Selective CDK12 / 13 inhibitor and application thereof

Abstract

The invention belongs to the field of medical chemistry, relates to a compound serving as a CDK12 / 13 inhibitor and application of the compound, and particularly provides a compound shown in a formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug of the compound. Processes for their preparation, pharmaceutical compositions containing these compounds and the use of these compounds or compositions for the treatment of diseases associated with CDK12 / 13. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and specifically relates to compounds or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs as selective CDK12 / 13 inhibitors, their preparation methods, and pharmaceutical compositions containing these compounds and the use of these compounds or compositions for treating diseases related to CDK12 / 13. Background Art

[0002] Members of the cyclin-dependent kinase (CDK) family play key regulatory roles in cell proliferation. There are more than 20 known mammalian CDKs. CDK12 and CDK13 were identified in a cDNA screen for cell cycle regulators.

[0003] CDK12 (also known as CrkRS, CRK7) is a protein composed of 1490 amino acids with a molecular weight of 164kDa. Its protein structure can be divided into three main domains: N-terminal domain, kinase domain and C-terminal domain. In CDK12, the RS domain at the N-terminus is very conserved and plays an important role in mRNA splicing. CDK12 has multiple functions, such as phosphorylating the CTD region of RNA polymerase Pol II, which can promote the elongation of transcription; can interact with RNA processing factors to regulate the RNA shearing process; mediate the phosphorylation of transcribed RNA polymerase II, regulate the processing of mRNA 3' end; and regulate the polyadenylation and transcription of introns (Cancer Res., 2021, 81, 18-26).

[0004] The CDK that is most closely related to CDK12 is CDK13 (also known as CDC2L, CDC2L5, CHED). CDK13 is the largest CDK, consisting of 1512 amino acids. Its kinase domain is 92% similar to the kinase domain of CDK12, but there are significant differences in their structures outside this region. Similar to CDK12, CDK13 can also form heterodimers with CyclinK to play a kinase role. It can also catalyze the phosphorylation of Ser2 and Ser5 in the CTD region of RNA polymerase Pol II like CDK12 to play subsequent functions. Studies have shown (Cell Reports, 2016, 14, 320-331) that in addition to this common biochemical function, CDK12 and CDK13 regulate different gene clusters of expression. In addition, CDK13 can also regulate the expression of snRNA and snoRNA.

[0005] Studies have found that CDK12 and CDK13 are closely related to a variety of cancers and pathways, and mutations, amplifications, deletions and fusions of the CDK12 / 13 genome have been found in cancer. Studies have shown that CDK12 / 13 gene mutations and overexpression are widely present in colorectal cancer, ovarian cancer, breast cancer, esophageal cancer, gastric cancer, endometrial cancer, bladder cancer, pancreatic cancer and other cancers, and patients with high expression of CDK12 / 13 in a variety of tumors have a poor prognosis.

[0006] Most of the existing small molecule CDK inhibitors are pan-CDK inhibitors. This type of inhibitor usually has defects such as insufficient efficacy, high toxicity, and a narrow safety window. Studies have shown that CDK1 and CDK7 play an important role in cell cycle regulation and transcription in normal cells. Therefore, the development of a new generation of selective CDK inhibitors has certain clinical application significance for improving efficacy and reducing toxic side effects. Given that CDK12 / 13 plays an important role in the occurrence and development of multiple tumors such as gastric cancer, colorectal cancer, and breast cancer, the development of selective inhibitors for CDK12 / 13 should have good clinical application prospects. Summary of the invention

[0007] One object of the present invention is to provide a class of compounds having CDK12 / 13 inhibitory activity represented by general formula (I), general formula (Ia), general formula (II) or general formula (III) or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs.

[0008] Another object of the present invention is to provide a method for preparing the compounds of the general formula (I), general formula (Ia), general formula (II) or general formula (III) of the present invention or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs.

[0009] Another object of the present invention is to provide a composition comprising a compound of the present invention of general formula (I), general formula (Ia), general formula (II) or general formula (III) or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug and a pharmaceutically acceptable carrier, as well as a composition comprising a compound of the present invention of general formula (I), general formula (Ia), general formula (II) or general formula (III) or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug and one or more other drugs.

[0010] Another object of the present invention is to provide a method for treating diseases associated with CDK12 / 13 using the compounds of the present invention of general formula (I), general formula (Ia), general formula (II) or general formula (III) or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs, as well as use of the compounds of the present invention of general formula (I), general formula (Ia), general formula (II) or general formula (III) or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs in the preparation of drugs for treating diseases associated with CDK12 / 13.

[0011] In view of the above-mentioned invention object, the present invention provides the following technical solutions:

[0012] In a first aspect, the present invention provides a compound represented by general formula (I) or an isomer, a pharmaceutically acceptable salt, a solvate, a crystal or a prodrug thereof,

[0013]

[0014] in,

[0015] X1 and X2 are each independently selected from -CH- and -N-;

[0016] R1, R2 and R3 are each independently selected from hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, oxygen, sulfur, thiol, alkylthio, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylalkylthio, heterocyclylalkylthio, arylalkylthio and heteroarylalkylthio, which are optionally substituted with one or more alkyl, alkoxy, alkylamino, halogen, hydroxy, amino, nitro, cyano, haloalkyl, alkylacyl, aminoacyl or alkylaminoacyl; or R1 and R2 together with the C atoms to which they are attached form cycloalkyl, heterocyclyl, aryl, heteroaryl, which are optionally substituted with one or more alkyl, alkoxy, alkylamino, halogen, hydroxy, amino, nitro, cyano, haloalkyl, alkylacyl, aminoacyl or alkylaminoacyl;

[0017] R5 and R6 are each independently selected from hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, alkylthio, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino and oxo;

[0018] R7 and R8 are each independently selected from hydrogen, halogen, hydroxy, carbonyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, alkylcarbonyl, alkenylcarbonyl, alkynylcarbonyl, optionally substituted with one or more alkyl, alkenyl, alkynyl, alkoxy, alkylamino, halogen, hydroxy, amino, haloalkyl, alkylacyl, aminoacyl or alkylaminoacyl; and

[0019] is a single bond or a double bond, where When it is a double bond, R4 does not exist; When it is a single bond, R4 is selected from alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, alkylthio, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl and dialkylamino, and R3 is selected from hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, oxygen, sulfur, mercapto, alkylthio, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino, oxo, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylalkylthio, heterocyclylalkylthio, arylalkylthio and heteroarylalkylthio.

[0020] In some embodiments, the compound of the general formula (I) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug has the following structure of the general formula (Ia):

[0021]

[0022] Among them, R1, R2, R3, R4, R5, R6, R7 and As defined in general formula (I).

[0023] In a second aspect, the present invention provides a compound represented by general formula (II) or an isomer, a pharmaceutically acceptable salt, a solvate, a crystal or a prodrug thereof,

[0024]

[0025] Among them, X1, X2, R1, R2, R3, R4, R5, R6, R7, R8 and As defined in general formula (I).

[0026] In a third aspect, the present invention provides a compound represented by general formula (III) or an isomer, a pharmaceutically acceptable salt, a solvate, a crystal or a prodrug thereof,

[0027]

[0028] Among them, X1, X2, R1, R2, R3, R4, R5, R6, R7, R8 and As defined in formula (I) and formula (II).

[0029] In some embodiments, according to the compound of general formula (I), general formula (II) or general formula (III) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, X1 is -N-, X2 is -CH-, R7 is selected from hydrogen, halogen, hydroxyl, carbonyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, alkylcarbonyl, alkenylcarbonyl, alkynylcarbonyl, which is optionally substituted with one or more alkyl, alkenyl, alkynyl, alkoxy, alkylamino, halogen, hydroxyl, amino, haloalkyl, alkylacyl, aminoacyl and alkylaminoacyl, and R8 is hydrogen. In other embodiments, according to the compound of general formula (I), general formula (II) or general formula (III) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, X1 is -CH-, X2 is -N-, R7 is hydrogen, and R8 is selected from hydrogen, halogen, hydroxyl, carbonyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, alkylcarbonyl, alkenylcarbonyl, alkynylcarbonyl, which is optionally substituted with one or more alkyl, alkenyl, alkynyl, alkoxy, alkylamino, halogen, hydroxyl, amino, haloalkyl, alkylacyl, aminoacyl or alkylaminoacyl.

[0030] In some preferred embodiments, the compound of the present invention is a compound of formula (I), formula (Ia), formula (II) or formula (III) or an isomer, a pharmaceutically acceptable salt, a solvate, a crystal or a prodrug thereof, wherein R1, R2 and R3 are each independently selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, oxygen, sulfur, mercapto, C 1-6 Alkylthio, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, di-C 1-6 Alkylamino, C 3-8 Cycloalkyl, C 3-8 Heterocyclic group, C 6-12 Aryl, C 5-12 Heteroaryl, C 3-8 Cycloalkyl C1-6 Alkylthio, C 3-8 Heterocyclyl C 1-6 Alkylthio, C 6-12 Aryl C 1-6 Alkylthio and C 5-12 Heteroaryl C 1-6 Alkylthio, which is optionally substituted with one or more C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, halogen, hydroxy, amino, nitro, cyano, halo C 1-3 Alkyl, C 1-3 Alkyl acyl, amino acyl or C 1-3 Alkylaminoacyl substitution.

[0031] In some specific embodiments, the compound of the present invention is a compound of formula (I), formula (Ia), formula (II) or formula (III) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein R1, R2 and R3 are each independently selected from It is optionally replaced by one or more C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, halogen, hydroxy, amino, nitro, cyano, halo C 1-6 Alkyl, C 1-6 Alkyl acyl, amino acyl or C 1-6 Alkylaminoacyl substitution.

[0032] In some embodiments, the compound of the present invention is a compound of formula (I), formula (Ia), formula (II) or formula (III) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein:

[0033] R1, R2 and R3 are each independently selected from hydrogen, halogen, hydroxyl, C 1-3 Alkyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, nitro, carboxyl, cyano, amino, oxygen, sulfur, mercapto, C 1-3 Alkylthio, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl, C 1-3 Alkylaminoacyl, di-C 1-3 Alkylamino, C 3-8 Cycloalkyl, C 3-8 Heterocyclic group, C6-12 Aryl, C 5-12 Heteroaryl, C 3-8 Cycloalkyl C 1-3 Alkylthio, C 3-8 Heterocyclyl C 1-3 Alkylthio, C 6-12 Aryl C 1-3 Alkylthio and C 5-12 Heteroaryl C 1-3 Alkylthio, which is optionally substituted with one or more C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkylamino, halogen, hydroxy, amino, nitro, cyano, halo C 1-3 Alkyl, C 1-3 Alkyl acyl, amino acyl or C 1-3 Alkylaminoacyl substitution;

[0034] Further preferably, R1, R2 and R3 are each independently selected from hydrogen, halogen, hydroxy, methyl, ethyl, propyl, isopropyl, isobutyl, trifluoromethyl, trifluoroethyl, hydroxy 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, nitro, carboxyl, cyano, amino, oxygen, sulfur, mercapto, C 1-3 Alkylthio, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl, C 1-3 Alkylaminoacyl, di-C 1-3 Alkylamino, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxa-C 3-6 Cyclic group, nitrogen C 3-6 Cyclic group, C 6-12 Aryl, Aza C 5-12 Heteroaryl, OxaC 5-12 Heteroaryl, nitrogen, oxygen, carbonyl 5-12 Heteroaryl, C 3-6 Cycloalkyl C 1-3 Alkylthio, oxa-C 3-6 Heterocyclyl C 1-3 Alkylthio, AzaC 3-6 Heterocyclyl C 1-3 Alkylthio, nitrogen oxide C 3-6 Heterocyclyl C 1-3 Alkylthio, C 6-12 Aryl C 1-3 Alkylthio, AzaC 5-12 Heteroaryl C 1-3 Alkylthio, oxa-C 5-12 Heteroaryl C1-3 Alkylthio, nitrogen oxide C 5-12 Heteroaryl C 1-3 Alkylthio, which is optionally substituted by one or more hydroxy, methyl, ethyl, propyl, isopropyl, isobutyl, trifluoromethyl, trifluoroethyl, C 1-3 Alkoxy, C 1-3 Alkylamino, halogen, hydroxy, amino, nitro, cyano, halo C 1-3 Alkyl, C 1-3 Alkyl acyl, amino acyl or C 1-3 Alkylaminoacyl substitution.

[0035] In some specific embodiments, according to the compounds of formula (I), formula (Ia), formula (II) or formula (III) of the present invention or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs, R1 is selected from hydrogen, R2 is selected from hydrogen, fluorine, chlorine, bromine, trifluoromethyl and C 1-6 Alkyl, R3 is selected from hydrogen, fluorine, chlorine, bromine, trifluoromethyl, C 1-6 Alkyl and oxo groups, R4 is selected from hydrogen and C 1-6 In other specific embodiments, according to the compound of general formula (I), general formula (Ia), general formula (II) or general formula (III) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, Selected from R4 is selected from hydrogen and C 1-6 alkyl.

[0036] In some embodiments, the compound of the present invention is a compound of formula (I), formula (Ia), formula (II) or formula (III) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein:

[0037] R5 and R6 are each independently selected from hydrogen, halogen, hydroxy, alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, C 1-6 Alkylthio, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, di-C 1-6 Alkylamino and oxo groups;

[0038] Further preferably, R5 and R6 are each independently selected from hydrogen, halogen, hydroxyl, C 1-3 Alkyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, nitro, carboxyl, cyano, amino, C 1-3 Alkylthio, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl, C 1-3 Alkylaminoacyl, di-C 1-3 Alkylamino and oxo groups;

[0039] More preferably, R5 and R6 are each independently selected from hydrogen, halogen, hydroxy, methyl, ethyl, propyl, isopropyl, isobutyl, trifluoromethyl, trifluoroethyl, hydroxy 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, nitro, carboxyl, cyano, amino, oxygen, sulfur, mercapto, C 1-3 Alkylthio, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl, C 1-3 Alkylaminoacyl, di-C 1-3 Alkylamino and oxo groups.

[0040] In some specific embodiments, according to the compounds of general formula (I), general formula (Ia), general formula (II) or general formula (III) of the present invention or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs, R5 and R6 are each independently selected from hydrogen, methyl, ethyl, propyl, fluorine, chlorine and bromine.

[0041] In some embodiments, the compound of the present invention is a compound of formula (I), formula (II) or formula (III) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein:

[0042] R7 and R8 are each independently selected from hydrogen, halogen, hydroxyl, carbonyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, C 1-6Alkylcarbonyl, C 2-6 Alkenylcarbonyl, C 2-6 Alkynylcarbonyl, which is optionally substituted with one or more C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Alkylamino, halogen, hydroxy, amino, halo C 1-6 Alkyl, C 1-6 Alkyl acyl, amino acyl or C 1-6 Alkylaminoacyl substitution;

[0043] Further preferably, R7 and R8 are each independently selected from hydrogen, halogen, hydroxyl, carbonyl, C 1-3 Alkyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, nitro, carboxyl, cyano, amino, C 1-3 Alkylcarbonyl, C 2-4 Alkenylcarbonyl, C 2-4 Alkynylcarbonyl, which is optionally substituted with one or more C 1-3 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-3 Alkoxy, C 1-3 Alkylamino, halogen, hydroxy, amino, halo C 1-3 Alkyl, C 1-3 Alkyl acyl, amino acyl or C 1-3 Alkylaminoacyl substitution;

[0044] More preferably, R7 and R8 are each independently selected from hydrogen, halogen, hydroxy, carbonyl, methyl, ethyl, propyl, isopropyl, isobutyl, trifluoromethyl, trifluoroethyl, hydroxy 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, nitro, carboxyl, cyano, amino, C 1-3 Alkylcarbonyl, C 2-4 Alkenylcarbonyl, C 2-4 Alkynylcarbonyl, which is optionally substituted with one or more C 1-3 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-3 Alkoxy, C 1-3 Alkylamino, halogen, hydroxy, amino, halo C 1-3 Alkyl, C 1-3 Alkyl acyl, amino acyl or C1-3 Alkylaminoacyl substitution.

[0045] In some specific embodiments, according to the compounds of formula (I), (II) or (III) of the present invention or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs, R7 and R8 are each independently selected from hydrogen,

[0046] In some specific embodiments, according to the compound of general formula (I), general formula (II) or general formula (III) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, X1 is -N-, X2 is -CH-, R7 is In other specific embodiments, according to the compound of general formula (I), general formula (II) or general formula (III) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, X1 is -CH-, X2 is -N-, R7 is hydrogen, and R8 is

[0047] In some specific embodiments, according to the compound of general formula (I), general formula (Ia), general formula (II) or general formula (III) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, is a double bond, and R4 does not exist. In other specific embodiments, according to the compound of general formula (I), general formula (Ia), general formula (II) or general formula (III) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, is a single bond, R4 is selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, C 1-6 Alkylthio, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl and di-C 1-6 Alkylamino, R3 is selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, oxygen, sulfur, mercapto, C1-6 Alkylthio, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, di-C 1-6 Alkylamino, oxo, C 3-8 Alkyl, C 3-8 Heterocyclic group, C 6-12 Aryl, C 5-12 Heteroaryl, C 3-8 Cycloalkyl C 1-6 Alkylthio, C 3-8 Heterocyclyl C 1-6 Alkylthio, C 6-12 Aryl C 1-6 Alkylthio and C 5-12 Heteroaryl C 1-6 Alkylthio, wherein the heteroatom is selected from N, O, P and S.

[0048] The present invention provides the following specific compounds or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs:

[0049]

[0050]

[0051] In a fourth aspect, the present invention provides a pharmaceutical composition comprising a compound represented by the general formula (I), general formula (Ia), general formula (II) or general formula (III) of the present invention or an isomer, a pharmaceutically acceptable salt, a solvate, a crystal or a prodrug thereof.

[0052] In some embodiments, the present invention provides compounds of the general formula (I), general formula (Ia), general formula (II) or general formula (III) of the present invention or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs, and pharmaceutical compositions comprising compounds of the general formula (I), general formula (Ia), general formula (II) or general formula (III) of the present invention or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs, wherein the compounds or pharmaceutical compositions are used to treat diseases associated with CDK12 / 13.

[0053] In some embodiments, the present invention provides a pharmaceutical composition comprising a compound represented by formula (I), formula (Ia), formula (II) or formula (III) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug and a pharmaceutically acceptable carrier.

[0054] The compound shown in the general formula (I), general formula (Ia), general formula (II) or general formula (III) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug can be mixed with a pharmaceutically acceptable carrier, diluent or excipient to prepare a pharmaceutical preparation suitable for oral or parenteral administration. The method of administration includes, but is not limited to intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal and oral routes. The preparation can be administered by any route, such as by infusion or push injection, by a route of absorption through the epithelium or skin mucosa (such as oral mucosa or rectum, etc.). Administration can be systemic or local. Examples of oral administration preparations include solid or liquid dosage forms, specifically, tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions, etc. The preparation can be prepared by methods known in the art and include carriers, diluents or excipients conventionally used in the field of pharmaceutical preparations.

[0055] In a fifth aspect, the present invention provides a method for treating diseases associated with CDK12 / 13 using a compound represented by general formula (I), general formula (Ia), general formula (II) or general formula (III) of the present invention or its isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs, or a pharmaceutical composition comprising the same, as well as use in the preparation of drugs for treating diseases associated with CDK12 / 13.

[0056] In some embodiments, the present invention provides compounds represented by general formula (I), general formula (Ia), general formula (II) or general formula (III) of the present invention or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs, or pharmaceutical compositions comprising the same for treating diseases associated with abnormal activity of CDK12 / 13.

[0057] In some preferred embodiments, the present invention provides a compound represented by general formula (I), general formula (Ia), general formula (II) or general formula (III) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, or a pharmaceutical composition comprising the same for use in a method for treating a disease associated with CDK12 / 13 and in the preparation of a drug for treating a disease associated with CDK12 / 13, wherein the disease associated with CDK12 / 13 includes but is not limited to proliferative diseases, metabolic diseases, inflammatory diseases, autoimmune diseases and infectious diseases. In some embodiments, the proliferative disease associated with CDK12 / 13 described in the present invention is cancer.

[0058] In some embodiments, the present invention provides a method for treating a CDK12 / 13-mediated disease of a compound of formula I, formula (II) or formula (III) of the present invention, or an isomer thereof, a pharmaceutically acceptable salt, solvate, crystal or prodrug or pharmaceutical composition thereof, comprising administering a therapeutically effective amount of a compound of formula I, formula (II) or formula (III) of the present invention, or an isomer thereof, a pharmaceutically acceptable salt, solvate, crystal or prodrug or pharmaceutical composition thereof. In some embodiments, the present invention provides a method for treating a proliferative disease, comprising administering a therapeutically effective amount of a compound of formula I, formula (II) or formula (III) of the present invention, or an isomer thereof, a pharmaceutically acceptable salt, solvate, crystal or prodrug thereof. In some embodiments, the present invention provides a method for inhibiting tumor cell growth and / or metastasis, comprising administering a therapeutically effective amount of a compound of formula I, formula (II) or formula (III) of the present invention, or an isomer thereof, a pharmaceutically acceptable salt, solvate, crystal or prodrug thereof.

[0059] In some embodiments, the diseases associated with CDK12 / 13 described in the present invention include but are not limited to: acoustic neuroma, adenocarcinoma, adrenal cancer, anal cancer, angiosarcoma (e.g., lymphangiosarcoma, lymphangioendothelial sarcoma, angiosarcoma), adnexal cancer, benign monoclonal gammopathy, bile cancer (e.g., bile duct cancer), bladder cancer, breast cancer (e.g., breast adenocarcinoma, breast papillary carcinoma, breast cancer, medullary breast cancer, triple-negative breast cancer), brain cancer (e.g., meningioma; glioma, such as astrocytoma, oligodendroglioma; medulloblastoma), bronchial carcinoma, carcinoid tumor, cervical cancer (e.g., cervical adenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma), epithelial cancer, ependymoma, endothelial sarcoma (e.g., Kaposi's sarcoma (Kaposi's sarcoma), multiple idiopathic hemorrhagic sarcomas), endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophageal cancer (e.g., esophageal adenocarcinoma, Barrett's adenocarinoma), Ewing sarcoma (e.g., sarcoma), eye cancer (e.g., intraocular melanoma, retinoblastoma), familial eosinophilia, gallbladder cancer, stomach cancer (e.g., gastric adenocarcinoma), gastrointestinal stromal tumor (GIST), head and neck cancer (e.g., head and neck squamous cell carcinoma), oral cancer (e.g., oral squamous cell carcinoma (OSCC), pharyngeal cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal carcinoma, oropharyngeal cancer)), hematopoietic cancer (e.g., leukemia such as acute lymphocytic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acute myeloid leukemia (AML) (e.g., B-cell AML, T-cell AML), chronic myeloid leukemia (CML) (e.g., B-cell CML, T-cell CML) and chronic lymphocytic leukemia (CLL) (e.g., B-cell CL L, T-cell CLL); lymphomas such as Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and non-Hodgkin lymphoma (NHL) (e.g., B-cell NHL such as diffuse large cell lymphoma (DLCL) (e.g., diffuse large B-cell lymphoma (DLBCL)), follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma (CLL / SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphoma (e.g., mucosa-associated lymphoid tissue (MALT) lymphoma, nodal marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (i.e., "Waldenstrom's macroglobulinemia ( macroglobulinemia)”), hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma, and primary central nervous system (CNS) lymphoma; and T-cell NHL such as precursor T-lymphoblastic lymphoma / leukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g., mycosis fungiodes, Sezary syndrome), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy-type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplastic large cell lymphoma); mixtures of one or more leukemias / lymphomas as described above;and multiple myeloma (MM)), heavy chain disease (e.g., alpha chain disease, gamma chain disease, mu chain disease), hemangioblastoma, inflammatory myofibroblastic tumor, immune cell amyloidosis, kidney cancer (e.g., Wilms' tumor, renal cell carcinoma), liver cancer (e.g., hepatocellular carcinoma (HCC), malignant hepatoma), lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), lung adenocarcinoma), leiomyosarcoma (LMS), mastocytosis (e.g., systemic mastocytosis), myelodysplastic syndrome (MD S), mesothelioma, myeloproliferative disorders (MPDs) (e.g., polycythemia vera (PV), essential thrombocythemia (ET), idiopathic myeloid metaplasia (AMM) also known as myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic leukemia syndrome (HES)), neuroblastoma, neurofibromas (e.g., neurofibromatosis type 1 or type 2 (NF), schwannomatosis), neuroendocrine cancers (e.g., gastroenteropancreatic neuroendocrine cancers), ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma, ovarian clear cell carcinoma, ovarian serous cystadenocarcinoma), papillary adenocarcinoma, pancreatic cancer (e.g., pancreatic adenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), islet cell tumor), penile cancer (e.g., Paget's disease of the penis and scrotum), pineal tumor, primary neuroectodermal tumor (PNT), prostate cancer (e.g., prostate adenocarcinoma), rectal cancer, rhabdomyosarcoma, salivary duct carcinoma, skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)), small intestinal cancer (e.g., adnexal cancer), soft tissue sarcoma (e.g., malignant fibrous histiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous gland cancer, sweat gland cancer, synovioma, testicular cancer (e.g., seminoma, testicular embryonal carcinoma), thyroid cancer (e.g., papillary thyroid carcinoma, papillary thyroid carcinoma (PTC), medullary thyroid cancer), urethral cancer, vaginal cancer, and vulvar cancer (e.g., Paget's disease of the vulva), medulloblastoma, adenoid cystic carcinoma, melanoma, glioblastoma. ;

[0060] In some preferred embodiments, the present invention provides a compound represented by general formula (I), general formula (Ia), general formula (II) or general formula (III) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, or a pharmaceutical composition comprising the same for use in a method for treating a disease associated with CDK12 / 13 and in the preparation of a medicament for treating a disease associated with CDK12 / 13, wherein the disease associated with CDK12 / 13 includes but is not limited to: breast cancer, esophageal cancer, bladder cancer, lung cancer, hematopoietic system cancer, lymphoma, medulloblastoma, medulloblastoma, rectal adenocarcinoma, colon cancer, gastric cancer, pancreatic cancer, liver cancer, adenoid cystic carcinoma, prostate cancer, lung cancer, head and neck squamous cell carcinoma, brain cancer, hepatocellular carcinoma, melanoma, oligodendroglioma, glioblastoma, testicular cancer, ovarian clear cell carcinoma, ovarian serous cystadenocarcinoma, thyroid cancer, multiple myeloma (AML), renal cell carcinoma, mantle cell lymphoma, triple-negative breast cancer, and non-small cell lung cancer.

[0061] In some specific embodiments, the CDK12 / 13-mediated cancer is breast cancer, liver cancer, small cell lung cancer, non-small cell lung cancer, colon cancer, kidney cancer, bladder cancer, head and neck cancer, thyroid cancer, esophageal cancer, gastric cancer, pancreatic cancer, ovarian cancer, gallbladder cancer, cervical cancer, prostate, skin cancer, blood system tumors, nervous system tumors, melanoma, osteosarcoma or Kaposi's sarcoma.

[0062] Definition of terms

[0063] Unless stated otherwise, the terms used in the specification and claims have the following meanings.

[0064] The "hydrogen", "carbon" and "oxygen" in the compounds of the present invention include all isotopes thereof. Isotopes should be understood to include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include protium, tritium and deuterium, and isotopes of carbon include 12 C. 13 C and 14 C, oxygen isotopes include 16 O and 18 O etc.

[0065] The "isomer" of the present invention refers to a molecule with the same atomic composition and connection mode, but different three-dimensional spatial arrangement, including but not limited to diastereomers, enantiomers, cis-trans isomers, and mixtures thereof, such as racemic mixtures. Many organic compounds exist in optically active forms, that is, they have the ability to rotate the plane of plane polarized light. When describing optically active compounds, the prefix D, L or R, S is used to indicate the absolute configuration of the chiral center of the molecule. The prefix D, L or (+), (-) is used to name the sign of rotation of plane polarized light of the compound, (-) or L means that the compound is left-handed, and the prefix (+) or D means that the compound is right-handed. The chemical structure of these stereoisomers is the same, but their stereostructures are different. Specific stereoisomers can be enantiomers, and a mixture of isomers is usually called an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or a racemate, which may result in no stereoselectivity or stereospecificity during chemical reactions. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers, devoid of optical activity.

[0066] Depending on the choice of starting materials and process, the compounds of the invention may exist in the form of one of the possible isomers or a mixture thereof, such as a racemate and a diastereomeric mixture (depending on the number of asymmetric carbon atoms). Optically active (R)- or (S)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques.

[0067] Any resulting mixture of stereoisomers can be separated into the pure or substantially pure geometric isomers, enantiomers, diastereomers on the basis of the differences in the constituent physicochemical properties, for example, by chromatography and / or fractional crystallization.

[0068] The "halogen" of the present invention refers to fluorine, chlorine, bromine, and iodine. The "halogenated" of the present invention refers to substitution with fluorine, chlorine, bromine, or iodine.

[0069] The "alkyl" of the present invention refers to a straight or branched saturated aliphatic hydrocarbon group, preferably a straight or branched group containing 1 to 6 carbon atoms, and further preferably a straight or branched group containing 1 to 3 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, etc. The alkyl group may be substituted or unsubstituted, and when substituted, the substituent may be at any available point of attachment.

[0070] The "alkenyl" of the present invention refers to a straight or branched hydrocarbon chain radical consisting only of carbon atoms and hydrogen atoms, containing at least one carbon-carbon double bond, and having 2 to 12 carbon atoms. In some embodiments, the alkenyl group contains 2 to 8 carbon atoms. In other embodiments, the alkenyl group contains 2 to 4 carbon atoms. The alkenyl group is connected to the rest of the molecule by a single bond, such as vinyl, allyl, but-1-enyl, pent-1-enyl, pent-1,4-dienyl, etc. The alkenyl group is optionally substituted with one or more other substituents, such as alkyl, halogen, cyano, nitro, oxygen, sulfur, imino, haloalkyl, alkoxy, cycloalkyl, heterocyclic radical, heteroaryl, heteroaryl, etc.

[0071] The "alkynyl" of the present invention refers to a straight or branched hydrocarbon chain radical consisting only of carbon atoms and hydrogen atoms, containing at least one carbon-carbon triple bond, and having 2 to 12 carbon atoms. In some embodiments, the alkynyl group comprises 2 to 8 carbon atoms. In other embodiments, the alkynyl group comprises 2 to 4 carbon atoms. The alkynyl group is connected to the rest of the molecule by a single bond, such as ethynyl, propynyl, butynyl, pentynyl, hexynyl, etc. The alkynyl group is optionally substituted with one or more other substituents, such as alkyl, halogen, cyano, nitro, oxygen, sulfur, imino, haloalkyl, alkoxy, cycloalkyl, heterocyclic, heteroaryl, heteroaryl, etc.

[0072] The "carbonyl group" and "acyl group" of the present invention both refer to -C(O)-.

[0073] The "sulfonyl group" of the present invention refers to -S(O)2-.

[0074] The "sulfonamide group" of the present invention refers to -S(O)2NH-.

[0075] The "haloalkyl group" of the present invention refers to an alkyl group substituted with at least one halogen.

[0076] The "hydroxyalkyl group" of the present invention refers to an alkyl group substituted with at least one hydroxy group.

[0077] The "alkoxy" of the present invention refers to -O-alkyl. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, n-propoxy, isopropoxy, isobutoxy, sec-butoxy, etc. Alkoxy can be optionally substituted or unsubstituted, and when substituted, the substituent can be at any available point of attachment.

[0078] The "cycloalkyl" of the present invention refers to a cyclic saturated hydrocarbon group. Suitable cycloalkyl groups may be substituted or unsubstituted monocyclic, bicyclic or tricyclic saturated hydrocarbon groups having 3 to 12 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0079] The "heterocyclyl" of the present invention refers to a group of a 3- to 12-membered non-aromatic ring system having 1 to 4 ring heteroatoms, each of which is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon ("3-12 membered heterocyclyl"). In heterocyclyl groups containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as long as the valence permits. The heterocyclyl group can be either monocyclic ("monocyclic heterocyclyl") or a fused, bridged or spiral ring system (e.g., a bicyclic system (also known as a "bicyclic heterocyclyl")) and can be saturated or partially unsaturated. Suitable heterocyclyls include, but are not limited to, piperidinyl, azetidinyl, aziridine, tetrahydropyrrolyl, piperazinyl, dihydroquinazolinyl, oxacyclopropyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, etc. Each example of a heterocyclyl group can be optionally substituted or unsubstituted, and when substituted, the substituent can be at any usable point of attachment.

[0080] The "aryl" of the present invention refers to an aromatic system that may contain a monocyclic or fused polycyclic ring, preferably a monocyclic or fused bicyclic ring, containing 6 to 12 carbon atoms, preferably containing about 6 to about 10 carbon atoms. Suitable aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, fluorenyl, indanyl. Aryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents may be at any usable point of attachment.

[0081] The "heteroaryl" of the present invention refers to an aromatic group in which at least one carbon atom is replaced by a heteroatom, preferably composed of 5-12 atoms (5-12-membered heteroaryl), and more preferably composed of 5-10 atoms (5-10-membered heteroaryl), wherein the heteroatom is O, S, or N. The heteroaryl groups include, but are not limited to, imidazolyl, pyrrolyl, furanyl, thienyl, pyrazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, tetrazolyl, indolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazinyl, isoindolyl, benzopyrazolyl, benzimidazolyl, benzofuranyl, benzopyranyl, benzothienyl, benzoxazolyl, benzothiazolyl, benzisoxazolyl, benzisothiazolyl, quinolyl, isoquinolyl, quinazolinyl, cinnolinyl, quinoxalinyl, benzoxazinyl, benzothiazinyl, imidazopyridinyl, pyrimidopyrazolyl, pyrimidoimidazolyl, etc. The heteroaryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents may be at any available point of attachment.

[0082] The "pharmaceutically acceptable salt" of the present invention refers to the salt of the compound of the present invention, which is safe and effective when used in mammals and has the desired biological activity.

[0083] The "solvate" of the present invention refers to a complex formed by the combination of a solute (such as an active compound, a salt of an active compound) and a solvent (such as water) in a conventional sense. The solvent refers to a solvent known to or easily determined by a person skilled in the art. If it is water, the solvate is usually referred to as a hydrate, such as a hemihydrate, a monohydrate, a dihydrate, a trihydrate or an alternative thereof.

[0084] The in vivo effects of compounds of formula (I), (Ia), (II) or (III) may be partially exerted by one or more metabolites formed in the human or animal body after administration of the compounds of formula (I), (Ia), (II) or (III). As described above, the in vivo effects of compounds of formula (I), (Ia), (II) or (III) may also be exerted via metabolism of precursor compounds ("prodrugs"). "Prodrugs" of the present invention refer to compounds that are converted into compounds of the present invention under physiological conditions in an organism by reaction with enzymes, gastric acid, etc., i.e., compounds that are converted into compounds of the present invention by oxidation, reduction, hydrolysis, etc. of enzymes and / or compounds that are converted into compounds of the present invention by hydrolysis reactions of gastric acid, etc.

[0085] The term "crystalline" in the present invention refers to a solid whose internal structure is formed by regularly repeating constituent atoms (or groups thereof) in three dimensions, and is distinguished from an amorphous solid that does not have such a regular internal structure.

[0086] The "pharmaceutical composition" of the present invention refers to a mixture comprising any one of the compounds of the present invention, including corresponding isomers, prodrugs, solvates, pharmaceutically acceptable salts or chemically protected forms thereof, and one or more pharmaceutically acceptable carriers and / or one or more other drugs. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to an organism. The composition is generally used to prepare a drug for the treatment and / or prevention of a disease mediated by one or more kinases.

[0087] The "pharmaceutically acceptable carrier" of the present invention refers to a carrier that does not cause significant irritation to an organism and does not interfere with the biological activity and properties of the administered compound, including all solvents, diluents or other excipients, dispersants, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, etc. Unless any conventional carrier medium is incompatible with the compound of the present invention. Some examples of pharmaceutically acceptable carriers include, but are not limited to, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, as well as cellulose and cellulose acetate; malt, gelatin, etc.

[0088] The "excipient" of the present invention refers to an inert substance added to a pharmaceutical composition to further facilitate administration of a compound. Excipients may include calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oils, polyethylene glycols. DETAILED DESCRIPTION

[0089] The present invention is further described in detail below in conjunction with the examples, but the present invention is not limited to these examples. The materials used in the following examples are commercially available unless otherwise specified.

[0090] Intermediate 1: 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)propanoic acid

[0091]

[0092] Step 1: Preparation of methyl 2-(4-bromo-1H-pyrazol-1-yl)propanoate

[0093]

[0094] Dissolve 4-bromo-1H-pyrazole (5.0 g, 34.02 mmol, 1.0 eq) in acetonitrile (50 mL), add potassium carbonate (9.4 g, 68.04 mmol, 2.0 eq), add methyl 2-bromopropionate (6.82 g, 40.82 mmol, 1.2 eq) dropwise under stirring, and leave overnight at room temperature. LC-MS tracking and monitoring the reaction, after the reaction is completed, add water to quench, add ethyl acetate to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate, and purify by column chromatography to obtain 6.4 g of the title compound. ESI-MS m / z: 232.9 [M+H] + .

[0095] Step 2: Preparation of methyl 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)propanoate

[0096]

[0097] Dissolve 1-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridin-1(2H)-yl)prop-2-en-1-one (1.4 g, 4.53 mmol, 1.0 eq) in dioxane (30 mL), add methyl 2-(4-bromo-1H-pyrazol-1-yl)propanoate (1.26 g, 5.43 mmol, 1.2 eq), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (0.37 g, 0.45 mmol, 0.1 eq), cesium carbonate (4.4 g, 13.5 mmol, 3 eq) and water (3 mL) in sequence, and stir at 90 °C for 3 h under argon protection. The reaction was monitored by LC-MS. After the reaction was completed, water was added to quench the reaction, ethyl acetate was added to extract, the organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 900 mg of the title compound. ESI-MS m / z: 290.0 [M+H] + .

[0098] Step 3: Preparation of 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)propanoic acid

[0099]

[0100] Dissolve methyl 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)propanoate (0.9 g, 3.11 mmol, 1.0 eq) in tetrahydrofuran (8 mL), add water (4 mL) and lithium hydroxide (0.3 g, 12.5 mmol, 4.0 eq), and stir at room temperature for 4 h. LC-MS tracking and monitoring the reaction, after the reaction is completed, add water to quench, adjust the pH to 5 with dilute hydrochloric acid, add ethyl acetate to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate to obtain 0.6 g of the title compound. ESI-MS m / z: 276.0 [M+H] + .

[0101] Intermediate 2: 2-(2-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)thiazol-4-yl)propanoic acid

[0102]

[0103] Step 1: Preparation of methyl 2-(4-bromo-1H-pyrazol-1-yl)propanoate

[0104]

[0105] Add 2-amino-4-thiazoleacetic acid methyl ester (96.7g, 0.56mol, 1.0eq) and cuprous bromide (241.0g, 1.68mol, 3.0eq) into a three-necked flask, protect with argon, add acetonitrile (1.2L), cool to 0°C, slowly add tert-butyl nitrite (173.7g, 1.68mol, 3.0eq), stir for 30min, and stir at room temperature for 3h. LC-MS tracking and monitoring the reaction, after the reaction is completed, add water to quench, add ethyl acetate to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate, and purify by column chromatography to obtain 75.1g of the title compound. ESI-MS m / z: 236.0[M+H] + .

[0106] Step 2: Preparation of methyl 2-(2-bromothiazol-4-yl)propanoate

[0107]

[0108] Dissolve methyl 2-(4-bromo-1H-pyrazol-1-yl)propanoate (75.1 g, 0.32 mol, 1.0 eq) in anhydrous tetrahydrofuran (800 mL), protect with argon, cool to -78 ° C, slowly add 2M lithium diisopropylamide (240 mL, 0.48 mol, 1.5 eq) and stir for 1 h, add iodomethane (36.3 g, 0.26 mol, 0.8 eq) dropwise, stir at room temperature for 2 h. LC-MS monitors the reaction, and after the reaction is complete, add saturated ammonium chloride solution to quench the reaction, add ethyl acetate to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate, and purify by column chromatography to obtain 27.1 g of the title compound. ESI-MS m / z: 249.9 [M+H] + .

[0109] Step 3: Preparation of methyl 2-(2-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)thiazol-4-yl)propanoate

[0110]

[0111] Dissolve 1-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridin-1(2H)-yl)prop-2-en-1-one (2.3 g, 8.84 mmol, 1.2 eq) in dioxane (30 mL), add methyl 2-(2-bromothiazol-4-yl)propanoate (2.0 g, 8.03 mmol, 1.0 eq), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (0.29 g, 0.80 mmol, 0.1 eq), cesium carbonate (7.8 g, 124.1 mmol, 3 eq) and water (3 mL) in sequence, and stir at 90 °C for 3 h under argon protection. The reaction was monitored by LC-MS. After the reaction was completed, water was added to quench the reaction, ethyl acetate was added to extract, the organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 1.5 g of the title compound. ESI-MS m / z: 306.9 [M+H] + .

[0112] Step 4: Preparation of 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)propanoic acid

[0113]

[0114] Dissolve methyl 2-(2-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)thiazol-4-yl)propanoate (1.5 g, 4.90 mmol, 1.0 eq) in tetrahydrofuran (30 mL), add water (10 mL) and lithium hydroxide (0.47 g, 19.6 mmol, 4.0 eq), and stir at room temperature for 4 h. LC-MS tracking and monitoring the reaction, after the reaction is completed, add water to quench, adjust the pH to 5 with dilute hydrochloric acid, add ethyl acetate to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate to obtain 0.8 g of the title compound. ESI-MS m / z: 293.1 [M+H] + .

[0115] Example 1: 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)-N-(4-(5,6-difluoro-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide

[0116]

[0117] Step 1: Preparation of tert-butyl 5,6-difluoro-1H-indole-1-carboxylate

[0118]

[0119] Dissolve 5,6-difluoro-1H-indole (2.0 g, 0.013 mol, 1.0 eq) in anhydrous dichloromethane (20 mL), add 4-dimethylaminopyridine (320 mg, 2.6 mmol, 0.2 eq), triethylamine (1.3 g, 0.013 mol, 1.0 eq), di-tert-butyl carbonic anhydride (4.3 g, 0.02 mol, 1.5 eq) in dichloromethane (10 mL) in turn, and react at room temperature for 4 h. LC-MS tracking and monitoring the reaction, after the reaction is completed, add water to quench, add dichloromethane to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate, and purify by column chromatography to obtain 3.3 g of the title compound. ESI-MS m / z: 154.2 [M+H-Boc] + .

[0120] Step 2: Preparation of tert-butyl 3-bromo-5,6-difluoro-1H-indole-1-carboxylate

[0121]

[0122] Dissolve tert-butyl 5,6-difluoro-1H-indole-1-carboxylate (3.3 g, 0.013 mol, 1.0 eq) in anhydrous N,N-dimethylformamide (10 mL), add N-bromosuccinimide (2.44 g, 0.014 mol, 1.05 eq) in batches, and react at room temperature overnight. LC-MS tracking and monitoring the reaction, after the reaction is completed, add water to quench, add ethyl acetate to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate, and purify by column chromatography to obtain 2.3 g of the title compound. ESI-MS m / z: 332.1 [M+H] + .

[0123] Step 3: Preparation of tert-butyl 5,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole-1-carboxylate

[0124]

[0125] Dissolve tert-butyl 3-bromo-5,6-difluoro-1H-indole-1-carboxylate (2.3g, 6.9mmol, 1.0eq) in dioxane (20mL), add [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (512mg, 0.7mmol, 0.1eq), potassium acetate (1.7g, 17.4mmol, 2.5eq) and biboric acid pinacol ester (3.5g, 14mmol, 2.0eq) in sequence, and stir at 80°C for 3h under argon protection. LC-MS tracking and monitoring the reaction, after the reaction is completed, add water to quench, add ethyl acetate to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate, and purify by column chromatography to obtain 0.6g of the title compound. ESI-MS m / z: 380.2[M+H] + .

[0126] Step 4: Preparation of tert-butyl 3-(2-amino-5-(trifluoromethyl)pyrimidin-4-yl)-5,6-difluoro-1H-indole-1-carboxylate

[0127]

[0128] 4-Chloro-5-(trifluoromethyl)pyrimidin-2-amine (0.16 g, 0.881 mmol, 1.1 eq) was dissolved in dioxane (20 mL), and 5,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester (280 mg, 0.74 mmol, 1.0 eq) was added in sequence.

[0129] [1,1'-Bis(diphenylphosphino)ferrocene]palladium dichloride (60mg, 0.074mmol, 0.1eq), cesium carbonate (721.5mg, 2.22mmol, 3.0eq) and water (2mL) were stirred at 90°C for 3h under argon protection. The reaction was monitored by LC-MS tracking. After the reaction was completed, water was added to quench the reaction, and ethyl acetate was added to extract the reaction. The organic phases were combined, washed with water three times, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 300mg of the title compound. ESI-MS m / z: 414.9[M+H] + .

[0130] Step 5: Preparation of tert-butyl 3-(2-(2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)propionamido)-5-(trifluoromethyl)pyrimidin-4-yl)-5,6-difluoro-1H-indole-1-carboxylate

[0131]

[0132] Dissolve tert-butyl 3-(2-amino-5-(trifluoromethyl)pyrimidin-4-yl)-5,6-difluoro-1H-indole-1-carboxylate (0.1g, 0.24mmol, 0.6eq) in anhydrous dichloromethane (10mL), add intermediate 1 (0.12g, 0.41mmol, 1.0eq) and pyridine (10mL), cool to -20°C, add phosphorus oxychloride (74mg, 0.48mmol, 1.2eq) dropwise and stir for 2h. LC-MS tracking and monitoring the reaction, after the reaction is completed, the reaction solution is poured into ice water, extracted with dichloromethane, the organic phases are combined, the organic phases are washed three times with water, dried and concentrated over anhydrous sodium sulfate, and purified by column chromatography to obtain 38mg of the title compound. ESI-MS m / z: 672.2[M+H] + .

[0133] Step 6: Preparation of 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)-N-(4-(5,6-difluoro-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide

[0134]

[0135] Dissolve tert-butyl 3-(2-(2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)propionamido)-5-(trifluoromethyl)pyrimidin-4-yl)-5,6-difluoro-1H-indole-1-carboxylate (38 mg, 0.057 mmol, 1.0 eq) in dichloromethane (2 mL), cool to 0°C in an ice bath, add trifluoroacetic acid (456 mg, 4 mmol) dropwise, and react at room temperature for 1 h. LC-MS was used to monitor the reaction. After the reaction was completed, the reaction solution was dried and purified by C18 column chromatography to obtain 20 mg of the title compound. ESI-MS m / z: 571.9 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ12.09(s,1H),11.34(s,1H),9.06-8.91(m,2H),8.20-8.04(m,2H),7.77-7.66(m,1H),7.58-7.45(m,1H),7.04-6.82(m,1 H),6.24-6.09(m,2H),5.77-5.66(m,1H),5.56-5.45(m,1H),4.31(d,J=1 6.0Hz,2H),3.74-3.60(m,2H),2.30-2.18(m,2H),1.75(d,J=8.0Hz,3H).

[0136] Example 2: N-(4-(1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)-2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)propanamide

[0137]

[0138] The preparation method is the same as that of Example 1, except that the raw material 5,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester is replaced with 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester to obtain the title compound. ESI-MS m / z: 536.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ12.00(s,1H),11.20(d,J=12.0Hz,1H),8.94(s,1H),8.78(d,J=8 .0Hz,1H),8.17-8.04(m,1H),7.99(s,1H),7.77-7.65(m,1H),7.50(d,J=8.0Hz,1H),7.2 7-7.13(m,2H),7.04-6.81(m,1H),6.23-6.09(m,2H),5.76-5.66(m,1H),5.58-5.48(m,1 H),4.31(d,J=16.0Hz,2H),3.73-3.61(m,2H),2.30-2.17(m,2H),1.75(d,J=4.0Hz,3H).

[0139] The racemate (200 mg) of N-(4-(1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)-2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)propanamide was separated by chiral preparative column. (Method: Chiral column: ChiralPak AD, 250×30 mm ID, 10 μm, elution: isocratic: B 40%, A=carbon dioxide, B=isopropanol) to obtain Isomer 1 (53 mg) and Isomer 2 (82 mg) of the title compound.

[0140] Isomer 1 (Example 3): ESI-MS m / z: 536.0 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ12.00(s,1H),11.20(d,J=12.0Hz,1H),8.94(s,1H),8.78(d,J=8.0Hz,1H),8.17-8.04(m,1H) ,7.99(s,1H),7.77-7.65(m,1H),7.50(d,J=8.0Hz,1H),7.27-7.13(m,2H),7.04-6.81(m,1H),6.23-6.09(m,2H),5.76 -5.66(m,1H),5.58-5.48(m,1H),4.31(d,J=16.0Hz,2H),3.73-3.61(m,2H),2.30-2.17(m,2H),1.75(d,J=4.0Hz,3H).

[0141] Isomer 2 (Example 4): ESI-MS m / z: 536.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ12.00(s,1H),11.20(d,J=12.0Hz,1H),8.94(s,1H),8.78(d,J=8.0Hz,1H),8.17-8.04(m,1H) ,7.99(s,1H),7.77-7.65(m,1H),7.50(d,J=8.0Hz,1H),7.27-7.13(m,2H),7.04-6.81(m,1H),6.23-6.09(m,2H),5.76 -5.66(m,1H),5.58-5.48(m,1H),4.31(d,J=16.0Hz,2H),3.73-3.61(m,2H),2.30-2.17(m,2H),1.75(d,J=4.0Hz,3H).

[0142] Example 5: 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)-N-(5-chloro-4-methoxypyrimidin-2-yl)propanamide

[0143]

[0144] The preparation method is the same as that of Example 1, except that the raw material 3-(2-amino-5-(trifluoromethyl)pyrimidin-4-yl)-5,6-difluoro-1H-indole-1-carboxylic acid tert-butyl ester is replaced with 5-chloro-4-methoxypyrimidin-2-amine to obtain the title compound. ESI-MS m / z: 416.9 [M+H] + 。 1HNMR(400MHz, DMSO-d6)δ11.00(s,1H),8.51(s,1H),8.05(d,J=28.0Hz,1H),7.68(d,J=16.0Hz,1H),7.00-6.84(m,1H),6.20-6.10(m,2H),5 .74-5.67(m,1H),5.45-5.32(m,1H),4.29(d,J=16.0Hz,2H),4.00(s,3H),3.71-3.60(m,2H),2.23(d,J=16.0Hz,2H),1.70(d,J=8.0Hz,3H).

[0145] Example 6: 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)-N-(5-chloro-4-(1H-indol-3-yl)pyrimidin-2-yl)propanamide

[0146]

[0147] The preparation method is the same as that of Example 1, except that the raw material 3-(2-amino-5-(trifluoromethyl)pyrimidin-4-yl)-5,6-difluoro-1H-indole-1-carboxylic acid tert-butyl ester is replaced with 3-(2-amino-5-chloropyrimidin-4-yl)-1H-indole-1-carboxylic acid tert-butyl ester to obtain the title compound. ESI-MS m / z: 501.9 [M+H] + 。 1 H NMR (400MHz, DMSO-d6) δ12.03(s,1H),10.94(d,J=12.0Hz,1H),9.06(d,J=8.0Hz,1H) ,8.65(s,2H),8.20-8.05(m,1H),7.78-7.67(m,1H),7.50(d,J=4.0Hz,1H),7.28-7.1 4(m,2H),7.06-6.82(m,1H),6.24-6.10(m,2H),5.77-5.66(m,1H),5.55-5.43(m,1H) ,4.32(d,J=16.0Hz,2H),3.73-3.62(m,2H),2.31-2.16(m,2H),1.76(d,J=8.0Hz,3H).

[0148] Example 7: N-(4-(1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)-2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl]-1H-pyrazol-1-yl)propanamide

[0149]

[0150] The preparation method is the same as that of Example 1, except that the raw material 5,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester is replaced with 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-pyrrolo[2,3-b]pyridine-1-carboxylic acid tert-butyl ester to obtain the title compound. ESI-MS m / z: 536.9 [M+H] + 。 1 H NMR (400MHz, DMSO-d6) δ12.59 (s, 1H), 11.30 (s, 1H), 9.26 (d, J = 8.0Hz, 1H), 8.98 (s, 1H),8.37-8.31(m,1H),8.08(s,1H),7.76-7.67(m,1H),7.29-7.21(m,1H),7.04-6. 88(m,1H),6.20-6.10(m,2H),5.75-5.67(m,1H),5.55-5.47(m,1H),5.37-5.30(m,1 H),4.38-4.26(m,2H),3.73-3.61(m,2H),2.04-1.95(m,2H),1.76(d,J=8.0Hz,3H).

[0151] Example 8: 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)-N-(4-(7-methoxy-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide

[0152]

[0153] Step 1: Preparation of 3-iodo-7-methoxy-1H-indole

[0154]

[0155] Dissolve 7-methoxy-1H-indole (1.0 g, 6.8 mmol, 1.0 eq) in N,N-dimethylformamide (6 mL), add potassium hydroxide (0.95 g, 17.0 mmol, 2.5 eq), then dropwise add iodine (1.74 g, 6.9 mmol, 1.01 eq) in N,N-dimethylformamide (6 mL), and react at room temperature for 1 h. LC-MS tracking and monitoring the reaction, after the reaction is completed, add water to quench, add ethyl acetate to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate, and purify by column chromatography to obtain 1.8 g of the title compound. ESI-MS m / z: 273.9 [M+H] + .

[0156] Step 2: Preparation of tert-butyl 3-iodo-7-methoxy-1H-indole-1-carboxylate

[0157]

[0158] Dissolve 3-iodo-7-methoxy-1H-indole (1.8 g, 6.6 mmol, 1.0 eq) in anhydrous dichloromethane (10 mL), add 4-dimethylaminopyridine (80.56 mg, 0.7 mmol, 0.1 eq) and di-tert-butyl carbonic anhydride (2.16 g, 9.9 mmol, 2.5 eq) in dichloromethane (10 mL) in turn, and react the solution at room temperature for 0.5 h. LC-MS tracking and monitoring the reaction, after the reaction is completed, add water to quench, add ethyl acetate to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate, and purify by column chromatography to obtain 2.2 g of the title compound. ESI-MS m / z: 317.9 [M+H-Boc] + .

[0159] Step 3: Preparation of tert-butyl 7-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole-1-carboxylate

[0160]

[0161] Dissolve tert-butyl 3-iodo-7-methoxy-1H-indole-1-carboxylate (2.0 g, 5.36 mmol, 1.0 eq) in dioxane (20 mL), add tetrakistriphenylphosphine palladium (187 mg, 0.16 mmol, 0.03 eq), triethylamine (5.5 g, 53.6 mmol, 10.0 eq) and 4,4,5,5-tetramethyl-1,3,2-dioxaborane (1.03 g, 8.04 mmol, 1.5 eq) in turn, and stir at 80 ° C for 6 h under argon protection. LC-MS tracking and monitoring the reaction, after the reaction is completed, add water to quench, add ethyl acetate to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate, and purify by column chromatography to obtain 1.0 g of the title compound. ESI-MS m / z: 318.2 [M+H-Boc] + .

[0162] Step 4: Preparation of 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)-N-(4-(7-methoxy-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide

[0163]

[0164] The preparation method is the same as that of Example 1, except that the raw material 5,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester is replaced with 7-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester to obtain the title compound. ESI-MS m / z: 565.9 [M+H] + 。 1 H NMR (400MHz, DMSO-d6) δ12.20(s,1H),11.20(s,1H),8.94(s,1H),8.33(d,J=8.0Hz ,1H),8.16-8.04(m,1H),7.81(s,1H),7.75-7.65(m,1H),7.15-7.05(m,1H),7.03-6 .77(m,2H),6.23-6.10(m,2H),5.76-5.67(m,1H),5.58-5.49(m,1H),4.31(d,J=16. 0Hz,2H),3.95(s,3H),3.72-3.62(m,2H),2.30-2.18(m,2H),1.75(d,J=4.0Hz,3H).

[0165] The racemate (150 mg) of 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)-N-(4-(7-methoxy-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide was separated by chiral preparative column. (Method: Chiral column: ChiralPak AD, 250×30 mm ID, 10 μm, elution: isocratic: B 40%, A=carbon dioxide, B=isopropanol) to obtain Isomer 1 (67 mg) and Isomer 2 (68 mg) of the title compound.

[0166] Isomer 1 (Example 9): ESI-MS m / z: 565.9 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ12.20(s,1H),11.20(s,1H),8.94(s,1H),8.33(d,J=8.0Hz ,1H),8.16-8.04(m,1H),7.81(s,1H),7.75-7.65(m,1H),7.15-7.05(m,1H),7.03-6 .77(m,2H),6.23-6.10(m,2H),5.76-5.67(m,1H),5.58-5.49(m,1H),4.31(d,J=16. 0Hz,2H),3.95(s,3H),3.72-3.62(m,2H),2.30-2.18(m,2H),1.75(d,J=4.0Hz,3H).

[0167] Isomer 2 (Example 10): ESI-MS m / z: 565.9 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ12.20(s,1H),11.20(s,1H),8.94(s,1H),8.33(d,J=8.0Hz ,1H),8.16-8.04(m,1H),7.81(s,1H),7.75-7.65(m,1H),7.15-7.05(m,1H),7.03-6 .77(m,2H),6.23-6.10(m,2H),5.76-5.67(m,1H),5.58-5.49(m,1H),4.31(d,J=16. 0Hz,2H),3.95(s,3H),3.72-3.62(m,2H),2.30-2.18(m,2H),1.75(d,J=4.0Hz,3H).

[0168] Example 11: 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)-N-(4-(7-methyl-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide

[0169]

[0170] The preparation method is the same as that of Example 1, except that the raw material 5,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester is replaced with 7-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester to obtain the title compound. ESI-MS m / z: 549.9 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ12.02(s,1H),11.20(d,J=12.0Hz,1H),8.95(s,1H),8.57( d,J=8.0Hz,1H),8.15-8.03(m,1H),7.91(s,1H),7.70(d,J=16.0Hz,1H),7.11-6.8 3(m,3H),6.23-6.07(m,2H),5.76-5.66(m,1H),5.58-5.48(m,1H),4.30(d,J=16.0 Hz,2H),3.73-3.61(m,2H),2.51(s,3H),2.29-2.17(m,2H),1.75(d,J=8.0Hz,3H).

[0171] The racemate of 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)-N-(4-(7-methyl-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide (200 mg) was separated by chiral preparative column. (Method: Chiral column: ChiralCel OD, 250×30 mm ID, 10 μm, elution: isocratic: B 40%, A=carbon dioxide, B=ethanol) to obtain Isomer 1 (77 mg) and Isomer 2 (76 mg) of the title compound.

[0172] Isomer 1 (Example 12): ESI-MS m / z: 549.9 [M+H] + . 1H NMR(400MHz, DMSO-d6)δ12.02(s,1H),11.20(d,J=12.0Hz,1H),8.95(s,1H),8.57( d,J=8.0Hz,1H),8.15-8.03(m,1H),7.91(s,1H),7.70(d,J=16.0Hz,1H),7.11-6.8 3(m,3H),6.23-6.07(m,2H),5.76-5.66(m,1H),5.58-5.48(m,1H),4.30(d,J=16.0 Hz,2H),3.73-3.61(m,2H),2.51(s,3H),2.29-2.17(m,2H),1.75(d,J=8.0Hz,3H).

[0173] Isomer 2 (Example 13): ESI-MS m / z: 549.9 [M+H] + 。 1 H NMR(400MHz, DMSO-d6)δ12.02(s,1H),11.20(d,J=12.0Hz,1H),8.95(s,1H),8.57( d,J=8.0Hz,1H),8.15-8.03(m,1H),7.91(s,1H),7.70(d,J=16.0Hz,1H),7.11-6.8 3(m,3H),6.23-6.07(m,2H),5.76-5.66(m,1H),5.58-5.48(m,1H),4.30(d,J=16.0 Hz,2H),3.73-3.61(m,2H),2.51(s,3H),2.29-2.17(m,2H),1.75(d,J=8.0Hz,3H).

[0174] Example 14: 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)-N-(4-(6-cyano-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide

[0175]

[0176] The preparation method is the same as that of Example 1, except that the raw material 5,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester is replaced with 6-cyano-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester to obtain the title compound. ESI-MS m / z: 560.9 [M+H] +。 1 H NMR(400MHz,DMSO-d6)δ12.43(s,1H),11.35(s,1H),9.06-8.88(m,2H),8.2 3(s,1H),8.16-8.00(m,2H),7.76-7.65(m,1H),7.54-7.45(m,1H),7.03-6.8 3(m,1H),6.25-6.10(m,2H),5.78-5.66(m,1H),5.58-5.46(m,1H),4.31(d, J=16.0Hz,2H),3.74-3.62(m,2H),2.30-2.18(m,2H),1.75(d,J=4.0Hz,3H).

[0177] Example 15: N-(4-(1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)-2-(4-(1-acryloyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazol-1-yl)propanamide

[0178]

[0179] Step 1: Preparation of 2-(4-(1-acryloyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazol-1-yl)propanoic acid

[0180]

[0181] The preparation method is the same as that of intermediate 1, except that the raw material 1-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridin-1(2H)-yl)prop-2-en-1-one is replaced with 1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridin-1(2H)-yl)prop-2-en-1-one to obtain the title compound. ESI-MS m / z: 276.0[M+1] + 。

[0182] Step 2: Preparation of tert-butyl 3-(2-amino-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-1-carboxylate

[0183]

[0184] Dissolve 4-chloro-5-(trifluoromethyl)pyrimidin-2-amine (3.0 g, 15.2 mmol, 1.0 eq) in dioxane (160 mL), add tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole-1-carboxylate (5.5 g, 15.99 mmol, 1.1 eq), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (0.56 g, 0.076 mmol, 0.05 eq), cesium carbonate (14.8 g, 45.6 mmol, 3.0 eq) and water (16 mL) in sequence, stir at 100 ° C for 3 h under argon protection. LC-MS tracking and monitoring the reaction, after the reaction is completed, add water to quench, add ethyl acetate to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate, and purify by column chromatography to obtain 3.5 g of the title compound. ESI-MS m / z:379.1[M+1] + .

[0185] Step 3: Preparation of tert-butyl 3-(2-(2-(4-(1-acryloyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazol-1-yl)propionamido)-5-(trifluoromethyl)pyrimidin-4-yl)-1-indole-1-carboxylate

[0186]

[0187] 2-(4-(1-acryloyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazol-1-yl)propionic acid (275 mg, 1.0 mmol, 1.0 eq) was dissolved in anhydrous acetonitrile (10 mL), tetramethyl chlorourea hexafluorophosphate (504 mg, 1.8 mmol, 1.8 eq) and N-methylimidazole (287 mg, 3.5 mmol, 3.5 eq) were added in sequence, and stirred at room temperature for 2 h, and then tert-butyl 3-(2-amino-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-1-carboxylate (227 mg, 0.6 mmol, 0.6 eq) was added, and stirred at room temperature for 3 h. LC-MS was used to monitor the reaction. After the reaction was completed, water was added to quench the reaction, and ethyl acetate was added for extraction. The organic phases were combined, washed with water three times, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 130 mg of the title compound. ESI-MS m / z:636.1[M+H] + .

[0188] Step 4: Preparation of N-(4-(1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)-2-(4-(1-acryloyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazol-1-yl)propanamide

[0189]

[0190] Dissolve tert-butyl 3-(2-(2-(4-(1-acryloyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazol-1-yl)propionamido)-5-(trifluoromethyl)pyrimidin-4-yl)-1-indole-1-carboxylate (130 mg, 0.2 mmol, 1.0 eq) in dichloromethane (10 mL), cool to 0°C in an ice bath, add trifluoroacetic acid (8 mL) dropwise, and react at room temperature for 1 h. LC-MS was used to monitor the reaction. After the reaction was completed, the reaction solution was dried and purified by C18 column chromatography to obtain 34 mg of the title compound. ESI-MS m / z: 536.0 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ12.00(s,1H),11.18(s,1H),8.94(s,1H),8.77(d,J=8.0Hz,1H) ,8.05-7.95(m,2H),7.69-7.63(m,1H),7.50(d,J=8.0Hz,1H),7.26-7.13(m,2H),6.93-6 .74(m,1H),6.17-6.08(m,1H),6.05-5.97(m,1H),5.69(dd,J=12.0,4.0Hz,1H),5.52(q, J=8.0Hz,1H),4.25-4.07(m,2H),3.77-3.67(m,2H),2.07(s,2H),1.73(d,J=4.0Hz,3H).

[0191] Example 16: 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)-N-(4-(6-trifluoromethyl-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide

[0192]

[0193] The preparation method is the same as that of Example 1, except that the raw material 5,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester is replaced with 6-trifluoromethyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester to obtain the title compound. ESI-MS m / z: 603.9 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ12.32 (s, 1H), 11.30 (d, J = 16.0Hz, 1H), 9.05-8.92 (m, 2 H),8.25-8.05(m,2H),7.86(s,1H),7.75-7.66(m,1H),7.43(d,J=8.0Hz,1H),7. 03-6.83(m,1H),6.19-6.12(m,2H),5.70(d,J=12.0Hz,1H),5.55-5.46(m,1H), 4.36-4.24(m,2H),3.72-3.60(m,2H),2.28-2.18(m,2H),1.76(d,J=8.0Hz,3H).

[0194] The racemate of 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)-N-(4-(6-trifluoromethyl-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide (200 mg) was separated by chiral preparative column. (Method: Chiral column: ChiralCel OD, 250×30 mm ID, 10 μm, elution: isocratic: B 40%, A=carbon dioxide, B=ethanol (0.1% ammonia water)) to obtain isomer 1 (52 mg) and isomer 2 (60 mg) of the title compound.

[0195] Isomer 1 (Example 17): ESI-MS m / z: 603.9 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ12.32 (s, 1H), 11.30 (d, J = 16.0Hz, 1H), 9.05-8.92 (m, 2 H),8.25-8.05(m,2H),7.86(s,1H),7.75-7.66(m,1H),7.43(d,J=8.0Hz,1H),7. 03-6.83(m,1H),6.19-6.12(m,2H),5.70(d,J=12.0Hz,1H),5.55-5.46(m,1H), 4.36-4.24(m,2H),3.72-3.60(m,2H),2.28-2.18(m,2H),1.76(d,J=8.0Hz,3H).

[0196] Isomer 2 (Example 18): ESI-MS m / z: 603.9 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ12.32 (s, 1H), 11.30 (d, J = 16.0Hz, 1H), 9.05-8.92 (m, 2 H),8.25-8.05(m,2H),7.86(s,1H),7.75-7.66(m,1H),7.43(d,J=8.0Hz,1H),7. 03-6.83(m,1H),6.19-6.12(m,2H),5.70(d,J=12.0Hz,1H),5.55-5.46(m,1H), 4.36-4.24(m,2H),3.72-3.60(m,2H),2.28-2.18(m,2H),1.76(d,J=8.0Hz,3H).

[0197] Example 19: 2-(4-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-1H-pyrazol-1-yl)-N-(4-(5-trifluoromethyl-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide

[0198]

[0199] The preparation method is the same as that of Example 1, except that the raw material 5,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester is replaced with 5-trifluoromethyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester to obtain the title compound. ESI-MS m / z: 603.9 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ12.33(s,1H),11.18(d,J=12.0Hz,1H),9.24(s,1H),8.99 (s,1H),8.16(s,1H),8.11-7.99(m,1H),7.78-7.63(m,2H),7.57-7.48(m,1H),6. 99-6.81(m,1H),6.22-6.06(m,2H),5.70(d,J=12.0Hz,1H),5.53-5.43(m,1H),4. 37-4.21(m,2H),3.73-3.59(m,2H),2.23(d,J=20.0Hz,2H),1.73(d,J=4.0Hz,3H).

[0200] Example 20: 2-(2-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)thiazol-4-yl)-N-(4-(7-methoxy-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide

[0201]

[0202] Step 1: Preparation of tert-butyl 3-(2-(2-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-thiazol-4-yl)propionamido)-5-(trifluoromethyl)pyrimidin-4-yl)-7-methoxy-1H-indole-1-carboxylate

[0203]

[0204] Intermediate 2 (336 mg, 1.15 mmol, 1.0 eq) was dissolved in 1,2-dichloroethane (20 mL) and added to a microwave tube. Tetramethyl chlorourea hexafluorophosphate (386 mg, 1.38 mmol, 1.2 eq), N-methylimidazole (445 mg, 3.45 mmol, 3.0 eq) and tert-butyl 3-(2-amino-5-(trifluoromethyl)pyrimidin-4-yl)-7-methoxy-1H-indole-1-carboxylate (564 mg, 1.38 mmol, 1.2 eq) were added in sequence and stirred at 85°C for 3 h. LC-MS was used to monitor the reaction. After the reaction was completed, water was added to quench the reaction, and ethyl acetate was added to extract the reaction. The organic phases were combined, washed with water three times, dried over anhydrous sodium sulfate, and concentrated. The title compound (10 mg) was purified by column chromatography. ESI-MS m / z: 683.3 [M+H] + .

[0205] Step 2: 2-(2-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)thiazol-4-yl)-N-(4-(7-methoxy-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide

[0206]

[0207] Dissolve tert-butyl 3-(2-(2-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)-thiazol-4-yl)propionamido)-5-(trifluoromethyl)pyrimidin-4-yl)-7-methoxy-1H-indole-1-carboxylate (10 mg, 0.015 mmol, 1.0 eq) in dichloromethane (1 mL), cool to 0°C in an ice bath, add trifluoroacetic acid (0.5 mL) dropwise, and react at room temperature for 1 h. LC-MS was used to monitor the reaction. After the reaction was completed, the reaction solution was dried and purified by C18 column chromatography to obtain 2 mg of the title compound. ESI-MS m / z: 583.2[M+H]+ . 1 H NMR (400MHz, DMSO-d6) δ12.17(s,1H),11.07(s,1H),8.92(s,1H),8.36(d,J=8.0Hz,1H),7.90(s,1H),7.44(s,1H),7.13-7.03(m,1H),6.90-6. 68(m,3H),6.18-6.08(m,1H),5.74-5.66(m,1H),4.51-4.39(m,3H),3. 95(s,3H),3.71-3.61(m,2H),2.33-2.29(m,2H),1.52(d,J=8.0Hz,3H).

[0208] Example 21: 2-(2-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)thiazol-4-yl)-N-(4-(7-methyl-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide

[0209]

[0210] The preparation method is the same as that of Example 20, except that the raw material 7-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester is replaced with 7-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester to obtain the title compound. ESI-MS m / z: 567.2 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ11.99(s,1H),11.07(s,1H),8.93(s,1H),8.60(d, J=4.0Hz,1H),7.90(s,1H),7.44(s,1H),7.08-7.01(m,2H),6.94-6.76(m, 1H),6.72-6.68(m,1H),6.19-6.02(m,1H),5.75-5.57(m,1H),4.52-4.35( m,3H),3.67(s,2H),2.51(s,3H),2.33-2.29(m,2H),1.53(d,J=4.0Hz,3H).

[0211] 2-(2-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)thiazol-4-yl)-N-(4-(7-methyl-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide (420 mg) was separated by chiral preparative column. (Method: Chiral column: ChiralPak IG, 250×30 mm ID, 5 μm, elution: isocratic: B 40%, A=carbon dioxide, B=ethanol) to obtain Isomer 1 (163 mg) and Isomer 2 (182 mg) of the title compound.

[0212] Isomer 1 (Example 22): ESI-MS m / z: 567.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ11.99(s,1H),11.07(s,1H),8.92(s,1H),8.60(d,J=4.0Hz,1H),7.90(s,1H),7.45(s,1H),7.11-7.00(m,2H),6.91 -6.66(m,2H),6.19-6.02(m,1H),5.75-5.57(m,1H),4.52-4.35(m,3H),3.68(s,2H),2.51(s,3H),2.36-2.26(m,2H),1.53(d,J=4.0Hz,3H).

[0213] Isomer 2 (Example 23): ESI-MS m / z: 567.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ11.99(s,1H),11.07(s,1H),8.92(s,1H),8.60(d,J=4.0Hz,1H),7.90(s,1H),7.45(s,1H),7.11-7.00(m,2H),6.91 -6.66(m,2H),6.19-6.02(m,1H),5.75-5.57(m,1H),4.52-4.35(m,3H),3.68(s,2H),2.51(s,3H),2.36-2.26(m,2H),1.53(d,J=4.0Hz,3H).

[0214] Example 24: 2-(2-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)thiazol-4-yl)-N-(4-(6-trifluoromethyl-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide

[0215]

[0216] The preparation method is the same as that of Example 20, except that the raw material 7-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester is replaced with 6-trifluoromethyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester to obtain the title compound. ESI-MS m / z: 621.2 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ12.29(s,1H),11.17(s,1H),9.07-8.94(m,2H),8.20(s,1H),7.86(s,1H),7.51-7.39(m,2H),6.90-6.6 8(m,2H),6.19-6.04(m,1H),5.74-5.58(m,1H),4.53-4.37(m,3H),3.73-3.60(m,2H),2.36-2.26(m,2H),1.53(d,J=4.0Hz,3H).

[0217] 2-(2-(1-acryloyl-1,2,5,6-tetrahydropyridin-3-yl)thiazol-4-yl)-N-(4-(6-trifluoromethyl-1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)propanamide (155 mg) was separated by chiral preparation column. (Method: chiral column: (S,S) Whek O1, 250×30 mm ID, 10 μm, elution: isocratic: B 40%, A=carbon dioxide, B=ethanol) to obtain isomer 1 (40 mg) and isomer 2 (66 mg) of the title compound.

[0218] Isomer 1 (Example 25): ESI-MS m / z: 621.2 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ12.29(s,1H),11.17(s,1H),9.07-8.94(m,2H),8.20(s,1H),7.86(s,1H),7.51-7.39(m,2H),6.90-6.6 8(m,2H),6.19-6.04(m,1H),5.74-5.58(m,1H),4.53-4.37(m,3H),3.73-3.60(m,2H),2.36-2.26(m,2H),1.53(d,J=4.0Hz,3H).

[0219] Isomer 2 (Example 26): ESI-MS m / z: 621.2 [M+H] +. 1 H NMR(400MHz,DMSO-d6)δ12.29(s,1H),11.17(s,1H),9.07-8.94(m,2H),8.20(s,1H),7.86(s,1H),7.51-7.39(m,2H),6.90-6.6 8(m,2H),6.19-6.04(m,1H),5.74-5.58(m,1H),4.53-4.37(m,3H),3.73-3.60(m,2H),2.36-2.26(m,2H),1.53(d,J=4.0Hz,3H).

[0220] Example 27: N-(4-(1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)-2-(1'-acryloyl-1',2',5',6'-tetrahydro-[2,3'-bipyridyl]-5-yl)propanamide

[0221]

[0222] Step 1: Preparation of methyl 2-(1'-acryloyl-1',2',5',6'-tetrahydro-[2,3'-bipyridyl]-5-yl)propanoate

[0223]

[0224] Dissolve methyl 2-(6-chloropyridin-3-yl)propanoate (766 mg, 3.87 mmol, 1.0 eq) in 1,4-dioxane (90.0 mL), and add 1-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridin-1(2H)-yl)prop-2-en-1-one (1.21 g, 4.60 mmol, 1.2 eq), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (281 mg, 0.38 mmol, 0.1 eq), cesium carbonate (3.75 g, 11.52 mmol, 3.0 eq) and water (4.5 mL) in sequence, and reflux at 90 °C with stirring for 3 h under argon protection. The reaction was monitored by LC-MS. After the reaction was completed, water was added to quench the reaction, ethyl acetate was added to extract, the organic phases were combined, washed with water three times, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 690 mg of the title compound. ESI-MS m / z: 301.0 [M+H] + .

[0225] Step 2: Preparation of 2-(1'-acryloyl-1',2',5',6'-tetrahydro-[2,3'-bipyridyl]-5-yl)propanoic acid

[0226]

[0227] Dissolve methyl 2-(1'-acryloyl-1',2',5',6'-tetrahydro-[2,3'-bipyridyl]-5-yl)propanoate (255 mg, 0.85 mmol, 1.0 eq) in tetrahydrofuran (20 mL), add water (10 mL) and lithium hydroxide (136 mg, 3.4 mmol, 4.0 eq), and stir at room temperature for 2 h. LC-MS tracking and monitoring the reaction, after the reaction is completed, add water to quench, adjust the pH to 5 with dilute hydrochloric acid, add ethyl acetate to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate to obtain 153 mg of the title compound. ESI-MS m / z: 287.0 [M+H] + .

[0228] Step 3: Preparation of tert-butyl 3-(2-(2-(1-(1'-acryloyl-1',2',5',6'-tetrahydro-[2,3'-bipyridyl]-5-yl)propionamido)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-1-carboxylate

[0229]

[0230] Dissolve tert-butyl 3-(2-amino-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-1-carboxylate (98 mg, 0.26 mmol, 0.5 eq) in anhydrous dichloromethane (10 mL), add 2-(1'-acryloyl-1',2',5',6'-tetrahydro-[2,3'-bipyridyl]-5-yl)propionic acid (150 mg, 0.52 mmol, 1.0 eq) and pyridine (10 mL), cool to -20°C, add phosphorus oxychloride (80 mg, 0.52 mmol, 1.0 eq) dropwise, and stir for 2 h. LC-MS tracking and monitoring the reaction, after the reaction is completed, the reaction solution is poured into ice water, extracted with dichloromethane, and the organic phases are combined, washed three times with water, dried and concentrated over anhydrous sodium sulfate, and purified by column chromatography to obtain 53 mg of the title compound. ESI-MS m / z: 647.3[M+H] + .

[0231] Step 4: Preparation of N-(4-(1H-indol-3-yl)-5-(trifluoromethyl)pyrimidin-2-yl)-2-(1'-acryloyl-1',2',5',6'-tetrahydro-[2,3'-bipyridyl]-5-yl)propanamide

[0232]

[0233] 3-(2-(2-(1-(1'-acryloyl-1',2',5',6'-tetrahydro-[2,3'-bipyridine]-5-yl)propionamido)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-1-carboxylic acid tert-butyl ester (53 mg, 0.08 mmol, 1.0 eq) was dissolved in dichloromethane (3 mL), cooled to 0°C in an ice bath, trifluoroacetic acid (1 mL) was added dropwise, and the reaction was allowed to react at room temperature for 4 h. The reaction was monitored by LC-MS. After the reaction was completed, the reaction solution was dried and purified by C18 column chromatography to obtain 14 mg of the title compound. ESI-MS m / z: 546.9 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ11.98(s,1H),11.14(s,1H),9.00-8.74(m,2H),8.58(s,1H),7.97(s,1H),7.86-7.44(m,3H),7.32-7.14(m,2H),6.94 -6.70(m,2H),6.20-6.05(m,1H),5.76-5.63(m,1H),4.61-4.43(m,2H) ,4.32-4.20(m,1H),3.67(s,2H),2.42-2.25(m,2H),1.62-1.32(m,3H).

[0234] Example 28: 2-(1'-Acryloyl-1',2',5',6'-tetrahydro-[2,3'-bipyridyl]-5-yl)-N-(5-chloro-4-methoxypyrimidin-2-yl)-propionamide

[0235]

[0236] The preparation method is the same as that of Example 27, except that the raw material 3-(2-amino-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-1-carboxylic acid tert-butyl ester is replaced with 5-chloro-4-methoxypyrimidin-2-amine to obtain the title compound. ESI-MS m / z: 428.2 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ10.89(s,1H),8.54(s,1H),8.47(s,1H),7.76(d,J=8.0H z,1H),7.64(t,J=8.0Hz,1H),6.88(q,J=8.0Hz,1H),6.77(d,J=16.0Hz,1H),6.12 (q,J=8.0Hz,1H),5.72-5.69(m,1H),4.52(d,J=16.0Hz,2H),4.14(q,J=8.0Hz,1H ), 4.00 (s, 3H), 3.69-3.68 (m, 2H), 2.34 (d, J = 20.0Hz, 2H), 1.43 (d, J = 8.0Hz, 3H).

[0237] Example 29: 2-(1'-acryloyl-1',2',5',6'-tetrahydro-[2,3'-bipyridyl]-5-yl)-N-(1-methyl-6-oxo-1,6-dihydropyrimidin-2-yl)-propionamide

[0238]

[0239] Step 1: Preparation of N,N-dimethyl-N'-(6-oxo-1,6-dihydropyrimidin-2-yl)formamide

[0240]

[0241] Dissolve 2-aminopyrimidin-4(3H)-one (10 g, 0.08 mol, 1.0 eq) in anhydrous toluene (40 mL), add 1,1-dimethoxy-N,N-dimethylformamide (38.7 g, 0.32 mol, 4.0 eq), and stir at 60°C overnight. LC-MS was used to monitor the reaction. After the reaction was completed, the mixture was directly concentrated to obtain 12.4 g of the title compound. ESI-MS m / z: 167.1 [M+H] + .

[0242] Step 2: Preparation of N,N-dimethyl-N'-(1-methyl-6-oxo-1,6-dihydropyrimidin-2-yl)formamide

[0243]

[0244] Dissolve N,N-dimethyl-N'-(6-oxo-1,6-dihydropyrimidin-2-yl)formamide (11g, 0.066mol, 1.0eq) in anhydrous N,N-dimethylformamide (60mL), add potassium carbonate (10.95g, 0.079mol, 1.2eq) and iodomethane (11.3g, 0.079mol, 1.2eq) in turn, and stir at room temperature overnight. LC-MS tracking and monitoring the reaction, after the reaction is completed, add water to quench, add ethyl acetate to extract, combine the organic phases, wash the organic phases with water three times, dry and concentrate over anhydrous sodium sulfate, and purify by column chromatography to obtain 8.2g of the title compound. ESI-MS m / z: 181.1[M+H] + .

[0245] Step 3: Preparation of 2-amino-3-methylpyrimidin-4(3H)-one

[0246]

[0247] Dissolve N,N-dimethyl-N'-(1-methyl-6-oxo-1,6-dihydropyrimidin-2-yl)formamide (1g, 5.55mmol, 1.0eq) in ethanol (4mL) and place in a sealed tube. Add acetic acid (1g, 16.66mmol, 3.0eq) and hydrazine hydrate (1.82g, 36.1mmol, 6.5eq) in turn and stir at 50°C for 3h. LC-MS tracking and monitoring the reaction. After the reaction is completed, the solvent is dried and filtered. The filter cake is washed with dichloromethane to obtain 0.8g of the title compound. ESI-MS m / z: 126.1[M+H] + .

[0248] Step 4: Preparation of 2-(1'-acryloyl-1',2',5',6'-tetrahydro-[2,3'-bipyridyl]-5-yl)-N-(1-methyl-6-oxo-1,6-dihydropyrimidin-2-yl)-propionamide

[0249]

[0250] The preparation method is the same as that of Example 27, except that the raw material 3-(2-amino-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-1-carboxylic acid tert-butyl ester is replaced with 2-amino-3-methylpyrimidin-4(3H)-one to obtain the title compound. ESI-MS m / z: 394.2 [M+H] + . 1H NMR(400MHz, DMSO-d6)δ7.63(d,J=4.0Hz,1H),7.44-7.31(m,4H),7.00-6.80(m,1H),6.33-6.23(m,1H),6.18-6.10(m,1H),6.07-5.94(m ,1H),5.75-5.64(m,1H),4.48-4.32(m,2H),3.88-3.77(m,1H),3.73-3.60(m,2H),3.25(s,3H),2.35-2.2(m,2H),1.42(d,J=8.0Hz,3H).

[0251] Example 30: 2-(1'-acryloyl-1',2',5',6'-tetrahydro-[2,3'-bipyridine]-5-yl)-N-(quinazolin-2-yl)-propionamide

[0252]

[0253] The preparation method is the same as that of Example 27, except that the raw material 3-(2-amino-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-1-carboxylic acid tert-butyl ester is replaced with quinazolin-2-amine to obtain the title compound. ESI-MS m / z: 414.7 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.23(s,1H),8.62(s,1H),8.28(s,1H),7.98-7.73(m,3H),7.61-7.33(m,2H),6.84-6.52(m,2H) ,6.40-6.21(m,1H),5.70(d,J=8.0Hz,1H),4.85-4.40(m,3H),3.88-3.58(m,2H),2.40(s,2H),1.64(d,J=8.0Hz,3H).

[0254] Experimental Example 1: Evaluation of in vitro cell activity of compounds

[0255] 1. Experimental Materials

[0256] Test compound: The compounds of the present invention prepared in the above examples, each compound was prepared into a 20mM stock solution with DMSO, the stock solution was diluted with DMSO, the highest concentration was diluted 10 times to 2mM, and then diluted 4 times with DMSO in sequence to 500.00nM, 125.00nM, 31.25nM, 7.81nM, 1.95nM, 0.49nM, 0.12nM, 0.03nM. After the compound was diluted with DMSO, it was diluted 100 times with cell culture medium before administration, and the negative control well was culture medium plus DMSO of the same volume as the administration well.

[0257] The human breast cancer cell line MDA-MB-231 was purchased from the American Type Culture Collection (ATCC).

[0258] Reagents: RPMI-1640, purchased from Invitrogen, USA; L-15, purchased from Invitrogen, USA; FBS, purchased from Invitrogen, USA; pancreatin, purchased from Invitrogen, USA; P / S, purchased from Invitrogen, USA; DMSO, purchased from Sigma, USA; Luminescent Cell Viability Assay Kit was purchased from Progema, USA; Trypan blue staining solution was purchased from Progema, USA.

[0259] 2. Experimental Methods

[0260] 2.1 Cell culture:

[0261] Cell recovery: Dissolve the cells in a 37-degree water bath, then transfer to 15 mL of preheated culture medium, centrifuge at 1000 rpm for 5 min, discard the culture medium, resuspend the cells in 15 mL of fresh culture medium, transfer to a T75 culture flask, and culture in an incubator at 37°C, 5% CO2. After 24 hours, replace the cells with fresh culture medium.

[0262] Cell passaging: Transfer the above revived cells to a 15 mL sterile centrifuge tube, centrifuge at 1000 rpm for 5 min, discard the culture medium, mix with 1 mL of serum-containing culture medium, add to a 10 cm culture dish according to a certain ratio, and culture in an incubator at 37 degrees and 5% CO2.

[0263] 2.2 Experimental steps:

[0264] Day 0: Plating. After the cells grow to a confluence of 80%-90% in the culture flask, discard the culture medium in the dish, wash the dead cells with 1mL of PBS, discard the PBS, add 1mL of trypsin for digestion, and when the cells become brighter and rounder under the microscope, terminate the digestion with an equal volume of culture medium, gently blow off the cells with a gun tip, collect all the suspension in the dish into a centrifuge tube, centrifuge at 1000rpm for 5min, discard all the supernatant in the tube after centrifugation, mix the cells in the tube with 1mL of culture medium, and count them. According to the plating density of 3×10 3 Cells were plated into 96-well plates at 100 μL per well. The volume of culture medium + cells in each well was 100 μL. Four outer edges of the plate were filled with 200 μL PBS to prevent the culture medium at the edge from evaporating quickly, resulting in large differences in culture conditions in the inner wells. After plating, the cells were placed in culture medium for overnight culture.

[0265] Day 1: Dosing. The drug was prepared as described in the above “Test Compound”, and the drug diluted with culture medium was added to the 96-well plate prepared on the first day, 100 μL per well. The final drug concentrations in the plate were 10 μM, 2.5 μM, 0.625 μM, 0.15625 μM, 0.0390625 μM, 0.009765625 μM, 0.002441406 μM, 0.000610352 μM, 0.000152588 μM, 0 μM from left to right.

[0266] Day 3: Detection. After 3 days of drug treatment, the culture medium in the wells was aspirated and discarded. CellTiter-GloLuminescent Cell Viabillity Assay was taken out 30 minutes in advance and equilibrated to room temperature. Celltiter-Glo reagent was diluted 1:1 (equal volume) with 10% FBS culture medium, and 150 μL of diluted Celltiter-Glo reagent was added to each well, and shaken at room temperature for 2 minutes. After incubating at room temperature for 10 minutes, 100 μL was taken to detect the chemiluminescent signal, shaken, and the Read injection detection condition was 500ms.

[0267] Data processing and analysis

[0268] (1) According to the AU value derived from the microplate reader, calculate the inhibition rate of each well relative to the solvent control well:

[0269] Inhibition (%) = 100 - (AU experimental well - AU blank well) / (AU solvent control well - AU blank well) × 100

[0270] (2) According to different drug concentrations and their corresponding inhibition rates, IC was analyzed using GraphicPad 5.0 software. 50Draw curves, analyze data, and obtain final IC 50 value.

[0271] The experimental results are shown in Table 1.

[0272] Table 1

[0273]

[0274]

[0275] It can be seen from the above experiments that the compounds of the present invention exhibit good inhibitory activity against human breast cancer cells.

[0276] Although the present invention has been described in detail above, it is understood by those skilled in the art that various modifications and changes can be made to the present invention without departing from the spirit and scope of the present invention. The scope of the present invention is not limited to the detailed description above, but should be attributed to the claims.

Claims

1. A compound represented by general formula (I) or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, in, X1 and X2 are each independently selected from -CH- and -N-; R1, R2 and R3 are each independently selected from hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, oxygen, sulfur, thiol, alkylthio, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylalkylthio, heterocyclylalkylthio, arylalkylthio and heteroarylalkylthio, which are optionally substituted with one or more alkyl, alkoxy, alkylamino, halogen, hydroxy, amino, nitro, cyano, haloalkyl, alkylacyl, aminoacyl or alkylaminoacyl; or R1 and R2 together with the C atoms to which they are attached form cycloalkyl, heterocyclyl, aryl, heteroaryl, which are optionally substituted with one or more alkyl, alkoxy, alkylamino, halogen, hydroxy, amino, nitro, cyano, haloalkyl, alkylacyl, aminoacyl or alkylaminoacyl; R5 and R6 are each independently selected from hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, alkylthio, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino and oxo; R7 and R8 are each independently selected from hydrogen, halogen, hydroxy, carbonyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, alkylcarbonyl, alkenylcarbonyl, alkynylcarbonyl, optionally substituted with one or more alkyl, alkenyl, alkynyl, alkoxy, alkylamino, halogen, hydroxy, amino, haloalkyl, alkylacyl, aminoacyl or alkylaminoacyl; and is a single bond or a double bond, where When it is a double bond, R4 does not exist; When it is a single bond, R4 is selected from alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, alkylthio, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl and dialkylamino, and R3 is selected from hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, oxygen, sulfur, mercapto, alkylthio, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino, oxo, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylalkylthio, heterocyclylalkylthio, arylalkylthio and heteroarylalkylthio.

2. The compound according to claim 1 or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein R1, R2 and R3 are each independently selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, oxygen, sulfur, mercapto, C 1-6 Alkylthio, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, di-C 1-6 Alkylamino, C 3-8 Cycloalkyl, C 3-8 Heterocyclic group, C 6-12 Aryl, C 5-12 Heteroaryl, C 3-8 Cycloalkyl C 1-6 Alkylthio, C 3-8 Heterocyclyl C 1-6 Alkylthio, C 6-12 Aryl C 1-6 Alkylthio and C 5-12 Heteroaryl C 1-6 Alkylthio, optionally substituted with one or more C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, halogen, hydroxy, amino, nitro, cyano, halo C 1-3 Alkyl, C 1-3 Alkyl acyl, amino acyl or C 1-3 Alkylaminoacyl substitution.

3. The compound according to claim 1 or 2, or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, Wherein R1, R2 and R3 are each independently selected from It is optionally replaced by one or more C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, halogen, hydroxy, amino, nitro, cyano, halo C 1-6 Alkyl, C 1-6 Alkyl acyl, amino acyl or C 1-6 Alkylaminoacyl substitution.

4. The compound according to any one of claims 1 to 3 or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein R5 and R6 are each independently selected from hydrogen, halogen, hydroxyl, alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, C 1-6 Alkylthio, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, di-C 1-6 Alkylamino and oxo groups.

5. The compound according to any one of claims 1 to 4, or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein R7 and R8 are each independently selected from hydrogen, halogen, hydroxyl, carbonyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, C 1-6 Alkylcarbonyl, C 2-6 Alkenylcarbonyl, C 2-6 Alkynylcarbonyl, which is optionally substituted with one or more C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Alkylamino, halogen, hydroxy, amino, halo C 1-6 Alkyl, C 1-6 Alkyl acyl, amino acyl or C 1-6 Alkylaminoacyl substitution.

6. A compound represented by general formula (II) or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, in, X1, X2, R1, R2, R3, R4, R5, R6, R7, R8 and As defined in the general formula (I) in claims 1-5.

7. A compound represented by general formula (III) or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, in, X1, X2, R1, R2, R3, R4, R5, R6, R7, R8 and As defined in the general formula (I) in claims 1-5.

8. The compound according to any one of claims 1 to 7 or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein the compound is selected from the following compounds:

9. A pharmaceutical composition comprising the compound according to any one of claims 1 to 8 or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug and a pharmaceutically acceptable carrier.

10. Use of the compound according to any one of claims 1 to 8 or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, or the pharmaceutical composition according to claim 9 in the preparation of a medicament for treating a disease associated with CDK12 / 13.

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