Patent Application
20240400545
The present invention relates to the field of medicinal chemistry; specifically, the present invention relates to a novel class of derivatives containing a tricyclic heteroaryl group, its synthesis and its application as a KAT6 inhibitor in the preparation of drugs for use in the treatment of a wide range of diseases related to tumors and the like.
Histone acetyltransferases use acetyl-CoA to transfer acetyl groups to lysine residues of histone or other substrate proteins. Acetylation is a reversible post-translational modification that plays an important role in eukaryotic gene expression, cell cycle, and signaling pathway transduction. According to homology, histone acetyltransferases are divided into four families, namely GNAT (Gcn5-related acetyltransferase) family, p300/CBP family, SRC/p160 family, and MYST (MOZ, Ybf2/Sas3, Sas2 and Tip60) family. The acetyltransferase activity of MYST family is affected by the MYST domain (catalytic domain). The MYST domain contains an acetyl-CoA-binding motif and a rare C2HC-type zinc finger structure, where the acetyl-CoA-binding motif is structurally conserved with other histone acetyltransferases. The MYST domain is the defining feature of MYST family proteins.
KAT6A (Lysine Acetyltransferase 6A, also known as MOZ) and KAT6B (Lysine Acetyltransferase 6B, also known as MORF) are acetyltransferases belonging to the MYST family. KAT6A is chromosomally translocated in acute myeloid leukemia and has amplification mutations in lung cancer, breast cancer, ovarian cancer, endometrial cancer, bladder cancer, and esophageal cancer. Similarly, KAT6B is also chromosomally translocated in a variety of cancer types. MOZ- and MORF-linked fusion proteins identified in acute myeloid leukemia include MOZ-CBP, MOZ-p300, MOZ-TIF2, MOZ-NcoA3, MOZ-LEUTX, and MORF-CBP. MOZ-TIF2 has transforming activity in cultured cells and can induce acute myeloid leukemia in mice. In tumor cells with KAT6A and KAT6B amplification, the expression of KAT6A and KAT6B is closely related to the gene copy number, indicating that there is selective pressure to maintain their activity during tumorigenesis. In addition, in cell proliferation tests, tumor cells with high expression of KAT6A and KAT6B are usually more dependent on the activity of KAT6A and KAT6B.
KAT6A and KAT6B generally extensively acetylate the lysine 23 site of histone H3 (H3K23), and KAT6A can also modify the acetylation of the H3K9 site of its regulatory gene. KAT6A interacts with some transcription factors such as p53 and RUNX1 to acetylate histones and regulate the expression of downstream genes. KAT6A binds to the proximal promoter region of the estrogen receptor ERα (Estrogen Receptor a) to activate the expression of ERα genes. In ER+, KAT6A amplification or high expression breast cancer cells, inhibition of KAT6A acetyltransferase activity or knockdown of KAT6A can significantly inhibit the proliferation of breast cancer cells. In addition, KAT6A acetyltransferase activity is essential for promoting the expression of MEIS1 and HOXa9 genes, which are often overexpressed in some lymphoma and leukemia cells. In MYC-induced lymphoma mouse model, the deletion of a KAT6A allele can significantly prolong the median survival of mice. In mice, KAT6B allele mutations lead to a significant reduction in the division and differentiation of cortical progenitors, which seriously affects the development of the cerebral cortex. KAT6B also plays an important role in maintaining the number of adult neural stem cells. KAT6B is also mutated in some rare types of leukemia.
Based on the above background, we developed a novel series of inhibitors of KAT6.
It is an object of the present invention to provide a novel class of KAT6 inhibitors.
In the first aspect of the present invention, a compound of the following formula (I), or the optical isomers, pharmaceutically acceptable salts, prodrugs, deuterated derivatives, hydrates, or solvates thereof is provided:
In another preferred embodiment, the compound (I) is of formula (II):
In another preferred embodiment, said E is selected from C3-6 cycloalkyl, 3- to 6-membered heterocyclyl, C3-6 cycloalkyl-C1-4 alkyl, and 3- to 6-membered heterocyclyl-C1-4 alkyl; each R3 is independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 haloalkyl, CN, ORb, —C(O)Rg; or two R3 attaching to the same carbon atom of the cycloalkyl or heterocyclyl ring together with the carbon atom form C=T; wherein, T is selected from O and CR12R13; wherein, R12 and R13 are each independently selected from hydrogen, fluorine, and C1-4 alkyl; the alkyl on said R12 or R13 is optionally substituted by one or more groups selected from the group consisting of halogen, C1-4 haloalkyl, ORb, and NRbRb; each Rb is independently selected from hydrogen, C1-4 alkyl, or C1-4 haloalkyl; R9 is selected from hydrogen, C1-4 alkyl, and C3-8 cycloalkyl.
In another preferred embodiment, the compound (I) is of formula (III):
In another preferred embodiment, the compound (I) is of formula (IV):
In another preferred embodiment, the compound (I) is of formula (V):
In another preferred embodiment, the compound (I) is of formula (VI):
In another preferred embodiment, the compound (I) is of formula (VII):
In another preferred embodiment, the compound (I) is of formula (VIII):
In another preferred embodiment, the R4 is selected from the group consisting of hydrogen, C1-4 alkyl, C3-8 cycloalkyl, 3- to 9-membered heterocyclyl, aryl, and heteroaryl; wherein said alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted by one or more groups selected from the group consisting od halogen, CN, ORa, —C(O)Rm, —C(O)ORa, —S(O)2Rm, C1-4 alkyl, C3-6 cycloalkyl, 3- to 8-membered heterocyclyl, aryl, and heteroaryl; or said cycloalkyl or heterocyclyl is substituted with =T; wherein each Ra is as defined above; Rm is selected from the group consisting of C1-4 alkyl, C2-4 alkenyl, C2-4 alkynyl, and C3-6 cycloalkyl; and T is as defined above.
In another preferred embodiment, the R2 is each independently selected from the group consisting of hydrogen, halogen, C1-4 alkyl, C1-4 haloalkyl, ORa, C3-6 cycloalkyl, and 3- to 8-membered heterocyclyl.
In another preferred embodiment, the R7 is each independently selected from hydrogen, halogen, C1-4 alkyl, C1-4 haloalkyl, ORa, SRa, NRaRa, C3-6 cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, or heteroaryl C1-4 alkyl; each Ra is independently selected from hydrogen and C1-4 alkyl.
In another preferred embodiment, the compound (I) is of formula (IX):
In another preferred embodiment, the compound (I) is of formula (X):
In another preferred embodiment, the compound (I) is of formula (IX):
In another preferred embodiment, the compound (I) is of formula (XII):
In another preferred embodiment, the compound (I) is of formula (VII):
In the second aspect of the present invention, a compound of the structure shown in formula (XIV), or an optical isomer, a pharmaceutically acceptable salt, a prodrug, a deuterated derivative, a hydrate, or a solvent thereof is provided:
In another preferred embodiment, the compound of formula (I) or formula (XIV) is selected from the group consisting of
In the third aspect of the present invention, a pharmaceutical composition, where comprises the compound of the first aspect of the present invention, or the optical, tautomer, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and (2) pharmaceutically acceptable carriers is provided.
In the fourth aspect of the present invention, a use of the compound of the first aspect of the present invention, or an optical isomer, a pharmaceutically acceptable salt, a prodrug, a deuterated derivative, a hydrate, or a solvent compound thereof in the preparation of a pharmaceutical composition for the treatment of a disease, condition, or condition associated with KAT6 activity or expression.
In another preferred embodiment, said disease, condition or disease is selected from the group consisting of non-small cell lung cancer, small cell lung cancer, adenocarcinoma of the lung, squamous lung cancer, pancreatic cancer, colon cancer, thyroid cancer, embryonal rhabdomyosarcoma, granulosa cell tumor of the skin, melanoma, hepatocellular carcinoma, intrahepatic cholangiocarcinoma, rectal carcinoma, bladder carcinoma, pharyngolaryngeal cancer, breast carcinoma, vaginal carcinoma, prostate cancer, testicular cancer, brain tumor, glioblastoma, ovarian cancer, head and neck squamous cell carcinoma, cervical cancer, osteosarcoma, esophageal cancer, kidney cancer, skin cancer, gastric cancer, myeloid leukemia, lymphatic leukemia, myeloid fibrosis, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, monocytic leukemia, polycythemia megalosplenica, Eosinophilic leukocytosis syndrome multiple, bone marrow cancer, and various other solid and hematologic tumors, etc.
After long-term and intensive research, the present inventors have unexpectedly discovered a KAT6 protein kinase inhibitor of novel structure, as well as their preparation methods and applications. The compounds of the invention may be applied to the treatment of various diseases associated with the activity of acetyltransferase kinases. Based on the above findings, the inventors completed the present invention.
Unless otherwise stated, “or” as used herein has the same meaning as “and/or” (refers to “or” and “and”).
Unless otherwise specified, among all compounds of the present invention, each chiral carbon atom (chiral center) may optionally be in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.
As used herein, the term “alkyl”, alone or as part of another substituent, refers to a straight (ie, unbranched) or branched saturated hydrocarbon group containing only carbon atoms, or a combination of straight and branched chains. When the alkyl group has a carbon number limitation (e.g., C1-10), it means that the alkyl group has 1 to 10 carbon atoms. For example, C1-8 alkyl refers to an alkyl group containing from 1 to 8 carbon atoms, including methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or the like.
As used herein, the term “alkenyl”, when used alone or as part of another substituent, refers to a straight or branched, carbon chain group having at least one carbon-carbon double bond. Alkenyl groups can be substituted or unsubstituted. When the alkenyl group has a carbon number limit (e.g., C2-8), it means that the alkenyl group has 2-8 carbon atoms. For example, C2-8 alkenyl refers to alkenyl groups having 2-8 carbon atoms, including ethenyl, propenyl, 1,2-butenyl, 2,3-butenyl, butadienyl, or the like.
As used herein, the term “alkynyl”, when used alone or as part of another substituent, refers to an aliphatic hydrocarbon group having at least one carbon-carbon triple bond. The alkynyl group can be straight or branched, or a combination thereof. When the alkynyl group has a carbon number limitation (e.g., C2-8 alkynyl group), it means that the alkynyl group has 2 to 8 carbon atoms. For example, the term “C2-8 alkynyl” refers to a straight or branched alkynyl group having 2-8 carbon atoms, including ethynyl, propynyl, isopropynyl, butynyl, isobutynyl, secondary Butynyl, tert-butynyl, or the like.
As used herein, when used alone or as part of another substituent, the term “cycloalkyl” refers to a group having a saturated or partially saturated ring, a bicyclic or polycyclic (fused ring, bridged or spiro) ring system. When a certain cycloalkyl group has a carbon number limitation (e.g., C3-10), it means that the cycloalkyl group has 3 to 10 carbon atoms. In some preferred embodiments, the term “C3-8 cycloalkyl” refers to a saturated or partially saturated monocyclic or bicyclic alkyl group having from 3 to 8 carbon atoms, including cyclopropyl, cyclobutyl, cyclopentyl, cycloheptyl, or the like. “Spirocycloalkyl” refers to a bicyclic or polycyclic group that shares a carbon atom (called a spiro atom) between the monocyclic rings. These may contain one or more double bonds, but none of the rings have fully conjugated R electrons system. “Fused cycloalkyl” refers to an all-carbon bi-cyclic or polycyclic group in which each ring share two neighboring carbon atoms with other ring(s), which may contain one or more double bonds, but none of the rings have a fully conjugated π-electron system. “Bridge cycloalkyl” refers to an all-carbon polycyclic group in which two rings share two carbon atoms that are not directly bonded, which may contain one or more double bonds, but none of the rings have a fully conjugated π-electron system. The atoms contained in the cycloalkyl group are all carbon atoms. Some examples of cycloalkyl groups are as follows, and the present invention is not limited to the following cycloalkyl groups.
Unless otherwise stated, the following terms used in the specification and claims have the following meanings. “Aryl” means an all-carbon monocyclic or fused polycyclic (ie, a ring that shares a pair of adjacent carbon atoms) groups having a conjugated π-electron system, such as phenyl and naphthyl. The aryl ring may be fused to other cyclic groups (including saturated and unsaturated rings), but may not contain heteroatoms such as nitrogen, oxygen, or sulfur, while the point of attachment to the parent must be on the carbon atoms of the ring in a conjugated π-electron system. The aryl group can be substituted or unsubstituted. The following are some examples of aryl groups, and the present invention is not limited to the aryl groups described below.
“Heteroaryl” refers to an aromatic monocyclic or polycyclic group containing one to more heteroatoms (optionally from nitrogen, oxygen, and sulfur), or a polycyclic group formed by condensing a heterocyclic group (containing one to more heteroatoms, optionally selected from nitrogen, oxygen, and sulfur) with an aryl group, and the attachment site is located on the aryl group. The heteroaryl group can be optionally substituted or unsubstituted. The following are some examples of heteroaryl groups, and the present invention is not limited to the following heteroaryl groups.
“Heterocyclyl” means a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent wherein one or more of the ring atoms are selected from nitrogen, oxygen or sulfur and the remaining ring atoms are carbon. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl. Polycyclic heterocyclic group refers to a heterocyclic group including a spiro ring, a fused ring, and a bridged ring. “Spirocyclic heterocyclyl” refers to a polycyclic heterocyclic group in which each ring of the system shares an atom (referred to as a spiro atom) with other rings in the system, wherein one or more of the ring atoms is selected from the group consisting of nitrogen and oxygen. Or sulfur, the remaining ring atoms are carbon. “Fused ring heterocyclyl” refers to a polycyclic heterocyclic group in which each ring of the system shares an adjacent pair of atoms with other rings in the system, and one or more rings may contain one or more double bonds, but none One ring has a fully conjugated pi-electron system, and wherein one or more ring atoms are selected from nitrogen, oxygen or sulfur, and the remaining ring atoms are carbon. “Bridged heterocyclyl” refers to a polycyclic heterocyclic group in which any two rings share two atoms which are not directly bonded, these may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system, and wherein one or more of the ring atoms are selected from nitrogen, oxygen or sulfur, and the remaining ring atoms are carbon. If a heterocyclic group has both a saturated ring and an aromatic ring (for example, the saturated ring and the aromatic ring are fused together), the point attached to the parent must be on the saturated ring. Note: When the point attached to the parent is on the aromatic ring, it is called a heteroaryl group and is not called a heterocyclic group. Some examples of the heterocyclic group are as follows, and the present invention is not limited to the following heterocyclic group.
As used herein, the term “halogen”, when used alone or as part of another substituent, refers to F, Cl, Br, and I.
As used herein, the term “substituted” (when with or without “optionally”) means that one or more hydrogen atoms on a particular group are replaced by a particular substituent. Particular substituents are the substituents described above in the corresponding paragraphs, or the substituents which appear in the examples. Unless otherwise stated, an optionally substituted group may have a substituent selected from a particular group at any substitutable position of the group, and the substituents may be the same or different at each position. A cyclic substituent, such as a heterocyclic group, may be attached to another ring, such as a cycloalkyl group, to form a spirobicyclic ring system, i.e., the two rings have a common carbon atom. Those skilled in the art will appreciate that the combinations of substituents contemplated by the present invention are those that are stable or chemically achievable. The substituents are, for example but not limited to, C1-8 alkyl, C2-8 alkenyl, C2-8 alkynyl, C3-8 cycloalkyl, 3- to 12-membered heterocyclyl, aryl, heteroaryl, halogen, hydroxy, carboxy (—COOH), C1-8 aldehyde, C2-10 acyl, C2-10 ester, amino group.
For convenience and in accordance with conventional understanding, the term “optionally substituted” or “optionally substituted” applies only to sites which are capable of being substituted by a substituent, and does not include those which are not chemically achievable.
As used herein, unless otherwise specified, the term “pharmaceutically acceptable salt” refers to a salt that is suitable for contact with the tissue of a subject (eg, a human) without causing unpleasant side effects. In some embodiments, a pharmaceutically acceptable salt of a compound of the invention includes a salt (eg, a potassium salt, a sodium salt, a magnesium salt, a calcium salt) of a compound of the invention having an acidic group or is basic A salt of a compound of the invention (e.g., a sulfate, a hydrochloride, a phosphate, a nitrate, a carbonate).
The present invention provides a class of compounds of formula (I), or their deuterated derivatives, their pharmaceutically acceptable salts, optical isomers (enantiomers or diastereomers, if any case), hydrates, solvates, or pharmaceutical combinations comprising the compound represented by formula (I), its optical isomers, pharmaceutically acceptable salts, prodrugs, deuterated forms, hydrates, and solvates for inhibiting KAT6 kinase activity.
The compound of the present invention can be used as KAT6 kinase inhibitor.
The present invention is a single inhibitor of KAT6 for the purpose of preventing, alleviating or curing a disease by modulating the activity of KAT6. The referred diseases include, but are not limited to: non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, squamous lung cancer, pancreatic cancer, colon cancer, thyroid cancer, embryonal rhabdomyosarcoma, granular cell tumor of the skin, melanoma, hepatocellular carcinoma, intrahepatic cholangiocarcinoma, rectal carcinoma, bladder cancer, pharyngeal carcinoma, breast carcinoma, vaginal carcinoma, prostate carcinoma, testicular carcinoma, brain tumor, glioma, ovarian carcinoma, squamous cancer of the head and neck, Cervical Cancer, Osteosarcoma, Esophageal Cancer, Kidney Cancer, Skin Cancer, Gastric Cancer, Myeloid Leukemia, Lymphoid Leukemia, Myelofibrosis, B-cell Lymphoma, T-cell Lymphoma, Hodgkin's Lymphoma, Non-Hodgkin's Lymphoma, Mononuclear Cell Leukemia, Erythropoietic Eosinophilic Leukocytosis, Eosinophilic Leukocytosis Syndrome Multi-Cellular, Myeloid Carcinoma and other solid and hematologic tumors.
The compounds of the present invention and their deuterated forms, as well as pharmaceutically acceptable salts or isomers (if present) or hydrates and/or compositions thereof can be combined with pharmaceutically acceptable excipients or carriers formulated together, the resulting composition can be administered to humans or animals for the treatment of disorders, symptoms and diseases. The composition can be: tablets, pills, suspensions, solutions, emulsions, capsules, aerosols, sterile injections, sterile powders and the like. In a preferred embodiment, the pharmaceutical composition is a dosage form suitable for oral administration, including but not limited to tablets, solutions, suspensions, capsules, granules, and powders. The amount of the compound or pharmaceutical composition administered to the patient is not fixed, and is usually administered in a pharmaceutically effective amount. At the same time, the amount of the compound actually administered can be determined by the physician according to the actual situation, including the disease to be treated, the route of administration selected, the actual compound administered, the individual condition of the patient, and so on. The dosage of the compound of the present invention depends on the specific use of the treatment, the mode of administration, the state of the patient, and the judgment of the physician. The ratio or concentration of the compound of the present invention in the pharmaceutical composition depends on a variety of factors, including dosage, physical and chemical properties, route of administration, and the like.
It is to be understood that within the scope of the present invention, the various technical features of the present invention and the various technical features specifically described hereinafter (as in the embodiments) may be combined with each other to form a new or preferred technical solution.
Since the compound of the present invention has excellent inhibitory activity against KAT6, the compound of the present invention and its various crystal forms, optical isomers, pharmaceutically acceptable inorganic or organic salts, prodrugs, deuterated forms, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredients can be used to treat, prevent and alleviate diseases associate with the activity or expression of KAT6.
The pharmaceutical compositions of the present invention comprise a safe or effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier. By “safe and effective amount” it is meant that the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. In general, the pharmaceutical compositions contain from 1 to 2000 mg of the compound of the invention per agent, more preferably from 5 to 200 mg of the compound of the invention per agent. Preferably, the “one dose” is a capsule or tablet.
“Pharmaceutically acceptable carrier” means: one or more compatible solid or liquid fillers or gel materials which are suitable for human use and which must be of sufficient purity and of sufficiently low toxicity. By “compatibility” it is meant herein that the components of the composition are capable of intermingling with the compounds of the invention and with each other without significantly reducing the efficacy of the compound. Examples of pharmaceutically acceptable carriers include, but are not limited to, filler (or diluent), disintegrant, lubricant, binder, matrix, emulsifiers, run wet agents, colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.
The mode of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.
Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier). In capsules, tablets and pills, the dosage form may also contain a buffer.
Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other materials known in the art. They may contain opacifying agents and the release of the active compound or compound in such compositions may be released in a portion of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric and waxy materials. If necessary, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.
In addition to these inert diluents, the compositions may contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.
In addition to the active compound, the suspension may contain suspending agents.
Compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion. Suitable aqueous and nonaqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.
Dosage forms for the compounds of the invention for topical administration include ointments, powders, patches, propellants and inhalants. The active ingredient is admixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or, if necessary, propellants.
The compounds of the invention may be administered alone or in combination with other pharmaceutically acceptable compounds.
When a pharmaceutical composition is used, a safe and effective amount of a compound of the invention is administered to a mammal (e.g., a human) in need of treatment wherein the dosage is a pharmaceutically effective dosage, for a 60 kg body weight, The dose to be administered is usually from 1 to 2000 mg, preferably from 5 to 500 mg. Of course, specific doses should also consider factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled physician.
The main advantages of the present invention include:
The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually in accordance with conventional conditions or according to the conditions recommended by the manufacturer. Percentages and parts are by weight unless otherwise stated.
Some of the representative compounds of the present invention can be prepared by the following synthetic methods, and in each of the following reaction formulas, the reagents and conditions for each step can be selected from those conventional in the field for carrying out such preparation methods, and the above selections can be made by a person skilled in the art according to the knowledge in the field after the structures of the compounds of the present invention are disclosed.
Some compounds of the present invention can be prepared by the following methods:
Compound (VIIIa) can be synthesized by using the method described in patent WO2020254946, while compound (VIIIb) can be synthesized using relevant references. In an inert solvent, compound (VIIIa) reacts with compound (VIIIb) to obtain compound (VIII). The definition of A in the above reaction formula is the same as the description in claim 1.
The compound of formula (II) of the present invention can be prepared by the following methods:
Compound (IIa) can be synthesized using the method described in patent WO2020254946. In an inert solvent, compound (IIa) is reacted with compound (IIb) to obtain compound (IIc), which is then combined with the corresponding intermediate or reagent through a metal catalyzed coupling reaction or substitution reaction to synthesize the target compound (II). The definitions of R2, R3, R7, R8, R9, R10, n, m, h, j, D and E in the above reaction formulas are the same as those in the claims.
The compound of formula (III) of the present invention can be prepared by the following methods:
In an inert solvent, the target compound (IIIb) is synthesized by metal catalyzed coupling reaction or substitution reaction between compound (IIc) and compound (IIIa). Compound (III) is obtained through wittig reaction with compound (IIIb). The definitions of R2, R7, R8, R9, R10, R12, R13, R14, p, q, h, j, m, t, D and U in the above reaction formulas are the same as those in the claims.
The compound of formula (IV) of the present invention can be prepared by the following methods:
Compound (IV) is obtained by metal catalyzed coupling reaction between compound (IIc) and compound (IVa) in an inert solvent. The definitions of R2, R4, R7, R8, R9, R10, h, j, and m in the above reaction formulas are the same as those in the claims.
The compound of formula (V) of the present invention can be prepared by the following methods:
In an inert solvent, compound (Va) reacts with chlorosulfonic acid to obtain compound (Vb). Compound (Vb) reacts with compound (IIa) to obtain compound (Vc), which undergoes a wittig reaction to obtain compound (V). The definitions of R2, R6, R7, R8, R9, R10, Rh, Ri, a, b, f, g, h, and j in the above reaction formulas are the same as those in the claims.
The compound of formula (VI) of the present invention can be prepared by the following methods:
In an inert solvent, compound (Vb) reacts with compound (VIa) to obtain compound (VIb), which is then subjected to a Wittig reaction to obtain compound (VIc). The protective group is removed to obtain compound (VId). The compound (VIe) was synthesized using the method described in patent WO2020216701. Compound (VId) reacts with compound (VIe) to obtain compound (VI). The definitions of R2, R6, R11, Rh, Ri, X, a, b, g, f, and k in the above reaction formulas are the same as those in the claims.
The compound of formula (VII) of the present invention can be prepared by the following method:
In an inert solvent, compound (IIb) is reacted with compound (VIa) to obtain compound (VIIa), which is then subjected to a metal catalyzed coupling or substitution reaction to obtain compound (VIIb). Compound (VIIb) is then subjected to a Wittig reaction to obtain compound (VIIc), and the protective group is removed to obtain compound (VIId). The compound (VIe) was synthesized using the method described in patent WO2020216701. Compound (VIId) reacts with compound (VIe) to obtain compound (VII). The definitions of R2, R11, R12, R13, R14, X, D, U, p, q, t, m, and k in the above reaction formulas are the same as those in the claims.
The compound 1a was synthesized according to WO20130158004. The compound 1a (205 mg, 0.80 mmol) was dissolved in THF (Tetrahydrofuran, 10 mL). The mixture was cooled to −78° C. and nBuLi (2.5 M in hexane, 0.35 mL) was added. The reaction mixture was stirred at this temperature for 1 h. Then Bis (sulfur dioxide)-1,4-diazabicyclo [2.2.2]octane adduct (115 mg, 0.48 mmol) was added. The mixture was warmed to RT (room temperature) and stirred for 1 h. The reaction mixture was concentrated. The crude was washed by methyl tert-butyl ether (MTBE) and filterated to give solid compound 1b (180 mg, 91%). The product was used to the next step directly.
The compound 1b (203 mg, 0.82 mmol) was dissolved in dichloromethane (DCM, 5 mL). Then Chlorobutyrylimide (109 mg, 0.82 mmol) was added under ice bath. The reaction mixture was stirred at this temperature for 1 h. The reaction mixture was filterated and concentrated under reduced pressure. The crude was purified by column chromatography to give white solid compound 1c (94 mg, yield 42%). 1H NMR (500 MHz, CDCl3) δ=7.78 (d, J=9.0 Hz, 1H), 6.41 (d, J=2.0 Hz, 1H), 6.36 (dd, J=9.0 Hz, J=2.0 Hz, 1H), 4.68-4.62 (m, 1H), 2.44-2.38 (m, 2H), 2.17-2.10 (m, 2H), 1.89-1.80 (dd, J=21.0, 10.4, 1H), 1.72-1.62 (m, 1H) ppm
The compound 1d was synthesized according to WO2020254946. The compound 1c (30 mg, 0.11 mmol) was dissolved in pyrdine (0.2 mL). Then 1d (13 mg, 0.05 mmol) was added. The reaction mixture was stirred at 120° C. for 2 h in a sealed tube. The reaction was concentrated under reduced pressure. The crude was purified by Pre-TLC (Preparative TLC) to give white solid compound 1 (5 mg, yield 20%). 1H NMR (500 MHz, CDCl3) δ 8.05-7.94 (m, 2H), 7.60 (s, 1H), 7.46 (s, 1H), 6.79 (s, 1H), 6.50 (s, 1H), 6.42 (dd, J=8.8, 1.8 Hz, 1H), 6.37-6.31 (m, 2H), 5.41 (s, 2H), 4.69-4.61 (m, 1H), 3.96 (s, 3H), 3.84 (s, 3H), 2.48-2.38 (m, 2H), 2.19-2.08 (m, 2H), 1.91-1.83 (m, 1H), 1.74-1.65 (m, 1H). MS m z 485.4 M+H]+
The P-Methoxybromobenzene 2a (1.0 g, 5.38 mmol) was dissolved in dichloromethane (DCM, 5 mL). Then Chlorosulfonic acid (1.0 mL) was added under ice bath. The reaction was stirred at RT for 3 h. The reaction was poured into ice water and extracted with DCM (20 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by column chromatography to give white solid compound 2b (1.4 g, yield 92%).
The compound 1d was synthesized according to WO2020254946. The compound 2b (70 mg, 0.25 mmol) was dissolved in pyridine (0.2 mL). Then 1d (30 mg, 0.12 mmol) was added. The reaction mixture was stirred at 120° C. for 2 h in a sealed tube. The reaction was concentrated under reduced pressure. The crude was purified by column chromatography to give white solid compound 2c (30 mg, yield 50%). MS m/z 493.3 [M+H]+.
The compound 2c (25 mg, 0.05 mmol), Methanesulfonic acid (2-dicyclohexylphosphine-2′, 6′-diisopropyloxy-1,1′-biphenyl) (2-amino-1,1′-biphenyl-2-yl) palladium (II) (4 mg, 0.005 mmol), 2-Dicyclohexylphosphate-20′,6′-diisopropyloxy-1,1′-biphenyl (5 mg, 0.01 mmol), sodium tert-butoxide (19 mg, 0.20 mmol) and azetidine hydrochloride (14 mg, 0.15 mmol) were dissolved in dioxane (0.5 mL) The reaction mixture was stirred at 90° C. for 3 h in a sealed tube under N2. TLC showed the reaction was completed. The mixture was filterated through diatomite and concentrated under reduced pressure. The crude was purified by Pre-TLC to give white solid compound 2 (3 mg, yield 13%). 1H NMR (500 MHz, CDCl3) δ 8.08 (s, 1H), 7.56 (d, J=1.2 Hz, 1H), 7.43 (d, J=2.0 Hz, 1H), 7.29 (br, 1H), 6.84 (d, J=8.8 Hz, 1H), 6.78 (s, 1H), 6.73 (br, 1H), 6.44 (s, 1H), 6.31 (t, J=1.9 Hz, 1H), 5.37 (s, 2H), 3.95 (s, 3H), 3.95-3.91 (m, 4H), 3.83 (s, 3H), 2.43-2.36 (m, 2H) ppm. MS m/z 470.4 [M+H]+.
The compound 2c (28 mg, 0.06 mmol), Trimethylsilyl acetylene (11 mg, 0.11 mmol), Pd(dppf)Cl2 (9 mg, 0.01 mmol), Tri-tert-butylphosphine (6 mg, 0.03 mmol) and Cuprous iodide (CuI, 2 mg, 0.01 mmol) were dissolved in Tetrahydrofuran/triethylamine (0.5 mL/0.5 mL) The reaction mixture was stirred at 90° C. for 3 h in a sealed tube under N2. The mixture was filterated through diatomite and concentrated under reduced pressure. The crude was purified by column chromatography to give white solid compound 3a (9 mg, yield 31%). MS m/z 511.4 [M+H]+.
The compound 3a (9 mg, 0.02 mmol) was dissolved in acetonitrile (0.1 mL). Then potassium carbonate (5 mg, 0.04 mmol) was added. The reaction mixture was stirred at RT for 2 h. The mixture was filterated through and concentrated under reduced pressure. The crude was purified by Pre-TLC to give white solid compound 3 (4 mg, yield 49%). 1H NMR (500 MHz, CDCl3) δ 8.27 (d, J=2.1 Hz, 1H), 7.63 (dd, J=8.6 Hz, J=2.1 Hz, 1H), 7.57 (d, J=1.6 Hz, 1H), 7.44 (d, J=2.2 Hz, 1H), 6.90 (d, J=8.6 Hz, 1H), 6.79 (s, 1H), 6.45 (s, 1H), 6.32 (t, J=2.1 Hz, 1H), 5.38 (s, 2H), 3.96 (s, 3H), 3.92 (s, 3H), 3.07 (s, 1H) ppm. MS m/z 439.3 [M+H]+.
The compound 4a (5.0 g, 23.03 mmol) was dissolved in dichloromethane (20 mL). Then Chlorosulfonic acid (3.0 mL) was added under ice bath. The reaction was stirred at RT for 1 h. The reaction was poured into ice water and extracted with DCM (20 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by column chromatography to give white solid compound 4b (500 mg). 1HNMR (500 MHz, CDCl3) δ 6.82 (s, 2H), 3.98 (s, 6H)
The compound 4b (150 mg, 0.48 mmol) was dissolved in pyrdine (0.2 mL). Then 1d (58 mg, 0.24 mmol) was added. The reaction was stirred at 120° C. for 2 h in a sealed tube.
The reaction was concentrated under reduced pressure. The crude was purified by column chromatography to give white solid compound 4c (55 mg, yield 44%). MS m/z 523.0 [M+H]+.
The compound 4c (35 mg, 0.07 mmol), triethylamine (0.2 mL), di (triphenylphosphine) palladium dichloride (5 mg, 0.007 mmol) and Trimethylsilyl acetylene (11 mg, 0.11 mmol) were dissolved in dimethyl sulfoxide (DMSO, 1 mL) The reaction was stirred at 100° C. for 2 h in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (EA, 5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by column chromatography to give white solid compound 4d (27 mg, yield 75%). MS m/z 541.2 [M+H]+.
The compound 4d (27 mg, 0.05 mmol) was dissolved in MeOH (methanol, 0.2 mL) and N, N-dimethylformamide (DMF, 0.2 mL). Then potassium carbonate (35 mg, 0.25 mmol) was added. The reaction mixture was stirred at RT for 2 h. The mixture was filterated and concentrated under reduced pressure. The crude was purified by Pre-TLC to give white solid compound 3 (14 mg, yield 61%). MS m/z 469.0 [M+H]+. 1H NMR (500 MHz, CDCl3-d) δ 8.18 (s, 1H), 7.58 (d, J=1.9 Hz, 1H), 7.45 (d, J=2.3 Hz, 1H), 6.79 (s, 1H), 6.68 (s, 2H), 6.45 (s, 1H), 6.33 (t, J=2.1 Hz, 1H), 5.39 (s, 2H), 3.96 (s, 3H), 3.87 (s, 6H), 3.21 (s, 1H) ppm.
The compound 2c (20 mg, 0.04 mmol), sodium tert-butoxide (12 mg, 0.12 mmol), Methanesulfonic acid (2-dicyclohexylphosphine-2′, 6′-diisopropyloxy-1,1′-biphenyl) (2-amino-1,1′-biphenyl-2-yl) palladium (II) (2.6 mg, 0.003 mmol), 2-Dicyclohexylphosphate-20′,6′-diisopropyloxy-1,1′-biphenyl (2.6 mg, 0.006 mmol), and methylpiperazine (12 mg, 0.12 mmol) were dissolved in DMF (1 mL). The reaction mixture was stirred at 85° C. for 3 h in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by Pre-TLC to give white solid compound 5 (8 mg, yield 38%). MS m/z 513.2 [M+H]+. 1H NMR (500 MHz, DMSO-d6) δ 9.90 (s, 1H), 7.86 (d, J=2.1 Hz, 1H), 7.49 (d, J=2.0 Hz, 1H), 7.39 (d, J=3.0 Hz, 1H), 7.11 (dd, J=9.0, 3.0 Hz, 1H), 7.00 (d, J=9.0 Hz, 1H), 6.73 (s, 1H), 6.65 (s, 1H), 6.29 (t, J=2.0 Hz, 1H), 5.41 (s, 2H), 3.83 (s, 3H), 3.68 (s, 3H), 3.09 (m, 4H), 2.74 (m, 4H), 2.42 (s, 3H).
The compound 2c (20 mg, 0.04 mmol), cesium carbonate (40 mg, 0.12 mmol), Methanesulfonic acid (2-dicyclohexylphosphine-2′, 6′-diisopropyloxy-1,1′-biphenyl) (2-amino-1,1′-biphenyl-2-yl) palladium (II) (2.6 mg, 0.003 mmol), 2-Dicyclohexylphosphate-20′,6′-diisopropyloxy-1,1′-biphenyl (2.6 mg, 0.006 mmol), and Morpholine (11 mg, 0.12 mmol) were dissolved in DMF (1 mL). The reaction mixture was stirred at 100° C. for 3 h in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by Pre-TLC to give white solid compound 6 (4 mg, yield 20%). MS m z 500.1 [M+H]+. 1H NMR (500 MHz, DMSO-d6) δ 10.05 (s, 1H), 7.87 (d, J=2.2 Hz, 1H), 7.49 (d, J=2.2 Hz, 1H), 7.33 (d, J=3.0 Hz, 1H), 7.15 (m, 2H), 6.78 (m, 2H), 6.30 (t, J=2.0 Hz, 1H), 5.43 (s, 2H), 3.83 (s, 3H), 3.73-3.66 (m, 7H), 3.10-2.90 (m, 4H).
The compound 2c (20 mg, 0.04 mmol), cesium carbonate (40 mg, 0.12 mmol), di (triphenylphosphine) palladium dichloride (2.8 mg, 0.004 mmol) and Cyclopropyl Acetylene (8 mg, 0.12 mmol) were dissolved in DMF (1 mL) The reaction mixture was stirred at 100° C. for 2 h in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (EA, 5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by column chromatography to give white solid compound 7 (5 mg, yield 26%). MS m z 479.1 [M+H]+. 1H NMR (500 MHz, DMSO-d6) δ 10.46 (s, 1H), 7.88 (d, J=2.0 Hz, 1H), 7.70 (d, J=2.0 Hz, 1H), 7.64-7.58 (m, 1H), 7.50 (d, J=1.3 Hz, 1H), 7.17 (d, J=8.7 Hz, 1H), 6.85 (s, 1H), 6.75 (s, 1H), 6.31-6.29 (m, 1H), 5.44 (s, 2H), 3.81 (s, 3H), 3.78 (s, 3H), 1.54-1.49 (m, 1H), 0.89-0.83 (m, 2H), 0.73-0.75 (m, 2H).
The compound 2b (140 mg, 0.49 mmol) was dissolved in Tetrahydrofuran (5 mL). Then ammonia methanol solution (7 M, 1.0 mL, 7 mmol) was added. The reaction was stirred at RT for 1 h in a sealed tube. The reaction was concentrated under reduced pressure. The crude was purified by column chromatography to give white solid compound 8a (100 mg, yield 77%).
The compound 8b was synthesized according to WO2020216701. The compound 8a (50 mg, 0.19 mmol), 8b (47 mg, 0.21 mmol) and N,N-diisopropylethylamine (107 mg, 0.76 mmol) was dissolved in DCM (2 mL). Then 1H-Benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (130 mg, 0.23 mmol) was added. The reaction was stirred at RT for overnight. The mixture was quenched with water and extracted with ethyl acetate (EA, 5 mL×3). The collected organic phase was washed with sat·NaCl (sat.: saturated), dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by column chromatography to give white solid compound 8c (50 mg, yield 59%). MS m/z 453.0 [M+H]+
The compound 8c (25 mg, 0.06 mmol), cesium carbonate (54 mg, 0.17 mmol), di (triphenylphosphine) palladium dichloride (3.9 mg, 0.006 mmol) and Cyclopropyl Acetylene (11 mg, 0.17 mmol) were dissolved in DMF (1 mL) The reaction mixture was stirred at 100° C. for 2 h in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (EA, 5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by Pre-TLC to give white solid compound 8 (11 mg, yield 46%). MS m/z 439.0 [M+H]+. 1H NMR (500 MHz, DMSO-d6) δ 12.55 (s, 1H), 7.91 (br, 1H), 7.80 (d, J=2.1 Hz, 1H), 7.63 (s, 1H), 7.55 (d, J=8.8 Hz, 1H), 7.18 (d, J=7.8 Hz, 1H), 6.85 (d, J=8.8 Hz, 1H), 6.77 (s, 1H), 3.86 (s, 3H), 2.98 (s, 6H), 1.58-1.52 (m, 1H), 0.90-0.87 (m, 2H), 0.80-0.74 (m, 2H).
The compound 4c (80 mg, 0.15 mmol), propyne (1 M, 0.45 mmol), cesium carbonate (149 mg, 0.46 mmol) and di (triphenylphosphine) palladium dichloride (14 mg, 0.02 mmol) were dissolved in DMF (1 mL) The reaction mixture was stirred at 100° C. for 3 h i under N2. The mixture was filterated through diatomite and concentrated under reduced pressure. The crude was purified by Pre-TLC (MeOH:DCM=1:20, V:V) to give compound 9 (7 mg, yield 9%). 1H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.88 (d, J=2.1 Hz, 1H), 7.50 (d, J=1.4 Hz, 1H), 6.90-6.70 (m, 4H), 6.30 (t, J=2.0 Hz, 1H), 5.44 (s, 2H), 3.84 (s, 3H), 3.74 (s, 6H), 2.07 (s, 3H) ppm. MS m/z 483.0 [M+H]+.
The compound 4c (50 mg, 0.10 mmol), Cyclopropyl Acetylene (13 mg, 0.19 mmol), cesium carbonate (93 mg, 0.29 mmol) and di (triphenylphosphine) palladium dichloride (13 mg, 0.02 mmol) were dissolved in DMF (1 mL). The reaction mixture was stirred at 100° C. for 3 h under N2. The mixture was filterated through diatomite and concentrated under reduced pressure. The crude product was purified by Pre-TLC (MeOH:DCM=1:20, V:V) to give compound 10 (12 mg, yield 25%). 1H NMR (500 MHz, DMSO-d6) δ 9.79 (brs, 1H), 7.88 (d, J=2.1 Hz, 1H), 7.50 (d, J=1.4 Hz, 1H), 6.82 (s, 1H), 6.77-6.68 (m, 3H), 6.30 (t, J=2.0 Hz, 1H), 5.44 (s, 2H), 3.85 (s, 3H), 3.74 (s, 6H), 1.60-1.53 (m, 1H), 0.95-0.89 (m, 2H), 0.80-0.73 (m, 2H) ppm. MS m/z 508.9 [M+H]+.
The compound 11a was synthesized according to WO2006610629. The compound 11a (300 mg, 1.05 mmol) was dissolved in pyrdine (1.2 mL). Then 1d (128 mg, 0.53 mmol) was added. The reaction mixture was stirred at 120° C. for 2 h in a sealed tube. The reaction mixture was concentrated under reduced pressure. The crude was purified by column chromatography to give white solid compound 11b (100 mg, yield 39%). MS m/z 494.9 [M+H]+.
The compound 11b (18 mg, 0.04 mmol), triethylamine (0.2 mL), di (triphenylphosphine) palladium dichloride (2 mg, 0.003 mmol) and Trimethylsilyl acetylene (12 mg, 0.12 mmol) were dissolved in dimethyl sulfoxide (1 mL) The reaction mixture was stirred at 100° C. for 2 h in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (EA, 5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by column chromatography to give white solid product 11c (5 mg, yield 27%). MS m/z 511.1 [M+H]+.
The compound 11c (5 mg, 0.01 mmol) was dissolved in MeOH (1 mL) and N,N-dimethylformamide (DMF, 0.2 mL). Then potassium carbonate (4 mg, 0.03 mmol) was added. The reaction was stirred at RT for 2 h. The mixture was filterated through and concentrated under reduced pressure. The crude was purified by Pre-TLC (MeOH:DCM=1:20, V:V) to give white solid compound 11 (2 mg, yield 47%). 1H NMR (500 MHz, CDCl3) δ 8.08 (d, J=8.1 Hz, 1H), 8.02 (s, 1H), 7.57 (d, J=1.2 Hz, 1H), 7.43 (d, J=2.0 Hz, 1H), 7.20 (d, J=8.1 Hz, 1H), 7.03 (s, 1H), 6.79 (s, 1H), 6.45 (s, 1H), 6.32 (t, J=2.0 Hz, 1H), 5.38 (s, 2H), 3.96 (s, 3H), 3.91 (s, 3H), 3.24 (s, 1H) ppm. MS m/z 439.0 [M+H]+.
The compound 11b (18 mg, 0.04 mmol), cesium carbonate (40 mg, 0.12 mmol), di (triphenylphosphine) palladium dichloride (2.8 mg, 0.004 mmol) and Cyclopropyl Acetylene (8 mg, 0.12 mmol) were dissolved in DMF (1 mL). The reaction mixture was stirred at 100° C. for 2 h under N2. The mixture was quenched with water and extracted with ethyl acetate (5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by Pre-TLC (MeOH:DCM=1:20, V:V) to give white solid compound 12 (10 mg, yield 57%). 1H NMR (500 MHz, DMSO-d6) δ 10.34 (s, 1H), 7.87 (d, J=2.1 Hz, 1H), 7.71 (d, J=8.1 Hz, 1H), 7.50 (d, J=1.6 Hz, 1H), 7.13 (s, 1H), 7.05 (d, J=8.1 Hz, 1H), 6.83 (s, 1H), 6.73 (s, 1H), 6.30 (t, J=2.1 Hz, 1H), 5.44 (s, 2H), 3.81 (s, 3H), 3.76 (s, 3H), 1.58 (m, 1H), 0.97-0.87 (m, 2H), 0.80-0.70 (m, 2H) ppm. MS m/z 479.1 [M+H]+.
The compound 2c (30 mg, 0.06 mmol), propyne in THF (1 M, 0.30 mmol), cesium carbonate (60 mg, 0.18 mmol) and di (triphenylphosphine) palladium dichloride (8 mg, 0.01 mmol) were dissolved in DMF (1 mL) The reaction was stirred at 100° C. for 3 h in a sealed tube under N2. The reaction mixture was filterated through diatomite and concentrated under reduced pressure. The crude was purified by Pre-TLC (MeOH:DCM=1:20, V:V) to give compound 13 (1.6 mg, yield 6%). 1H NMR (500 MHz, DMSO-d6) δ 10.50 (s, 1H), 7.87 (d, J=1.6 Hz, 1H), 7.73 (br, 1H), 7.62 (br, 1H), 7.49 (s, 1H), 7.18 (br, 1H), 6.84 (br, 1H), 6.74 (br, 1H), 6.30 (s, 1H), 5.44 (s, 2H), 3.81 (s, 3H), 3.78 (s, 3H), 2.02 (s, 3H). MS m/z 453.0 [M+H]+.
The compound 4c (60 mg, 0.11 mmol), 3-ethynyloxycyclohexane (19 mg, 0.23 mmol), cesium carbonate (112 mg, 0.34 mmol) and di (triphenylphosphine) palladium dichloride (13 mg, 0.12 mmol) were dissolved in DMF (1 mL) The reaction mixture was stirred at 100° C. for 2 h in a sealed tube under N2. The mixture was filterated through diatomite and concentrated under reduced pressure. The crude was purified by Pre-TLC (MeOH:DCM=1:20) to give compound 14 (13 mg, yield 22%). 1H NMR (500 MHz, DMSO-d6) δ 9.88 (s, 1H), 7.88 (d, J=2.0 Hz, 1H), 7.50 (d, J=1.4 Hz, 1H), 6.86-6.80 (m, 3H), 6.76 (s, 1H), 6.30 (t, J=2.0 Hz, 1H), 5.45 (s, 2H), 4.80 (dd, J=8.5, 5.5 Hz, 2H), 4.62 (dd, J=7.0, 5.5 Hz, 2H), 4.22-4.14 (m, 1H), 3.85 (s, 3H), 3.77 (s, 6H) ppm. MS m/z 525.0 [M+H]+.
The compound 4c (60 mg, 0.11 mmol), 15a (40 mg, 0.34 mmol), cesium carbonate (112 mg, 0.34 mmol) and di(triphenylphosphine) palladium dichloride (16 mg, 0.02 mmol) were dissolved in DMF (1 mL). The reaction mixture was stirred at 100° C. for 2 h under N2. The mixture was filterated through diatomite and concentrated under reduced pressure. The crude was purified by Pre-TLC (MeOH:DCM=1:20, V:V) to give compound 15 (14 mg, yield 22%). 1H NMR (500 MHz, DMSO-d6) δ 9.90 (s, 1H), 7.89 (d, J=2.1 Hz, 1H), 7.50 (d, J=1.3 Hz, 1H), 6.83 (s, 1H), 6.80 (s, 2H), 6.76 (s, 1H), 6.31 (t, J=2.0 Hz, 1H), 5.45 (s, 2H), 3.85 (s, 3H), 3.76 (s, 6H), 3.32-3.25 (m, 1H), 3.10-3.00 (m, 2H), 2.81-2.69 (m, 2H) ppm. MS m/z 559.0 [M+H]+.
The compound 11b (28 mg, 0.06 mmol), cesium carbonate (59 mg, 0.18 mmol) and di (triphenylphosphine) palladium dichloride (2 mg, 0.003 mmol) and propyne (24 mg, 0.6 mmol) were dissolved in DMF (1 mL) The reaction mixture was stirred at 100° C. for 3 h in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by Pre-TLC (MeOH:DCM=1:20, V:V) to give white solid compound 16 (22 mg, yield 86%). 1H NMR (500 MHz, CDCl3) δ 8.06-7.97 (m, 2H), 7.57 (d, J=1.6 Hz, 1H), 7.43 (d, J=2.1 Hz, 1H), 7.08 (dd, J=8.1, 1.2 Hz, 1H), 6.92 (d, J=1.0 Hz, 1H), 6.79 (s, 1H), 6.44 (s, 1H), 6.32 (t, J=2.1 Hz, 1H), 5.38 (s, 2H), 3.96 (s, 3H), 3.88 (s, 3H), 2.05 (s, 3H) ppm. MS m/z 453.0 [M+H]+.
The compound 11b (20 mg, 0.04 mmol), sodium tert-butoxide (31 mg, 0.32 mmol), Methanesulfonic acid (2-dicyclohexylphosphine-2′, 6′-diisopropyloxy-1,1′-biphenyl) (2-amino-1,1′-biphenyl-2-yl) palladium (II) (2.6 mg, 0.006 mmol), 2-Dicyclohexylphosphate-20′,6′-diisopropyloxy-1,1′-biphenyl (5 mg, 0.01 mmol) and 2d (19 mg, 0.2 mmol) was dissolved in DMF (1 mL). The reaction mixture was stirred at 90° C. for 2 h in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by Pre-TLC (MeOH:DCM=1:20, V:V) to give white solid compound 17 (8 mg, yield 42%). 1H NMR (500 MHz, CDCl3) δ 7.97 (s, 1H), 7.88 (d, J=8.7 Hz, 1H), 7.61 (s, 1H), 7.50 (s, 1H), 6.77 (s, 1H), 6.50 (s, 1H), 6.36 (s, 1H), 5.97 (dd, J=8.7, 1.9 Hz, 1H), 5.71 (d, J=1.9 Hz, 1H), 5.42 (s, 2H), 3.99-3.92 (m, 7H), 3.83 (s, 3H), 2.47-2.35 (m, 2H) ppm. MS m/z 470.0 [M+H]+.
The compound 18a (1.6 g, 5.11 mmol), potassium thioacetate (875 mg, 7.66 mmol), 1,10-Phenanthroline (460 mg, 2.56 mmol), and cuprous iodide (460 mg, 2.56 mmol) was dissolved in toluene (15 mL). The reaction mixture was stirred at 100° C. for 20 h. The mixture was quenched with water and extracted with ethyl acetate (20 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by column chromatography to give reddish brown solid compound 18b (1.0 g, yield 75%).
The N-chlorosuccinimide (2.3 g, 17.2 mmol) was dissolved in hydrochloric acid aqueous solution (2 M, 2 mL) and MeCN (10 mL). Then 18b (1.1 g, 4.2 mmol) in MeCN (2 mL) was added dropwise at RT. The reaction mixture was stirred at RT for 1 h. The mixture was quenched with water and extracted with ethyl acetate (20 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by column chromatography to give reddish brown solid compound 18c (1.0 g, yield 83%). 1H NMR (500 MHz, CDCl3) δ 7.47-7.39 (m, 2H), 7.10 (dd, J=7.5, 2.1 Hz, 1H), 4.05 (s, 3H).
The compound 18c (117 mg, 0.41 mmol) was dissolved in pyrdine (0.2 mL). Then 1d (50 mg, 0.20 mmol) was added. The reaction mixture was stirred at 120° C. for 1 h in a sealed tube. The reaction was concentrated under reduced pressure. The crude was purified by column chromatography to give white solid compound 18d (50 mg, yield 50%). MS m z 492.8 [M+H]+.
The compound 18d (25 mg, 0.05 mmol), triethylamine (0.2 mL), di (triphenylphosphine) palladium dichloride (5 mg, 0.007 mmol) and Triisopropylsilyacetylene (27 mg, 0.15 mmol) were dissolved in dimethyl sulfoxide (1 mL) The reaction was stirred at 100° C. for overnight in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (EA, 5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by column chromatography to give white solid compound 18e (25 mg, yield 83%). MS m/z 595.1 [M+H]+.
The compound 18e (10 mg, 0.017 mmol) was dissolved in DMF (1 mL). Then Cesium Fluoride (15 mg, 0.1 mmol) was added. The reaction mixture was stirred at RT for 5 h. The mixture was filterated and concentrated under reduced pressure. The crude was purified by Pre-TLC (MeOH:DCM=1:20) to give white solid compound 18 (3 mg, yield 40%). MS m/z 439.0 [M+H]+. 1H NMR (500 MHz, CDCl3) δ 8.18 (s, 1H), 7.67 (s, 1H), 7.53 (s, 1H), 7.42 (t, J=8.1 Hz, 1H), 7.29 (d, J=7.7 Hz, 1H), 6.96 (d, J=8.4 Hz, 1H), 6.83 (s, 1H), 6.66 (s, 1H), 6.40 (s, 1H), 5.49 (s, 2H), 3.99 (s, 3H), 3.91 (s, 3H), 3.58 (s, 1H).
The compound 4c (25 mg, 0.05 mmol), cesium carbonate (46 mg, 0.14 mmol) and di (triphenylphosphine) palladium dichloride (2.8 mg, 0.004 mmol) and 3-alkynylpyridine (25 mg, 0.24 mmol) were dissolved in DMF (1 mL) The reaction mixture was stirred at 100° C. for 2 h in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude product was purified by Pre-TLC (MeOH:DCM=1:20, V:V) to give white solid compound 19 (12 mg, yield 46%). MS m/z 546.1 [M+H]+.
The compound 4d (150 mg, 0.48 mmol) was dissolved in pyrdine (0.2 mL). Then 1d (58 mg, 0.24 mmol) was added. The reaction was stirred at 120° C. for 1 h in a sealed tube. The reaction was concentrated. The crude was purified by column chromatography to give white solid compound 19′a (10 mg, yield 8H). MS m/z 508.9 [M+H]+.
The compound 19′a (10 mg, 0.02 mmol), cesium carbonate (19 mg, 0.06 mmol) and di (triphenylphosphine) palladium dichloride (1.4 mg, 0.002 mmol) and 3-alkynylpyridine (10 mg, 0.10 mmol) were dissolved in DMF (1 mL) The reaction mixture was stirred at 100° C. for 2 h in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by Pre-TLC to give white solid compound 19′ (2 mg, yield 19%). 1H NMR (500 MHz, DMSO-d6) δ 14.01 (s, 1H), 8.77 (d, J=1.1 Hz, 1H), 8.60 (dd, J=4.8, 1.3 Hz, 1H), 8.02-7.97 (m, 1H), 7.86 (d, J=1.6 Hz, 1H), 7.51-7.44 (m, 2H), 6.69-6.52 (m, 4H), 6.30 (t, J=1.9 Hz, 1H), 5.41 (s, 2H), 3.83 (s, 3H), 3.78 (s, 3H). MS m/z 532.0 [M+H]+.
The compound 4c (30 mg, 0.06 mmol), cesium carbonate (60 mg, 0.18 mmol), di (triphenylphosphine) palladium dichloride (2.8 mg, 0.003 mmol) and 3-ethynyl-1-azacyclobutanoic acid tert butyl ester (41 mg, 0.23 mmol) were dissolved in DMF (1 mL) The reaction mixture was stirred at 100° C. for 2 h in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by Pre-TLC to give white solid compound 20 (25 mg, yield 70%). 1H NMR (500 MHz, DMSO-d6) δ 9.81 (s, 1H), 7.87 (d, J=1.6 Hz, 1H), 7.50 (d, J=1.0 Hz, 1H), 6.85-6.80 (m, 3H), 6.75 (s, 1H), 6.34-6.24 (m, 1H), 5.45 (s, 2H), 4.21-4.12 (m, 2H), 3.90-3.81 (m, 5H), 3.77 (s, 6H), 3.73-3.62 (m, 1H), 1.38 (s, 9H). MS m/z 624.1 [M+H]+.
The compound 20 (7 mg, 0.01 mmol) was dissolved in DCM (1 mL). Then trifluoroacetic acid (0.3 mL) was added. The reaction was stirred at RT for 2 h. The mixture was concentrated under reduced pressure to afford compound 21 (6.9 mg, yield 100%). 1H NMR (500 MHz, DMSO-d6) δ 9.92 (s, 1H), 8.86 (s, 2H), 7.88 (d, J=1.7 Hz, 1H), 7.50 (d, J=1.1 Hz, 1H), 6.86-6.72 (m, 4H), 6.30 (t, J=1.9 Hz, 1H), 5.45 (s, 2H), 4.31-4.15 (m, 2H), 4.11-3.92 (m, 3H), 3.86 (s, 3H), 3.77 (s, 6H). MS m/z 524.1 [M+H]+.
The compound 21 trifluoroacetate salt (20 mg, 0.03 mmol) was dissolved in DCM (1 mL). Then formaldehyde solution (0.1 mL, 37%) was added. The reaction mixture was stirred at RT for 1 h and Sodium triacetoxyborohydride (40 mg, 0.19 mmol) was added. The reaction was stirred at RT for another 1 h. The mixture was concentrated under reduced pressure. The crude was purified by Pre-TLC (MeOH:DCM=1:10) to give white solid compound 22 (3 mg, yield 17%). 1H NMR (500 MHz, DMSO-d6) δ 9.82 (s, 1H), 7.87 (s, 1H), 7.50 (s, 1H), 6.92-6.62 (m, 4H), 6.38-6.19 (m, 1H), 5.44 (s, 2H), 3.85 (s, 3H), 3.74 (s, 6H), 3.64-3.57 (m, 2H), 3.48-3.40 (m, 1H), 3.17-3.09 (m, 2H), 2.27 (s, 3H). MS m/z 538.0 [M+H]+.
The compound 18d (40 mg, 0.08 mmol), cesium carbonate (80 mg, 0.24 mmol) and di (triphenylphosphine) palladium dichloride (2.8 mg, 0.004 mmol) and 1-(Trimethylsilyl) propargyl (45 mg, 0.4 mmol) were dissolved in DMF (1 mL) The reaction mixture was stirred at 100° C. for 2 h in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by Pre-TLC (MeOH:DCM=1:20) to give white solid compound 23 (3 mg, yield 8%). 1H NMR (500 MHz, CDCl3) δ 8.18 (s, 1H), 7.64 (s, 1H), 7.50 (s, 1H), 7.35 (t, J=8.1 Hz, 1H), 7.16 (d, J=7.8 Hz, 1H), 6.91-6.75 (m, 2H), 6.59 (s, 1H), 6.38 (s, 1H), 5.45 (s, 2H), 3.98 (s, 3H), 3.87 (s, 3H), 2.13 (s, 3H). MS m/z 452.9 [M+H]+.
The compound 11b (25 mg, 0.05 mmol), cesium carbonate (49 mg, 0.15 mmol) and di (triphenylphosphine) palladium dichloride (2.0 mg, 0.003 mmol) and 3-ethynyl-1-azacyclobutanoic acid tert butyl ester 24a (28 mg, 0.15 mmol) were dissolved in DMF (1 mL) The reaction mixture was stirred at 100° C. for 2 h in a sealed tube under N2. The mixture was quenched with water and extracted with ethyl acetate (5 mL×3). The collected organic phase was washed with sat·NaCl, dried with anhydrous sodium sulfate, filterated and concentrated under reduced pressure. The crude was purified by Pre-TLC to give white solid compound 24b (20 mg, yield 67%). MS m/z 594.0 [M+H]+.
The compound 24b (20 mg, 0.03 mmol) was dissolved in DCM (1 mL). Then trifluoroacetic acid (0.3 mL) was added. The reaction was stirred at RT for 2 h. The mixture was concentrated under reduced pressure to afford compound 24c (20 mg, yield 100%). MS m/z 494.0 [M+H]+.
The compound 24c (20 mg, 0.03 mmol) was dissolved in DCM (1 mL). Then formaldehyde solution (0.1 mL, 37%) was added. The reaction was stirred at RT for 1 h and Sodium triacetoxyborohydride (40 mg, 0.19 mmol) was added. The reaction was stirred at RT for another 1 h. The mixture was concentrated under reduced pressure. The crude was purified by Pre-TLC (MeOH:DCM=1:10) to give white solid compound 24 (3 mg, yield 18%). 1H NMR (500 MHz, DMSO) δ 10.19 (br, 1H), 7.85 (d, J=2.2 Hz, 1H), 7.74 (d, J=8.0 Hz, 1H), 7.50-7.46 (m, 1H), 7.08-7.04 (m, 1H), 7.01 (dd, J=8.0, 1.3 Hz, 1H), 6.66 (s, 1H), 6.60 (s, 1H), 6.28 (t, J=2.0 Hz, 1H), 5.39 (s, 2H), 3.92 (t, J=7.6 Hz, 2H), 3.81 (s, 3H), 3.74 (s, 3H), 3.69-3.61 (m, 1H), 3.61-3.52 (m, 2H), 2.52 (s, 3H). MS m/z 508.0 [M+H]+.
1× experimental buffer (modified Tris buffer) was prepared, and the compounds were dissolved with 100% DMSO to form 10 mM reservoir solution and diluted according to a certain concentration gradient. The compounds were transferred to a 384-well plate using an automatic sampler Echo, and the final concentration of DMSO was 1%. 1× KAT6A enzyme (catalytic domain) solution was prepared. A mixed solution of [3H]-acetyl-CoA (PERKIN ELMER, Cat. No. NET290250UC) and the substrate peptide H3 (1-21) was prepared. 10 μL of the enzyme solution was transferred to the 384-well plate and incubated at room temperature for 15 minutes. 10 μL of the mixed solution of [3H]-acetyl-CoA and the substrate peptide H3 (1-21) were added to the plate to initiate the reaction. The plate was incubated at room temperature for 1 hour. 10 μL of the stop solution was added to terminate the reaction. 25 μL of the reaction solution was transferred to a Flashplate (Perkin Elmer, Cat. No. SMP410A001PK) and incubated at room temperature for 1 hour. The plate was read using Microbeta. The inhibition rate of the compounds was calculated using the formula of Excel: inhibition %=(Max-Signal)/(Max−Min)*100%. Where max is the value of the DMSO control, min is the value of the non-enzymatic control, and signal is the value of the well where the compound was tested. The curve was fitted using XLFit software and IC50 was calculated. The activities of some representative compounds are shown in Table 1.
Instruments: XEVO TQ-S LC/MS instrument was produced by Waters. All measurement data were collected and processed by Masslynx V4.1 software, and the data were calculated and processed by Microsoft Excel. The statistical moment method of WinNonLin 8.0 software was used to calculate the pharmacokinetic parameters. These mainly include kinetic parameters Tmax, T1/2, Cmax, AUC0-24 h etc. Column: ACQUITY UPLC BEH C18 (2.1 mm×50 mm, 1.7 μm); column temperature 40° C.; mobile phase A is water (0.1% formic acid), mobile phase B is acetonitrile, flow rate is 0.350 mL/min, gradient elution is adopted, the elution gradient is 0.50 min: 10% B; 1.50 min: 90% B; 2.50 min: 90% B; 2.51 min: 10% B; 3.50 min: stop. Injection volume: 1 μL.
Animals: 3 male SD rats with a weight range of 200-220 g were purchased and kept in the laboratory of the Experimental Animal Center for 2 days and then used. They were fasted for 12 hours predose and 4 hours after dosing. Drinking water was free during the test. After the rats were gavage, blood samples were taken according to the established time point.
Solvent: 0.4% ethanol+0.4% Tween80+99.2% (0.5% methylcellulose M450). Preparation of the solution for intragastrical administration: the compounds were accurately weighed and added to the solvent, and ultrasound was used at room temperature for 5 minutes to completely dissolve the drug, and a 0.3 mg/mL medicinal solution was prepared.
Pharmaceutical samples: Generally, multiple samples with similar structures (with a molecular weight difference of more than 2 units) are taken, accurately weighed, and administered together (cassette PK). In this way, multiple compounds can be screened at the same time and their oral absorption rates can be compared. A single administration was also used to study the pharmacokinetics of the drug sample in rats.
After intragastrical administration, blood was taken from the orbit at 0.25, 0.5, 1, 2, 4, 8, 10 and 24 hours. Then 50 μL of the plasma sample was taken and added 200 μL of acetonitrile (including internal standard control verapamil 2 ng/mL), vortexed for 3 min and centrifuged at 20000 rcf, 4° C. for 10 min. The supernatant plasma was used for LC-MS/MS analysis.
The compounds were accurately weighed to prepare different concentrations, and quantitative analysis on mass spectrometry was performed to establish a standard curve, and then the concentration of the above-mentioned compound in the plasma was tested to obtain its concentration at different time points. All measurement data were collected and processed by relevant software, and the statistical moment method was used to calculate the pharmacokinetic parameters (mainly including kinetic parameters Tmax, T1/2, Cmax, AUC0-24 h etc). The kinetic parameters of some representative compounds are shown in Table 2.
All references mentioned in the present invention are cited as references in this application, just as each reference is cited separately. In addition, it should be understood that after reading the above lecture content of the present invention, those skilled in the art can make various modifications or modifications to the present invention, and these equivalent forms also fall within the scope of the claims attached to the present application.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202110758732.7 | Jul 2021 | CN | national |
| 202210043389.2 | Jan 2022 | CN | national |
| 202210190640.8 | Feb 2022 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2022/104005 | 7/5/2022 | WO |
