)C(O)R15, and —N(R15)C(O)N(R15)2; and R11 is independently selected at each occurrence from hydrogen and C1-4 alkyl.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from pyridinylene and phenylene, any one of which is substituted with —C(O)H, wherein R21 is further optionally substituted with one or more substituents independently selected from fluoro, chloro, bromo, hydroxyl, methyl,
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from
further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —ORD, —SR11, —N(R15)2, —B(OR15)2, —C(O)R15, —C(O)N(R15)2, —C(O)OR15, —OC(O)R15, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15)C(O)N(R15)2, —S(O)R15, —S(O)2R15, —NO2, and —CN.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from
further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —N(R15)2, —C(O)R15, —C(O)N(R15)2, —N(R15)C(O)R15, —N(R15)S(O)2R15, —N(R15)C(O)N(R15)2, and —CN.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from
further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —N(R15)C(O)R15, and —N(R15)C(O)N(R15)2. In some embodiments, R21 is selected from
further optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, —OR15, —N(R15)C(O)R15, and —N(R15)C(O)N(R15)2; and R15 is independently selected at each occurrence from hydrogen and C1-4 alkyl.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from
further optionally substituted with one or more substituents independently selected from fluoro, chloro, bromo, methyl,
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R21 is selected from:
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R10, R11, R12, R13, R14, and R15 are each independently selected at each occurrence from hydrogen, C1-4 alkyl, C1-4 haloalkyl, C3-8 carbocycle, and 3- to 8-membered heterocycle, wherein the C3-8 carbocycle and 3- to 8-membered heterocycle are each optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, and ═O. In some embodiments, R10, R11, R12, R13, R14, and R15 are each independently selected at each occurrence from hydrogen, C1-4 alkyl, C3-8 carbocycle, and 3- to 8-membered heterocycle, wherein the C3-8 carbocycle and 3- to 8-membered heterocycle are each optionally substituted with one or more C1-4 alkyl. In some embodiments, R10, R, R12, R13, R14, and R15 are each independently selected at each occurrence from hydrogen and C1-4 alkyl. In some embodiments, R10, R, R12, R13, R14, and R15 are hydrogen.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R10 is independently selected at each occurrence from hydrogen, C1-4 alkyl, C1-4 haloalkyl, C3-8 carbocycle, and 3- to 8-membered heterocycle, wherein the C3-8 carbocycle and 3- to 8-membered heterocycle are each optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, and ═O. In some embodiments, R10 is independently selected at each occurrence from hydrogen and C1-4 alkyl. In some embodiments, R10 is hydrogen.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R11 is independently selected at each occurrence from hydrogen, C1-4 alkyl, C1-4 haloalkyl, C3-8 carbocycle, and 3- to 8-membered heterocycle, wherein the C3-8 carbocycle and 3- to 8-membered heterocycle are each optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, and ═O. In some embodiments, R11 is independently selected at each occurrence from hydrogen, C1-4 alkyl, C3-8 carbocycle, and 3- to 8-membered heterocycle, wherein the C3-8 carbocycle and 3- to 8-membered heterocycle are each optionally substituted with one or more C1-4 alkyl. In some embodiments, R11 is independently selected from: hydrogen, C1-3 alkyl, C1-3 haloalkyl, and C3-4 carbocycle In some embodiments, R11 is independently selected at each occurrence from hydrogen and C1-4 alkyl. In some embodiments, R11 is hydrogen.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R12 is independently selected at each occurrence from hydrogen, C1-4 alkyl, C1-4 haloalkyl, C3-8 carbocycle, and 3- to 8-membered heterocycle, wherein the C3-8 carbocycle and 3- to 8-membered heterocycle are each optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, and ═O. In some embodiments, R12 is independently selected at each occurrence from hydrogen and C1-4 alkyl. In some embodiments, R12 is hydrogen.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R13 is independently selected at each occurrence from hydrogen, C1-4 alkyl, C1-4 haloalkyl, C3-8 carbocycle, and 3- to 8-membered heterocycle, wherein the C3-8 carbocycle and 3- to 8-membered heterocycle are each optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, and ═O. In some embodiments, R13 is independently selected at each occurrence from hydrogen and C1-4 alkyl. In some embodiments, R13 is hydrogen.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R14 is independently selected at each occurrence from hydrogen, C1-4 alkyl, C1-4 haloalkyl, C3-8 carbocycle, and 3- to 8-membered heterocycle, wherein the C3-8 carbocycle and 3- to 8-membered heterocycle are each optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, and ═O. In some embodiments, R14 is independently selected at each occurrence from hydrogen and C1-4 alkyl. In some embodiments, R14 is hydrogen.
In some embodiments, for the compound or salt of Formula (II), (II-A), (II-B), or (II-C), R15 is independently selected at each occurrence from hydrogen, C1-4 alkyl, C1-4 haloalkyl, C3-8 carbocycle, and 3- to 8-membered heterocycle, wherein the C3-8 carbocycle and 3- to 8-membered heterocycle are each optionally substituted with one or more substituents independently selected from halogen, C1-4 alkyl, C1-4 haloalkyl, and ═O. In some embodiments, R15 is independently selected at each occurrence from hydrogen and C1-4 alkyl. In some embodiments, R15 is hydrogen.
In some embodiments, the compounds of Formula (I) and (II) are a compound of Table 1.
While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.
Chemical entities having carbon-carbon double bonds or carbon-nitrogen double bonds may exist in Z- or E- form (or cis- or trans-form). Furthermore, some chemical entities may exist in various tautomeric forms. Unless otherwise specified, compounds or salts of Formula (I), (I-A), (II), (II-A), (II-B), or (II-C), are intended to include all Z-, E- and tautomeric forms as well.
“Isomers” are different compounds that have the same molecular formula. “Stereoisomers” are isomers that differ only in the way the atoms are arranged in space. “Enantiomers” are a pair of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a “racemic” mixture. The term “(±)” is used to designate a racemic mixture where appropriate. “Diastereoisomers” or “diastereomers” are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. The absolute stereochemistry is specified according to the Cahn-Ingold-Prelog R-S system. When a compound is a pure enantiomer, the stereochemistry at each chiral carbon can be specified by either R or S. Resolved compounds whose absolute configuration is unknown can be designated (+) or (−) depending on the direction (dextro- or levorotatory) in which they rotate plane polarized light at the wavelength of the sodium D line. Certain compounds described herein contain one or more asymmetric centers and can thus give rise to enantiomers, diastereomers, and other stereoisomeric forms, the asymmetric centers of which can be defined, in terms of absolute stereochemistry, as (R)- or (S)-. The present chemical entities, pharmaceutical compositions and methods are meant to include all such possible stereoisomers, including racemic mixtures, optically pure forms, mixtures of diastereomers and intermediate mixtures. Optically active (R)- and (S)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. The optical activity of a compound can be analyzed via any suitable method, including but not limited to chiral chromatography and polarimetry, and the degree of predominance of one stereoisomer over the other isomer can be determined.
The compounds or salts for Formula (I), (I-A), (II), (II-A), (II-B), or (II-C) herein may in some cases exist as diastereomers, enantiomers, or other stereoisomeric forms. The compounds presented herein include all diastereomeric, enantiomeric, and epimeric forms as well as the racemates, mixtures of diastereomers, and other mixtures thereof, to the extent they can be made by one of ordinary skill in the art by routine experimentation. Separation of stereoisomers may be performed by chromatography or by forming diastereomers and separating by recrystallization, or chromatography, or any combination thereof (Jean Jacques, Andre Collet, Samuel H. Wilen, “Enantiomers, Racemates and Resolutions”, John Wiley And Sons, Inc., 1981, herein incorporated by reference for this disclosure). Stereoisomers may also be obtained by stereoselective synthesis. Furthermore, a mixture of two enantiomers enriched in one of the two can be purified to provide further optically enriched form of the major enantiomer by recrystallization and/or trituration.
In certain embodiments, compounds or salts for Formula (I), (I-A), (II), (II-A), (II-B), or (II-C), may comprise two or more enantiomers or diatereomers of a compound wherein a single enantiomer or diastereomer accounts for at least about 70% by weight, at least about 80% by weight, at least about 90% by weight, at least about 98% by weight, or at least about 99% by weight or more of the total weight of all stereoisomers. Methods of producing substantially pure enantiomers are well known to those of skill in the art. For example, a single stereoisomer, e.g., an enantiomer, substantially free of its stereoisomer may be obtained by resolution of the racemic mixture using a method such as formation of diastereomers using optically active resolving agents (Stereochemistry of Carbon Compounds, (1962) by E. L. Eliel, McGraw Hill; Lochmuller (1975) J Chromatogr., 113(3): 283-302). Racemic mixtures of chiral compounds can be separated and isolated by any suitable method, including, but not limited to: (1) formation of ionic, diastereomeric salts with chiral compounds and separation by fractional crystallization or other methods, (2) formation of diastereomeric compounds with chiral derivatizing reagents, separation of the diastereomers, and conversion to the pure stereoisomers, and (3) separation of the substantially pure or enriched stereoisomers directly under chiral conditions. Another approach for separation of the enantiomers is to use a Diacel chiral column and elution using an organic mobile phase such as done by Chiral Technologies (www.chiraltech.com) on a fee for service basis.
A “tautomer” refers to a molecule wherein a proton shift from one atom of a molecule to another atom of the same molecule is possible. In certain embodiments, the compounds or salts for Formula (I), (I-A), (II), (II-A), (II-B), or (II-C), exist as tautomers. In circumstances where tautomerization is possible, a chemical equilibrium of the tautomers may exist. The exact ratio of the tautomers depends on several factors, including physical state, temperature, solvent, and pH. Some non-limiting examples of tautomeric equilibrium include:
The compounds disclosed herein, in some embodiments, are used in different enriched isotopic forms, e.g., enriched in the content of 2H, 3H, 11C, 13C and/or 14C. In one particular embodiment, the compound is deuterated in at least one position. Such deuterated forms can be made by the procedure described in U.S. Pat. Nos. 5,846,514 and 6,334,997. As described in U.S. Pat. Nos. 5,846,514 and 6,334,997, deuteration can improve the metabolic stability and or efficacy, thus increasing the duration of action of drugs.
In certain embodiments, the compounds disclosed herein have some or all of the 1H atoms replaced with 2H atoms. The methods of synthesis for deuterium-containing compounds are known in the art and include, by way of non-limiting example only, the following synthetic methods.
Deuterium substituted compounds are synthesized using various methods such as described in: Dean, Dennis C.; Editor. Recent Advances in the Synthesis and Applications of Radiolabeled Compounds for Drug Discovery and Development. [In: Curr., Pharm. Des., 2000; 6(10)]2000, 110 pp; George W.; Varma, Rajender S. The Synthesis of Radiolabeled Compounds via Organometallic Intermediates, Tetrahedron, 1989, 45(21), 6601-21; and Evans, E. Anthony. Synthesis of radiolabeled compounds, J. Radioanal. Chem., 1981, 64(1-2), 9-32.
Deuterated starting materials are readily available and are subjected to the synthetic methods described herein to provide for the synthesis of deuterium-containing compounds. Large numbers of deuterium-containing reagents and building blocks are available commercially from chemical vendors, such as Aldrich Chemical Co.
Unless otherwise stated, compounds described herein are intended to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by 13C- or 14C-enriched carbon are within the scope of the present disclosure.
The compounds of the present disclosure optionally contain unnatural proportions of atomic isotopes at one or more atoms that constitute such compounds. For example, the compounds may be labeled with isotopes, such as for example, deuterium (2H), tritium (H), iodine-125 (125I) or carbon-14 (14C). Isotopic substitution with 2H, 11C, 13C, 14C, 15C, 12N, 13N, 15N, 16N, 16O, 17O, 14F, 15F, 16F, 17F, 18F, 33S34S, 35S, 36S, 35Cl, 37Cl, 79Br, 81Br, and 125I are all contemplated. All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.
Included in the present disclosure are salts, particularly pharmaceutically acceptable salts, of the compounds of Formula (I), (I-A), (II), (II-A), (II-B), or (II-C). The compounds of the present disclosure may possess a sufficiently acidic, a sufficiently basic, or both functional groups, can react with any of a number of inorganic bases, and inorganic and organic acids, to form a salt. Alternatively, compounds that are inherently charged, such as those with a quaternary nitrogen, can form a salt with an appropriate counterion, e.g., a halide such as bromide, chloride, or fluoride, particularly bromide.
The methods and compositions of Formula (I), (I-A), (II), (II-A), (II-B), or (II-C) include the use of amorphous forms as well as crystalline forms (also known as polymorphs). The compounds described herein may be in the form of pharmaceutically acceptable salts. As well, in some embodiments, active metabolites of these compounds having the same type of activity are included in the scope of the present disclosure. In addition, the compounds described herein can exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. The solvated forms of the compounds presented herein are also considered to be disclosed herein.
Compounds of Formula (I), (I-A), (II), (II-A), (II-B), or (II-C), also include crystalline and amorphous forms of those compounds, pharmaceutically acceptable salts, and active metabolites of these compounds having the same type of activity, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), conformational polymorphs, and amorphous forms of the compounds, as well as mixtures thereof.
Included in the present disclosure are salts, particularly pharmaceutically acceptable salts, of compounds represented by Formula (I), (I-A), (II), (II-A), (II-B), or (II-C). The compounds of the present invention that possess a sufficiently acidic, a sufficiently basic, or both functional groups, can react with any of a number of inorganic bases, and inorganic and organic acids, to form a salt. Alternatively, compounds that are inherently charged, such as those with a quaternary nitrogen, can form a salt with an appropriate counterion, e.g., a halide such as bromide, chloride, or fluoride, particularly bromide.
In certain embodiments, compounds or salts of Formula (I), (I-A), (II), (II-A), (II-B), or (II-C), may be prodrugs, e.g., wherein a hydroxyl in the parent compound is presented as an ester or a carbonate, or carboxylic acid present in the parent compound is presented as an ester. The term “prodrug” is intended to encompass compounds which, under physiologic conditions, are converted into pharmaceutical agents of the present disclosure. One method for making a prodrug is to include one or more selected moieties which are hydrolyzed under physiologic conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by an enzymatic activity of the host animal such as specific target cells in the host animal. For example, esters or carbonates (e.g., esters or carbonates of alcohols or carboxylic acids and esters of phosphonic acids) are preferred prodrugs of the present disclosure.
In some aspects, the present disclosure provides a pharmaceutical composition comprising a compound or salt of Formula (I), (I-A), (II), (II-A), (II-B), or (II-C) and at least one pharmaceutically acceptable excipient.
Pharmaceutical compositions can be formulated using one or more physiologically-acceptable carriers comprising excipients and auxiliaries. Formulation can be modified depending upon the route of administration chosen. Pharmaceutical compositions comprising a compound, salt or conjugate can be manufactured, for example, by lyophilizing the compound, salt or conjugate, mixing, dissolving, emulsifying, encapsulating or entrapping the conjugate. The pharmaceutical compositions can also include the compounds, salts or conjugates in a free-base form or pharmaceutically-acceptable salt form.
A compound or salt of Formula (I), (I-A), (II), (II-A), (II-B), or (II-C) may be formulated in any suitable pharmaceutical formulation. A pharmaceutical formulation of the present disclosure typically contains an active ingredient (e.g., compound or salt of any one of Formula (I), (I-A), (II), (II-A), (II-B), or (II-C)), and one or more pharmaceutically acceptable excipients or carriers, including but not limited to: inert solid diluents and fillers, diluents, sterile aqueous solution and various organic solvents, permeation enhancers, antioxidents, solubilizers, and adjuvants.
Pharmaceutical formulations may be provided in any suitable form, which may depend on the route of administration. In some embodiments, the pharmaceutical composition disclosed herein can be formulated in dosage form for administration to a subject. In some embodiments, the pharmaceutical composition is formulated for oral, intravenous, intraarterial, aerosol, parenteral, buccal, topical, transdermal, rectal, intramuscular, subcutaneous, intraosseous, intranasal, intrapulmonary, transmucosal, inhalation, and/or intraperitoneal administration. In some embodiments, the dosage form is formulated for oral administration. For example, the pharmaceutical composition can be formulated in the form of a pill, a tablet, a capsule, an inhaler, a liquid suspension, a liquid emulsion, a gel, or a powder. In some embodiments, the pharmaceutical composition can be formulated as a unit dosage in liquid, gel, semi-liquid, semi-solid, or solid form. Preparations for such pharmaceutical composition are well-known in the art. See, e.g., Anderson, Philip O.; Knoben, James E.; Troutman, William G, eds., Handbook of Clinical Drug Data, Tenth Edition, McGraw-Hill, 2002; Pratt and Taylor, eds., Principles of Drug Action, Third Edition, Churchill Livingston, New York, 1990; Katzung, ed., Basic and Clinical Pharmacology, Ninth Edition, McGraw Hill, 2003; Goodman and Gilman, eds., The Pharmacological Basis of Therapeutics, Tenth Edition, McGraw Hill, 2001; Remingtons Pharmaceutical Sciences, 20th Ed., Lippincott Williams & Wilkins., 2000; Martindale, The Extra Pharmacopoeia, Thirty-Second Edition (The Pharmaceutical Press, London, 1999).
The compounds described herein can be used in the preparation of medicaments for the prevention or treatment of diseases or conditions. In addition, a method for treating any of the diseases or conditions described herein in a subject in need of such treatment, involves administration of pharmaceutical compositions containing at least one compound described herein, or a pharmaceutically acceptable salt, pharmaceutically acceptable prodrug, or pharmaceutically acceptable solvate thereof, in therapeutically effective amounts to said subject.
The compositions containing the compound(s) described herein can be administered for prophylactic and/or therapeutic treatments. In therapeutic applications, the compositions are administered to a patient already suffering from a disease or condition, in an amount sufficient to cure or at least partially arrest the symptoms of the disease or condition. Amounts effective for this use will depend on the severity and course of the disease or condition, previous therapy, the patient's health status, weight, and response to the drugs, and the judgment of the treating physician.
In prophylactic applications, compositions containing the compounds described herein are administered to a patient susceptible to or otherwise at risk of a particular disease, disorder or condition. Such an amount is defined to be a “prophylactically effective amount or dose.” In this use, the precise amounts also depend on the patient's state of health, weight, and the like. When used in a patient, effective amounts for this use will depend on the severity and course of the disease, disorder or condition, previous therapy, the patient's health status and response to the drugs, and the judgment of the treating physician.
In some aspects, the present disclosure provides a method for treatment, comprising administering to a subject in need thereof an effective amount of a compound or salt of Formula (I), (I-A), (II), (II-A), (II-B), or (II-C). In some aspects, the present disclosure provides a method for treating cancer in a patient in need thereof, comprising administering to the subject an effective amount of a compound or salt of Formula (I), (I-A), (II), (II-A), (II-B), or (II-C).
In certain embodiments, the present disclosure can be used as a method of inhibiting an AKT1 protein in a subject in need thereof, comprising administering to the subject a compound or salt of Formula (I), (I-A), (II), (II-A), (II-B), or (II-C) or a pharmaceutical composition of Formula (I), (I-A), (II), (II-A), (II-B), or (II-C). In some embodiments, the AKT protein is a mutant AKT1 protein. In some embodiments, the mutant AKT1 protein comprises an E17K mutant.
The invention now being generally described, it will be more readily understood by reference to the following examples which are included merely for purposes of illustration of certain aspects and embodiments of the present invention, and are not intended to limit the invention in any way.
The following synthetic schemes are provided for purposes of illustration, not limitation. The following examples illustrate the various methods of making compounds described herein. It is understood that one skilled in the art may be able to make these compounds by similar methods or by combining other methods known to one skilled in the art. It is also understood that one skilled in the art would be able to make, in a similar manner as described below by using the appropriate starting materials and modifying the synthetic route as needed. In general, starting materials and reagents can be obtained from commercial vendors or synthesized according to sources known to those skilled in the art or prepared as described herein.
To a solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-chloroimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 1-1) (300 mg, 0.665 mmol, 1 equiv) in 1,4-dioxane (4 mL) and water (1 mL) were added 4-(morpholin-4-yl)phenylboronic acid (207 mg, 0.998 mmol, 1.5 equiv), tetrakis(triphenylphosphine)palladium(0) (77 mg, 0.067 mmol, 0.1 equiv) and sodium carbonate (141 mg, 1.33 mmol, 2 equiv). The resulting mixture was stirred at 90° C. under nitrogen atmosphere for 2 h. The mixture was then concentrated in vacuo and the residue was purified by reverse-phase flash column chromatography on C18 silica gel using a 20-95% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) with a 40 min hold at 70% acetonitrile to afford tert-butyl (4-(2-(2-aminopyridin-3-yl)-5-(4-morpholinophenyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)carbamate (360 mg, 94%) as a yellow solid. MS (ESI) calculated for C33H35N7O3: 577.28 m/z, found 578.55 [M+H]+.
A solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-[4-(morpholin-4-yl)phenyl]imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (416 mg, 0.720 mmol, 1 equiv) in 4N hydrochloric acid in 1,4-dioxane (10 mL) was stirred at room temperature for 1 h. The resulting solution was concentrated in vacuo to afford to afford 3-(3-(4-(aminomethyl)phenyl)-5-(4-morpholinophenyl)-3H-imidazo[4,5-b]pyridin-2-yl)pyridin-2-amine (340 mg, 99% crude) as a yellow solid, which was used in subsequent transformations without further purification. MS (ESI) calculated for C28H27N7O: 477.23 m/z, found 478.25 [M+H]+.
To a solution of 3-(3-(4-(aminomethyl)phenyl)-5-(4-morpholinophenyl)-3H-imidazo[4,5-b]pyridin-2-yl)pyridin-2-amine (180 mg, 0.377 mmol, 1 equiv) in N,N-dimethylformamide (4 mL) were added N,N-diisopropylethylamine (244 mg, 1.89 mmol, 5 equiv), PyBOP (294 mg, 0.566 mmol, 1.5 equiv) and [4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]acetic acid (Intermediate 1-4) (143 mg, 0.415 mmol, 1.1 equiv). The resulting mixture was stirred at room temperature for 1 h. The mixture was purified by reverse-phase flash column chromatography on C18 silica gel using a 20-95% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) with a 40 minute hold at 70% acetonitrile to afford 2-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl)-N-(4-(2-(2-aminopyridin-3-yl)-5-(4-morpholinophenyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)acetamide (153 mg, 50%) as a yellow solid. MS (ESI) calculated for C47H45N7O6: 803.34 m/z, found 804.40 [M+H]+.
A solution of 2-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl)-N-(4-(2-(2-aminopyridin-3-yl)-5-(4-morpholinophenyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)acetamide (100 mg, 0.124 mmol, 1 equiv) in 2,2,2-trifluoroacetic acid (0.6 mL) and methanesulfonic acid (0.2 mL) was stirred at room temperature for 1 h. The resulting mixture was concentrated in vacuo and purified by preparative HPLC on a XSelect CSH C18 OBD Column using a 12-28% gradient of acetonitrile in water (+0.1% formic acid) to afford N-(4-(2-(2-aminopyridin-3-yl)-5-(4-morpholinophenyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)-2-(4-formyl-3-hydroxyphenyl)acetamide (Example 1) (20 mg, 25%) as a yellow solid. MS (ESI) calculated for C37H33N7O4: 639.26 m/z, found 640.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.19 (s, 1H), 8.73-8.79 (m, 1H), 8.12-8.20 (m, 1H), 7.87-8.00 (m, 4H), 7.60-7.63 (m, 1H), 7.41-7.42 (m, 4H), 7.17-7.20 (m, 1H), 6.90-7.03 (m, 4H), 6.41-6.43 (m, 1H), 4.39-4.41 (m, 2H), 3.74-3.77 (m, 4H), 3.56 (s, 2H), 3.16-3.19 (m, 4H).
A solution of tert-butyl N-[(4-aminophenyl)methyl]carbamate (80 g, 360 mmol, 1 equiv), 2,6-dichloro-3-nitropyridine (70 g, 363 mmol, 1 equiv) and N,N-diisopropylethylamine (186 g, 1.44 mol, 4 equiv) in 1,4-dioxane (1.6 L) was stirred at 80° C. overnight. The mixture was allowed to cool to room temperature. The reaction was quenched with water (3 L) and extracted with ethyl acetate (1 L×3). The combined organic layers were washed with brine (1 L), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was re-crystallized from diethyl ether, petroleum ether (1:1, 1 L) to afford tert-butyl N-({4-[(6-chloro-3-nitropyridin-2-yl)amino]phenyl}methyl)carbamate (90 g, 66%) as a red solid. MS (ESI) calculated for C17H19ClN4O4: 378.11 m/z, found 401.05 [M+Na]+.
A solution of tert-butyl N-({4-[(6-chloro-3-nitropyridin-2-yl)amino]phenyl}methyl)carbamate (50 g, 132 mmol, 1 equiv), 2-aminonicotinaldehyde (19.19 g, 158.4 mmol, 1.2 equiv) and sodium dithionite (54.93 g, 386.7 mmol, 2.93 equiv) in dimethyl sulfoxide (750 mL) and methanol (150 mL) was refluxed at 100° C. overnight. The reaction mixture was then poured into water (3 L) and the precipitate was collected by filtration. The solid was recrystallized from methanol/diethyl ether (1:4) to afford tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-chloroimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 1-1) (15.5 g, 25%) as a yellow solid. MS (ESI) calculated for C23H23ClN6O2: 450.16 m/z, found 451.15 [M+H]+.
Synthetic route:
To a solution of 2-(4-bromo-2-((4-methoxybenzyl)oxy)phenyl)-1,3-dioxolane (Intermediate 1-3) (4.15 g, 11.3 mmol, 1 equiv) and tert-butyldimethylsilyl methyl carbonate (8.65 g, 45.4 mmol, 4 equiv) in N,N-dimethylformamide (20 mL) were added bis(tri-tert-butylphosphine)palladium(0) (0.60 g, 1.14 mmol, 0.1 equiv) and lithium fluoride (0.6 g, 23 mmol, 2 equiv). The resulting mixture was stirred at 100° C. under nitrogen atmosphere for 2 h then cooled to 0° C. and quenched with water (100 mL). The resulting mixture was extracted with ethyl acetate (200 mL×3). The combined organic layers were washed with water (100 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-60% gradient of ethyl acetate in petroleum ether to provide methyl 2-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl)acetate (3.0 g, 74%) as a light-yellow oil. MS (ESI) calculated for C20H22O6: 358.14 m/z, found 359.10 [M+H]+.
To a cooled (0° C.) solution of methyl 2-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl)acetate (3.0 g, 8.4 mmol, 1 equiv) in tetrahydrofuran (50 mL) and water (50 mL) was added a solution of lithium hydroxide (0.61 g, 25 mmol, 3 equiv) in water (12.5 mL) and the resulting mixture was stirred at room temperature for 2 h. The mixture was then concentrated in vacuo and suspended in water (5 mL). The pH of the mixture was brought to 6 with 2N hydrochloric acid and the resulting precipitate was collected by filtration and dried to provide 2-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl)acetic acid (Intermediate 1-4) (2.5 g, 87%) as a light-yellow solid. MS (ESI) calculated for C19H20O6: 344.13 m/z, found 345.15 [M+H]+.
To a solution of 5-bromo-2-(1,3-dioxolan-2-yl)phenol (Intermediate 1-2) (5.00 g, 20.4 mmol, 1 equiv) and p-methoxybenzyl chloride (3.83 g, 24.5 mmol, 1.2 equiv) in N,N-dimethylformamide (20 mL) were added potassium iodide (0.340 g, 2.04 mmol, 0.1 equiv) and potassium carbonate (8.46 g, 61.2 mmol, 3 equiv). After stirring overnight at 70° C. under nitrogen atmosphere the resulting mixture was partially concentrated in vacuo and quenched by addition of saturated aqueous ammonium chloride (20 mL) at 0° C. The resulting mixture was extracted with ethyl acetate (30 mL×3). The combined organic layers were washed with water (30 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by silica gel column chromatography using a 0-7% gradient of ethyl acetate in petroleum ether to provide 2-{4-bromo-2-[(4-methoxyphenyl)methoxy]phenyl}-1,3-dioxolane (Intermediate 1-3) (5.8 g, 78%) as a white solid. MS (ESI) calculated for C17H17BrO4: 364.03 m/z, found 365.00 [M+H]+.
To a solution of 4-bromo-2-hydroxybenzaldehyde (10.0 g, 49.7 mmol, 1 equiv) in toluene (100 mL) were added p-toluenesulfonic acid (0.860 g, 4.98 mmol, 0.1 equiv), ethylene glycol (15.44 g, 248.7 mmol, 5 equiv) and triethyl orthoformate (22.12 g, 149.24 mmol, 3 equiv) at room temperature. The resulting solution was stirred at room temperature for 10 min then overnight at 90° C. The solution was cooled to 0° C. and quenched by addition of saturated aqueous ammonium chloride (80 mL). The resulting mixture was extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with water (100 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-50% gradient of ethyl acetate in petroleum ether to provide 5-bromo-2-(1,3-dioxolan-2-yl)phenol (Intermediate 1-2) (6 g, 49%) as a light-yellow oil. MS (ESI) calculated for C9H9BrO3: 244.97 m/z, found 245.95 [M+H]+.
Example 2 was prepared in a manner analogous to Example 1 using 3-acetamidophenylboronic acid in place of 4-(morpholin-4-yl)phenylboronic acid. MS (ESI) calculated for C35H29N7O4: 611.23 m/z, found 612.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): δ 10.17 (s, 1H), 8.30-8.37 (m, 1H), 8.11-8.18 (m, 1H), 8.03-8.07 (m, 1H), 7.88-7.95 (m, 1H), 7.64-7.73 (m, 3H), 7.58-7.63 (m, 1H), 7.47-7.52 (m, 2H), 7.38-7.44 (m, 3H), 6.94-6.98 (m, 1H), 6.88-6.93 (m, 1H), 6.70-6.79 (m, 1H), 4.39 (s, 2H), 3.55 (s, 2H), 2.06 (s, 3H).
Example 3 was prepared in a manner analogous to Example 1 (starting from step 3) using Intermediate 3-1 in place of the amine starting material and Intermediate 3-4 in place of Intermediate 1-4. MS (ESI) calculated for C30H27N7O4: 549.21 m/z, found 550.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 10.32 (s, 1H), 8.27-8.28 (m, 1H), 7.99-8.09 (m, 3H), 7.52-7.55 (m, 1H), 7.39-7.45 (m, 4H), 7.08-7.10 (m, 1H), 6.95-6.97 (m, 1H), 6.71-6.74 (m, 1H), 4.55 (s, 2H), 3.66-3.68 (m, 4H), 3.37-3.42 (m, 4H).
To a solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-chloroimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 1-1) (1.00 g, 2.22 mmol, 1 equiv) in 1,4-dioxane (10 mL) were added morpholine (0.970 g, 11.1 mmol, 5 equiv), tris(dibenzylideneacetone)dipalladium(0) (0.20 g, 0.22 mmol, 0.1 equiv) and RuPhos (0.21 g, 0.44 mmol, 0.2 equiv). The resulting mixture was stirred at 100° C. under nitrogen atmosphere overnight then cooled to room temperature and quenched with water (100 mL). The resulting mixture was extracted with ethyl acetate (3×100 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by reverse-phase flash column chromatography on C18 silica gel using a 20-95% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) with a 10 minute hold at 70% acetonitrile to afford tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-(morpholin-4-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (400 mg, 36%) as a yellow solid. MS (ESI) calculated for C27H31N7O3: 501.25 m/z, found 502.45 [M+H]+.
A solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-(morpholin-4-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (400 mg, 0.797 mmol, 1 equiv) in 4N hydrochloric acid in 1,4-dioxane (20 mL) was stirred at room temperature for 1 h. The resulting solution was concentrated in vacuo to afford 3-{3-[4-(aminomethyl)phenyl]-5-(morpholin-4-yl)imidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 3-1) (300 mg, 94%) as a yellow solid, which was used without purification in subsequent transformations. MS (ESI) calculated for C22H23N7O: 401.20 m/z, found 402.25 [M+H]+.
n-Butyllithium (3.3 mL, 8.2 mmol, 3 equiv) was added dropwise to a cooled (−78° C.) solution of 2-(5-bromo-2-((4-methoxybenzyl)oxy)phenyl)-1,3-dioxolane (Intermediate 3-3) (1.00 g, 2.74 mmol, 1 equiv) in tetrahydrofuran (20 mL) and the resulting solution was stirred at −78° C. for 2 h. Carbon dioxide was then passed through the solution using a needle for 10 min. The reaction was quenched with saturated aqueous ammonium chloride (10 mL) and the mixture was extracted with ethyl acetate (20 mL×3). The combined organic phases were washed with brine (30 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by reverse-phase column chromatography on C18 silica gel using a 5-45% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) to provide 3-(1,3-dioxolan-2-yl)-4-((4-methoxybenzyl)oxy)benzoic acid (Intermediate 3-4) (400 mg, 44%) as a light-yellow oil. MS (ESI) calculated for C18H18O6: 330.11 m/z, found 329.05 [M−H]−.
Intermediate 3-3 was prepared in a manner analogous to Intermediate 1-3 using Intermediate 3-2 in place of Intermediate 1-2. MS (ESI) calculated for C17H17BrO4: 364.03 m/z, found 366.95, 388.95 [M+Na, M+Na+2]+.
Intermediate 3-2 was prepared in a manner analogous to Intermediate 1-2 using 5-bromo-2-hydroxybenzaldehyde in place of 4-bromo-2-hydroxybenzaldehyde. MS (ESI) calculated for C9H9BrO3: 243.97 m/z, found 244.90, 246.90 [M+H, M+H+2]+.
Example 4 was prepared in a manner analogous to Example 1 (starting from step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 4-3 in place of Intermediate 1-4. MS (ESI) calculated for C32H23FN6O3: 558.18 m/z, found 559.15[M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.18 (s, 1H), 8.23-8.25 (m, 1H), 8.05-8.13 (m, 1H), 7.95-8.00 (m, 4H), 7.45-7.52 (m, 6H), 7.43-7.45 (m, 1H), 7.36-7.40 (m, 1H), 6.81-6.84 (m, 1H), 6.42-6.45 (m, 1H), 4.56 (s, 2H). 19F-NMR (400 MHz, DMSO-d6) δ (ppm): −101.75.
A suspension of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-chloroimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 1-1) (40 g, 89 mmol, 1 equiv), phenyl boronic acid (21.63 g, 177.4 mmol, 2 equiv), tetrakis(triphenylphosphine)palladium(0) (10.25 g, 8.871 mmol, 0.1 equiv) and sodium carbonate (18.80 g, 177.4 mmol, 2 equiv) in 1,4-dioxane (400 mL) and water (100 mL) was stirred at 90° C. overnight under nitrogen atmosphere. The reaction mixture was cooled to room temperature and poured into water (2 L). The precipitate was filtered and dried in an oven. The obtained solid was recrystallized from methanol/diethyl ether (1:4) to afford tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate as an orange solid (30 g, 69%). MS (ESI) calculated for C29H28N6O2: 492.23 m/z, found 493.25 [M+H]+.
To a solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (20 g, 41 mmol, 1 equiv) in dichloromethane (200 mL) was added hydrochloric acid (4N in dioxane, 100 mL). The resulting mixture was stirred at room temperature for 6 h and concentrated in vacuo. The resulting residue was recrystallized from diethyl ether to afford 3-{3-[4-(aminomethyl)phenyl]-5-phenylimidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 4-1) as an orange solid (13 g, 82%). MS (ESI) calculated for C24H20N6: 392.17 m/z, found 393.25 [M+H]+.
Intermediate 4-3 was prepared in a manner analogous to Intermediate 3-4 using Intermediate 4-2 in place of Intermediate 3-3. MS (ESI) calculated for C18H17FO6: 348.10 m/z, found 347.10 [M−H]−.
Intermediate 4-2 was prepared in a manner analogous to Intermediate 1-3 (via Intermediate 1-2) starting from 5-bromo-4-fluoro-2-hydroxybenzaldehyde in place of 5-bromo-2-hydroxybenzaldehyde. MS (ESI) calculated for C17H16BrFO4: 382.02 m/z, found 383.05 [M+H]+.
Example 5 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 5-2 in place of Intermediate 1-4. MS (ESI) calculated for C34H25F3N6O3: 622.19 m/z, found 623.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 11.86 (s, 1H), 10.06 (s, 1H), 8.64 (s, 1H), 8.33 (s, 1H), 7.99-8.17 (m, 4H), 7.96-7.98 (m, 1H), 7.60-7.72 (m, 1H), 7.28-7.59 (m, 7H), 7.09-7.20 (m, 1H), 6.60-6.77 (m, 1H), 4.41 (s, 2H), 3.90 (s, 2H). 19F NMR (282 MHz, DMSO-d6) δ (ppm): −52.68.
Intermediate 5-2 was prepared in a manner analogous to Intermediate 1-4 (via Intermediates 1-3 and 1-2) using Intermediate 5-1 in place of 4-bromo-2-hydroxybenzaldehyde. MS (ESI) calculated for C20H19F3O6: 412.11 m/z, found 367.20 [M−H]− (mass of the aldehyde resulting from loss of the ethylene glycol protecting group).
A mixture of 3-bromo-2-(trifluoromethyl)phenol (3.00 g, 12.5 mmol, 1 equiv), magnesium(II) chloride (1.78 g, 18.7 mmol, 1.5 equiv), triethylamine (5.67 g, 56 mmol, 4.5 equiv) and paraformaldehyde (8.97 g, 99.6 mmol, 8 equiv) in tetrahydrofuran (30 mL) was stirred for 2 days at 60° C. The resulting mixture was diluted with water (30 mL) and the resulting mixture was extracted with ethyl acetate (3×30 mL). The combined organic layers were washed with water (2×20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by silica gel column chromatography using a 0-30% gradient of ethyl acetate in petroleum ether to afford 4-bromo-2-hydroxy-3-(trifluoromethyl)benzaldehyde (Intermediate 5-1) (1.1 g, 33%) as a white solid. MS (ESI) calculated for C8H4BrF3O2: 267.93 m/z, found 266.90, 268.90 [M−H, M+2−H]−.
Example 6 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 6-1 in place of the amine starting material. MS (ESI) calculated for C35H33N7O5: 631.25 m/z, found 632.30 [M+H]+. 1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.20 (s, 1H), 8.08-8.12 (m, 1H), 7.95-8.01 (m, 1H), 7.56-7.62 (m, 1H), 7.32-7.40 (m, 5H), 7.09-7.15 (m, 1H), 6.95 (s, 1H), 6.86-6.91 (m, 1H), 6.35-6.41 (m, 1H), 4.34-4.39 (m, 2H), 3.35-3.59 (m, 10H), 2.68 (s, 1H), 2.51 (s, 1H), 2.23-2.35 (m, 1H), 1.41-1.49 (m, 1H), 1.33-1.40 (m, 1H).
To a solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-chloroimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 1-1) (1.00 g, 2.22 mmol, 1 equiv) in 1,4-dioxane (12 mL) and water (3 mL) were added ethyl 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopropane-1-carboxylate (0.64 g, 2.7 mmol, 1.2 equiv), palladium(II) acetate (0.05 g, 0.22 mmol, 0.1 equiv), cesium carbonate (2.17 g, 6.65 mmol, 3 equiv) and bis(adamantan-1-yl)(butyl)phosphanium tetrafluoroborate (0.10 g, 0.22 mmol, 0.1 equiv). The resulting mixture was stirred at 100° C. overnight under nitrogen atmosphere. The mixture was then cooled to room temperature and quenched by addition of water (50 mL). The resulting mixture was extracted with ethyl acetate (3×50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by silica gel column chromatography using a 0-95% gradient of ethyl acetate in petroleum with a 40-minute hold at 50% to provide ethyl 2-[2-(2-aminopyridin-3-yl)-3-(4-{[(tert-butoxycarbonyl)amino]methyl}phenyl)imidazo[4,5-b]pyridin-5-yl]cyclopropane-1-carboxylate (390 mg, 30%) as a yellow solid. MS (ESI) calculated for C29H32N6O4: 528.61 m/z, found 529.35 [M+H]+.
To a solution of ethyl 2-[2-(2-aminopyridin-3-yl)-3-(4-{[(tert-butoxycarbonyl) amino]methyl}phenyl)imidazo[4,5-b]pyridin-5-yl]cyclopropane-1-carboxylate (390 mg, 0.738 mmol, 1 equiv) in tetrahydrofuran (5 mL) and water (5 mL) was added 2N aqueous lithium hydroxide (1.1 mL, 3 equiv). The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated in vacuo and the residue was purified by reverse-phase column chromatography on C18 silica gel using a 0-95% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) to afford 2-[2-(2-aminopyridin-3-yl)-3-(4-{[(tert-butoxycarbonyl)amino]methyl}phenyl)imidazo[4,5-b]pyridin-5-yl]cyclopropane-1-carboxylic acid (289 mg, 70%) as a yellow solid. MS (ESI) calculated for C27H28N6O4: 500.56 m/z, found 501.25 [M+H]+.
To a solution of 2-[2-(2-aminopyridin-3-yl)-3-(4-{[(tert-butoxycarbonyl)amino]methyl}phenyl)imidazo[4,5-b]pyridin-5-yl]cyclopropane-1-carboxylic acid (280 mg, 0.559 mmol, 1 equiv) in N,N-dimethylformamide (5 mL) was added N,N-diisopropylethylamine (217 mg, 1.68 mmol, 3 equiv), HATU (319 mg, 0.839 mmol, 1.5 equiv) and morpholine (73 mg, 0.84 mmol, 1.5 equiv). The resulting mixture was stirred at room temperature for 2 h. The mixture was purified by reverse-phase flash column chromatography on C18 silica gel using a 0-95% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) with a 10 minute hold at 50% acetonitrile to afford tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-[2-(morpholine-4-carbonyl)cyclopropyl]imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (236 mg, 70%) as a yellow solid. MS (ESI) calculated for C31H35N7O4: 569.28 m/z, found 570.35 [M+H]+.
A mixture of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-[2-(morpholine-4-carbonyl)cyclopropyl]imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (230 mg, 0.404 mmol, 1 equiv) in 4N hydrochloric acid in 1,4-dioxane (5 mL) was stirred at room temperature for 2 h. The mixture was concentrated in vacuo to afford 3-{3-[4-(aminomethyl)phenyl]-5-[2-(morpholine-4-carbonyl)cyclopropyl]imidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 6-1) (182 mg, 91%) as a yellow solid. MS (ESI) calculated for C26H27N7O2: 469.22 m/z, found 470.25 [M+H]+.
Example 7 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 7-1 in place of the amine starting material. MS (ESI) calculated for C28H22F2N6O3: 528.17 m/z, found 529.10 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 10.72 (s, 1H), 10.21 (s, 1H), 8.72 (s, 1H), 8.43 (s, 1H), 8.01-8.16 (m, 1H), 7.53-7.58 (m, 4H), 7.36-7.52 (m, 4H), 7.29-7.07 (m, 1H), 7.06-6.84 (m, 2H), 6.69 (s, 1H), 4.37 (s, 2H), 3.54 (s, 2H). 19F NMR (282 MHz, DMSO-d6) δ (ppm): −112.04.
A mixture of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-chloroimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 1-1) (5.00 g, 6.65 mmol, 1 equiv) and phenyl formate (2.12 g, 20.0 mmol, 3 equiv) in toluene was stirred for 12 h at 100° C. under nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography using a 0-20% gradient of ethyl acetate in dichloromethane to afford tert-butyl (4-(5-chloro-2-(2-formamidopyridin-3-yl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)carbamate (2 g, 63%) as a yellow solid. MS (ESI) calculated for C24H23ClN6O3: 478.15 m/z, found 479.10 [M+H]+.
A mixture of tert-butyl N-({4-[5-chloro-2-(2-formamidopyridin-3-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (1.00 g, 2.09 mmol, 1 equiv), palladium(II) acetate (0.05 g, 0.21 mmol, 0.1 equiv) and [4-(diphenylphosphanyl)butyl]diphenylphosphane (0.18 g, 0.42 mmol, 0.2 equiv) in dimethyl sulfoxide (10 mL) was degassed with nitrogen atmosphere. Then tert-butyl isocyanide (0.21 g, 2.5 mmol, 1.2 equiv) was added under nitrogen atmosphere. The resulting suspension was stirred at 120° C. for 12 h under nitrogen atmosphere and cooled to room temperature. Water was added and the resulting precipitate was filtered, washed with water, and dried in an oven. The obtained solid was purified by silica gel column chromatography using a 0-40% gradient of ethyl acetate in dichloromethane to afford tert-butyl N-({4-[2-(2-formamidopyridin-3-yl)-5-formylimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (520 mg, 50%) as a yellow solid. MS (ESI) calculated for C25H24N6O4: 472.19 m/z, found 472.50 [M+H]+.
A solution of tert-butyl N-({4-[2-(2-formamidopyridin-3-yl)-5-formylimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (500 mg, 1.06 mmol, 1 equiv) in dichloromethane (15 mL) was treated with a solution of diethylaminosulfur trifluoride (512 mg, 3.17 mmol, 3 equiv) in dichloromethane (10 mL) at 0° C. under nitrogen atmosphere. The mixture was stirred at room temperature overnight. The resulting mixture was diluted with dichloromethane and washed with brine (2×20 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrate in vacuo to provide tert-butyl N-({4-[5-(difluoromethyl)-2-(2-formamidopyridin-3-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (380 mg, 73%) as a yellow solid, which was used without purification in the next step. MS (ESI) calculated for C25H24F2N6O3: 494.19 m/z, found 495.20 [M+H]+.
A solution of lithium hydroxide (44 mg, 1.8 mmol, 2.5 equiv) in water (4 mL) was added to a solution of tert-butyl N-({4-[5-(difluoromethyl)-2-(2-formamidopyridin-3-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl) carbamate (360 mg, 0.728 mmol, 1 equiv) in tetrahydrofuran (12 mL) and the resulting solution was stirred at room temperature overnight. The solution was then extracted with dichloromethane (3×20 mL). The combined organic layers were washed with brine (50 mL×2), dried with sodium sulfate, filtered, and concentrated in vacuo to afford tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-(difluoromethyl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (334 mg, 98%) as a yellow solid. MS (ESI) calculated for C24H24F2N6O2: 466.19 m/z, found 467.20 [M+H]+.
To a solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-(difluoromethyl) imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (360 mg, 0.772 mmol, 1 equiv) in dichloromethane (5 mL) was added 4N hydrochloric acid in 1,4-dioxane (4 mL) and the resulting suspension was stirred at room temperature for 1 h. The mixture was concentrated in vacuo to provide 3-{3-[4-(aminomethyl)phenyl]-5-(difluoromethyl)imidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 7-1) (310 mg, crude quant.) as a yellow solid, which was used without purification in subsequent transformations. MS (ESI) calculated for C19H16F2N6: 366.14 m/z, found 367.20 [M+H]+.
Example 8 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 8-1 in place of the amine starting material. MS (ESI) calculated for C33H31N7O4: 589.24 m/z, found 590.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.20 (s, 1H), 7.92-7.95 (m, 2H), 7.61-7.63 (m, 1H), 7.32-7.59 (m, 4H), 6.90-7.00 (m, 2H), 6.82-6.89 (m, 1H), 6.77-6.83 (m, 1H), 6.33-6.35 (m, 1H), 4.37-4.41 (m, 4H), 3.75-3.78 (m, 2H), 3.44-3.52 (m, 2H), 2.89-2.92 (m, 2H), 1.73-1.81 (m, 4H).
Intermediate 8-1 was prepared in a manner analogous to Intermediate 3-1 using 8-oxa-3-azabicyclo[3.2.1]octane in place of morpholine. MS (ESI) calculated for C24H25N7O: 427.21 m/z, found 428.21 [M+H]+.
Example 9 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 9-1 in place of the amine starting material. MS (ESI) calculated for C33H31N7O4: 589.24 m/z, found 590.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.20 (s, 1H), 8.70-8.74 (m, 1H), 7.92-7.97 (m, 2H), 7.59-7.62 (m, 1H), 7.29-7.35 (m, 4H), 7.02-7.05 (m, 1H), 6.85-6.95 (m, 3H), 6.33-6.37 (m, 1H), 4.30-4.37 (m, 4H), 3.64-3.67 (m, 2H), 3.52-3.54 (m, 3H), 3.45-3.48 (m, 1H), 1.81-1.93 (m, 4H).
Intermediate 9-1 was prepared in a manner analogous to Intermediate 3-1 using 3-oxa-8-azabicyclo[3.2.1]octane in place of morpholine. MS (ESI) calculated for C24H25N7O: 427.21 m/z, found 428.25 [M+H]+.
Example 10 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 10-1 in place of the amine starting material. MS (ESI) calculated for C30H26N6O4: 534.20 m/z, found 535.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 10.18 (s, 1H), 8.16-8.22 (m, 1H), 7.99-8.05 (m, 1H), 7.56-7.67 (m, 2H), 7.33-7.45 (m, 4H), 6.87-7.00 (m, 3H), 6.71-6.77 (m, 1H), 4.36 (s, 2H), 4.08-4.16 (m, 2H), 0.69-0.76 (m, 2H), 0.63-0.69 (m, 2H).
A suspension of cyclopropanol (1.34 g, 23.0 mmol, 2 equiv) and sodium hydride (0.55 g, 23 mmol, 2 equiv) in tetrahydrofuran (10 mL) was stirred at 0° C. for 0.5 h under nitrogen atmosphere. Then 6-chloro-3-nitropyridin-2-amine (2.00 g, 11.5 mmol, 1 equiv) was added and the mixture was stirred at room temperature for 3 h. The resulting mixture was concentrated in vacuo and purified by silica gel column chromatography using a 0-50% gradient of ethyl acetate in petroleum ether to afford 6-cyclopropoxy-3-nitropyridin-2-amine (1.5 g, 49%) as a yellow solid. MS (ESI) calculated for C8H9N3O3: 195.06 m/z, found 196.00 [M+H]+.
A suspension of 6-cyclopropoxy-3-nitropyridin-2-amine (1.5 g, 7.7 mmol, 1 equiv), tert-butyl N-[(4-bromophenyl)methyl]carbamate (3.30 g, 11.5 mmol, 1.5 equiv), palladium(II) acetate (0.17 g, 0.77 mmol, 0.1 equiv), XantPhos (0.89 g, 1.54 mmol, 0.2 equiv) and cesium carbonate (7.51 g, 23.1 mmol, 3 equiv) in 1,4-dioxane (15 mL) was stirred at 100° C. for 16 h under nitrogen atmosphere. The mixture was concentrated in vacuo and the resulting residue was purified by reverse-phase column chromatography on C18 silica gel using a 5-60% gradient of acetonitrile in water (+0.05% 2,2,2-trifluoroacetic acid) to afford tert-butyl N-({4-[(6-cyclopropoxy-3-nitropyridin-2-yl)amino]phenyl}methyl)carbamate (1.5 g, 38%) as a brown/yellow solid. MS (ESI) calculated for C20H24N4O5: 400.17 m/z, found 401.10 [M+H]+.
A suspension of tert-butyl N-({4-[(6-cyclopropoxy-3-nitropyridin-2-yl)amino]phenyl}methyl)carbamate (1.00 g, 2.50 mmol, 1 equiv), 2-aminopyridine-3-carbaldehyde (0.40 g, 3.2 mmol, 1.3 equiv) and sodium dithionite (0.87 g, 5.0 mmol, 2 equiv) in dimethyl sulfoxide (13 mL) and methanol (2 mL) was stirred at 100° C. for 16 h. The mixture was cooled to room temperature and purified by reverse-phase column chromatography on C18 silica gel using a 5-70% gradient of acetonitrile in water (+0.05% 2,2,2-trifluoroacetic acid) to afford tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-cyclopropoxyimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (360 mg, 18%) as a yellow solid. MS (ESI) calculated for C26H28N6O3: 472.22 m/z, found 473.35 [M+H]+.
A suspension of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-cyclopropoxy imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (360 mg, 0.76 mmol, 1 equiv) in 4N hydrochloric acid in 1,4-dioxane (1 mL) and dichloromethane (5 mL) was stirred at room temperature for 2 h. The mixture was concentrated in vacuo to provide 3-{3-[4-(aminomethyl)phenyl]-5-cyclopropoxyimidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 10-1) (360 mg, 57%) as a yellow solid, which was used without further purification in subsequent transformations. MS (ESI) calculated for C21H20N6O: 372.17 m/z, found 373.05 [M+H]+.
Example 11 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 11-1 in place of Intermediate 1-4 and Intermediate 11-2 in place of the amine starting material. MS (ESI) calculated for C35H28N6O3: 580.22 m/z, found 581.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.10 (s, 1H), 8.23-8.30 (m, 1H), 7.95-8.09 (m, 4H), 7.38-7.55 (m, 9H), 7.18-7.21 (m, 1H), 6.75 (s, 1H), 6.61-6.69 (m, 1H), 6.35-6.40 (m, 1H), 3.70 (s, 2H), 1.19-1.31 (m, 4H).
A mixture of 2-chloro-3-nitro-6-phenylpyridine (1.08 g, 4.60 mmol, 1 equiv), tert-butyl 2-(4-aminophenyl)acetate (1.05 g, 5.06 mmol, 1.1 equiv), tris(dibenzylideneacetone) dipalladium(0) (0.42 g, 0.46 mmol, 0.1 equiv), RuPhos (0.21 g, 0.46 mmol, 0.1 equiv) and sodium carbonate (0.98 g, 9.2 mmol, 2 equiv) in 1,4-dioxane (50 mL) was stirred overnight at 90° C. under nitrogen atmosphere. The mixture was cooled to room temperature and concentrated in vacuo. The resulting residue was purified by silica gel column chromatography using a 0-20% gradient of ethyl acetate in petroleum ether to afford tert-butyl 2-{4-[(3-nitro-6-phenylpyridin-2-yl)amino]phenyl}acetate (1 g, 48%) as brown/yellow solid. MS (ESI) calculated for C23H23N3O4: 405.17 m/z, found 406.10 [M+H]+.
A mixture of tert-butyl 2-{4-[(3-nitro-6-phenylpyridin-2-yl)amino]phenyl}acetate (1.00 g, 2.47 mmol, 1 equiv), 2-aminopyridine-3-carbaldehyde (361 mg, 2.96 mmol, 1.2 equiv) and sodium dithionite (1.07 g, 6.17 mmol, 2.5 equiv) in dimethyl sulfoxide (24 mL) and methanol (2 mL) was stirred overnight at 100° C. The mixture was cooled to room temperature, partially concentrated in vacuo and purified by reverse-flash column chromatography on C18 silica gel using a 10-70% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) to provide tert-butyl 2-{4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]phenyl}acetate (700 mg, 51%) as a reddish brown solid. MS (ESI) calculated for C29H27N5O2: 477.22 m/z, found 478.10 [M+H]+.
A mixture of tert-butyl 2-{4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]phenyl}acetate (400 mg, 0.838 mmol, 1 equiv) in 4N hydrochloric acid in 1,4-dioxane (4.2 mL) was stirred for 1 h at room temperature. The resulting mixture was concentrated in vacuo to afford {4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]phenyl}acetic acid (Intermediate 11-1) (220 mg, 53%) as a yellow solid, which was used without further purification in subsequent transformations. MS (ESI) calculated for C25H19N5O2: 421.15 m/z, found 422.05 [M+H]+.
To a solution of 2-(4-bromo-2-((4-methoxybenzyl)oxy)phenyl)-1,3-dioxolane (Intermediate 1-3) (4.15 g, 11.3 mmol, 1 equiv) and tert-butyldimethylsilyl methyl carbonate (8.65 g, 45.4 mmol, 4 equiv) in N,N-dimethylformamide (20 mL) were added bis(tri-tert-butylphosphine)palladium(0) (0.60 g, 1.14 mmol, 0.1 equiv) and lithium fluoride (0.6 g, 23 mmol, 2 equiv). The resulting mixture was stirred at 100° C. under nitrogen atmosphere for 2 h then cooled to 0° C. and quenched with water (100 mL). The resulting mixture was extracted with ethyl acetate (200 mL×3). The combined organic layers were washed with water (100 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-60% gradient of ethyl acetate in petroleum ether to provide methyl 2-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy) phenyl)acetate (3.0 g, 74%) as a light-yellow oil. MS (ESI) calculated for C20H22O6: 358.14 m/z, found 359.10 [M+H]+.
A mixture of methyl 2-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl)acetate (1.0 g, 2.80 mmol, 1 equiv), ethenyldiphenylsulfanium triflate (0.89 g, 4.2 mmol, 1.5 equiv) and 1,8-diazabicyclo[5.4.0]undec-7-ene (1.27 g, 8.36 mmol, 3.0 equiv) in dimethyl sulfoxide (15 mL) was stirred at room temperature overnight. The reaction mixture was quenched by addition of water (50 mL) and extracted with ethyl acetate (50 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by silica gel column chromatography using a 0-100% gradient of ethyl acetate in petroleum ether to afford methyl 1-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl)cyclopropane-1-carboxylate (800 mg, 75%) as a yellow oil. MS (ESI) calculated for C22H24O6: 384.16 m/z, found 385.10 [M+H]+.
To a solution of methyl 1-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl) cyclopropane-1-carboxylate (800 mg, 2.08 mmol, 1 equiv) in tetrahydrofuran (10 mL) and water (10 mL) was added 2M aqueous lithium hydroxide (3.0 mL, 3 equiv). The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated in vacuo and purified by reverse-phase flash column chromatography on C18 silica gel using a 20-95% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) with a 10-minute hold at 70% acetonitrile to afford 1-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl)cyclopropane-1-carboxylic acid (600 mg, 78%) as a white solid. MS (ESI) calculated for C21H22O6: 370.14 m/z, found 371.10 [M+H]+.
A solution of 1-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl)cyclopropane-1-carboxylic acid (600 mg, 1.62 mmol, 1 equiv), diphenylphosphoryl azide (665 mg, 2.42 mmol, 1.5 equiv), triethylamine (0.49 g, 4.8 mmol, 3.0 equiv) and benzyl alcohol (876 mg, 8.10 mmol, 5.0 equiv) in toluene (20 mL) was stirred for 2 h at 90° C. under nitrogen atmosphere. The mixture was cooled to room temperature and concentrated in vacuo. The resulting residue was purified by silica gel column chromatography using a 0-50% gradient of ethyl acetate in petroleum ether to afford benzyl N-{1-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]cyclopropyl}carbamate (400 mg, 26%) as a yellow oil. MS (ESI) calculated for C28H29NO6: 475.20 m/z, found 476.10 [M+H]+.
To a solution of benzyl N-{1-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl) methoxy]phenyl]cyclopropyl}carbamate (400 mg, 0.841 mmol, 1 equiv) in ethyl acetate (30 mL) was added 20% palladium(II) hydroxide on carbon (40 mg, 0.056 mmol, 0.06 equiv). The reaction was stirred at room temperature for 2 h under an atmosphere of H2. The resulting mixture was filtered, concentrated in vacuo, and purified by silica gel chromatography using a 0-100% gradient of ethyl acetate in petroleum ether to afford 1-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl)cyclopropan-1-amine (Intermediate 11-2) (160 mg, 56%) as a yellow solid. MS (ESI) calculated for C20H23NO4: 341.16 m/z, found 342.10 [M+H]+.
A mixture of 3-{3-[4-(aminomethyl)phenyl]-5-[3-(morpholin-4-yl)phenyl]imidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 12-1) (200 mg, 0.42 mmol, 1 equiv) and N,N-diisopropylethylamine (217 mg, 1.68 mmol, 4 equiv) in N,N-dimethylformamide (2 mL) was stirred for 5 min at room temperature. To this mixture was added 3-fluoro-5-formyl-4-hydroxybenzoic acid (77 mg, 0.42 mmol, 1 equiv) and HATU (159 mg, 0.42 mmol, 1 equiv). The resulting mixture was stirred for 2 h at room temperature. The mixture was purified by reverse-phase flash chromatography on C18 silica gel using a 10-50% gradient of acetonitrile in water to afford N-({4-[2-(2-aminopyridin-3-yl)-5-[3-(morpholin-4-yl)phenyl]imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)-3-fluoro-5-formyl-4-hydroxybenzamide (Example 12) (7.9 mg, 3%) as a yellow solid. MS (ESI) calculated for C36H30FN7O4: 643.23 m/z, found 644.35 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 10.32 (s, 1H), 8.31-8.40 (m, 1H), 8.13-8.20 (m, 1H), 8.01-8.10 (m, 3H), 7.78-7.88 (m, 1H), 7.59-7.65 (m, 1H), 7.46-7.60 (m, 5H), 7.30-7.41 (m, 1H), 6.95-7.09 (m, 1H), 7.78-7.89 (m, 1H), 4.50-4.60 (m, 2H), 3.71-3.80 (m, 4H), 3.12-3.21 (m, 4H).
To a solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-chloroimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 1-1) (200 mg, 0.44 mmol, 1 equiv) and (3-morpholinophenyl)boronic acid (368 mg, 1.78 mmol, 4 equiv) in ethanol (5 mL), toluene (5 mL) and water (1 mL) were added sodium bicarbonate (149 mg, 1.78 mmol, 4 equiv) and tetrakis(triphenylphosphine)palladium(0) (51 mg, 0.044 mmol, 0.1 equiv). After stirring overnight at 90° C. under a nitrogen atmosphere, the mixture was concentrated under reduced pressure and purified by reverse-phase flash column chromatography on C18 silica gel using a 10-50% gradient of acetonitrile in water to afford tert-butyl (4-(2-(2-aminopyridin-3-yl)-5-(3-morpholinophenyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)carbamate (160 mg, 62%) as a yellow solid. MS (ESI) calculated for C33H35N7O3: 577.28 m/z, found 578.10 [M+H]+.
To a solution of tert-butyl (4-(2-(2-aminopyridin-3-yl)-5-(3-morpholinophenyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)carbamate (151 mg, 0.262 mmol, 1 equiv) in 1,4-dioxane (5 mL) was added 4N hydrochloric acid in 1,4-dioxane (1 mL) and the resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated in vacuo to afford 3-(3-(4-(aminomethyl)phenyl)-5-(3-morpholinophenyl)-3H-imidazo[4,5-b]pyridin-2-yl)pyridin-2-amine (Intermediate 12-1) (110 mg, 88%) as a yellow solid, which was used without further purification in subsequent transformations. MS (ESI) calculated for C28H27N7O: 477.23 m/z, found 478.10 [M+H]+.
Example 13 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 12-1 in place of the amine starting material and Intermediate 13-2 in place of Intermediate 1-4. MS (ESI) calculated for C37H32ClN7O4: 673.22 m/z, found 674.30 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.22 (s, 1H), 8.15-8.35 (m, 1H), 7.90-8.10 (m, 2H), 7.60-7.75 (m, 2H), 7.40-7.60 (m, 5H), 7.20-7.40 (m, 2H), 6.90-7.10 (m, 2H), 6.30-6.50 (m, 1H), 4.25-4.55 (m, 2H), 3.74-3.80 (m, 4H), 3.40-3.44 (m, 2H), 3.05-3.25 (m, 4H).
Intermediate 13-2 was prepared in a manner analogous to Intermediate 1-4 using Intermediate 13-1 in place of Intermediate 1-3. MS (ESI) calculated for C19H19ClO6: 378.09 m/z, found 333.09 [M+H-CO2]+.
To a solution of 3-bromo-2-chlorophenol (5 g, 24.102 mmol, 1 equiv) in tetrahydrofuran (50 mL) were added paraformaldehyde (10.86 g, 120.5 mmol, 5 equiv), magnesium(II) chloride (3.44 g, 36.2 mmol, 1.5 equiv) and triethylamine (6.10 g, 60.3 mmol, 2.5 equiv). The resulting suspension was stirred for 2 h at 80° C. The mixture was cooled to room temperature and filtered, rinsing with methanol (50 mL×3). The filtrate was concentrated under reduced pressure and the residue was recrystallized from methanol (10 mL) and ethyl acetate (200 mL) to provide 4-bromo-3-chloro-2-hydroxybenzaldehyde (3 g, 59%) as a light-yellow solid. MS (ESI) calculated for C7H4BrClO2: 233.91 m/z, found 232.95 [M−H]−.
To a solution of 4-bromo-3-chloro-2-hydroxybenzaldehyde (3.00 g, 12.7 mmol, 1 equiv) in toluene (30 mL) was added ethylene glycol (5.56 g, 89.2 mmol, 7 equiv), triethyl orthoformate (5.67 g, 38.2 mmol, 3 equiv) and p-toluenesulfonic acid (0.12 g, 0.64 mmol, 0.05 equiv). The resulting solution was stirred at room temperature for 10 min then at 90° C. overnight. The mixture was concentrated and purified by silica gel column chromatography using a 0-30% gradient of ethyl acetate in petroleum ether to provide 3-bromo-2-chloro-6-(1,3-dioxolan-2-yl)phenol (2 g, 55%) as a yellow oil. MS (ESI) calculated for C9H8BrClO3: 277.93 m/z, found 279.10 [M+H]+.
A mixture of 3-bromo-2-chloro-6-(1,3-dioxolan-2-yl)phenol (2.00 g, 7.16 mmol, 1 equiv), p-methoxybenzyl chloride (1.34 g, 8.59 mmol, 1.2 equiv), potassium iodide (0.89 g, 0.72 mmol, 0.1 equiv) and potassium carbonate (2.97 g, 21.5 mmol, 3 equiv) in N,N-dimethylformamide (20 mL) was stirred at 70° C. overnight under nitrogen atmosphere. The mixture was cooled to room temperature and diluted with water (40 mL). The mixture was extracted with ethyl acetate (50 mL×3). The combined organic phases were washed with brine (100 mL×3), dried with sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-5% gradient of ethyl acetate in petroleum ether to provide 2-(4-bromo-3-chloro-2-((4-methoxybenzyl)oxy)phenyl)-1,3-dioxolane (Intermediate 13-1) (2.4 g, 85%) as a yellow oil. MS (ESI) calculated for C17H16BrClO4: 397.99 m/z, found 398.95 [M+H]+.
Example 14 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 14-1 in place of Intermediate 1-4. MS (ESI) calculated for C34H28N6O3: 568.22 m/z, found 569.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ ppm: 9.94 (s, 1H), 8.66-8.71 (m, 1H), 8.13-8.32 (m, 1H), 7.90-7.09 (m, 4H), 7.37-7.56 (m, 8H), 7.08-7.23 (m, 1H), 6.88-7.00 (m, 1H), 6.30-6.44 (m, 1H), 4.39-4.31 (m, 2H), 3.67 (s, 2H), 2.52 (s, 3H).
Intermediate 14-1 was prepared in a manner analogous to Intermediate 13-2 (via Intermediate 13-1) starting from 3-bromo-2-methylphenol in place of 3-bromo-2-chlorophenol. MS (ESI) calculated for C20H22O6: 358.14 m/z, found 359.15 [M+H]+.
Example 15 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 15-1 in place of Intermediate 1-4. MS (ESI) calculated for C32H23FN6O3: 558.18 m/z, found 559.10 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): δ 10.30 (s, 1H), 8.29-8.34 (m, 1H), 7.99-8.08 (m, 4H), 7.81-7.90 (m, 1H), 7.60-7.65 (m, 1H), 7.39-7.54 (m, 7H), 6.89-6.95 (m, 1H), 6.67-6.74 (m, 1H), 4.58 (s, 2H). 19F NMR (400 MHz, DMSO-d6) δ (ppm): −177.54.
Intermediate 15-1 was prepared in a manner analogous to Intermediate 3-4 (via Intermediates 3-3 and 3-2) starting from 3-bromo-2-fluoro-6-hydroxybenzaldehyde in place of 5-bromo-2-hydroxybenzaldehyde. MS (ESI) calculated for C18H17FO6: 348.10 m/z, found 349.15 [M+H]+.
To a stirred solution of 3-{3-[4-(aminomethyl)phenyl]-5-cyclopropylimidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 16-2) (100 mg, 0.281 mmol, 1 equiv) in anhydrous methanol (1.7 mL) and 1,2-dichloroethane (2 mL) was added 2-{3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)phenyl}acetaldehyde (Intermediate 16-4) (90 mg, 0.28 mmol, 1 equiv) and the mixture was stirred at room temperature for 0.5 h. Then, sodium cyanoborohydride (35 mg, 0.56 mmol, 2 equiv) was added and the mixture was stirred for 2 h. The mixture was concentrated in vacuo and purified by reverse-phase flash column chromatography on C18 silica gel using a 20-70% gradient of acetonitrile in water (+0.05% 2,2,2-trifluoroacetic acid) to provide 3-[3-(4-{[(2-{3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)phenyl}ethyl)amino]methyl}phenyl)-5-cyclopropylimidazo[4,5-b]pyridin-2-yl]pyridin-2-amine (100 mg, 54%) as alight yellow solid. MS (ESI) calculated for C38H46N6O3Si: 662.34 m/z, found 663.35 [M+H]+.
To a stirred solution of 3-[3-(4-{[(2-{3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)phenyl}ethyl)amino]methyl}phenyl)-5-cyclopropylimidazo[4,5-b]pyridin-2-yl]pyridin-2-amine (100 mg, 0.151 mmol, 1 equiv) in anhydrous N,N-dimethylformamide (1.5 mL) were added potassium fluoride (8.8 mg, 0.15 mmol, 1 equiv) and hydrogen bromide (0.2 mL, 30% in water) and the mixture was stirred at room temperature for 0.5 h. The resulting mixture was diluted with water and extracted with dichloromethane (5 mL×3). The combined organic phases were concentrated and the residue was purified by preparative HPLC on a YMC-Actus Triart C18 ExRS column using a 17-42% gradient of acetonitrile in water (+10 mmol/L ammonium bicarbonate) to afford 4-{2-[({4-[2-(2-aminopyridin-3-yl)-5-cyclopropylimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)amino]ethyl}-2-hydroxybenzaldehyde (4-{2-[({4-[2-(2-aminopyridin-3-yl)-5-cyclopropylimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)amino]ethyl}-2-hydroxybenzaldehyde (Example 16) (5.1 mg, 7%) as an off-white solid. MS (ESI) calculated for C30H28N6O2: 504.23 m/z, found 505.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.15 (s, 1H), 8.01-8.04 (m, 1H), 7.94-7.96 (m, 1H), 7.57-7.60 (m, 1H), 7.31-7.45 (m, 2H), 7.23-7.28 (m, 3H), 7.07-7.10 (s, 1H), 6.82-6.85 (s, 2H), 6.30-6.34 (m, 1H), 3.78 (m, 2H), 2.74 (m, 4H), 2.13-2.18 (m, 1H), 0.78-0.95 (m, 4H).
Intermediate 16-2 was prepared in a manner analogous to Intermediate 11-1 using Intermediate 16-1 in place of 2-chloro-3-nitro-6-phenylpyridine and tert-butyl N-[(4-aminophenyl)methyl]carbamate in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C21H20N6: 356.17 m/z, found 357.35 [M+H]+.
To a stirred solution of 6-bromo-2-chloro-3-nitropyridine (4.00 g, 16.8 mmol, 1 equiv) in 1,4-dioxane (40 mL) and water (12 mL) were added cyclopropylboronic acid (2.46 g, 28.6 mmol, 1.7 equiv), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.62 g, 0.84 mmol, 0.05 equiv) and potassium carbonate (4.66 g, 33.7 mmol, 2 equiv). The reaction mixture was stirred overnight at 80° C. under nitrogen atmosphere then cooled to room temperature and quenched with water. The resulting mixture was extracted with ethyl acetate (3×60 mL) and the combined organic layers were washed with brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-100% gradient of ethyl acetate in petroleum ether to afford 2-chloro-6-cyclopropyl-3-nitropyridine (Intermediate 16-1) (2.7 g, 81%) as a red solid. MS (ESI) calculated for C8H7ClN2O2: 198.02 m/z, found 199.11 [M+H]+.
To a solution of 5-bromo-2-(1,3-dioxolan-2-yl)phenoxy(tert-butyl)dimethylsilane (Intermediate 16-3) (1.00 g, 2.78 mmol, 1 equiv) and tributyl(prop-2-en-1-yl)stannane (1.84 g, 5.57 mmol, 2 equiv) in N,N-dimethylformamide (10 mL) was added bis(triphenylphosphine) palladium(II) dichloride (0.20 g, 0.28 mmol, 0.1 equiv). The resulting mixture was stirred for 1 h at 80° C. under nitrogen atmosphere then cooled to room temperature and concentrated in vacuo. The resulting residue was purified by reverse-phase flash column chromatography on C18 silica gel using a 10-40% gradient of acetonitrile in water to afford tert-butyl(2-(1,3-dioxolan-2-yl)-5-(prop-2-en-1-yl)phenoxy)dimethylsilane (0.8 g, 90%) as colorless oil. MS (ESI) calculated for C18H28O3Si: 320.18 m/z, found 321.20 [M+H]+.
To a solution of tert-butyl(2-(1,3-dioxolan-2-yl)-5-(prop-2-en-1-yl)phenoxy) dimethylsilane (1.00 g, 3.12 mmol, 1 equiv) in acetonitrile (3 mL) and water (3 mL) were added osmium tetroxide (2.38 g, 9.36 mmol, 3 equiv) and sodium periodate (2.00 g, 9.36 mmol, 3 equiv). After stirring for 30 min at room temperature, the reaction was quenched by addition of water. The resulting mixture was extracted with ethyl acetate (20 mL×3) and the combined organic layers were washed with water (20 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-25% gradient of ethyl acetate in petroleum ether to provide 2-{3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)phenyl}acetaldehyde (0.2 g, 20%) as a light-yellow oil. MS (ESI) calculated for C17H26O4Si: 322.16 m/z, found 323.15 [M+H]+.
To a solution of 5-bromo-2-(1,3-dioxolan-2-yl)phenol (Intermediate 1-2) (19.00 g, 77.53 mmol, 1 equiv) and imidazole (10.56 g, 155.1 mmol, 2 equiv) in dichloromethane (200 mL) was added tert-butylchlorodimethylsilane (16.36 g, 108.5 mmol, 1.4 equiv). The resulting mixture was stirred overnight at room temperature then cooled to 0° C. and quenched by the addition of water (200 mL). The resulting mixture was extracted with ethyl acetate (300 mL×3) and the combined organic layers were washed with water (300 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-7% gradient of ethyl acetate in petroleum ether to provide 5-bromo-2-(1,3-dioxolan-2-yl)phenoxy(tert-butyl)dimethylsilane (Intermediate 16-3) (22 g, 79%) as a colorless oil. MS (ESI) calculated for C15H23BrO3Si. 358.06 m/z, found 359.05 [M+H]+.
A solution of 2-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl)acetic acid (Intermediate 1-4) (100 mg, 0.333 mmol, 1 equiv) and 2,2,2-trifluoroacetic acid (1 mL) in dichloromethane (2 mL) was stirred for 2 h at room temperature. The resulting mixture was concentrated under reduced pressure to afford (4-formyl-3-hydroxyphenyl) acetic acid (90 mg, crude quant.) as a white solid. MS (ESI) calculated for C9H8O4: 180.04 m/z, found 181.05 [M+H]+.
A solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-chloroimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 1-1) (500 mg, 1.11 mmol, 1 equiv), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (745 mg, 4.44 mmol, 4 equiv), tetrakis(triphenylphosphine)palladium(0) (128 mg, 0.111 mmol, 0.1 equiv), cesium carbonate (373 mg, 4.44 mmol, 4 equiv) in toluene (1 mL), ethanol (5 mL) and water (5 mL) was stirred overnight at 100° C. under nitrogen atmosphere. The mixture was cooled to room temperature and quenched with water (50 mL). The resulting mixture was extracted with ethyl acetate (3×50 mL). The combined organic phases were concentrated under reduced pressure and the residue was purified by reverse-phase flash chromatography on C18 silica gel using a 10-50% gradient of acetonitrile in water (+0.05% 2,2,2-trifluoroacetic acid) to afford tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-(prop-1-en-2-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (400 mg, 79%) as a black solid. MS (ESI) calculated for C26H28N6O2: 456.23 m/z, found 457.23 [M+H]+.
A solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-(prop-1-en-2-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (1.00 g, 2.19 mmol, 1 equiv) and methanesulfonic acid (8 mL) in 1,4-dioxane (4 mL) and water (8 mL) was stirred overnight at 80° C. The mixture was cooled to room temperature and brough to pH 7 with ammonium bicarbonate. The aqueous layer was extracted with ethyl acetate (3×50 mL) and the combined organic phases were concentrated under reduced pressure and purified by reverse-phase flash chromatography on C18 silica gel using a 10-50% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) to provide 2-{3-[4-(aminomethyl)phenyl]-2-(2-aminopyridin-3-yl)imidazo[4,5-b]pyridin-5-yl}propan-2-ol (160 mg, 20%) as a white solid. MS (ESI) calculated for C21H22N6O: 374.19 m/z, found 375.19 [M+H]+.
A solution of 2-(3-(4-(aminomethyl)phenyl)-2-(2-aminopyridin-3-yl)-3H-imidazo[4,5-b]pyridin-5-yl)propan-2-ol (100 mg, 0.267 mmol, 1 equiv), (4-formyl-3-hydroxyphenyl)acetic acid (vide supra) (48 mg, 0.27 mmol, 1 equiv), HATU (102 mg, 0.267 mmol, 1 equiv) and N,N-diisopropylethylamine (69 mg, 0.53 mmol, 2 equiv) in N,N-dimethylformamide (3 mL) was stirred at room temperature for 3 h. The resulting mixture was concentrated under reduced pressure and purified by reverse-phase flash chromatography on C18 silica gel using a 10-50% gradient of acetonitrile in water (+0.05% 2,2,2-trifluoroacetic acid) to provide N-({4-[2-(2-aminopyridin-3-yl)-5-(2-hydroxypropan-2-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)-2-(4-formyl-3-hydroxyphenyl)acetamide (Example 17) (89 mg, 61%) as a yellow solid. MS (ESI) calculated for C30H28N6O4 536.22 m/z, found 537.22 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.17 (s, 1H), 8.18-8.20 (m, 1H), 8.02-8.04 (m, 1H), 7.73-7.75 (m, 1H), 7.59-7.65 (m, 2H), 7.34-7.41 (m, 4H), 6.93-6.95 (m, 1H), 6.89-6.91 (m, 1H), 6.71-6.74 (m, 1H), 4.36 (s, 2H), 3.54-3.59 (m, 2H) 1.39-1.45 (m, 6H).
Example 18 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 18-1 in place of the amine starting material. MS (ESI) calculated for C31H28N6O3: 532.22 m/z, found 533.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.18 (s, 1H), 8.06-8.09 (m, 1H), 7.97-7.99 (m, 1H), 7.60-7.62 (m, 1H), 7.33-7.39 (m, 5H), 7.12-7.16 (m, 1H), 6.89-6.95 (m, 2H), 6.38-6.42 (m, 1H), 4.37 (s, 2H), 3.61 (s, 2H), 1.49 (s, 3H), 1.05-1.06 (m, 2H), 0.77-0.79 (m, 2H).
To a solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-chloroimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 1-1) (500 mg, 1.11 mmol, 1 equiv) in 1,4-dioxane (4 mL) and water (1 mL) were added 4,4,5,5-tetramethyl-2-(1-methylcyclopropyl)-1,3,2-dioxaborolane (114 mg, 1.33 mmol, 1.2 equiv), palladium(II) acetate (25 mg, 0.11 mmol, 0.1 equiv) and cataCXium A (CAS 321921-71-5) (80 mg, 0.22 mmol, 0.2 equiv). The resulting mixture was stirred at 120° C. overnight under nitrogen atmosphere. The mixture was then concentrated in vacuo and purified by reverse-phase flash column chromatography on C18 silica gel using a 20-95% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) with a 10 minute hold at 70% acetonitrile to afford tert-butyl (4-(2-(2-aminopyridin-3-yl)-5-(1-methylcyclopropyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)carbamate (85 mg, 17%) as a yellow solid. MS (ESI) calculated for C27H30N6O2: 470.24 m/z, found 471.25 [M+H]+.
A solution of tert-butyl (4-(2-(2-aminopyridin-3-yl)-5-(1-methylcyclopropyl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)carbamate (85 mg, 0.18 mmol, 1 equiv) in 4N hydrochloric acid in 1,4-dioxane (5 mL) was stirred at room temperature for 1 h. The solvent was removed under reduced pressure to afford 3-(3-(4-(aminomethyl)phenyl)-5-(1-methylcyclopropyl)-3H-imidazo[4,5-b]pyridin-2-yl)pyridin-2-amine (Intermediate 18-1) (60 mg, 71%) as a yellow/green solid, which was used without further purification in subsequent transformations. MS (ESI) calculated for C22H22N6: 370.19 m/z, found 371.15 [M+H]+.
To a stirred solution of 3-{3-[4-(2-aminoethyl)phenyl]-5-[3-(morpholin-4-yl)phenyl]imidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 19-2) (155 mg, 0.315 mmol, 1 equiv) in anhydrous methanol (2 mL) and 1,2-dichloroethane (2 mL) were added 3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)benzaldehyde (Intermediate 19-3) (82 mg, 0.26 mmol, 1 equiv) and sodium cyanoborohydride (33 mg, 0.53 mmol, 2 equiv) and the mixture was stirred at room temperature for 2 h. The mixture was quenched with water and extracted with dichloromethane (3×60 mL). The combined organic layers were washed with brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by reverse-phase flash column chromatography on C18 silica gel using a 20-60% gradient of acetonitrile in water to afford 3-(3-(4-(2-((3-((tert-butyldimethylsilyl)oxy)-4-(1,3-dioxolan-2-yl)benzyl)amino)ethyl)phenyl)-5-(3-morpholinophenyl)-3H-imidazo[4,5-b]pyridin-2-yl)pyridin-2-amine (80 mg, 39%) as a yellow solid. MS (ESI) calculated for C45H53N7O4Si: 783.39 m/z, found 784.41 [M+H]+.
To a stirred solution of 3-(3-(4-(2-((3-((tert-butyldimethylsilyl)oxy)-4-(1,3-dioxolan-2-yl)benzyl)amino)ethyl)phenyl)-5-(3-morpholinophenyl)-3H-imidazo[4,5-b]pyridin-2-yl)pyridin-2-amine (80 mg, 0.10 mmol, 1 equiv) in 2,2,2-trifluoroacetic acid (1 mL) was added methanesulfonic acid (0.3 mL) and the mixture was stirred at room temperature for 0.5 h. The reaction was quenched with water and the pH was brought to 8 with sodium bicarbonate. The resulting mixture was extracted with ethyl acetate (3×60 mL) and the combined organic layers were washed with brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC on a XBridge Prep OBD C18 Column using a 26-45% gradient of acetonitrile in water (+10 mmol/L ammonium bicarbonate) to afford 4-(((4-(2-(2-aminopyridin-3-yl)-5-(3-morpholinophenyl)-3H-imidazo[4,5-b]pyridin-3-yl)phenethyl)amino)methyl)-2-hydroxybenzaldehyde (Example 19) (5.1 mg, 8%) as a yellow solid. MS (ESI) calculated for C37H35N7O3: 625.28 m/z, found 626.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.12 (s, 1H), 8.22-8.25 (m, 1H), 7.95-8.00 (m, 2H), 7.58-7.60 (m, 2H), 7.21-7.47 (m, 7H), 6.89-7.00 (m, 3H), 6.39-6.43 (s, 1H), 3.71 (m, 2H), 3.13-3.15 (m, 4H), 2.53-2.84 (m, 8H).
Intermediate 19-2 was prepared in a manner analogous to Intermediate 12-1 using Intermediate 19-1 in place of Intermediate 1-1. MS (ESI) calculated for C29H29N7O: 491.24 m/z, found 492.35 [M+H]+.
Intermediate 19-1 was prepared in a manner analogous to Intermediate 1-1 using tert-butyl N-[2-(4-aminophenyl)ethyl]carbamate in place of tert-butyl (4-aminobenzyl)carbamate. MS (ESI) calculated for C24H25ClN6O2: 464.17 m/z, found 465.16 [M+H]+.
To a cooled (−78° C.) suspension of 2-{4-bromo-2-[(4-methoxyphenyl)methoxy]phenyl}-1,3-dioxolane (Intermediate 1-4) (1.00 g, 2.74 mmol, 1 equiv) in tetrahydrofuran (20 mL) under nitrogen atmosphere was added dropwise n-butyllithium (2.5M in hexanes, 4.4 mL, 4 equiv). The obtained solution was stirred at −40° C. for 1 h. N,N-dimethylformamide (2 mL) was added dropwise and stirring was continued for 1 h at room temperature. The reaction was quenched by addition of saturated aqueous ammonium chloride (10 mL) and the mixture was extracted with ethyl acetate (10 mL×3). The combined organic phases were washed with brine (20 mL×2), dried with sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a gradient of 0-50% dichloromethane in petroleum ether to afford 4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]benzaldehyde (Intermediate 19-3) (669 mg, 51%) as a yellow oil. MS (ESI) calculated for C18H18O5: 314.12 m/z, found 315.0 [M+H]+.
Example 20 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 12-1 in place of the amine starting material and Intermediate 3-4 in place of Intermediate 1-4. MS (ESI) calculated for C36H31N7O4: 625.24 m/z, found 626.30 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 10.32 (s, 1H), 8.30-8.41 (m, 2H), 8.03-8.16 (m, 3H), 7.75-7.85 (m, 1H), 7.55-7.60 (m, 1H), 7.45-7.65 (m, 5H), 7.28-7.38 (m, 1H), 7.05-7.18 (m, 1H), 6.93-7.05 (m, 1H), 7.78-7.89 (m, 1H), 4.50-4.60 (m, 2H), 3.71-3.80 (m, 4H), 3.12-3.21 (m, 4H).
Example 21 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 12-1 in place of the amine starting material and Intermediate 21-2 in place of Intermediate 1-4. MS (ESI) calculated for C37H32FN7O4: 657.25 m/z, found 658.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.18 (s, 1H), 8.30-8.32 (m, 1H), 8.03-8.05 (m, 2H), 7.81-7.82 (m, 1H), 7.42-7.58 (m, 7H), 7.31-7.35 (m, 1H), 6.95-7.02 (m, 2H), 6.85-6.92 (m, 1H), 4.39 (s, 2H), 3.76-3.82 (m, 6H), 3.15-3.16 (m, 4H). 19F NMR (400 MHz, DMSO-d6) δ (ppm): 139.31.
Intermediate 21-2 was prepared in a manner analogous to Intermediate 1-4 using Intermediate 21-1 in place of Intermediate 1-3. MS (ESI) calculated for C19H19FO6: 362.12 m/z, found 363.12 [M+H]+.
Intermediate 21-1 was prepared in a manner analogous to Intermediate 1-3 (via Intermediate 1-2) starting from 3-bromo-2-fluorophenol in place of 4-bromo-2-hydroxybenzaldehyde. MS (ESI) calculated for C17H16BrFO4: 382.02 m/z, found 381.00 [M−H]−
Example 22 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 16-2 in place of the amine starting material and Intermediate 13-2 in place of Intermediate 1-4. MS (ESI) calculated for C30H25ClN6O3: 552.02 m/z, found 553.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.95 (s, 1H), 8.01-8.11 (m, 2H), 7.59-7.75 (m, 2H), 7.35-7.39 (m, 4H), 7.29-7.34 (m, 1H), 7.05-7.15 (m, 1H), 6.71-6.79 (m, 1H), 4.45 (s, 2H), 3.80 (s, 2H), 2.15-2.26 (m, 1H), 0.92-1.01 (m, 2H), 0.79-0.88 (m, 2H).
Example 23 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 23-1 in place of the amine starting material. MS (ESI) calculated for C30H28N6O3: 520.22 m/z, found 521.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.22 (s, 1H), 8.10-8.20 (m, 1H), 7.85-8.02 (m, 1H), 7.50-7.70 (m, 1H), 7.40 (s, 4H), 7.20-7.35 (m, 1H), 7.10-7.20 (m, 1H), 6.92-7.00 (m, 1H), 6.80-6.92 (m, 1H), 6.25-6.45 (m, 1H), 4.30-4.50 (m, 2H), 3.50-3.70 (m, 2H), 3.00-3.15 (m, 1H), 1.15-1.25 (m, 6H).
A solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-chloroimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 1-1) (500 mg, 1.11 mmol, 1 equiv), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (745 mg, 4.44 mmol, 4 equiv), tetrakis(triphenylphosphine)palladium(0) (128 mg, 0.111 mmol, 0.1 equiv), cesium carbonate (373 mg, 4.44 mmol, 4 equiv) in toluene (1 mL), ethanol (5 mL) and water (5 mL) was stirred overnight at 100° C. under nitrogen atmosphere. The mixture was cooled to room temperature and quenched with water (50 mL). The resulting mixture was extracted with ethyl acetate (3×50 mL). The combined organic phases were concentrated under reduced pressure and the residue was purified by reverse-phase flash chromatography on C18 silica gel using a 10-50% gradient of acetonitrile in water (+0.05% 2,2,2-trifluoroacetic acid) to afford tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-(prop-1-en-2-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (400 mg, 79%) as a black solid. MS (ESI) calculated for C26H28N6O2: 456.23 m/z, found 457.23 [M+H]+.
To a solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-(prop-1-en-2-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (214 mg, 0.469 mmol, 1 equiv) in methanol (10 mL) was added Pd(OH)2/C (10%, 22 mg, 0.05 equiv). The mixture was stirred at room temperature under hydrogen atmosphere for 2 h. The mixture was then filtered and concentrated in vacuo to afford tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-isopropylimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (120 mg, 55%) as a yellow solid, which was used without further purification in the next step. MS (ESI) calculated for C26H30N6O2: 458.24 m/z, found 459.15 [M+H]+.
tert-Butyl N-({4-[2-(2-aminopyridin-3-yl)-5-isopropylimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (120 mg, 0.262 mmol, 1 equiv) was dissolved in 4N hydrochloric acid in 1,4-dioxane (5 mL) and the mixture was stirred at room temperature for 1 h. The mixture was concentrated in vacuo to afford 3-{3-[4-(aminomethyl)phenyl]-5-isopropylimidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 23-1) (95 mg, 95%) as a yellow solid, which was used in subsequent transformations without further purification. MS (ESI) calculated for C21H22N6: 358.19 m/z, found 359.35 [M+H]+.
Example 24 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 24-2 in place of the amine starting material. MS (ESI) calculated for C28H24N6O3: 492.19 m/z, found 493.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.17 (s, 1H), 8.05-8.07 (m, 1H), 7.95-7.96 (m, 1H), 7.55-7.61 (m, 1H), 7.35-7.38 (m, 4H), 7.32-7.34 (m, 1H), 7.24-7.26 (m, 1H), 6.94-7.174 (m, 2H), 6.36-6.39 (m, 1H), 4.35-4.36 (m, 2H), 3.51-3.60 (m, 2H), 2.52-2.54 (m, 3H).
A mixture of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-methylimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 24-1) (172 mg, 0.40 mmol, 1 equiv) in 4N hydrochloric acid in 1,4-dioxane (10 mL) was stirred at room temperature for 2 h. The mixture was concentrated in vacuo to afford 3-{3-[4-(aminomethyl)phenyl]-5-methylimidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 24-2) (150 mg, crude quant.) as a yellow solid, which was used without further purification in subsequent transformations. MS (ESI) calculated for C19H18N6: 330.21 m/z, found 331.21 [M+H]+.
Intermediate 24-1 was prepared in a manner analogous to Intermediate 1-1 using 2-chloro-6-methyl-3-nitropyridine in place of 2,6-dichloro-3-nitropyridine. MS (ESI) calculated for C24H26N6O2: 430.21 m/z, found 431.20 [M+H]+.
Example 25 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 25-1 in place of the amine starting material. MS (ESI) calculated for C30H27N7O3: 533.22 m/z, found 534.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ ppm: 10.18 (s, 1H), 7.88-7.91 (m, 2H), 7.55-7.71 (m, 1H), 7.26-7.40 (m, 4H), 7.00-7.06 (m, 1H), 6.90-6.94 (m, 1H), 6.88-6.92 (m, 1H), 6.68-6.75 (m, 1H), 6.29-6.33 (m, 1H), 4.34 (s, 2H), 3.53-3.55 (m, 2H), 2.44-2.47 (m, 1H), 0.66-0.67 (m, 2H), 0.40-0.41 (m, 2H).
To a solution of tert-butyl (4-(2-(2-aminopyridin-3-yl)-5-chloro-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)carbamate (Intermediate 1-1) (252 mg, 0.541 mmol, 1 equiv) in 1,4-dioxane (5 mL) were added aminocyclopropane (154 mg, 2.71 mmol, 5 equiv), tris(dibenzylideneacetone)dipalladium(0) (50 mg, 0.054 mmol, 0.1 equiv), RuPhos (50 mg, 0.11 mmol, 0.2 equiv) and cesium carbonate (441 mg, 1.35 mmol, 2.5 equiv). The resulting mixture was stirred for 4 h at 100° C. under a nitrogen atmosphere, then cooled to room temperature and concentrated in vacuo. The residue was purified by reverse-phase flash column chromatography on C18 silica gel using a 0-40% gradient of acetonitrile in water to afford tert-butyl (4-(2-(2-aminopyridin-3-yl)-5-(cyclopropylamino)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)carbamate (40 mg, 15%) as a yellow solid. MS (ESI) calculated for C26H29N7O2: 471.24 m/z, found 472.40 [M+H]+.
A solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-(cyclopropylamino)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (47 mg, 0.10 mmol, 1 equiv) in 4N hydrochloric acid in 1,4-dioxane was stirred at room temperature for 1 h. The resulting mixture was concentrated to afford 3-{3-[4-(aminomethyl)phenyl]-5-(cyclopropylamino)imidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 25-1) (40 mg, 81%) as a light-yellow solid, which was used without further purification in subsequent transformations. MS (ESI) calculated for C21H21N7: 371.19 m/z, found 372.20 [M+H]+.
Example 26 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 16-2 in place of the amine starting material and Intermediate 65-1 in place of Intermediate 1-4. MS (ESI) calculated for C30H25FN6O3: 536.20 m/z, found 537.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.26 (s, 1H), 9.20-9.23 (m, 1H), 8.07-8.09 (m, 1H), 8.00-8.02 (m, 1H), 7.70-7.72 (m, 1H), 7.57-7.59 (m, 1H), 7.30-7.39 (m, 5H), 7.13-7.20 (m, 1H), 7.05-7.07 (m, 1H), 6.67-6.70 (m, 1H), 5.97-6.09 (m, 1H), 4.41-4.50 (m, 2H), 2.17-2.21 (m, 1H), 0.97-0.98 (m, 2H), 0.95-0.96 (m, 2H). 19F NMR (400 MHz, DMSO-d6) δ (ppm): −181.39.
Example 27 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 16-2 in place of the amine starting material and Intermediate 3-4 in place of Intermediate 1-4. MS (ESI) calculated for C29H24N6O3: 504.19 m/z, found 505.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.32 (s, 1H), 8.28-8.29 (m, 1H), 8.03-8.11 (m, 3H), 7.66-7.69 (m, 1H), 7.39-7.46 (m, 4H), 7.33-7.38 (m, 1H), 7.08-7.11 (m, 1H), 6.75-6.79 (m, 1H), 4.55 (s, 2H), 2.17-2.22 (m, 1H), 0.83-0.96 (m, 4H).
Example 28 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 28-1 in place of the amine starting material and Intermediate 28-2 in place of Intermediate 1-4. MS (ESI) calculated for C32H24N6O3: 540.19 m/z, found 541.30 [M+H]+. 1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.22 (s, 1H), 8.29-8.38 (m, 1H), 8.01-8.09 (m, 4H), 7.75-7.81 (m, 2H), 7.59-7.68 (m, 2H), 7.35-7.52 (m, 5H), 6.95-7.01 (m, 2H), 6.69-6.78 (m, 1H), 3.70 (s, 2H).
Intermediate 28-1 was prepared in a manner analogous to Intermediate 11-1 using tert-butyl N-(4-aminophenyl)carbamate in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C23H18N6: 378.16 m/z, found 379.15 [M+H]+.
Intermediate 28-2 was prepared in a manner analogous to Intermediate 1-4 using Intermediate 3-3 in place of Intermediate 1-3. MS (ESI) calculated for C19H20O6: 344.13 m/z, found 345.15 [M+H]+.
To a solution of 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]benzoic acid (Intermediate 29-2) (85 mg, 0.21 mmol, 1 equiv), 1-(1-benzofuran-5-yl)methanamine (Intermediate 29-3) (37 mg, 0.25 mmol, 1.2 equiv) and N,N-diisopropylethylamine (162 mg, 1.25 mmol, 6 equiv) in N,N-dimethylformamide (2 mL) was added HATU (119 mg, 0.314 mmol, 1.5 equiv). The resulting mixture was stirred at room temperature for 2 h. The reaction was quenched by addition of water (10 mL) and the mixture was extracted with ethyl acetate (10 mL×3). The combined organic phases were washed with brine (30 mL×3), dried with sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-10% gradient of dichloromethane in methanol to afford 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]-N-(1-benzofuran-6-ylmethyl)benzamide (42 mg, 33%) as a yellow solid. MS (ESI) calculated for C33H24N6O2: 536.20 m/z, found 537.05 [M+H]+.
To a solution of 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]-N-(1-benzofuran-6-ylmethyl)benzamide (32 mg, 0.060 mmol, 1 equiv) in acetonitrile (1.2 mL), water (0.4 mL) and 2,2,2-trifluoroacetic acid (14 mg, 0.12 mmol, 2 equiv) was added citric acid (11 mg, 0.060 mmol, 1 equiv), sodium periodate (51 mg, 0.24 mmol, 4 equiv) and ruthenium (III) chloride (0.6 mg, 3 umol, 0.05 equiv). The resulting mixture was stirred at room temperature for 2.5 h. The mixture was concentrated in vacuo and purified by preparative HPLC on a XBridge Prep OBD C18 Column using a 30-60% gradient of acetonitrile in water to afford 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]-N-[(3-formyl-4-hydroxyphenyl)methyl]benzamide (Example 29) (0.5 mg, 1.5%) as a yellow solid. MS (ESI) calculated for C32H24N6O3: 540.19 m/z, found 541.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 6 10.26 (s, 1H), 8.30-8.36 (m, 1H), 7.99-8.13 (m, 7H), 7.59-7.70 (m, 4H), 7.38-7.57 (m, 5H), 6.97-7.02 (m, 1H), 6.69-6.74 (m, 1H), 4.44 (s, 2H).
A solution of 3-(3-(4-bromophenyl)-5-phenyl-3H-imidazo[4,5-b]pyridin-2-yl)pyridin-2-amine (Intermediate 29-1) (500 mg, 1.37 mmol, 1 equiv), phenyl formate (500 mg, 4.10 mmol, 3 equiv), palladium(II) acetate (15 mg, 0.068 mmol, 0.05 equiv), XantPhos (40 mg, 0.068 mmol, 0.05 equiv) and N,N-diisopropylethylamine (353 mg, 2.73 mmol, 2 equiv) in toluene (5 mL) was stirred under nitrogen atmosphere at 90° C. overnight. The resulting mixture was partitioned between ethyl acetate (10 mL) and water (10 mL). The layers were separated, and the aqueous layer was extracted with ethyl acetate (10 mL×2). The combined organic layers were washed with brine (30 mL×3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford phenyl 4-[2-(2-formamidopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]benzoate (570 mg, 20%) as a brown oil, which was used in the next step without further purification. MS (ESI) calculated for C31H21N5O3: 511.16 m/z, found 512.05 [M+H]+.
To a solution of phenyl 4-[2-(2-formamidopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]benzoate (570 mg, 1.11 mmol, 1 equiv) in methanol (1 mL), water (1 mL) and tetrahydrofuran (5 mL) was added lithium hydroxide (80 mg, 3.3 mmol, 3 equiv) and the resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated in vacuo and water (2 mL) was added. The pH of the mixture was brought to 3˜4 with 2N hydrochloric acid and extracted with chloroform/isopropanol (3:1, 5 mL×3). The combined organic phases were concentrated in vacuo and the residue was recrystallized from diethyl ether (5 mL) to afford 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]benzoic acid (Intermediate 29-2) (85 mg, 12%) as a yellow solid. MS (ESI) calculated for C24H17N5O2: 407.14 m/z, found 408.15 [M+H]+.
Intermediate 29-1 was prepared in a manner analogous to Intermediate 11-1 (Steps 1 and 2 only) using 4-bromoaniline in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C23H16BrN5: 441.06 m/z, found 442.00 [M+H]+.
A solution of 5-bromo-1-benzofuran (1.00 g, 5.08 mmol, 1 equiv), tert-butyl ((trifluoro-λ4-boraneyl)methyl)carbamate, potassium salt (1.80 g, 7.61 mmol, 1.5 equiv), tris(dibenzylideneacetone)dipalladium(0) (465 mg, 0.508 mmol, 0.1 equiv), SPhos (208 mg, 0.508 mmol, 0.1 equiv) and cesium carbonate (3.30 g, 10.2 mmol, 2 equiv) in 1,4-dioxane (40 mL) and water (10 mL) was stirred at 80° C. for 16 h. The resulting mixture was cooled to room temperature and quenched with water (10 mL). The mixture was extracted with ethyl acetate (500 mL×2) and the combined organic phases were washed with brine (30 mL×3), dried with sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-10% gradient of ethyl acetate in petroleum ether to afford tert-butyl N-(1-benzofuran-5-ylmethyl)carbamate (800 mg, 55%) as a yellow oil. MS (ESI) calculated for C14H17NO3: 247.12 m/z, found 192.00 [M−C(CH3)3+H]+.
To a solution of tert-butyl N-(1-benzofuran-5-ylmethyl)carbamate (800 mg, 3.24 mmol, 1 equiv) in dichloromethane (5 mL) was added 4N hydrochloric acid in 1,4-dioxane (5 mL) and the resulting mixture was stirred at room temperature for 1 h. The mixture was concentrated in vacuo to provide benzofuran-5-ylmethanamine (Intermediate 29-3) (600 mg, 67%) as a white solid. MS (ESI) calculated for C9H9NO: 147.07 m/z, found 148.10 [M+H]+.
Example 30 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 30-2 in place of the amine starting material and Intermediate 21-2 in place of Intermediate 1-4. MS (ESI) calculated for C34H27FN6O4: 602.21 m/z, found 603.30 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 10.22 (s, 1H), 8.64-8.74 (m, 1H), 8.23-8.34 (m, 1H), 7.93-8.10 (m, 4H), 7.36-7.55 (m, 6H), 7.13-7.21 (m, 1H), 6.96-7.02 (s, 1H), 6.85-6.95 (m, 1H), 6.32-6.44 (m, 2H), 4.94-5.08 (m, 1H), 3.56-3.77 (m, 4H). 19F NMR (282 MHz, DMSO-d6) δ (ppm): −130.09.
Intermediate 30-2 was prepared in a manner analogous to Intermediate 11-1 using Intermediate 30-1 in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C25H22N6O: 422.19 m/z, found 423.20 [M+H]+.
A solution of methyl 2-(4-nitrophenyl)acetate (5.00 g, 25.6 mmol, 1 equiv) in diethyl ether (50 mL) was treated with a solution of isopentyl nitrite (6.60 g, 56.4 mmol, 2.2 equiv) in diethyl ether (35 mL) at room temperature under nitrogen atmosphere followed by dropwise addition of a suspension of sodium methoxide (2.21 g, 41.0 mmol, 1.6 equiv) in methanol (35 mL). The mixture was stirred for 12 h at room temperature. The residue was concentrated and added to 2 N hydrochloric acid followed by stirring at room temperature for 30 min. The solid was filtered and washed with methanol to provide methyl (2E)-2-(N-hydroxyimino)-2-(4-nitrophenyl)acetate (3.1 g, 54%) a white solid. MS (ESI) calculated for C9H8N2O5: 224.17 m/z, found 223.00 [M−H]−.
To a cooled (0° C.) solution of methyl (2E)-2-(N-hydroxyimino)-2-(4-nitrophenyl)acetate (2.9 g, 13 mmol, 1 equiv) in tetrahydrofuran (50 mL) under nitrogen atmosphere was added sodium borohydride (1.47 g, 38.9 mmol, 3 equiv) followed by a solution of iodine (5.00 g, 19.7 mmol, 1.5 equiv) in tetrahydrofuran (33 mL) dropwise. The reaction mixture was stirred at 50° C. for 2 h, then cooled to 0° C. and quenched with water. Stirring was continued for 30 min and the solution was alkalized to pH 9 with 1N aqueous sodium hydroxide. The resulting mixture was used directly in the next step. MS (ESI) calculated for C8H10N2O3: 182.07 m/z, found 183.10 [M+H]+.
To the reaction mixture from the previous step was added di-tert-butyl dicarbonate (12.5 mL, 58.4 mmol, 4.5 equiv) and the mixture was stirred at room temperature for 2 h. The resulting mixture was extracted with ethyl acetate (3×50 mL) and the combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to provide tert-butyl N-[2-hydroxy-1-(4-nitrophenyl)ethyl]carbamate (2.5 g, 81%) as a yellow oil, which was used without purification in the next step. MS (ESI) calculated for C13H18N2O5: 282.12 m/z, found 183.10 [M−Boc+H]+.
A mixture of tert-butyl N-[2-hydroxy-1-(4-nitrophenyl)ethyl]carbamate (2.70 g, 9.56 mmol, 1 equiv) and 10% palladium on carbon (2.70 g, 25.3 mmol, 2.65 equiv) was stirred for 1 day at room temperature under hydrogen atmosphere. The mixture was filtered and concentrated in vacuo to provide tert-butyl N-[1-(4-aminophenyl)-2-hydroxyethyl]carbamate (Intermediate 30-1) (2 g, 83%) as a white oil, which was used in subsequent transformations directly without further purification. MS (ESI) calculated for C13H20N2O3: 252.15 m/z, found 253.05 [M+H]+.
Example 31 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 31-2 in place of the amine starting material and Intermediate 21-2 in place of Intermediate 1-4. MS (ESI) calculated for C35H27FN6O3: 598.21 m/z, found 599.25 [M+H]+. 1H-NMR (300 MHz, DMSO-d6) δ (ppm): 10.19 (s, 1H), 8.25-8.28 (m, 1H), 7.96-8.03 (m, 4H), 7.37-7.50 (m, 2H), 7.25-7.28 (m, 4H), 7.18-7.22 (m, 3H), 6.75-6.81 (m, 1H), 6.40-6.45 (m, 1H), 3.61 (s, 2H), 1.30-1.33 (m, 2H), 1.23-1.26 (m, 2H).
Intermediate 31-2 was prepared in a manner analogous to Intermediate 11-1 using Intermediate 31-1 in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C26H22N6: 418.19 m/z, found 419.25 [M+H]+.
To a solution of 1-(4-bromophenyl)cyclopropan-1-amine (5.00 g, 23.6 mmol, 1 equiv) in dichloromethane (50 mL) were added triethylamine (7.16 g, 70.7 mmol, 3 equiv) and di-tert-butyl dicarbonate (10.29 g, 47.15 mmol, 2 equiv). The resulting mixture was stirred at room temperature for 2 h. The reaction was quenched with water (100 mL) and extracted with ethyl acetate (3×100 mL). The organic layers were combined, dried over sodium sulfate, filtered, and concentrated in vacuo. The obtained residue was purified by silica gel column chromatography using a 0-30% gradient of ethyl acetate in petroleum ether to afford tert-butyl N-[1-(4-bromophenyl)cyclopropyl]carbamate (5 g, 68%) as a yellow solid. MS (ESI) calculated for C14H18BrNO2: 311.05 m/z, found 310.00 [M−H]−.
To a solution of tert-butyl N-[1-(4-bromophenyl)cyclopropyl]carbamate (1.8 g, 5.8 mmol, 1 equiv) in 1,4-dioxane (10 mL) were added O-benzyl carbamate (1.05 g, 6.92 mmol, 1.2 equiv), XantPhos (667 mg, 1.15 mmol, 0.2 equiv), tris(dibenzylideneacetone)dipalladium(0) (528 mg, 0.577 mmol, 0.1 equiv) and cesium carbonate (5.64 g, 17.3 mmol, 3 equiv). The resulting mixture was stirred for 3 h at 100° C. under nitrogen atmosphere. The mixture was then cooled to room temperature and quenched with water (100 mL). The resulting mixture was extracted with ethyl acetate (3×100 mL). The organic layers were combined, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue obtained was purified by silica gel column chromatography using a 0-30% gradient of ethyl acetate in petroleum ether to afford benzyl N-(4-{1-[(tert-butoxycarbonyl)amino]cyclopropyl}phenyl)carbamate (1.1 g, 50%) as a yellow solid. MS (ESI) calculated for C22H26N2O4: 382.19 m/z, found 381.05 [M−H]−.
To a solution of benzyl N-(4-{1-[(tert-butoxycarbonyl)amino]cyclopropyl}phenyl) carbamate (1.1 g, 2.9 mmol, 1 equiv) in methanol (3 mL) was added palladium on carbon (10%, 110 mg, 0.03 equiv) under nitrogen atmosphere. The mixture stirred at room temperature under hydrogen atmosphere for 1 h then filtered and concentrated in vacuo. The residue obtained was purified by silica gel column chromatography using a 0-100% gradient of ethyl acetate in petroleum ether to afford tert-butyl N-[1-(4-aminophenyl)cyclopropyl]carbamate (Intermediate 31-1) (500 mg, 70%) as a yellow solid. MS (ESI) calculated for C14H20N2O2: 248.15 m/z, found 247.00 [M−H]−.
Example 32 was prepared in a manner analogous to Example 1 using 1-methylpyrazol-4-ylboronic acid in place of 4-(morpholin-4-yl)phenylboronic acid. MS (ESI) calculated for C31H26N8O3: 558.21 m/z, found 559.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.17 (s, 1H), 8.12-8.14 (m, 2H), 7.97-7.98 (m, 2H), 7.65-7.67 (m, 2H), 7.36-7.67 (m, 4H), 7.15-7.17 (m, 1H), 6.89-6.94 (m, 2H), 6.37-6.40 (m, 1H), 4.36-4.37 (m, 2H), 3.81-3.88 (m, 3H), 3.48-3.52 (m, 2H).
Example 33 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 33-1 in place of the amine starting material. MS (ESI) calculated for C28H24N6O4: 508.19 m/z, found 509.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 10.14 (s, 1H), 8.08-8.16 (m, 1H), 8.03-7.98 (m, 1H), 7.30-7.54 (m, 2H), 7.30-7.45 (m, 4H), 6.81-7.02 (m, 3H), 6.62-6.80 (m, 1H), 4.30-4.41 (m, 2H), 3.65-3.95 (m, 3H), 3.53-3.67 (m, 2H).
Intermediate 33-2 was prepared in a manner analogous to Intermediate 11-1 using 2-chloro-6-methoxy-3-nitropyridine in place of 2-chloro-3-nitro-6-phenylpyridine and tert-butyl N-[(4-aminophenyl)methyl]carbamate in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C19H18N6O: 346.15 m/z, found 347.39 [M+H]+.
Example 34 was prepared in a manner analogous to Example 1 using 4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl]morpholine in place of 4-(morpholin-4-yl)phenylboronic acid. MS (ESI) calculated for C36H32N8O4: 640.25 m/z, found 641.30 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.22 (s, 1H), 8.40-8.50 (m, 1H), 8.25-8.40 (m, 1H), 8.02-8.25 (m, 2H), 7.35-7.80 (m, 8H), 6.75-7.00 (m, 3H), 4.30-4.50 (m, 2H), 3.70-3.82 (m, 2H), 3.60-3.70 (m, 8H).
Example 34 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 34-1 in place of the amine starting material. MS (ESI) calculated for C30H24N8O3: 544.20 m/z, found 545.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.19 (s, 1H), 8.41-8.50 (m, 1H), 8.32-8.39 (m, 1H), 7.98-8.12 (m, 2H), 7.81 (s, 1H), 7.60-7.72 (m, 2H), 7.49-7.51 (m, 2H), 7.39-7.42 (m, 2H), 6.85-7.01 (m, 2H), 6.69-6.81 (m, 1H), 6.55 (s, 1H), 4.49 (s, 2H), 3.51-3.61 (m, 2H).
To a solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-chloroimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 1-1) (500 mg, 1.11 mmol, 1 equiv) and pyrazole (151 mg, 2.22 mmol, 2 equiv) in N,N-dimethylformamide (10 mL) were added copper (7.0 mg, 0.11 mmol, 0.1 equiv) and cesium carbonate (1.08 g, 3.33 mmol, 3 equiv). After stirring overnight at 100° C. under a nitrogen atmosphere, the resulting mixture was poured into water giving a suspension. The solid was filtered, dried under air, and purified by silica gel column chromatography using a 0-20% gradient of ethyl acetate in petroleum ether to afford tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-(pyrazol-1-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (230 mg, 41%) as a yellow solid. MS (ESI) calculated for C26H26N8O2: 482.22 m/z, found 483.57 [M+H]+.
tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-(pyrazol-1-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (230 mg, 0.477 mmol, 1 equiv) was dissolved in 4N hydrochloric acid in 1,4-dioxane and the mixture was stirred at room temperature for 1 h. The mixture was concentrated in vacuo to afford 3-{3-[4-(aminomethyl)phenyl]-5-(pyrazol-1-yl)imidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 35-1) (163 mg, 74%) as a yellow solid, which was used without purification in subsequent transformations. MS (ESI) calculated for C21H18N8: 382.17 m/z, found 383.20 [M+H]+.
Example 36 was prepared in a manner analogous to Example 16 using Intermediate 36-1 in place of Intermediate 16-2. MS (ESI) calculated for C34H30N6O2, 554.24 m/z, found 555.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.12 (s, 1H), 8.25-8.27 (m, 1H), 7.97-8.02 (m, 4H), 7.56-7.58 (m, 1H), 7.39-7.58 (m, 7H), 7.18-7.36 (m, 1H), 6.83-6.85 (m, 2H), 6.37-6.40 (m, 1H), 2.80-2.85 (m, 8H).
Intermediate 36-1 was prepared in a manner analogous to Intermediate 11-1 using tert-butyl N-[2-(4-aminophenyl)ethyl]carbamate in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C25H22N6: 406.19 m/z, found 407.05 [M+H]+.
Example 37 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 37-1 in place of Intermediate 1-4. MS (ESI) calculated for C34H28N6O3: 568.22 m/z, found 569.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.72 (s, 1H), 10.20 (s, 1H), 8.66-8.70 (m, 1H), 8.25-8.30 (m, 1H), 7.95-8.05 (m, 4H), 7.60-7.65 (m, 1H), 7.30-7.50 (m, 7H), 7.15-7.25 (m, 1H), 6.94-7.05 (m, 4H), 6.37-6.42 (m, 1H), 4.35-4.40 (m, 2H), 3.70-3.75 (m, 1H), 1.37-1.40 (m, 3H).
To a solution of 2-(4-bromo-2-((4-methoxybenzyl)oxy)phenyl)-1,3-dioxolane (Intermediate 1-3) (4.15 g, 11.3 mmol, 1 equiv) and tert-butyldimethylsilyl methyl carbonate (8.65 g, 45.4 mmol, 4 equiv) in N,N-dimethylformamide (20 mL) were added bis(tri-tert-butylphosphine)palladium(0) (0.60 g, 1.14 mmol, 0.1 equiv) and lithium fluoride (0.6 g, 23 mmol, 2 equiv). The resulting mixture was stirred at 100° C. under nitrogen atmosphere for 2 h then cooled to 0° C. and quenched with water (100 mL). The resulting mixture was extracted with ethyl acetate (200 mL×3). The combined organic layers were washed with water (100 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-60% gradient of ethyl acetate in petroleum ether to provide methyl 2-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl)acetate (3.0 g, 74%) as a light-yellow oil. MS (ESI) calculated for C20H22O6: 358.14 m/z, found 359.10 [M+H]+.
To a cooled (0° C.) solution of methyl 2-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]acetate (500 mg, 1.40 mmol, 1 equiv) in tetrahydrofuran (5 mL) was added sodium hydride (50 mg, 2.1 mmol, 1.5 equiv) and the resulting mixture was stirred at 0° C. for 30 minutes. Methyl iodide (297 mg, 2.09 mmol, 1.5 equiv) was added dropwise at 0° C. and the resulting mixture was warmed to room temperature and stirred for 2 h. The reaction was quenched by addition of 1M hydrochloric acid until the pH was between 5 and 6. The mixture was extracted with ethyl acetate and the combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by silica gel column chromatography using a 0-30% gradient of ethyl acetate in petroleum ether to afford methyl 2-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]propanoate as a yellow oil. MS (ESI) calculated for C21H24O6: 372.16 m/z, found 373.10 [M+H]+.
To a stirred solution of methyl 2-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]propanoate (120 mg, 0.322 mmol, 1 equiv) in tetrahydrofuran (3 mL) and methanol (3 mL) was added a solution of lithium hydroxide (15 mg, 0.64 mmol, 2 equiv) in water (3 mL). The resulting mixture was stirred for 2 h at room temperature. The mixture was acidified to pH 5-6 with 1M hydrochloric acid. The resulting mixture was extracted with ethyl acetate (3×50 mL) and the combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to provide 2-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]propanoic acid (Intermediate 37-1) (100 mg, 87%) as a yellow oil, which was used in subsequent transformations without further purification. MS (ESI) calculated for C20H22O6: 358.14 m/z, found 359.15 [M+H]+.
Example 38 was prepared in a manner analogous to Example 19 using Intermediate 38-1 in place of Intermediate 19-2. MS (ESI) calculated for C34H29N702: 567.20 m/z, found 568.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 10.18 (s, 1H), 8.20-8.25 (m, 1H), 7.92-8.03 (m, 4H), 7.60-7.64 (m, 1H), 7.37-7.50 (m, 3H), 7.19-7.28 (m, 3H), 6.94-7.03 (m, 2H), 6.49-6.55 (m, 2H), 6.41-6.45 (m, 1H), 4.02-4.08 (m, 2H), 3.66-3.74 (m, 3H), 3.55-3.60 (m, 2H).
A suspension of 3-[3-(4-bromophenyl)-5-phenylimidazo[4,5-b]pyridin-2-yl]pyridin-2-amine (Intermediate 29-1) (4.00 g, 9.04 mmol, 1 equiv) and phenyl formate (4.42 g, 36.2 mmol, 4 equiv) in toluene (70 mL) was stirred at 120° C. for 16 h. After concentration, the residue was purified by silica gel column chromatography using a 0-5% gradient of methanol in dichloromethane to afford N-{3-[3-(4-bromophenyl)-5-phenylimidazo[4,5-b]pyridin-2-yl]pyridin-2-yl}formamide (1.9 g, 445%) as a yellow solid. MS (ESI) calculated for C24H16BrN5O: 469.05 m/z, found 470.05, 472.05 [M+H, M+2+H]+.
A suspension of N-{3-[3-(4-bromophenyl)-5-phenylimidazo[4,5-b]pyridin-2-yl]pyridin-2-yl}formamide (169 mg, 0.359 mmol, 1 equiv), tert-butyl N-(azetidin-3-yl)carbamate (124 mg, 0.718 mmol, 2 equiv), EPhos (19 mg, 0.036 mmol, 0.1 equiv), EPhos Pd G4 (33 mg, 0.036 mmol, 0.1 equiv) and cesium carbonate (234 mg, 0.718 mmol, 2 equiv) in 1,4-dioxane (5 mL) was stirred at 120° C. for 2 h under nitrogen atmosphere. The reaction mixture was cooled to room temperature and quenched with water (20 mL). The resulting mixture was extracted with ethyl acetate (20 mL×3) and the combined organic phases were washed with brine (30 mL×3), dried with sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-85% gradient of ethyl acetate in dichloromethane to afford tert-butyl N-(1-{4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]phenyl}azetidin-3-yl)carbamate (156 mg, 62%) as a yellow solid. MS (ESI) calculated for C31H31N7O2: 533.25 m/z, found 534.20 [M+H]+.
A mixture of tert-butyl N-(1-{4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]phenyl}azetidin-3-yl)carbamate (150 mg, 0.281 mmol, 1 equiv) in 2,2,2-trifluoroacetic acid (1 mL) and dichloromethane (5 mL) was stirred at room temperature for 2 h. The mixture was concentrated in vacuo and dissolved in dichloromethane (20 mL). The pH of the mixture was brought to -9 with triethylamine. The mixture was concentrated in vacuo and the resulting residue was purified by silica gel column chromatography using a 0-20% gradient of methanol in dichloromethane to afford 3-{3-[4-(3-aminoazetidin-1-yl)phenyl]-5-phenylimidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (130 mg, 98%) as a light-yellow solid. MS (ESI) calculated for C26H23N7: 433.20 m/z, found 434.15 [M+H]+.
Example 39 was prepared in a manner analogous to Example 19 using Intermediate 39-2 in place of Intermediate 19-2. MS (ESI) calculated for C34H30N6O2: 554.24 m/z, found 555.30 [M+H]+. 1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.28 (s, 1H), 8.32-8.34 (m, 1H), 8.02-8.05 (m, 4H), 7.77 (s, 1H), 7.50-7.53 (m, 1H), 7.47-7.49 (m, 7H), 7.15-7.20 (m, 2H), 6.65-6.75 (m, 1H), 4.31-4.49 (m, 2H), 3.62-3.64 (m, 2H), 3.11-3.30 (m, 2H), 2.80 (s, 3H).
Intermediate 39-2 was prepared in a manner analogous to Intermediate 11-1 using Intermediate 39-1 in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C26H24N6: 420.21 m/z, found 421.15 [M+H]+.
A solution of methyl[2-(4-nitrophenyl)ethyl]amine (5.00 g, 27.7 mmol, 1 equiv), di-tert-butyl dicarbonate (7.56 g, 34.6 mmol, 1.25 equiv) and triethylamine (7.02 g, 69.4 mmol, 2.5 equiv) in dichloromethane (250 mL) was stirred overnight at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was diluted with water and extracted with dichloromethane (50 mL×3). The combined organic layers were washed with water (50 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to afford tert-butyl N-methyl-N-[2-(4-nitrophenyl)ethyl]carbamate (6 g, 71%) as a yellow oil, which was used in the next step without further purification. MS (ESI) calculated for C14H20N2O4: 280.14 m/z, found 281.20 [M+H]+.
A mixture of tert-butyl N-methyl-N-[2-(4-nitrophenyl)ethyl]carbamate (5.00 g, 17.8 mmol, 1 equiv) and 10% palladium on carbon (1.90 g, 17.8 mmol, 1 equiv) in methanol (50 mL) and ethyl acetate (100 mL) was stirred overnight at room temperature under hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with ethyl acetate/methanol (200 mL). The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica gel column chromatography using a 0-100% gradient of ethyl acetate in petroleum ether to afford tert-butyl N-[2-(4-aminophenyl)ethyl]-N-methylcarbamate (Intermediate 39-1) (2.5 g, 60%) as a red oil. MS (ESI) calculated for C4H22N2O2: 250.17 m/z, found 251.30 [M+H]+.
Example 40 was prepared in a manner analogous to Example 19 using Intermediate 36-1 in place of Intermediate 19-2 and Intermediate 40-1 in place of Intermediate 19-3. MS (ESI) calculated for C32H26FN702: 558.21 m/z, found 559.22 [M+H]+. 1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.17 (s, 1H), 8.25-8.27 (m, 1H), 7.96-8.02 (m, 4H), 7.37-7.47 (m, 8H), 7.17-7.19 (m, 1H), 6.87-6.94 (m, 1H), 6.37-6.39 (m, 1H), 3.72-3.82 (m, 2H), 2.85 (s, 4H). 19F-NMR (400 MHz, DMSO-d6) δ (ppm): −141.32.
A mixture of 2-{4-bromo-3-fluoro-2-[(4-methoxyphenyl)methoxy]phenyl}-1,3-dioxolane (Intermediate 21-1) (1.00 g, 2.61 mmol, 1 equiv), 2-ethenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.01 g, 13.1 mmol, 5 equiv), [1,1′-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (170 mg, 0.26 mmol, 0.1 equiv) and potassium carbonate (1.08 g, 7.83 mmol, 3 equiv) in tetrahydrofuran (8 mL) and water (2 mL) was stirred for 3 h at 80° C. under nitrogen atmosphere. The mixture cooled to room temperature and quenched with water (100 mL). The mixture was extracted with ethyl acetate (3×100 mL) and the combined organic phases were concentrated under reduced pressure. The residue was purified by reverse-phase flash column chromatography on C18 silica gel using a 10-50% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) to afford 2-{4-ethenyl-3-fluoro-2-[(4-methoxyphenyl)methoxy]phenyl}-1,3-dioxolane (800 mg, 92%) as a yellow oil. MS (ESI) calculated for C19H19FO4: 330.13 m/z, found 331.13 [M+H]+.
A mixture of 2-{4-ethenyl-3-fluoro-2-[(4-methoxyphenyl)methoxy]phenyl}-1,3-dioxolane (260 mg, 0.787 mmol, 1 equiv), osmium tetroxide (0.30 mL, 5.79 mmol, 7.4 equiv) and sodium periodate (505 mg, 2.36 mmol, 3 equiv) in tetrahydrofuran (10 mL) and water (10 mL) was stirred for 20 min at room temperature. The reaction was quenched with water (50 mL) and the mixture was extracted with ethyl acetate (3×40 mL). The combined organic phases were dried with sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (eluting with petroleum ether/ethyl acetate (10:1)) to afford 4-(1,3-dioxolan-2-yl)-2-fluoro-3-[(4-methoxyphenyl)methoxy]benzaldehyde (Intermediate 40-1) (150 mg, 57%) as a white solid. MS (ESI) calculated for C18H17FO5: 332.11 m/z, found 333.11 [M+H]+.
Example 41 was prepared in manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 41-1 in place of Intermediate 1-4. MS (ESI) calculated for C36H32N6O3: 596.25 m/z, found 597.1 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm) 12.08 (s, 1H), 10.04 (s, 1H), 9.10-9.34 (m, 1H), 8.27-8.36 (m, 2H), 7.90-8.20 (m, 5H), 7.69-7.98 (m, 2H), 7.29-7.51 (m, 7H), 6.64-6.75 (m, 1H), 4.62 (d, J=5.9 Hz, 2H), 1.42 (s, 9H).
To a stirred solution of 3-tert-butyl-5-formyl-4-hydroxybenzoic acid (250 mg, 1.13 mmol, 1 equiv), p-methoxybenzyl chloride (210 mg, 1.35 mmol, 1.2 equiv) and potassium carbonate (621 mg, 4.50 mmol, 4 equiv) in anhydrous acetonitrile (5 mL) was added potassium iodide (93 mg, 0.56 mmol, 0.5 equiv) and the resulting mixture was stirred at 50° C. for 5 h. Water (10 mL) was added and the mixture was extracted with ethyl acetate (10 mL×3). The combined organic layers were washed with brine (30 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-50% gradient of ethyl acetate in petroleum ether to afford (4-methoxyphenyl)methyl 3-tert-butyl-5-formyl-4-[(4-methoxyphenyl)methoxy]benzoate (260 mg, 50%) as a yellow solid. MS (ESI) calculated for C28H30O6: 462.20 m/z, found 463.20 [M+H]+.
To a stirred solution of (4-methoxyphenyl)methyl 3-tert-butyl-5-formyl-4-[(4-methoxyphenyl)methoxy]benzoate (280 mg, 0.605 mmol, 1 equiv) in tetrahydrofuran (5 mL) was added a solution of lithium hydroxide (87 mg, 3.6 mmol, 6 equiv) in water (4 mL) and the resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated in vacuo and purified by reverse-phase column chromatography on C18 silica gel using a 0-95% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) to afford 3-tert-butyl-5-formyl-4-[(4-methoxyphenyl)methoxy]benzoic acid (Intermediate 41-1) (200 mg, 96%) as a yellow solid. MS (ESI) calculated for C20H22O5: 342.15 m/z, found 341.2 [M+H]+.
Example 42 was prepared in a manner analogous to Example 1 using Intermediate 42-2 in place of 4-(morpholin-4-yl)phenylboronic acid. MS (ESI) calculated for C42H39FN8O4: 738.31 m/z, found 739.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.22 (s, 1H), 8.30-8.35 (m, 1H), 8.00-8.10 (m, 1H), 7.75-7.85 (m, 1H), 7.65-7.75 (m, 1H), 7.55-7.62 (m, 1H), 7.40-7.50 (m, 2H), 7.30-7.40 (m, 2H), 7.20-7.30 (m, 1H), 7.15-7.20 (m, 2H), 6.95 (s, 1H), 6.80-6.95 (m, 1H), 6.70-6.80 (m, 1H), 4.25-4.40 (m, 4H), 3.85-4.00 (m, 1H), 3.55 (s, 2H), 3.30-3.40 (m, 2H), 2.70-2.90 (m, 2H), 2.20-2.30 (m, 2H), 1.80-2.00 (m, 4H), 1.60-1.70 (m, 2H).
To a mixture of 1-(piperidin-4-yl)pyrrolidin-2-one (1.00 g, 5.94 mmol, 1 equiv), 1,3-dibromo-2-fluorobenzene (1.51 g, 5.94 mmol, 1 equiv) and sodium tert-butoxide (1.71 g, 17.8 mmol, 3 equiv) in toluene (10 mL) were added tris(dibenzylideneacetone)dipalladium(0) (0.54 g, 0.59 mmol, 0.1 equiv) and BINAP (0.74 g, 1.2 mmol, 0.2 equiv). The resulting mixture was stirred overnight at 120° C. under nitrogen atmosphere. The mixture was then cooled to room temperature, concentrated under reduced pressure, and purified by silica gel column chromatography using a 0-5% gradient of methanol in dichloromethane to afford 1-[1-(3-bromo-2-fluorophenyl)piperidin-4-yl]pyrrolidin-2-one (860 mg, 31%) as a brown oil. MS (ESI) calculated for C15H18BrFN2O: 340.06 m/z, found 341.00 [M+H]+.
To a solution of 1-[1-(3-bromo-2-fluorophenyl)piperidin-4-yl]pyrrolidin-2-one (860 mg, 2.52 mmol, 1 equiv) and bis(pinacolato)diboron (448 mg, 1.76 mmol, 0.7 equiv) in 1,4-dioxane (10 mL) were added potassium acetate (742 mg, 7.56 mmol, 3 equiv) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (184 mg, 0.252 mmol, 0.1 equiv). The resulting mixture was stirred overnight at 120° C. under a nitrogen atmosphere, cooled to room temperature and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography on C18 silica gel using a 10-50% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) to provide 2-fluoro-3-[4-(2-oxopyrrolidin-1-yl)piperidin-1-yl]phenylboronic acid (Intermediate 42-1) (180 mg, 20%) as a white oil. MS (ESI) calculated for C15H20BFN2O3: 306.16 m/z, found 307.10 [M+H]+.
To a suspension of 3-(3-(4-(aminomethyl)phenyl)-3H-imidazo[4,5-b]pyridin-2-yl)pyridin-2-amine (Intermediate 74-1) (120 mg, 231 μmol, 1 equiv), HATU (108 mg, 278 μmol, 1.2 equiv) and 2-(1-(dimethylcarbamoyl)-1H-indol-6-yl)acetic acid (Intermediate 43-1) (70.4 mg, 243 μmol, 1.05 equiv) in N,N-dimethylformamide (1.9 mL) was added N,N-diisopropylethylamine (162 μL, 925 μmol, 4 equiv) and the mixture was stirred at room temperature for 5 min. The reaction mixture was then diluted with dichloromethane (10 mL) and water (10 mL). The layers were separated, and the aqueous layer was extracted with dichloromethane (10 mL). The organic layers were combined, dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography using a 0-10% gradient of methanol in dichloromethane to give 6-(2-((4-(2-(2-aminopyridin-3-yl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)amino)-2-oxoethyl)-N,N-dimethyl-1H-indole-1-carboxamide (78.5 mg, 62%) as a yellow oil. MS (ESI) calculated for C31H28N8O2: 544.23 m/z, found 545.42 [M+H]+.
To a solution of 6-(2-((4-(2-(2-aminopyridin-3-yl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)amino)-2-oxoethyl)-N,N-dimethyl-1H-indole-1-carboxamide (60 mg, 110 μmol), in dichloromethane (5 mL) and methanol (500 μL) was added trifluoroacetic acid (17 μL, 220 μmol, 2 equiv) and the mixture was cooled to −78° C. Ozone was then bubbled through the solution for 5 min. The reaction mixture was sparged with nitrogen for 5 min, then quenched with dimethyl sulfide (16.3 μL, 220 μmol, 2 equiv). The mixture was allowed to warm to 0° C. and stirring was continued for 1 h. The reaction mixture was then concentrated in vacuo, coevaporating with dichloromethane (3×10 mL). Potassium carbonate (60.9 mg, 441 μmol, 4 equiv) and methanol (5 mL) were added to the residue and the mixture was stirred at room temperature overnight. The reaction mixture was quenched with 1M hydrochloric acid (5 mL), and the mixture was concentrated in vacuo. The residue was purified by Preparative HPLC on a CSH C18 column using a 10-30% gradient of acetonitrile in water (+0.1% formic acid) to give N-(4-(2-(2-aminopyridin-3-yl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)-2-(3-(3,3-dimethylureido)-4-formylphenyl) acetamide (Example 43) (20.2 mg, 33%) as a white solid. MS (ESI) calculated for C30H28N8O3: 548.23 m/z, found 549.41 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.64 (s, 1H), 9.92 (s, 1H), 8.75 (t, J=5.9 Hz, 1H), 8.43 (s, 1H), 8.31 (dd, J=4.7, 1.3 Hz, 1H), 8.20 (dd, J=8.0, 1.3 Hz, 1H), 7.98 (dd, J=4.8, 1.8 Hz, 1H), 7.78 (d, J=7.9 Hz, 1H), 7.44-7.33 (m, 5H), 7.21 (dd, J=7.7, 1.7 Hz, 1H), 7.10 (dd, J=7.9, 1.2 Hz, 1H), 6.97 (s, 2H), 6.39 (dd, J=7.7, 4.8 Hz, 1H), 4.37 (d, J=5.9 Hz, 2H), 3.58 (s, 2H), 2.98 (s, 6H).
To a cooled (0° C.) suspension of sodium hydride (240 mg, 6.00 mmol, 1.2 equiv) in tetrahydrofuran (6 mL) was added a solution of 6-bromoindole (1.00 g, 5.00 mmol, 1 equiv) in tetrahydrofuran (4 mL) and the resulting dark brown solution was stirred at 0° C. for 5 min. The mixture was then stirred at room temperature for 1 h then cooled to 0° C. Dimethylcarbamoyl chloride (517 μL, 5.50 mmol, 1.1 equiv) was added dropwise and the resulting mixture was stirred overnight at room temperature. The reaction was then quenched with saturated aqueous ammonium chloride (10 mL) and water (10 mL) and diluted with ethyl acetate (20 mL). The layers were separated, and the aqueous layer was extracted with ethyl acetate (10 mL). The organic layers were combined, washed with brine (20 mL), then dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography using a 0-25% gradient of ethyl acetate in hexanes to give 6-bromo-N,N-dimethyl-1H-indole-1-carboxamide (1.12 g, 84%) as a red oil. MS (ESI) calculated for C11H11BrN2O: 266.01 m/z, found 267.06, 269.04 [M+H, M+2+H]+.
To a solution of 6-bromo-N,N-dimethyl-1H-indole-1-carboxamide (1.16 g, 4.34 mmol, 1 equiv) in tetrahydrofuran (8.69 mL) was added QPhos (157 mg, 217 μmol, 0.05 equiv) and tris(dibenzylideneacetone)dipalladium(0) (203 mg, 217 μmol, 0.05 equiv) and the resulting solution was sparged with nitrogen for 5 mins. 2-(tert-butoxy)-2-oxoethylzinc chloride (0.5 M in diethyl ether, 13.0 mL, 6.51 mmol, 1.5 equiv) was then added to the solution and the mixture was stirred at room temperature for 3 h under a nitrogen atmosphere. The reaction mixture was then concentrated in vacuo to give tert-butyl 2-(1-(dimethylcarbamoyl)-1H-indol-6-yl)acetate as a red oil, which was used directly in the next step without purification. MS (ESI) calculated for C7H22N2O3: 302.16 m/z, found 301.11 [M−H]−.
To a solution of crude tert-butyl 2-(1-(dimethylcarbamoyl)-1H-indol-6-yl)acetate (1.31 g, 4.34 mmol) in dichloromethane (17 mL) was added trifluoroacetic acid (10.1 mL) and the mixture was stirred for 10 min at room temperature. The reaction mixture was then concentrated in vacuo, co-evaporating with dichloromethane (2×20 mL). The residue was then redissolved in dichloromethane (20 mL) and extracted with 2M aqueous sodium hydroxide (50 mL). The aqueous layer was then acidified with 6M aqueous hydrochloric acid (15 mL) and extracted with ethyl acetate (2×30 mL). The organic layers were combined, dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography using a 0-5% gradient of methanol in dichloromethane to give 2-(1-(dimethylcarbamoyl)-1H-indol-6-yl)acetic acid (Intermediate 43-1) (327 mg, 31% over two steps) as a yellow oil. MS (ESI) calculated for C13H14N2O3: 246.10 m/z, found 247.15 [M+H]+.
Example 44 was prepared in a manner analogous to Example 43 using Intermediate 44-2 in place of Intermediate 43-1. MS (ESI) calculated for C32H27N903: 585.22 m/z, found 585.83 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 11.33 (s, 1H), 9.97 (s, 1H), 8.78 (t, J=5.9 Hz, 1H), 8.36 (s, 1H), 8.32 (s, 1H), 8.30 (dd, J=4.8, 1.4 Hz, 1H), 8.20 (dd, J=8.0, 1.4 Hz, 1H), 7.98 (dd, J=4.8, 1.8 Hz, 1H), 7.93 (s, 1H), 7.86 (d, J=7.9 Hz, 1H), 7.43-7.33 (m, 5H), 7.26 (dd, J=7.9, 1.3 Hz, 1H), 7.22 (dd, J=7.6, 1.8 Hz, 1H), 6.98 (s, 2H), 6.39 (dd, J=7.7, 4.9 Hz, 1H), 4.38 (d, J=5.9 Hz, 2H), 3.92 (s, 3H), 3.64 (s, 2H).
Intermediate 44-2 was prepared in a manner analogous to Intermediate 43-1 (starting from Step 2) using Intermediate 44-1 in place of the bromide starting material. MS (ESI) calculated for C15H13N3O3: 283.10 m/z, found 283.86 [M+H]+.
1-Methyl-1H-pyrazole-4-carboxylic acid (750 mg, 5.83 mmol) was dissolved in thionyl chloride (2.56 mL, 35.0 mmol, 6 equiv) and the mixture was heated to 80° C. for 3 h. The reaction mixture was then concentrated in vacuo to give 1-methyl-1H-pyrazole-4-carbonyl chloride (842 mg, crude quant) as a brown solid, which was used directly in the next step without further purification.
To a solution of 6-iodo-1H-indole (1.13 g, 4.65 mmol, 1 equiv) and 4-dimethylaminopyridine (58.0 mg, 465 μmol, 0.1 equiv) in dichloromethane (18.6 mL) was added triethylamine (977 μL, 6.97 mmol, 1.5 equiv) and the solution was cooled to 0° C. 1-methyl-1H-pyrazole-4-carbonyl chloride (840 mg, 5.81 mmol, 1.25 equiv) was added to the mixture and the reaction was slowly allowed to reach room temperature and stirred overnight. The reaction mixture was then washed with saturated aqueous ammonium chloride (2×15 mL) and brine (15 mL), then dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography using a 30-100% gradient of ethyl acetate in hexanes to give (6-iodo-1H-indol-1-yl)(1-methyl-1H-pyrazol-4-yl)-methanone (Intermediate 44-1) (1.39 g, 85% over two steps) as a brown solid. MS (ESI) calculated for C13H10IN3O: 350.99 m/z, found 352.15 [M+H]+.
Example 45 was prepared in a manner analogous to Example 43 using furo[2,3-c]pyridine-5-carboxylic acid in place of Intermediate 43-1. MS (ESI) calculated for C25H19N7O3: 465.15 m/z, found 465.80 [M+H]+. 1H NMR (400 MHz, methanol-d4) δ (ppm): 10.15 (s, 1H), 8.61 (s, 1H), 8.50 (dd, J=4.9, 1.3 Hz, 1H), 8.46 (s, 1H), 8.33 (dt, J=17.2, 8.6 Hz, 1H), 8.12 (dt, J=13.2, 6.6 Hz, 1H), 7.77 (dt, J=10.7, 5.3 Hz, 1H), 7.64 (d, J=8.3 Hz, 2H), 7.55-7.45 (m, 3H), 6.75 (dt, J=24.4, 12.2 Hz, 1H), 4.83 (s, 2H).
Example 46 was prepared in a manner analogous to Example 43 using furo[2,3-b]pyridine-5-carboxylic acid in place of Intermediate 43-1 and omitting the potassium carbonate/methanol step. MS (ESI) calculated for C25H19N7O3: 465.15 m/z, found 465.85 [M+H]+. 1H NMR (400 MHz, methanol-d4) δ (ppm): 10.18 (s, 1H), 8.74-8.68 (m, 1H), 8.67-8.59 (m, 1H), 8.47 (t, J=10.3 Hz, 1H), 8.31 (d, J=11.5 Hz, 1H), 8.10 (d, J=5.6 Hz, 1H), 7.76 (d, J=7.4 Hz, 1H), 7.68-7.58 (m, 2H), 7.54-7.44 (m, 3H), 6.81-6.70 (m, 1H), 4.77 (s, 2H).
Example 47 was prepared in a manner analogous to Example 43 using furo[3,2-b]pyridine-5-carboxylic acid in place of Intermediate 43-1 and omitting the potassium carbonate/methanol step. MS (ESI) calculated for C25H19N7O3: 465.15 m/z, found 465.85 [M+H]+. 1H NMR (400 MHz, methanol-d4) δ (ppm): 10.01 (d, J=18.4 Hz, 1H), 8.47 (dt, J=8.1, 4.0 Hz, 1H), 8.39 (d, J=7.4 Hz, 1H), 8.31 (dt, J=14.9, 7.5 Hz, 1H), 8.09 (dt, J=13.6, 6.7 Hz, 1H), 7.75 (dt, J=6.0, 3.0 Hz, 1H), 7.62 (d, J=8.4 Hz, 2H), 7.57 (dd, J=11.0, 5.9 Hz, 1H), 7.53-7.43 (m, 3H), 6.77-6.69 (m, 1H), 4.81 (d, J=9.9 Hz, 2H).
Example 48 was prepared in a manner analogous to Example 43 using Intermediate 48-2 in place of Intermediate 43-1 and EDCI (2 equiv)/pyridine (20 equiv)/48 h in place of HATU/N,N-diisopropylethylamine/5 min. MS (ESI) calculated for C31H27N7O3: 545.22 m/z, found 546.39 [M+H]+. 1H NMR (400 MHz, methanol-d4) δ (ppm): 9.94 (s, 1H), 8.54 (s, 1H), 8.31 (dd, J=10.7, 9.7 Hz, 1H), 8.18 (dt, J=17.0, 8.4 Hz, 1H), 8.00-7.90 (m, 1H), 7.79 (d, J=7.9 Hz, 1H), 7.52-7.39 (m, 3H), 7.36 (d, J=8.3 Hz, 2H), 7.32-7.19 (m, 2H), 6.41 (dt, J=24.6, 12.3 Hz, 1H), 4.48 (s, 2H), 3.66 (s, 2H), 1.74 (ddd, J=12.5, 7.7, 4.6 Hz, 1H), 1.02-0.85 (m, 4H).
Intermediate 48-2 was prepared in a manner analogous to Intermediate 43-1 (starting from Step 2) using Intermediate 48-1 in place of the bromide starting material. MS (ESI) calculated for C4H13NO3: 243.09 m/z, found 244.14 [M+H]+.
Step 1: Synthesis of (6-bromo-1H-indol-1-yl)(cyclopropyl)methanone (Intermediate 48-1)
To a solution of 6-bromoindole (500 mg, 2.50 mmol, 1 equiv) and 4-dimethylaminopyridine (31.2 mg, 250 μmol, 1 equiv) in dichloromethane (10.0 mL) was added triethylamine (525 μL, 3.75 mmol, 1.5 equiv) and the solution was cooled to 0° C. Cyclopropanecarbonyl chloride (289 μL, 3.12 mmol, 1.25 equiv) was then added and the reaction was allowed to warm to room temperature and stirred for 48 h. The reaction was then quenched with saturated aqueous ammonium chloride (10 mL) and water (10 mL) and diluted with dichloromethane (20 mL). The layers were separated, and the organic layer was washed again with saturated aqueous ammonium chloride (20 mL). The organic layer was then dried over sodium sulfate, filtered, and concentrated in vacuo to give (6-bromo-1H-indol-1-yl)(cyclopropyl)-methanone (Intermediate 48-1) (660 mg, crude) as a brown solid, which was used in subsequent transformations without further purification. MS (ESI) calculated for C12H10BrNO: 262.99 m/z, found 261.88, 262.88 [M−H]−.
Example 49 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 3-1 in place of the amine starting material. MS (ESI) calculated for C31H29N7O4: 563.23 m/z, found 564.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.18 (s, 1H), 7.93-8.00 (m, 2H), 7.57-7.60 (m, 1H), 7.28-7.36 (m, 4H), 7.03-7.06 (m, 1H), 6.84-6.91 (m, 3H), 6.34-6.38 (m, 1H), 4.37 (s, 2H), 3.64-3.72 (m, 6H), 3.39-3.41 (m, 4H).
To a solution of 3-bromo-4-hydroxybenzoic acid (2.00 g, 9.22 mmol, 1 equiv) in 2,2,2-trifluoroacetic acid (5 mL) was added a solution of 1,3,5,7-tetraazaadamantane (3.59 g, 25.6 mmol, 2 equiv) in 2,2,2-trifluoroacetic acid (5 mL) dropwise and the obtained solution was stirred at 90° C. for 23 h. After cooling to room temperature, 150 mL of water was added, and the resulted mixture was acidified with 4N aqueous hydrochloric acid giving a precipitate. The precipitate was collected by filtration, washed with water, and dried in an oven to provide 3-bromo-5-formyl-4-hydroxybenzoic acid (2.5 g, 96%) as a brown solid, which was used directly in the next step without further purification. MS (ESI) calculated for C8H5BrO4: 243.94 m/z, found 246.90 [M+H+2]+.
A solution of 3-bromo-5-formyl-4-hydroxybenzoic acid (187 mg, 0.765 mmol, 1.5 equiv), 3-{3-[4-(aminomethyl)phenyl]-5-phenylimidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 4-1) (200 mg, 0.510 mmol, 1.00 equiv), N,N-diisopropylethylamine (263 mg, 2.04 mmol, 4 equiv) and HATU (291 mg, 0.765 mmol, 1.5 equiv) in N,N-dimethylformamide (5 mL) was stirred at room temperature for 2 h. The solution was purified by preparative HPLC on a XBridge Prep OBD C18 Column using a 17-42% gradient of acetonitrile in water (+10 mmol/L ammonium bicarbonate) to provide N-({4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)-3-bromo-5-formyl-4-hydroxybenzamide (Example 50) (8.3 mg, 2%) as a light-yellow solid. MS (ESI) calculated for C32H23BrN6O3: 618.10 m/z, found 618.95, 620.95 [M+H, M+2+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.13 (s, 1H), 9.43 (s, 1H), 8.23-8.56 (m, 1H), 7.92-8.82 (m, 6H), 7.36-7.55 (m, 8H), 7.21-7.24 (m, 1H), 4.53 (s, 2H).
Example 51 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 51-2 in place of the amine starting material. MS (ESI) calculated for C32H28N6O4: 560.22 m/z, found 561.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.18 (s, 1H), 8.20-8.23 (m, 1H), 7.62-8.04 (m, 1H), 7.41-7.60 (m, 7H), 6.90-6.95 (m, 2H), 6.65-6.69 (m, 1H), 4.37 (s, 2H), 3.55 (s, 2H), 1.54 (s, 6H).
Intermediate 51-2 was prepared in a manner analogous to Intermediate 11-1 using Intermediate 51-1 in place of 2-chloro-3-nitro-6-phenylpyridine and tert-butyl N-[(4-aminophenyl)methyl]carbamate in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C23H22N6O: 398.19 m/z, found 399.13 [M+H]+.
To a stirred solution of 6-bromo-2-chloro-3-nitropyridine (1.00 g, 4.21 mmol, 1 equiv) in anhydrous N,N-dimethylformamide (14 mL) were added 2-methyl-3-butyn-2-ol (0.53 g, 6.3 mmol, 1.5 equiv), copper(I) iodide (0.64 g, 3.4 mmol, 0.8 equiv), bis(triphenylphosphine)palladium(II) dichloride (0.15 g, 0.21 mmol, 0.05 equiv) and N,N-diisopropylethylamine (1.36 g, 10.5 mmol, 2.5 equiv). The mixture was stirred at 80° C. for 7 h under nitrogen atmosphere. Water was added and the resulting mixture was extracted with ethyl acetate (3×60 mL). The combined organic layers were washed with brine (2×40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-95% gradient of ethyl acetate in petroleum ether to afford 4-(6-chloro-5-nitropyridin-2-yl)-2-methylbut-3-yn-2-ol (Intermediate 51-1) (700 mg, 62%) as a brown solid. MS (ESI) calculated for C10H9ClN2O3: 240.03 m/z, found 241.06 [M+H]+.
Example 52 was prepared in a manner analogous to Example 50 using 3-chloro-4-hydroxybenzoic acid in place of 3-bromo-4-hydroxybenzoic acid. MS (ESI) calculated for C32H23ClN6O3: 574.15 m/z, found 575.10 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm) 10.19 (s, 1H), 9.08 (s, 1H), 8.12-8.33 (m, 2H), 7.88-8.12 (m, 5H), 7.34-7.58 (m, 8H), 7.18-7.30 (m, 1H), 6.95 (s, 2H), 6.38-6.52 (m, 1H), 4.45-4.68 (m, 2H).
Example 53 was prepared in a manner analogous to Example 50 using 3-fluoro-4-hydroxybenzoic acid in place of 3-bromo-4-hydroxybenzoic acid. MS (ESI) calculated for C32H23FN6O3: 558.18 m/z, found 559.10 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 10.29 (s, 1H), 9.10-9.21 (m, 1H), 8.26-8.28 (m, 1H), 8.03-8.09 (m, 1H), 7.91-8.01 (m, 5H), 7.40-7.49 (m, 7H), 7.22-7.24 (m, 1H), 6.42-6.45 (m, 1H), 4.57-4.59 (m, 2H). 19F NMR (400 MHz, DMSO-d6) δ-134.54.
Example 54 was prepared in a manner analogous to Example 1 using 2-fluorophenylboronic acid in place of 4-(morpholin-4-yl)phenylboronic acid. MS (ESI) calculated for C33H25FN6O3: 572.20 m/z, found 573.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.18 (s, 1H), 8.28-8.31 (m, 1H), 8.00-8.02 (m, 1H), 7.76-7.78 (m, 2H), 7.59-7.62 (m, 1H), 7.21-7.45 (m, 9H), 6.88-6.95 (m, 2H), 6.41-6.45 (m, 1H), 4.37 (s, 2H), 3.52-3.55 (m, 2H).
Example 55 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 12-1 in place of the amine starting material. MS (ESI) calculated for C37H33N7O4: 639.26 m/z, found 640.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.18 (s, 1H), 8.23-8.26 (m, 1H), 7.97-8.02 (m, 2H), 7.59-7.62 (m, 2H), 7.39-7.48 (m, 5H), 7.29-7.35 (m, 1H), 7.21-7.25 (m, 1H), 6.88-7.01 (m, 3H), 6.41-6.45 (m, 1H), 4.39 (s, 2H), 3.76-3.79 (m, 4H), 3.52 (s, 2H), 3.15-3.18 (m, 4H).
Example 56 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 56-1 in place of Intermediate 1-4. MS (ESI) calculated for C34H27N7O3: 581.22 m/z, found 582.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.11 (s, 1H), 8.31-8.32 (m, 1H), 8.25-8.27 (m, 1H), 7.97-8.02 (m, 5H), 7.34-7.49 (m, 8H), 7.21-7.32 (m, 1H), 6.41-6.44 (m, 1H), 4.32-4.34 (m, 2H), 1.46-1.47 (m, 2H), 1.14-1.15 (m, 2H).
A mixture of methyl 5-bromo-3-hydroxypyridine-2-carboxylate (5.00 g, 21.5 mmol, 1 equiv), p-methoxybenzyl chloride (4.05 g, 25.9 mmol, 1.2 equiv), potassium iodide (0.36 g, 2.2 mmol, 0.1 equiv) and potassium carbonate (8.93 g, 64.6 mmol, 3 equiv) in N,N-dimethylformamide (30 mL) was stirred overnight at 70° C. under nitrogen atmosphere. The reaction was cooled to room temperature, quenched by the addition of water (30 mL), and extracted by ethyl acetate (3×50 mL). The combined organic layers were washed with brine (30 mL), dried with sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-100% gradient of ethyl acetate in petroleum ether to afford methyl 5-bromo-3-[(4-methoxyphenyl)methoxy]pyridine-2-carboxylate (6 g, 67%) as a yellow oil. MS (ESI) calculated for C15H14BrNO4: 351.01 m/z, found 352.10 [M+H]+.
To a cooled (−78° C.) solution of methyl 5-bromo-3-[(4-methoxyphenyl)methoxy]pyndine-2-carboxylate (6.00 g, 17.0 mmol, 1 equiv) in tetrahydrofuran (150 mL) was added dropwise diisobutylaluminum hydride (1M in toluene) (51 mL, 51 mmol, 3 equiv). The obtained mixture was stirred at room temperature for 1 h. The reaction was quenched with aqueous potassium sodium tartrate and then methanol (20 mL). Stirring was continued for 10 min and the mixture was extracted with ethyl acetate (100 mL×3). The combined organic extracts were washed with brine (200 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-100% gradient of ethyl acetate in petroleum ether to afford {5-bromo-3-[(4-methoxyphenyl)methoxy]pyridin-2-yl}methanol (3.5 g, 57%) as a yellow solid. MS (ESI) calculated for C14H14BrNO3: 323.02 m/z, found 324.10 [M+H]+.
To a cooled (0° C.) solution of {5-bromo-3-[(4-methoxyphenyl)methoxy]pyridin-2-yl}methanol (3.50 g, 10.8 mmol, 1 equiv) in dichloromethane (50 mL) was added Dess-Martin periodinane (5.50 g, 13.0 mmol, 1.2 equiv). The mixture was stirred at room temperature for 1 h. The reaction was quenched with water and extracted with ethyl acetate (100 mL×3). The combined organic extracts were washed with brine (100 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography to afford 5-bromo-3-[(4-methoxyphenyl)methoxy]pyridine-2-carbaldehyde (1.6 g, 41%) as a yellow solid. MS (ESI) calculated for C14H12BrNO3: 321.00 m/z, found 320.10 [M−H]−.
A mixture of 5-bromo-3-((4-methoxybenzyl)oxy)picolinaldehyde (2.70 g, 8.38 mmol, 1 equiv), p-toluenesulfonic acid (0.14 g, 0.84 mmol, 0.1 equiv), ethylene glycol (7.09 g, 114 mmol, 5 equiv) and triethylorthoformate (3.73 g, 25.1 mmol, 3 equiv) in toluene (30 mL) was stirred overnight at 90° C. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography using a 0-100% gradient of ethyl acetate in petroleum ether to afford 5-bromo-2-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)pyridine (1.5 g, 48%) as a white solid. MS (ESI) calculated for C1H16BrNO4: 365.03 m/z, found 366.20 [M+H]+.
A mixture of 5-bromo-2-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]pyridine (1.7 g, 4.6 mmol, 1 equiv), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.34 g, 0.46 mmol, 0.1 equiv) and XPhos (0.44 g, 0.93 mmol, 0.2 equiv) in a solution of (2-ethoxy-2-oxoethyl)zinc(II) bromide (45 mL, 193 mmol) in tetrahydrofuran was stirred overnight at 80° C. under nitrogen atmosphere. The mixture was concentrated and purified by reverse phase flash column chromatography on C18 silica gel using a 5-70% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) to afford ethyl 2-[6-(1,3-dioxolan-2-yl)-5-[(4-methoxyphenyl)methoxy]pyridin-3-yl]acetate (350 mg, 19%) as a yellow solid. MS (ESI) calculated for C20H23NO6, 373.15 m/z, found 374.10 [M+H]+.
To a solution of ethyl 2-[6-(1,3-dioxolan-2-yl)-5-[(4-methoxyphenyl)methoxy]pyridin-3-yl]acetate (350 mg, 0.937 mmol, 1 equiv) in dimethyl sulfoxide (5 mL) were added ethenyl diphenylsulfanium triflate (240 mg, 1.12 mmol, 1.2 equiv) and 1,8-diazabicyclo(5.4.0)undec-7-ene (419 mg, 2.81 mmol, 3 equiv). The resulting mixture was stirred for 6 h at room temperature then diluted with water (10 mL) and extracted with ethyl acetate (20 mL×3). The combined organic phases were washed with brine (50 mL×3), dried with sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-100% gradient of ethyl acetate in petroleum ether to afford ethyl 1-[6-(1,3-dioxolan-2-yl)-5-[(4-methoxyphenyl)methoxy]pyridin-3-yl]cyclopropane-1-carboxylate (300 mg, 80%) as a white solid. MS (ESI) calculated for C22H25NO6: 399.17 m/z, found 400.02 [M+H]+.
To a solution of methyl 1-[6-(1,3-dioxolan-2-yl)-5-[(4-methoxyphenyl)methoxy]pyridin-3-yl]cyclopropane-1-carboxylate (300 mg, 0.778 mmol, 1 equiv) in tetrahydrofuran (3 mL) was added a solution of lithium hydroxide (56 mg, 2.3 mmol, 3 equiv) in water (1 mL). The mixture was stirred at room temperature for 0.5 h. The pH was adjusted to ˜6 with 1N hydrochloric acid and the mixture was concentrated in vacuo to afford 1-(6-(1,3-dioxolan-2-yl)-5-((4-methoxybenzyl)oxy)pyridin-3-yl)cyclopropane-1-carboxylic acid (Intermediate 56-1) as a brown solid, which was used without further purification in subsequent transformations. MS (ESI) calculated for C20H21NO6: 371.14 m/z, found 372.14 [M+H]+.
To a stirred mixture of benzyl N-[2-(trifluoro-lambda4-boranyl)ethyl]carbamate potassium (4.76 g, 16.7 mmol, 3 equiv) in toluene (200 mL) and water (20 mL) were added cesium carbonate (3.63 g, 11.1 mmol, 2 equiv), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.41 g, 0.56 mmol, 0.1 equiv), RuPhos (0.26 g, 0.56 mmol, 0.1 equiv) and 5-bromo-2-(1,3-dioxolan-2-yl)phenoxy(tert-butyl)dimethylsilane (Intermediate 16-3) (2.00 g, 5.57 mmol, 1 equiv) and the resulting mixture was stirred overnight at 80° C. under nitrogen atmosphere. The mixture was cooled to room temperature and diluted with water (20 mL). The resulting mixture was extracted with ethyl acetate (3×150 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-20% gradient of ethyl acetate in petroleum ether to afford benzyl N-(2-{3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)phenyl}ethyl)carbamate (680 mg, 20%) as a yellow viscous oil. MS (ESI) calculated for C25H35NO5Si: 457.23 m/z, found 458.35 [M+H]+.
To a cooled (0° C.) mixture of benzyl N-(2-{3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)phenyl}ethyl)carbamate (680 mg, 1.49 mmol, 1 equiv) in N,N-dimethylformamide (5 mL) was added sodium hydride (75 mg, 3.0 mmol, 2.1 equiv) under nitrogen atmosphere and the obtained mixture was stirred at 0° C. for 30 min. Methyl iodide (422 mg, 1.49 mmol, 2 equiv) was added and the mixture was stirred for 2 hours at room temperature. The reaction was quenched with saturated aqueous ammonium chloride (20 mL) and the mixture was extracted with ethyl acetate (20 mL×3). The combined organic phases were washed with brine (40 mL×3), dried with sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-80% gradient of ethyl acetate in petroleum ether to afford benzyl N-{2-[4-(1,3-dioxolan-2-yl)-3-methoxyphenyl]ethyl}-N-methylcarbamate (480 mg, 68%) as a light-yellow oil. MS (ESI) calculated for C21H25NO5: 371.17 m/z, found 372.25 [M+H]+.
A solution of benzyl (4-(1,3-dioxolan-2-yl)-3-methoxyphenethyl)(methyl)carbamate (480 mg, 1.29 mmol, 1 equiv) in ethyl acetate (30 mL) was added 10% palladium on carbon (240 mg, 0.2 equiv). The resulting mixture was stirred overnight at room temperature under hydrogen atmosphere. The mixture was diluted with dichloromethane (50 mL) and filtered, rinsing with dichloromethane (3×50 mL). The filtrate was concentrated under reduced pressure to afford {2-[4-(1,3-dioxolan-2-yl)-3-methoxyphenyl]ethyl}(methyl)amine (240 mg, 59%) as a yellow oil. MS (ESI) calculated for C13H19NO3: 237.14 m/z, found 238.20 [M+H]+.
To a solution of 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]benzoic acid (Intermediate 29-2) (200 mg, 0.491 mmol, 1 equiv) and N,N-diisopropylethylamine (127 mg, 0.982 mmol, 2 equiv) in N,N-dimethylformamide (4 mL) were added HATU (187 mg, 0.491 mmol, 1 equiv) and {2-[4-(1,3-dioxolan-2-yl)-3-methoxyphenyl]ethyl}(methyl)amine (116 mg, 0.491 mmol, 1 equiv). The reaction mixture was stirred for 2 h at room temperature. The mixture was purified by reverse-phase column chromatography on C18 silica gel using a 5-70% gradient of acetonitrile in water (+0.05% 2,2,2-trifluoroacetic acid) to afford 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]-N-{2-[4-(1,3-dioxolan-2-yl)-3-methoxyphenyl-]ethyl}-N-methylbenzamide (60 mg, 17%) as a yellow solid. MS (ESI) calculated for C30H21N5O2: 626.26 m/z, found 627.45 [M+H]+.
To a stirred mixture of 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]-N-{2-[4-(1,3-dioxolan-2-yl)-3-methoxyphenyl]ethyl}-N-methylbenzamide (60 mg, 0.096 mmol, 1 equiv) in dichloromethane (5 mL) was added boron tribromide (0.96 mL, 0.96 mmol, 10 equiv) dropwise at −30° C. under nitrogen atmosphere. The resulting mixture was stirred overnight at room temperature under nitrogen atmosphere. The mixture was quenched with saturated aqueous sodium bicarbonate until the pH was ˜6. The resulting mixture was extracted with ethyl acetate (6×30 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by preparative HPLC on a XBridge Shield RP18 OBD Column using a 47-62% gradient of acetonitrile in water (+10 mmol/L ammonium bicarbonate) to afford 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]-N-[2-(4-formyl-3-hydroxyphenyl)ethyl]-N-methylbenzamide (Example 57) (8.4 mg, 15%) as a yellow solid. MS (ESI) calculated for C34H28N6O3: 568.22 m/z, found 569.10 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.13-10.19 (s, 1H), 8.28-8.30 (m, 1H), 8.00-8.07 (m, 4H), 7.43-7.66 (m, 7H), 7.30-7.33 (m, 2H), 6.95-6.60 (m, 1H), 6.46-6.63 (m, 1H), 6.33-6.44 (m, 1H), 3.51-3.71 (m, 1H), 3.40-3.45 (m, 1H), 2.90 (s, 2H), 2.74-2.83 (m, 2H), 2.51-2.73 (m, 1H).
Example 58 was prepared in a manner analogous to Example 29 using Intermediate 11-1 in place of Intermediate 29-2 and 1-(1-benzofuran-6-yl)methanamine in place of Intermediate 29-3. MS (ESI) calculated for C33H26N6O3, 554.21 m/z: found 555.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.16 (s, 1H), 8.26-8.28 (m, 1H), 7.97-8.04 (m, 4H), 7.59-7.61 (m, 1H), 7.40-7.45 (m, 7H), 7.20-7.22 (m, 1H), 6.80-6.88 (m, 2H), 6.38-6.42 (m, 1H), 4.30 (s, 2H), 3.71 (s, 2H).
Example 59 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 59-1 in place of Intermediate 1-4. MS (ESI) calculated for C35H27FN6O3: 598.21 m/z, found 599.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.19 (s, 1H), 8.30-8.45 (m, 1H), 8.00-8.19 (m, 4H), 7.72-7.90 (m, 1H), 7.40-7.60 (m, 6H), 7.30-7.40 (m, 2H), 6.90-7.15 (m, 1H), 6.65-6.90 (m, 1H), 4.20-4.50 (m, 2H), 1.40-1.60 (m, 2H), 1.00-1.30 (m, 2H). 19F NMR (282 MHz, DMSO-d6) δ (ppm): −135.97.
To a stirred solution of 2-{4-bromo-3-fluoro-2-[(4-methoxyphenyl)methoxy]phenyl}-1,3-dioxolane (Intermediate 21-1) (3.00 g, 7.83 mmol, 1 equiv) and tert-butyl[(1-methoxyethenyl)oxy]dimethylsilane (6.00 g, 31.9 mmol, 4 equiv) in N,N-dimethylformamide (12 mL) were added lithium fluoride (450 mg, 17.3 mmol, 2.22 equiv) and bis(tri-tert-butylphosphine)palladium(0) (450 mg, 0.881 mmol, 0.11 equiv) at room temperature under nitrogen atmosphere. The resulting mixture was stirred overnight at 100° C. The reaction was quenched by the addition of water (10 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (3×10 mL). The combined organic layers were washed with brine (3×10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-20% gradient of ethyl acetate in petroleum ether to afford methyl 2-[4-(1,3-dioxolan-2-yl)-2-fluoro-3-[(4-methoxyphenyl)methoxy]phenyl]acetate (2 g, 68%) as a white solid. MS (ESI) calculated for C20H21FO6: 376.13 m/z, found 377.10 [M+H]+.
A mixture of methyl 2-[4-(1,3-dioxolan-2-yl)-2-fluoro-3-[(4-methoxyphenyl) methoxy]phenyl]acetate (1.5 g, 3.985 mmol, 1 equiv), ethenyldiphenylsulfanium triflate (1.8 g, 5.0 mmol, 1.25 equiv) and 1,8-diazabicyclo(5.4.0)undec-7-ene (1.95 g, 12.8 mmol, 3.2 equiv) in dimethyl sulfoxide (75 mL) was stirred overnight at room temperature. The reaction was quenched by the addition of water (100 mL) and the resulting mixture was extracted with ethyl acetate (3×100 mL). The combined organic layers were washed with brine (3×100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-15% gradient of ethyl acetate in petroleum ether to afford methyl 1-[4-(1,3-dioxolan-2-yl)-2-fluoro-3-[(4-methoxyphenyl)methoxy]phenyl]cyclopropane-1-carboxylate (1 g, 62%) as a yellow oil. MS (ESI) calculated for C22H23FO6: 402.15 m/z, found 403.10 [M+H]+.
A mixture of methyl 1-[4-(1,3-dioxolan-2-yl)-2-fluoro-3-[(4-methoxyphenyl)methoxy]phenyl]cyclopropane-1-carboxylate (900 mg, 2.24 mmol, 1 equiv) and lithium hydroxide (180 mg, 7.52 mmol, 3.36 equiv) in tetrahydrofuran (3 mL), water (3 mL) and ethanol (3 mL) was stirred for 2 h at room temperature. The resulting mixture was concentrated under reduced pressure. The mixture was then diluted with water (5 mL) and the pH was brought to 6 with 1N hydrochloric acid. The resulting mixture was extracted with ethyl acetate (3×5 mL). The combined organic layers were washed with brine (3×10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford 1-[4-(1,3-dioxolan-2-yl)-2-fluoro-3-[(4-methoxyphenyl)methoxy]phenyl]cyclopropane-1-carboxylic acid (Intermediate 59-1) (900 mg, crude quant.) as a white solid, which was used in subsequent transformations without further purification. MS (ESI) calculated for C21H21FO6: 388.13 m/z, found 411.15 [M+Na]+.
Example 60 was prepared in a manner analogous to Example 1 (starting from Step 4) using Intermediate 4-1 in place of the amine starting material and Intermediate 60-1 in place of Intermediate 1-4. MS (ESI) calculated for C35H30N6O3, 582.24 m/z: found 583.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.20 (s, 1H), 8.25-8.30 (m, 1H), 8.10-8.15 (m, 1H), 7.95-8.05 (m, 4H), 7.55-7.65 (m, 1H), 7.45-7.50 (m, 2H), 7.35-7.45 (m, 3H), 7.25-7.35 (m, 2H), 7.15-7.25 (m, 1H), 6.90-7.05 (m, 4H), 6.37-6.45 (m, 1H), 4.35-4.37 (m, 2H), 1.50 (m, 6H).
To a solution of 2-(4-bromo-2-((4-methoxybenzyl)oxy)phenyl)-1,3-dioxolane (Intermediate 1-3) (4.15 g, 11.3 mmol, 1 equiv) and tert-butyldimethylsilyl methyl carbonate (8.65 g, 45.4 mmol, 4 equiv) in N,N-dimethylformamide (20 mL) were added bis(tri-tert-butylphosphine)palladium(0) (0.60 g, 1.14 mmol, 0.1 equiv) and lithium fluoride (0.6 g, 23 mmol, 2 equiv). The resulting mixture was stirred at 100° C. under nitrogen atmosphere for 2 h then cooled to 0° C. and quenched with water (100 mL). The resulting mixture was extracted with ethyl acetate (200 mL×3). The combined organic layers were washed with water (100 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a 0-60% gradient of ethyl acetate in petroleum ether to provide methyl 2-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy) phenyl)acetate (3.0 g, 74%) as a light-yellow oil. MS (ESI) calculated for C20H22O6: 358.14 m/z, found 359.10 [M+H]+.
To a cooled (0° C.) solution of methyl 2-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl) methoxy]phenyl]acetate (1.00 g, 2.79 mmol, 1 equiv) in tetrahydrofuran (5 mL) were added dropwise a solution of potassium tert-butoxide (9 mL, 9.00 mmol, 3.2 equiv, 1M in tetrahydrofuran) and methyl iodide (1.66 g, 11.7 mmol, 4.2 equiv) with stirring. The resulting mixture was stirred at room temperature for 2 h. The reaction was quenched by addition of 1M aqueous potassium bisulfate, and the mixture was extracted with ethyl acetate. The organic extract was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford methyl 2-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl) methoxy]phenyl]-2-methylpropanoate (680 mg, 63%) as a yellow solid, which was used without purification in the next step. MS (ESI) calculated for C22H26O6: 386.17 m/z, found 387.15 [M+H]+.
To a stirred solution of methyl 2-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl) methoxy]phenyl]-2-methylpropanoate (600 mg, 1.55 mmol, 1 equiv) in tetrahydrofuran (10 mL) and methanol (10 mL) was added a solution of lithium hydroxide (74 mg, 3.1 mmol, 2 equiv) in water (10 mL). The resulting mixture was stirred for 2 h at room temperature. The mixture was acidified to pH 5-6 with hydrochloric acid (1M). The resulting mixture was extracted with ethyl acetate (3×50 mL) and the combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford 2-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]-2-methylpropanoic acid (Intermediate 60-1) (500 mg, 86%) as a brown oil. MS (ESI) calculated for C21H24O6: 372.16 m/z, found 373.15 [M+H]+.
Example 61 was prepared in a manner analogous to Example 1 using pyridin-4-ylboronic acid in place of 4-(morpholin-4-yl)phenylboronic acid. MS (ESI) calculated for C32H25N7O3: 555.20 m/z, found 556.00 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.22 (s, 1H), 8.78-8.95 (m, 2H), 8.45-8.52 (m, 1H), 8.30-8.45 (m, 3H), 8.02-8.12 (m, 1H), 7.68-7.80 (m, 1H), 7.58-7.68 (m, 1H), 7.48-7.58 (m, 2H), 7.38-7.48 (m, 2H), 6.82-7.02 (m, 2H), 6.65-6.82 (m, 1H), 4.40 (s, 2H), 3.58 (s, 2H).
Example 62 was prepared in a manner analogous to Example 1 (starting from Step 2) using Intermediate 62-2 in place of the Boc-protected starting material. MS (ESI) calculated for C32H25N7O3: 555.20 m/z, found 556.00 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.22 (s, 1H), 8.68-8.78 (m, 1H), 8.48-8.62 (m, 1H), 8.35-8.48 (m, 1H), 8.15-8.28 (m, 1H), 8.05-8.15 (m, 1H), 7.88-8.05 (m, 1H), 7.75-7.88 (m, 1H), 7.58-7.68 (m, 1H), 7.50-7.58 (m, 2H), 7.28-7.50 (m, 3H), 6.80-7.00 (m, 3H), 4.40 (s, 2H), 3.58 (s, 2H).
To a solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-chloroimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 1-1) (500 mg, 1.11 mmol, 1 equiv) and 2-(tributylstannyl)pyridine (612 mg, 1.66 mmol, 1.5 equiv) in toluene (5 mL) were added lithium chloride (188 mg, 4.44 mmol, 4 equiv) and bis(triphenylphosphine)palladium(II) dichloride (78 mg, 0.11 mmol, 0.1 equiv). The resulting mixture was stirred overnight at 100° C. under nitrogen atmosphere then cooled to room temperature and concentrated under reduced pressure. The residue was purified by reverse-phase flash column chromatography on C18 silica gel using a 10-50% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) to afford tert-butyl N-({4-[2-(2-aminopyridin-3-yl)-5-(pyridin-2-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (Intermediate 62-1) (300 mg, 53%) as a brown solid. MS (ESI) calculated for C28H27N7O2: 493.22 m/z, found 494.25 [M+H]+.
Example 63 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 13-2 in place of Intermediate 1-4. MS (ESI) calculated for C33H25ClN6O3: 588.17 m/z, found 589.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.08 (s, 1H), 8.74 (s, 1H), 8.26-8.28 (m, 1H), 7.98-8.02 (m, 4H), 7.67-7.69 (m, 1H), 7.40-7.49 (m, 7H), 7.11-7.23 (m, 2H), 6.41-6.44 (m, 1H), 4.41-4.43 (m, 2H), 3.79-3.80 (m, 2H).
Example 64 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 64-1 in place of the amine starting material. MS (ESI) calculated for C34H28N6O3: 568.22 m/z, found 569.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm) 10.23 (s, 1H), 8.30-8.41 (m, 1H), 8.01-8.11 (m, 4H), 7.63-7.70 (m, 1H), 7.60-7.70 (m, 1H), 7.40-7.62 (m, 7H), 6.91-7.01 (m, 2H), 6.75-6.90 (m, 1H), 4.61-4.80 (m, 2H), 3.81-3.98 (m, 2H), 3.10-2.85 (m, 3H).
Intermediate 64-1 was prepared in a manner analogous to Intermediate 11-1 using tert-butyl N-[(4-aminophenyl)methyl]-N-methylcarbamate in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C25H22N6: 406.19 m/z, found 407.19 [M+H]+.
Example 65 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 65-1 in place of Intermediate 1-4. MS (ESI) calculated for C33H25FN6O3: 572.20 m/z, found 573.10 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.25 (s, 1H), 9.22-9.26 (m, 1H), 8.32-8.35 (m, 1H), 8.01-8.06 (m, 4H), 7.70-7.75 (m, 2H), 7.39-7.52 (m, 7H), 7.07-7.09 (m, 1H), 6.80-7.06 (m, 1H), 6.75-6.78 (m, 1H), 5.96-6.12 (m, 1H), 4.44-4.46 (m, 2H). 19F NMR (300 MHz, DMSO-d6) δ (ppm): −181.34.
A mixture of 2-{4-bromo-2-[(4-methoxyphenyl)methoxy]phenyl}-1,3-dioxolane (Intermediate 1-3) (1.00 g, 2.74 mmol, 1 equiv), ethyl 2-chloro-2-fluoroacetate (0.77 g, 5.5 mmol, 2 equiv), nickel(II) iodide (0.086 g, 0.28 mmol, 0.10 equiv), 5,5′-dimethyl-2,2′-bipyridine (0.030 g, 0.16 mmol, 0.06 equiv), BINAP (0.086 g, 0.14 mmol, 0.05 equiv), magnesium(II) chloride (0.39 g, 4.1 mmol, 1.5 equiv) and zinc (0.54 g, 8.2 mmol, 3 equiv) in N,N-dimethylacetamide (20 mL) and tetrahydrofuran (8 mL) was stirred overnight at 80° C. The reaction was quenched by the addition of water (30 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (50 mL×3). The combined organic layers were washed with water (50 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-25% gradient of ethyl acetate in petroleum ether to afford ethyl 2-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]-2-fluoroacetate (460 mg, 40%) as a yellow oil. 1H-NMR (400 MHz, DMSO-d6) δ (ppm): 7.50-7.58 (m, 1H), 7.35-7.48 (m, 2H), 7.12-7.21 (m, 1H), 6.99-7.08 (m, 1H), 6.90-6.98 (m, 2H), 6.01-6.21 (m, 1H), 5.69-5.79 (m, 1H), 5.02-5.19 (m, 2H), 4.26-4.29 (m, 2H), 4.01-4.09 (m, 2H), 3.89-3.99 (m, 2H), 3.70-3.75 (m, 3H), 1.12-1.23 (m, 3H). 19F-NMR (300 MHz, DMSO-d6) δ (ppm): −176.55.
A solution of ethyl 2-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]-2-fluoroacetate (700 mg, 1.79 mmol, 1 equiv) and lithium hydroxide (86 mg, 3.6 mmol, 2 equiv) in ethanol (10 mL), water (10 mL) and tetrahydrofuran (10 mL) was stirred for 2 h at room temperature. The mixture was partially concentrated in vacuo and diluted with water (30 mL). The mixture was acidified to pH 5-6 with hydrochloric acid (0.05 M) and extracted with ethyl acetate (50 mL×3). The combined organic layers were washed with water (50 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford [4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl](fluoro)acetic acid (Intermediate 65-1) (450 mg, 69%) as a yellow oil, which was used in subsequent transformations without further purification. MS (ESI) calculated for C19H19FO6: 362.12 m/z, found 361.00 [M−H]+.
Example 66 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 16-2 in place of the amine starting material. MS (ESI) calculated for C30H26N6O3: 518.21 m/z, found 519.25 [M+H]+. 1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.18 (s, 1H), 8.08-8.11 (m, 1H), 8.02-8.04 (m, 1H), 7.65-7.67 (m, 1H), 7.66-7.69 (m, 1H), 7.61-7.64 (m, 4H), 7.31-7.41 (m, 1H), 6.93-6.96 (m, 1H), 6.90-6.92 (m, 1H), 6.74-6.79 (m, 1H), 4.37 (s, 2H), 3.65 (s, 2H), 2.18-2.21 (m, 1H), 0.94-1.00 (m, 2H), 0.82-0.83 (m, 2H).
Example 67 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 67-1 in place of the amine starting material and Intermediate 59-1 in place of Intermediate 1-4. MS (ESI) calculated for C34H26FN7O3: 599.21 m/z, found 600.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.22 (s, 1H), 9.20-9.30 (m, 1H), 8.65-8.85 (m, 2H), 8.33-8.55 (m, 1H), 8.19-8.30 (m, 1H), 8.00-8.19 (m, 1H), 7.73-7.94 (m, 2H), 7.45-7.64 (m, 3H), 7.25-7.45 (m, 2H), 7.00-7.10 (m, 1H), 6.70-6.95 (m, 1H), 4.25-4.45 (m, 2H), 1.40-1.60 (m, 2H), 1.00-1.30 (m, 2H). 19F NMR (282 MHz, DMSO-d6) δ (ppm): −136.09.
Intermediate 67-1 was prepared in a manner analogous to Intermediate 12-1 using pyridin-3-ylboronic acid in place of (3-morpholinophenyl)boronic acid. MS (ESI) calculated for C23H19N7: 393.17 m/z, found 394.10 [M+H]+.
Example 68 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 67-1 in place of the amine starting material. MS (ESI) calculated for C32H25N7O3: 555.20 m/z, found 556.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.20 (s, 1H), 9.28 (s, 1H), 8.62-8.80 (m, 1H), 8.45-8.60 (m, 1H), 8.35-8.45 (m, 1H), 8.10-8.20 (m, 1H), 8.00-8.10 (m, 1H), 7.60-7.75 (m, 3H), 7.50-7.60 (m, 2H), 7.40-7.50 (m, 2H), 6.80-7.05 (m, 2H), 6.70-6.80 (m, 1H), 4.35-4.40 (m, 2H), 3.50-3.75 (m, 2H).
Example 69 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 69-1 in place of Intermediate 1-4. MS (ESI) calculated for C35H28N6O3: 580.22 m/z, found 581.23 [M+H]+. 1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.23 (s, 1H), 8.34-8.32 (m, 1H), 8.01-8.07 (m, 4H), 7.65-7.73 (m, 2H), 7.41-7.63 (m, 5H), 7.33-7.40 (m, 2H), 6.97-7.00 (m, 2H), 6.75-6.79 (m, 1H), 4.32 (s, 2H), 1.39-1.42 (m, 2H), 1.04-1.07 (m, 2H).
Intermediate 69-1 was prepared in a manner analogous to Intermediate 59-1 using Intermediate 1-3 in place of Intermediate 21-1.MS (ESI) calculated for C21H22O6: 370.14 m/z, found 371.10 [M+H]+.
Example 70 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 67-1 in place of the amine staring material and Intermediate 69-1 in place of Intermediate 1-4. MS (ESI) calculated for C34H27N7O3: 581.22 m/z, found 582.22 [M+H]+. 1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.19 (s, 1H), 9.20-9.26 (m, 1H), 8.63-8.70 (m, 2H), 8.40-8.42 (m, 1H), 8.14-8.17 (m, 1H), 8.04-8.06 (m, 1H), 7.74-7.78 (m, 2H), 7.63-7.65 (m, 1H), 7.47-7.49 (m, 2H), 7.32-7.34 (m, 2H), 6.96-6.98 (m, 2H), 6.77-6.81 (m, 1H), 4.31 (s, 2H), 1.38-1.40 (m, 2H) 1.06-1.10 (m, 2H).
Example 71 was prepared in a manner analogous to Example 12 using Intermediate 67-1 in place of Intermediate 12-1, 3-formyl-4-hydroxybenzoic acid in place of 3-fluoro-5-formyl-4-hydroxybenzoic acid and HOBT in place of HATU. MS (ESI) calculated for C3H23N7O2: 541.19 m/z, found 542.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.32 (s, 1H), 9.25-9.32 (m, 1H), 8.65-8.72 (m, 1H), 8.51-8.65 (m, 1H), 8.38-8.51 (m, 1H), 8.27-8.38 (m, 1H), 8.16-8.27 (m, 1H), 8.01-8.16 (m, 2H), 7.62-7.87 (m, 2H), 7.41-7.62 (m, 4H), 6.97-7.32 (m, 1H), 6.67-6.89 (m, 1H), 4.45-4.78 (m, 2H).
Example 72 was prepared in a manner analogous to Example 19 using Intermediate 36-1 in place of Intermediate 19-2. MS (ESI) calculated for C33H28N6O2: 540.23 m/z, found 541.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.30 (s, 1H), 8.30-8.32 (m, 1H), 8.00-8.04 (m, 4H), 7.74-7.76 (m, 1H), 7.11-7.55 (m, 8H), 6.66-7.09 (m, 2H), 6.63-6.64 (m, 1H), 4.21 (s, 2H), 3.27-3.31 (m, 2H), 3.03-3.07 (m, 2H).
Example 73 was prepared in a manner analogous to Example 12 using Intermediate 4-1 in place of Intermediate 12-1, 3-formyl-4-hydroxybenzoic acid in place of 3-fluoro-5-formyl-4-hydroxybenzoic acid and HOBT in place of HATU. MS (ESI) calculated for C32H24N6O3: 540.19 m/z, found 541.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.30 (s, 1H), 8.15-8.43 (m, 2H), 7.90-8.19 (m, 5H), 8.62-8.85 (m, 1H), 7.30-7.60 (m, 7H), 7.05-7.25 (m, 1H), 6.60-6.90 (m, 1H), 4.45-4.80 (m, 2H).
Example 74 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 74-1 in place of the amine starting material and Intermediate 65-1 in place of Intermediate 1-4. MS (ESI) calculated for C27H21FN6O3: 496.17 m/z, found 497.15 [M+H]+. 1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.16 (s, 1H), 8.37-8.38 (m, 1H), 8.23-8.25 (m, 1H), 7.95-7.97 (m, 1H), 7.70-7.73 (m, 2H), 7.43-7.47 (m, 1H), 7.35-7.36 (m, 4H), 7.04-7.07 (m, 2H), 6.71-6.75 (m, 1H), 5.90-6.01 (m, 1H), 4.39-4.40 (m, 2H). 19F NMR (300 MHz, DMSO-d6) δ (ppm): 181.28.
To a solution of tert-butyl (4-(2-(2-aminopyridin-3-yl)-5-chloro-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)carbamate (Intermediate 1-1) (100 mg, 0.285 mmol, 1 equiv) in methanol (10 mL) was added palladium on carbon (10%, 50 mg, 0.15 equiv) and sodium bicarbonate (96 mg, 1.1 mmol, 4 equiv) under nitrogen atmosphere. The mixture stirred under hydrogen atmosphere at room temperature for 1 h. The resulting mixture was filtered through celite and concentrated in vacuo to afford 3-{3-[4-(aminomethyl)phenyl]imidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (80 mg, 89%) as a yellow solid, which was used in the next step without further purification. MS (ESI) calculated for C18H16N6: 316.16 m/z, found 317.15 [M+H]+.
A solution of tert-butyl N-({4-[2-(2-aminopyridin-3-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}methyl)carbamate (300 mg, 0.720 mmol, 1 equiv) in 4N hydrochloric acid in 1,4-dioxane (5 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated in vacuo and purified by reverse-phase flash column chromatography on C18 silica gel using a 20-95% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) with a 10 minute hold at 70% acetonitrile to afford 3-{3-[4-(aminomethyl)phenyl]imidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 74-1) as a yellow solid (200 mg, 88%). MS (ESI) calculated for C18H16N6: 316.14 m/z, found 317.05 [M+H]+.
Example 75 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material, Intermediate 75-1 in place of Intermediate 1-4 and hydrogen bromide (33% in acetic acid) overnight instead of 2,2,2-trifluoroacetic acid/methanesulfonic acid for 1 h. MS (ESI) calculated for C32H25N7O3: 555.20 m/z, found 556.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.09 (s, 1H), 8.20-8.40 (m, 1H), 7.85-8.05 (m, 5H), 7.55-7.75 (m, 1H), 7.35-7.52 (m, 7H), 6.70-6.80 (m, 1H), 6.30-6.40 (m, 1H), 4.41 (s, 2H), 3.57 (s, 2H).
To a cooled (−30° C.) mixture of methyl 6-bromo-2-methoxypyridine-3-carboxylate (4.00 g, 16.3 mmol, 1 equiv) in tetrahydrofuran (94 mL) was added di-iso-butyl aluminum hydride (1M in toluene, 49 mL, 49 mmol, 3 equiv) dropwise with stirring. The resulting mixture was stirred for an additional 2 hours at room temperature and the reaction was quenched by the addition of saturated aqueous potassium sodium tartrate (200 mL) and methanol (100 mL) at 0° C. The resulting mixture was extracted with ethyl acetate (3×100 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford (6-bromo-2-methoxypyridin-3-yl) methanol (3.93 g, crude quant.) as a pale-yellow solid, which was used without further purification in the next step. MS (ESI) calculated for C7H8BrNO2: 216.97 m/z, found 218.05, 220.05 [M+H, M+2+H]+.
A suspension of (6-bromo-2-methoxypyridin-3-yl) methanol (54 mg, 0.25 mmol, 1 equiv) and manganese (IV) oxide (322 mg, 3.71 mmol, 15 equiv) in 1,2-dichloroethane (2 mL) was stirred overnight at room temperature. The resulting mixture was filtered, rinsing with ethyl acetate (3×50 mL). The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography using a 0-20% gradient of ethyl acetate in petroleum ether to afford 6-bromo-2-methoxypyridine-3-carbaldehyde (2.84 g, 74%) as a pale-yellow solid. MS (ESI) calculated for C7H6BrNO2: 214.96 m/z, found 216.05, 218.10 [M+H, M+2+H]+.
A solution of 6-bromo-2-methoxypyridine-3-carbaldehyde (2.84 g, 13.1 mmol, 1 equiv) in toluene (150 mL) was treated with ethylene glycol (2.45 g, 39.4 mmol, 3 equiv) and p-toluenesulfonic acid (234 mg, 1.36 mmol, 0.10 equiv). The resulting mixture was stirred for 10 min at room temperature then overnight at 90° C. The mixture was allowed to cool to room temperature and was quenched with water (200 mL). The resulting mixture was extracted with ethyl acetate (3×100 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography, using a 0-15% gradient of ethyl acetate in petroleum ether to afford 6-bromo-3-(1,3-dioxolan-2-yl)-2-methoxypyridine (3.2 g, 94%) as a pale-yellow oil. MS (ESI) calculated for C9H10BrNO3: 258.98 m/z, found 259.90, 261.90 [M+H, M+2+H]+.
A solution of 6-bromo-3-(1,3-dioxolan-2-yl)-2-methoxypyridine (815 mg, 3.13 mmol, 1 equiv), bis(tri-tert-butylphosphine)palladium(0) (320 mg, 0.627 mmol, 0.2 equiv), lithium fluoride (325 mg, 12.5 mmol, 4 equiv) and tert-butyl[(1-methoxyethenyl)oxy]dimethylsilane (4.721 mg, 25.07 mmol, 8 equiv) in N,N-dimethylformamide (8 mL) was stirred for 3 hours at 100° C. under nitrogen atmosphere. The reaction was quenched with water at room temperature. The resulting mixture was extracted with dichloromethane (3×50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-45% gradient of 10:1 dichloromethane/methanol in petroleum ether to afford methyl 2-[5-(1,3-dioxolan-2-yl)-6-methoxypyridin-2-yl]acetate (900 mg, quant.) as a reddish-brown oil. MS (ESI) calculated for C12H15NO5: 253.10 m/z, found 254.15 [M+H]+.
To a cooled (0° C.) solution of methyl 2-[5-(1,3-dioxolan-2-yl)-6-methoxypyridin-2-yl]acetate (1.29 g, 5.09 mmol, 1 equiv) in tetrahydrofuran (10 mL) and ethanol (10 mL) was added a solution of lithium hydroxide (0.24 g, 10 mmol, 2 equiv) in water (5 mL) dropwise. The resulting mixture was stirred for 1 h at room temperature. The mixture was partially concentrated and brought to a pH of 6 with 0.5 M hydrochloric acid. The resulting mixture was concentrated under reduced pressure and suspended in dichloromethane/methanol (20 mL). The mixture was filtered, rinsing with dichloromethane/methanol (3×20 mL). The filtrate was concentrated under reduced pressure to afford [5-(1,3-dioxolan-2-yl)-6-methoxypyridin-2-yl]acetic acid (Intermediate 75-1) (550 mg, 38%) as brown/yellow solid, which was used in subsequent transformations without further purification. MS (ESI) calculated for C11H13NO5: 239.08 m/z, found 240.05 [M+H]+.
Example 76 was prepared in a manner analogous to Example 19 using Intermediate 4-1 in place of Intermediate 19-2, Intermediate 76-1 in place of Intermediate 19-3 and dichloromethane/2,2,2-trifluoroacetic acid (15:1) in place of 2,2,2-trifluoroacetic acid/methanesulfonic acid. MS (ESI) calculated for C34H30N6O3: 570.24 m/z, found 571.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ ppm: 10.07 (m, 1H), 8.23-8.33 (m, 1H), 7.85-8.11 (m, 4H), 7.39-7.52 (m, 8H), 7.09-7.21 (m, 1H), 6.86-7.06 (m, 1H), 6.23-6.38 (m, 1H), 3.69-3.85 (m, 5H), 2.76-2.88 (m, 4H).
To a solution of 3-bromo-2-methoxyphenol (2.00 g, 9.85 mmol, 1 equiv) in acetonitrile (15 mL) were added paraformaldehyde (4.44 g, 49.3 mmol, 5 equiv), magnesium(II) chloride (1.41 g, 14.8 mmol, 1.5 equiv) and triethylamine (2.49 g, 24.6 mmol, 2.5 equiv). The obtained suspension was stirred at room temperature for 10 min then at 80° C. for 2 h. The resulting mixture was cooled to 0° C. and quenched by the addition of hydrochloric acid (2 M). The resulting mixture was extracted with dichloromethane (30 mL×3). The combined organic layers were washed with water (20 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-7% gradient of ethyl acetate in petroleum ether to provide 4-bromo-2-hydroxy-3-methoxybenzaldehyde (1.28 g, 56%) as a light-yellow oil. MS (ESI) calculated for C8H7BrO3: 229.96 m/z, found 230.90 [M+H]+.
To a colorless solution of 4-bromo-2-hydroxy-3-methoxybenzaldehyde (1.28 g, 5.54 mmol, 1 equiv) in toluene (13 mL) were added ethylene glycol (1.72 g, 27.7 mmol, 5 equiv), triethyl orthoformate (2.46 g, 16.6 mmol, 3 equiv) and p-toluenesulfonic acid (0.10 g, 0.55 mmol, 0.1 equiv). The obtained solution was stirred at room temperature for 10 min then overnight at 90° C. The resulting solution was cooled to 0° C. and quenched by the addition of water (5 mL). The resulting mixture was extracted with ethyl acetate (10 mL×3). The combined organic layers were washed with brine (10 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-7% gradient of ethyl acetate in petroleum ether to provide 3-bromo-6-(1,3-dioxolan-2-yl)-2-methoxyphenol (0.58 g, 38%) as a white solid. MS (ESI) calculated for C10H11BrO4: 273.98 m/z, found 274.90 [M+H]+.
To a solution of 3-bromo-6-(1,3-dioxolan-2-yl)-2-methoxyphenol (0.58 g, 2.11 mmol, 1 equiv) and imidazole (0.29 g, 4.2 mmol, 2 equiv) in N,N-dimethylformamide (15 mL) was added tert-butyldimethylsilyl chloride (0.44 g, 3.0 mmol, 1.4 equiv). The resulting mixture was stirred overnight at room temperature then cooled to 0° C. and quenched by the addition of water (20 mL). The resulting mixture was extracted with ethyl acetate (30 mL×3). The combined organic layers were washed with brine (30 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-7% gradient of ethyl acetate in petroleum ether to provide 3-bromo-6-(1,3-dioxolan-2-yl)-2-methoxyphenoxy(tert-butyl)dimethylsilane (0.57 g, 69%) as a white solid. MS (ESI) calculated for C1H25BrO4Si: 388.07 m/z, found 389.10 [M+H]+.
To a solution of 3-bromo-6-(1,3-dioxolan-2-yl)-2-methoxyphenoxy(tert-butyl)dimethylsilane (0.64 g, 1.6 mmol, 1 equiv) and tributyl(prop-2-en-1-yl)stannane (1.09 g, 3.29 mmol, 2 equiv) in N,N-dimethylformamide (6 mL) was added bis(triphenylphosphine )palladium(II) dichloride (0.12 g, 0.16 mmol, 0.1 equiv). After stirring for 1 h at 80° C. under a nitrogen atmosphere, the mixture was cooled to 0° C. and quenched by the addition of saturated aqueous ammonium chloride (10 mL). The resulting mixture was extracted with ethyl acetate (20 mL×3). The combined organic layers were washed with water (10 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-30% gradient of ethyl acetate in petroleum ether to provide tert-butyl(6-(1,3-dioxolan-2-yl)-2-methoxy-3-(prop-2-en-1-yl)phenoxy)dimethylsilane (0.57 g, 99%) as a white solid. MS (ESI) calculated for C19H30O4Si: 350.19 m/z, found 351.20 [M+H]+.
To a solution of tert-butyl(6-(1,3-dioxolan-2-yl)-2-methoxy-3-(prop-2-en-1-yl) phenoxy)dimethylsilane (0.54 g, 1.5 mmol, 1 equiv) in tetrahydrofuran (10 mL) and water (10 mL) were added osmium tetroxide (0.54 mL, 10 mmol, 6.8 equiv) and sodium periodate (0.99 g, 4.6 mmol, 3 equiv). After stirring for 30 min at room temperature, the reaction was quenched by addition of water. The resulting mixture was extracted with ethyl acetate (60 mL×3). The combined organic layers were washed with water (50 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-25% gradient of ethyl acetate in petroleum ether to provide 2-{3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)-2-methoxyphenyl}acetaldehyde (Intermediate 76-1) (0.12 g, 22%) as a light-yellow solid. MS (ESI) calculated for C18H28O5Si: 352.17 m/z, found 353.20 [M+H]+.
Example 77 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 77-1 in place of Intermediate 1-4. Mass (ESI) calculated for C32H25N7O3: 555.20, found 556.10 [M+H]+. 1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.04 (s, 1H), 8.25-8.27 (m, 1H), 7.97-8.02 (m, 5H), 7.34-7.43 (m, 8H), 7.20-7.22 (m, 1H), 6.30-6.42 (m, 1H), 4.39-4.40 (s, 2H), 3.50 (s, 2H).
Intermediate 77-1 was prepared in a manner analogous to Intermediate 56-1 using benzyl bromide in place of p-methoxybenzyl chloride/potassium iodide and omitting Step 6 (cyclopropanation). MS (ESI) calculated for C17H17NO5: 315.11 m/z, found 316.00 [M+H]+.
Example 78 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 74-1 in place of the amine starting material and Intermediate 78-1 in place of Intermediate 1-4. MS (ESI) calculated for C27H20F2N6O3: 514.16 m/z, found 515.10 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.35 (s, 1H), 9.68-9.78 (m, 1H), 8.16-8.50 (m, 2H), 7.90-8.10 (m, 1H), 7.70-7.75 (m, 1H), 7.33-7.50 (m, 5H), 7.20-7.28 (m, 2H), 7.15-7.20 (m, 1H), 6.95-6.90 (m, 2H), 6.45-6.35 (m, 1H), 4.45-4.38 (m, 2H). 19F NMR (300 MHz, DMSO-d6) δ (ppm): −102.77.
A mixture of 2-{4-bromo-2-[(4-methoxyphenyl)methoxy]phenyl}-1,3-dioxolane (Intermediate 1-3) (500 mg, 1.37 mmol, 1 equiv), ethyl 2-bromo-2,2-difluoroacetate (834 mg, 4.11 mmol, 3 equiv), XantPhos (119 mg, 0.205 mmol, 0.15 equiv), zinc (269 mg, 4.11 mmol, 3 equiv), tetra-n-butylammonium bromide (662 mg, 2.05 mmol, 1.5 equiv) and palladium(71-cinnamyl) chloride dimer (35 mg, 0.068 mmol, 0.05 equiv) in tetrahydrofuran (20 mL) was stirred overnight at 60° C. under nitrogen atmosphere. The mixture was cooled to room temperature and quenched with water (20 mL). The resulting mixture was extracted with ethyl acetate (20 mL×3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-100% gradient of ethyl acetate in petroleum ether to afford ethyl 2-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]-2,2-difluoroacetate (100 mg, 18%) as a colorless oil. MS (ESI) calculated for C21H22F2O6: 408.14 m/z, found 409.20 [M+H]+.
To a solution of ethyl 2-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]-2,2-difluoroacetate (180 mg 44.1 mmol, 1 equiv) in tetrahydrofuran (10 mL) and ethanol (0.9 mL) was added a solution of lithium hydroxide (88 mg, 88 mmol, 2 equiv) in water (2.5 mL) and the resulting mixture was stirred overnight at room temperature. The reaction was quenched with water (30 mL) and adjusted to pH 6-7 with 1M hydrochloric acid. The mixture was extracted with ethyl acetate (3×40 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford 2-(4-(1,3-dioxolan-2-yl)-3-((4-methoxybenzyl)oxy)phenyl)-2,2-difluoroacetic acid (Intermediate 78-1) (80 mg, 48%) as a colorless oil, which was used without further purification in subsequent transformations. MS (ESI) calculated for C19H18F2O6: 380.11 m/z, found 379.10 [M−H]−.
Example 79 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 67-1 in place of the amine starting material and Intermediate 21-2 in place of Intermediate 1-4. MS (ESI) calculated for C32H24FN703: 573.19 m/z, found 574.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.20 (s, 1H), 9.20-9.22 (m, 1H), 8.58-8.60 (m, 1H), 8.30-8.38 (m, 2H), 8.00-8.08 (m, 2H), 7.40-7.52 (m, 6H), 7.22-7.25 (m, 1H), 6.87-6.92 (m, 1H), 6.41-6.45 (m, 1H), 4.41-4.42 (m, 2H), 3.59-3.65 (m, 2H). 19F-NMR (400 MHz, DMSO-d6) δ (ppm): −139.31.
Example 80 was prepared in a manner analogous to Example 19 using Intermediate 74-1 in place of Intermediate 19-2, Intermediate 80-2 in place of Intermediate 19-3 and dichloromethane/2,2,2-trifluoroacetic acid (10:1) in place of 2,2,2-trifluoroacetic acid/methanesulfonic acid. MS (ESI) calculated for C28H24F2N6O: 498.20 m/z, found 499.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.14 (s, 1H), 8.31-8.33 (m, 1H), 8.18-8.21 (m, 1H), 7.96-8.00 (m, 2H), 7.58-7.76 (m, 3H), 7.38-7.46 (m, 2H), 7.35-7.36 (m, 1H), 7.28-7.32 (m, 2H), 6.97-7.19 (m, 1H), 6.32-6.36 (m, 1H), 3.79 (s, 2H), 2.92-2.94 (m, 2H), 2.78-2.89 (m, 2H).
Intermediate 80-2 was prepared in a manner analogous to Intermediate 16-4 using Intermediate 80-1 in place of Intermediate 16-3. MS (ESI) calculated for C12H12F2O3: 242.08 m/z, found 243.10 [M+H]*.
To a solution of methyl 4-bromo-2-formylbenzoate (1.80 g, 7.41 mmol, 1 equiv) in dichloromethane were added Deoxo-Fluor (4.92 g, 22.2 mmol, 3 equiv) and methanol (0.06 mL) and the resulting mixture was stirred at 60° C. overnight. The mixture was cooled room temperature and brough to pH ˜8 with saturated aqueous sodium bicarbonate. The resulting mixture was extracted with dichloromethane (100 mL×3). The combined organic layers were washed with water (100 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-16% gradient of ethyl acetate in petroleum ether to afford methyl 4-bromo-2-(difluoromethyl)benzoate (1.6 g, 82%) as a white solid. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 7.92-7.95 (m, 3H), 7.33-7.69 (m, 1H), 3.87 (s, 3H).
To a cooled (0° C.) solution of methyl 4-bromo-2-(difluoromethyl)benzoate (1.5 g, 5.7 mmol, 1 equiv) in tetrahydrofuran (100 mL) was added lithium aluminum hydride (1M in tetrahydrofuran, 2.5 mL, 2.5 mmol, 0.5 equiv) under nitrogen atmosphere and the obtained solution was stirred at room temperature for 2 h. The reaction was quenched by the addition of water. The resulting mixture was extracted with ethyl acetate (150 mL×3). The combined organic layers were washed with water (150 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-30% gradient of ethyl acetate in petroleum ether to provide [4-bromo-2-(difluoromethyl)phenyl]methanol (1.3 g, 97%) as a colorless oil. MS (ESI) calculated for C8H7BrF2O: 235.96 m/z, found 234.90 [M−H]−.
To a solution of [4-bromo-2-(difluoromethyl)phenyl]methanol (1.3 g, 5.5 mmol, 1 equiv) in 1,2-dichloroethane (50 mL) was added manganese (IV) oxide (10.0 g, 115 mmol, 21 equiv). After stirring for 2 h at 50° C., the mixture was concentrated under reduced pressure. Water was added and the resulting mixture was extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-30% gradient of ethyl acetate in petroleum ether to provide 4-bromo-2-(difluoromethyl)benzaldehyde (1.0 g, 78%) as a light-yellow solid. MS (ESI) calculated for C8H5BrF2O: 233.95 m/z, found 232 90 [M−H]−.
To a solution of 4-bromo-2-(difluoromethyl)benzaldehyde (60 mg, 0.26 mmol, 1 equiv) in toluene (4 mL) were added ethylene glycol (79 mg, 1.3 mmol, 5 equiv), triethyl orthoformate (113 mg, 0.765 mmol, 3 equiv) and p-toluenesulfonic acid (4.4 mg, 0.026 mmol, 0.1 equiv) and the resulting was stirred at room temperature for 10 min then at 90° C. overnight. The resulting solution was cooled to 0° C. and quenched by the addition of water (5 mL). The mixture was extracted with ethyl acetate (10 mL×3). The combined organic layers were washed with water (10 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-7% gradient of ethyl acetate in petroleum ether to provide 2-[4-bromo-2-(difluoromethyl)phenyl]-1,3-dioxolane (Intermediate 80-1) (40 mg, 56%)) as a white solid. MS (ESI) calculated for C10H9BrF2O2: 277.98 m/z, found 278.90 [M+H]+.
To a stirred solution of tert-butyl(2-(1,3-dioxolan-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)dimethylsilane (Intermediate 81-2) (300 mg, 0.738 mmol) and 3-[3-(4-{[(4-bromopyridin-2-yl)amino]methyl}phenyl)-5-phenylimidazo[4,5-b]pyridin-2-yl]pyridin-2-amine (Intermediate 81-1) (300 mg, 0.547 mmol) in 1,4-dioxane (7.5 mL) and water (2.5 mL) were added [1,1′-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (40 mg, 0.061 mmol) and tribasic potassium phosphate (240 mg, 1.13 mmol) under nitrogen atmosphere. The resulting mixture was stirred for 3 h at room temperature. The reaction was quenched by the addition of water (10 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (3×10 mL). The combined organic layers were washed with brine (3×10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using a 5-60% gradient of acetonitrile in water (+0.05% 2,2,2-trifluoroacetic acid) to afford 3-[3-(4-{[(4-{4-[(tert-butyldimethylsilyl)oxy]-3-(1,3-dioxolan-2-yl)phenyl}pyridin-2-yl)amino]methyl}phenyl)-5-phenylimidazo[4,5-b]pyridin-2-yl]pyridin-2-amine (280 mg, 51%) as a yellow solid. MS (ESI) calculated for C44H45N7O3Si: 747.34 m/z, found 748.35 [M+H]+.
A solution of 3-[3-(4-{[(4-{4-[(tert-butyldimethylsilyl)oxy]-3-(1,3-dioxolan-2-yl)phenyl}pyridin-2-yl)amino]methyl}phenyl)-5-phenylimidazo[4,5-b]pyridin-2-yl]pyridin-2-amine (90 mg, 0.12 mmol), potassium fluoride (20 mg, 0.34 mmol) and hydrogen bromide (0.2 mL, 30% in water) in N,N-dimethylformamide (1 mL) was stirred for 3 h at room temperature. The mixture was purified by preparative HPLC on a XBridge Shield RP18 OBD Column using a 48-64% gradient of acetonitrile in water (+10 mmol/L ammonium bicarbonate) to provide 5-{2-[({4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)amino]pyridin-4-yl}-2-hydroxybenzaldehyde (Example 81) (6 mg, 8%) as a yellow solid. MS (ESI) calculated for C36H27N7O2: 589.22 m/z, found 590.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.29 (s, 1H), 8.20-8.30 (m, 1H), 7.80-8.15 (m, 7H), 7.50-7.62 (m, 2H), 7.40-7.50 (m, 5H), 7.15-7.29 (m, 1H), 7.00-7.18 (m, 1H), 6.67-6.95 (m, 2H), 6.20-6.45 (m, 1H), 4.60 (s, 2H).
To a stirred solution of 3-{3-[4-(aminomethyl)phenyl]-5-phenylimidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 4-1) (615 mg, 1.57 mmol, 1 equiv) and 4-bromo-2-fluoropyridine (250 mg, 1.42 mmol, 0.9 equiv) in 1-methyl-2-pyrrolidone (10 mL) was added N,N-diisopropylethylamine (1.1 g, 8.5 mmol, 5.4 equiv). The resulting mixture was stirred overnight at 140° C. The reaction was quenched by the addition of water (10 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (3×10 mL). The combined organic layers were washed with brine (3×10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-30% gradient of ethyl acetate in petroleum ether to afford 3-[3-(4-{[(4-bromopyridin-2-yl)amino]methyl}phenyl)-5-phenylimidazo[4,5-b]pyridin-2-yl]pyridin-2-amine (Intermediate 81-1) (600 mg, 70%) as a brown oil. MS (ESI) calculated for C29H22BrN7: 547.11 m/z, found 548.10 [M+H]+.
To a stirred solution of 4-bromo-2-(1,3-dioxolan-2-yl)phenol (Intermediate 3-2) (5.00 g, 20.4 mmol, 1 equiv) and tert-butyldimethylsilyl chloride (3.5 g, 23 mmol, 1.1 equiv) in dichloromethane (50 mL) was added imidazole (2.00 g, 29.4 mmol). The resulting mixture was stirred overnight at room temperature. The reaction was quenched by the addition of water (50 mL). The resulting mixture was extracted with ethyl acetate (3×50 mL). The combined organic layers were washed with brine (3×50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-5% gradient of ethyl acetate in petroleum ether to afford 4-bromo-2-(1,3-dioxolan-2-yl)phenoxy(tert-butyl)dimethylsilane (5 g, 68%) as yellow oil. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 7.20-7.40 (m, 2H), 6.40-6.80 (m, 1H), 5.60-5.80 (m, 1H), 3.60-3.80 (m, 2H), 3.80-4.00 (m, 2H), 0.70-0.85 (m, 9H).
To a stirred solution of 4-bromo-2-(1,3-dioxolan-2-yl)phenoxy(tert-butyl)dimethylsilane (1.00 g, 2.78 mmol) and bis(pinacolato)diboron (0.85 g, 3.3 mmol) in 1,4-dioxane (4 mL) were added [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (100 mg, 0.137 mmol) and potassium acetate (600 mg, 6.11 mmol) under nitrogen atmosphere. The resulting mixture was stirred overnight at 90° C. The reaction was quenched by the addition of water (10 mL) at room temperature. The mixture was extracted with ethyl acetate (3×10 mL). The combined organic layers were washed with brine (3×10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-40% gradient of ethyl acetate in petroleum ether to afford tert-butyl(2-(1,3-dioxolan-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)dimethylsilane (Intermediate 81-2) as yellow solid. MS (ESI) calculated for C21H35BO5Si: 406.23 m/z, found 407.25 [M+H]+.
Example 82 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 74-1 in place of the amine starting material and Intermediate 69-1 in place of Intermediate 1-4. MS (ESI) calculated for C29H24N6O3: 504.19 m/z, found 505.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.21 (s, 1H), 8.40-8.50 (m, 1H), 8.21-8.42 (m, 1H), 8.00-8.22 (m, 1H), 7.70-7.85 (m, 1H), 7.55-7.70 (m, 1H), 7.38-7.55 (m, 3H), 7.21-7.35 (m, 2H), 6.91-7.12 (m, 2H), 6.70-6.85 (m, 1H), 4.20-4.40 (s, 2H), 1.30-1.55 (m, 2H), 1.02-1.11 (m, 2H).
Example 83 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 74-1 in place of the amine starting material and Intermediate 3-4 in place of Intermediate 1-4. MS (ESI) calculated for C26H20N6O3: 464.16 m/z, found 465.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.28 (s, 1H), 8.30-8.31 (m, 2H), 8.12-8.29 (m, 1H), 7.97-7.98 (m, 2H), 7.35-7.45 (m, 5H), 6.85-7.39 (m, 2H), 6.41-6.44 (m, 1H), 4.39-4.53 (m, 2H).
Example 84 was prepared in a manner analogous to Example 12 using Intermediate 74-1 in place of Intermediate 12-1, 4-formyl-3-hydroxybenzoic acid in place of 3-fluoro-5-formyl-4-hydroxybenzoic acid and PyBOP in place of HATU. MS (ESI) calculated for C26H20N6O3: 464.16 m/z, found 465.10 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.30-10.32 (m, 1H), 8.19-8.31 (m, 2H), 7.99-8.00 (m, 1H), 7.70-7.79 (m, 1H), 7.37-7.56 (m, 7H), 7.37-7.40 (m, 1H), 6.39-6.43 (m, 1H), 4.41-4.54 (m, 2H).
Example 85 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 85-1 in place of Intermediate 1-4. MS (ESI) calculated for C32H23FN6O3: 558.18 m/z, found 559.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.29 (s, 1H), 8.26-8.29 (m, 1H), 7.98-8.04 (m, 4H), 7.38-7.57 (m, 8H), 7.23-7.27 (m, 1H), 7.12-7.16 (m, 1H), 6.44-6.48 (m, 1H), 4.60-4.61 (m, 2H).
Intermediate 85-1 was prepared in a manner analogous to Intermediate 3-4 using Intermediate 21-1 in place of Intermediate 3-3. MS (ESI) calculated for C18H17FO6: 348.10 m/z, found 347.05 [M−H]−.
Example 86 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 86-2 in place of Intermediate 1-4, Intermediate 86-3 in place of the amine starting material and T3P in place of PyBOP. MS (ESI) calculated for C27H22N6O3: 478.18 m/z, found 479.10 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.04 (s, 1H), 8.32-8.34 (m, 1H), 8.18-8.22 (m, 1H), 7.94-7.96 (m, 1H), 7.62-7.64 (m, 1H), 7.51-7.52 (m, 1H), 7.32-7.43 (m, 3H), 7.12-7.19 (m, 2H), 7.03-7.07 (m, 2H), 6.31-6.35 (m, 1H), 2.99-3.01 (m, 2H), 2.75-2.78 (m, 2H).
To a solution of 3-[3-(4-bromophenyl)imidazo[4,5-b]pyridin-2-yl]pyridin-2-amine (Intermediate 86-1) (360 mg, 0.983 mmol, 1 equiv) in N,N-dimethylformamide (5 mL) was added benzyl acrylate (866 mg, 4.92 mmol, 5 equiv), palladium(II) acetate (22 mg, 0.098 mmol, 0.1 equiv), triphenylphosphine (52 mg, 0.20 mmol, 0.20 equiv) and potassium carbonate (271 mg, 1.97 mmol, 2 equiv). The resulting mixture was stirred at 110° C. under nitrogen atmosphere overnight. The mixture was cooled to room temperature, quenched with water (20 mL), and extracted with ethyl acetate (3×20 mL). The organic layers were combined, dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by reverse-phase flash column chromatography on C18 silica gel using a 20-95% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) with a 10 min hold at 70% acetonitrile to afford benzyl (2E)-3-{4-[2-(2-aminopyridin-3-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}prop-2-enoate (250 mg, 56%) as a yellow solid. MS (ESI) calculated for C27H21N5O2: 447.17 m/z, found 448.10 [M+H]+.
To a mixture of benzyl (2E)-3-{4-[2-(2-aminopyridin-3-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}prop-2-enoate (250 mg, 0.559 mmol, 1 equiv) in ethyl acetate (20 mL) was added 10% palladium on carbon (100 mg, 0.15 equiv) and the mixture was stirred under hydrogen atmosphere for 1 h at room temperature. The resulting mixture was filtered and concentrated in vacuo. The residue obtained was purified by silica gel column chromatography using a 0-30% gradient of ethyl acetate in petroleum ether to afford 3-{4-[2-(2-aminopyridin-3-yl)imidazo[4,5-b]pyridin-3-yl]phenyl}propanoic acid (Intermediate 86-2) (130 mg, 62%) as a yellow solid. MS (ESI) calculated for C20H17N5O2: 359.14 m/z, found 360.10 [M+H]+.
Intermediate 86-1 was prepared in a manner analogous to Intermediate 11-1 (using Steps 1 and 2 only) using 2-chloro-3-nitropyridine in place of 2-chloro-3-nitro-6-phenylpyridine and 4-bromoaniline in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C17H12BrN5: 365.03 m/z, found 366.00 [M+H]+.
To a solution of 2-{4-bromo-2-[(4-methoxyphenyl)methoxy]phenyl}-1,3-dioxolane (Intermediate 1-3) (2.00 g, 5.48 mmol, 1 equiv) in 1,4-dioxane (20 mL) were added O-benzyl carbamate (0.99 g, 6.6 mmol, 1.2 equiv), XantPhos (0.63 g, 1.1 mmol, 0.2 equiv) and tris(dibenzylideneacetone)dipalladium(0) (0.50 g, 0.55 mmol, 0.1 equiv). The resulting mixture was stirred under nitrogen atmosphere at 90° C. for 3 h. The resulting mixture was cooled to room temperature and quenched with water (100 mL). The mixture was extracted with ethyl acetate (3×100 mL). The organic layers were combined, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue obtained was purified by silica gel column chromatography using a 0-20% gradient of ethyl acetate in petroleum ether to afford benzyl N-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]carbamate (1.4 g, 59%) as a yellow solid. MS (ESI) calculated for C25H25NO6: 435.17 m/z, found 435.15 [M+H]+.
To a solution of benzyl N-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]carbamate (1.00 g, 2.30 mmol, 1 equiv) in 10 mL of methanol was added palladium on carbon (10%, 110 mg, 0.05 equiv) and the resulting mixture was stirred at room temperature for 1 h under hydrogen atmosphere. The mixture was then filtered and concentrated in vacuo to afford 4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]aniline (Intermediate 86-3) (500 mg, 72%) as a yellow solid. MS (ESI) calculated for C17H19NO4: 301.13 m/z, found 302.15 [M+H]+.
Example 87 was prepared in a manner analogous to Example 81 using Intermediate 87-1 in place of Intermediate 81-2. MS (ESI) calculated for C36H27N7O2: 589.22 m/z, found 590.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.31 (s, 1H), 8.20-8.30 (m, 1H), 8.05-8.15 (m, 1H), 7.90-8.05 (m, 4H), 7.70-7.80 (m, 1H), 7.50-7.60 (m, 2H), 7.30-7.50 (m, 5H), 7.10-7.30 (m, 3H), 6.75-6.95 (m, 2H), 6.30-6.40 (m, 1H), 4.66 (s, 2H).
Intermediate 87-1 was prepared in a manner analogous to Intermediate 81-2 using Intermediate 1-2 in place of Intermediate 3-2. MS (ESI) calculated for C21H35BO5Si: 406.23 m/z, found 407.23 [M+H]+.
Example 88 was prepared in a manner analogous to Example 19 using Intermediate 67-1 in place of Intermediate 19-2, Intermediate 88-1 in place of Intermediate 19-3 and dichloromethane/2,2,2-trifluoroacetic acid (15:1) in place of 2,2,2-trifluoroacetic aacid/methanesulfonic acid. MS (ESI) calculated for C32H26FN7O2: 559.61 m/z, found 560.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.20 (s, 1H), 9.20-9.22 (m, 1H), 8.58-8.59 (m, 1H), 8.40-8.41 (m, 1H), 8.32-8.38 (m, 1H), 8.09-8.13 (m, 1H), 7.68-8.07 (m, 1H), 7.50-7.66 (m, 4H), 7.48-7.50 (m, 2H), 7.27-7.29 (m, 1H), 6.92-6.95 (m, 1H), 6.42-6.45 (m, 1H), 4.30-4.31 (m, 2H), 3.22-3.26 (m, 2H), 3.04-3.17 (m, 2H). 19F-NMR (400 MHz, DMSO-d6) δ (ppm): −140.30.
Intermediate 88-1 was prepared in a manner analogous to Intermediate 16-4 using Intermediate 21-1 in place of Intermediate 16-3. MS (ESI) calculated for C19H19FO5: 346.12 m/z, found 347.15 [M+H]+.
Example 89 was prepared in a manner analogous to Example 19 using Intermediate 74-1 in place of Intermediate 19-2, Intermediate 89-2 in place of Intermediate 19-3, and dichloromethane/2,2,2-trifluoroacetic acid (15:1) in place of 2,2,2-trifluoroacetic acid/methanesulfonic acid. MS (ESI) calculated for C29H26FN7O2: 523.21 m/z, found 524.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 9.85 (s, 1H), 8.31-8.32 (m, 1H), 8.18-8.20 (m, 1H), 7.96-7.97 (m, 1H), 7.52-7.54 (m, 1H), 7.33-7.44 (m, 6H), 7.16-7.18 (m, 1H), 6.33-6.36 (m, 1H), 3.77 (s, 2H), 2.72-2.87 (m, 4H), 2.10 (s, 3H). 19F NMR (376 MHz, DMSO-d6) (ppm): −127.32.
To a solution of N-[3-bromo-6-(1,3-dioxolan-2-yl)-2-fluorophenyl]acetamide (Intermediate 89-1) (1.2 g, 3.9 mmol, 1 equiv) in N,N-dimethylformamide (10 mL) were added tributyl(prop-2-en-1-yl)stannane (2.61 g, 7.89 mmol, 2 equiv) and bis(triphenylphosphine)palladium(II) dichloride (0.28 g, 0.40 mmol, 0.1 equiv) and the resulting mixture was stirred for 1 h at 80° C. The resulting mixture was cooled to room temperature and purified by reverse-phase flash column chromatography on C18 silica gel using a 5-65% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) to afford N-[6-(1,3-dioxolan-2-yl)-2-fluoro-3-(prop-2-en-1-yl)phenyl]acetamide (800 mg, 76%) a yellow oil. MS (ESI) calculated for C14H16FNO3: 265.11 m/z, found 266.15 [M+H]+.
To a solution of N-[6-(1,3-dioxolan-2-yl)-2-fluoro-3-(prop-2-en-1-yl)phenyl]acetamide (800 mg, 3.02 mmol, 1 equiv) in tetrahydrofuran (10 mL) and water (10 mL) were added osmium tetroxide (767 mg, 3.02 mmol, 1 equiv) and sodium periodate (1.29 g, 6.03 mmol, 2 equiv) and the resulting mixture was stirred at room temperature for 1 h. The resulting mixture was extracted with ethyl acetate (10 mL×3). The combined organic phases were washed with brine (30 mL×2), dried with sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography to afford N-[6-(1,3-dioxolan-2-yl)-2-fluoro-3-(2-oxoethyl)phenyl]acetamide (Intermediate 89-2) (480 mg, 60%) as a yellow oil. MS (ESI) calculated for C13H14FNO4: 267.09 m/z, found 268.15 [M+H]+.
To a cooled (0° C.) solution of 2-amino-4-bromo-3-fluorobenzoic acid (10.0 g, 42.7 mmol, 1 equiv) in tetrahydrofuran (500 mL) was added lithium aluminum hydride (3.24 g, 85.5 mmol, 2 equiv) and the resulting mixture was stirred at room temperature for 1 h. The reaction was quenched by sequential addition of water (86 mL), sodium hydroxide (2 M, 86 mL) and water (86 mL). The resulting mixture was filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography to afford (2-amino-4-bromo-3-fluorophenyl)methanol (5 g, 53%) as a yellow oil. MS (ESI) calculated for C7H5BrFNO: 218.97 m/z, found 217.85 [M−H]−.
To a solution of (2-amino-4-bromo-3-fluorophenyl)methanol (5.00 g, 22.7 mmol, 1 equiv) in 1,2-dichloroethane (300 mL) was added manganese (IV) oxide (29.63 g, 340.8 mmol, 15 equiv) and the resulting mixture was stirred for 3 h at 50° C. The resulting mixture was filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography to afford 2-amino-4-bromo-3-fluorobenzaldehyde (3.2 g, 65%) as a yellow oil. MS (ESI) calculated for C7H5BrFNO: 216.95 m/z, found 215.90 [M−H]−.
To a cooled (0° C.) solution of 2-amino-4-bromo-3-fluorobenzaldehyde (3.00 g, 13.8 mmol, 1 equiv) in dichloromethane (300 mL) were added pyridine (5.44 g, 68.8 mmol, 5 equiv) and acetyl chloride (5.40 g, 68.8 mmol, 5 equiv) and the resulting mixture was stirred at room temperature for 1 h. The reaction was diluted with water (100 mL) and extracted with ethyl acetate (3×200 mL). The combined organic layers were washed with water (3×100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase flash column chromatography on C18 silica gel using a 5-70% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) to afford N-(3-bromo-2-fluoro-6-formylphenyl)acetamide (2.3 g, 60%) as a yellow oil. MS (ESI) calculated for C9H7BrFNO2: 258.96 m/z, found 257.90 [M−H]−.
To a solution of N-(3-bromo-2-fluoro-6-formylphenyl)acetamide (2.3 g, 8.8 mmol, 1 equiv), in toluene (30 mL) were added p-toluenesulfonic acid (0.15 g, 0.88 mmol, 0.1 equiv), triethyl orthoformate (6.55 g, 44.2 mmol, 5 equiv) and ethylene glycol (1.65 g, 26.5 mmol, 3 equiv) and the resulting mixture was stirred for 18 h at 90° C. The resulting mixture was cooled to room temperature, concentrated in vacuo, and purified by silica gel column chromatography to afford N-[3-bromo-6-(1,3-dioxolan-2-yl)-2-fluorophenyl]acetamide (Intermediate 89-1) (1.46 g, 54%) as a white solid. MS (ESI) calculated for C11H11BrFNO3: 302.99 m/z, found 303.95 [M+H]+.
Example 90 was prepared in a manner analogous to Example 19 using Intermediate 4-1 in place of Intermediate 19-2, Intermediate 89-2 in place of Intermediate 19-3, and dichloromethane/2,2,2-trifluoroacetic acid (6:1) in place of 2,2,2-trifluoroacetic acid/methanesulfonic acid. MS (ESI) calculated for C35H30FN7O2: 599.24 m/z, found 600.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 9.90 (s, 1H), 8.26-8.32 (m, 1H), 7.97-8.06 (m, 4H), 7.55-7.68 (m, 5H), 7.36-7.50 (m, 4H), 7.20-7.26 (m, 1H), 6.40 (dd, J=7.6, 4.9 Hz, 1H), 4.26 (s, 2H), 3.18-3.25 (m, 2H), 3.04-3.12 (m, 2H), 2.14 (s, 3H). 19F NMR (376 MHz, DMSO-d6) (ppm): −126.27.
Example 91 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 74-1 in place of the amine starting material and Intermediate 91-1 in place of Intermediate 1-4. MS (ESI) calculated for C29H25N7O2: 519.20 m/z, found 520.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.73 (s, 1H), 9.93 (s, 1H), 8.77-8.76 (m, 1H), 8.31-8.32 (m, 1H), 8.19-8.21 (m, 1H), 8.10-8.14 (m, 1H), 7.99-8.00 (m, 1H), 7.78-7.80 (m, 1H), 7.37-7.40 (m, 5H), 7.20-7.25 (m, 2H), 7.02-7.21 (m, 2H), 6.40-6.43 (m, 1H), 4.36-4.38 (m, 2H), 3.56-3.61 (m, 2H), 2.07-2.14 (m, 3H).
To a solution of 2-amino-4-bromobenzaldehyde (4.95 g, 24.9 mmol, 1 equiv) in toluene (100 mL) was added p-toluenesulfonic acid (0.43 g, 2.5 mmol, 0.1 equiv), ethylene glycol (7.72 g, 124 mmol, 5 equiv) and triethyl orthoformate (11.06 g, 74.62 mmol, 3 equiv). The obtained solution was stirred at room temperature for 10 min then for 18 h at 90° C. The resulting mixture was cooled to 0° C. and quenched by the addition of saturated aqueous ammonium chloride (80 mL). The mixture was extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with water (100 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-7% gradient of ethyl acetate in petroleum ether to provide 5-bromo-2-(1,3-dioxolan-2-yl)aniline (3 g, 49%) as a light-yellow oil. MS (ESI) calculated for C9H10BrNO2: 242.99 m/z, found 244.10 [M+H]+.
A solution of 5-bromo-2-(1,3-dioxolan-2-yl)aniline (3.00 g, 12.3 mmol, 1 equiv) in pyridine was sparged with argon for 15 min under dry conditions. Acetyl chloride (3.86 g, 49.2 mmol, 4 equiv) was added and the resulting mixture stirred for 2 h at room temperature. The residue was purified by silica gel column chromatography to afford N-(5-bromo-2-(1,3-dioxolan-2-yl)phenyl)acetamide (2 g, 57%) as a yellow oil. MS (ESI) calculated for C11H12BrNO3: 285.00 m/z, found 286.05, 288.05 [M+H, M+H+2]+.
To a solution of N-[5-bromo-2-(1,3-dioxolan-2-yl)phenyl]acetamide (2.0 g, 7.0 mmol, 1 equiv) and (2-ethoxy-2-oxoethyl)zinc(II) bromide (20 mL, 0.5 M in tetrahydrofuran, 1.5 equiv) in tetrahydrofuran (5 mL) were added [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.50 g, 0.70 mmol, 0.1 equiv) and XPhos (0.65 g, 1.4 mmol, 0.2 equiv). After stirring overnight at 100° C. under a nitrogen atmosphere, the mixture was cooled to 0° C. and quenched with water (10 mL). The resulting mixture was extracted with ethyl acetate (20 mL×3). The combined organic layers were washed with water (10 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-60% gradient of ethyl acetate in petroleum ether to provide ethyl 2-(3-acetamido-4-(1,3-dioxolan-2-yl)phenyl)acetate (1.0 g, 49%) as a light-yellow solid. MS (ESI) calculated for C15H19NO5: 293.13 m/z, found 294.10 [M+H]+.
To a solution of ethyl 2-(3-acetamido-4-(1,3-dioxolan-2-yl)phenyl)acetate (1.0 g, 3.4 mmol, 1 equiv) in tetrahydrofuran (10 mL) and water (10 mL) was added lithium hydroxide (5.0 mL, 2 M in water, 3 equiv). The resulting mixture was stirred at room temperature for 1 h then concentrated in vacuo and purified by reverse-phase flash column chromatography on C18 silica gel using a 20-95% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) with a 10 min hold at 70% acetonitrile to provide 2-(3-acetamido-4-(1,3-dioxolan-2-yl)phenyl)acetic acid (Intermediate 91-1) (600 mg, 66%). MS (ESI) calculated for C13H15NO5: 265.10 m/z, found 266.10 [M+H]+.
Example 92 was prepared in a manner analogous to Example 19 using Intermediate 92-2 in place of Intermediate 19-2, Intermediate 92-3 in place of Intermediate 19-3 and dichloromethane/2,2,2-trifluoroacetic acid (5:1) in place of 2,2,2-trifluoroacetic acid/methanesulfonic acid. MS (ESI) calculated for C34H30N6O2: 554.24 m/z, found 555.30 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.22 (s, 1H), 8.23-8.29 (m, 1H), 7.95-8.02 (m, 5H), 7.56-7.57 (m, 2H), 7.64-7.66 (m, 1H), 7.33-7.48 (m, 8H), 7.09-7.12 (m, 1H), 6.21-6.25 (m, 1H), 4.04-4.06 (m, 1H), 2.71-2.74 (m, 3H), 2.63 (s, 1H), 1.41-1.49 (m, 3H).
Intermediate 92-2 was prepared in a manner analogous to Intermediate 11-1 using Intermediate 92-1 in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C25H22N6: 406.19 m/z, found 407.10 [M+H]+.
To a solution of (S)-1-(4-nitrophenyl)ethan-1-amine (5.00 g, 30.1 mmol, 1 equiv) in dichloromethane (100 mL) were added di-tert-butyl dicarbonate (13.13 g, 60.18 mmol, 2 equiv) and triethylamine (9.13 g, 90.3 mmol, 3 equiv). The resulting mixture was stirred at room temperature overnight. The reaction was quenched with water (100 mL). The resulting mixture was extracted with dichloromethane (3×100 mL). The organic layers were combined, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue obtained was purified by silica gel column chromatography using a 0-20% gradient of ethyl acetate in petroleum ether to afford tert-butyl (S)-(1-(4-nitrophenyl)ethyl)carbamate (6 g, 75%) as a white solid. MS (ESI) calculated for C13H18N2O4: 266.13 m/z, found 267.15 [M+H]+.
To a solution of tert-butyl N-[(1S)-1-(4-nitrophenyl)ethyl]carbamate (1.00 g, 3.756 mmol, 1 equiv) in methanol (10 mL) and water (10 mL) were added iron (1.68 g, 30.0 mmol, 8 equiv) and ammonium chloride (1.00 g, 18.8 mmol, 5 equiv). The resulting mixture was stirred at 70° C. for 1 h. The resulting mixture was cooled to room temperature and quenched with water (50 mL). The mixture was extracted with ethyl acetate (3×50 mL). The organic layers were combined, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue obtained was purified by silica gel column chromatography using a 0-10% gradient of ethyl acetate in petroleum ether to afford tert-butyl N-[(1S)-1-(4-aminophenyl)ethyl]carbamate (Intermediate 92-1) (850 mg, 96%) as a yellow solid. MS (ESI) calculated for C13H20N2O2: 236.15 m/z, found 237.25 [M+H]+.
To a solution of 2-(5-bromo-2-((4-methoxybenzyl)oxy)phenyl)-1,3-dioxolane (Intermediate 3-3) (3 g, 8.2 mmol, 1 equiv) and tributyl(prop-2-en-1-yl)stannane (5.44 g, 16.4 mmol, 2 equiv) in N,N-dimethylformamide (24 mL) was added bis(triphenylphosphine)palladium(II) dichloride (0.58 g, 0.82 mmol, 0.1 equiv). After stirring for 1 h at 80° C. under a nitrogen atmosphere, the mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography on C18 silica gel using a 10 to 40% gradient of acetonitrile to provide 2-(5-allyl-2-((4-methoxybenzyl)oxy)phenyl)-1,3-dioxolane (2.4 g, 90%) as colorless oil. MS (ESI) calculated for C20H22O4: 326.15 m/z, found 327.20 [M+H]+.
To a solution of 2-(5-allyl-2-((4-methoxybenzyl)oxy)phenyl)-1,3-dioxolane (0.5 g, 1.53 mmol, 1 equiv) in acetonitrile (3 mL) and water (3 mL) were added osmium tetroxide (1.17 g, 4.60 mmol, 3 equiv) and sodium periodate (0.98 g, 4.60 mmol, 3 equiv). After stirring for 30 min at room temperature, the reaction was quenched by the addition of water. The resulting mixture was extracted with ethyl acetate (20 mL×3). The combined organic layers were washed with water (20 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography using a gradient of 0-25% ethyl acetate in petroleum ether to provide 2-(3-(1,3-dioxolan-2-yl)-4-((4-methoxybenzyl)oxy)phenyl) acetaldehyde (Intermediate 92-3) (0.15 g, 30%) as a light-yellow oil. MS (ESI) calculated for C19H20O5: 328.13 m/z, found 329.15 [M+H]+.
Example 93 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 74-1 in place of the amine starting material and Intermediate 21-2 in place of Intermediate 1-4. MS (ESI) calculated for C27H21FN6O3: 496.17 m/z, found 497.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.15 (s, 1H), 8.30-8.32 (m, 1H), 8.21-8.29 (m, 1H), 7.95-7.97 (m, 1H), 7.39-7.43 (m, 6H), 7.22-7.24 (m, 1H), 6.86-6.90 (m, 1H), 6.40-6.44 (m, 1H), 4.35-4.36 (m, 2H), 3.63-3.72 (m, 2H). 19F-NMR (400 MHz, DMSO-d6) δ (ppm): −139.35.
Example 94 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 21-2 in place of Intermediate 1-4. MS (ESI) calculated for C33H25FN6O3: 572.20 m/z, found 573.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.15 (s, 1H), 8.31-8.33 (m, 1H), 7.99-8.04 (m, 4H), 7.79-7.99 (m, 1H), 7.39-7.77 (m, 8H), 6.82-6.95 (m, 2H), 4.20-4.60 (m, 2H), 3.62-3.74 (m, 2H). 19F-NMR (400 MHz, DMSO-d6) δ (ppm): −139.30.
Example 95 was prepared in a manner analogous to Example 19 using Intermediate 74-1 in place of Intermediate 19-2, Intermediate 88-1 in place of Intermediate 19-3 and dichloromethane/2,2,2-trifluoroacetic acid (15:1) in place of 2,2,2-trifluoroacetic acid/methanesulfonic acid. MS (ESI) calculated for C27H23FN6O2: 482.19 m/z, found 483.20 [M+H]+. 1H-NMR (300 MHz, DMSO-d6) δ (ppm): 10.12-10.19 (m, 1H), 8.30-8.34 (m, 1H), 8.20-8.22 (m, 2H), 7.96-8.00 (m, 1H), 7.52-7.55 (m, 2H), 7.40-7.43 (m, 5H), 7.18-7.23 (m, 1H), 6.83-6.87 (m, 1H), 6.36-6.41 (m, 1H), 4.00-4.01 (m, 2H), 2.84-2.93 (m, 4H).
Example 96 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 74-1 in place of the amine starting material. MS (ESI) calculated for C27H22N6O3: 478.18 m/z, found 479.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.19 (s, 1H), 8.33-8.35 (m, 1H), 8.14-8.23 (m, 1H), 8.00-8.02 (m, 1H), 7.60-7.62 (m, 1H), 7.33-7.43 (m, 6H), 6.89-6.96 (m, 2H), 6.46-6.51 (m, 1H), 4.30-4.36 (m, 2H), 3.46-3.54 (m, 2H).
Example 97 was prepared in a manner analogous to Example 19 using Intermediate 4-1 in place of Intermediate 19-2, Intermediate 97-2 in place of Intermediate 19-3 and dichloromethane/2,2,2-trifluoroacetic acid (10:1) in place of 2,2,2-trifluoroacetic acid/methanesulfonic acid. MS (ESI) calculated for C34H30N6O2: 554.24 m/z, found 555.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.25 (s, 1H), 8.24-8.26 (m, 1H), 7.99-8.01 (m, 4H), 7.97-7.98 (m, 1H), 7.50-7.61 (m, 7H), 7.19-7.36 (m, 1H), 7.08-7.16 (m, 1H), 7.69-7.08 (m, 1H), 6.33-6.36 (m, 1H), 3.86-3.87 (m, 3H), 3.54-3.57 (m, 2H), 2.76-2.85 (m, 4H).
Intermediate 97-2 was prepared in a manner analogous to Intermediate 16-4 using Intermediate 97-1 in place of Intermediate 16-3. MS (ESI) calculated for C12H14O4: 222.09 m/z, found 223.15 [M+H]+.
Intermediate 97-1 was prepared in a manner analogous to Intermediate 1-2 using 4-bromo-2-methoxybenzaldehyde in place of 4-bromo-2-hydroxybenzaldehyde. MS (ESI) calculated for C10H11BrO3: 257.99 m/z, found 259.05 [M+H]+.
Example 98 was prepared in a manner analogous to Example 19 using Intermediate 74-1 in place of Intermediate 19-2, Intermediate 98-3 in place of Intermediate 19-3 and dichloromethane/2,2,2-trifluoroacetic acid (10:1) in place of 2,2,2-trifluoroacetic acid/methanesulfonic acid. MS (ESI) calculated for C29H27N7O2: 505.22 m/z, found 506.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 9.94 (s, 1H), 8.32-8.40 (m, 1H), 8.19-8.28 (m, 1H), 8.08-8.15 (m, 1H), 7.96-8.04 (m, 1H), 7.81-7.89 (m, 1H), 7.59-7.68 (m, 2H), 7.49-7.57 (m, 2H), 7.39-7.48 (m, 2H), 7.18-7.27 (m, 1H), 6.47-6.59 (m, 1H), 4.27 (s, 2H), 3.19-3.27 (m, 2H), 3.00-3.08 (m, 2H), 2.15 (s, 3H).
Intermediate 98-3 was prepared in a manner analogous to Intermediate 16-4 using Intermediate 98-2 in place of Intermediate 16-3. MS (ESI) calculated for C13H15NO4: 249.10 m/z, found 250.10 [M+H]+.
Step 1: Synthesis of N-[5-bromo-2-(1,3-dioxolan-2-yl)phenyl]acetamide (Intermediate 98-1)
To a solution of 5-bromo-2-(1,3-dioxolan-2-yl)aniline (Intermediate 98-1) (9.00 g, 36.9 mmol, 1 equiv) in pyridine (180 mL) was added acetyl chloride (11.58 g, 147.5 mmol, 4 equiv) and the resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated in vacuo and the residue was purified by silica gel column chromatography to afford N-[5-bromo-2-(1,3-dioxolan-2-yl)phenyl]acetamide (Intermediate 98-2) (2 g, 19%) as a yellow oil. MS (ESI) calculated for C11H12BrNO3: 285.00 m/z, found 286.05, 288.05 [M+H, M+H+2]+.
To a solution of 4-bromo-2-nitrobenzaldehyde (5.00 g, 21.7 mmol, 1 equiv) in toluene (50 mL) were added ethylene glycol (6.75 g, 109 mmol, 5 equiv), triethyl orthoformate (9.66 g, 65.2 mmol, 3 equiv) and p-toluenesulfonic acid (0.37 g, 2.2 mmol, 0.1 equiv). The resulting mixture was stirred at 90° C. for 18 h under nitrogen atmosphere. The reaction was cooled to room temperature and quenched with water (30 mL), which was added dropwise over 10 min. The resulting mixture was extracted with dichloromethane (30 mL×2), and the combined extracts were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography to afford 2-(4-bromo-2-nitrophenyl)-1,3-dioxolane (1.89 g, 32%) as a yellow oil. MS (ESI) calculated for C9HsBrNO4: 272.96 m/z, found 273.90 [M+H]+.
To a solution of 2-(4-bromo-2-nitrophenyl)-1,3-dioxolane (650 mg, 2.37 mmol, 1 equiv) in methanol (10 mL) were added iron (1.32 g, 23.7 mmol, 10 equiv) and ammonium chloride (634 mg, 11.9 mmol, 5 equiv). The resulting mixture was stirred at 60° C. for 3 h. The mixture was then cooled to room temperature and filtered, rinsing with ethyl acetate. The filtrate was concentrated, and the residue was taken up into water (30 mL). The resulting mixture was extracted with ethyl acetate (3×30 mL). The organic layers were combined, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue obtained was purified by silica gel chromatography using a 0-30% gradient of ethyl acetate in petroleum ether to afford 5-bromo-2-(1,3-dioxolan-2-yl)aniline (Intermediate 98-1) (300 mg, 52%) as a yellow solid. MS (ESI) calculated for C9H10BrNO2: 242.99 m/z, found 243.98 [M+H]+.
Example 99 was prepared in a manner analogous to Example 19 using Intermediate 99-1 in place of Intermediate 19-2, Intermediate 16-4 in place of Intermediate 19-3 and tetrahydrofuran/triethylamine trihydrofluoride (10:1) in place of 2,2,2-trifluoroacetic acid/methanesulfonic acid. MS (ESI) calculated for C27H23ClN6O2: 498.16 m/z, found 499.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.19 (s, 1H), 8.32-8.35 (m, 1H), 8.04-8.05 (m, 1H), 7.52-7.69 (m, 8H), 6.90-6.92 (m, 2H), 6.67-6.68 (m, 1H), 4.28-4.29 (m, 2H), 3.25-3.27 (m, 2H), 2.97-2.98 (m, 2H).
Intermediate 99-1 was prepared in a manner analogous to Intermediate 24-2 using Intermediate 1-1 in place of Intermediate 24-1. MS (ESI) calculated for C18H15ClN6: 350.10 m/z, found 351.05 [M+H]+.
Example 100 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 100-1 in place of the amine starting material and Intermediate 100-2 in place of Intermediate 1-4. MS (ESI) calculated for C34H28N6O3: 568.22 m/z, found 569.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.16 (s, 1H), 8.27-8.29 (m, 1H), 8.01-8.03 (m, 3H), 7.91-7.98 (m, 1H), 7.42-7.58 (m, 8H), 7.10-7.13 (m, 1H), 6.71-6.77 (m, 2H), 6.10 (s, 1H), 3.20-3.26 (m, 3H), 2.75-2.89 (m, 2H), 2.36-2.52 (m, 2H).
Intermediate 100-1 was prepared in a manner analogous to Intermediate 11-1 using tert-butyl N-(4-aminophenyl)-N-methylcarbamate in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C24H20N6: 392.17 m/z, found 393.25 [M+H]+.
Intermediate 100-2 was prepared in a manner analogous to Intermediate 86-2 using Intermediate 1-3 in place of Intermediate 86-1. MS (ESI) calculated for C20H22O6: 358.14 m/z, found 359.10 [M+H]+.
Example 101 was prepared in a manner analogous to Example 19 using Intermediate 101-1 in place of Intermediate 19-2, Intermediate 16-4 in place of Intermediate 19-3 and tetrahydrofuran/triethylamine trihydrofluoride (10:1) in place of 2,2,2-trifluoroacetic acid/methanesulfonic acid. MS (ESI) calculated for C34H31N7O4S: 633.22 m/z, found 634.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.17 (s, 1H), 8.27 (s, 1H), 7.89-7.98 (m, 3H), 7.53-7.72 (m, 5H), 7.41 (s, 2H), 7.23 (s, 2H), 6.85-6.88 (m, 2H), 6.39 (s, 1H), 4.07-4.31 (m, 2H), 3.22-3.25 (s, 3H), 2.78-3.00 (m, 4H).
Intermediate 101-1 was prepared in a manner analogous to Intermediate 12-1 using 3-methanesulfonamidophenylboronic acid in place of the boronic acid. MS (ESI) calculated for C25H23N7O2S: 485.16 m/z, found 486.15 [M+H]+.
Example 102 was prepared in a manner analogous to Example 19 using Intermediate 102-1 in place of Intermediate 19-2 and Intermediate 88-1 in place of Intermediate 19-3. MS (ESI) calculated for C35H30FN7O3: 615.24 m/z, found 616.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.17 (s, 1H), 8.28-8.31 (m, 2H), 8.01-8.22 (m, 1H), 7.94-7.99 (m, 1H), 6.93-7.03 (m, 2H), 7.65-7.71 (m, 6H), 7.59-7.62 (m, 2H), 7.41-7.56 (m, 1H), 6.45-6.46 (m, 1H), 6.41-6.43 (m, 1H), 4.18 (s, 2H), 3.10-3.11 (m, 2H), 2.98 -2.99 (m, 2H), 2.05-2.08 (m, 3H). 19F NMR (400 MHz, DMSO-d6) δ (ppm): −140.71.
Intermediate 102-1 was prepared in a manner analogous to Intermediate 12-1 using 3-acetamidophenylboronic acid in place of the boronic acid. MS (ESI) calculated for C26H23N7O: 449.20 m/z, found 450.20 [M+H]+.
Example 103 was prepared in a manner analogous to Example 16 using Intermediate 103-1 in place of Intermediate 16-2. MS (ESI) calculated for C32H27N7O2: 541.22 m/z, found 542.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.10-10.21 (m, 1H), 8.67-8.74 (m, 1H), 8.43-8.54 (m, 1H), 8.34-8.43 (m, 1H), 8.18-8.24 (m, 1H), 8.00-8.09 (m, 1H), 7.86-7.96 (m, 1H), 7.63-7.75 (m, 5H), 7.54-7.60 (m, 1H), 7.44-7.52 (m, 1H), 6.90-6.99 (m, 2H), 6.66-6.70 (m, 1H), 4.28-4.36 (m, 2H), 3.23-3.33 (m, 2H), 2.98-3.06 (m, 2H).
Step 1: Synthesis of 3-(3-(4-(aminomethyl)phenyl)-5-(pyridin-2-yl)-3H-imidazo[4,5-b]pyridin-2-yl)pyridin-2-amine (Intermediate 103-1)
tert-Butyl (4-(2-(2-aminopyridin-3-yl)-5-(pyridin-2-yl)-3H-imidazo[4,5-b]pyridin-3-yl)benzyl)carbamate (Intermediate 62-1) (150 mg, 0.304 mmol, 1 equiv) was dissolved in dichloromethane (5 mL) and 2,2,2-trifluoroacetic acid (1 mL) was added. The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated in vacuo and the residue was purified by reverse-phase flash column chromatography using a 6-60% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) to afford 3-{3-[4-(aminomethyl)phenyl]-5-(pyridin-2-yl)imidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 103-1) (100 mg, 84%) as a yellow solid. MS (ESI) calculated for C23H19N7: 393.17 m/z, found 394.15 [M+H]+.
To a solution of 4-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl)benzoic acid (Intermediate 29-2) (1.0 g, 2.5 mmol, 1 equiv) in N,N-dimethylformamide (30 mL) were added 2-propynylamine (0.20 g, 3.7 mmol, 1.5 equiv), N,N-diisopropylethylamine (0.95 g, 7.4 mmol, 3.0 equiv), and HATU (1.03 g, 2.70 mmol, 1.1 equiv). The resulting solution was stirred at room temperature for 2 h. The reaction was quenched by the addition of saturated aqueous ammonium chloride (20 mL) at 0° C. The aqueous layer was extracted with ethyl acetate (3×20 mL). The combined organic layers were concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with methanol/dichloromethane (1:20) to afford 4-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl)-N-(prop-2-yn-1-yl)benzamide (500 mg, 45.83%) as a yellow solid. MS (ESI) calculated for C27H20N6O: 444.17 m/z, found 445.20 [M+H]+.
A mixture of 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]-N-(prop-2-yn-1-yl)benzamide (250 mg, 0.562 mmol, 1 equiv), 4-bromo-2-hydroxybenzaldehyde (226 mg, 1.12 mmol, 2.0 equiv), bis(triphenylphosphine)palladium(II) dichloride (39 mg, 0.056 mmol, 0.1 equiv), and copper (I) iodide (11 mg, 0.056 mmol, 0.1 equiv) was dissolved in degassed N,N-diisopropylethylamine (218 mg, 1.69 mmol, 3 equiv) and N,N-dimethylformamide (5 mL) under nitrogen atmosphere. The resulting solution was stirred at 50° C. for 2 h. The reaction was quenched with saturated aqueous ammonium chloride (10 mL). The mixture was extracted with ethyl acetate (3×20 mL) and the combined organic layers were concentrated under reduced pressure. The crude product was purified by Preparative HPLC on a XBridge Shield RP18 OBD Column using a 52-73% gradient of acetonitrile in water (+0.1% formic acid) to afford 4-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl)-N-(3-(4-formyl-3-hydroxyphenyl)prop-2-yn-1-yl)benzamide (Example 104) (45 mg, 14%) as a yellow solid. MS (ESI) calculated for C34H24N6O3: 564.19 m/z, found 565.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 11.32 (br s, 1H), 10.24 (s, 1H), 10.07 (s, 1H), 9.10 (s, 1H), 8.93-8.95 (m, 1H), 8.76-8.78 (m, 1H), 8.36-8.38 (m, 1H), 8.22-8.24 (m, 1H), 8.17-8.19 (m, 2H), 7.92-7.94 (m, 2H), 7.84-7.86 (m, 2H), 7.73-7.75 (m, 1H), 7.55-7.61 (m, 3H), 7.48-7.52 (m, 1H), 7.24-7.28 (m, 1H), 7.00 (s, 1H), 6.92-6.94 (m, 1H), 4.34-4.35 (m, 2H).
Example 105 was prepared in a manner analogous to Example 92 (via Intermediates 92-2 and 92-1) starting from of (R)-1-(4-nitrophenyl)ethan-1-amine in place of (S)-1-(4-nitrophenyl)ethan-1-amine. MS (ESI) calculated for C34H30N6O2: 554.25 m/z, found 555.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.23 (s, 1H), 9.10 (s, 1H), 8.78-8.79 (m, 1H), 8.38-8.42 (m, 1H), 8.20-8.23 (m, 1H), 8.13-8.15 (m, 2H), 7.84-7.87 (m, 2H), 7.68-7.70 (m, 1H), 7.49-7.54 (m, 6H), 7.36-7.39 (m, 1H), 7.27-7.31 (m, 1H), 6.94-6.97 (m, 1H), 4.36-4.41 (m, 1H), 3.06-3.09 (m, 1H), 2.87-2.90 (m, 3H), 1.58-1.60 (m, 3H).
To a solution of 3-(3-(4-bromophenyl)-5-phenyl-3H-imidazo[4,5-b]pyridin-2-yl)pyridin-2-amine (Intermediate 29-1) (0.44 g, 1.0 mmol, 1 equiv) in 1,4-dioxane (10 mL) were added palladium(II) acetate (17 mg, 0.10 mmol, 0.1 equiv), benzyl thiol (541 mg, 5.0 mmol, 5.0 equiv) and XPhos (95 mg, 0.20 mmol, 0.2 equiv). After stirring for 2 h at 80° C. under nitrogen atmosphere, the reaction was cooled to 0° C. and quenched by the addition of water (10 mL). The resulting mixture was extracted with ethyl acetate (20 L×3). The combined organic layers were washed with water (10 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by reverse-phase flash column chromatography on C18 silica gel using a 20-95% gradient of acetonitrile in water (+0.05% ammonium bicarbonate) to afford 3-(3-(4-(benzylthio)phenyl)-5-phenyl-3H-imidazo[4,5-b]pyridin-2-yl)pyridin-2-amine (300 mg, 62%). MS (ESI) calculated for C30H23N5S: 485.17 m/z, found 486.10 [M+H]+.
To a solution of 3-{3-[4-(benzylsulfanyl)phenyl]-5-phenylimidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (200 mg, 0.412 mmol, 1 equiv) in dichloromethane (1.5 mL), acetic acid (0.2 mL) and water (0.1 mL) was added 1,3-dichloro-5,5-dimethylimidazolidine-2,4-dione (243 mg, 1.24 mmol, 3 equiv) and the resulting mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated in vacuo and the residue was purified by silica gel column chromatography using a 0-10% gradient of methanol in dichloromethane to provide 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]benzenesulfonyl chloride (160 mg, 84%) as a yellow solid. MS (ESI) calculated for C23H16ClN5O2S: 461.07 m/z, found 461.95 [M+H]+.
To a solution of 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]benzenesulfonyl chloride (150 mg, 0.325 mmol, 1 equiv) in dichloromethane (3 mL) was added 2-{4-[(tert-butyldimethylsilyl)oxy]-3-(1,3-dioxolan-2-yl)phenyl}ethanamine (Intermediate 106-1) (105 mg, 0.325 mmol, 1 equiv). The resulting mixture was stirred at room temperature for 2 h. After concentration, the residue was purified by reverse-phase flash column chromatography using a 10-95% gradient of acetonitrile in water (+0.05% 2,2,2-trifluoroacetic acid) to afford 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]-N-(2-{3-[(tert-butyldimethylsilyl) oxy]-4-(1,3-dioxolan-2-yl)phenyl}ethyl)benzenesulfonamide (50 mg, 21%) as a yellow solid. MS (ESI) calculated for C40H44N6O5SSi: 748.29 m/z, found 747.10 [M−H]−.
To a solution of 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]-N-(2-{3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)phenyl}ethyl)benzenesulfonamide (50 mg, 0.067 mmol, 1 equiv) in 1,4-dioxane (5 mL) was added triethylamine trihydrofluoride (108 mg, 0.670 mmol, 10 equiv). The resulting mixture was stirred at room temperature for 30 min. The mixture was then concentrated in vacuo and purified by preparative HPLC on a XBridge Prep Phenyl OBD Column using a 28-48% gradient of acetonitrile in water to afford 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]-N-[2-(4-formyl-3-hydroxyphenyl) ethyl]benzenesulfonamide (Example 106) (6.9 mg, 17%) as a light-yellow solid. MS (ESI) calculated for C32H26N6O4S: 590.17 m/z, found 591.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 9.85 (s, 1H), 8.32-8.41 (m, 1H), 8.04-8.15 (m, 6H), 7.81-7.92 (m, 3H), 7.64-7.73 (m, 1H), 7.40-7.60 (m, 5H), 6.78-6.89 (m, 1H), 2.95-3.17 (m, 4H).
A mixture of 5-bromo-2-(1,3-dioxolan-2-yl)phenoxy(tert-butyl)dimethylsilane (Intermediate 16-3) (1.00 g, 2.78 mmol, 1 equiv), benzyl N-[2-(trifluoro-lambda4-boranyl)ethyl]carbamate potassium (952 mg, 3.34 mmol, 1.2 equiv), cesium carbonate (1.81 mg, 5.57 mmol, 2 equiv), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (204 mg, 0.278 mmol, 0.1 equiv) and RuPhos (130 mg, 0.278 mmol, 0.1 equiv) in toluene (10 mL) and water (1 mL) was stirred overnight at 80° C. under nitrogen atmosphere. The mixture was cooled to room temperature, diluted with ethyl acetate (100 mL), washed with water (50 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using a 0-100% gradient of ethyl acetate in petroleum ether to afford benzyl (3-((tert-butyldimethylsilyl)oxy)-4-(1,3-dioxolan-2-yl)phenethyl)carbamate (800 mg, 63%) as a colorless oil. MS (ESI) calculated for C25H35NO5Si: 457.23 m/z, found 458.25 [M+H]+.
To a solution of benzyl N-(2-{3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)phenyl}ethyl)carbamate (800 mg, 1.75 mmol, 1 equiv) in methanol (40 mL) and ethyl acetate (40 mL) was added 10% palladium on carbon (400 mg, 0.20 equiv) under nitrogen atmosphere. The mixture was stirred for 1 h at room temperature under hydrogen atmosphere. The resulting mixture was filtered through celite, and the filtrate was concentrated in vacuo to afford 2-(3-((tert-butyldimethylsilyl)oxy)-4-(1,3-dioxolan-2-yl)phenyl)ethan-1-amine (Intermediate 106-1) (1.2 g, 85%) as a yellow oil. MS (ESI) calculated for C17H29NO3Si: 323.19 m/z, found 324.20 [M+H]+.
Example 107 was prepared in a manner analogous to Example 19 using Intermediate 3-1 in place of Intermediate 19-2 and Intermediate 16-4 in place of Intermediate 19-3. MS (ESI) calculated for C31H31N7O3: 549.25 m/z, found 550.15 [M+H]+. 1H-NMR (300 MHz, DMSO-d6) δ (ppm): 10.20 (s, 1H), 7.95-8.03 (m, 2H), 7.60-7.69 (m, 3H), 7.46-7.49 (m, 2H), 7.12-7.14 (m, 1H), 6.89-6.94 (m, 3H), 6.36-6.40 (m, 1H), 4.26-4.27 (m, 2H), 3.61-3.68 (m, 4H), 3.31-3.41 (m, 4H), 3.20-3.25 (m, 2H), 2.94-2.99 (m, 2H).
Example 108 was prepared in a manner analogous to Example 16 using Intermediate 12-1 in place of Intermediate 16-2. MS (ESI) calculated for C37H35N7O3: 625.28 m/z, found 626.30 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.19 (s, 1H), 8.30-8.33 (m, 1H), 8.03-8.07 (m, 2H), 7.58-7.69 (m, 7H), 7.46-7.49 (m, 1H), 7.30-7.35 (m, 1H), 6.90-7.03 (m, 3H), 6.64-6.69 (m, 1H), 4.30 (s, 2H), 3.73-3.79 (m, 4H), 3.26-3.28 (m, 2H), 3.16-3.23 (m, 4H), 2.96-3.01 (m, 2H).
Example 109 was prepared in a manner analogous to Example 42 (via Intermediate 42-1) starting from N-methyl-N-(piperidin-4-yl)acetamide in place of 1-(piperidin-4-yl)pyrrolidin-2-one and using 1,3-dibromobenzene in place of 1,3-dibromo-2-fluorobenzene. MS (ESI) calculated for C41H42N8O3: 694.34 m/z, found 695.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.21 (s, 1H), 8.27-8.30 (s, 1H), 8.00-8.04 (m, 2H), 7.60-7.69 (m, 6H), 7.26-7.45 (m, 3H), 7.01-7.12 (m, 1H), 6.88-6.90 (m, 2H), 6.52-6.54 (m, 1H), 4.42-4.44 (m, 1H), 4.30 (s, 2H), 3.83-3.87 (m, 2H), 3.21-3.26 (m, 2H), 2.95-3.00 (m, 2H), 2.82 (s, 3H), 2.74 (s, 2H), 2.09 (s, 1H), 2.00 (s, 2H), 1.75-1.87 (m, 3H), 1.56-1.60 (m, 1H).
Example 110 was prepared in a manner analogous to Example 19 using Intermediate 4-1 in place of Intermediate 19-3 and Intermediate 98-3 in place of Intermediate 19-3. MS (ESI) calculated for C35H31N7O2: 581.25, found 582.15 [M+H]+. 1H-NMR (300 MHz, DMSO-d6) δ (ppm): 9.92 (s, 1H), 8.22-8.29 (m, 1H), 8.09 (s, 1H), 7.98-8.03 (m, 4H), 7.78-7.80 (m, 1H), 7.37-7.54 (m, 8H), 7.18-7.20 (m, 2H), 6.34-6.38 (m, 1H), 3.89 (s, 2H), 2.82-2.86 (m, 4H), 2.10-2.14 (m, 3H).
Triethylamine trihydrofluoride (45 mg, 0.28 mmol) was added to a solution of N-(2-{4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]phenyl}ethyl)-3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)benzamide (Intermediate 111-2) (90 mg, 0.14 mmol) in tetrahydrofuran (1 mL) and the mixture was stirred at room temperature for 30 min. The solvent was evaporated under reduced pressure and the crude product was purified by preparative HPLC on a XSelect CSH C18 OBD Column using a 27-47% gradient of acetonitrile in water to afford N-(2-{4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]phenyl}ethyl)-4-formyl-3-hydroxybenzamide (Example 111) (26 mg, 33%) as a yellow solid. MS (ESI) calculated for C33H26N6O3: 554.21 m/z, found 555.15 [M+H]+. 1HNMR (400 MHz, DMSO-d6) δ (ppm): 10.30 (s, 1H), 8.31-8.33 (m, 1H), 8.00-8.04 (m, 4H), 7.71-7.73 (m, 1H), 7.57-7.59 (m, 1H), 7.33-7.49 (m, 8H), 7.31-7.32 (m, 1H), 6.65-6.68 (m, 1H), 3.55-3.60 (m, 2H), 2.95-2.98 (m, 2H).
Intermediate 111-1 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 36-1 in place of the amine starting material and Intermediate 111-2 in place of Intermediate 1-4. MS (ESI) calculated for C39H40N6O3Si: 668.29 m/z, found 669.00 [M+H]+.
Intermediate 111-2 was prepared in a manner analogous to Intermediate 3-4 using Intermediate 16-3 in place of Intermediate 3-3. MS (ESI) calculated for C18H21ClN4O4: 392.13 m/z, found 393.15 [M+H]+.
Example 112 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 106-1 in place of the amine starting material, Intermediate 112-1 in place of Intermediate 1-4, HATU in place of PyBOP and tetrahydrofuran/triethylamine trihydrofluoride (10:1) in place of 2,2,2-trifluoroacetic acid/methanesulfonic acid. MS (ESI) calculated for C32H25N7O3: 555.20 m/z, found 556.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.10 (s, 1H), 8.71-8.76 (m, 1H), 8.47-8.60 (m, 3H), 8.17-8.25 (m, 2H), 8.11-8.17 (m, 2H), 7.93-8.00 (m, 1H), 7.63-7.69 (m, 1H), 7.54-7.62 (m, 2H), 7.46-7.53 (m, 1H), 7.23-7.30 (m, 1H), 6.94-7.00 (m, 1H), 6.89-6.93 (m, 1H), 3.51-3.61 (m, 2H), 2.87-2.95 (m, 2H).
A solution of methyl 6-aminonicotinate (1.52 g, 1.00 mmol, 1 equiv), 2-chloro-3-nitro-6-phenylpyridine (2.34 g, 1.00 mmol, 1 equiv), tris(dibenzylideneacetone)dipalladium(0) (46 mg, 0.050 mmol, 0.05 equiv), RuPhos (47 mg, 0.1 mmol, 0.1 equiv), and sodium carbonate (2.08 g, 2.00 mmol, 2.0 equiv) in 1,4-dioxane (100 mL) was stirred at 100° C. overnight under nitrogen atmosphere. The mixture was cooled to room temperature and quenched with water (30 mL). The resulting mixture was extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue obtained was purified by silica gel chromatography using a 0-50% gradient of ethyl acetate in petroleum ether to afford methyl 6-((3-nitro-6-phenylpyridin-2-yl)amino)nicotinate as a yellow solid (1.8 g, 51%). MS (ESI) calculated for C18H14N4O4: 350.10 m/z, found 351.10 [M+H]+.
A solution of methyl 6-((3-nitro-6-phenylpyridin-2-yl)amino)nicotinate (1.8 g, 5.1 mmol, 1 equiv), 2-aminonicotinaldehyde (0.752 g, 6.17 mmol, 1.20 equiv) and sodium dithionite (2.68 g, 15.4 mmol, 3.0 equiv) in dimethyl sulfoxide (15 mL) and methanol (3 mL) was refluxed at 100° C. overnight. The resulting mixture was cooled to room temperature and quenched with water (50 mL). The mixture was extracted with ethyl acetate (3×100 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue obtained was purified by silica gel chromatography using a 0-80% gradient of ethyl acetate in petroleum ether to afford methyl 6-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl)nicotinate as a yellow solid (1.5 g, 69%). MS (ESI) calculated for C24H18N6O2: 422.15 m/z, found 423.10 [M+H]+.
To a solution of methyl 6-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl)nicotinate (1.5.0 g, 3.55 mmol, 1 equiv) in tetrahydrofuran (30 mL) and water (30 mL) was added lithium hydroxide (5.0 mL, 2 M in water). The resulting mixture was stirred at room temperature for 1 h. The mixture was concentrated and purified by reverse-phase flash column chromatography on C18 silica gel to afford 6-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl)nicotinic acid (Intermediate 112-1) (1.0 g, 69%) as a yellow solid. MS (ESI) calculated for C23H16N6O2: 408.13 m/z, found 409.10 [M+H]+.
Example 113 was prepared in a manner analogous to Example 19 using Intermediate 113-1 in place of Intermediate 19-3 and Intermediate 67-1 in place of Intermediate 19-2. MS (ESI) calculated for C32H27N7O2: 541.61 m/z, found 542.25 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.19 (s, 1H), 9.26 (s, 1H), 8.65-8.66 (m, 1H), 8.50-8.52 (m, 1H), 8.40-8.42 (m, 1H), 8.14-8.16 (m, 1H), 8.07-8.08 (m, 1H), 7.65-7.68 (m, 7H), 6.89-6.90 (m, 2H), 6.69-6.70 (m, 1H), 4.29 (s, 2H), 3.23-3.25 (m, 2H), 2.95-2.97 (m, 2H).
Intermediate 113-1 was prepared in a manner analogous to Intermediate 16-4 using Intermediate 1-3 in place of Intermediate 16-3. MS (ESI) calculated for C19H20O5: 328.13 m/z, found 329.15 [M+H]+.
Example 114 was prepared in a manner analogous to Example 19 using Intermediate 74-1 in place of Intermediate 19-2 and Intermediate 113-1 in place of Intermediate 19-3. MS (ESI) calculated for C27H24N6O2: 464.20 m/z, found 465.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.21 (s, 1H), 8.39-8.40 (m, 1H), 8.27-8.29 (m, 1H), 8.05-8.07 (m, 1H), 7.56-7.69 (m, 6H), 7.44-7.48 (m, 1H), 6.89-6.91 (m, 2H), 6.62-6.64 (m, 1H), 4.29 (s, 2H), 3.22-3.27 (m, 2H), 2.95-3.00 (m, 2H).
Example 115 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 115-1 in place of the amine starting material and Intermediate 28-2 in place of Intermediate 1-4. MS (ESI) calculated for C34H28N6O3: 568.22 m/z, found 569.20 [M+H]+. 1H-NMR (300 MHz, DMSO-d6) δ (ppm): 10.21 (s, 1H), 8.71 (s, 1H), 8.02-8.25 (m, 4H), 7.44-7.58 (m, 9H), 7.20-7.27 (m, 1H), 6.92-6.95 (m, 1H), 6.37-6.39 (m, 1H), 5.01 (s, 1H), 3.34 (s, 2H), 1.53 (s, 3H).
Intermediate 115-1 was prepared in a manner analogous to Intermediate 11-1 using tert-butyl (1-(4-aminophenyl)ethyl)carbamate in place of tert-butyl 2-(4-aminophenyl)acetate. MS (ESI) calculated for C25H22N6: 406.19 m/z, found: 407.20 [M+H]+.
Example 116 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 115-1 in place of the amine starting material. MS (ESI) calculated for C34H28N6O3: 568.22 m/z, found 569.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.61-10.75 (m, 1H), 10.08-10.31 (m, 1H), 8.64-8.73 (m, 1H), 8.23-8.37 (m, 1H), 7.93-8.09 (m, 4H), 7.73-7.90 (m, 1H), 7.33-7.65 (m, 10H), 6.81-6.96 (m, 2H), 6.60-6.71 (m, 1H), 4.95-5.11 (m, 1H), 3.40-3.57 (m, 2H), 1.34-1.51 (m, 3H).
Example 117 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material and Intermediate 100-2 in place of Intermediate 1-4. MS (ESI) calculated for C34H28N6O3: 568.22 m/z, found 569.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.03 (s, 1H), 8.34-8.36 (m, 1H), 8.01-8.06 (m, 4H), 7.75-7.77 (m, 1H), 7.59-7.61 (m, 1H), 7.47-7.56 (m, 3H), 7.38-7.40 (m, 2H), 7.21-7.23 (m, 2H), 6.84-6.91 (m, 2H), 6.80-6.82 (m, 1H), 4.34-4.41 (m, 2H), 2.89-2.93 (m, 2H), 2.49-2.50 (m, 2H).
Example 118 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 28-1 in place of the amine starting material, Intermediate 100-2 in place of Intermediate 1-4 and T3P in place of PyBOP. MS (ESI) calculated for C33H26N6O3: 554.21 m/z, found 555.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.71 (s, 1H), 10.14-10.29 (m, 2H), 8.26-8.35 (m, 1H), 8.01-8.11 (m, 4H), 7.73-7.89 (m, 4H), 7.60-7.65 (m, 2H), 7.40-7.54 (m, 5H), 6.85-6.95 (m, 2H), 6.68-6.75 (m, 1H), 2.90-3.00 (m, 2H), 2.63-2.76 (m, 2H).
Example 119 was prepared in a manner analogous to Example 19 using Intermediate 119-1 in place of Intermediate 19-2 and Intermediate 113-1 in place of Intermediate 19-3. MS (ESI) calculated for C32H27N7O2: 541.22 m/z, found 542.20 [M+H]+. 1H NMR (300 MHz, methanol-d4) δ (ppm): 8.74 (s, 2H), 8.42-8.46 (m, 3H), 8.32-8.35 (m, 1H), 8.01-8.03 (m, 1H), 7.76-7.78 (m, 2H), 7.67-7.73 (m, 3H), 7.30-7.33 (m, 1H), 6.66-6.79 (m, 3H), 5.57 (s, 1H), 4.38 (s, 2H), 3.35-3.37 (m, 2H), 2.95-3.03 (m, 2H). Note that the aldehyde proton is not visible for some compounds when dissolved in methanol-d4.
Intermediate 119-1 was prepared in a manner analogous to Intermediate 12-1 using pyridin-4-ylboronic acid in place of (3-morpholinophenyl)boronic acid, [1,1′-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II)/tribasic potassium phosphate in place of tetrakis(triphenylphosphine)palladium(0)/sodium bicarbonate and 1,4-dioxane in place of toluene/ethanol. MS (ESI) calculated for C23H19N7: 393.17 m/z, found 394.20 [M+H]+.
Example 120 was prepared in a manner analogous to Example 19 using Intermediate 4-1 in place of Intermediate 19-2 and Intermediate 120-1 in place of Intermediate 19-3. MS (ESI) calculated for C33H27ClN6O2: 574.19 m/z, found 575.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 11.09-11.46 (m, 1H), 10.14 (s, 1H), 9.05-9.32 (m, 2H), 8.24-8.42 (m, 1H), 7.95-8.17 (m, 4H), 7.63-7.82 (m, 5H), 7.36-7.60 (m, 5H), 7.03-7.20 (m, 1H), 6.44-6.64 (m, 1H), 4.31-4.44 (m, 2H), 3.15-3.37 (m, 4H).
Intermediate 120-1 was prepared in a manner analogous to Intermediate 16-4 using Intermediate 13-1 in place of Intermediate 16-3. MS (ESI) calculated for C19H19ClO5: 362.09 m/z, found 361.05 [M−H]−.
Example 121 was prepared in a manner analogous to Example 19 using Intermediate 102-1 in place of Intermediate 19-2 and Intermediate 113-1 in place of Intermediate 19-3. MS (ESI) calculated for C35H31N7O3: 597.25 m/z, found 598.25 [M+H]+. 1H NMR (300 MHz, methanol-d4) δ (ppm): 8.40 (s, 1H), 8.26-8.29 (m, 1H), 7.95-7.98 (m, 2H), 7.65-7.75 (m, 6H), 7.40-7.48 (m, 1H), 7.35-7.39 (m, 1H), 7.29-7.32 (m, 1H), 6.74-6.81 (m, 2H), 6.62-6.69 (m, 1H), 5.57 (s, 1H), 4.36 (s, 2H), 3.34-3.40 (m, 2H), 2.91-3.05 (m, 2H), 2.14 (s, 3H). Note that the aldehyde signal is not visible for some compounds dissolved in methanol-d4.
Example 122 was prepared in a manner analogous to Example 19 using Intermediate 4-1 in place of Intermediate 19-2 and Intermediate 88-1 in place of Intermediate 19-3. MS (ESI) calculated for C33H27FN6O2: 558.22 m/z, found 559.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.98-11.02 (s, 1H), 10.25 (s, 1H), 9.10-9.12 (m, 2H), 8.31-8.34 (m, 1H), 7.95-8.05 (m, 4H), 7.60-7.70 (m, 4H), 7.41-7.50 (m, 6H), 6.90-6.95 (m, 1H), 6.54 (s, 1H), 4.32-4.37 (m, 2H), 3.24 (s, 2H), 3.00-3.09 (m, 2H).
Example 123 was prepared in a manner analogous to Example 19 using Intermediate 4-1 in place of Intermediate 19-2 and Intermediate 123-2 in place of Intermediate 19-3. MS (ESI) calculated for C33H27FN6O2: 558.22 m/z, found 559.23 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm) 11.32 (s, 1H), 10.24 (s, 1H), 9.04 (s, 2H), 8.33 (d, J=8.4 Hz, 1H), 8.05-8.00 (m, 4H), 7.73-7.60 (m, 4H), 7.53-7.36 (m, 5H), 6.78 (d, J=10.3 Hz, 2H), 6.56 (dd, J=7.6, 5.3 Hz, 1H), 4.30 (s, 2H), 3.29-3.25 (m, 2H), 2.98 (t, J=7.9 Hz, 2H).
Intermediate 123-2 was prepared in a manner analogous to Intermediate 16-4 using Intermediate 123-1 in place of Intermediate 16-3. MS (ESI) calculated for C19H19FO5: 346.12 m/z, found 347.14 [M+H]+.
Intermediate 123-1 was prepared in a manner analogous to Intermediate 1-3 (via Intermediate 1-2) starting from 4-bromo-2-fluoro-6-hydroxybenzaldehyde in place of 4-bromo-2-hydroxybenzaldehyde. MS (ESI) calculated for C17H16BrFO4: 382.02 m/z, found 383.00 [M+H]+.
Example 124 was prepared in a manner analogous to Intermediate 19 using Intermediate 124-2 in place of Intermediate 19-2 and Intermediate 113-1 in place of Intermediate 19-3. MS (ESI) calculated for C33H28N6O2: 540.62 m/z, found 541.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.20 (s, 1H), 8.69-8.70 (m, 1H), 8.52-8.53 (m, 1H), 8.05-8.07 (m, 1H), 7.79-7.81 (m, 2H), 7.62-7.69 (m, 3H), 7.60-7.62 (m, 3H), 7.55-7.57 (m, 2H), 7.43-7.53 (m, 1H), 6.91-6.93 (m, 2H), 6.64-6.68 (m, 1H), 4.30 (s, 2H), 3.24-3.29 (m, 2H), 2.97-3.00 (m, 2H).
Intermediate 124-2 was prepared in a manner analogous to Intermediate 12-1 using Intermediate 124-1 in place of Intermediate 1-1 and phenyl boronic acid in place of (3-morpholinophenyl)boronic acid. MS (ESI) calculated for C24H20N6: 392.17 m/z, found 393.15 [M+H]+.
Intermediate 124-1 was prepared in a manner analogous to Intermediate 1-1 using 2,5-dichloro-3-nitropyridine in place of 2,6-dichloro-3-nitropyridine. MS (ESI) calculated for C23H23ClN6O2: 450.16 m/z, found 451.15 [M+H]+.
Example 125 was prepared in a manner analogous to Example 19 using Intermediate 4-1 in place of Intermediate 19-2 and Intermediate 125-2 in place of Intermediate 19-3. MS (ESI) calculated for C33H28N6O: 524.23 m/z, found 525.20 [M+H]+. 1H-NMR (400 MHz, DMSO-d6) δ (ppm): 9.95 (s, 1H), 8.21-8.28 (m, 1H), 7.84-8.04 (m, 4H), 7.79-7.83 (m, 2H), 7.28-7.57 (m, 10H), 7.17-7.19 (m, 1H), 7.01-7.15 (m, 2H), 6.33-6.37 (m, 1H), 3.86 (s, 2H), 2.68-2.88 (m, 4H).
Intermediate 125-2 was prepared in a manner analogous to Intermediate 16-4 using Intermediate 125-1 in place of Intermediate 16-3. MS (ESI) calculated for C11H12O3: 192.08 m/z, found 193.15 [M+H]+.
Intermediate 125-1 was prepared in a manner analogous to Intermediate 1-2 using 4-bromobenzaldehyde in place of 4-bromo-2-hydroxybenzaldehyde. MS (ESI) calculated for C9H9BrO2: 227.98 m/z, found 229.00 [M+H]+.
Example 126 was prepared in a manner analogous to Example 19 using Intermediate 115-1 in place of Intermediate 19-2 and Intermediate 92-3 in place of Intermediate 19-3. MS (ESI) calculated for C34H30N6O2, 554.24 m/z: found 555.25 [M+H]+. 1H NMR (300 MHz, methanol-d4) δ (ppm) 10.02 (s, 1H), 8.53 (s, 1H), 8.23 (d, J=8.4 Hz, 1H), 8.08-8.00 (m, 2H), 8.00-7.94 (m, 2H), 7.68-7.52 (m, 4H), 7.48-7.30 (m, 5H), 6.93 (d, J=8.5 Hz, 1H), 6.41 (dd, J=7.7, 5.0 Hz, 1H), 4.30 (d, J=6.7 Hz, 1H), 3.09-2.99 (m, 1H), 2.80-2.99 (m, 3H), 1.65 (d, J=6.7 Hz, 3H).
Example 127 was prepared in a manner analogous to Example 19 using Intermediate 115-1 in place of Intermediate 19-2 and Intermediate 113-1 in place of Intermediate 19-3. MS (ESI) calculated for C36H30N6O2: 580.26 m/z, found 581.25 [M+H]+. 1H NMR (300 MHz, methanol-d4) δ (ppm) 8.30 (d, J=8.4 Hz, 1H), 8.09-7.93 (m, 4H), 7.65-7.75 (m, 6H), 7.48-7.38 (m, 3H), 7.32 (d, J=7.8 Hz, 1H), 6.72 (d, J=8.8 Hz, 1H), 6.58-6.50 (m, 1H), 5.59 (s, 1H), 4.61 (d, J=6.8 Hz, 1H), 3.13-2.87 (m, 4H), 1.81 (d, J=6.8 Hz, 3H). Note that the aldehyde signal is not visible for some compounds dissolved in methanol-d4.
Example 128 was prepared in a manner analogous to Example 19 using Intermediate 4-1 in place of Intermediate 19-2 and Intermediate 128-3 in place of Intermediate 19-3. MS (ESI) calculated for C33H27FN6O2: 558.22 m/z, found 559.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm) 10.87 (br s, 1H), 10.23 (d, J=2.7 Hz, 1H), 9.29-9.06 (m, 2H), 8.34 (d, J=8.4 Hz, 1H), 8.12-8.00 (m, 4H), 7.73-7.60 (m, 41H), 7.57-7.34 (m, 6H), 6.98 (d, J=6.2 Hz, 1H), 6.66-6.55 (in 1H), 4.37-4.27 (m, 2H), 3.31-3.17 (m, 2H), 3.07-2.94 (m, 2H).
Intermediate 128-3 was prepared in a manner analogous to Intermediate 16-4 using Intermediate 128-2 in place of Intermediate 16-3. MS (ESI) calculated for C19H19FO5: 346.12 m/z, found 369.00 [M+Na]+.
Intermediate 128-2 was prepared in a manner analogous to Intermediate 1-3 (via Intermediate 1-2) starting from Intermediate 128-1 in place of 4-bromo-2-hydroxybenzaldehyde. 1H NMR (300 MHz, chloroform-d) 6 (ppm) 7.41-7.25 (m, 3H), 7.13 (d, J=5.5 Hz, 1H), 7.01-6.86 (m, 2H), 6.11 (d, J=1.4 Hz, 1H), 5.03 (s, 2H), 4.21-3.95 (m, 4H), 3.84 (s, 3H).
To a cooled (0° C.) solution of methyl 4-bromo-5-fluoro-2-hydroxybenzoate (2.00 g, 8.03 mmol, 1 equiv) in tetrahydrofuran (10 mL) was added borane-tetrahydrofuran complex (1M in tetrahydrofuran, 40 mL, 40 mmol, 5 equiv) and the resulting mixture was stirred at room temperature overnight. The reaction was quenched with ice/water (50 mL) and the resulting mixture was extracted with ethyl acetate (3×80 mL). The organic layers were combined, dried over sodium sulfate, filtered, and concentrated in vacuo to give 5-bromo-4-fluoro-2-(hydroxymethyl) phenol (2 g, crude quant.) as a yellow oil, which was used without further purification in the next step. MS (ESI) calculated for C7H6BrFO2: 219.95 m/z, found 243.90 [M+Na]+.
To a solution of 5-bromo-4-fluoro-2-(hydroxymethyl) phenol (2.00 g, 9.05 mmol, 1 equiv) in dichloromethane (40 mL) was added manganese (IV) oxide (6.29 g, 72.4 mmol, 8 equiv). The reaction mixture was stirred overnight at 40° C. The resulting mixture was cooled to room temperature and concentrated in vacuo. The resulting crude material was purified by silica gel column chromatography using a 0-60% gradient of ethyl acetate in petroleum ether to give 4-bromo-5-fluoro-2-hydroxybenzaldehyde (Intermediate 128-1) (1 g, 50%) as an off-white solid. 1H NMR (300 MHz, chloroform-d) 6 (ppm) 10.86 (s, 1H), 9.84 (d, J=0.6 Hz, 1H), 7.32 (d, J=7.4 Hz, 1H), 7.30-7.25 (m, 1H).
Example 129 was prepared in a manner analogous to Example 19 using Intermediate 4-1 in place of Intermediate 19-2 and Intermediate 129-1 in place of Intermediate 19-3. MS (ESI) calculated for C34H30N6O2: 554.24 m/z, found 555.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm) 10.24 (s, 1H), 8.28 (d, J=8.4 Hz, 1H), 8.08-7.95 (m, 4H), 7.59-7.53 (m, 2H), 7.51-7.36 (m, 6H), 7.18 (d, J=7.4 Hz, 1H), 6.96 (d, J=22.5 Hz, 2H), 6.37 (s, 1H), 3.91 (s, 2H), 2.62-2.63 (s, 2H), 2.50-2.46 (m, 2H), 1.79-1.80 (m, 2H).
A mixture of 2-{5-bromo-2-[(4-methoxyphenyl)methoxy]phenyl}-1,3-dioxolane (Intermediate 3-3) (2.00 g, 5.48 mmol, 1 equiv), tributyl(prop-2-en-1-yl)stannane (3.63 g, 11.0 mmol, 2 equiv) and bis(triphenylphosphine)palladium(II) dichloride (0.38 g, 0.55 mmol, 0.1 equiv) in N,N-dimethylformamide (25 mL) was stirred at 80° C. for 1 h under nitrogen atmosphere. The reaction was quenched with water (50 mL) and extracted with ethyl acetate (3×50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by silica gel column chromatography using a 0-75% gradient of ethyl acetate in petroleum ether to afford 2-{2-[(4-methoxyphenyl)methoxy]-5-(prop-2-en-1-yl)phenyl}-1,3-dioxolane (1.5 g, 84%) as a yellow solid. MS (ESI) calculated for C20H22O4: 326.15 m/z, found 327.20 [M+H]+.
To a cooled (0° C.) solution of 2-{2-[(4-methoxyphenyl)methoxy]-5-(prop-2-en-1-yl)phenyl}-1,3-dioxolane (600 mg, 1.84 mmol, 1 equiv) in tetrahydrofuran (25 mL) was added BH3·tetrahydrofuran (1M in tetrahydrofuran, 2 mL, 2.0 mmol, 1.1 equiv), sodium hydroxide (368 mg, 9.19 mmol, 5 equiv) and hydrogen peroxide (313 mg, 9.19 mmol, 5 equiv). The resulting mixture was stirred at room temperature for 12 h. The reaction was quenched with water (50 mL) and the mixture was extracted with ethyl acetate (3×50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by silica gel column chromatography using a 0-54% gradient of ethyl acetate in petroleum ether to afford 3-[3-(1,3-dioxolan-2-yl)-4-[(4-methoxyphenyl)methoxy]phenyl]propan-1-ol (360 mg, 57%) as a light-yellow oil. MS (ESI) calculated for C20H24O5: 344.16 m/z, found 367.05 [M+Na]+.
To a mixture of oxalyl chloride (193 mg, 1.52 mmol, 1.5 equiv) in dichloromethane (6 mL) at −78° C. was added dimethyl sulfoxide (238 mg, 3.05 mmol, 3 equiv) dropwise over 10 min under nitrogen atmosphere and the resulting mixture was stirred at −78° C. for 0.5 h. A solution of 3-[3-(1,3-dioxolan-2-yl)-4-[(4-methoxyphenyl)methoxy]phenyl]propan-1-ol (350 mg, 1.02 mmol, 1 equiv) in dichloromethane was added dropwise. The mixture was stirred at −78° C. for additional 0.5 h. Triethylamine (514 mg, 5.08 mmol, 5 equiv) was added. The mixture was then allowed to warm to room temperature and the reaction was quenched by the addition of water (20 mL). The resulting mixture was extracted with ethyl acetate (3×50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The obtained residue was purified by silica gel column chromatography using a 0-42% gradient of ethyl acetate in petroleum ether to afford 3-[3-(1,3-dioxolan-2-yl)-4-[(4-methoxyphenyl)methoxy]phenyl]propanal (Intermediate 129-1) (180 mg, 52%) as a white solid. MS (ESI) calculated for C20H22O5: 342.15 m/z, found 365.05 [M+Na]+.
Example 130 was prepared in a manner analogous to Example 19 using Intermediate 4-1 in place of Intermediate 19-2 and Intermediate 130-1 in place of Intermediate 19-3. MS (ESI): mass calculated for C34H30N6O2: 554.24 m/z, found 555.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm) 10.81 (s, 1H), 10.19 (s, 1H), 8.96 (s, 2H), 8.34 (d, J=8.4 Hz, 1H), 8.13-8.00 (m, 4H), 7.74-7.57 (m, 5H), 7.56-7.39 (m, 5H), 6.86 (d, J=6.6 Hz, 2H), 6.64 (dd, J=7.6, 5.5 Hz, 1H), 4.28 (d, J=6.3 Hz, 2H), 2.69 (t, J=7.6 Hz, 2H), 2.62-2.56 (m, 2H), 2.05-1.89 (m, 2H).
Intermediate 130-1 was prepared in a manner analogous to Intermediate 129-1 starting from Intermediate 1-3 in place of Intermediate 3-3. MS (ESI): mass calculated for C20H22O5 342.15 m/z, found 365.10 [M+Na]+.
Example 131 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 131-2 in place of the amine component, Intermediate 29-2 in place of Intermediate 1-4 and tetrahydrofuran/triethylamine trihydrofluoride in place of 2,2,2-trifluoroacetic acid/methanesulfonic acid. MS (ESI) calculated for C33H26N6O3: 554.21 m/z, found 555.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm) 10.71 (s, 1H), 10.20 (s, 1H), 8.77 (t, J=5.6 Hz, 1H), 8.35 (d, J=8.4 Hz, 2H), 8.13-8.02 (m, 4H), 8.08-7.93 (m, 2H), 7.77-7.57 (m, 4H), 7.61-7.37 (m, 4H), 6.97-6.81 (m, 2H), 6.75 (dd, J=7.6, 5.7 Hz, 1H), 3.61-3.48 (m, 1H), 2.88 (t, J=7.2 Hz, 2H), 2.21 (s, 1H).
Intermediate 131-2 was prepared in a manner analogous to Intermediate 106-1 using Intermediate 131-1 in place of Intermediate 16-3. MS (ESI) calculated for C17H29NO3Si: 323.19 m/z, found 324.20 [M+H]+.
Intermediate 131-1 was prepared in a manner analogous to Intermediate 16-3 starting from Intermediate 3-2 in place of Intermediate 1-2. MS (ESI) calculated for C15H23BrO3Si: 358.06 m/z, found 359.05 [M+H]+.
Example 132 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 4-1 in place of the amine starting material. MS (ESI) calculated for C33H26N6O3: 554.21 m/z, found 555.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.17 (s, 1H), 8.71-8.74 (m, 1H), 8.24-8.27 (m, 1H), 7.99-8.02 (m, 4H), 7.59-7.61 (m, 1H), 7.40-7.60 (m, 7H), 7.19-7.20 (m, 1H), 6.91 (s, 1H), 6.85-6.89 (m, 1H), 6.39-6.42 (m, 1H), 4.38 (s, 2H), 3.56 (s, 2H).
Example 133 was prepared in a manner analogous to Example 16 using Intermediate 4-1 in place of Intermediate 16-2. MS (ESI) calculated for C33H28N6O2: 540.23 m/z, found 541.25 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.19 (s, 1H), 9.10 (s, 1H), 8.25-8.30 (m, 1H), 8.00-8.10 (m, 4H), 7.20-7.70 (m, 9H), 6.85-6.93 (m, 2H), 6.39-6.43 (m, 1H), 4.00-4.16 (m, 2H), 2.90-3.15 (m, 4H).
To a mixture of 3-{3-[4-(1-aminocyclobutyl)phenyl]-5-phenylimidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (80 mg, 0.19 mmol, 1 equiv) and 2-{3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)phenyl}acetaldehyde (Intermediate 113-1) (60 mg, 0.19 mmol, 1.0 equiv) in methanol (3 mL) was added sodium cyanoborohydride (58 mg, 0.93 mmol, 5.0 equiv). The mixture was stirred at room temperature for 1 h under nitrogen atmosphere. The reaction was quenched with water (10 mL) and extracted with ethyl acetate (3×10 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography using a 0-20% gradient of methanol in dichloromethane to provide 3-(3-{4-[1-({2-[4-(1,3-dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]ethyl}amino)cyclobutyl]phenyl}-5-phenylimidazo [4,5-b]pyridin-2-yl)pyridin-2-amine (30 mg, 22%) as a yellow solid. MS (ESI) calculated for C46H44N6O4, 744.34 m/z, found 745.30 [M+H]+.
To a solution of 3-(3-{4-[1-({2-[4-(1,3-Dioxolan-2-yl)-3-[(4-methoxyphenyl)methoxy]phenyl]ethyl}amino)cyclobutyl]phenyl}-5-phenylimidazo[4,5-b]pyridin-2-yl)pyridin-2-amine (30 mg, 0.040 mmol, 1 equiv) in dichloromethane (2.8 mL) was added 2,2,2-trifluoroacetic acid (0.2 mL) and the resulting mixture was stirred at room temperature for 30 min. The resulting mixture was concentrated in vacuo and the resulting oil was purified by preparative HPLC on a XSelect CSH C18 OBD Column using a 10-24% gradient of acetonitrile in water (+0.05% 2,2,2-trifluoroacetic acid) to afford 4-(2-((1-(4-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl)phenyl)cyclobutyl)amino)ethyl)-2-hydroxybenzaldehyde (Example 134) (7.0 mg, 25%) as a yellow solid. MS (ESI) calculated for C36H32N6O2: 580.26 m/z, found 581.30 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm) 10.00 (s, 1H), 8.30 (d, J=8.4 Hz, 1H), 7.98 (d, J=8.4 Hz, 1H), 7.95-7.89 (m, 2H), 7.86 (dd, J==6.0, 1.7 Hz, 2H), 7.76-7.69 (m, 2H), 7.65-7.53 (m, 3H), 7.41-7.32 (m, 3H), 6.80-6.69 (m, 2H), 6.58-6.49 (m, 1H), 2.82-2.62 (m, 8H), 2.18-2.03 (m, 1H), 1.86-1.71 (m, 1H).
A mixture of 3-(3-(4-(aminomethyl) phenyl)-5-phenyl-3H-imidazo[4,5-b]pyridin-2-yl) pyridin-2-amine (Intermediate 4-1) (200 mg, 0.51 mmol, 1 equiv), 2-{4-[(tert-butyldimethylsilyl) oxy]-3-(1,3-dioxolan-2-yl) phenyl}acetaldehyde (Intermediate 135-1) (197 mg, 0.612 mmol, 1.2 equiv) and sodium cyanoborohydride (640 mg, 10.2 mmol, 20 equiv) in methanol (10 mL) was stirred at room temperature for 2 h. The reaction was quenched with water (20 mL) and the resulting mixture was extracted with ethyl acetate (20 mL×3). The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by flash column chromatography on silica gel using a 0-100% gradient of ethyl acetate in petroleum ether to afford 3-(3-(4-(((4-((tert-butyldimethylsilyl) oxy)-3-(1,3-dioxolan-2-yl) phenethyl) amino) methyl) phenyl)-5-phenyl-3H-imidazo[4,5-b]pyridin-2-yl) pyridin-2-amine (192 mg, 54%) as a white oil. MS (ESI) calculated for C41H46N6O3Si: 698.94 m/z, found 699.15 [M+H]+
Triethylamine trihydrofluoride (1 mL) was added to a solution of 3-(3-(4-(((4-((tert-butyldimethylsilyl) oxy)-3-(1,3-dioxolan-2-yl) phenethyl) amino) methyl) phenyl)-5-phenyl-3H-imidazo[4,5-b]pyridin-2-yl) pyridin-2-amine (90 mg, 0.14 mmol, 1 equiv) in tetrahydrofuran (10 mL). The mixture was stirred at room temperature for 1 h. The mixture was concentrated in vacuo and the resulting residue was purified by preparative HPLC on a XSelect CSH Fluoro Phenyl column using a 8-38% gradient of acetonitrile in water (+0.05% 2,2,2-trifluoroacetic acid) to afford 5-(2-((4-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl) benzyl) amino) ethyl)-2-hydroxybenzaldehyde (Example 135) (5.9 mg, 5.94%) as an off-white solid. MS (ESI) calculated for C33H28N6O2: 540.23 m/z, found 541.25 [M+H]+. 1H NMR (300 MHz, methanol-d4) δ (ppm) 8.31 (d, J=8.5 Hz, 1H), 8.15-7.89 (m, 4H), 7.89-7.61 (m, 5H), 7.51-7.30 (m, 4H), 6.86-6.64 (m, 3H), 5.60 (s, 1H), 4.39 (s, 2H), 3.03 (s, 2H), 2.98 (d, J=7.9 Hz, 2H). Note that the aldehyde signal is not visible for some compounds dissolve in methanol-d4.
Intermediate 135-1 was prepared in a manner analogous to Intermediate 16-4 using Intermediate 131-1 in place of Intermediate 16-3. MS (ESI) calculated for C17H26O4Si: 322.16 m/z, found 323.15 [M+H]+.
A mixture of 2-chloro-3-nitro-6-phenylpyridine (10.0 g, 42.6 mmol, 1 equiv), methyl 4-aminobenzoate (7.73 g, 51.1 mmol, 1.2 equiv), tris(dibenzylideneacetone)dipalladium(0) (3.90 g, 4.26 mmol, 0.1 equiv), RuPhos (1.99 g, 4.26 mmol, 0.1 equiv) and sodium carbonate (9.03 g, 85.2 mmol, 2.0 equiv) in 1,4-dioxane (200 mL) was stirred for 4 h at 90° C. under nitrogen atmosphere. The mixture was cooled to room temperature and quenched with water (200 mL). The resulting mixture was extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by silica gel column chromatography using a 0-100% gradient of ethyl acetate in petroleum ether to afford methyl 4-[(3-nitro-6-phenylpyridin-2-yl) amino]benzoate (14.0 g, 81%) as a red/brown solid. MS (ESI) calculated for C19H15N3O4: 349.11 m/z, found 350.05 [M+H]+.
To a stirred mixture of methyl 4-[(3-nitro-6-phenylpyridin-2-yl) amino]benzoate (14.0 g, 40.1 mmol, 1.0 equiv) in ethanol (100 mL) and saturated aqueous ammonium chloride (100 mL) was added iron (11.19 g, 200.4 mol, 5.0 equiv) in portions under nitrogen atmosphere and the resulting mixture was stirred for 2 h at 70° C. The mixture was cooled to room temperature and extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by silica gel column chromatography using a 0-50% gradient of ethyl acetate in petroleum ether to provide methyl 4-[(3-amino-6-phenylpyridin-2-yl) amino]benzoate (11.0 g, 85%) as a yellow solid. MS (ESI) calculated for C19H17N3O2: 319.13 m/z, found 320.05 [M+H]+.
To a stirred solution of methyl 4-[(3-amino-6-phenylpyridin-2-yl) amino]benzoate (11 g, 34 mmol, 1.0 equiv) and 2-aminopyridine-3-carbaldehyde (4.63 g, 37.9 mmol, 1.1 equiv) in anhydrous ethanol (350 mL) was added sodium bisulfite (10.75 g, 103.3 mmol, 3 equiv). The resulting mixture was stirred overnight at 90° C. The resulting mixture was cooled to room temperature and quenched with water (300 mL). The precipitated solids were collected by filtration and washed with ethyl acetate (3×50 mL). The resulting product was dried to afford methyl 4-(2-(2-aminopyridin-3-yl)-5-phenyl-1,2-dihydro-3H-imidazo[4,5-b]pyridin-3-yl)benzoate (12.0 g, 82%) as an orange solid. MS (ESI) calculated for C25H21N5O2: 423.17 m/z, found 424.25 [M+H]+.
To a stirred solution of methyl 4-(2-(2-aminopyridin-3-yl)-5-phenyl-1,2-dihydro-3H-imidazo[4,5-b]pyridin-3-yl)benzoate (12.0 g, 28.3 mmol, 1.0 equiv) in anhydrous 1,4-dioxane was added (diacetoxyiodo)benzene (27.38 g, 85.01 mmol, 3 equiv) at room temperature and the resulting mixture was stirred at room temperature for 3 h. The reaction was quenched with water (200 mL) and the precipitated solids were collected by filtration and dried by air to afford methyl 4-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl) benzoate (11.0 g, 91%) as a yellow solid. MS (ESI) calculated for C25H19N5O2: 421.15 m/z, found 422.10 [M+H]+.
To a cooled (0° C.) solution of methyl 4-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl) benzoate (1.00 g, 2.37 mmol, 1 equiv) in tetrahydrofuran (70 mL) was added dropwise lithium aluminum hydride (2.5 M in tetrahydrofuran, 2.8 mL, 7.2 mmol) under nitrogen atmosphere. The mixture was stirred for 4 h at room temperature. The reaction was quenched with saturated aqueous ammonium chloride (40 mL) at 0° C. and the resulted mixture was extracted with chloroform/isopropanol (3:1, 2×40 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give (4-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl) phenyl) methanol (900 mg, 82%) as a brown solid. MS (ESI) calculated for C24H19N5O: 393.16 m/z, found 394.25 [M+H]+.
To a cooled (0° C.) solution of (4-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl) phenyl) methanol (950 mg, 2.42 mmol, 1 equiv) in anhydrous dichloromethane (100 mL) was added Dess-Martin periodinane (1.23 g, 2.90 mmol, 1.2 equiv) and the mixture was stirred for 2 h at room temperature. The reaction mixture was cooled to 0° C. and quenched with saturated aqueous sodium bicarbonate. The resulting mixture was extracted with dichloromethane (100 mL×2). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude residue was purified by silica gel column chromatography using a 0-5% gradient of methanol in dichloromethane to afford 4-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl) benzaldehyde (650 mg, 65%) as a brown solid. MS (ESI) calculated for C24H17N5O: 391.14 m/z, found 392.25 [M+H]+.
A mixture of 4-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl) benzaldehyde (200 mg, 0.48 mmol, 1 equiv), 4-(4-((tert-butyldimethylsilyl) oxy)-3-(1,3-dioxolan-2-yl) phenyl) piperidine (Intermediate 136-1) (188 mg, 0.524 mmol, 1.1 equiv) and sodium cyanoborohydride (299 mg, 4.76 mmol, 10 equiv) in methanol (10 mL) was stirred at room temperature for 1 hour. The reaction was quenched with water (20 mL) and extracted with ethyl acetate (20 mL×3). The combined organic extracts were washed with brine, filtered, and concentrated in vacuo. The resulting residue was purified by flash column chromatography on silica gel using a 0-100% gradient of ethyl acetate in petroleum ether to afford 3-(3-(4-((4-(4-((tert-butyl dimethylsilyl) oxy)-3-(1,3-dioxolan-2-yl) phenyl) piperidin-1-yl) methyl) phenyl)-5-phenyl-3H-imidazo[4,5-b]pyridin-2-yl) pyridin-2-amine (171 mg, 49%) as a white solid. MS (ESI) calculated for C44H50N6O3Si: 738.37 m/z, found 739.40 [M+H]+.
Triethylamine trihydrofluoride (1 mL) was added to a solution of 3-(3-(4-((4-(4-((tert-butyl dimethylsilyl) oxy)-3-(1,3-dioxolan-2-yl) phenyl) piperidin-1-yl) methyl) phenyl)-5-phenyl-3H-imidazo[4,5-b]pyridin-2-yl) pyridin-2-amine (100 mg, 0.13 mmol, 1 equiv) in tetrahydrofuran (10 mL). The mixture was stirred at room temperature for 1 hour. The mixture was concentrated in vacuo and the resulting residue was purified by preparative HPLC on a XBridge Prep OBD C18 Column using a 17-42% gradient of acetonitrile in water (+10 mmol/L ammonium bicarbonate) to afford 5-(1-(4-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl) benzyl) piperidin-4-yl)-2-hydroxybenzaldehyde (Example 136) (10.3 mg, 10.96%) as an off-white solid. MS (ESI) calculated for C36H31N6O2: 580.26 m/z, found 581.25 [M+H]+. 1H NMR (300 MHz, methanol-d4) δ (ppm) 8.30 (d, J=8.5 Hz, 1H), 8.10-7.98 (m, 4H), 7.85-7.69 (m, 4H), 7.69-7.62 (m, 1H), 7.55-7.36 (m, 4H), 7.28 (s, 1H), 7.11 (d, J=8.3 Hz, 1H), 6.82 (dd, J=13.6, 8.5 Hz, 1H), 6.71 (dd, J=7.7, 5.7 Hz, 1H), 5.62 (d, J=6.5 Hz, 1H), 4.49 (s, 2H), 3.69 (d, J=12.0 Hz, 2H), 3.23 (d, J=12.5 Hz, 2H), 2.89 (d, J=15.5 Hz, 1H), 2.14 (d, J=14.4 Hz, 2H), 1.98 (t, J=12.9 Hz, 2H). Note that the aldehyde signal is not visible for some compounds dissolved in methanol-d4.
A mixture of 4-bromo-2-(1,3-dioxolan-2-yl)phenoxy(tert-butyl)dimethylsilane (Intermediate 131-1) (500 mg, 1.39 mmol, 1 equiv), benzyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylate (525 mg, 1.53 mmol, 1.1 equiv), potassium carbonate (577 mg, 4.17 mmol, 3 equiv) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (102 mg, 0.139 mmol, 0.1 equiv) in 1,4-dioxane (15 mL) and water (0.1 mL) was stirred at 80° C. for 16 h under nitrogen atmosphere. The reaction mixture was cooled to room temperature and quenched with water (20 mL). The resulting mixture was extracted with ethyl acetate (20 mL×3). The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using a 0-100% gradient of ethyl acetate in petroleum ether to afford benzyl 4-(4-((tert-butyldimethylsilyl)oxy)-3-(1,3-dioxolan-2-yl)phenyl)piperidine-1-carboxylate (300 mg, 43%) as a yellow solid. MS (ESI) calculated for C28H39NO5Si: 497.26 m/z, found 498.20 [M+H]+.
A solution of benzyl 4-(4-((tert-butyldimethylsilyl)oxy)-3-(1,3-dioxolan-2-yl)phenyl)piperidine-1-carboxylate (200 mg, 0.403 mmol, 1 equiv) in methanol (10 mL) was treated with 10% palladium on carbon (15 mg, 0.14 mmol, 0.35 equiv) and the resulting mixture was stirred at room temperature under hydrogen atmosphere for 2 h. The resulting mixture was filtered rinsing with methanol (20 mL×5). The filtrate was concentrated under reduced pressure to afford 4-(4-((tert-butyldimethylsilyl)oxy)-3-(1,3-dioxolan-2-yl)phenyl)piperidine (Intermediate 136-1) (120 mg, 812%) as yellow oil, which was used in subsequent transformations without further purification. MS (ESI) calculated for C20H33NO3Si: 363.22 m/z, found 364.20 [M+H]+.
4-[2-(2-Aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]benzoic acid (Intermediate 29-2) (100 mg, 0.245 mmol, 1 equiv) and N,N-dimethylaminopyridine (0.28 g, 2.3 mmol, 5 equiv) were added to a solution of 1-{3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)phenyl}methanamine (Intermediate 137-1) (91 mg, 0.29 mmol, 1.2 equiv), N,N-diisopropylethylamine (159 mg, 1.23 mmol, 5 equiv) and HATU (187 mg, 0.490 mmol, 2 equiv) in N,N-dimethylformamide (2 mL). The resulting mixture was stirred at room temperature for 1 h. The reaction was quenched with water (10 mL) and extracted with ethyl acetate (10 mL×3). The combined extracts were washed with water and brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by flash column chromatography on silica gel using a 0-100% gradient of ethyl acetate in petroleum ether to afford 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]-N-({3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)phenyl}methyl)benzamide (80 mg, 47%) as a white solid. MS (ESI) calculated for C40H42N6O4Si: 698.30 m/z, found 699.20 [M+H]+.
4-(2-(2-aminopyridin-3-yl)-5-phenyl-3H-imidazo[4,5-b]pyridin-3-yl)-N-(3-((tert-butyldimethylsilyl)oxy)-4-(1,3-dioxolan-2-yl)benzyl)benzamide (80 mg, 0.11 mmol, 1 equiv) was dissolved in dichloromethane (5 mL) and 2,2,2-trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated in vacuo and purified by preparative HPLC on a YMC-Actus Triart C18 ExRS column using a 17-42% gradient of acetonitrile in water (+10 mmol/L ammonium bicarbonate) to afford 4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]-N-[(4-formyl-3-hydroxyphenyl)methyl]benzamide (Example 137) (8.7 mg, 14%) as a yellow solid. MS (ESI) calculated for C32H24N6O3: 540.19 m/z, found 541.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm): 10.02 (s, 1H), 8.23 (d, J=8.4 Hz, 1H), 8.12-7.95 (m, 6H), 7.75-7.61 (m, 3H), 7.51-7.36 (m, 4H), 7.07 (d, J=8.3 Hz, 1H), 6.99 (s, 1H), 6.56 (in, J=7.7, 5.0 Hz, 1H), 4.64 (s, 2H).
To a solution of 4-formyl-3-hydroxybenzonitrile (7.31 g, 49.7 mmol, 1 equiv) in toluene (100 mL) was added p-toluenesulfonic acid (0.86 g, 5.0 mmol, 0.1 equiv), ethylene glycol (15.44 g, 248.7 mmol, 5 equiv) and triethyl orthoformate (22.12 g, 149.2 mmol, 3 equiv). The resulting solution was stirred at room temperature for 10 min then at 90° C. for 18 h. The resulting mixture was cooled to 0° C. and quenched by the addition of saturated aqueous ammonium chloride (80 mL) The resulting mixture was extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with water (100 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-7% gradient of ethyl acetate in petroleum ether to afford 4-(1,3-dioxolan-2-yl)-3-hydroxybenzonitrile (5.0 g, 53%) as a light-yellow oil. MS (ESI) calculated for C10H9NO3: 191.06 m/z, found 192.10 [M+H]+.
To a solution of 4-(1,3-dioxolan-2-yl)-3-hydroxybenzonitrile (1.48 g, 7.75 mmol, 1 equiv) and imidazole (1.05 g, 15.5 mmol, 2 equiv) in dichloromethane (200 mL) was added tert-butyldimethylsilyl chloride (1.64 g, 10.9 mmol, 1.4 equiv). After stirring overnight at room temperature, the reaction was quenched by the addition of water (200 mL) at 0° C. The resulting mixture was extracted with ethyl acetate (300 mL×3). The combined organic layers were washed with water (300 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using a 0-7% gradient of ethyl acetate in petroleum ether to afford 3-((tert-butyldimethylsilyl)oxy)-4-(1,3-dioxolan-2-yl)benzonitrile (1.8 g, 79%) as a colorless oil. MS (ESI) calculated for C16H23NO3Si: 305.14 m/z, found 306.15 [M+H]+.
A solution of 3-[(tert-butyldimethylsilyl)oxy]-4-(1,3-dioxolan-2-yl)benzonitrile (200 mg, 0.655 mmol, 1 equiv) in methanol (10 mL) was treated with 10% palladium on carbon (15 mg, 0.14 mmol, 0.46 equiv) and the resulting mixture was stirred at room temperature under hydrogen atmosphere overnight. The resulting mixture was filtered, rinsing with methanol (20 mL×5). The filtrate was concentrated under reduced pressure to afford (3-((tert-butyldimethylsilyl)oxy)-4-(1,3-dioxolan-2-yl)phenyl)methanamine (Intermediate 137-1) (160 mg, 79%) as yellow oil, which was used in subsequent transformations without further purification. MS (ESI) calculated for C1H27NO3Si: 309.18 m/z, found 310.20 [M+H]+.
A mixture of 3-{3-[4-(Aminomethyl)phenyl]-5-phenylimidazo[4,5-b]pyridin-2-yl}pyridin-2-amine (Intermediate 4-1) (170 mg, 0.433 mmol, 1 equiv), 4-formyl-3-hydroxybenzoic acid (108 mg, 0.650 mmol, 1.5 equiv), HATU (247 mg, 0.650 mmol, 1.5 equiv) and N,N-diisopropylethylamine (0.4 mL, 2.3 mmol, 5.3 equiv) in N,N-dimethylformamide (5 mL) was stirred at room temperature for 1 h. The reaction was quenched with water (5 mL). The resulting mixture was extracted with ethyl acetate (3×50 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue obtained was purified by preparative HPLC on a XBridge Shield RP18 OBD Column using a 33-58% gradient of acetonitrile in water (+10 mmol/L ammonium bicarbonate) to afford N-({4-[2-(2-aminopyridin-3-yl)-5-phenylimidazo[4,5-b]pyridin-3-yl]phenyl}methyl)-4-formyl-3-hydroxybenzamide (Example 138) (5.1 mg, 2.2%) as a yellow solid. MS (ESI) calculated for C32H24N6O3: 540.19 m/z, found 541.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ (ppm) 10.31 (s, 1H), 9.30-9.23 (m, 1H), 8.28-8.22 (m, 1H), 8.06-7.92 (m, 4H), 7.74-7.70 (m, 1H), 7.51-7.49 (m, 1H), 7.49-7.37 (m, 8H), 7.42-7.27 (m, 1H), 7.23-7.17 (m, 1H), 6.93 (s, 2H), 6.43-6.38 (m, 1H), 4.61-4.56 (m, 2H).
Example 139 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 139-1 in place of the amine starting material and Intermediate 21-2 in place of Intermediate 1-4. MS (ESI) calculated for C30H25FN6O3: 536.20 m/z, found 537.20 [M+H]+. 1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.19 (s, 1H), 8.99 (s, 1H), 8.11-8.13 (m, 1H), 8.01-8.03 (m, 1H), 7.67-7.70 (m, 1H), 7.43-7.46 (m, 1H), 7.30-7.36 (m, 3H), 7.22-7.24 (m, 2H), 6.91-6.95 (m, 1H), 6.73-6.77 (m, 1H), 3.64 (s, 2H), 2.50-2.52 (m, 3H), 1.21-1.30 (m, 4H). 19F-NMR (400 MHz, DMSO-d6) δ (ppm): −139.30.
Intermediate 139-1 was prepared in a manner analogous to Intermediate 10-1 (starting from Step 2) using 6-methyl-3-nitropyridin-2-amine in place of the amine starting material and tert-butyl N-[1-(4-bromophenyl)cyclopropyl]carbamate in place of tert-butyl N-[(4-bromophenyl)methyl]carbamate. MS (ESI) calculated for C21H20N6: 356.17 m/z, found 357.15 [M+H]+.
Example 140 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 140-2 in place of the amine starting material and Intermediate 21-1 in place of Intermediate 1-4. MS (ESI) calculated for C33H30FN7O4: 607.23 m/z, found 608.20 [M+H]+. 1H-NMR (400 MHz, DMSO-d6) δ (ppm): 10.19 (s, 1H), 7.91-8.09 (m, 2H), 7.39-7.59 (m, 2H), 7.29-7.38 (m, 2H), 7.15-7.21 (m, 2H), 6.85-6.99 (m, 2H), 6.70-6.79 (m, 1H), 3.65-3.76 (m, 6H), 3.30-3.42 (m, 4H), 1.15-1.35 (m, 4H). 19F-NMR (400 MHz, DMSO-d6) δ (ppm): −139.29.
Intermediate 140-2 was prepared in a manner analogous to Intermediate 10-1 (starting from Step 2) using Intermediate 140-1 in place of the amine starting material and tert-butyl N-[1-(4-bromophenyl)cyclopropyl]carbamate in place of tert-butyl N-[(4-bromophenyl)methyl]carbamate. MS (ESI) calculated for C24H25N7O: 427.21 m/z, found 428.20 [M+H]+.
To a solution of 6-bromo-3-nitropyridin-2-amine (2.0 g, 9.2 mmol, 1 equiv) in dimethyl sulfoxide (16 mL) was added morpholine (7.99 g, 91.6 mmol, 10 equiv). The resulting mixture was stirred at 140° C. for 1 h. The mixture was concentrated in vacuo to afford 6-(morpholin-4-yl)-3-nitropyridin-2-amine (Intermediate 140-1) (1.92 g, 93%) as a yellow solid, which was used without purification in subsequent transformations. MS (ESI) calculated for C9H12N4O3: 224.09 m/z, found 225.05 [M+H]+.
Example 141 was prepared in a manner analogous to Example 1 (starting from Step 3) using Intermediate 141-2 in place of the amine starting material and Intermediate 21-1 in place of Intermediate 1-4. MS (ESI) calculated for C32H25FN803: 588.20 m/z, found 589.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ (ppm): 10.12 (s, 1H), 8.38-8.46 (m, 1H), 8.29-8.32 (m, 1H), 7.91-8.05 (m, 2H), 7.75-7.87 (m, 2H), 7.39-7.51 (m, 3H), 7.19-7.23 (m, 2H), 6.88-6.92 (m, 1H), 6.79-6.82 (m, 1H), 6.47-6.53 (m, 1H), 3.55-3.63 (m, 2H), 1.14-1.33 (m, 4H).
Intermediate 141-2 was prepared in a manner analogous to Intermediate 10-1 (starting from Step 2) using Intermediate 141-1 in place of the amine starting material and tert-butyl N-[1-(4-bromophenyl)cyclopropyl]carbamate in place of tert-butyl N-[(4-bromophenyl)methyl]carbamate. MS (ESI) calculated for C23H20N8: 408.18 m/z, found 409.20 [M+H]+.
A suspension of 6-bromo-3-nitropyridin-2-amine (2.0 g, 9.2 mmol), pyrazole (937 mg, 13.8 mmol), EPhos (736 mg, 1.38 mmol), EPhos Pd G4 (843 mg, 0.918 mmol) and cesium carbonate (6.0 g, 18 mmol) in 1,4-dioxane (20 mL) was stirred at 100° C. for 2 h under nitrogen atmosphere. The resulting mixture was cooled to room temperature and diluted with water (100 mL). The resulting mixture was extracted with ethyl acetate (100 mL×3). The combined extracts were concentrated under vacuum and the resulting oil was purified by silica gel column chromatography eluting with dichloromethane to afford 3-nitro-6-(pyrazol-1-yl)pyridin-2-amine (Intermediate 141-1) as brown solid (1.72 g, 91%). MS (ESI) calculated for C8H7N5O2: 205.06 m/z, found 206.05 [M+H]+.
Unless specified otherwise, the following protein sequences and protein expression protocols were used for experimental data collection. Amino acid mutation numbers are based on the naturally occurring human AKT1 amino acid sequence and not the genetically modified sequence (e.g., the E17K mutation is shown as the 29th residue in the following sequences).
The human AKT1 (WT or E17K) with the N-terminal affinity 6-His-tag followed by a cleavable TEV-tag was expressed in Sf9 insect cells using baculovirus-insect cell expression system, which were infected in 2 million per milliliter at 300 K. The cells were harvested after an incubation time of 72 hours at 300 K and stored at 193 K until lysis.
The cell paste from 1 L Sf9 cell pellet were lysed in (50 mM Tris, 500 mM NaCl, 1 mM TCEP, 10% glycerol, 0.1% Triton X-100, pH 8.0) supplemented with cOmplete EDTA-free protease inhibitor tablet (Roche). Purification was performed by Ni-NTA capture of the His-tagged protein in buffer A, washed with (50 mM Tris, 500 mM NaCl, 1 mM TCEP, 10% glycerol, pH 8.0, 20 mM imidazole), and eluted in two steps with (50 mM Tris, 500 mM NaCl, 1 mM TCEP, 10% glycerol, pH 8.0, 50 and 250 mM imidazole). Fraction eluted with 50 mM imidazole was dialyzed against 4 L dialysis buffer and digested by His-tagged TEV (TEV:Protein=1:15) at 4° C. overnight and then loaded onto 2nd Ni-NTA column. The flow through were pooled and diluted to 50 mM NaCl, and then load onto Superdex 75 100/300 GL column, separately using SEC buffer (25 mM Tris, 100 mM NaCl, 5 mM DTT, 10% glycerol, pH 7.5). The tag cleaved AKT1 (2-480, WT or E17K) were concentrated in final storage buffer (25 mM Tris, 100 mM NaCl, 5 mM DTT, 10% glycerol, pH 7.5) and stored in −80° C.
AKT1 WT or AKT1 E17K (1 μM) was treated with a compound from Table 1 (1.5 μM) for 2 hrs at 37° C., with and without ARQ092 (50 μM) competitor as a specificity control. Reversible imines were captured by reductive amination with 5 mM NaBH4 for 5 min. Reduction reactions were quenched by diluting the sample in equal volume acetonitrile+1% TFA, then analyzed by LCMS.
AKT1 WT or AKT1 E17K (1 μM) were treated with a compound of Table 1 (1.5 μM) for 2 hours at 37° C., then ARQ092 (50 μM final concentration) was added to prevent aldehyde-containing compound rebinding after initial dissociation. At various timepoints, aliquots were removed and quenched with 5 mM NaBH4 for 5 min to capture the reversible imine through reductive amination. Reduction reactions were quenched by diluting the sample in equal volume acetonitrile+1% TFA, then analyzed by LCMS. Deconvoluted monoisotopic intensities of unmodified vs. compound-modified mass peaks were compared.
The following protein sequences were used for the described crystallography studies. Amino acid mutation numbers are based on the naturally occurring human AKT1 amino acid sequence and not the genetically modified sequence (e.g., the E17K mutation is shown as the 34th residue in the following sequences).
AKT1 WT and AKT1 E17K were gene synthesized and cloned by Gibson assembly into the pFastBac vector. The plasmid was transformed into DH10Bac cells and the recombinant bacmid was isolated and used to generate baculovirus in Sf9 cells.
Expression of AKT1 was induced in Sf9 cells by infection of 1.0 L of cultured cells (1.1 M/mL) with 20 mL of baculovirus solution and incubated for 72 h. The cells were harvested (3500 rpm, 10 min), and the pellets were resuspended in 100 mL lysis buffer (25 mM HEPES, pH 8.0, 150 mM NaCl, 1.0 mM TCEP) supplemented with protease inhibitor cocktail (Roche). The cells were lysed by sonication and the cell debris pelleted by centrifugation (20000 rpm, 30 min). The clarified lysate was incubated with NiNTA (3.0 mL) and the resin was washed with lysis buffer (3×10 mL) and then 5 mL wash buffer (40 mM imidazole, 25 mM HEPES, pH 8.0, 150 mM NaCl, 1.0 mM TCEP). AKT1 was eluted in 6 mL of elution buffer (400 mM imidazole, 25 mM Hepes, pH 8.0, 150 mM NaCl, 1.0 mM TCEP). To the eluted protein was added 1.5 mg TEV protease. After 1 hour, LCMS showed complete TEV cleavage. The resulting solution was diluted to 60 mL with 20 mM Tris pH 8.0, 150 mM NaCl and then passed through Ni-NTA (2 mL) and the flowthrough collected. The resin was further washed with 5 mL of 25 mM Tris pH 7.5, 100 mM NaCl, 40 mM imidazole buffer to recover the remaining AKT1 which was combined with the flowthrough. The protein was buffer exchanged into 20 mM Tris pH 8.0 20 mM NaCl and purified by ion exchange chromatography (Q HiTrap, gradient from 20 to 250 mM NaCl). The resulting protein was purified further by gel filtration on a Superdex 200 (16/60) column (gel filtration buffer: 25 mM HEPES pH 8.0, 150 mM NaCl, 1 mM TCEP). The protein was then concentrated to 5 mg/mL, flash-frozen in liquid nitrogen and stored at −80° C. The final yield was ˜4 mg AKT1.
NB41 was gene synthesized and cloned by Gibson assembly into a PET26b vector containing an N-terminal PelB signal sequence. BL21 cells were transformed with the NB41 PET26b plasmid. The transformed bacteria were grown in 2 L TB+kanamycin at 37° C. until it reached OD600=0.66. The culture was cooled to 18° C., induced with 0.25 mM IPTG and expressed overnight at 18° C. The bacteria were harvested (5000 rpm, 10 min) and the resulting pellet frozen.
The pellet was warmed to 4° C. and lysed in 15 mL TS buffer (TS buffer=200 mM Tris pH 8.0, 500 mM sucrose) with rotation. After 1 hour, 30 mL of TS/4 buffer was added (TS/4 buffer=50 mM Tris, 125 mM Sucrose). After a further 45 min rotation at 4° C., the sample was centrifuged for 30 min at 7500 g. The supernatant was collected and passed over 3 mL NiNTA resin. The resin was washed with 3×10 mL wash buffer (40 mM imidazole, 25 mM HEPES, pH 8.0, 150 mM NaCl). NB41 was eluted in 6 mL of elution buffer (400 mM imidazole, 25 mM Hepes, pH 8.0, 150 mM NaCl). The resulting protein was purified further by gel filtration on a Superdex 75 (16/60) column (gel filtration buffer: 20 mM Tris pH 7.5, 100 mM NaCl, 1 mM TCEP). The protein was then concentrated to 5 mg/mL and flash-frozen in liquid nitrogen and stored at −80° C.
To 1 mg of AKT1 WT or AKT1 E17K in 10 mL of 20 mM HEPES (pH 8.0), 150 mM NaCl, 1 mM TCEP buffer was added 200 μL Compound 110 or Compound 133 (0.5 mM stock in DMSO) to give a final concentration of 10 μM. After 10 min, the sample was concentrated to 1 mL and then 0.55 mg NB41 was added and incubated for 15 min. The resulting complex was purified by gel filtration on a Superdex 200 (16/60) column (gel filtration buffer: 20 mM HEPES pH 8.0, 20 mM NaCl, 1 mM TCEP). The protein was then concentrated to 6.2-7.5 mg/mL and used directly for crystal formation.
AKT1-E17K/Compound 133: AKT1 E17K at 3 mg/ml were incubated with Compound 133 at 1:5 molar ratio on ice for 1 hr. Co-crystals of AKT1-E17K/Compound 133 were obtained at 277K from hanging drops using reservoir solutions containing 1.25 mM Na-acetate 6.0, 3.75 mM sodium citrate 7.5, 15% PEG MME 2000. Co-crystals were flash cooled in liquid nitrogen after addition of 20% ethylene glycol supplemented to the mother liquor. Diffraction data of AKT1-E17K/Compound 133 was obtained at 100 K at beamline 08B1 in CLS, using an Pilatus3 S 6M detector (DECTRIS Ltd.). These datasets were further processed with autoPROC (Global Phasing). The initial structures were determined through molecular replacement with Phaser in CCP4 suite with the structure of PDB ID 5kcv as a search model. A chemical restraint dictionary was generated using ACEDRG and CCP4i2. The models were manually adjusted using Coot, refined with REFMAC5 to a final resolution of 3.0 Å for the AKT1-E17K/Compound 133 complex.
AKT1-WT/Compound 133-NB41: The protein solution: AKT1-WT/Compound 133-NB41 (7.5 mg/mL) in 20 mM HEPES (pH 8.0), 20 mM NaCl, 1 mM TCEP, 0.4 mM iodoacetamide; and the reservoir solution: 19% PEG3350, 200 mM Li2SO4, 100 mM BisTris Propane pH 7.2, 10% ethylene glycol; were mixed in a 1:1 ratio and the crystals grown overnight and were cryoprotected in the reservoir solution containing 25% ethylene glycol.
AKT1-E17K/Compound 110-NB41: The protein solution: AKT1-E17K/Compound 110-NB41 (6.2 mg/mL) in 20 mM HEPES (pH 8.0), 20 mM NaCl, 1 mM TCEP; and reservoir solution: 18% PEG3350, 200 mM Li2SO4, 100 mM BisTris Propane pH 7.8, 10% ethylene glycol; were mixed in a 1:1 ratio and the crystals grown overnight were cryoprotected in the reservoir solution containing 25% ethylene glycol.
Diffraction data were collected at beamline 2.0.1 of the Advanced Light Source (Lawrence Berkeley National Laboratory) and indexed and integrated using iMosflm, scaled with Scala and solved by molecular replacement using Phaser. The structure was manually refined with Coot and PHENIX.
Statistics for the highest-resolution shell are shown in parentheses.
Live cell target engagement assays were performed as described (Vasta, J. D. et al Cell Chem Biol 2018, 25(11), 206-214). Briefly, HEK293 cells were transfected with plasmids encoding kinase-NanoLuciferase fusion proteins overnight (Promega). Cells were then treated with serial dilutions of compound and an ˜EC50 concentration of fluorescently-tagged ATP-competitive tracer (Promega). After incubation for 2 hours at 37° C., luciferase substrate and extracellular luciferase inhibitor were added to all wells, and luminescent intensity at 460 nm and 600 nm were measured on a multimode plate-reader. The ratio of E600/E460 was calculated to give the tracer engagement signal (BRET). IC50 values were calculated by fitting BRET values to a log(inhibitor) vs. response Hill equation.
Covalent labelling selectivity were measured by intact-protein mass spectrometry. AKT1(WT) or AKT1(E17K) (1 μM) was incubated with inhibitor (5 μM) for 15 minutes. The covalent imine adducts were reduced with 5 mM NaBH4 and the percentage labelling determined by LCMS.
Identification of the sites of covalent modification by tandem mass spectrometry: AKT1(E17K) (5 μM) was incubated with excess compound (Compound A) (50 μM) for 15 minutes and then treated with NaBH4 (5 mM) to capture the reversible imine through reductive amination. The labelled protein was digested with trypsin and the peptides analyzed by tandem mass spectrometry. MS/MS spectra were searched using a database comprised of the peptide sequence for the recombinant AKT1, along with the reverse sequence as decoy. In addition to typical variable modifications, a variable modification on lysine corresponding to compound addition (minus H2O) was allowed (e.g. 564.605 Da: 18 Da=546.605 Da). Peptides bearing lysine residues modified with Compound A were identified by spectral matching. Parent Ion: [GK(Compound A)YIKTWRPR]+5 m/z=371.3965 (z=5); −0.69 ppm.
Table 5 provides the calculated ion fragments for the covalent modification of K17 in AKT1(E17K) by Compound A.
Table 6 provides the calculated ion fragments for the covalent modification of K297 in AKT1(E17K) by Compound A.
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Determination of compound dissociation rate by intact protein mass spectrometry:Recombinant AKT1 containing appropriate mutations (1 μM) was treated with inhibitor (5 μM) at room temperature. After 15 minutes, ARQ-092 (50 μM final concentration) was added to prevent aldehyde-containing compound rebinding after initial dissociation. At various timepoints, aliquots were removed and quenched with 5 mM NaBH4 to capture the reversible imine through reductive amination, then analyzed by LCMS. Deconvoluted monoisotopic intensities of unmodified vs. compound-modified mass peaks were compared.
Identification of site of covalent modification by tandem mass spectrometry: AKT1(WT) or AKT1(E17K) (5 μM) was incubated with excess compound (Compound A or Compound B) (50 μM) for 15 minutes and then treated with NaBH4 (5 mM) to capture the reversible imine through reductive amination. The labelled protein was digested with trypsin and the peptides analyzed by tandem mass spectrometry. MS/MS spectra were searched using a database comprised of the peptide sequence for the recombinant AKT1, along with the reverse sequence as decoy. In addition to typical variable modifications, a variable modification on lysine corresponding to compound addition (minus H2O) was allowed (e.g. 564.605 Da: 18 Da=546.605 Da).
Measurement of target engagement durability in intact cells: Beas-2b cells stably over-expressing either Flag-AKT1 WT or Flag-AKT1 E17K were treated with compound for 30 minutes at 37 C. The media was then aspirated and replaced with fresh media and the cells were incubated at 37° C. At various timepoints, cells were washed with PBS and then lysed in buffer containing 5 mM NaBH4. After 1 h, the lysate was precipitated in MeOH/CHCl3, resuspended in 1% SDS in PBS and then subjected to copper catalyzed click chemistry with TAMRA-biotin-azide. The samples were precipitated again and then resuspended in 1.5× SDS loading buffer, separated by SDS-PAGE and analyzed by in-gel fluorescence and western blot.
Measurement of phospho-AKT inhibition and treatment durability in intact cells: Cells were treated for 2 hrs with compound, then washed twice with fresh media with a 5 min 37 C incubation between washes. After additional incubation for 0, 8 or 24 h at 37 C, cells were washed with PBS and lysed in buffer containing protease and phosphatase inhibitors. Lysates were analyzed by SDS-PAGE/western blot using standard methods.
Determining live cell AKT1 E17K, AKT1 WT, and AKT2 target engagement by NanoBRET competitive probe displacement: Live cell target engagement assays were performed as described (Vasta, J. D. et al. Cell Chem Biol 2018, 25(11), 206-214). Briefly, HEK293 cells were transfected with plasmids encoding kinase-NanoLuciferase fusion proteins overnight. Cells were then treated with serial dilutions of compound and an ˜EC50 concentration of fluorescently-tagged ATP-competitive tracer (Promega). After incubation for 2 h at 37 C, luciferase substrate and extracellular luciferase inhibitor were added to all wells, and luminescent intensity at 460 nm and 600 nm were measured on a multimode plate-reader. The ratio of E600/E460 was calculated to give the tracer engagement signal (BRET).
Table 7 provides IC50 values for Compound A, Compound B, and Miransertib (ARQ-092). Table 7 includes IC50 values for the selected compounds; with compounds having a IC50 of less than or equal to 100 nM as A; 100 nM>B≥500 nM as B; and greater than 500 nM as C.
AKT1(E17K or WT) (1 μM) was incubated with Compound A or Compound B (5 μM) for 15 minutes in 25 mM HEPES pH 8.0, 150 mM NaCl buffer. Then ARQ-092 (50 μM) was added and aliquots quenched with 5 mM NaBH4 at a series of timepoints and the percentage labelling determined by LCMS.
This application is the by-pass continuation of International Application No. PCT/US2023/063513, filed Mar. 1, 2023, which claims the benefit of U.S. Provisional Application No. 63/315,933, filed Mar. 2, 2022, the entire contents of each of which are incorporated herein by reference.
| Number | Date | Country | |
|---|---|---|---|
| 63315933 | Mar 2022 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/US2023/063513 | Mar 2023 | WO |
| Child | 18819786 | US |
