A pyrazolopyrimidine derivative, a process for its preparation and use thereof

Abstract

The present application provides a compound of Formula I, isomers thereof, or pharmaceutically acceptable salts thereof, and methods of making and medical uses thereof. The compounds are useful as pharmaceutical agents in the treatment of diseases and conditions, including cancer or tumors, and the like.

Description

Technical Field

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

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

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

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

[0005] One object of this application is to provide pyrazolopyrimidine derivatives, isomers thereof, or pharmaceutically acceptable salts thereof with the structure shown in Formula I:

[0006]

[0007] in,

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

[0009] Ring B is selected from saturated cycloalkyl, saturated heterocycloalkyl, unsaturated cycloalkyl or unsaturated heterocycloalkyl, wherein the cycloalkyl or heterocycloalkyl can be selected from monocyclic, bicyclic, bridged or spirocyclic rings;

[0010] R 1 Selected from: hydrogen, halogen, cyano, nitro, oxo, hydroxyl, substituted or unsubstituted amino groups, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkyloxy or heteroaryl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkyloxy or heteroaryl is unsubstituted or substituted by one or more halogens, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy;

[0011] R 2 Selected from: hydrogen, halogen, cyano, oxo, hydroxyl, substituted or unsubstituted amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy is unsubstituted or substituted by one or more halogens, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy;

[0012] R 3 Selected from hydrogen, halogen, cyano, substituted or unsubstituted amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy is unsubstituted or substituted by one or more halogens, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy;

[0013] R 4 R 5 Each is independently selected from: hydrogen, C1-C6 alkyl, or C3-C8 cycloalkyl;

[0014] or R 4 With R 5 It forms a ring with the atoms it is attached to;

[0015] m is selected from: 1, 2, 3, 4 or 5;

[0016] n is selected from: 1, 2, 3, 4 or 5.

[0017] Preferably, the compound of formula I, its isomers, or pharmaceutically acceptable salts thereof,

[0018] Ring A is selected from 5-10 member unsaturated monocyclic or bicyclic heterocyclic hydrocarbon groups, wherein the heterocyclic hydrocarbon group contains at least one nitrogen atom as a heteroatom;

[0019] Cycle B is selected from 4-8 saturated cycloalkyl or 4-10 saturated heterocycloalkyl, wherein the heterocycloalkyl contains at least one nitrogen atom as a heteroatom, and wherein the cycloalkyl or heterocycloalkyl can be selected from monocyclic, bicyclic, bridged or spirocyclic.

[0020] R 1 Selected from: hydrogen, halogen, cyano, nitro, oxo, hydroxyl, substituted or unsubstituted amino groups, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkyloxy or heteroaryl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkyloxy or heteroaryl is unsubstituted or substituted by one or more halogens, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy;

[0021] R 2 Selected from: hydrogen, halogen, cyano, oxo, hydroxyl, substituted or unsubstituted amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy is unsubstituted or substituted by one or more halogens, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy;

[0022] R 3 Selected from hydrogen, halogen, cyano, substituted or unsubstituted amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy is unsubstituted or substituted by one or more halogens, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy;

[0023] R 4 R 5 Each is independently selected from: hydrogen, C1-C6 alkyl, or C3-C8 cycloalkyl;

[0024] or R 4 With R 5 It forms a ring with the atoms it is attached to;

[0025] m is selected from: 1, 2, 3, 4 or 5;

[0026] n is selected from: 1, 2, 3, 4 or 5.

[0027] Preferably, the compound of formula I, its isomers, or pharmaceutically acceptable salts thereof,

[0028] Ring A is selected from 5-10 membered heterocyclic aryl groups, wherein the heterocyclic aryl group contains at least one nitrogen atom as a heteroatom;

[0029] Cycle B is selected from 5-8 membered saturated heterocyclic alkyl groups, wherein the heterocyclic alkyl group contains at least one nitrogen atom as a heteroatom;

[0030] R 1 Selected from: hydrogen, halogen, cyano, nitro, oxo, hydroxyl, substituted or unsubstituted amino groups, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkyloxy or heteroaryl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkyloxy or heteroaryl is unsubstituted or substituted by one or more halogens, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy;

[0031] R 2 Selected from: hydrogen, halogen, cyano, oxo, hydroxyl, substituted or unsubstituted amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy is unsubstituted or substituted by one or more halogens, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy;

[0032] R 3 Selected from hydrogen, halogen, cyano, substituted or unsubstituted amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy is unsubstituted or substituted by one or more halogens, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy;

[0033] R 4 R 5 Each is independently selected from: hydrogen, C1-C6 alkyl, or C3-C8 cycloalkyl;

[0034] or R 4 With R 5 It forms a ring with the atoms it is attached to;

[0035] m is selected from: 1, 2, 3, 4 or 5;

[0036] n is selected from: 1, 2, 3, 4 or 5.

[0037] Preferably, the compound of formula I, its isomers, or pharmaceutically acceptable salts thereof further have the structure shown in formula II.

[0038]

[0039] in,

[0040] Y 1 Y 2 Y 3 Y 4 Y 5 Selected independently from: C, N, CR 1 or NR 6 ;

[0041] R 1 Selected from: hydrogen, halogen, cyano, nitro, oxo, hydroxyl, substituted or unsubstituted amino groups, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkyloxy or heteroaryl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkyloxy or heteroaryl is unsubstituted or substituted by one or more halogen, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy substituents;

[0042] R 2 Selected from: hydrogen, halogen, cyano, oxo, hydroxyl, substituted or unsubstituted amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy is unsubstituted or substituted by one or more halogens, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy;

[0043] R 3 Selected from hydrogen, halogen, cyano, substituted or unsubstituted amino, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy is unsubstituted or substituted by one or more halogens, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy;

[0044] R 4 R 5 Each is independently selected from: hydrogen, C1-C6 alkyl, or C3-C8 cycloalkyl;

[0045] or R 4 With R 5 It forms a ring with the atoms it is attached to;

[0046] R 6 Selected from: hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl or heteroaryl, wherein the C1-C6 alkyl, C3-C8 cycloalkyl or heteroaryl is unsubstituted or substituted by one or more halogen, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy substituents;

[0047] m is selected from: 1, 2, or 3;

[0048] n is selected from: 1 or 2.

[0049] More preferably, the compounds are pyrazolopyrimidine derivatives of Formula I, their isomers, or pharmaceutically acceptable salts thereof, selected from:

[0050]

[0051] Another objective of this application is to provide a method for preparing pyrazolopyrimidine derivatives with the structure shown in Formula I above, the method comprising the following steps:

[0052]

[0053] Where LG and LG' represent leaving groups, and R 1 R 2 R 3 Ring A, ring B, m, and n are defined above for pyrazolopyrimidine derivatives with the structure shown in Formula I;

[0054] (1) React compound I-1 with compound I-2 to obtain I-3;

[0055] The reaction is preferably carried out in a suitable organic solvent; the organic solvent may be selected from tetrahydrofuran, N,N-dimethylformamide, N,N-dimethylacetamide, 1,4-dioxane, acetonitrile, and any combination thereof, with acetonitrile being preferred. The reaction is preferably carried out in the presence of a suitable base. The base may be selected from triethylamine, pyridine, 4-dimethylaminopyridine, diisopropylethylamine, potassium carbonate, cesium carbonate, and sodium carbonate, with triethylamine being preferred. The reaction is preferably carried out at a suitable temperature, preferably 50-80°C. The reaction is preferably carried out for a suitable time, for example, 2-8 hours.

[0056] (2) The derivative with the structure shown in Formula I is obtained by coupling reaction of compound I-3 with boric acid or boric ester of compound I-4;

[0057] The reaction is preferably carried out in a suitable organic solvent. The organic solvent may be selected from tetrahydrofuran, 1,4-dioxane, toluene, acetonitrile, ethanol, water, and any combination thereof, preferably a combination of 1,4-dioxane and water. The reaction is preferably carried out in the presence of a suitable catalyst. The catalyst may be selected from Pd(dppf)Cl2, Pd(OAc)2, Pd2(dba)3, Pd(PPh3)4, preferably Pd(dppf)Cl2. The base may be selected from triethylamine, pyridine, 4-dimethylaminopyridine, diisopropylethylamine, potassium carbonate, cesium carbonate, sodium carbonate, preferably potassium carbonate. The reaction is preferably carried out at a suitable temperature, preferably 80-120°C. The reaction is preferably carried out for a suitable time, for example, 8-12 hours.

[0058] The specific conditions for each of the above reaction steps are well known in the art, and this invention does not specifically limit them. If the teachings of this invention are combined with common knowledge in the art, those skilled in the art can selectively replace the substituents in the general formula to prepare different compounds, and such selections and substitutions are all within the scope of protection of this invention.

[0059] This application also relates to the use of the compound of Formula I in the preparation of medicaments for the prevention or treatment of diseases related to CDK7.

[0060] In some embodiments, the CDK7-related diseases or conditions are selected from tumors or cancers, such as breast cancer, pancreatic cancer, ovarian cancer, colorectal cancer, lung cancer, prostate cancer, lymphoma, malignant sarcoma, cervical cancer, oral cancer, brain cancer, stomach cancer, liver cancer, skin cancer, bone cancer, kidney cancer, bladder cancer, fallopian tube tumors, peritoneal tumors, melanoma, glioma, glioblastoma, papillary malignant tumors, head and neck tumors, myeloma, or leukemia.

[0061] This application discovers a novel class of CDK7 inhibitors with the structure shown in Formula I, which exhibits good CDK7 enzyme inhibitory activity. Detailed Implementation

[0062] To make the objectives and technical solutions of this application clearer, the present invention is further described below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of this application. Furthermore, specific experimental methods not mentioned in the following embodiments are all performed according to conventional experimental methods.

[0063] In this application, when the chemical name and structural formula are inconsistent, the structural formula shall prevail, unless the chemical name rather than the structural formula can be inferred to be correct from the context.

[0064] The abbreviations used in this article have the following meanings:

[0065] abbreviation meaning <![CDATA[DMSO-d6]]> Hexadecimalized dimethyl sulfoxide TMS Tetramethylsilane <![CDATA[ 1 H NMR]]> Hydrogen spectrum MS mass spectrometry s Single peak d Double peak t Triple Peak q Four Peaks dd Double peak m Multiplets br Broad Peak J Coupling constant Hz hertz

[0066] The structure of the compound was determined by mass spectrometry (MS) or nuclear magnetic resonance (NMR). 1 It was determined by H NMR.

[0067] Nuclear magnetic resonance (NMR) 1 H NMR shift (δ) is given in parts per million (ppm); nuclear magnetic resonance (NMR) 1 The 1H NMR (hydrogen spectroscopy) measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvent was deuterated dimethyl sulfoxide (DMSO-d6), and the internal standard was tetramethylsilane (TMS). Chemical shifts were expressed as 10⁻⁶ ppm. -6 (ppm) is given as the unit.

[0068] Mass spectrometry (MS) measurements were performed using a FINNIGAN LCQAd (ESI) mass spectrometer (manufacturer: Therm, model: Finnigan LCQ advantage MAX).

[0069] Thin-layer silicone uses Yantai Huanghai HSGF254 or Qingdao GF254 silicone sheets.

[0070] Column chromatography typically uses Yantai Huanghai silica gel with a mesh size of 200-300 as the carrier.

[0071] Unless otherwise specified, all reactions mentioned in this invention are carried out under a nitrogen atmosphere.

[0072] In the terminology of this invention, "nitrogen atmosphere" refers, for example, connecting a reaction vessel to a 1L nitrogen balloon.

[0073] In the terminology of this invention, "hydrogen atmosphere" refers, for example, connecting a reaction vessel to a 1L hydrogen balloon.

[0074] Unless otherwise specified in this invention, the solution mentioned in the reaction of this invention is an aqueous solution.

[0075] In the terminology of this invention, "room temperature" refers to a temperature between 10°C and 25°C.

[0076] Example 1 Preparation of (S)-N-(6-cyano-3-(3-cyano-7-(piperidin-3-amino)pyrazol[1,5-a]pyrimidin-5-yl)-1H-indole-7-yl)acetamide (1)

[0077]

[0078] Step 1: Preparation of tert-butyl 7-bromo-6-cyano-1H-indole-1-carboxylic acid (1b)

[0079] Compound 1a (500 mg, 2.26 mmol) and di-tert-butyl dicarbonate (332 mg, 2.71 mmol) were dissolved in dichloromethane (20 mL). 4-Dimethylaminopyridine (99 mg, 0.45 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 653 mg of the title compound (yield: 90%).

[0080] Step 2: Preparation of tert-butyl 7-acetamido-6-cyano-1H-indole-1-carboxylic acid (1c)

[0081] Compound 1b (500 mg, 1.56 mmol), acetamide (276 mg, 4.67 mmol), Pd2(dba)3 (90 mg, 0.16 mmol), XantPhos (180 mg, 0.31 mmol), and Cs2CO3 (1.01 g, 4.67 mmol) were dissolved in dry 1,4-dioxane (20 mL). The reaction mixture was heated to 110 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 117 mg of the title compound, yield: 25%.

[0082] Step 3: Preparation of tert-butyl 7-acetamido-3-bromo-6-cyano-1H-indole-1-carboxylic acid (1d)

[0083] Compound 1c (110 mg, 0.37 mmol) and NBS (85 mg, 0.48 mmol) were dissolved in tetrahydrofuran (4 mL), and the reaction mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 97 mg of the title compound, yield: 70%.

[0084] Step 4: Preparation of tert-butyl 7-acetamido-6-cyano-3-(4,4,5,5-tetramethyl-1,3,2-dioxoron-2-yl)-1H-indole-1-carboxylic acid (1e).

[0085] Compound 1d (95 mg, 0.25 mmol), pinacol diborate (191 mg, 0.75 mmol), Pd(dppf)Cl2 (27 mg, 0.04 mmol), and potassium acetate (74 mg, 0.75 mmol) were dissolved in a dry 1,4-dioxane solution (5 mL). Under nitrogen protection, the reaction mixture was heated to 110 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 53 mg of the title compound (yield: 50%).

[0086] Step 5: Preparation of (S)-3-((5-chloro-3-cyanopyrazolo[1,5-a]pyrimidin-7-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (1 g)

[0087] Compound 1f (550 mg, 2.58 mmol) and (S)-1-Boc-3-aminopiperidine (620 mg, 3.10 mmol) were dissolved in acetonitrile (13 mL), and then triethylamine (522 mg, 5.16 mmol) was added. The reaction mixture was stirred at 50 °C for 4 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 875 mg of the title compound, yield: 90%.

[0088] Step 6: Preparation of (S)-7-acetamido-3-(7-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-3-cyanopyrazolo[1,5-a]pyrimidin-5-yl)-6-cyano-1H-indole-1-carboxylic acid tert-butyl ester (1h)

[0089] 1 g (50 mg, 0.13 mmol) of compound, 1e (56 mg, 0.13 mmol), Pd(dppf)Cl2 (10 mg, 0.013 mmol), and K2CO3 (37 mg, 0.27 mmol) were dissolved in 1,4-dioxane / water (4 mL / 0.5 mL). The reaction mixture was heated to 110 °C and stirred overnight under nitrogen protection. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 25 mg of the title compound (yield: 30%).

[0090] Step 7: Preparation of (S)-N-(6-cyano-3-(3-cyano-7-(piperidin-3-amino)pyrazol[1,5-a]pyrimidin-5-yl)-1H-indole-7-yl)acetamide (1)

[0091] Compound 1h (25 mg, 39 μmol) was dissolved in dichloromethane (2 mL), and then hydrochloric acid-ethyl acetate solution (2 mL, 4 M) was added. The reaction system was stirred at room temperature for 2 hours. The reaction system was concentrated under reduced pressure, diluted with water, and the pH of the system was adjusted to about 8 with saturated sodium bicarbonate solution. The system was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel plate to give 7 mg of the title compound, yield: 40%.

[0092] LC-MS(ESI)m / z(M+H) + : 440.2

[0093] 1H NMR (400MHz, DMSO-d6) δ12.30(s,1H),10.29(s,1H),8.91(d,J=8.4Hz,1H),8.64(d,J=3.4 Hz,1H),8.13(s,1H),7.96(s,1H),7.30(dd,J=8.3,3.5Hz,1H),6.99(s,1H),3.96(s,1H), 3.04–3.00(m,1H),2.81–2.78(m,1H),2.71–2.66(m,1H),2.58–2.55(m,1H),2.47–2.44(m ,1H),2.06(s,3H),1.88–1.86(m,1H),1.82–1.72(m,1H),1.54–1.52(m,1H),1.22(s,1H).

[0094] Example 2 Preparation of (S)-5-(6-cyano-7-((3-methyloxetane-3-yl)amino)-1H-indol-3-yl)-7-(piperidine-3-amino)pyrazol[1,5-a]pyrimidine-3-carboxynitrile (2)

[0095]

[0096] Step 1: Preparation of 6-cyano-7-((3-methyloxetane-3-yl)amino)-1H-indole-1-carboxylic acid tert-butyl ester (2a)

[0097] Compound 1b (500 mg, 1.56 mmol), 3-methyl-3-aminooxetane (407 mg, 4.67 mmol), Pd2(dba)3 (90 mg, 0.16 mmol), XantPhos (180 mg, 0.31 mmol), and Cs2CO3 (1.01 g, 4.67 mmol) were dissolved in a dry 1,4-dioxane (20 mL). The reaction mixture was heated to 110 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 142 mg of the title compound, yield: 28%.

[0098] Step 2: Preparation of tert-butyl 3-bromo-6-cyano-7-((3-methyloxetane-3-yl)amino)-1H-indole-1-carboxylic acid (2b)

[0099] Compound 2a (130 mg, 0.40 mmol) and NBS (92 mg, 0.52 mmol) were dissolved in tetrahydrofuran (4 mL). The reaction mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 113 mg of the title compound, yield: 70%.

[0100] Step 3: Preparation of 6-cyano-7-((3-methyloxetane-3-yl)amino)-3-(4,4,5,5-tetramethyl-1,3,2-dioxoborane-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester (2c)

[0101] Compound 2b (100 mg, 0.25 mmol), pinacol diborate (187 mg, 0.74 mmol), Pd(dppf)Cl2 (27 mg, 0.04 mmol), and potassium acetate (72 mg, 0.74 mmol) were dissolved in a dry 1,4-dioxane solution (5 mL). The reaction mixture was heated to 110 °C and stirred overnight under nitrogen protection. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 56 mg of the title compound (yield: 50%).

[0102] Step 4: Preparation of (S)-3-(7-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-3-cyanopyrazolo[1,5-a]pyrimidin-5-yl)-6-cyano-7-((3-methyloxetane-3-yl)amino)-1H-indole-1-carboxylic acid tert-butyl ester (2d)

[0103] 1 g (45 mg, 0.12 mmol) of compound 2c (54 mg, 0.12 mmol), Pd(dppf)Cl2 (9 mg, 0.012 mmol), and potassium carbonate (33 mg, 0.24 mmol) were dissolved in 1,4-dioxane / water (4 mL / 0.5 mL). The reaction mixture was heated to 110 °C and stirred overnight under nitrogen protection. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 24 mg of the title compound (yield: 30%).

[0104] Step 5: Preparation of (S)-5-(6-cyano-7-((3-methyloxetane-3-yl)amino)-1H-indol-3-yl)-7-(piperidin-3-amino)pyrazol[1,5-a]pyrimidine-3-carboxynitrile (2)

[0105] Compound 2d (24 mg, 36 μmol) was dissolved in DCM (2 mL), and then hydrochloric acid-ethyl acetate solution (2 mL, 4 M) was added. The reaction system was stirred at room temperature for 2 hours. The reaction system was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel filtration to obtain 9 mg of the title compound, yield: 55%.

[0106] LC-MS(ESI)m / z(M+H) + : 468.2

[0107] 1 H NMR (400MHz, DMSO-d6) δ11.28(s,1H),8.94(d,J=8.4Hz,1H),8.88(s,1H),8.54(d,J=3.4Hz,1H),8 .21(s,1H),7.84(s,1H),7.03(dd,J=8.3,3.5Hz,1H),6.77(s,1H),5.01–4.97(m,2H),4.76–4.72(m ,2H),4.01(s,1H),3.02–2.98(m,1H),2.83–2.79(m,1H),2.72–2.68(m,1H),2.60–2.58(m,1H),2.4 7–2.43(m,1H),1.88–1.86(m,1H),1.82–1.72(m,1H),1.54–1.52(m,1H),1.35(s,3H),1.22(s,1H).

[0108] Example 3 Preparation of (S)-5-(1H-indol-3-yl)-7-(piperidin-3-ylamino)pyrazolo[1,5-a]pyrimidine-3-carboxylonitrile (3)

[0109]

[0110] Step 1: Preparation of tert-butyl(S)-3-(7-((1-(tert-butyloxycarbonyl)piperidin-3-yl)amino)-3-cyanopyrazolo[1,5-a]pyrimidin-5-yl)-1H-indole-1-carboxylate (3a)

[0111] 1 g (100 mg, 0.27 mmol) of the compound, (1-(tert-butoxycarbonyl)-1H-indol-3-yl)boronic acid (104 mg, 0.40 mmol), Pd(dppf)Cl2 (19 mg, 0.027 mmol), and potassium carbonate (73 mg, 0.53 mmol) were dissolved in 1,4-dioxane / water (4 mL / 0.5 mL). Under nitrogen protection, the reaction mixture was heated to 110 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 66 mg of the title compound, yield: 45%.

[0112] Step 2: Preparation of (S)-5-(1H-indol-3-yl)-7-(piperidin-3-ylamino)pyrazolo[1,5-a]pyrimidine-3-carboxylonitrile (3)

[0113] Compound 3a (60 mg, 36 μmol) was dissolved in dichloromethane (2 mL), and then hydrochloric acid-ethyl acetate solution (2 mL, 4 M) was added. The reaction system was stirred at room temperature for 2 hours. The reaction system was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 27 mg of the title compound, yield: 70%.

[0114] LC-MS(ESI)m / z(M+H) + 358.2

[0115] 1 H NMR (400MHz, DMSO-d6) δ11.98(d,J=2.9Hz,1H),8.74(d,J=2.9Hz,1H),8.71–8.65(m,1H),8.58(s,1H),8.11(d,J=8.7Hz,1H ),7.50–7.43(m,1H),7.25–7.15(m,3H),4.41(s,1H),3.34–3.24(m,2H),2.86–2.76(m,2H),2.05–1.82(m,4H),1.22(s,1H).

[0116] Example 4 Preparation of (S)-3-ethyl-5-(1H-indol-3-yl)-N-(piperidin-3-yl)pyrazolo[1,5-a]pyrimidine-7-amine (4)

[0117]

[0118] Step 1: Preparation of (S)-3-((5-chloro-3-ethylpyrazolo[1,5-a]pyrimidin-7-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (4b)

[0119] Compound 4a (500 mg, 2.31 mmol) and (S)-1-Boc-3-aminopiperidine (556 mg, 2.78 mmol) were dissolved in acetonitrile (13 mL), and then triethylamine (468 mg, 4.63 mmol) was added. The reaction mixture was stirred at 50 °C for 4 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 791 mg of the title compound, yield: 90%.

[0120] Step 2: Preparation of (S)-3-(7-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-3-ethylpyrazolo[1,5-a]pyrimidin-5-yl)-1H-indole-1-carboxylic acid tert-butyl ester (4c)

[0121] Compound 4b (103 mg, 0.27 mmol), (1-(tert-butoxycarbonyl)-1H-indol-3-yl)boronic acid (104 mg, 0.40 mmol), Pd(dppf)Cl2 (19 mg, 0.027 mmol), and potassium carbonate (73 mg, 0.53 mmol) were dissolved in 1,4-dioxane / water (4 mL / 0.5 mL). Under nitrogen protection, the reaction mixture was heated to 110 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 53 mg of the title compound (yield: 35%).

[0122] Step 3: Preparation of (S)-3-ethyl-5-(1H-indol-3-yl)-N-(piperidin-3-yl)pyrazolo[1,5-a]pyrimidine-7-amine (4)

[0123] Compound 4c (53 mg, 94.5 μmol) was dissolved in dichloromethane (2 mL), and then hydrochloric acid-ethyl acetate solution (2 mL, 4 M) was added. The reaction system was stirred at room temperature for 2 hours. The reaction system was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 22 mg of the title compound, yield: 65%.

[0124] LC-MS(ESI)m / z(M+H) + 361.2

[0125] 1 H NMR(400MHz,DMSO-d6)δ11.69(s,1H),8.77–8.64(m,1H),8.30(s,1H),7.87(s,1H),7.51–7 .40(m,1H),7.22–7.12(m,2H),7.07(d,J=9.0Hz,1H),6.60(s,1H),3.87–3.84(m,1H),3.06– 3.02(m,1H),2.81–2.75(m,3H),2.71–2.68(m,1H),2.62–2.57(m,1H),1.88–1.85(m,1H),1 .77–1.74(m,1H),1.66–1.61(m,1H),1.53–1.49(m,1H),1.35(d,J=7.5Hz,3H),1.22(s,1H).

[0126] Example 5 Preparation of (S)-5-(6-cyano-7-dimethylphospho)-1H-indol-3-yl)-7-(piperidin-3-amino)pyrazolo[1,5-a]pyrimidine-3-carboxynitrile (5)

[0127]

[0128] Step 1: Preparation of 6-cyano-7-dimethyloxophosphoro-1H-indole (5a)

[0129] Compound 1a (1 g, 4.5 mmol) was dissolved in N,N-dimethylformamide (10 mL), and XantPhos (261 mg, 0.45 mmol), Pd2(dba)3 (206 mg, 0.225 mmol), K3PO4 (1.05 g, 4.95 mmol), and dimethylphosphorus oxide (422 mg, 5.4 mmol) were added sequentially. The reaction mixture was microwaved at 150 °C for 1 hour under nitrogen protection. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 393 mg of the title compound (yield: 40%).

[0130] Step 2: Preparation of 3-bromo-6-cyano-7-dimethyloxophosphoro-1H-indole (5b)

[0131] Compound 5a (393 mg, 1.8 mmol) and NBS (414 mg, 2.34 mmol) were dissolved in tetrahydrofuran (5 mL). The reaction mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 374 mg of the title compound, yield: 70%.

[0132] Step 3: Preparation of 6-cyano-7-dimethyloxophosphoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxorane-2-yl)-1H-indole (5c)

[0133] Compound 5b (393 mg, 1.26 mmol), pinacol diborate (955 mg, 3.78 mmol), Pd(dppf)Cl2 (48 mg, 0.07 mmol), and potassium acetate (368 mg, 3.78 mmol) were dissolved in 10 mL of dry 1,4-dioxane. The reaction mixture was heated to 110 °C and stirred overnight under nitrogen protection. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 138 mg of the title compound (yield: 32%).

[0134] Step 4: Preparation of (S)-3-(7-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-3-cyanopyrazolo[1,5-a]pyrimidin-5-yl)-6-cyano-7-dimethyloxophosphoryl-1H-indole (5d)

[0135] 1 g (100 mg, 0.27 mmol) of compound 5c (138 mg, 0.4 mmol), 19 mg (0.027 mmol) of Pd(dppf)Cl2, and 73 mg (0.53 mmol) of potassium carbonate were dissolved in 1,4-dioxane / water (4 mL / 0.5 mL). The reaction mixture was heated to 110 °C and stirred overnight under nitrogen protection. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 63 mg of the title compound (yield: 42%).

[0136] Step 5: Preparation of (S)-5-(6-cyano-7-dimethylphospho)-1H-indol-3-yl)-7-(piperidin-3-amino)pyrazolo[1,5-a]pyrimidine-3-carboxynitrile (5)

[0137] Compound 5d (63 mg, 112.8 μmol) was dissolved in dichloromethane (2 mL), and then hydrochloric acid-ethyl acetate solution (2 mL, 4 M) was added. The reaction system was stirred at room temperature for 2 hours. The reaction system was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 26 mg of the title compound, yield: 50%.

[0138] LC-MS(ESI)m / z(M+H) + : 459.2

[0139] 1 H NMR (400MHz, DMSO-d6) δ12.29(s,1H),8.89(d,J=8.4Hz,1H),8.58(d,J=3.4Hz,1H),8.1 4(s,1H),7.69(s,1H),7.50(dd,J=8.3,3.5Hz,1H),6.95(s,1H),3.96(s,1H),3.03–3.0 0(m,1H),2.80–2.75(m,1H),2.71–2.66(m,1H),2.58–2.55(m,1H),2.47–2.43(m,1H),1 .98–1.95(m,6H),1.88–1.86(m,1H),1.82–1.72(m,1H),1.54–1.52(m,1H),1.22(s,1H).

[0140] Example 6 Preparation of (S)-5-(1H-pyrrolo[2,3-b]pyridin-3-yl)-7-(piperidine-3-amino)pyrazolo[1,5-a]pyrimidine-3-carboxylonitrile (6)

[0141]

[0142] Step 1: Preparation of (S)-3-(7-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-3-cyanopyrazolo[1,5-a]pyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridine (6a)

[0143] 1 g (200 mg, 0.54 mmol) of the compound, 3-(4,4,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)-1H-pyrrolo[2,3-b]pyridine (195 mg, 0.80 mmol), Pd(dppf)Cl2 (38 mg, 0.054 mmol), and potassium carbonate (146 mg, 1.06 mmol) were dissolved in 1,4-dioxane / water (8 mL / 0.5 mL). Under nitrogen protection, the reaction mixture was heated to 110 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 70 mg of the title compound, yield: 38%.

[0144] Step 2: Preparation of (S)-5-(1H-pyrrolo[2,3-b]pyridin-3-yl)-7-(piperidine-3-amino)pyrazolo[1,5-a]pyrimidine-3-carboxylonitrile (6)

[0145] Compound 6a (70 mg, 152.7 μmol) was dissolved in dichloromethane (4 mL), and then hydrochloric acid-ethyl acetate solution (2 mL, 4 M) was added. The reaction system was stirred at room temperature for 2 hours. The reaction system was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 26 mg of the title compound, yield: 48%.

[0146] LC-MS(ESI)m / z(M+H) + 359.2

[0147] 1H NMR (400MHz, DMSO-d6) δ9.94(d,J=10.4Hz,1H),9.63(d,J=10.2Hz,1H),8.72(d,J=8.7Hz,1 H),8.58(dd,J=4.8,1.6Hz,1H),8.44(d,J=4.0Hz,1H),8.15(dd,J=7.8,1.6Hz,1H),7.34(dd ,J=7.8,4.8Hz,1H),6.85(d,J=4.0Hz,1H),4.23(s,1H),3.48–3.44(m,1H),3.26–3.23(m,1H ),3.12–3.04(m,1H),2.81–2.78(m,1H),2.16–2.12(m,1H),1.99–1.81(m,3H),1.22(s,1H).

[0148] Example 7 Preparation of (S)-5-(6-cyano-1H-indol-3-yl)-7-(piperidin-3-amino)pyrazole[1,5-a]pyrimidine-3-carboxynitrile (7)

[0149]

[0150] Step 1: Preparation of (S)-3-(7-((1-(tert-butoxycarbonyl)piperidin-3-yl)amino)-3-cyanopyrazolo[1,5-a]pyrimidin-5-yl)-6-cyano-1H-indole-1-carboxylic acid tert-butyl ester (7a)

[0151] 1 g (200 mg, 0.54 mmol) of the compound, 295 mg (0.80 mmol) of 6-cyano-3-(4,4,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)-1H-indole, 38 mg (0.054 mmol) of Pd(dppf)Cl2, and 146 mg (1.06 mmol) of potassium carbonate were dissolved in 1,4-dioxane / water (8 mL / 0.5 mL). The reaction mixture was heated to 110 °C and stirred overnight under nitrogen protection. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 65 mg of the title compound (yield: 21%).

[0152] Step 2: Preparation of (S)-5-(6-cyano-1H-indol-3-yl)-7-(piperidin-3-amino)pyrazole[1,5-a]pyrimidine-3-carboxynitrile (7)

[0153] Compound 7a (65 mg, 111.6 μmol) was dissolved in dichloromethane (4 mL), and then hydrochloric acid-ethyl acetate solution (2 mL, 4 M) was added. The reaction system was stirred at room temperature for 2 hours. The reaction system was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 25 mg of the title compound, yield: 59%.

[0154] LC-MS(ESI)m / z(M+H) + 383.2

[0155] 1 H NMR (400MHz, DMSO-d6) δ12.24 (s, 1H), 8.82–8.75 (m, 2H), 8.58 (s, 1H), 8.04 (d, J = 1. 4Hz,1H),7.78(d,J=1.8Hz,1H),7.55(dd,J=8.4,1.5Hz,1H),6.99(s,1H),3.92(s,1H ),3.04–3.02(m,1H),2.81–2.78(m,1H),2.71–2.66(m,1H),2.56–2.52(m,1H),1.90 –1.88(m,1H),1.80–1.76(m,1H),1.64–1.61(m,1H),1.53–1.50(m,1H),1.22(s,1H).

[0156] Example 8 Preparation of (S)-5-(1H-pyrrolo[2,3-b]pyridin-3-yl)-7-((6,6-dimethyl)piperidine-3-amino)pyrazolo[1,5-a]pyrimidine-3-carboxylonitrile (8)

[0157]

[0158] Step 1: Preparation of (S)-3-tert-butoxycarbonylamino-(6,6-dimethyl)piperidine (8b)

[0159] Compound 8a (1 g, 4.7 mmol) was dissolved in tetrahydrofuran (20 mL), followed by the addition of zirconium chloride (3.3 g, 14.1 mmol). The mixture was stirred in an ice bath at -30°C for 0.5 hours under nitrogen protection. Then, methylmagnesium bromide (3.4 g, 28.2 mmol) was slowly added, and the mixture was stirred in an ice bath for 1 hour before being allowed to return to room temperature and stirred overnight. The reaction mixture was quenched with saturated ammonium chloride solution, extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer chromatography using silica gel to give 429 mg of the title compound (yield: 40%).

[0160] Step 2: Preparation of (S)-3-amino-(6,6-dimethyl)piperidine (8c)

[0161] Compound 8b (429 mg, 1.88 mmol) was dissolved in dichloromethane (4 mL), and then hydrochloric acid-ethyl acetate solution (4 mL, 4 M) was added. The reaction system was stirred at room temperature for 2 hours. The reaction system was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to give 169 mg of the title compound, yield: 70%.

[0162] Step 3: Preparation of (S)-3-(5-chloro-3-cyanopyrazolo[1,5-a]pyrimidin-7-yl)amino-(6,6-dimethyl)piperidine (8d)

[0163] Compound 8c (169 mg, 1.32 mmol) and compound 1f (308 mg, 1.45 mmol) were dissolved in acetonitrile (13 mL), and then triethylamine (267 mg, 2.64 mmol) was added. The reaction mixture was stirred at 50 °C for 4 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 165 mg of the title compound, yield: 41%.

[0164] Step 4: Preparation of (S)-5-(1H-pyrrolo[2,3-b]pyridin-3-yl)-7-((6,6-dimethyl)piperidin-3-amino)pyrazolo[1,5-a]pyrimidine-3-carboxylonitrile (8)

[0165] Compound 8d (165 mg, 0.54 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)-1H-pyrrolo[2,3-b]pyridine (195 mg, 0.80 mmol), Pd(dppf)Cl2 (38 mg, 0.054 mmol), and potassium carbonate (146 mg, 1.06 mmol) were dissolved in 1,4-dioxane / water (8 mL / 0.5 mL). The reaction mixture was heated to 110 °C and stirred overnight under nitrogen protection. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 28 mg of the title compound (yield: 14%).

[0166] LC-MS(ESI)m / z(M+H) + 387.2

[0167] 1H NMR (400MHz, DMSO-d6) δ12.38 (s, 1H), 8.92 (dd, J = 7.9, 1.7Hz, 1H), 8.66 (s, 1H), 8.58(s,1H),8.32(dd,J=4.7,1.7Hz,1H),7.86(s,1H),7.28(dd,J=8.0,4.6Hz,1H ),6.97(s,1H),3.84(s,1H),2.97–2.93(m,1H),2.87–2.82(m,1H),2.00–1.89(m, 1H),1.84–1.79(m,1H),1.58–1.52(m,1H),1.47–1.41(m,1H),1.15–1.09(m,6H).

[0168] Example 9 (S)-7-(dimethyloxophosphoryl)-3-(3-ethyl-7-(piperidin-3-amino)pyrazolo[1,5-a]pyrimidin-5-yl)-1H-indole-6-carboxynitrile

[0169] Compound 9 was prepared according to the preparation method in Example 4.

[0170]

[0171] Example 10 Preparation of (S)-5-(6-cyano-7-(dimethylphosphoryl)-1H-indol-3-yl)-7-(6,6-dimethylpiperidin-3-yl)amino)pyrazole[1,5-a]pyrimidine-3-carboxynitrile (10)

[0172]

[0173] Compounds 8d (120 mg, 0.39 mmol), 5c (142 mg, 0.41 mmol), Pd(dppf)Cl2 (27.4 mg, 0.039 mmol), and potassium carbonate (108 mg, 0.78 mmol) were dissolved in 1,4-dioxane / water (5 mL / 1 mL). The reaction mixture was heated to 110 °C and stirred overnight under nitrogen protection. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 24 mg of the title compound (yield: 12.7%).

[0174] LC-MS(ESI)m / z(M+H) + : 487.2

[0175] Example 11 Preparation of (S)-5-(6-cyano-1H-indol-3-yl)-7-((6,6-dimethyl)piperidine-3-amino)pyrazolo[1,5-a]pyrimidine-3-carboxynitrile (11)

[0176]

[0177] Step 1: Preparation of (S)-3-(7-((6,6-dimethyl)piperidin-3-yl)amino)-3-cyanopyrazolo[1,5-a]pyrimidin-5-yl)-6-cyano-1H-indole-1-carboxylic acid tert-butyl ester (11a)

[0178] Compound 8d (165 mg, 0.54 mmol), 6-cyano-3-(4,4,5,5-tetramethyl-1,3,2-dioxorane-2-yl)-1H-indole (295 mg, 0.80 mmol), Pd(dppf)Cl2 (38 mg, 0.054 mmol), and potassium carbonate (146 mg, 1.06 mmol) were dissolved in 1,4-dioxane / water (8 mL / 0.5 mL). Under nitrogen protection, the reaction mixture was heated to 110 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 55 mg of the title compound (yield: 20%).

[0179] Step 2: Preparation of (S)-5-(6-cyano-1H-indol-3-yl)-7-((6,6-dimethyl)piperidine-3-amino)pyrazolo[1,5-a]pyrimidine-3-carboxylonitrile (11)

[0180] Compound 11a (55 mg, 108 μmol) was dissolved in dichloromethane (4 mL), and then hydrochloric acid-ethyl acetate solution (2 mL, 4 M) was added. The reaction system was stirred at room temperature for 2 hours. The reaction system was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 27 mg of the title compound, yield: 61%.

[0181] LC-MS(ESI)m / z(M+H) + : 411.2

[0182] 1H NMR (400MHz, DMSO-d6) δ12.12(s,1H),9.19(s,1H),8.68(s,1H),8.59(d,J=3.6Hz ,1H),7.87(d,J=8.2Hz,1H),7.62(dd,J=8.1,1.5Hz,1H),7.02(d,J=3.6Hz,1H),6 .85(s,1H),3.93(s,1H),3.02–2.94(m,1H),2.75(t,J=9.8Hz,1H),1.90–1.80(m, 1H),1.77–1.64(m,1H),1.62–1.51(m,1H),1.47–1.35(m,1H),1.16–1.08(m,6H).

[0183] Example 12 Preparation of (S)-N-(3-(3-cyano-7-(piperidin-3-amino)pyrazolo[1,5-a]pyrimidin-5-yl)phenyl)acetamide (12)

[0184]

[0185] Step 1: Preparation of (S)-3-((5-(3-acetamidophenyl)-3-cyanopyrazolo[1,5-a]pyrimidin-7-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (12a)

[0186] 1 g (100 mg, 0.27 mmol) of the compound, N-(3-(4,4,5,5-tetramethyl-1,3,2-dioxoboron-2-yl)phenyl)acetamide (75 mg, 0.28 mmol), Pd(dppf)Cl2 (19 mg, 0.027 mmol), and potassium carbonate (74.5 mg, 0.54 mmol) were dissolved in 1,4-dioxane / water (5 mL / 1 mL). Under nitrogen protection, the reaction mixture was heated to 110 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 67 mg of the title compound, yield: 52.3%.

[0187] Step 2: Preparation of (S)-N-(3-(3-cyano-7-(piperidin-3-amino)pyrazolo[1,5-a]pyrimidin-5-yl)phenyl)acetamide (12)

[0188] Compound 12a (67 mg, 0.14 mmol) was dissolved in ethyl acetate (4 mL), and then hydrochloric acid-ethyl acetate solution (2 mL, 4 M) was added. The reaction system was stirred at room temperature for 2 hours. The reaction system was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 24 mg of the title compound, yield: 46.1%.

[0189] LC-MS(ESI)m / z(M+H) + 376.2

[0190] 1 H NMR (400MHz, DMSO-d6) δ10.21(s,1H),8.67(s,1H),8.25(t,J=1.9Hz,1H),8.20(s,1H),7.94(d d,J=8.1,2.1Hz,1H),7.89-7.83(m,1H),7.46(t,J=8.0Hz,1H),6.96(s,1H),4.03-3.92(m,1H), 3.05(dd,J=11.9,3.6Hz,1H),2.84-2.78(m,1H),2.74-2.66(m,1H),2.59-2.52(m,1H),2.08(s ,3H),1.95-1.86(m,1H),1.84-1.75(m,1H),1.70-1.62(m,1H),1.56-1.50(m,1H),1.22(s,1H).

[0191] Example 13 Preparation of (S)-7-(piperidin-3-ylamino)-5-(1H-pyrazol-4-yl)pyrazol[1,5-a]pyrimidine-3-carboxynitrile (13)

[0192]

[0193] Step 1: Preparation of (S)-3-((3-cyano-5-(1H-pyrazol-4-yl)pyrazol[1,5-a]pyrimidin-7-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (12a)

[0194] 1 g (100 mg, 0.27 mmol) of the compound, 54.6 mg (0.28 mmol) of 4-(4,4,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)-1H-pyrazole, 19 mg (0.027 mmol) of Pd(dppf)Cl2, and 74.5 mg (0.54 mmol) of potassium carbonate were dissolved in 1,4-dioxane / water (5 mL / 1 mL). The reaction mixture was heated to 110 °C and stirred overnight under nitrogen protection. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to give 51 mg of the title compound, yield: 46.3%.

[0195] Step 2: Preparation of (S)-7-(piperidin-3-ylamino)-5-(1H-pyrazol-4-yl)pyrazol[1,5-a]pyrimidine-3-carboxynitrile (13)

[0196] Compound 13a (51 mg, 0.12 mmol) was dissolved in ethyl acetate (4 mL), and then hydrochloric acid-ethyl acetate solution (2 mL, 4 M) was added. The reaction system was stirred at room temperature for 2 hours. The reaction system was concentrated under reduced pressure, diluted with water, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer silica gel chromatography to obtain 22 mg of the title compound, yield: 59.4%.

[0197] LC-MS(ESI)m / z(M+H) + 309.2

[0198] 1 H NMR (400MHz, DMSO-d6) δ13.30(s,1H),8.58(s,1H),8.38(s,2H),7.96(s,1H),6.85(s,1H),3.96-3.92(m,1H),3.06(dd,J=11.9,3.5Hz,1H),2.88- 2.84(m,1H),2.73-2.70(m,1H),2.62-2.54(m,1H),1.92-1.88(m,1H),1. 79-1.75(m,1H),1.72-1.64(m,1H),1.58-1.53(m,1H),1.23-1.21(m,1H).

[0199] Example 14 (S)-5-(6-cyano-7-methanesulfonyl-1H-indol-3-yl)-7-((6,6-dimethyl)piperidine-3-amino)pyrazolo[1,5-a]pyrimidine-3-carboxynitrile

[0200] Compound 14 was prepared according to the preparation method of Example 8.

[0201]

[0202] Example 15 (S)-7-((6,6-dimethyl)piperidine-3-amino)-5-(7-(1,1-dioxothiomorpholinyl)-1H-indol-3-yl)-pyrazolo[1,5-a]pyrimidine-3-carboxynitrile

[0203] Compound 15 was prepared according to the preparation method of Example 8.

[0204]

[0205] Example 16 (S)-5-(6-(3,5-dimethylisoxazol-4-yl)-pyrrolo[2,3-b]pyridin-3-yl)-7-((6,6-dimethyl)piperidine-3-amino)-pyrazolo[1,5-a]pyrimidine-3-carboxynitrile

[0206] Compound 16 was prepared according to the preparation method of Example 8.

[0207]

[0208] Biological evaluation

[0209] Experimental Example 1: CDK7 Enzyme Activity Assay

[0210] 1. Experimental Objective

[0211] This experiment used a three-step method to detect the inhibitory activity of compounds on the CDK7 enzyme by measuring the amount of ADP generated in the kinase reaction. The first step involved incubating CDK7 / CyclinH1 / MNAT1, different concentrations of the compound, substrate (MBP), and ATP at room temperature for a fixed time. The second step involved adding ADPglo assay reagent to terminate the enzymatic reaction, followed by further incubation at room temperature for a fixed time. The third step involved adding Kinase Detection Reagent, incubating at room temperature for a fixed time, and then reading the values. Continuous readings were performed using the Lumi module of a BMG microplate reader to evaluate the effect of the test compounds on CDK7 enzyme activity. The IC50 of the test compounds on CDK7 enzyme was calculated using the inhibition rate. 50 value.

[0212] 2. Test materials

[0213] 2.1 Compounds:

[0214] Test drug: Compound 3 was prepared by the synthetic method of Example 3;

[0215] Test drug: Compound 4 was prepared by the synthetic method of Example 4;

[0216] Test drug: Compound 5 was prepared by the synthetic method of Example 5;

[0217] Test drug: Compound 6 was prepared by the synthetic method of Example 6;

[0218] Test drug: Compound 7 was prepared by the synthetic method of Example 7;

[0219] Test drug: Compound 8 was prepared by the synthetic method of Example 8;

[0220] Test drug: Compound 10 was prepared by the synthetic method of Example 10;

[0221] Test drug: Compound 11 was prepared by the synthetic method of Example 11;

[0222] 2.2 Test Reagents and Instruments

[0223] CDK7 / CyclinH1 / MNAT1, Promega;

[0224] ADPglo, Promega;

[0225] DMSO, Sigma;

[0226] OptiPlate-384, Thermo;

[0227] Microplate reader (BMG), POLARstar;

[0228] Pipettes, Eppendorf;

[0229] 3. Test methods

[0230] 3.1 Compound Preparation

[0231] Preparation of 10mM compound stock solution: Dissolve the compound powder in 100% DMSO to prepare 10mM compound stock solutions.

[0232] 3.2 Enzyme Reaction Process

[0233] (1) Prepare 1× buffer solution.

[0234] (2) Compound concentration preparation: Compound IC 50 The test started at a final concentration of 10 μM, diluted 5-fold, resulting in 6 concentrations, with single / duplicate assays set up for each concentration. 1 μL of the compound was added to each well and diluted 5 times the corresponding final concentration in a 384-well plate. For the negative control, no compound or CDK7 / CyclinH1 / MNAT1 was added, and the remaining volume was brought to 1× reaction solution. For the positive control, no compound was added, and the remaining volume was brought to 1× reaction solution.

[0235] (3) Prepare CDK7 / CyclinH1 / MNAT1 enzyme (15ng / well) solution with 1× reaction solution, and add 2μL to each well.

[0236] (4) Prepare a mixed solution of substrate and ATP (0.33 ng / well) using 1× reaction solution, and add 2 μL to each well.

[0237] (5) Start the reaction and incubate at room temperature for 60 minutes.

[0238] (6) Add 5 μL of ADPglo reagent to each well and incubate at room temperature for 50 min.

[0239] (7) Add 10 μL of detection solution to each well and incubate at room temperature for 30 min.

[0240] (8) Read the signal value using the Lumi module of the BMG microplate reader, and read continuously (1 min / time) for a total of 30 times.

[0241] 3.3 Data Analysis

[0242] Calculation formula

[0243]

[0244] Plotting the logarithm of concentration on the X-axis and the percentage inhibition rate on the Y-axis, dose-response curves were fitted using the log(inhibitor) vs. response-variable slop(four parameters) formula in Graphpad Prism 9 software to derive the IC50 of each compound on enzyme activity. 50 value.

[0245] 4. Test Results

[0246] The inhibitory activity of the compounds in this application against CDK7 enzyme is shown in Table 1.

[0247] Table 1. Inhibitory activity of the compounds in this application against CDK7 enzyme.

[0248] Compound numbering <![CDATA[IC 50 (nM)]]> Compound numbering <![CDATA[IC 50 (nM)]]> 3 1.60 4 4.80 5 5.80 6 0.47 7 0.65 8 0.72 10 0.08 11 0.81

[0249] As can be seen from the test data of CDK7 enzyme inhibition of the compounds in Table 1, the compounds of this application have obvious CDK7 enzyme inhibition activity.

[0250] It will be apparent to those skilled in the art that various modifications and variations can be made to the compounds and their preparation methods without departing from the spirit or scope of this application. Therefore, the scope of protection of this application covers various modifications and variations made to this application, as long as the modifications or variations are within the scope covered by the claims and their equivalent embodiments.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, in, Ring A is selected from 5-10 membered heterocyclic aryl groups, wherein the heterocyclic aryl group contains at least one nitrogen atom as a heteroatom; The compound with structure I is a compound with structure II. Y 1 Y 2 Y 3 Y 4 Y 5 Selected independently from: C, N, CR 1 or NR 6 ; R 1 Selected from: hydrogen, cyano, C1-C6 alkyl, C3-C8 cycloalkyloxy; R 2 Selected from: hydrogen, halogen, cyano, or C1-C6 alkyl; R 3 Selected from: hydrogen, halogen, cyano, or C1-C6 alkyl; R 4 R 5 Each is independently selected from: hydrogen or C1-C6 alkyl; R 6 Selected from: hydrogen; m is selected from: 1, 2, or 3; n is selected from: 1 or 2.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The compound is selected from: Y 3 For NR 6 ; and / or, R 1 Selected from: hydrogen, cyano, C1-C6 alkyl; and / or, R 2 Selected from: hydrogen, cyano, or C1-C6 alkyl; and / or, R 3 Selected from hydrogen, cyano, or C1-C6 alkyl; and / or, R 4 R 5 Each is independently selected from: hydrogen or C1-C6 alkyl; R 6 Selected from: hydrogen; m is selected from: 1 or 2; n is selected from: 1 or 2.

3. A compound or a pharmaceutically acceptable salt thereof, said compound being selected from:

4. A pharmaceutical composition, characterized in that, It comprises the compound of any one of claims 1-3 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

5. Use of the compound according to any one of claims 1-4 in the preparation of a medicament for the prevention or treatment of cyclin-dependent kinase 7 (CDK7) mediated diseases.

6. The use according to claim 5, characterized in that: The diseases mediated by the cyclin-dependent kinase 7 (CDK7) are selected from tumors or cancers.

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

8. A method for preparing the compound according to claim 1 or 2, comprising the following steps: Where LG and LG' represent leaving groups, and R 1 R 2 R 3 Ring A, ring B, m, and n are defined as described in claim 1 or 2, respectively.

9. The method for preparing the compound according to claim 8, characterized in that, The leaving group is selected from halogen atoms, methanesulfonyloxy groups, or p-toluenesulfonyloxy groups; (1) React compound I-1 with compound I-2 to obtain I-3; (2) The compound I-3 and the boric acid or borate ester of the compound I-4 are coupled together to obtain the compound with the structure shown in Formula I.

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