A pyrimidine derivative, its preparation method and uses

Abstract

This application provides compounds of formula (I), their stereoisomers, or pharmaceutically acceptable salts thereof, and pharmaceutical compositions comprising them. The compounds, their stereoisomers, or pharmaceutically acceptable salts thereof are highly potent and specific CDK7 inhibitors capable of treating malignant tumor-related diseases.

Description

Technical Field

[0001] This application relates to the field of medicinal chemistry, specifically to a pyrimidine derivative or a pharmaceutically acceptable salt thereof as a cyclin-dependent kinase 7 (CDK7) inhibitor, its preparation method, and its use. Background Technology

[0002] Cyclin-dependent kinase 7 (CDK7) is a serine-threonine kinase that, in combination with cyclins, plays a crucial role in gene transcription and regulation. Mammalian cyclin-activated kinase (CAK), composed of CDK7, Cyclin H, and MAT1, can activate other CDKs... s Phosphorylation of the T-loops of CDK1, 2, 4, and 6 coordinates cell cycle progression. On the other hand, CDK7, as part of the multi-subunit universal transcription factor TFiIH complex, participates in the regulation of transcription. It phosphorylates the Ser residues at positions 5 and 7 of the C-terminal domain (CTD) of the RNA polymerase II subunit, thereby initiating transcription.

[0003] Studies have shown that CDK7 is closely related to the occurrence and development of various tumors. For example, it promotes high CDK7 expression through enhancers in tumor tissues such as triple-negative breast cancer, high-grade serous ovarian cancer, and small cell lung cancer. Therefore, CDK7 is considered a potential drug target for the treatment of malignant tumors, and the development of a highly effective and specific CDK7 inhibitor is of great significance for the treatment of malignant tumor-related diseases. Summary of the Invention

[0004] This application relates to a pyrimidine derivative as a CDK7 inhibitor, and more particularly to a pyrimidine derivative thereof, its preparation method and its pharmaceutical application, especially the use of the pyrimidine derivative shown in Formula I below in the preparation of drugs for CDK7-mediated diseases, and more specifically, in the preparation of drugs suitable for tumors.

[0005] One aspect of this application provides compounds with the structure shown in Formula I, their stereoisomers, or pharmaceutically acceptable salts thereof:

[0006]

[0007] in,

[0008] Ring A is selected from cyclic hydrocarbon groups or heterocyclic hydrocarbon groups, wherein the cyclic hydrocarbon group or heterocyclic hydrocarbon group can be selected from monocyclic, bicyclic, bridged, or spirocyclic rings;

[0009] X 1 Selected from CH or N;

[0010] R1 Selected from hydrogen, halogens, C1-C6 alkyl, C3-C8 cycloalkyl,

[0011] R 2 The group is selected from halogen, cyano, oxo, hydroxy, substituted or unsubstituted amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy is unsubstituted or substituted by one or more substituents, wherein the substituents are halogen, cyano, hydroxy, nitrogen, C1-C6 alkyl or C1-C6 alkoxy;

[0012] R 3 R 4 Each of the following groups is independently selected from hydrogen, C1-C6 alkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C3-C8 cycloalkyl, 3-8 membered heterocyclic alkyl, aryl or heteroaryl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, 3-8 membered heterocyclic alkyl, aryl or heteroaryl is unsubstituted or substituted by one or more substituents, wherein the substituents are halogen, cyano, hydroxyl, nitrogen, C1-C6 alkyl or C1-C6 alkoxy;

[0013] m is selected from 0, 1, 2, 3, 4 or 5.

[0014] Preferably, the compound with the structure shown in Formula I, its stereoisomers, or its pharmaceutically acceptable salts,

[0015] in,

[0016] Ring A is selected from 4-8 membered cyclic hydrocarbon groups or 4-10 membered heterocyclic hydrocarbon groups, wherein the cyclic hydrocarbon group or heterocyclic hydrocarbon group can be selected from monocyclic, bicyclic, bridged, or spirocyclic rings, and the heterocyclic hydrocarbon group contains at least one nitrogen atom as a heteroatom;

[0017] X 1 Selected from CH or N;

[0018] R 1 Selected from hydrogen, halogens, C1-C6 alkyl, C3-C8 cycloalkyl,

[0019] R 2The group is selected from halogen, cyano, oxo, hydroxy, substituted or unsubstituted amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy is unsubstituted or substituted by one or more substituents, wherein the substituents are halogen, cyano, hydroxy, amino, C1-C6 alkyl or C1-C6 alkoxy;

[0020] R 3 R 4 Each of the following is independently selected from hydrogen, C1-C6 alkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C3-C8 cycloalkyl, 3-8 heterocyclic alkyl, 6-10 aryl or 5-10 heteroaryl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, 3-8 heterocyclic alkyl, 6-10 aryl or 5-10 heteroaryl is unsubstituted or substituted by one or more substituents, wherein the substituents are halogen, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy;

[0021] m is selected from 0, 1, 2, 3, 4 or 5.

[0022] In some embodiments, ring A is selected from 5-7 membered cycloalkyl or 5-7 membered heterocycloalkyl, wherein the heterocycloalkyl contains 1-2 heteroatoms selected from N or O.

[0023] In some embodiments, ring A is cyclopentyl, cyclohexyl, cycloheptyl, tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl, piperidinyl, oxetyl-heptyl, or aziryl-heptyl.

[0024] In some implementations, X 1 It can be CH or N, with CH being preferred.

[0025] In some implementations, R 1 Selected from hydrogen, halogens (e.g., fluorine, chlorine, bromine, iodine), C1-C5 alkyl groups (e.g., C1-C4 alkyl groups, C1-C3 alkyl groups), and C3-C6 cycloalkyl groups (e.g., C3-C5 cycloalkyl groups, C3-C4 cycloalkyl groups). and

[0026] R 3 R 4Each of the following is independently selected from hydrogen, C1-C5 alkyl (e.g., C1-C4 alkyl, C1-C3 alkyl), C1-C5 alkyl carbonyl (e.g., C1-C4 alkyl carbonyl, C1-C3 alkyl carbonyl), C1-C5 alkoxy carbonyl (e.g., C1-C4 alkoxy carbonyl, C1-C3 alkoxy carbonyl), 4-7 membered heterocyclic alkyl, 6 membered aryl, or 5-7 membered heteroaryl, wherein the 4-7 membered heterocyclic alkyl, 6 membered aryl, or 5-7 membered heteroaryl is unsubstituted or substituted by one or more substituents, wherein the substituents are hydroxyl, C1-C5 alkyl (e.g., C1-C4 alkyl, C1-C3 alkyl), or C1-C5 alkoxy (e.g., C1-C4 alkoxy, C1-C3 alkoxy), and wherein the heterocyclic alkyl and heteroaryl contain 1-3 (e.g., 1-2) heteroatoms selected from N, O, or S.

[0027] In some implementations, R 1 Selected from hydrogen, fluorine, chlorine, bromine, C1-C4 alkyl (e.g., methyl, ethyl, propyl, or butyl), C3-C5 cycloalkyl (e.g., cyclopropyl, cyclobutyl, or cyclopentyl), or and

[0028] R 3 R 4 Each of the following is independently selected from hydrogen, C1-C5 alkyl carbonyl (e.g., C1-C4 alkyl carbonyl, C1-C3 alkyl carbonyl), C1-C5 alkoxy carbonyl (e.g., C1-C4 alkoxy carbonyl, C1-C3 alkoxy carbonyl), or 5-7-membered heteroaryl, wherein the 5-7-membered heteroaryl is unsubstituted or substituted by one or more substituents, wherein the substituents are C1-C5 alkyl (e.g., C1-C4 alkyl, C1-C3 alkyl), and the heteroaryl contains 1-2 heteroatoms selected from N, O, or S; preferably, R 3 R 4 Each of the following is independently selected from hydrogen, C1-C4 alkoxycarbonyl (e.g., C1-C3 alkoxycarbonyl), or 5-6 heteroaryl, wherein the 5-6 heteroaryl is unsubstituted or substituted by one or more substituents, wherein the substituents are C1-C4 alkyl (e.g., C1-C3 alkyl), and wherein the heteroaryl contains 1-2 heteroatoms selected from N, O, or S.

[0029] In some implementations, R 2 Selected from halogen, cyano, oxo, hydroxy, amino, or C1-C5 alkyl (e.g., C1-C4 alkyl, C1-C3 alkyl), and m is 0, 1, 2, or 3; preferably, R 2 It is selected from halogen, hydroxyl, amino or C1-C3 alkyl, and m is 0, 1 or 2 (e.g. m is 0).

[0030] Preferably, this application provides compounds with the structure shown in Formula II, their stereoisomers, or pharmaceutically acceptable salts thereof.

[0031]

[0032] in,

[0033] R 1 Selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, cyclopropyl,

[0034] R 2 Selected from fluorine, chlorine, cyano, oxo, hydroxyl, methyl, ethyl, or isopropyl;

[0035] R 3 R 4 Each of the following groups is independently selected from hydrogen, methyl, ethyl, isopropyl, cyclopropyl, n-propyl, cyclobutyl, cyclopentyl, cyclohexyl, Phenyl or 5-6 heteroaryl, wherein the 5-6 heteroaryl may be further substituted by one or more C1-C3 alkyl groups;

[0036] m is selected from 0, 1, or 2.

[0037] In some embodiments, in compounds with the structure shown in Formula II, R 1 R 2 R 3 R 4 The definitions of and m are the same as those in the various embodiments of the compound with the structure shown in Formula I described above.

[0038] In some implementations, R 1 Selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, or (e.g., hydrogen, fluorine, chlorine, bromine, methyl, ethyl, cyclopropyl or) ); R 2 Selected from methyl, ethyl, or isopropyl (e.g., methyl); R 3 R 4 Selected independently from hydrogen, Alternatively, a 5-6 membered heteroaryl group may be further substituted with one or more (e.g., 1-3, 1-2) C1-C5 alkyl groups, wherein the heteroaryl group comprises 1-2 heteroatoms selected from N, O, or S (e.g., R). 3 R 4 Selected independently from hydrogen, Or a 5-membered heteroaryl group, wherein the 5-membered heteroaryl group may be further substituted by 1-2 C1-C3 alkyl groups, wherein the heteroaryl group contains 1-2 heteroatoms selected from N or O; m is selected from 0, 1 or 2 (e.g., 0).

[0039] More preferably, this application provides compounds with the structure shown in Formula III, their stereoisomers, or pharmaceutically acceptable salts thereof.

[0040]

[0041] in,

[0042] R 2 Selected from fluorine, chlorine, cyano, oxo, hydroxyl, methyl, ethyl, or isopropyl;

[0043] R 3 R 4 Each of the following groups is independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, Phenyl or 5-6 heteroaryl, wherein the 5-6 heteroaryl may be further substituted by one or more C1-C3 alkyl groups;

[0044] m is selected from 0, 1, or 2.

[0045] In some embodiments, in compounds with the structure shown in Formula III, R 1 R 2 R 3 R 4 The definitions of and m are the same as those in the various embodiments of the compound with the structure shown in Formula I described above.

[0046] In some implementations, R 2 Selected from methyl, ethyl, or isopropyl (e.g., methyl); R 3 R 4 Selected independently from hydrogen, Alternatively, a 5-6 membered heteroaryl group may be further substituted with one or more (e.g., 1-3, 1-2) C1-C5 alkyl groups, wherein the heteroaryl group comprises 1-2 heteroatoms selected from N, O, or S (e.g., R). 3 R 4 Selected independently from hydrogen, Or a 5-membered heteroaryl group, wherein the 5-membered heteroaryl group may be further substituted by 1-2 C1-C3 alkyl groups, wherein the heteroaryl group contains 1-2 heteroatoms selected from N or O; m is selected from 0, 1 or 2 (e.g., 0).

[0047] In some embodiments, this application provides compounds with the structure shown in Formula I, their stereoisomers, or pharmaceutically acceptable salts thereof, wherein

[0048] Ring A is selected from 5-7 membered cycloalkyl or 5-7 membered heterocycloalkyl, wherein the heterocycloalkyl contains 1-2 heteroatoms selected from N or O; preferably, ring A is cyclohexyl, pyrrolidinyl, piperidinyl or azirheptanyl (preferably piperidinyl);

[0049] X 1 It can be CH or N, with CH being preferred;

[0050] R 1 Selected from hydrogen, fluorine, chlorine, bromine, C1-C4 alkyl (e.g., methyl, ethyl, propyl, or butyl), C3-C5 cycloalkyl (e.g., cyclopropyl, cyclobutyl, or cyclopentyl), or

[0051] R 3 R 4 Each of the following is independently selected from hydrogen, C1-C5 alkyl carbonyl (e.g., C1-C4 alkyl carbonyl, C1-C3 alkyl carbonyl), C1-C5 alkoxy carbonyl (e.g., C1-C4 alkoxy carbonyl, C1-C3 alkoxy carbonyl), or 5-7 heteroaryl, wherein the 5-7 heteroaryl is unsubstituted or substituted by one or more (e.g., 1-3, 1-2) substituents, wherein the substituents are C1-C5 alkyl (e.g., C1-C4 alkyl, C1-C3 alkyl), and wherein the heteroaryl contains 1-2 heteroatoms selected from N, O or S;

[0052] R 2 It is selected from halogen, hydroxyl, amino or C1-C3 alkyl, and m is 0, 1 or 2 (e.g. m is 0).

[0053] In some embodiments, this application provides compounds with the structure shown in Formula I, their stereoisomers, or pharmaceutically acceptable salts thereof, wherein

[0054] Ring A is selected from 5-7 membered heterocyclic alkyl groups, wherein the heterocyclic alkyl group contains 1-2 heteroatoms selected from N or O; preferably, ring A is pyrrolidinyl, piperidinyl or azirheptanyl (preferably piperidinyl);

[0055] X 1 It can be CH or N, with CH being preferred;

[0056] R 1 Selected from hydrogen, fluorine, chlorine, bromine, C1-C4 alkyl (e.g., methyl, ethyl, propyl, or butyl), C3-C5 cycloalkyl (e.g., cyclopropyl, cyclobutyl, or cyclopentyl), or

[0057] R 3 R 4Each of the following is independently selected from hydrogen, C1-C5 alkoxycarbonyl (e.g., C1-C4 alkoxycarbonyl, C1-C3 alkoxycarbonyl), or 5-7 heteroaryl, wherein the 5-7 heteroaryl is unsubstituted or substituted by one or more (e.g., 1-3, 1-2) substituents, wherein the substituents are C1-C5 alkyl (e.g., C1-C4 alkyl, C1-C3 alkyl), and wherein the heteroaryl contains 1-2 heteroatoms selected from N, O or S;

[0058] m is 0.

[0059] In some embodiments, this application provides compounds with the structure shown in Formula I, their stereoisomers, or pharmaceutically acceptable salts thereof, wherein

[0060] Ring A is selected from 5-7 membered heterocyclic alkyl groups, wherein the heterocyclic alkyl group contains 1-2 heteroatoms selected from N or O; preferably, ring A is pyrrolidinyl, piperidinyl or azirheptanyl (preferably piperidinyl);

[0061] X 1 It can be CH or N, with CH being preferred;

[0062] R 1 Selected from hydrogen, fluorine, chlorine, bromine, C1-C4 alkyl (e.g., methyl, ethyl, propyl, or butyl), or

[0063] R 3 R 4 Each of the following is independently selected from hydrogen, C1-C5 alkoxycarbonyl (e.g., C1-C4 alkoxycarbonyl, C1-C3 alkoxycarbonyl), or 5-7 heteroaryl, wherein the 5-7 heteroaryl is unsubstituted or substituted by one or more (e.g., 1-3, 1-2) substituents, wherein the substituents are C1-C5 alkyl (e.g., C1-C4 alkyl, C1-C3 alkyl), and wherein the heteroaryl contains 1-2 heteroatoms selected from N, O or S;

[0064] m is 0.

[0065] More preferably, this application provides compounds with the structure shown in Formula I, their stereoisomers, or pharmaceutically acceptable salts thereof, said compounds being selected from:

[0066]

[0067]

[0068] This application also covers solutions obtained by any combination, deletion or substitution of the above-described embodiments and preferred solutions.

[0069] Another aspect of this application provides a method for preparing compounds with the structure shown in Formula I above, comprising:

[0070]

[0071] Where LG represents the leaving group, PG represents the protecting group, and R 1 R 2 X 1 The ring A and m are as defined above in the compound with the structure shown in Formula I; preferably, the leaving group is selected from a halogen atom, methanesulfonyloxy group or p-toluenesulfonyloxy group; the protecting group is selected from 2-(trimethylsilyl)ethoxymethyl or tert-butoxycarbonyl.

[0072] (1) Compound I-1 and compound I-2 were subjected to a substitution reaction to obtain compound I-3.

[0073] The reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from tetrahydrofuran, 1,4-dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, acetonitrile, and any combination thereof, with N,N-dimethylformamide being preferred.

[0074] The reaction is preferably carried out in the presence of a suitable base. The base may be selected from triethylamine, pyridine, 4-dimethylaminopyridine, diisopropylethylamine, potassium carbonate, cesium carbonate, sodium carbonate, and preferably diisopropylethylamine.

[0075] The reaction is preferably carried out at a suitable temperature, preferably 100-150°C. The reaction is preferably carried out for a suitable time, for example, 8-12 hours.

[0076] (2) Compound I-3 and compound I-4 were coupled together to obtain compound I-5.

[0077] The reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from tetrahydrofuran, 1,4-dioxane, toluene, acetonitrile, ethanol, water, and any combination thereof, preferably a combination of 1,4-dioxane and water.

[0078] The reaction is preferably carried out in the presence of a suitable catalyst. The catalyst may be selected from Pd(dppf)Cl2, Pd(OAc)2, Pd2(dba)3, Pd(PPh3)4, with Pd(dppf)Cl2 being preferred.

[0079] The reaction is preferably carried out in the presence of a suitable base. The base may be selected from triethylamine, pyridine, 4-dimethylaminopyridine, diisopropylethylamine, potassium carbonate, cesium carbonate, sodium carbonate, with potassium carbonate being preferred.

[0080] The reaction is preferably carried out at a suitable temperature, preferably 80-120°C. The reaction is preferably carried out for a suitable time, for example, 8-12 hours.

[0081] (3) Compound I-5 was subjected to a deprotection reaction to obtain the compound shown in Formula I.

[0082] The reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from tetrahydrofuran, 1,4-dioxane, dichloromethane, ethyl acetate, and preferably dichloromethane.

[0083] The reaction is preferably carried out in the presence of a suitable acid. The acid may be hydrochloric acid or trifluoroacetic acid, with trifluoroacetic acid being preferred.

[0084] The reaction is preferably carried out at a suitable temperature, preferably 20-50°C. The reaction is preferably carried out for a suitable time, for example, 4-6 hours.

[0085] The specific conditions for each of the above reaction steps are well known in the art and are not specifically limited thereto in this application. If the teachings of this application are combined with common knowledge in the art, those skilled in the art can selectively replace the substituents in the general formula to prepare different compounds, and such selections and substitutions are all within the scope of protection of this application.

[0086] This application also relates to a pharmaceutical composition comprising the above-described compound of formula I, its stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0087] This application also relates to a pharmaceutical composition comprising the above-described compound of formula II, its stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0088] This application also relates to a pharmaceutical composition comprising the above-described compound of formula III, its stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0089] This application also relates to the use of the above-described Formula I compound, its stereoisomers or pharmaceutically acceptable salts thereof, or the above-described pharmaceutical compositions in the preparation of a medicament for the prevention or treatment of CDK7-mediated diseases or conditions. Alternatively, this application also relates to a Formula I compound, its stereoisomers or pharmaceutically acceptable salts thereof, or the above-described pharmaceutical compositions for the prevention or treatment of CDK7-mediated diseases or conditions. Alternatively, this application also relates to a method for the prevention or treatment of CDK7-mediated diseases or conditions, comprising administering to a subject in need a Formula I compound, its stereoisomers or pharmaceutically acceptable salts thereof, or the above-described pharmaceutical compositions. In this document, the subject may be a mammal, such as a human, monkey, cat, dog, pig, sheep, cow, horse, rabbit, mouse, etc.

[0090] In this document, CDK7-mediated diseases or conditions refer to those diseases or conditions for which desired clinical benefits can be obtained by inhibiting CDK7. In some embodiments, the CDK7-mediated diseases or conditions are selected from tumors or cancers, such as breast cancer, pancreatic cancer, ovarian cancer, colorectal cancer, lung cancer, prostate cancer, lymphoma, malignant sarcoma, cervical cancer, oral cancer, brain cancer, gastric cancer, liver cancer, skin cancer, bone cancer, kidney cancer, bladder cancer, fallopian tube tumors, peritoneal tumors, melanoma, glioma, glioblastoma, papillary malignancy, head and neck tumors, myeloma, or leukemia.

[0091] This application also relates to the use of the above-described Formula I compound, its stereoisomers or pharmaceutically acceptable salts thereof, or the above-described pharmaceutical composition in the preparation of a CDK7 inhibitor. Alternatively, this application also relates to the use of the above-described Formula I compound, its stereoisomers or pharmaceutically acceptable salts thereof, or the above-described pharmaceutical composition as a CDK7 inhibitor. Alternatively, this application also relates to a method of inhibiting CDK7, comprising administering to a subject in need the above-described Formula I compound, its stereoisomers or pharmaceutically acceptable salts thereof, or the above-described pharmaceutical composition.

[0092] This application discovers a novel class of CDK7 inhibitors with the structure shown in Formula I, which exhibit good CDK7 inhibitory activity and can demonstrate the desired antitumor effect. Detailed Implementation

[0093] To make the aspects and technical solutions of this application clearer, the following describes the application in further detail with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the application. Furthermore, any specific experimental methods not mentioned in the following embodiments are performed according to conventional experimental methods.

[0094] Definitions and General Descriptions

[0095] Unless otherwise stated, the terms used in this application have the following meanings. A particular term should not be considered uncertain or unclear unless specifically defined, but should be understood in accordance with its ordinary meaning in the art.

[0096] When the covalent bonds in certain structural units or groups in this application are not connected to specific atoms, it means that the covalent bonds can be connected to any atom in the structural unit or group, as long as the rules of valence bond connection are not violated.

[0097] In this document, unless otherwise stated, "hydrocarbon group" refers to a group consisting of only two types of atoms: carbon and hydrogen, including saturated hydrocarbon groups (also known as alkyl groups) and unsaturated hydrocarbon groups such as alkenes and alkynes.

[0098] In this document, unless otherwise stated, "cyclic hydrocarbon group" refers to a cyclic group consisting of only carbon and hydrogen atoms, including saturated cyclic hydrocarbon groups (also known as cycloalkyl groups) and unsaturated cyclic hydrocarbon groups, such as cyclic hydrocarbon groups containing carbon-carbon double bonds.

[0099] In this article, unless otherwise stated, "heterocyclic hydrocarbon group" refers to a group that contains heterocyclic atoms such as nitrogen, oxygen, and sulfur atoms in addition to carbon and hydrogen atoms to participate in the ring formation, including saturated heterocyclic hydrocarbon groups (also known as heterocyclic alkyl groups) and unsaturated heterocyclic hydrocarbon groups, such as heterocyclic hydrocarbon groups containing carbon-carbon double bonds.

[0100] In this document, unless otherwise stated, "alkyl" refers to a saturated hydrocarbon group consisting only of carbon and hydrogen atoms, linked by single bonds between carbons and between carbons and hydrogens. Alkyl groups can be straight-chain or branched. Representative branched alkyl groups have one, two, or three branches. Alkyl groups may optionally be substituted by one or more substituents as defined herein. Alkyl groups include, but are not limited to, methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl, and tert-butyl), pentyl (n-pentyl, isopentyl, and neopentyl), and hexyl. Alkyl groups may also be part of other groups, such as (C1-C6 alkyl)-O-, etc.

[0101] In this document, unless otherwise stated, "cycloalkyl" refers to a fully saturated carbon ring that may exist as a monocyclic, bridged, or spirocyclic ring. Unless otherwise indicated, cycloalkyl in this document may be C3-C8 cycloalkyl, such as 3-membered, 4-membered, 5-membered, 6-membered, 7-membered, or 8-membered rings. Non-limiting examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0102] In this document, unless otherwise stated, "heterocyclic alkyl" refers to a saturated or non-aromatic partially saturated ring containing at least one heteroatom and having a single or multiple rings (fused, bridged, spirofused); where the heteroatom refers to nitrogen, oxygen, sulfur, etc. It typically represents a monovalent saturated or partially unsaturated monocyclic or polycyclic ring system containing 1, 2, or 3 cyclic heteroatoms selected from N, O, and S, with the remaining cyclic atoms being carbon.

[0103] In this document, unless otherwise stated, "aromatic ring" refers to an aromatic group having multiple carbon atoms. Aryl groups are typically monocyclic, bicyclic, or tricyclic aryl groups having multiple carbon atoms. Furthermore, as used herein, the term "aryl" refers to an aromatic substituent that can be a single aromatic ring or multiple aromatic rings fused together.

[0104] In this document, unless otherwise stated, "aromatic heterocycle" is an aromatic unsaturated ring containing at least one heteroatom, where the heteroatom refers to a nitrogen atom, oxygen atom, or sulfur atom, etc.; generally, it refers to an aromatic monocyclic or bicyclic hydrocarbon containing multiple ring atoms, one or more of which are substituted by heteroatoms selected from O, N, and S. Preferably, it contains one to three heteroatoms. Representative heterocyclic aryl groups include, for example: pyridyl, indolyl, quinoxalinyl, quinolinyl, isoquinolinyl, benzothiophene, benzofuranyl, benzothiophene, benzopyranyl, benzothiapyranyl, furanyl, pyrroleyl, thiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazole, thiophene, oxadiazolyl, benzimidazole, benzothiazolyl, benzoxazolyl, etc.

[0105] In this article, unless otherwise stated, "unsaturated" means that the group or molecule contains carbon-carbon double bonds, carbon-nitrogen double bonds, etc.

[0106] In this document, unless otherwise stated, "alkoxy" refers to -O-alkyl.

[0107] In this article, unless otherwise stated, "halogen" means fluorine, chlorine, bromine or iodine.

[0108] In this document, unless otherwise stated, “cycloalkyloxy” refers to cycloalkyl-O-.

[0109] In this document, unless otherwise stated, “substituted or unsubstituted amino” encompasses an unsubstituted amino group or an amino group substituted with a group selected from the group consisting of C1-C6 alkyl, halo-C1-C6 alkyl, C3-C8 cycloalkyl, or halo-C3-C8 cycloalkyl.

[0110] In this document, unless otherwise stated, the term "C" is used. m -C n "" means that the part modified by the term has mn carbon atoms (n is greater than m, and both are integers). For example, C1-C6 means that the part it modifies has 1-6 carbon atoms, such as 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms.

[0111] In this document, unless otherwise stated, "mn-membered (hetero)cyclic hydrocarbon group" refers to the total number of carbon atoms and heteroatoms involved in cyclization in the modified portion, where the heteroatoms can be nitrogen atoms, sulfur atoms, oxygen atoms, etc. (n is greater than m, and both are integers). For example, 4-8-membered cyclic alkenes indicate that the modified cyclic structure has 4, 5, 6, 7, or 8 carbon atoms; 4-10-membered heterocyclic hydrocarbon groups indicate that the total number of carbon atoms and heteroatoms in the modified cyclic structure is 4, 5, 6, 7, 8, 9, or 10.

[0112] In this application, the terms “comprising,” “including,” and “containing,” and their equivalents, are to be understood in an open, non-exclusive sense, meaning “including but not limited to,” implying that in addition to the listed elements, components, and steps, other unspecified elements, components, and steps may also be included. In this document, unless the context clearly indicates otherwise, singular terms encompass plural referents, and vice versa. Similarly, unless the context clearly indicates otherwise, the word “or” is intended to include “and.”

[0113] Unless otherwise stated, the parameter values ​​representing the amount of components, physicochemical properties, or reaction conditions, etc., shall be understood to be modified by the term "about" in all cases. When the term "about" is used to describe this application, the term "about" indicates an existing error value, such as a variation within ±5%, for example ±1%, or ±0.1% of a particular value.

[0114] In this application, when the chemical name and structural formula are inconsistent, the structural formula shall prevail, unless the chemical name rather than the structural formula can be inferred to be correct from the context.

[0115] The abbreviations used in this article have the following meanings:

[0116] abbreviation meaning <![CDATA[DMSO-d6]]> Hexadecimalized dimethyl sulfoxide TMS Tetramethylsilane <![CDATA[ 1 H NMR]]> Hydrogen spectrum MS mass spectrometry s Single peak d Double peak t Triple Peak q Four Peaks dd Double peak m Multiplets br Broad Peak J Coupling constant Hz hertz

[0117] The structure of a compound can be determined by mass spectrometry (MS) or nuclear magnetic resonance (NMR). 1 It can be determined by H NMR.

[0118] 1H NMR spectrum ( 1 H NMR shift (δ) is given in parts per million (ppm); nuclear magnetic resonance (NMR) 1 The ¹H NMR (hydrocarbon) measurements were performed using a Bruker Avance-400 NMR spectrometer. The solvent was deuterated dimethyl sulfoxide (DMSO-d6), and the internal standard was tetramethylsilane (TMS). Chemical shifts were expressed as 10⁻¹⁰ NMR values. -6 (ppm) is given as the unit.

[0119] Mass spectrometry (MS) measurements were performed using a FINNIGAN LCQAd (ESI) mass spectrometer (manufacturer: Therm, model: Finnigan LCQ advantage MAX).

[0120] Thin-layer silicone uses Yantai Huanghai HSGF254 or Qingdao GF254 silicone sheets.

[0121] Column chromatography typically uses Yantai Huanghai silica gel with a mesh size of 200-300 as the carrier.

[0122] Unless otherwise specified in this application, all reactions mentioned herein are carried out under a nitrogen atmosphere.

[0123] The term "nitrogen atmosphere" in this application refers to, for example, connecting a reaction flask to a nitrogen balloon with a volume of 1 L.

[0124] The term "hydrogen atmosphere" in this application refers to, for example, connecting a reaction vessel to a 1L hydrogen balloon.

[0125] Unless otherwise specified in this application, the solutions mentioned in the reactions described herein are aqueous solutions.

[0126] The term "room temperature" in this application refers to a temperature between 10°C and 25°C.

[0127] For purposes of description and disclosure, all patents, patent applications, and other identified publications are expressly incorporated herein by reference. These publications are provided solely because their publication predates the filing date of this application. All statements regarding the dates of these documents or representations of their contents are based on information available to the applicant and do not constitute any acknowledgment of the accuracy of the dates or contents of these documents. Furthermore, in any country, any reference to these publications herein does not constitute an endorsement that such publication is part of the general knowledge in the art.

[0128] The present application will be described in detail below through embodiments; however, those skilled in the art will understand that the scope of protection of the present application is not limited thereto. Those skilled in the art can make various modifications, changes, combinations, etc., to the implementation methods and embodiments of the present application without departing from the spirit or scope of the present application, and the resulting adjusted solutions also fall within the scope of protection of the present application.

[0129] Example 1 Preparation of (S)-3-(2-(piperidin-3-amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridine-6-amine (1)

[0130]

[0131] Step 1: Preparation of (S)-3-((4-chloro-5-(trifluoromethyl)pyrimidin-2-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (1b)

[0132] Compound 1a (800 mg, 3.7 mmol) and (S)-3-aminopiperidine-1-carboxylic acid tert-butyl ester (890 mg, 4.4 mmol) were dissolved in N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (955 mg, 7.4 mmol) was added. The mixture was reacted at 80 °C for 4 hours. The reaction solution was diluted with water, extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to give 900 mg of the title compound, yield: 64.0%.

[0133] LC-MS(ESI)m / z(M+H) + 381.3

[0134] Step 2: Preparation of (S)-3-(2-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-6-chloro-1H-pyrrolo[2,3-b]pyridine-1-carboxylic acid tert-butyl ester (1c)

[0135] Compound 1b (900 mg, 2.36 mmol), 6-chloro-1-(tert-butoxycarbonyl)-1H-pyrrolo[2,3-b]pyridine-3-boronic acid pinacol ester (1.07 g, 2.84 mmol), Pd(dppf)Cl2 (174.16 mg, 0.24 mmol), and potassium carbonate (652.35 mg, 4.72 mmol) were dissolved in 1,4-dioxane (20 mL). Under nitrogen protection, the reaction mixture was heated to 100 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 541 mg of the title compound, yield: 38.4%.

[0136] LC-MS(ESI)m / z(M+H) + 597.1

[0137] Step 3: Preparation of (S)-3-(2-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-6-amino-1H-pyrrolo[2,3-b]pyridine-1-carboxylic acid tert-butyl ester (1d)

[0138] Compound 1c (100 mg, 0.17 mmol), carbamate (15 mg, 0.20 mmol), RuPhos-Pd-G3 (16.75 mg, 0.02 mmol), and cesium carbonate (110.77 mg, 0.34 mmol) were dissolved in toluene (5 mL). Under nitrogen protection, the reaction mixture was heated to 110 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 27.89 mg of the title compound, yield: 28.4%.

[0139] LC-MS(ESI)m / z(M+H) + 578.2

[0140] Step 4: Preparation of (S)-3-(2-(piperidin-3-amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridine-6-amine (1)

[0141] Compound 1d (25 mg, 0.04 mmol) was dissolved in dichloromethane (5 mL), and then trifluoroacetic acid (2 mL) was added. The reaction system was stirred at room temperature for 2 hours. The reaction system was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 7 mg of the title compound, yield: 46.7%.

[0142] LC-MS(ESI)m / z(M+H) + 378.2

[0143] 1 H NMR (400MHz, DMSO-d6) δ11.54(d, J=13.9Hz, 1H), 8.54-8.45(m, 1.5H), 8.25(d , J=8.6Hz, 0.5H), 7.68-7.64 (m, 1H), 7.48 (s, 1H), 6.39 (d, J=8.6Hz, 1H), 5.81 (d, J=10.9Hz, 2H), 3.89 (s, 1H), 3.07 (d, J=11.2Hz, 1H), 2.82 (d, J=12.1Hz, 1H ), 2.48-2.40(m, 2H), 2.00-1.92(m, 1H), 1.68-1.64(m, 1H), 1.50-1.40(m, 2H).

[0144] Example 2 Preparation of (S)-3-(2-(piperidin-3-amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)aminocarbamate (2)

[0145]

[0146] Step 1: Preparation of (S)-3-(2-((1-(tert-butyloxycarbonyl)piperidin-3-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-6-methoxycarbonylamino-1H-pyrrolo[2,3-b]pyridine-1-carboxylic acid tert-butyl ester (2a)

[0147] Compound 1c (100 mg, 0.17 mmol), carbamate (15 mg, 0.20 mmol), RuPhos-Pd-G3 (16.75 mg, 0.02 mmol), and cesium carbonate (110.77 mg, 0.34 mmol) were dissolved in toluene (5 mL). Under nitrogen protection, the reaction system was heated to 110 °C and stirred for 4 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 48 mg of the title compound, yield: 44.4%.

[0148] LC-MS(ESI)m / z(M+H) + : 636.4

[0149] Step 2: Preparation of (S)-3-(2-(piperidin-3-amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)aminocarbamate (2)

[0150] Compound 2a (48 mg, 0.08 mmol) was dissolved in dichloromethane (5 mL), and then trifluoroacetic acid (2 mL) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 15 mg of the title compound, yield: 45.6%.

[0151] LC-MS(ESI)m / z(M+H) + : 436.2

[0152] 1 H NMR (400MHz, DMSO-d6) δ12.13 (s, 1H), 10.08 (d, J = 10.1Hz, 1H), 8.84 (d, J = 8.7Hz, 0.5H), 8.59-8.51 (m, 1.5H), 7.82-7.71 (m, 3H), 3.91 (s , 1H), 3.69 (s, 3H), 3.10 (d, J=11.9Hz, 1H), 2.85 (d, J=12.2Hz, 1H), 2.52-2.43 (m, 2H), 2.03-1.96 (m, 1H), 1.68 (s, 1H), 1.53-1.43 (m, 2H).

[0153] Example 3 Preparation of (S)-3,5-dimethyl-N-(3-(2-(piperidin-3-amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole-4-amine (3)

[0154]

[0155] Step 1: Preparation of (S)-3-(2-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-6-((3,5-dimethylisoxazol-4-yl)amino)-1H-pyrrolo[2,3-b]pyridine-1-carboxylic acid tert-butyl ester (3a)

[0156] Compound 1c (100 mg, 0.17 mmol), 3,5-dimethylisoxazol-4-amine (22.4 mg, 0.20 mmol), RuPhos-Pd-G3 (16.75 mg, 0.02 mmol), and cesium carbonate (110.77 mg, 0.34 mmol) were dissolved in toluene (5 mL). Under nitrogen protection, the reaction mixture was heated to 110 °C and stirred for 12 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 62.7 mg of the title compound, yield: 54.8%.

[0157] LC-MS(ESI)m / z(M+H) + 673.1

[0158] Step 2: Preparation of (S)-3,5-dimethyl-N-(3-(2-(piperidin-3-amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridin-6-yl)isoxazole-4-amine (3)

[0159] Compound 3a (60 mg, 0.09 mmol) was dissolved in dichloromethane (5 mL), and then trifluoroacetic acid (2 mL) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 32 mg of the title compound, yield: 76.2%.

[0160] LC-MS(ESI)m / z(M+H) + : 473.2

[0161] 1H NMR (400MHz, DMSO-d6) 611.87 (s, 1H), 8.60-8.52 (m, 1.5H), 8.32 (d, J=8.7Hz, 0.5H), 8.01 (d, J=5.0Hz, 1H), 7.79 (t, J=8.6Hz, 1H), 7.53 (d, J=6.8Hz, 1H), 6.5 8-6.55(m, 1H), 4.03(s, 1H), 3.24-3.18(m, 1H), 2.97(s, 1H), 2.68-2.58(m, 2H) , 2.28(s, 3H), 2.09(s, 3H), 2.03-1.97(m, 1H), 1.76(s, 1H), 1.55-1.50(m, 2H).

[0162] Example 4 Preparation of (S)-4-(6-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)-N-(piperidin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine (4)

[0163]

[0164] Compound 1c (50 mg, 0.08 mmol) was dissolved in dichloromethane (5 mL), and then trifluoroacetic acid (2 mL) was added. The reaction system was stirred at room temperature for 2 hours. The reaction system was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 22 mg of the title compound, yield: 66.2%.

[0165] LC-MS(ESI)m / z(M+H) + 397.1

[0166] 1 H NMR (400MHz, DMSO-d6) δ8.85 (d, J=8.3Hz, 0.5H), 8.63-8.55 (m, 1.5H), 7.95 (s, 1H), 7.87 (t, J=8.6Hz, 1H), 7.32-7.26 (m, 1H), 3.92-3. 86 (m, 1H), 3.13-3.04 (m, 1H), 2.83 (d, J=12.0Hz, 1H), 2.47-2.42 (m, 2H), 1.99-1.90 (m, 1H), 1.67 (d, J=8.5Hz, 1H), 1.50-1.42 (m, 2H).

[0167] Example 5 Preparation of (S)-4-(6-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-N-(piperidin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine (5)

[0168]

[0169] Step 1: Preparation of (S)-3-(2-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-6-methyl-1H-pyrrolo[2,3-b]pyridine-1-carboxylic acid tert-butyl ester (5a)

[0170] Compound 1c (100 mg, 0.17 mmol), trimethylcycloboroxane (25.2 mg, 0.20 mmol), Pd(dppf)Cl2 (14.52 mg, 0.02 mmol), and potassium carbonate (46.92 mg, 0.34 mmol) were dissolved in 1,4-dioxane (5 mL). Under nitrogen protection, the reaction mixture was heated to 100 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 80 mg of the title compound, yield: 81.6%.

[0171] LC-MS(ESI)m / z(M+H) + 577.5

[0172] Step 2: Preparation of (S)-4-(6-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-N-(piperidin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine (5)

[0173] Compound 5a (80 mg, 0.14 mmol) was dissolved in dichloromethane (5 mL), and then trifluoroacetic acid (2 mL) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 20 mg of the title compound, yield: 38.3%.

[0174] LC-MS(ESI)m / z(M+H) + 377.2

[0175] 1H NMR (400MHz, DMSO-d6) δ12.15 (s, 1H), 8.71 (d, J=8.1 Hz, 0.5H), 8.60-8.48 (m, 1.5H), 7.83 (s, 1H), 7.76 (s, 1H), 7.13-7.08 (m, 1H), 3.89 (s, 1H), 3.07 (d, J=11.9Hz, 1H) , 2.81 (d, J=12.3Hz, 1H), 2.55 (s, 3H), 2.48-2.41 (m, 2H), 1.99-1.92 (m, 1H), 1.70-1.61 (m, 1H), 1.48-1.42 (m, 2H).

[0176] Example 6 Preparation of (S)-N-(piperidin-3-yl)-4-(1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine (6)

[0177]

[0178] Step 1: Preparation of (S)-3-(2-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridine-1-carboxylic acid tert-butyl ester (6a)

[0179] Compound 1b (300 mg, 0.79 mmol), 1-(tert-butoxycarbonyl)-1H-pyrrolo[2,3-b]pyridine-3-boronic acid pinacol ester (357 mg, 1.04 mmol), Pd(dppf)Cl2 (58 mg, 0.08 mmol), and potassium carbonate (217.45 mg, 1.57 mmol) were dissolved in 1,4-dioxane (10 mL). Under nitrogen protection, the reaction system was heated to 100 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 200 mg of the title compound, yield: 45.1%.

[0180] LC-MS(ESI)m / z(M+H) + 563.1

[0181] Step 2: Preparation of (S)-N-(piperidin-3-yl)-4-(1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine (6)

[0182] Compound 6a (100 mg, 0.18 mmol) was dissolved in dichloromethane (5 mL), and then trifluoroacetic acid (2 mL) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 30 mg of the title compound, yield: 46.6%.

[0183] LC-MS(ESI)m / z(M+H) + 363.2

[0184] 1 H NMR (400MHz, DMSO-d6) δ12.35 (s, 1H), 8.82 (d, J=7.9Hz, 0.5H), 8.60-8.58 (m, 1.5H), 8.33 (dd, J=4.8, 1.5Hz, 1H), 7.92 (s, 1H), 7.86-7.83 (m, 1H), 7.25-7.18(m, 1H), 3.95(s, 1H), 3.12(d, J=12.0Hz, 1H), 2.90-2.85(m, 1H ), 2.52-2.43(m, 2H), 2.05-1.95(m, 1H), 1.70(s, 1H), 1.53-1.48(m, 2H).

[0185] Example 7 Preparation of (S)-4-(6-cyclopropyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-N-(piperidin-3-yl)-5-(trifluoromethyl)pyrimidin-2-amine (7)

[0186]

[0187] Using compound 1c and cyclopropylboronic acid as starting materials, the preparation process was the same as in Example 5, and the title compound was obtained.

[0188] LC-MS(ESI)m / z(M+H) + :403.1.

[0189] Biological evaluation

[0190] Experimental Example 1: HCC70 Cell Proliferation Inhibition Activity Test

[0191] 1. Experimental Objective

[0192] To evaluate the antiproliferative effect of the compound in HCC70 cells, the CCK-8 assay was used to measure cell viability and calculate the IC50 value. 50 .

[0193] 2. Test materials

[0194] 2.1. The compound to be tested:

[0195] Positive test drug: SY5609, purchased from MCE, batch number HY-138293;

[0196] Positive reagent: ICEC-0942, purchased from Shanghai Teber Chemical Technology Co., Ltd., batch number 2110809;

[0197] Test drugs: All compounds were prepared according to the examples in this application;

[0198] 2.2 Test Reagents and Instruments

[0199] CCK-8 test kit, Beyotime;

[0200] 1640 culture medium, Corning;

[0201] DMSO, Sigma;

[0202] Fetal bovine serum, Hyclone;

[0203] Microplate reader (BMG), POLARstar;

[0204] Pipettes, Eppendorf;

[0205] Countstar cell counter;

[0206] CO2 constant temperature incubator, Thermo;

[0207] Biosafety cabinet, Thermo;

[0208] Inverted microscope, Olympus;

[0209] 3. Test methods

[0210] 3.1 Compound Preparation

[0211] Preparation of 10mM compound stock solution: Dissolve the compound powder in 100% DMSO to prepare 10mM compound stock solutions.

[0212] 3.2 Test Methods

[0213] (1) Remove the 1640 complete culture medium from the cultured HCC70 cells and wash them twice with 1×PBS.

[0214] (2) Digest cells with 0.25% trypsin for 3-5 minutes until the cells become round and detach. Add 5 mL of 1640 complete culture medium to stop the digestion.

[0215] (3) Transfer the digested cells to a 15mL centrifuge tube and centrifuge at 1000rpm for 3min.

[0216] (4) Remove the supernatant, add 10 mL of 1640 complete culture medium to resuspend the cells, take 20 μL of cell suspension, add 20 μL of trypan blue, mix well, and count the cells using a cell counter. Select 3 fields of view to count and record the viable cell density, cell viability, and cell passage number.

[0217] (5) Dilute the cells with 1640 complete culture medium to a density of 1.11 × 10⁻⁶. 4 90 μL of cells were seeded into a 96-well plate, with 1000 cells per well.

[0218] (6) The inoculated cells were cultured overnight at 37°C in a CO2 constant temperature incubator.

[0219] (7) On the second day, take 10 mM of the compound stock solution and serially dilute it with 1640 complete culture medium containing 1% DMSO to make the compound concentrations 10, 3, 1, 0.3, 0.1, 0.03, 0.01, 0.003, and 0.001 μM. After the compound is prepared, take out the cell culture plate and add 10 μL of the diluted compound to the cells.

[0220] (8) The cells were cultured at 37°C in a CO2 constant temperature incubator for 3 days.

[0221] (9) On the third day of culture, remove the cell culture plate and add 10 μL of CCK-8 detection reagent to each well, and incubate for 4 hours. Read the values ​​using a microplate reader and calculate the inhibition rate.

[0222] 3.3 Data Analysis

[0223] Calculation formula

[0224]

[0225] Using the logarithm of concentration as the X-axis and the percentage inhibition rate as the Y-axis, dose-response curves were fitted using the log(inhibitor) vs. response-variable slop(four parameters) formula in the analysis software Graphpad Prism 9 to obtain the IC50 values ​​for each compound. 50 value.

[0226] 4. Test Results

[0227] The activity of the compounds tested in this application is shown in Table 1.

[0228] Table 1 Inhibitory activity of compounds tested in this application

[0229] Compound numbering <![CDATA[IC 50 (nM)]]> Compound numbering <![CDATA[IC 50 (nM)]]> 1 7.56 2 8.50 3 0.89 4 0.84 5 0.56 6 3.06 7 2.36 SY5609 18.13 ICEC-0942 361

[0230] As shown in Table 1, the test compound exhibits significant inhibitory activity against HCC70 cells.

[0231] Experimental Example 2: Assay on the Inhibitory Activity of MDA-MB-231 Cell Proliferation

[0232] 1. Experimental Objective

[0233] The antiproliferative effect of the compound in MDA-MB-231 cells was evaluated, and the IC50 was calculated using the Celltiter assay to assess cell viability. 50 .

[0234] 2. Test materials

[0235] 2.1. The compound to be tested:

[0236] Positive reagent: ICEC-0942, purchased from Shanghai Teber Chemical Technology Co., Ltd., batch number 2110809;

[0237] Positive test drug: SY5609, purchased from MCE, batch number HY-138293;

[0238] Test drugs: All compounds were prepared according to the examples in this application;

[0239] 2.2 Test Reagents and Instruments

[0240] Celltiter assay kit, Promega;

[0241] L15 culture medium, Coming;

[0242] DMSO, Sigma;

[0243] Fetal bovine serum, Hyclone;

[0244] Microplate reader (BMG), POLARstar;

[0245] Pipettes, Eppendorf;

[0246] Countstar cell counter;

[0247] CO2 constant temperature incubator, Thermo;

[0248] Biosafety cabinet, Thermo;

[0249] Inverted microscope, Olympus;

[0250] 3. Test methods

[0251] 3.1 Compound Preparation

[0252] Preparation of 10mM compound stock solution: Dissolve the compound powder in 100% DMSO to prepare 10mM compound stock solutions.

[0253] 3.2 Test Methods

[0254] (1) Remove L15 culture medium from the cultured MDA-MB-231 cells and wash them twice with 1×PBS.

[0255] (2) Digest cells with 0.25% trypsin for 3-5 minutes until the cells become round and detach. Add 5 mL of L15 complete culture medium to stop the digestion.

[0256] (3) Transfer the digested cells to a 15mL centrifuge tube and centrifuge at 1000rpm for 3min.

[0257] (4) Remove the supernatant, add 10 mL of L15 complete culture medium to resuspend the cells, take 20 μL of cell suspension, add 20 μL of trypan blue, mix well, and count the cells using a cell counter. Select 3 fields of view to count and record the viable cell density, cell viability, and cell passage number.

[0258] (5) Dilute the cells with L15 complete culture medium to a density of 1.11 × 10⁻⁶. 4 90 μL of cells were seeded into a 96-well plate, with 1000 cells per well.

[0259] (6) The inoculated cells were cultured overnight at 37°C in a CO2 constant temperature incubator.

[0260] (7) On the second day, take 10 mM of the compound stock solution and serially dilute it with L15 complete culture medium containing 1% DMSO to make the compound concentrations 30, 10, 3, 1, 0.3, 0.1, 0.03, 0.01, and 0.003 μM. After the compound is prepared, take out the cell culture plate and add 10 μL of the diluted compound to the cells.

[0261] (8) The cells were cultured at 37°C in a CO2 constant temperature incubator for 3 days.

[0262] (9) On the third day of culture, remove the cell culture plate and add 50 μL of Celltiter assay reagent to each well. Lyse the cells on a horizontal shaker in the dark for 10 min, and aspirate 20 μL to read the values ​​in the 384-well white plate.

[0263] 3.3 Data Analysis

[0264] Calculation formula

[0265]

[0266] Using the logarithm of concentration as the X-axis and the percentage inhibition rate as the Y-axis, dose-response curves were fitted using the log(inhibitor) vs. response-variable slop(four parameters) formula in the analysis software Graphpad Prism 9 to obtain the IC50 values ​​for each compound. 50 value.

[0267] 4. Test Results

[0268] The activity of the compounds tested in this application is shown in Table 2.

[0269] Table 2 Inhibitory activity of compounds tested in this application

[0270]

[0271]

[0272] As shown in Table 2, the test compound exhibits significant inhibitory activity against MDA-MB-231 cells.

[0273] Experimental Example 3: CDK7 Enzyme Activity Assay

[0274] 1. Experimental Objective

[0275] This experiment reflects kinase activity by detecting the amount of ADP generated in the kinase reaction. A three-step method was used to detect the inhibitory activity of compounds on the CDK7 enzyme. The first step involved incubating CDK7 / CyclinH1 / MNAT1, different concentrations of the compound, substrate (MBP), and ATP at room temperature for a fixed time. The second step involved adding ADPglo assay reagent to terminate the enzymatic reaction, followed by further incubation at room temperature for a fixed time. The third step involved adding Kinase Detection Reagent, incubating at room temperature for a fixed time, and then taking readings. Continuous readings were performed using the Lumi module of a BMG microplate reader to evaluate the effect of the test compounds on CDK7 enzyme activity. Simultaneously, the IC50 of the test compounds on CDK7 enzyme was calculated using the inhibition rate. 50 value.

[0276] 2. Test materials

[0277] 2.1. The compound to be tested:

[0278] Positive reagent: ICEC-0942, purchased from Shanghai Teber Chemical Technology Co., Ltd., batch number 2110809:

[0279] Test drugs: All compounds were prepared according to the examples in this application.

[0280] 2.2 Test Reagents and Instruments

[0281] CDK7 / CyclinH1 / MNAT1, Promega;

[0282] ADPglo, Promega

[0283] DMSO, Sigma;

[0284] OptiPlate-384, Thermo;

[0285] Microplate reader (BMG), POLARstar;

[0286] Pipettes, Eppendorf;

[0287] 3. Test methods

[0288] 3.1 Preparation of test compounds

[0289] Preparation of 10mM compound stock solution: Dissolve the compound powder in 100% DMSO to prepare 10mM compound stock solutions.

[0290] 3.2 Enzyme Reaction Process

[0291] (1) Prepare 1×PBS buffer.

[0292] (2) Compound concentration preparation: Compound IC 50 The final concentration for testing started at 10 μM, diluted 5-fold, resulting in 6 concentrations, with each concentration set up for single / duplicate detection. 1 μL of the diluted compound solution was added to each well, and the solution was diluted 5-fold in a 384-well plate to the corresponding final concentration. For the negative control, no compound or CDK7 / CyclinH1 / MNAT1 was added, and the remaining volume was made up with 1×PBS buffer. For the positive control, no compound was added, and the remaining volume was made up with 1×PBS buffer.

[0293] (3) Prepare a CDK7 / CyclinH1 / MNAT1 enzyme (15 ng / well) solution using 1×PBS buffer, and add 2 μL of the solution to each well.

[0294] (4) Prepare a mixed solution of substrate and ATP (0.33 ng / well) using 1×PBS buffer, and add 2 μL of the mixed solution to each well.

[0295] (Start the reaction at 5J and incubate at room temperature for 60 minutes.)

[0296] (6) Add 5 μL of ADPglo reagent to each well and incubate at room temperature for 50 min.

[0297] (7) Add 10 μL of detection solution to each well and incubate at room temperature for 30 min.

[0298] (8) Read the signal value using the Lumi module of the BMG microplate reader, and read continuously (1 min / time) for a total of 30 times.

[0299] 3.3 Data Analysis

[0300] Calculation formula

[0301]

[0302] Plotting the logarithm of concentration on the X-axis and the percentage inhibition rate on the Y-axis, dose-response curves were fitted using the log(inhibitor) vs. response-variable slop(four parameters) formula in Graphpad Prism 9 software to derive the IC50 of each compound on enzyme activity. 50 value.

[0303] 4. Test Results

[0304] The inhibitory activity of the compounds in this application against CDK7 enzyme is shown in Table 1.

[0305] Table 3. Inhibitory activity of the compounds tested in this application against CDK7 enzyme.

[0306] Compound numbering <![CDATA[IC 50 (nM)]]> Compound numbering <![CDATA[IC 50 (nM)]]> 4 3.7 5 3.2 6 3.3 ICEC-0942 21

[0307] As can be seen from the test data of CDK7 enzyme inhibitory activity of the test compounds shown in Table 3, the compounds of this application have significant CDK7 enzyme inhibitory activity.

[0308] Experimental Example 4: Pharmacokinetic Study in Rats

[0309] 1. Experimental Objective

[0310] Using SD rats as test animals, the plasma concentrations of the compound of this application at different time points after oral administration were determined by LC-MS / MS. The pharmacokinetic behavior of the compound of this application in rats was studied to evaluate its pharmacokinetic characteristics.

[0311] 2. Test materials

[0312] 2.1. The compound to be tested:

[0313] Positive reagent: ICEC-0942, purchased from Shanghai Teber Chemical Technology Co., Ltd., batch number 2110809;

[0314] Test drugs: All compounds were prepared according to the examples in this application;

[0315] 2.2. Test Instruments:

[0316] Shimadzu LC-30AABAPI4500 tandem mass spectrometer, vacuum blood collection tubes, blood collection needles, filter paper, syringes, etc.

[0317] 2.3 Experimental Animals

[0318] Male SD rats, weighing 180-220g, were used in groups of 3. After purchase, the animals were housed in an animal facility for at least 3 days to acclimatize, and were used in the experiment only after passing quarantine.

[0319] 3. Test methods

[0320] 3.1 Grouping: The rats were randomly grouped according to Table 4. After grouping, there was no statistically significant difference in body weight among the different groups of SD rats.

[0321] Table 4. Trial Groups and Dosing Regimens

[0322]

[0323] 3.2 Blood sample collection and testing:

[0324] According to Table 4, each group was administered the corresponding test drug by gavage. Before administration, and 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after administration, 200 μL of blood was collected through the orbital sinus and placed in EDTA-K2 anticoagulant tubes. The plasma was separated into centrifuge tubes at 5000 rpm for 10 min and then frozen at -80℃.

[0325] 3.3 Analytical Methods

[0326] Take out the plasma at each time point stored at -80℃, add acetonitrile, vortex at 1500 rpm for 2 min, centrifuge for 15 min (3500 rpm), and take out the supernatant of the solution for LC-MS / MS analysis.

[0327] 4. Calculation of pharmacokinetic parameters:

[0328] The pharmacokinetic behavior of the test compounds was fitted using a non-compartmental model, and the main pharmacokinetic parameters (T0, T ...) were calculated using DAS 3.31 software. 1 / 2 T max C max AUC last wait).

[0329] 5. Test Results:

[0330] Table 5 Pharmacokinetic parameters of compounds in the examples

[0331] Compound numbering <![CDATA[C max (ng / mL)]]> <![CDATA[T max (h)]]> <![CDATA[T 1 / 2 (h)]]> <![CDATA[AUC last (ng / mL*h)]]> ICEC-0942 23.7±17.4 1.7±0.6 13.5±4.8 103.6±27.8 Compound 4 286.0±87.9 1.3±0.9 5.3±0.6 1396.7±276.6 Compound 5 140.7±6.1 2.7±1.1 5.4±0.9 692.1±138.8 Compound 6 83.2±19.8 2.7±1.1 3.6±0.5 440.6±153.0

[0332] As can be seen from the test results shown in Table 5, compared with the positive group, compounds 4, 5 and 6 are superior to ICEC-0942 in terms of plasma exposure, indicating that the pharmacokinetic properties of the compounds in this application are significantly improved compared with ICEC-0942.

[0333] Experimental Example 5: In vivo efficacy trial in a subcutaneous xenograft tumor model of HCC70 cells

[0334] 1. Test Methods

[0335] 1.1 Cell Culture

[0336] Cultured in RPMI 1640 medium containing 20% ​​fetal bovine serum (FBS) at 37°C and 5% CO2.

[0337] 1.2 Compounds:

[0338] Solvent: 10% DMSO+90% (SBE-β-CD in saline);

[0339] Test drug: Compound 5;

[0340] Positive test results: ICEC-0942 and SY5609.

[0341] 1.3 Experimental Procedure

[0342] Balb / c nude mice, female, 6-8 weeks old, weighing approximately 18-22 grams, were subcutaneously injected with 0.1 mL (1×10⁻⁶) on the right side of each mouse. 7 HCC70 cells (cells + matrix gel). When the average tumor volume reached 100-200 mm³, mice were divided into groups and administered the drug. The dosage and administration method are shown in Table 6 below. Tumor volume was measured twice a week, and the mouse's body weight, tumor length and width were recorded at each measurement. Tumor volume = length × width × width / 2 mm. 3 When the average tumor volume in the solvent group reached over 3000 mm³, administration was terminated, and mice in each group were sacrificed. Tumor masses were dissected, and the tumor weight and volume of each mouse were weighed. The differences in average tumor volume between the positive control group (ICEC-0942 and SY5609), the test compound group (compound 5), and the solvent group were compared. The tumor-inhibiting effect of the compounds was evaluated using TGI (%), which reflects the tumor growth inhibition rate.

[0343] TGI (%) = [1 - (average tumor volume of compound in the current test - average tumor volume of compound at the beginning) / (average tumor volume of solvent group in the current test - average tumor volume of solvent group at the beginning)] × 100%

[0344] 2. Test Results

[0345] Table 6. In vivo tumor suppression experiment data

[0346] Group Number of animals Administration method Dosage (mg / Kg) Number of days of medication TGI (%) solvent group 8 qd, po / 21 / Compound 4 8 qd, po 2.5 21 68.5 Compound 5 8 qd, po 2.5 21 74.1 Compound 6 8 qd, po 2.5 21 71.0 ICEC-0942 8 qd, po 100 21 36.8 SY5609 8 qd, po 5.0 21 74.6

[0347] 3. Experimental Conclusions

[0348] The above experimental results show that the compound of this application exhibits good in vivo efficacy in a subcutaneous xenograft tumor model of breast cancer HCC70 cells, with significant tumor-suppressing activity (TGI > 60%). This tumor-suppressing effect is superior to that of the clinical investigational compounds ICEC-0942 and SY5609, and the test animals showed good tolerance to the compound, with no decrease in mouse body weight after administration. Therefore, the compound of this application can demonstrate good in vivo tumor-suppressing effects.

Claims

1. A compound, its stereoisomer, or a pharmaceutically acceptable salt thereof, said compound being selected from:

2. A method for preparing the compound according to claim 1, comprising: (1) Compound I-1 and compound I-2 were subjected to a substitution reaction to obtain compound I-3; (2) Compound I-3 and compound I-4 were coupled together to obtain compound I-5; (3) Compound I-5 was deprotected to obtain a compound having the structure of formula I; Where LG represents the leaving group and PG represents the protecting group. for R 1 Selected from hydrogen, chlorine, methyl, cyclopropyl or R 2 Selected from methyl, ethyl, or isopropyl; R 3 R 4 Selected independently from hydrogen, m is selected from 0.

3. The preparation method according to claim 2, characterized in that, The leaving group is selected from halogen atoms, methanesulfonyloxy groups, or p-toluenesulfonyloxy groups; the protecting group is selected from 2-(trimethylsilyl)ethoxymethyl or tert-butyloxycarbonyl.

4. A pharmaceutical composition, characterized in that, It comprises the compound of claim 1, its stereoisomers or pharmaceutically acceptable salts thereof, and pharmaceutically acceptable excipients.

5. Use of the compound, its stereoisomer, or a pharmaceutically acceptable salt thereof, as described in any one of claims 1, in the preparation of a medicament for the prevention or treatment of cyclin-dependent kinase 7 (CDK7) mediated diseases or conditions.

6. The use as described in claim 5, characterized in that, The CDK7-mediated diseases or conditions are selected from tumors or cancers.

7. The use as described in claim 6, characterized in that, The diseases mentioned are selected from breast cancer, pancreatic cancer, ovarian cancer, colorectal cancer, lung cancer, prostate cancer, lymphoma, malignant sarcoma, cervical cancer, oral cancer, brain cancer, stomach cancer, liver cancer, skin cancer, bone cancer, kidney cancer, bladder cancer, fallopian tube tumors, peritoneal tumors, melanoma, glioma, glioblastoma, papillary malignant tumors, head and neck tumors, myeloma, or leukemia.

8. Use of the compound, its stereoisomer, or a pharmaceutically acceptable salt thereof, as described in any one of claims 1, in the preparation of a CDK7 inhibitor.

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