(1H-indazole-5-yl) amino-2-pyridine derivative and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a (1H-indazole-5-yl) amino-2-pyridine derivative and application thereof. The invention relates to a compound shown in a formula (I) or pharmaceutically acceptable salts thereof, and A, L, X, R1 and R2 are described in the specification. The compound disclosed by the invention has relatively strong ROCK2 inhibitory activity in vitro, and can be used for treating ROCK2 related diseases, such as tumors or fibrosis and the like. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to a (1H-indazole-5-yl)amino-2-pyridine derivative and application thereof. Background Art

[0002] ROCK (Rho-associated protein kinase), also known as Rho kinase, is a serine / threonine protein kinase and the downstream effector of the small molecule GTPase Rho, whose function has been most thoroughly studied. ROCK is widely involved in a series of biological processes such as cell mitosis, cytoskeleton remodeling, smooth muscle cell contraction, nerve regeneration, tumor cell infiltration, and regulation of cell apoptosis by regulating the reorganization of the cell actin skeleton. A large number of studies have shown that ROCK is closely related to the occurrence and development of a variety of diseases such as eye diseases, cardiovascular and cerebrovascular diseases, autoimmune diseases, tumors, and fibrosis.

[0003] At present, a variety of ROCK inhibitors have been developed at home and abroad. According to the different structures of the hinge binding region of the kinase, the existing ROCK inhibitors can be roughly divided into isoquinoline, 4-aminopyridine, biaryl and indazole. The structural characteristics of isoquinoline ROCK inhibitors are that they have an isoquinoline structure and a homopiperazine ring, and the two are connected by a sulfonyl group. The representative drugs are Fasudil and Ripasudil (WO2020177291, US2008064681), which are used to treat amyotrophic lateral sclerosis and glaucoma, respectively. The 4-aminopyridine compound has a benzene ring or cyclohexane structure in the center of the molecule, and has a side chain at the 4th position of the cyclohexane. The representative drug is Netarsudil (WO2017086941), which is used to treat open-angle glaucoma. The structural characteristics of biaryl ROCK inhibitors are generally that they have a nitrogen-containing small molecule heterocycle at the end, which is then connected to an aromatic ring that acts as a hydrophobic ring through an amide bond or a urea structure to stabilize the conformation of the compound. Representative drugs include the technology disclosed in invention patent WO2011130740.

[0004] Indazole compounds are a novel class of ROCK inhibitors. Due to their high selectivity for ROCK2 subtypes, they have attracted widespread attention from researchers in recent years. The representative drug Belumosudil was approved for the treatment of chronic graft-versus-host disease in 2021. It also has good effects on the treatment of fibrotic diseases. However, due to its lack of outstanding inhibitory efficacy, it has not made significant progress in the treatment of other diseases. . The technologies disclosed in invention patents US201314136359, CA2922312, WO2024118556, CN116891460, CN114504574, etc. are all ROCK inhibitors containing indazole structures. In addition to indazole as a protein hinge binding region, their structural features are different aromatic groups at the end of the molecule combined with a protein hydrophobic pocket, and the middle linker is an amide, urea, aromatic ring or a simple aliphatic ring, but most of them lack targeted efficacy against tumors or fibrosis. So far, no ROCK inhibitors for the treatment of tumors or fibrosis have been approved. Summary of the invention

[0005] In view of the problems existing in the background technology, the present invention provides a (1H-indazol-5-yl)amino-2-pyridine derivative and application thereof.

[0006] To achieve the above purpose, the present invention adopts the following technical solution:

[0007] A (1H-indazol-5-yl)amino-2-pyridine derivative, wherein the derivative is a compound represented by the general formula (I) and a pharmaceutically acceptable salt thereof,

[0008]

[0009] Where:

[0010] L is

[0011] R1 is unsubstituted or substituted by at least one identical or different R5 (C1-C6) alkyl, (C1-C6) alkoxy, (C3-C8) cycloalkyl, (C 2- C6)alkenyl, (C2-C6)alkynyl, or (CH2) p NR3R4;

[0012] R5 is halogen, hydroxy, amino, cyano or carboxyl;

[0013] p is independently 0, 1, 2 or 3;

[0014] R3 and R4 are the same or different and are independently selected from hydrogen, (C1-C6) alkyl and (C3-C8) cycloalkyl which are unsubstituted or substituted by at least one identical or different R6; or R3 and R4 together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclic group containing at least one heteroatom, wherein the heterocyclic group is optionally substituted by 0-3 independent hydroxyl groups, (C1-C6) alkyl, (C1-C6) alkoxy or (C1-C6) alkoxycarbonyl which are unsubstituted or substituted by at least one identical or different R6;

[0015] R6 is halogen, amino, hydroxy, cyano or (C3-C6) cycloalkyl;

[0016] A is a 4-7 membered heterocyclic group containing 1-3 heteroatoms selected from N, O and S;

[0017] X is selected from

[0018] R2 is selected from (C1-C6)alkyl, (C3-C6)cycloalkyl, 4-6 membered heterocyclyl, aryl, which is unsubstituted or substituted by at least one identical or different R5;

[0019] R7 is halogen, hydroxy, nitro, cyano, (C1-C6) alkyl, (C1-C6) alkoxy or (C1-C6) alkylamino.

[0020] Preferably, the derivative is a compound represented by the general formula (I) and a pharmaceutically acceptable salt thereof, wherein:

[0021] L is

[0022] R1 is unsubstituted or substituted by at least one identical or different R5 (C1-C6) alkyl, (C3-C8) cycloalkyl, (C2-C6) alkenyl or (CH2) p NR3R4;

[0023] R5 is halogen, hydroxy, amino, cyano or carboxyl;

[0024] wherein p is independently 0, 1, 2 or 3;

[0025] R3 and R4 are the same or different and are independently selected from hydrogen, (C1-C6)alkyl and (C3-C6)cycloalkyl which are unsubstituted or substituted by at least one identical or different R6; or R3 and R4 together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclic group containing at least one heteroatom, wherein the heterocyclic group is optionally substituted by 0-3 independent hydroxyl groups, (C1-C6)alkyl groups, (C1-C6)alkyl groups which are unsubstituted or substituted by at least one identical or different R6, (C1-C6)alkoxy groups or (C1-C6)alkoxycarbonyl groups;

[0026] R6 is halogen, amino, hydroxy, cyano or (C3-C6) cycloalkyl;

[0027] A is a 4-7 membered heterocyclic group containing 1-3 heteroatoms selected from N or O;

[0028] X is selected from One of;

[0029] R2 is selected from (C1-C6)alkyl, (C3-C6)cycloalkyl, 4-6 membered heterocyclyl, aryl, which is unsubstituted or substituted by at least one identical or different R5;

[0030] R7 is halogen, hydroxy, nitro, cyano, (C1-C6) alkyl, (C1-C6) alkoxy or (C1-C6) alkylamino.

[0031] More preferably, the derivative is a compound represented by the general formula (I) and a pharmaceutically acceptable salt thereof, wherein:

[0032] L is

[0033] R1 is unsubstituted or substituted by at least one identical or different R5 (C1-C4) alkyl, (C3-C6) cycloalkyl, (C2-C4) alkenyl or (CH2) p NR3R4;

[0034] R5 is halogen, hydroxy, amino, cyano or carboxyl;

[0035] Where p is independently 1, 2 or 3

[0036] R3 and R4 are the same or different and are independently selected from hydrogen, (C1-C4) alkyl and (C3-C6) cycloalkyl which are unsubstituted or substituted by at least one identical or different R6; or R3 and R4 together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclic group containing at least one heteroatom, wherein the heterocyclic group is optionally substituted by 0-1 independent hydroxyl groups, (C1-C3) alkyl groups, (C1-C3) alkyl groups which are unsubstituted or substituted by at least one identical or different R6, or (C1-C3) alkoxycarbonyl groups;

[0037] R6 is amino, hydroxy or (C3-C6) cycloalkyl;

[0038] A is a 5-6 membered heterocyclic group containing 1-2 heteroatoms selected from N or O;

[0039] X is selected from One of;

[0040] R2 is selected from (C1-C6) alkyl, (C3-C6) cycloalkyl, 5-6 membered heterocyclyl, aryl, which is unsubstituted or substituted by at least one identical or different R5;

[0041] R7 is halogen, hydroxy, nitro, cyano, (C1-C4) alkyl or (C1-C4) alkylamino.

[0042] Further preferably, the derivative is a compound represented by the general formula (I) and a pharmaceutically acceptable salt thereof, wherein:

[0043] L is

[0044] R1 is (C1-C4)alkyl, (C2-C4)alkenyl or CH2NR3R4;

[0045] R3 and R4 are the same or different and are independently selected from hydrogen, (C1-C4) alkyl and (C3-C6) cycloalkyl which are unsubstituted or substituted by at least one identical or different R6; or R3 and R4 together with the nitrogen atom to which they are attached form a 5-6 membered heterocyclic group containing at least one heteroatom, wherein the heterocyclic group is optionally substituted by 0-1 independent hydroxyl, (C1-C3) alkyl, (C1-C3) alkyl which is unsubstituted or substituted by at least one identical or different R6, or methoxycarbonyl;

[0046] R6 is hydroxy or cyclopropyl;

[0047] A is

[0048] X is selected from One of;

[0049] R2 is selected from (C1-C6) alkyl, (C3-C6) cycloalkyl, 4-piperidinyl, aryl which is unsubstituted or substituted by at least one identical or different R7;

[0050] R7 is halogen, (C1-C4) alkyl or (C1-C4) alkylamino.

[0051] Still more preferably, the derivative is a compound represented by the general formula (I) and a pharmaceutically acceptable salt thereof, wherein:

[0052] L is

[0053] R1 is methyl, tert-butyl, vinyl,

[0054]

[0055] X is selected from One of;

[0056] A is

[0057] R2 is

[0058] Most preferably, the derivative is the following compound and a pharmaceutically acceptable salt thereof,

[0059] N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(pyrrolidin-1-yl)acetamide;

[0060] N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-morpholinoacetamide;

[0061] N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(piperidin-1-yl)acetamide;

[0062] N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(cyclopentylamino)acetamide;

[0063] N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(4-methylpiperazin-1-yl)acetamide;

[0064] N-{6-[(1H-indazol-5-yl)amino]-2-{4-[4-(dimethylamino)benzoyl]piperazin-1-yl}pyridin-3-yl}-2-(pyrrolidin-1-yl)acetamide;

[0065] N-{6-[(1H-indazol-5-yl)amino]-2-{4-[4-(dimethylamino)benzoyl]piperazin-1-yl}pyridin-3-yl}-2-(dimethylamino)acetamide;

[0066] N-{6-[(1H-indazol-5-yl)amino]-2-{4-[4-(dimethylamino)benzoyl]piperazin-1-yl}pyridin-3-yl}-2-(cyclopropylamino)acetamide;

[0067] N-{6-[(1H-indazol-5-yl)amino]-2-{4-[4-(dimethylamino)benzoyl]piperazin-1-yl}pyridin-3-yl}-2-[(2-hydroxyethyl)amino]acetamide;

[0068] N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-methylbenzoyl)piperazin-1-yl]pyridin-3-yl}-2-[(2-hydroxyethyl)amino]acetamide;

[0069] N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-methylbenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(4-hydroxypiperidin-1-yl)acetamide;

[0070] N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-methylbenzoyl)piperazin-1-yl]pyridin-3-yl}-2-[4-(2-hydroxyethyl)piperazin-1-yl]acetamide;

[0071] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-(methylsulfonamido)pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate;

[0072] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl{4-(dimethylamino)phenyl}carbamate;

[0073] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(dimethylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate;

[0074] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-methylpiperazin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate;

[0075] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[(cyclopropylmethyl)amino]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate;

[0076] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[(2-hydroxyethyl)amino]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate;

[0077] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-acrylamidopyridin-2-yl}pyrrolidin-2-yl}p-tolylcarbamate;

[0078] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-methylpiperazin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}p-tolylcarbamate;

[0079] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopentylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}p-tolylcarbamate;

[0080] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-hydroxypiperidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl p-tolylcarbamate;

[0081] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-ethylpiperazin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl p-tolylcarbamate;

[0082] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[4-(2-hydroxyethyl)piperazin-1-yl]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}p-tolylcarbamate;

[0083] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl (4-fluorophenyl)carbamate;

[0084] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-pivalamidopyridin-2-yl}pyrrolidin-2-yl}methyl (4-fluorophenyl)carbamate;

[0085] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-hydroxypiperidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl (4-fluorophenyl)carbamate;

[0086] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[(cyclopropylmethyl)amino]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}methyl (4-fluorophenyl)carbamate;

[0087] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylpiperidin-4-ylcarbamate;

[0088] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-(2-thiomorpholinoacetylamino)pyridin-2-yl}pyrrolidin-2-yl}methylpiperidin-4-ylcarbamate;

[0089] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-methylpiperidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylpiperidin-4-ylcarbamate;

[0090] (S)-1-{2-{{6-[(1H-indazol-5-yl)amino]-2-{2-[(piperidin-4-ylcarbamoyl)oxy]methyl}pyrrolidin-1-yl}pyridin-3-yl}amino-2-oxoethyl}piperidine-4-carboxylic acid methyl ester;

[0091] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl isopropylcarbamate;

[0092] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopropylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl isopropylcarbamate;

[0093] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopentylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl isopropylcarbamate;

[0094] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylcyclopropylcarbamate;

[0095] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopropylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylcyclopropylcarbamate;

[0096] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylcyclohexylcarbamate;

[0097] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-(2-morpholinoacetylamino)pyridin-2-yl}pyrrolidin-2-yl}methylcyclohexylcarbamate;

[0098] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopropylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylcyclohexylcarbamate;

[0099] (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[(cyclopropylmethyl)amino]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}methylcyclohexylcarbamate;

[0100] (S)-N-{{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-methylpiperazin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl}-4-(dimethylamino)benzamide.

[0101] The salt of the compound represented by the general formula (I) is a pharmaceutically acceptable salt obtained by reacting the compound with an acid, and the corresponding acid is hydrochloric acid, hydrobromic acid, sulfuric acid, lactic acid, citric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, tartaric acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid.

[0102] An application of the (1H-indazol-5-yl)amino-2-pyridine derivative, and an application of the compound represented by the general formula (I) and a pharmaceutically acceptable salt thereof as a ROCK2 inhibitor in the preparation of a drug for treating tumors or fibrotic diseases.

[0103] The tumor is selected from lung cancer, breast cancer, gastric cancer, colon cancer, hepatocellular carcinoma, oral cancer, kidney cancer, bladder cancer, ovarian cancer, cervical cancer, neuroblastoma, osteosarcoma, and fibrosarcoma; the fibrosis is selected from pulmonary fibrosis, renal fibrosis, liver fibrosis, and myocardial fibrosis.

[0104] Beneficial effects of the present invention

[0105] The present invention provides a (1H-indazol-5-yl)amino-2-pyridine derivative as a ROCK2 inhibitor, which fills the research gap; the (1H-indazol-5-yl)amino-2-pyridine derivative not only has a novel structure type, but also exhibits high inhibitory activity on ROCK2. In an in vitro cell experiment, the (1H-indazol-5-yl)amino-2-pyridine derivative can effectively inhibit the proliferation of MDA-MB-231 breast cancer cells and TGF-β-induced NIH-3T3 mouse embryonic fibroblasts and inhibit the migration of MDA-MB-231 cells. At the same time, in a bleomycin-induced C57BL / 6 mouse pulmonary fibrosis model, the (1H-indazol-5-yl)amino-2-pyridine derivative can effectively improve the degree of inflammation of mouse lung tissue, and has the therapeutic potential for tumors and fibrotic diseases. The present invention also provides a preparation method of a (1H-indazol-5-yl)amino-2-pyridine derivative, which is simple in preparation, easy to purify, and the required raw materials are easily available, and have good feasibility. BRIEF DESCRIPTION OF THE DRAWINGS

[0106] Figure 1 These are the results of the anti-MDA-MB-231 cell migration experiment with compound 21.

[0107] Figure 2 The results of HE staining of mouse lung tissue. DETAILED DESCRIPTION

[0108] The present invention is further described below in conjunction with specific implementation cases, but the present invention is not limited to these embodiments.

[0109] The present invention combines indazole fragments and introduces the key pharmacophore aminopyridine structure in anti-tumor drugs as a linker. The (1H-indazole-5-yl)amino-2-pyridine derivatives formed are a new ROCK2 inhibitor. This type of compound not only has a novel structure type, but also has a high inhibitory effect on ROCK2 and exhibits diverse biological activities in vitro and in vivo. It is expected to improve the targeted therapeutic effect on tumors and fibrosis, and is a highly potential drug structure type. At present, no (1H-indazole-5-yl)amino-2-pyridine derivatives have been reported as ROCK2 inhibitors.

[0110] In the following examples, the H NMR spectra of the compounds were measured using Bruker ARX-400 / 600, and the mass spectra were measured using Agilent 110 LC / MSD; all reagents used were of analytical or chemical purity.

[0111] Examples 1-12

[0112]

[0113]

[0114] Embodiment 13

[0115]

[0116] Embodiment 14-41

[0117]

[0118]

[0119]

[0120] Embodiment 42

[0121]

[0122] Example 1: N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(pyrrolidin-1-yl)acetamide (1);

[0123] The route looks like this:

[0124]

[0125] Step A1: Synthesis of Intermediate A-I

[0126] 5-nitroindazole (5.0 g, 61.3 mmol) was dissolved in ethyl acetate (50 mL), p-toluenesulfonic acid monohydrate (2.3 g, 12.3 mmol) and 3,4-2H-pyran (15.5 g, 183.9 mmol) were added, and the mixture was stirred at room temperature for 2 h. Water (30 mL) was added to the reaction solution, and the organic phase was washed twice with water (30 mL), and the organic phase was evaporated to dryness. The crude product was recrystallized from methanol to obtain 12.5 g of a light yellow solid with a yield of 83.2%.

[0127] Step A2: Synthesis of intermediate A-II

[0128] Dissolve the intermediate A-Ⅰ (10.0 g, 40.5 mmol) in a mixed solution of methanol (50 mL) and tetrahydrofuran (50 mL), add wet palladium carbon (1.0 g, 5%), and stir at room temperature under hydrogen for 8 h. Filter the reaction solution, wash the filter cake with methanol (10 mL), and evaporate the filtrate to obtain 8.1 g of lavender solid, with a yield of 92.0%. Analytical data: ESI-MS [M+H] + (m / z): 218.3.

[0129] Step A3: Synthesis of intermediate A-III

[0130] Dissolve 2,6-dichloro-3-nitropyridine (10.0 g, 52.09 mmol) in tetrahydrofuran (150 mL), add potassium carbonate (14.4 g, 104.18 mmol), slowly add N-Boc-piperazine (8.8 g, 47.35 mmol) at low temperature, and stir at room temperature for 12 h. The reaction solution was filtered, the filtrate was evaporated to dryness, and the crude product was purified by column chromatography to obtain 13.6 g of yellow solid, with a yield of 76.3%. Analytical data: ESI-MS [M+Na] + (m / z):365.0.

[0131] Step A4: Synthesis of Intermediate A-IV

[0132] Dissolve intermediate A-Ⅲ (10.0 g, 29.2 mmol) in 4M hydrochloric acid / ethyl acetate solution (80 mL) and stir at room temperature for 3 h. Filter the reaction solution, wash the filter cake twice with ethyl acetate (20 mL), and dry to obtain 7.1 g of light yellow solid, with a yield of 78.6%. Analytical data: ESI-MS [M+H] + (m / z): 242.3.

[0133] Step A5: Synthesis of Intermediate A-V

[0134] Dissolve 4-fluorobenzoyl chloride (1.0 g, 6.4 mmol) and N,N-diisopropylethylamine (1.4 g, 10.8 mmol) in dichloromethane (15 mL). Add intermediate A-IV (1.5 g, 5.4 mmol) in batches under ice bath and stir at room temperature for 1 h. Add water (10 mL) to the reaction solution and stir. Separate the organic phase, wash the organic phase twice with saturated sodium bicarbonate solution (5 mL), dry over anhydrous sodium sulfate, and evaporate to dryness to obtain 2.15 g of yellow solid, with a yield of 99.1%. Analytical data: ESI-MS [MH] - (m / z): 363.2.

[0135] Step A6: Synthesis of Intermediate A-VI

[0136] Dissolve intermediate A-V (0.4 g, 1.1 mmol) and intermediate A-II (0.29 g, 1.3 mmol) in DMF (5 mL), add N,N-diisopropylethylamine (0.21 g, 1.65 mmol), and stir at 80°C for 12 h. Pour the reaction solution into water (200 mL) and stir, a large amount of yellow solid precipitates, filter, wash the filter cake with water (5 mL), dry, and purify the crude product by column chromatography to obtain 0.6 g of yellow solid, with a yield of 64.1%. Analytical data: ESI-MS [M+H] + (m / z):568.4.

[0137] Step A7: Synthesis of Intermediate A-VII

[0138] Dissolve intermediate A-VI (0.3 g, 0.55 mmol) in methanol (5 mL), add wet palladium carbon (0.03 g, 5%), and stir at room temperature under hydrogen for 8 h. Filter the reaction solution, wash the filter cake with methanol (3 mL), and evaporate the filtrate to dryness to obtain 0.27 g of green solid, with a yield of 98%. Analytical data: ESI-MS [M+H] + (m / z):516.4.

[0139] Step A8: Synthesis of Intermediate A-VIII

[0140] Dissolve the intermediate A-VII (0.3 g, 0.58 mmol) in methanol, add N,N-diisopropylethylamine (0.15 g, 0.87 mmol), add chloroacetyl chloride (0.09 g, 0.87 mmol) dropwise at low temperature, and stir at room temperature for 30 min. After the reaction solution is evaporated to dryness, add water (15 mL) and stir, filter, wash the filter cake with water (5 mL), dry, and purify the crude product by column chromatography to obtain 0.2 g of light yellow solid, with a yield of 58%. Analytical data: ESI-MS [M+H] + (m / z):592.4.

[0141] Step A9: Synthesis of Example 1

[0142] The intermediate A-Ⅷ (0.11 g, 0.25 mmol) was dissolved in tetrahydrofuran (2 mL), potassium carbonate (0.06 g, 0.50 mmol) and tetrahydropyrrole (0.02 g, 0.38 mmol) were added, and the mixture was stirred at 45 °C for 2 h. After the reaction solution was evaporated to dryness, water (5 mL) was added and stirred, and an off-white solid was precipitated and filtered. After the filter cake was dried, it was dissolved in dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added dropwise, and the mixture was stirred at room temperature for 2 h. The reaction solution was evaporated to dryness, water (5 mL) was added and stirred, and the solution was adjusted to pH 8 with a saturated sodium bicarbonate solution, extracted with dichloromethane (5 mL), and the organic phase was dried with anhydrous sodium sulfate and evaporated to dryness. The crude product was purified by column chromatography to obtain 79 mg of a white solid with a yield of 59.0%. Analytical data: ESI-MS [M+H] + (m / z):543.2. 1H NMR (400MHz, DMSO-d6) δ12.84(s,1H),9.28(s,1H),8.85(s,1H),8.24(d,J=8.7Hz,1H),8.11(s,1H),7.93(s,1H),7.58-7.50(m,2H),7.45(d,J =8.8Hz,2H),7.31(t,J=8.9Hz,2H),6.54(d,J=8.7Hz,1H),3.82(m,4H),3.25(s,2H),3.09(s,4H),2.66(d,J=6.0Hz,4H),1.80(q,J=3.2Hz,4H). 13 C NMR (101MHz, DMSO) δ168.82,168.60,164.27,161.82,151.69,150.87,136.37,135.42,133.32,132.78,132.74,131 .28,130.10,130.02,123.63,121.11,117.41,116.01,115.80,110.49,107.36,105.02,59.89,53.49,49.22,23.60.

[0143] Example 2: N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-morpholinoacetamide (2);

[0144] According to the method of step A9 in Example 1, intermediate A-VIII was used as a raw material, and after a nucleophilic substitution reaction with morpholine, deprotection was performed under acidic conditions to obtain Example 2 with a yield of 63.2%. Analytical data: ESI-MS [M+H] + (m / z):559.5.

[0145] Example 3: N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(piperidin-1-yl)acetamide (3);

[0146] According to the method of step A9 in Example 1, intermediate A-VIII was used as a raw material, and after a nucleophilic substitution reaction with piperidine, deprotection was performed under acidic conditions to obtain Example 3 with a yield of 54.3%. Analytical data: ESI-MS [M+H] + (m / z):557.6.

[0147] Example 4: N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(cyclopentylamino)acetamide (4);

[0148] According to the method of step A9 in Example 1, intermediate A-VIII was used as a raw material, and after a nucleophilic substitution reaction with cyclopentylamine, deprotection was performed under acidic conditions to obtain Example 4 with a yield of 43.4%. Analytical data: ESI-MS [M+H] + (m / z):557.5.

[0149] Example 5: N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(4-methylpiperazin-1-yl)acetamide (5);

[0150] According to the method of step A9 in Example 1, intermediate A-VIII was used as a raw material, and after a nucleophilic substitution reaction with 4-methylpiperazine, deprotection was performed under acidic conditions to obtain Example 5 with a yield of 55.2%. Analytical data: ESI-MS [M+H] + (m / z):572.3. 1 H NMR (400MHz, DMSO-d6) δ12.84(s,1H),9.20(s,1H),8.84(s,1H),8.23(d,J=8.6Hz,1H),8.10(s,1H),7.93(s,1H),7.54(dd,J=8.4,5.4Hz,2H) ,7.44(d,J=2.8Hz,2H),7.31(t,J=8.7Hz,2H),6.54(d,J=8.7Hz,1H),3.95-3.55(m,5H),3.11(s,6H),2.55(s,4H),2.40(s,4H),2.19(s,3H). 13 C NMR (101MHz, DMSO) δ168.90,168.12,164.28,161.83,151.76,150.95,135.40,132.78,132.75,131.52,130.06,12 9.98,123.64,121.17,117.40,116.05,115.84,110.49,107.46,105.01,62.20,55.37,55.29,53.44,49.98,46.25.

[0151] Example 6: N-{6-[(1H-indazol-5-yl)amino]-2-{4-[4-(dimethylamino)benzoyl]piperazin-1-yl}pyridin-3-yl}-2-(pyrrolidin-1-yl)acetamide (6);

[0152] The synthetic route is as follows:

[0153]

[0154] Step A9: Synthesis of Intermediate A-IX

[0155] According to the method of step A4 in Example 1, intermediate A-IV was used as a raw material to undergo acylation reaction with 4-dimethylaminobenzoyl chloride to obtain intermediate A-IX in a yield of 89.5%.

[0156] Step A10: Synthesis of Intermediate A-X

[0157] According to the method of step A6 in Example 1, intermediate A-IX was used as a raw material to undergo a nucleophilic substitution reaction with intermediate A-II to obtain intermediate A-X with a yield of 49.6%. Analytical data: ESI-MS [M+H] + (m / z):571.3.

[0158] Step A11: Synthesis of Intermediate A-XI

[0159] According to the method of step A7 in Example 1, intermediate A-X is used as a raw material, and a reduction reaction is carried out under the action of hydrogen and palladium on carbon to obtain intermediate A-XI with a yield of 82.3%.

[0160] Step A12: Synthesis of Intermediate A-XII

[0161] According to the method of step A8 in Example 1, intermediate A-XI was used as a raw material and subjected to acylation reaction with chloroacetyl chloride to obtain intermediate A-XII with a yield of 74.3%.

[0162] Step A13: Synthesis of Example 6

[0163] According to the method of step A9 in Example 1, intermediate A-XII was used as a raw material, and after a nucleophilic substitution reaction with tetrahydropyrrole, deprotection was performed under acidic conditions to obtain Example 6, with a yield of 37.5%. Analytical data: ESI-MS [M+H] + (m / z):568.1. 1H NMR (400MHz, DMSO-d6) δ12.84(s,1H),9.31(s,1H),8.84(s,1H),8.26(d,J=8.6Hz,1H),8 .11(t,J=1.4Hz,1H),7.93(s,1H),7.46-7.42(m,2H),7.35(s,1H),7.33(d,J=2.0Hz,1H), 6.74(d,J=2.1Hz,1H),6.73(d,J=2.1Hz,1H),6.55(d,J=8.7Hz,1H),3.70(t,J=4.7Hz,4H ),3.26(s,2H),3.08(t,J=5.0Hz,4H),2.96(s,6H),2.74-2.63(m,4H),1.82-1.78(m,4H). 13 C NMR (101MHz, DMSO) δ170.44,168.56,151.73,150.92,136.39,135.45,133.25,131.09,129.51,123.63 ,122.48,121.12,117.58,111.54,110.52,107.35,105.04,59.88,55.37,54.68,50.30,45.86,24.22.

[0164] Example 7: N-{6-[(1H-indazol-5-yl)amino]-2-{4-[4-(dimethylamino)benzoyl]piperazin-1-yl}pyridin-3-yl}-2-(dimethylamino)acetamide; (7);

[0165] According to the method of step A9 in Example 1, intermediate A-XII was used as a raw material, and dimethylamine hydrochloride was reacted with the intermediate A-XII to undergo nucleophilic substitution reaction, followed by deprotection under acidic conditions to obtain Example 7, with a yield of 29.9%. Analytical data: ESI-MS [M+Na] + (m / z): 564.4.1H NMR (400MHz, DMSO-d6) δ12.83(s,1H),9.33(s,1H),8.84(s,1H),8.24(d,J=8.6Hz,1H),8.11(s,1H),7.93(s,1H),7.44(s,2H),7.35( d,J=8.3Hz,2H),6.73(d,J=8.3Hz,2H),6.54(d,J=8.7Hz,1H),3.72(s,4H),3.11-3.04(m,4H),2.96(s,6H),2.50(s,2H),2.35(s,6H). 13C NMR (101MHz, DMSO) δ170.41,168.26,151.72,151.00,136.39,135.46,133.27,131.09,129.56,123 .63,122.51,121.11,117.39,111.54,110.51,107.34,104.90,65.38,63.70,55.37,50.23,46.24.

[0166] Example 8: N-{6-[(1H-indazol-5-yl)amino]-2-{4-[4-(dimethylamino)benzoyl]piperazin-1-yl}pyridin-3-yl}-2-(cyclopropylamino)acetamide (8);

[0167] According to the method of step A9 in Example 1, intermediate A-XII was used as a raw material, and after a nucleophilic substitution reaction with cyclopropylamine, deprotection was performed under acidic conditions to obtain Example 8 with a yield of 40.1%. Analytical data: ESI-MS [M+H] + (m / z):554.1. 1 H NMR (400MHz, DMSO-d6) δ12.83(s,1H),9.33(s,1H),8.82(s,1H),8.23(d,J=8.6Hz,1H),8.11(d,J=1 .5Hz,1H),7.93(s,1H),7.44(d,J=2.0Hz,2H),7.37-7.31(m,2H),6.77-6.70(m,2H),6.53(d,J=8.6 Hz,1H),4.03(q,J=7.1Hz,1H),3.73-3.68(m,4H),3.08(t,J=4.9Hz,4H),2.96(s,6H),2.21(tt,J=6 .7,3.6Hz,1H),1.99(s,1H),1.18(t,J=7.1Hz,1H),0.49-0.38(m,2H),0.35(dd,J=4.6,2.3Hz,2H). 13 C NMR (101MHz, DMSO) δ170.45,170.03,151.73,151.66,151.03,136.38,135.48,133.26,131.21,129.52,123.6 4,122.53,121.11,117.38,111.55,110.49,107.33,104.79,60.21,53.37,50.05,31.32,21.22,14.55,6.65.

[0168] Example 9: N-{6-[(1H-indazol-5-yl)amino]-2-{4-[4-(dimethylamino)benzoyl]piperazin-1-yl}pyridin-3-yl}-2-[(2-hydroxyethyl)amino]acetamide (9);

[0169] According to the method of step A9 in Example 1, intermediate A-XII was used as a raw material, and after nucleophilic substitution reaction with ethanolamine, deprotection was carried out under acidic conditions to obtain Example 9 with a yield of 23.5%. Analytical data: ESI-MS [M+H] + (m / z):558.0. 1 H NMR(400MHz,DMSO-d6)δ12.83(s,1H),9.59(s,1H),8.82(s,1H),8.24(d,J=8.6Hz,1H), 8.11(s,1H),7.93(s,1H),7.44(s,2H),7.34(d,J=8.4Hz,2H),6.73(d,J=8.4Hz,2H),6.5 3(d,J=8.7Hz,1H),4.62(d,J=5.3Hz,1H),3.73(t,J=4.7Hz,4H),3.54(q,J=5.7,5.2Hz,2 H), 3.29 (s, 2H), 3.18 (s, 1H), 3.10 (t, J = 5.0Hz, 4H), 2.96 (s, 6H), 2.69 (t, J = 5.7Hz, 2H). 13 C NMR (101MHz, DMSO) δ170.39,169.99,151.71,151.63,151.10,136.42,135.51,133.01,131.22,129.53,12 3.64,122.58,121.11,117.37,111.54,110.50,107.29,104.68,60.96,53.09,52.34,50.03,49.07,40.26.

[0170] Example 10: N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-methylbenzoyl)piperazin-1-yl]pyridin-3-yl}-2-[(2-hydroxyethyl)amino]acetamide (10);

[0171] The synthetic route is as follows:

[0172]

[0173] Step A14: Synthesis of Intermediate A-XIII

[0174] According to the method of step A4 in Example 1, intermediate A-IV was used as a raw material to undergo acylation reaction with 4-methylbenzoyl chloride to obtain intermediate A-XIII in a yield of 92.3%.

[0175] Step A15: Synthesis of Intermediate A-XIV

[0176] According to the method of step A6 in Example 1, intermediate A-XIII is used as a raw material to undergo nucleophilic substitution reaction with intermediate A-II to obtain intermediate A-XIV in a yield of 41.2%.

[0177] Step A16: Synthesis of Intermediate A-XV

[0178] According to the method of step A7 in Example 1, intermediate A-XIII is used as a raw material, and a reduction reaction is carried out under the action of hydrogen and palladium on carbon to obtain intermediate A-XV with a yield of 88.2%.

[0179] Step A17: Synthesis of Intermediate A-XVI

[0180] According to the method of step A8 in Example 1, intermediate A-XV was used as a raw material to undergo acylation reaction with chloroacetyl chloride to obtain intermediate A-XVI in a yield of 79.1%.

[0181] Step A18: Synthesis of Example 10

[0182] According to the method of step A9 in Example 1, intermediate A-XVI was used as a raw material, and after nucleophilic substitution reaction with ethanolamine, deprotection was carried out under acidic conditions to obtain Example 10 with a yield of 52.1%. Analytical data: ESI-MS [M+H] + (m / z):529.0. 1 H NMR(400MHz,DMSO-d6)δ12.84(s,1H),9.58(s,1H),8.83(s,1H),8.19(d,J =8.6Hz,1H),8.11(s,1H),7.93(s,1H),7.44(s,2H),7.36(d,J=7.8Hz,2H) ,7.28(d,J=7.7Hz,2H),6.53(d,J=8.6Hz,1H),4.65(s,1H),3.77(m,5H),3 .53(q,J=6.5,6.1Hz,2H),3.11(s,4H),2.70(t,J=5.6Hz,2H),2.36(s,3H). 13C NMR (101MHz, DMSO) δ169.79,169.72,151.73,151.20,139.68,136.46,135.48,133.53,133.19,131.60,12 9.38,127.56,123.64,121.15,117.13,110.53,107.38,104.62,60.76,55.36,52.87,52.20,49.90,21.38.

[0183] Example 11: N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-methylbenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(4-hydroxypiperidin-1-yl)acetamide (11);

[0184] According to the method of step A9 in Example 1, intermediate A-XVI was used as a raw material, and after a nucleophilic substitution reaction with 4-hydroxypiperidine, deprotection was performed under acidic conditions to obtain Example 11 with a yield of 55.7%. Analytical data: ESI-MS [M+H] + (m / z):569.1. 1 H NMR (400MHz, DMSO-d6) δ12.84(s,1H),9.28(s,1H),8.84(s,1H),8.26(d,J=8.7Hz,1H),8.11(s,1H),7 .92(s,1H),7.49-7.40(m,2H),7.36(d,J=7.8Hz,2H),7.28(d,J=7.8Hz,2H),6.55(d,J=8.7Hz,1H),4. 68(d,J=4.2Hz,1H),3.72(s,4H),3.56-3.50(m,1H),3.38(q,J=7.1Hz,1H),3.09(s,4H),2.79(dt,J=1 0.1, 4.3Hz, 2H), 2.36 (s, 3H), 2.30 (t, J = 10.9Hz, 2H), 1.79 (dq, J = 12.2, 3.8Hz, 2H), 1.57-1.45 (m, 2H). 13 C NMR (101MHz, DMSO) δ169.79,168.35,151.71,150.87,139.74,136.42,135.42,133.44,133.25,131.21,129.4 2,127.53,123.64,121.18,117.61,110.48,107.47,105.11,66.04,65.37,62.24,51.96,50.20,35.03,21.38.

[0185] Example 12: N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-methylbenzoyl)piperazin-1-yl]pyridin-3-yl}-2-[4-(2-hydroxyethyl)piperazin-1-yl]acetamide (12);

[0186] According to the method of step A9 in Example 1, intermediate A-XVI was used as a raw material, and after a nucleophilic substitution reaction with N-hydroxyethylpiperazine, deprotection was performed under acidic conditions to obtain Example 12 with a yield of 46.5%. Analytical data: ESI-MS [M+H] + (m / z):598.0. 1 H NMR (400MHz, DMSO-d6) δ12.84(s,1H),9.21(s,1H),8.84(s,1H),8.24(d,J=8.6Hz ,1H),8.11(s,1H),7.93(s,1H),7.44(d,J=2.9Hz,2H),7.36(d,J=7.7Hz,2H),7.2 8(d,J=7.7Hz,2H),6.55(d,J=8.7Hz,1H),4.43(s,1H),3.70(s,4H),3.52(d,J=6. 0Hz, 2H), 3.10 (d, J = 8.6Hz, 4H), 2.55 (s, 8H), 2.44 (t, J = 6.1Hz, 2H), 2.36 (s, 3H). 13 C NMR (101MHz, DMSO) δ169.89,168.09,151.75,150.92,139.75,136.46,135.42,133.47,133.25,131.36,129.4 4,127.49,123.63,121.17,117.52,110.50,107.46,105.05,62.27,60.71,58.90,53.72,53.51,50.11,21.38.

[0187] Example 13: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-(methylsulfonamido)pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate (13);

[0188] The synthetic route is as follows:

[0189]

[0190] Step B1: Synthesis of intermediate B-Ⅰ

[0191] Dissolve 2,6-dichloro-3-nitropyridine (15.97 g, 83.0 mmol) in tetrahydrofuran (150 mL), add potassium carbonate (28.7 g, 207.6 mmol) and L-prolinol (7 g, 69.2 mmol), and stir at room temperature for 12 h. Evaporate the reaction solution to dryness, add water (200 mL) and stir, filter and dry to obtain a crude product, which is purified by column chromatography (PE:EA=3:1) to obtain 15.2 g of a yellow solid with a yield of 85.2%. Analytical data: 1 H NMR (400MHz, DMSO-d6) δ8.23(d,J=8.4Hz,1H),6.82(d,J=8.3Hz,1H),4.79(s,1H),4.31(qd,J=7.0,3.3Hz,1H),3.65(dd,J=10.6,3.4Hz,1H), 3.45(dd,J=10.6,6.3Hz,1H),3.40-3.26(m,1H),2.78(ddd,J=10.6,7.8,2.5Hz,1H),2.12-2.02(m,1H),2.01-1.86(m,2H),1.80-1.63(m,1H).

[0192] Step B2: Synthesis of intermediate B-II

[0193] 4-Dimethylaminobenzoic acid (0.15 g, 0.91 mmol) was dissolved in dry toluene (3 mL), triethylamine (0.28 g, 2.8 mmol) and diphenylphosphoryl azide (0.5 g, 1.8 mmol) were added, and the mixture was stirred at 50 °C for 45 min, and then intermediate B-I (0.47 g, 1.8 mmol) was added, and the mixture was stirred at 100 °C for 4 h. The reaction solution was evaporated to dryness, stirred with water (10 mL), extracted with dichloromethane (10 mL), and the organic phase was washed with 1.2 M hydrochloric acid aqueous solution (5 mL), dried over anhydrous sodium sulfate, and evaporated to dryness to obtain a crude product, which was purified by column chromatography (PE:EA=4:1) to obtain 0.23 g of a yellow solid with a yield of 70%. ESI-MS [M+Na] + (m / z):442.0.

[0194] Step B3: Synthesis of intermediate B-III

[0195] According to the method of step A6 in Example 1, intermediate B-Ⅱ was used as a raw material to undergo nucleophilic substitution reaction with intermediate A-Ⅱ to obtain intermediate B-Ⅲ with a yield of 51.2%.

[0196] Step B4: Synthesis of intermediate B-IV

[0197] According to the method of step A7 in Example 1, intermediate B-III was used as a raw material, and a reduction reaction was carried out under the action of hydrogen and palladium on carbon to obtain intermediate B-IV with a yield of 73.1%.

[0198] Step B5: Synthesis of Example 13

[0199] According to the method of step A8 in Example 1, intermediate B-IV was used as a raw material, and acylated with methanesulfonyl chloride, and then deprotected under acidic conditions to obtain Example 13 with a yield of 43.9%. Analytical data: ESI-MS [M+H] + (m / z):565.0. 1 H NMR(400MHz,DMSO-d6)δ12.79(s,1H),9.26(s,1H),8.79(s,1H),8.56(s,1H),8.11(s,1H),7.93(s,1H),7.40(s,2H),7.23(s,3H),6. 65(s,2H),6.14(s,1H),4.51(d,J=173.8Hz,2H),3.91(s,2H),3.47(s,1H),2.80(s,6H),2.01(s,2H),1.93-1.73(m,3H),1.23(s,2H).

[0200] Example 14: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl{4-(dimethylamino)phenyl}carbamate (14);

[0201]

[0202] According to the method of step A9 in Example 1, intermediate B-V was used as a raw material, and after a nucleophilic substitution reaction with tetrahydropyrrole, deprotection was performed under acidic conditions to obtain Example 14 with a yield of 39.5%. Analytical data: ESI-MS [M+H] + (m / z):598.0. 1H NMR(400MHz,DMSO-d6)δ12.79(s,1H),9.17(d,J=81.6Hz,2H),8.71(s,1H),8.12(s,1 H),7.92(s,1H),7.43(s,1H),7.40(s,2H),7.28-7.22(m,2H),6.65(d,J=8.5Hz,2H), 6.24(d,J=8.4Hz,1H),4.68-4.23(m,2H),3.91(s,1H),3.65(d,J=8.4Hz,1H),3.21(s ,3H),2.81(s,6H),2.07(s,0H),1.97(s,1H),1.85(s,2H),1.73(s,4H),1.23(s,2H). 13 C NMR(151MHz,DMSO-d6)δ162.78,154.29,153.28,152.69,147.21,135.47,130.12,123.62,121.06, 120.61,113.37,110.50,107.10,100.04,57.17,54.52,50.48,41.08,29.50,28.67,24.55,23.53.

[0203] Example 15: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(dimethylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate (15);

[0204] According to the method of step A9 in Example 1, intermediate B-V was used as a raw material, and dimethylamine hydrochloride was subjected to nucleophilic substitution reaction, and then deprotected under acidic conditions to obtain Example 15 with a yield of 23.4%. Analytical data: ESI-MS [M+H] + (m / z):572.0. 1H NMR(400MHz,DMSO-d6)δ12.79(s,1H),9.29(s,1H),9.02(s,1H),8.72(s,1H),8.14-8.07(m,1H), 7.91(s,1H),7.42(s,1H),7.36(d,J=8.4Hz,2H),7.24(s,2H),6.69-6.62(m,2H),6.21(d,J=8.4H z,1H),4.57(d,J=12.2Hz,1H),4.29(dd,J=10.2,3.8Hz,1H),3.90(t,J=9.2Hz,1H),3.68(dd,J=9 .8,6.6Hz,1H),3.25(dt,J=9.3,6.6Hz,1H),3.03(s,2H),2.81(s,6H),2.28(s,6H),1.23(s,4H).

[0205] Example 16: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-methylpiperazin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate (16);

[0206] According to the method of step A9 in Example 1, intermediate B-V was used as a raw material, and after a nucleophilic substitution reaction with N-methylpiperazine, deprotection was performed under acidic conditions to obtain Example 37 with a yield of 40.5%. Analytical data: ESI-MS [M+H] + (m / z):627.0. 1 H NMR(400MHz,DMSO-d6)δ12.81(s,1H),9.29(s,1H),9.12(s,1H),8.76(s,1H),8.13(s,1H),7.9 1(s,1H),7.45(d,J=8.1Hz,1H),7.40(s,2H),7.24(d,J=8.5Hz,2H),6.65(d,J=8.5Hz,2H),6.25 (d,J=8.3Hz,1H),4.58(t,J=6.3Hz,1H),4.28(dd,J=10.4,4.1Hz,1H),3.89(t,J=9.3Hz,1H),3 .74-3.52(m,2H),3.23-3.07(m,2H),2.81(s,6H),2.50(s,3H),2.38(d,8H),2.11-1.81(m,4H).

[0207] Example 17: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[(cyclopropylmethyl)amino]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate (17);

[0208] According to the method of step A9 in Example 1, intermediate B-V was used as a raw material, and after a nucleophilic substitution reaction with cyclopropylmethylamine, deprotection was performed under acidic conditions to obtain Example 17 with a yield of 32.4%. Analytical data: ESI-MS [M+H] + (m / z):598.2. 1 H NMR(400MHz,DMSO-d6)δ12.67(s,1H),9.16(s,1H),9.09(s,1H),8.60(s,1H),8.00(s,1H) ,7.79(s,1H),7.31(s,1H),7.27(s,2H),7.12(s,2H),6.53(d,J=8.3Hz,2H),6.11(d,J=7.8 Hz,1H),4.48(s,1H),4.18(s,1H),3.77(s,1H),3.59(s,1H),3.19(s,2H),3.12-2.92(m,2H ),2.69(s,6H),1.91(d,J=38.9Hz,2H),1.12(s,4H),0.77(s,1H),0.15(d,J=117.1Hz,4H).

[0209] Example 18: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[(2-hydroxyethyl)amino]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate (18);

[0210] According to the method of step A9 in Example 1, intermediate B-V was used as a raw material, and after nucleophilic substitution reaction with ethanolamine, deprotection was carried out under acidic conditions to obtain Example 42 with a yield of 40.5%. Analytical data: ESI-MS [M+H] + (m / z):588.2.

[0211] 1H NMR(400MHz,DMSO-d6)δ12.82(s,1H),9.43(s,1H),9.33(s,1H),8.78(s,1H),8.13( s,1H),7.92(s,1H),7.39(s,1H),7.26(s,2H),7.11(s,2H),6.65(s,2H),6.21(s,1H) ,5.32(s,1H),4.87(s,1H),4.63-4.59(m,1H),4.32(s,1H),3.87-3.83(m,1H),3.70( s,2H),3.54(s,2H),2.80(s,6H),1.98(s,2H),1.85-1.81(m,2H),0.87-0.82(m,4H).

[0212] Example 19: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-acrylamidopyridin-2-yl}pyrrolidin-2-yl}p-tolylcarbamate (19);

[0213] The synthetic route is as follows:

[0214]

[0215] Step B6: Synthesis of Intermediate B-VI

[0216] According to the method of step B2 in Example 13, p-toluic acid and intermediate B-I were used as raw materials to undergo Curtis rearrangement reaction to produce intermediate B-VI with a yield of 65.8%. Analytical data: ESI-MS [M+H] + (m / z):391.9.

[0217] Step B7: Synthesis of Intermediate B-VII

[0218] According to the method of step A6 in Example 1, intermediate B-VI was used as a raw material to undergo a nucleophilic substitution reaction with intermediate A-II to obtain intermediate B-VII with a yield of 41.9%. :ESI-MS [M+H] + (m / z):572.0.

[0219] Step B8: Synthesis of Intermediate B-VIII

[0220] According to the method of step A7 in Example 1, intermediate B-VII is used as a raw material, and a reduction reaction is carried out under the action of hydrogen and palladium on carbon to obtain intermediate B-VIII with a yield of 89.4%.

[0221] Step B9: Synthesis of Example 19

[0222] According to the method of step A8 in Example 1, intermediate B-VIII was used as a raw material, and acryloyl chloride was subjected to acylation reaction, followed by deprotection under acidic conditions to obtain Example 19 with a yield of 62.7%. Analytical data: ESI-MS [M+H]+ (m / z): 512.0.

[0223] Example 20: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-methylpiperazin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}p-tolylcarbamate (20);

[0224]

[0225] According to the method of step A9 in Example 1, intermediate B-IX was used as a raw material, and after a nucleophilic substitution reaction with 4-methylpiperazine, deprotection was performed under acidic conditions to obtain Example 20 with a yield of 42.5%. Analytical data: ESI-MS [M+H] + (m / z):598.1. 1 H NMR(400MHz,DMSO-d6)δ12.79(s,1H),9.54(s,1H),9.09(s,1H),8.73(s,1H),8.11(s,1H),7.91(s,1H), 7.60(d,J=8.4Hz,1H),7.39(d,J=2.9Hz,2H),7.31(d,J=8.0Hz,2H),7.05(d,J=8.1Hz,2H),6.28(d,J=8.4 Hz,1H),4.58(t,J=5.9Hz,1H),4.28(dd,J=10.4,4.2Hz,1H),3.99-3.90(m,1H),3.75-3.45(m,2H),3.22 -3.15(m,2H),3.05(s,2H),2.36(s,4H),2.22(s,3H),2.16(s,3H),2.13-2.05(m,2H),2.02-1.78(m,4H). 13 C NMR(151MHz,DMSO-d6)δ169.02,154.04,152.47,151.65,137.01,136.20,135.56,133.27,131.63,129.52,123.63,12 1.05,118.83,112.49,110.39,107.00,101.16,65.84,61.87,57.33,55.19,53.37,51.18,46.17,28.67,24.55,20.79.

[0226] Example 21: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopentylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}p-tolylcarbamate (21);

[0227] According to the method of step A9 in Example 1, intermediate B-IX was used as a raw material, and after nucleophilic substitution reaction with cyclopentylamine, deprotection was performed under acidic conditions to obtain Example 21 with a yield of 38.2%. Analytical data: ESI-MS [M+H] + (m / z):583.3. 1 H NMR(400MHz,DMSO-d6)δ12.77(s,1H),9.55(s,1H),9.22(s,1H),8.71(s,1H),8.10(s,1H),7.90(s,1H),7.52 (d,J=8.3Hz,1H),7.38(d,J=2.6Hz,2H),7.32(d,J=8.0Hz,2H),7.05(d,J=8.1Hz,2H),6.25(d,J=8.4Hz,1H), 4.59(s,1H),4.30(dd,J=10.3,4.0Hz,1H),3.91(t,J=9.2Hz,1H),3.70(d,J=8.3Hz,1H),3.23(d,J=6.3Hz,2H ),3.19-3.04(m,2H),3.02(s,1H),2.22(s,3H),2.16-1.88(m,4H),1.86-1.65(m,4H),1.54(d,J=60.8Hz,4H).

[0228] Example 22: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-hydroxypiperidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl p-tolylcarbamate (22);

[0229] According to the method of step A9 in Example 1, intermediate B-IX was used as a raw material, and after a nucleophilic substitution reaction with 4-hydroxypiperidine, deprotection was performed under acidic conditions to obtain Example 22 with a yield of 41.9%. Analytical data: ESI-MS [M+H] + (m / z):599.3. 1H NMR(400MHz,DMSO-d6)δ12.79(s,1H),9.53(s,1H),9.11(s,1H),8.73(s,1H),8.11(s,1H),7.90 (s,1H),7.58(d,J=8.3Hz,1H),7.39(s,2H),7.34-7.23(m,2H),7.05(d,J=7.9Hz,2H),6.35-6.18 (m,1H),4.60(s,1H),4.29(s,1H),3.80(d,J=92.2Hz,1H),3.48(s,1H),3.18(s,1H),3.02(s,2H ),2.73(s,2H),2.22(s,3H),1.93(d,J=49.6Hz,4H),1.59(d,J=106.2Hz,4H),1.26-0.83(m,4H).

[0230] Example 23: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-ethylpiperazin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl p-tolylcarbamate (23);

[0231] According to the method of step A9 in Example 1, intermediate B-IX was used as a raw material, and after a nucleophilic substitution reaction with N-ethylpiperazine, deprotection was performed under acidic conditions to obtain Example 23 with a yield of 40.5%. Analytical data: ESI-MS [M+H] + (m / z): 612.3. 1 H NMR (400MHz, DMSO-d6) δ12.79(s,1H),9.54(s,1H),9.10(s,1H),8.74(s,1H),8.11(s,1H),7.91(s,1H),7.61(d,J=8.4Hz,1 H),7.42-7.37(m,2H),7.31(d,J=8.0Hz,2H),7.05(d,J=8.2Hz,2H),6.28(d,J=8.4Hz,1H),4.60-4.56(m,1H),4.28(dd,J=10 .3,4.2Hz,1H),3.95(dd,J=10.4,7.7Hz,1H),3.69(dt,J=9.9,6.4Hz,1H),3.22-3.12(m,2H),3.07-3.03(m,2H),2.52(s,2H) ,2.42(s,2H),2.32(d,J=7.1Hz,2H),2.21(s,3H),2.14-1.97(m,2H),1.93-1.80(m,2H),1.23(s,1H),0.97(t,J=7.1Hz,3H).13 C NMR(151MHz,DMSO-d6)δ169.01,154.03,152.46,151.64,137.02,136.25,135.57,133.27,131.62,129.51,123.64,12 1.05,118.81,110.39,107.00,101.17,65.80,61.90,57.34,53.46,52.81,52.05,51.19,28.66,24.55,20.79,12.32.

[0232] Example 24: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[4-(2-hydroxyethyl)piperazin-1-yl]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}p-tolylcarbamate (24);

[0233] According to the method of step A9 in Example 1, intermediate B-IX was used as a raw material, and after nucleophilic substitution reaction with 4-hydroxyethylpiperazine, deprotection was performed under acidic conditions to obtain Example 50 with a yield of 40.5%. Analytical data: ESI-MS [M+H] + (m / z): 628.2. 1 H NMR(400MHz,DMSO-d6)δ12.81(s,1H),9.72-9.52(m,1H),9.44(s,1H),9.13(s,1H),8.77(s,1H), 8.12(s,1H),7.90(s,1H),7.50(s,2H),7.39(s,2H),7.34-7.20(m,2H),7.04(d,J=8.1Hz,1H),6. 26(s,1H),4.58(s,1H),4.37-4.22(m,1H),3.93(s,1H),3.68(s,1H),3.60(s,2H),3.58(s,2H),3 .10(s,2H),2.21(s,3H),1.85(t,,J=9Hz,2H),1.65(d,J=42.4Hz,4H),1.75(s,4H),1.15(d,4H).

[0234] Example 25: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl (4-fluorophenyl)carbamate (25);

[0235] The synthetic route is as follows:

[0236]

[0237] Step B10: Synthesis of Intermediate B-X

[0238] According to the method of step B2 in Example 13, p-fluorobenzoic acid and intermediate B-I were used as raw materials to undergo Curtis rearrangement reaction to produce intermediate B-X with a yield of 62.9%. Analytical data: ESI-MS [MH] - (m / z):392.9.

[0239] Step B11: Synthesis of Intermediate B-XI

[0240] According to the method of step A6 in Example 1, intermediate B-X was used as a raw material to undergo a nucleophilic substitution reaction with intermediate A-II to obtain intermediate B-XI with a yield of 43.5%. Analytical data: ESI-MS [M+H] + (m / z):575.9.

[0241] Step B12: Synthesis of Intermediate B-XII

[0242] According to the method of step A7 in Example 1, intermediate B-XI is used as a raw material, and a reduction reaction is carried out under the action of hydrogen and palladium on carbon to obtain intermediate B-XII with a yield of 92.4%.

[0243] Step B13: Synthesis of Intermediate B-XIII

[0244] According to the method of step A8 in Example 1, intermediate B-XII was used as a raw material and subjected to acylation reaction with chloroacetyl chloride to obtain intermediate B-XIII with a yield of 89.3%.

[0245] Step B14: Synthesis of Example 25

[0246] According to the method of step A9 in Example 1, intermediate B-XIII was used as a raw material, and after a nucleophilic substitution reaction with tetrahydropyrrole, deprotection was performed under acidic conditions to obtain Example 25 with a yield of 51.2%. Analytical data: ESI-MS [M+H] + (m / z):573.0. 1H NMR (400MHz, DMSO-d6) δ12.78(s,1H),9.73-9.68(m,1H),9.05(s,1H),8.72(s,1H),8.11(d,J=1.5Hz,1H),7.9 3(d,J=22.3Hz,1H),7.46(s,1H),7.44(s,2H),7.39(d,J=1.9Hz,2H),7.14-7.05(m,2H),6.24(d,J=8.4Hz,1H) ,4.58(tt,J=7.7,4.5Hz,1H),4.37-4.26(m,1H),3.96(dd,J=10.3,7.7Hz,1H),3.65(dt,J=9.6,6.3Hz,1H),3. 24(dd,J=9.6,6.8Hz,1H),3.18(s,2H),2.58(p,J=3.8Hz,4H),2.17-1.79(m,4H),1.72(td,J=4.6,2.1Hz,4H).

[0247] Example 26: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-pivalamidopyridin-2-yl}pyrrolidin-2-yl}methyl(4-fluorophenyl)carbamate (26);

[0248] According to the method of step A8 in Example 1, intermediate B-XII was used as a raw material, and acylated with pivaloyl chloride, followed by deprotection under acidic conditions to obtain Example 26 with a yield of 74.2%. Analytical data: ESI-MS [M+H] + (m / z):546.1. 1 H NMR(400MHz,DMSO-d6)δ12.77(s,1H),9.73(s,1H),8.69(s,2H),8.14(s,1H),7 .91(s,1H),7.45(s,2H),7.40(s,2H),7.11(d,J=8.9Hz,2H),7.03(d,J=8.3Hz, 1H),6.16(d,J=8.3Hz,1H),4.64(s,1H),4.39(d,J=10.1Hz,1H),3.91(s,1H),3 .70(d,J=9.1Hz,1H),3.34(s,2H),2.01(s,2H),1.92-1.80(m,2H),1.19(s,9H).

[0249] Example 27: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-hydroxypiperidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl (4-fluorophenyl)carbamate (27);

[0250] According to the method of step A9 in Example 1, intermediate B-XIII was used as a raw material, and after a nucleophilic substitution reaction with 4-hydroxypiperidine, deprotection was performed under acidic conditions to obtain Example 27 with a yield of 54.7%. Analytical data: ESI-MS [M+H] + (m / z): 603.2. 1 H NMR(400MHz,DMSO-d6)δ12.79(s,1H),9.80-9.61(m,1H),9.11(s,1H),8.74(s,1H),8.11(s,1H),7.89(s,1H),7. 58(d,J=8.4Hz,1H),7.43(s,2H),7.39(s,2H),7.09(t,J=8.9Hz,2H),6.27(d,J=8.4Hz,1H),4.61(d,J=4.1Hz,1H) ,4.58(d,J=9.0Hz,1H),4.30(dd,J=10.3,4.1Hz,1H),3.95(dd,J=10.4,7.7Hz,1H),3.69(dt,J=9.7,6.3Hz,1H), 3.53-3.45(m,1H),3.24-3.14(m,1H),3.02(s,2H),2.32-2.02(m,4H),1.99-1.76(m,4H),1.60(d,J=96.4Hz,4H).

[0251] Example 28: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[(cyclopropylmethyl)amino]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}methyl (4-fluorophenyl)carbamate (28);

[0252] According to the method of step A9 in Example 1, intermediate B-XIII was used as a raw material, and after nucleophilic substitution reaction with cyclopropylmethylamine, deprotection was performed under acidic conditions to obtain Example 28 with a yield of 49.6%. Analytical data: ESI-MS [M+H] + (m / z):573.7. 1H NMR(400MHz,DMSO-d6)δ12.67(s,1H),9.59(s,1H),9.10(s,1H),8.60(s,1H) ,7.98(s,1H),7.77(s,1H),7.32(d,3H),7.28(s,2H),6.97(s,2H),6.22-6.0 3(m,1H),4.49(s,2H),3.81(s,1H),3.58(s,1H),3.21(s,2H),3.09(s,2H),1 .96(d,2H),1.85(s,2H),1.73(s,2H),1.46-1.30(d,1H),0.29-0.10(s,4H). 13 C NMR(151MHz,DMSO-d6)δ170.52,157.31,156.48,154.12,152.74,152.25,137.52,135.90,135.56,133.18,130.12,123.61 ,121.06,120.46,115.76,115.61,110.97,106.99,100.44,65.93,57.15,54.24,52.12,50.84,29.48,28.65,11.05,3.69.

[0253] Example 29: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylpiperidin-4-ylcarbamate (29);

[0254] The synthetic route is as follows:

[0255]

[0256] Step B15: Synthesis of Intermediate B-XIV

[0257] According to the method of step B2 in Example 13, 1-Boc-4-piperidinic acid and intermediate B-I were used as raw materials to undergo Curtis rearrangement reaction to produce intermediate B-XIV with a yield of 52.6%. Analytical data: ESI-MS [M+Na] + (m / z):506.2. 1HNMR(400MHz,DMSO-d6)δ12.83(s,1H),9.05(s,1H),8.72(s,1H),8.12(s,1H),7.91(s,1H),7.4 1(d,J=8.2Hz,3H),7.19(d,J=7.8Hz,1H),6.23(d,J=8.4Hz,1H),4.71-4.12(m,2H),3.73(t,J=9. 4Hz,1H),3.62(t,J=6.4Hz,0H),3.56(t,7H),3.20(s,2H),2.93(d,J=12.2Hz,2H),2.63-2.58(m, 4H), 2.11-2.01 (m, 1H), 1.88 (ddd, J=52.0, 12.2, 6.1Hz, 2H), 1.75 (t, J=5.7Hz, 4H), 1.67 (d, 2H).

[0258] Step B16: Synthesis of Intermediate B-XV

[0259] According to the method of step A6 in Example 1, intermediate B-XIV was used as a raw material to undergo a nucleophilic substitution reaction with intermediate A-II to obtain intermediate B-XV with a yield of 49.8%. Analytical data: ESI-MS [M+H] + (m / z): 665.2.

[0260] Step B17: Synthesis of Intermediate B-XVI

[0261] According to the method of step A7 in Example 1, intermediate B-XV is used as a raw material, and a reduction reaction is carried out under the action of hydrogen and palladium on carbon to obtain intermediate B-XVI with a yield of 92.8%.

[0262] Step B18: Synthesis of Intermediate B-XVII

[0263] According to the method of step A8 in Example 1, intermediate B-XVI was used as a raw material and acylated with chloroacetyl chloride to obtain intermediate B-XVII with a yield of 85.1%. Analytical data: ESI-MS [M+Na] + (m / z): 733.4

[0264] Step B19: Synthesis of Example 29

[0265] According to the method of step A9 in Example 1, intermediate B-XVII was used as a raw material, and after a nucleophilic substitution reaction with tetrahydropyrrole, deprotection was performed under acidic conditions to obtain Example 29 with a yield of 36.7%. Analytical data: ESI-MS [M+H] + (m / z):562.5. 1H NMR(400MHz,DMSO-d6)δ12.83(s,1H),9.05(s,1H),8.72(s,1H),8.12(s,1H),7.91(s,1H),7.41 (d,J=8.2Hz,3H),7.19(d,J=7.8Hz,1H),6.23(d,J=8.4Hz,1H),4.71-4.12(m,2H),3.73(t,J=9.4 Hz,1H),3.62(t,J=6.4Hz,1H),3.56(t,4H),3.20(s,2H),2.93(d,J=12.2Hz,2H),2.63-2.58(m, 4H),2.11-2.01(m,1H),1.88(ddd,J=52.0,12.2,6.1Hz,4H),1.75(t,J=5.7Hz,6H),1.67(d,2H).

[0266] Example 30: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-(2-thiomorpholinoacetylamino)pyridin-2-yl}pyrrolidin-2-yl}methylpiperidin-4-ylcarbamate (30);

[0267] According to the method of step A9 in Example 1, intermediate B-XVII was used as a raw material, and after nucleophilic substitution reaction with thiomorpholine, deprotection was carried out under acidic conditions to obtain Example 61 with a yield of 45.6%. Analytical data: ESI-MS [M+H] + (m / z):594.5.

[0268] Example 31: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-methylpiperidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylpiperidin-4-ylcarbamate (31);

[0269] According to the method of step A9 in Example 1, intermediate B-XVII was used as a raw material, and after a nucleophilic substitution reaction with 4-methylpiperidine, deprotection was performed under acidic conditions to obtain Example 31 with a yield of 40.2%. Analytical data: ESI-MS [M+H] + (m / z):590.5.

[0270] Example 32: (S)-1-{2-{{6-[(1H-indazol-5-yl)amino]-2-{2-[(piperidin-4-ylcarbamoyl)oxy]methyl}pyrrolidin-1-yl}pyridin-3-yl}amino-2-oxoethyl}piperidine-4-carboxylic acid methyl ester (32);

[0271] According to the method of step A9 in Example 1, intermediate B-XVII was used as a raw material, and after nucleophilic substitution reaction with methyl 4-piperidinic acid ester, deprotection was carried out under acidic conditions to obtain Example 32 with a yield of 37.4%. Analytical data: ESI-MS [M+H] + (m / z): 634.7.

[0272] Example 33: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl isopropylcarbamate (33);

[0273] The synthetic route is as follows:

[0274]

[0275] Step B20: Synthesis of Intermediate B-XVIII

[0276] According to the method of step B2 in Example 13, isobutyric acid and intermediate B-I were used as raw materials to undergo Curtis rearrangement reaction to produce intermediate B-XVIII with a yield of 63.7%. Analytical data: 1 H NMR (400MHz, DMSO-d6) δ8.24(d,J=8.4Hz,1H),6.99(d,J=7.7Hz,1H),6.86(d,J=8.3Hz,1H),4.57 (d,J=35.4Hz,1H),4.27(dd,J=11.0,5.3Hz,1H),4.04(dd,J=11.1,3.5Hz,1H),3.61-3.48(m,1H), 3.38-3.33(m,1H),2.75(ddd,J=10.4,7.4,2.7Hz,1H),2.08(t,J=7.7Hz,1H),1.98(ddp,J=9.9,6. 6,2.8Hz,1H),1.93-1.80(m,1H),1.74(dtt,J=11.4,9.5,7.2Hz,1H),1.03(dd,J=6.6,3.1Hz,6H).

[0277] Step B21: Synthesis of Intermediate B-XIX

[0278] According to the method of step A6 in Example 1, intermediate B-XVIII was used as a raw material to undergo a nucleophilic substitution reaction with intermediate A-II to obtain intermediate B-XIX with a yield of 55.4%. Analytical data: ESI-MS [M+H] + (m / z):524.2.

[0279] Step B22: Synthesis of Intermediate B-XX

[0280] According to the method of step A7 in Example 1, intermediate B-XX is used as a raw material, and a reduction reaction is carried out under the action of hydrogen and palladium on carbon to obtain intermediate B-XX with a yield of 93.4%.

[0281] Step B23: Synthesis of Intermediate B-XXI

[0282] According to the method of step A8 in Example 1, intermediate B-XX was used as a raw material and subjected to acylation reaction with chloroacetyl chloride to obtain intermediate B-XXI in a yield of 88.1%.

[0283] Step B24: Synthesis of Example 68

[0284] According to the method of step A9 in Example 1, intermediate B-XXI was used as a raw material, and after nucleophilic substitution reaction with tetrahydropyrrole, deprotection was performed under acidic conditions to obtain Example 33 with a yield of 49.1%. Analytical data: ESI-MS [M+H] + (m / z):521.8. 1 H NMR (400MHz, DMSO-d6) δ12.79(s,1H),9.05(s,1H),8.71(s,1H),8.13(s,1H),7.92(s,1H),7.40(t,J=4 .6Hz,3H),7.06(d,J=7.8Hz,1H),6.22(d,J=8.4Hz,1H),4.51(dt,J=8.4,4.5Hz,1H),4.20(dd,J=10.4, 4.1Hz,1H),3.72(t,J=9.3Hz,1H),3.63(dt,J=12.5,6.4Hz,1H),3.54(p,J=6.7Hz,1H),3.24(d,J=6.6H z,1H),3.18(s,2H),2.68-2.55(m,4H),2.06-1.79(m,4H),1.74(d,J=6.5Hz,4H),1.04(d,J=6.5Hz,6H). 13 C NMR(101MHz,DMSO)δ169.81,155.86,152.73,152.27,137.60,135.61,133.42,123.66,120.93,1 11.18,110.37,106.83,100.45,65.43,59.46,57.35,54.44,50.65,42.74,28.66,24.48,24.02.

[0285] Example 34: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopropylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl isopropylcarbamate (34);

[0286] According to the method of step A9 in Example 1, intermediate B-XXI was used as a raw material, and after nucleophilic substitution reaction with cyclopropylamine, deprotection was carried out under acidic conditions to obtain Example 34 with a yield of 41.2%. Analytical data: ESI-MS [M+H] + (m / z):507.8. 1 H NMR(400MHz,DMSO-d6)δ12.79(s,1H),9.05(s,1H),8.71(s,1H),8.13(s,1H),7.91(s,1H),7.43-7.31(m,3H) ,7.07(d,J=7.8Hz,1H),6.20(d,J=8.3Hz,1H),4.52(s,1H),4.21(dd,J=10.6,3.9Hz,1H),3.74-3.61(m,2H),3 .55(dq,J=13.8,6.9Hz,1H),3.38(q,J=7.0Hz,1H),3.29(s,2H),3.19(dt,J=9.5,6.4Hz,1H),2.18(tt,J=6.7 ,3.6Hz,1H),1.29(d,J=47.1Hz,4H),1.04(d,J=6.6Hz,6H),0.39(m,J=6.3,3.0Hz,2H),0.30(m,J=3.8Hz,2H).

[0287] Example 35: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopentylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl isopropylcarbamate (35);

[0288] According to the method of step A9 in Example 1, intermediate B-XXI was used as a raw material, and after nucleophilic substitution reaction with cyclopentylamine, deprotection was carried out under acidic conditions to obtain Example 35 with a yield of 38.4%. Analytical data: ESI-MS [M+H] + (m / z):535.8. 1H NMR (400MHz, DMSO-d6) δ12.78(s,1H),9.20(s,1H),8.71(s,1H),8.12(s,1H),7.91(s,1H),7.50(d,J=8.4Hz,1H),7.39(d,J=4.2Hz,2H) ,7.06(d,J=7.9Hz,1H),6.24(d,J=8.4Hz,1H),4.52(s,1H),4.20(dd,J=10.3,4.0Hz,1H),3.75-3.63(m,2H),3.54(dt,J=13.6,6.8Hz,1 H),3.22(d,J=5.2Hz,2H),3.15(dt,J=13.1,4.8Hz,1H),3.04(p,J=6.0Hz,1H),1.98(dtd,J=34.2,11.4,9.4,5.0Hz,2H),1.80(td,J=12 .1,10.3,6.2Hz,2H),1.76-1.61(m,4H),1.48(tq,J=10.4,6.0Hz,2H),1.35(dq,J=11.6,6.0Hz,2H),1.23(s,1H),1.04(d,J=6.6Hz,6H).

[0289] Example 36: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylcyclopropylcarbamate (36);

[0290] The synthetic route is as follows:

[0291]

[0292] Step B25: Synthesis of Intermediate B-XXII

[0293] According to the method of step B2 in Example 13, cyclopropylcarboxylic acid and intermediate B-I were used as raw materials to undergo Curtis rearrangement reaction to produce intermediate B-XXII in a yield of 66.2%. 1H NMR (400MHz, DMSO-d6) δ8.25(d,J=8.4Hz,1H),7.29(s,1H),6.86(d,J=8.4Hz,1H),4 .54(s,1H),4.32-4.27(t,1H),4.04(d,J=10.6Hz,1H),3.34(s,1H),2.74(ddd,J=10 .3,7.4,2.5Hz,1H),2.42(tq,J=7.0,3.6Hz,1H),2.08(s,1H),2.01-1.95(m,1H),1. 85(s,1H),1.82-1.66(m,1H),0.54(dqd,J=8.2,4.9,2.0Hz,2H),0.40-0.34(m,2H).

[0294] Step B26: Synthesis of Intermediate B-XXIII

[0295] According to the method of step A6 in Example 1, intermediate B-XXII was used as a raw material to undergo a nucleophilic substitution reaction with intermediate A-II to obtain intermediate B-XXIII with a yield of 48.9%. Analytical data: ESI-MS [M+Na] + (m / z):544.2.

[0296] Step B27: Synthesis of Intermediate B-XXIV

[0297] According to the method of step A7 in Example 1, intermediate B-XXIII is used as a raw material, and a reduction reaction is carried out under the action of hydrogen and palladium on carbon to obtain intermediate B-XXIV in a yield of 91.8%.

[0298] Step B28: Synthesis of Intermediate B-XXV

[0299] According to the method of step A8 in Example 1, intermediate B-XXIV was used as a raw material to undergo acylation reaction with chloroacetyl chloride to obtain intermediate B-XXV in a yield of 89.4%.

[0300] Step B29: Synthesis of Example 36

[0301] According to the method of step A9 in Example 1, intermediate B-XXV was used as a raw material, and after nucleophilic substitution reaction with tetrahydropyrrole, deprotection was performed under acidic conditions to obtain Example 36 with a yield of 50.3%. Analytical data: ESI-MS [M+H] + (m / z):519.5. 1H NMR (400MHz, DMSO-d6) δ12.80(s,1H),9.05(s,1H),8.71(s,1H),8.12(s,1H),7.89(s,1H),7.39(d,J=3. 9Hz,3H),7.37(s,1H),6.22(d,J=8.4Hz,1H),4.50(s,1H),4.20(d,J=11.5Hz,1H),3.74(t,J=9.3Hz,1H) ,3.63(dt,J=9.5,6.4Hz,1H),3.24(d,J=6.5Hz,1H),3.21(s,2H),2.63-2.59(m,4H),2.41(tq,J=7.0,3. 5Hz,1H),2.09-1.88(m,2H),1.85-1.77(m,2H),1.77-1.68(m,4H),0.56-0.46(m,2H),0.43-0.32(m,2H). 13 C NMR (151MHz, DMSO-d6) δ169.76,157.33,152.74,152.27,137.67,136.22,135.61,133.37,123.65,120. 94,110.36,106.81,100.43,65.54,59.44,57.30,54.45,50.64,28.64,24.51,24.00,23.38,6.38,6.19.

[0302] Example 37: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopropylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylcyclopropylcarbamate (37);

[0303] According to the method of step A9 in Example 1, intermediate B-XXV was used as a raw material, and after nucleophilic substitution reaction with cyclopropylamine, deprotection was carried out under acidic conditions to obtain Example 37 with a yield of 54.1%. Analytical data: ESI-MS [M+H] + (m / z):505.5. 1H NMR (400MHz, DMSO-d6) δ12.79(s,1H),9.04(s,1H),8.71(s,1H),8.12(s,1H),7.89(s,1H),7.38(d,J=3.7Hz,3H),7 .34(d,J=11.8Hz,1H),6.20(d,J=8.4Hz,1H),4.51(s,1H),4.22(d,J=10.2Hz,1H),3.72(s,1H),3.64(d,J=11.5Hz,1 H),3.29(s,2H),3.22-2.81(m,2H),2.42(tq,J=7.0,3.6Hz,1H),2.17(tt,J=6.8,3.6Hz,1H),2.08-1.90(m,2H),1.8 0(dd,J=12.8,6.4Hz,2H),0.56-0.46(m,2H),0.45-0.39(m,2H),0.36(dd,J=5.6,3.0Hz,2H),0.30(q,J=3.3Hz,2H).

[0304] Example 38: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylcyclohexylcarbamate (38);

[0305] The synthetic route is as follows:

[0306]

[0307] Step B30: Synthesis of Intermediate B-XXVI

[0308] According to the method of step B2 in Example 13, cyclohexylcarboxylic acid and intermediate B-I were used as raw materials to undergo Curtis rearrangement reaction to produce intermediate B-XXVI with a yield of 63.5%. 1 H NMR (400MHz, DMSO-d6) δ8.24(d,J=8.3Hz,1H),7.01(d,J=7.9Hz,1H),6.86(d,J =8.3Hz,1H),4.52(s,1H),4.27(dd,J=11.0,5.3Hz,1H),4.04(dd,J=11.1,3.4Hz ,1H),3.35(dd,J=9.9,6.5Hz,1H),3.18(s,1H),2.75(ddd,J=10.6,7.5,2.7Hz,1 H),2.15-1.93(m,2H),1.93-1.75(m,1H),1.74-1.60(m,5H),1.20-1.01(m,5H).

[0309] Step B31: Synthesis of Intermediate B-XXVII

[0310] According to the method of step A6 in Example 1, intermediate B-XXVI was used as a raw material to undergo a nucleophilic substitution reaction with intermediate A-II to obtain intermediate B-XXVII with a yield of 50.7%. Analytical data: ESI-MS [M+H] + (m / z):564.3.

[0311] Step B32: Synthesis of Intermediate B-XXVIII

[0312] According to the method of step A7 in Example 1, intermediate B-XXVII is used as a raw material, and a reduction reaction is carried out under the action of hydrogen and palladium on carbon to obtain intermediate B-XXVIII with a yield of 88.6%.

[0313] Step B33: Synthesis of Intermediate B-XXIX

[0314] According to the method of step A8 in Example 1, intermediate B-XXVIII is used as a raw material to undergo acylation reaction with chloroacetyl chloride to obtain intermediate B-XXIX in a yield of 90.1%.

[0315] Step B34: Synthesis of Example 38

[0316] According to the method of step A9 in Example 1, intermediate B-XXIX was used as a raw material, and after nucleophilic substitution reaction with tetrahydropyrrole, deprotection was performed under acidic conditions to obtain Example 38 with a yield of 47.0%. Analytical data: ESI-MS [M+H] + (m / z):561.6. 1 H NMR(400MHz,DMSO-d6)δ12.78(s,1H),9.04(s,1H),8.71(s,1H),8.12(s,1H),7.91(s,1H),7.41-7.37(m, 3H),7.09(d,J=8.1Hz,1H),6.21(d,J=8.5Hz,1H),4.51(s,1H),4.20(dd,J=10.1,3.8Hz,1H),3.71(t,J=9. 3Hz,1H),3.66-3.55(m,1H),3.26-3.20(m,2H),3.20(s,2H),2.59(d,J=6.4Hz,4H),2.04-1.91(m,2H),1. 81(dd,J=12.7,6.4Hz,2H),1.74(d,J=5.3Hz,4H),1.58(d,J=51.8Hz,4H),1.23(s,2H),1.18-1.05(m,4H).

[0317] Example 39: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-(2-morpholinoacetylamino)pyridin-2-yl}pyrrolidin-2-yl}methylcyclohexylcarbamate (39);

[0318] According to the method of step A9 in Example 1, intermediate B-XXIX was used as a raw material, and after nucleophilic substitution reaction with morpholine, deprotection was carried out under acidic conditions to obtain Example 39 with a yield of 45.2%. Analytical data: ESI-MS [M+H] + (m / z):577.7.

[0319] Example 40: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopropylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylcyclohexylcarbamate (40);

[0320] According to the method of step A9 in Example 1, intermediate B-XXIX was used as a raw material, and after nucleophilic substitution reaction with cyclopropylamine, deprotection was carried out under acidic conditions to obtain Example 40 with a yield of 44.1%. Analytical data: ESI-MS [M+H] + (m / z):547.6. 1 H NMR (400MHz, DMSO-d6) δ12.78(s,1H),9.04(s,1H),8.71(s,1H),8.13(s,1H),7.91(s,1H),7.36(dd,J=17.9,6.6Hz,3H) ,7.10(d,J=8.0Hz,1H),6.20(d,J=8.4Hz,1H),4.55-4.50(m,1H),4.22(dd,J=10.3,3.9Hz,1H),3.72(d,J=9.6Hz,1H),3. 65(dt,J=13.0,6.7Hz,1H),3.29(s,1H),3.22-3.18(m,2H),2.18(dq,J=6.7,3.4Hz,1H),2.06-1.90(m,2H),1.76(d,J=1 5.3Hz, 4H), 1.59 (dd, J=51.0, 11.7Hz, 4H), 1.22 (d, J=10.6Hz, 2H), 1.17-1.02 (m, 4H), 0.34 (dp, J=36.0, 4.0, 3.4Hz, 4H). 13C NMR (101MHz, DMSO) δ171.40,155.90,152.81,152.45,138.00,135.61,133.47,123.67,120.92,110.70,110.36,106. 84,100.25,65.45,57.26,55.38,52.75,50.56,49.97,33.18,33.06,30.86,28.72,25.64,25.17,24.47,6.65,6.55.

[0321] Example 41: (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[(cyclopropylmethyl)amino]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}methylcyclohexylcarbamate (41);

[0322] According to the method of step A9 in Example 1, intermediate B-XXIX was used as a raw material, and after nucleophilic substitution reaction with cyclopropylmethylamine, deprotection was performed under acidic conditions to obtain Example 41 with a yield of 39.8%. Analytical data: ESI-MS [M+H] + (m / z):561.6. 1 H NMR (400MHz, DMSO-d6) δ12.65(s,1H),9.03(s,1H),8.58(s,1H),8.00(t,J=1.3Hz,1H),7.78(s,1H),7.32-7.21(m ,3H),6.96(d,J=8.1Hz,1H),6.09(d,J=8.3Hz,1H),4.40(s,1H),4.09(dd,J=10.2,3.9Hz,1H),3.63-3.50(m,1H),3 .17(s,1H),3.13(d,J=2.5Hz,1H),3.10-3.00(m,2H),2.34-2.22(m,2H),1.86(ddt,J=27.1,15.5,8.2Hz,2H),1.7 1-1.56(m,4H),1.54-1.30(m,4H),1.09(d,J=6.9Hz,1H),1.06-0.94(m,4H),0.85-0.68(m,2H),0.34-0.20(m,2H). 13C NMR(151MHz,DMSO-d6)δ171.10,155.88,152.64,152.21,137.41,135.61(2C),133.45,123.68,120.89,111.04,110.37,106.78,100.4 0,65.49,57.29,54.53(2C),52.62,50.76,49.96,33.18,33.05,28.78,25.64(2C),25.17(2C),24.52,11.48(3C),3.70(4C),3.64(4C).

[0323] Example 42: (S)-N-{{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-methylpiperazin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl}-4-(dimethylamino)benzamide (42);

[0324] The synthetic route is as follows:

[0325]

[0326] Step C1: Synthesis of Intermediate C-Ⅰ

[0327] Dissolve B-1 (3.0 g, 11.67 mmol) in THF (30 mL), add triphenylphosphine (3.67 g, 14.00 mmol) and phthalimide (2.06 g, 14.00 mmol), add diethyl azodicarboxylate (2.44 g, 14.00 mmol) dropwise at low temperature, stir at room temperature for 24 h, evaporate the reaction solution to dryness, and purify the crude product by column chromatography to obtain 3.2 g of light yellow solid, with a yield of 71%.

[0328] Step C2: Synthesis of intermediate C-II

[0329] Dissolve C-Ⅰ (3.0 g, 7.77 mmol) in ethanol (30 mL), add hydrazine hydrate (3 mL), and reflux for 2 h. Evaporate the reaction solution to dryness, and slurry the crude product with dichloromethane (15 mL). Filter and dry to obtain 1.2 g of a yellow solid with a yield of 60.3%.

[0330] Step C3: Synthesis of intermediate C-III

[0331] According to the method of step A5 in Example 1, intermediate C-II was used as a raw material to undergo acylation reaction with 4-dimethylaminobenzoyl chloride to obtain intermediate C-III with a yield of 87.2%.

[0332] Step C4: Synthesis of intermediate C-IV

[0333] According to the method of step A6 in Example 1, intermediate C-III was used as a raw material to undergo nucleophilic substitution reaction with intermediate A-II to obtain intermediate C-IV with a yield of 44.2%.

[0334] Step C5: Synthesis of Intermediate C-V

[0335] According to the method of step A6 in Example 1, intermediate C-IV was used as a raw material to undergo nucleophilic substitution reaction with intermediate A-II to obtain intermediate C-V with a yield of 44.2%.

[0336] Step C6: Synthesis of Intermediate C-VI

[0337] According to the method of step A7 in Example 1, intermediate C-V is used as a raw material, and a reduction reaction is carried out under the action of hydrogen and palladium on carbon to obtain intermediate C-VI with a yield of 84.6%.

[0338] Step C7: Synthesis of Intermediate C-VII

[0339] According to the method of step A8 in Example 1, intermediate C-VI was used as a raw material and subjected to acylation reaction with chloroacetyl chloride to obtain intermediate C-VII with a yield of 88.1%.

[0340] Step C8: Synthesis of Example 42

[0341] According to the method of step A9 in Example 1, intermediate C-VII was used as a raw material, and after a nucleophilic substitution reaction with N-methylpiperazine, deprotection was performed under acidic conditions to obtain Example 80 with a yield of 51.4%. Analytical data: ESI-MS [M+H] + (m / z):611.3.

[0342] Example 43: Anti-tumor and anti-fibrosis activity experiments on the compounds prepared in the above examples

[0343] 1. In vitro ROCK2 inhibitory activity

[0344] Using Promega The kinase detection kit was used to test the inhibitory effect of the above-obtained compounds on ROCK2. The test compound was prepared into a 10mM stock solution with DMSO, and gradiently diluted with reaction buffer (50mM HEPES (pH7.5), 10mM magnesium chloride hexahydrate, 100mM sodium orthovanadate, 0.01% CHAPS and 0.1% bovine serum albumin) to obtain compound solutions of different concentrations. In a white flat-bottom 96-well plate, 20μL ROCK2 enzyme solution (1nM), 10μL compound solution, 10μL reaction substrate (20μM RSK2 peptide KKRNRTLTK) and 10μL ATP solution (10μM) were added to each well in turn and incubated at room temperature for 3h. 50μL of the reaction detection solution in the kit was added to each well to terminate the reaction, and the chemiluminescent signal was detected using a Tecan multi-function microplate reader. Three replicates were set for all doses, and nonlinear fitting was performed using GraphPad Prism8 to calculate the IC 50 value.

[0345] The results of the inhibition of ROCK2 activity by the compounds disclosed in the present invention are shown in Table 1, in which "A" represents IC 50 Value < 50nM; "B" means 50nM < IC 50 Value < 100nM; "C" means 100nM < IC 50 Value < 1 μM; "D" means 1 μM < IC 50 Value <10μM.

[0346] Table 1 Results of the inhibition of ROCK2 activity by compounds

[0347] Example <![CDATA[ROCK2 IC 50 ]]> Example <![CDATA[ROCK2 IC 50 ]]> Example <![CDATA[ROCK2 IC 50 ]]> 1 A 16 A 31 C 2 B 17 B 32 C 3 B 18 A 33 A 4 A 19 B 34 A 5 A 20 A 35 A 6 A 21 A 36 A 7 B 22 A 37 B 8 A 23 A 38 A 9 A 24 A 39 B 10 B 25 B 40 A 11 A 26 A 41 A 12 A 27 A 42 A 13 A 28 A Belumosudil 0.153μM 14 A 29 B 15 B 30 A

[0348] As can be seen from the above, the compounds obtained in the present invention have a higher overall inhibitory activity on ROCK2, and most of the IC50 values ​​are below 100 nM, which is better than Belumosudil.

[0349] II. In vitro anti-tumor and fibrotic cell proliferation activity

[0350] Some of the compounds of the present invention were subjected to experiments on their anti-proliferation effects on human breast cancer cell line MDA-MB-231 and TGF-β-induced mouse embryonic fibroblast NIH / 3T3.

[0351] The cells were cultured in a 37°C incubator containing 5% CO2 and DMEM containing 10% fetal bovine serum. The culture medium was changed every 2 to 3 days. When the cells grew to 80% to 90% density, they were digested with trypsin and collected by centrifugation. The collected cells were resuspended in serum-containing culture medium and diluted to 2×10 4 / mL suspension. The cell suspension was inoculated into a 96-well cell culture plate at a density of 100μL / well and incubated overnight. The solution was changed and the test compound or solvent (DMSO) of each concentration gradient was added, and incubated in the incubator for 48 hours. After the treatment, the culture medium in the plate was discarded, washed twice with PBS, 100μL CCK-8 working solution was added to each well, incubated at 37℃ in the dark for 1.5 hours, and the OD was detected on the microplate reader. 450 nm The absorbance value of each well was measured and the CC of each compound was analyzed and calculated. 50 value.

[0352] The results of the anti-proliferation activities of some compounds of the present invention against MDA-MB-231 cells and TGF-β-NIH / 3T3 cells are shown in Table 2, in which "A" represents CC 50 Value < 1 μM; "B" means 1 μM < CC 50 Value <10μM; "C" means 10μM < CC 50 Values ​​<100 μM.

[0353] Table 2 Results of the anti-proliferation activities of some compounds against MDA-MB-231 cells and TGF-β-NIH / 3T3 cells

[0354]

[0355] It can be seen from the above that some of the compounds obtained in the present invention have anti-proliferative effects on MDA-MB-231 cells and TGF-β-induced NIH / 3T3 cells, among which compound 21 and compound 30 have the best anti-proliferative effects on the two cell lines. 50 All were below 1 μM.

[0356] 3. In vitro anti-tumor cell migration activity

[0357] The MDA-MB-231 cell scratch test was performed on some of the compounds of the present invention.

[0358] MDA-MB-231 cells in the logarithmic growth phase were inoculated in a 6-well plate at a density of 250,000 cells / well / 2mL and cultured in an incubator for 24 hours. The culture medium was discarded, each well was scratched with a 200-μL pipette tip and washed twice with 1mL PBS to remove the scratched cells. Each well was given 2mL of serum-free solution of compound 21 and Belumosudil, and the cell migration was observed by taking photos with an inverted microscope at 0h, 24h, and 48h. Migration rate (%) = (Area 0h -Area th ) / Area 0h ×100%, and at the same time, the Control group was given 2 mL of blank culture medium without serum as a control.

[0359] Compared with the control group, compound 21 significantly reduced the migration rate of MDA-MB-231 cells. At a concentration of 5 μM, the anti-migration effect of compound 21 was better than that of besudil. The above results indicate that compound 21 has a good in vitro anti-tumor migration effect.

[0360] IV. Anti-pulmonary fibrosis activity in vivo

[0361] Some of the compounds in the present invention were subjected to an animal model experiment based on bleomycin-induced C57BL / 6 mouse pulmonary fibrosis.

[0362] Forty C57BL / 6 mice (male, weighing 20-22g) aged 6-8 weeks were randomly divided into 4 groups, with 10 mice in each group. No food or water was allowed for 12 hours before modeling. All experimental mice were anesthetized by intraperitoneal injection of 0.05mg / kg of 2% sodium pentobarbital solution. After no response, they were hung vertically and their limbs were fixed. An indwelling needle was inserted into the mouse glottis to enter the trachea, and tracheal instillation was performed. 3U / kg of bleomycin sulfate solution (BLM) was instilled into the trachea of ​​each mouse to induce a pulmonary fibrosis model. Seven days after modeling, different concentrations of the drug solution were given by oral gavage once a day. After 14 days of administration, the mouse lung tissues were taken for HE staining to observe the thickness of the alveolar wall and the degree of neutrophil infiltration. At the same time, the lung tissues of healthy C57BL / 6 mice were used as negative controls. Figure 2 As shown in the figure, the lung tissue morphology of the model group was abnormal, with thickened alveolar walls and a large number of inflammatory cell infiltration. After the experimental group was administered with compound 21, the alveolar walls became thinner and the lung tissue morphology returned to normal. When compound 21 was administered at 100 mg / kg, the infiltration of inflammatory cells was significantly reduced, indicating that compound 21 improved the degree of inflammation in the lung tissue of mice with bleomycin-induced pulmonary fibrosis.

Claims

1. A (1H-indazol-5-yl)amino-2-pyridine derivative, characterized in that: The derivatives are compounds represented by the general formula (I) and pharmaceutically acceptable salts thereof. Where: L is R1 is unsubstituted or substituted by at least one identical or different R5 (C1-C6) alkyl, (C1-C6) alkoxy, (C3-C8) cycloalkyl, (C 2- C6)alkenyl, (C2-C6)alkynyl, or (CH2) p NR3R4; R5 is halogen, hydroxy, amino, cyano or carboxyl; p is independently 0, 1, 2 or 3; R3 and R4 are the same or different and are independently selected from hydrogen, (C1-C6) alkyl and (C3-C8) cycloalkyl which are unsubstituted or substituted by at least one identical or different R6; or R3 and R4 together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclic group containing at least one heteroatom, wherein the heterocyclic group is optionally substituted by 0-3 independent hydroxyl groups, (C1-C6) alkyl, (C1-C6) alkoxy or (C1-C6) alkoxycarbonyl which are unsubstituted or substituted by at least one identical or different R6; R6 is halogen, amino, hydroxy, cyano or (C3-C6) cycloalkyl; A is a 4-7 membered heterocyclic group containing 1-3 heteroatoms selected from N, O and S; X is selected from R2 is selected from (C1-C6)alkyl, (C3-C6)cycloalkyl, 4-6 membered heterocyclyl, aryl, which is unsubstituted or substituted by at least one identical or different R5; R7 is halogen, hydroxy, nitro, cyano, (C1-C6) alkyl, (C1-C6) alkoxy or (C1-C6) alkylamino.

2. The (1H-indazol-5-yl)amino-2-pyridine derivative according to claim 1, characterized in that: The derivatives are compounds represented by the general formula (I) and pharmaceutically acceptable salts thereof, wherein: L is R1 is unsubstituted or substituted by at least one identical or different R5 (C1-C6) alkyl, (C3-C8) cycloalkyl, (C2-C6) alkenyl or (CH2) p NR3R4; R5 is halogen, hydroxy, amino, cyano or carboxyl; wherein p is independently 0, 1, 2 or 3; R3 and R4 are the same or different and are independently selected from hydrogen, (C1-C6) alkyl and (C3-C8) cycloalkyl which are unsubstituted or substituted by at least one identical or different R6; or R3 and R4 together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclic group containing at least one heteroatom, wherein the heterocyclic group is optionally substituted by 0-3 independent hydroxyl groups, (C1-C6) alkyl, (C1-C6) alkoxy or (C1-C6) alkoxycarbonyl which are unsubstituted or substituted by at least one identical or different R6; R6 is halogen, amino, hydroxy, cyano or (C3-C6) cycloalkyl; A is a 4-7 membered heterocyclic group containing 1-3 heteroatoms selected from N or O; X is selected from One of; R2 is selected from (C1-C6)alkyl, (C3-C6)cycloalkyl, 4-6 membered heterocyclyl, aryl, which is unsubstituted or substituted by at least one identical or different R5; R7 is halogen, hydroxy, nitro, cyano, (C1-C6) alkyl, (C1-C6) alkoxy or (C1-C6) alkylamino.

3. The (1H-indazol-5-yl)amino-2-pyridine derivative according to claim 2, characterized in that: The derivatives are compounds represented by the general formula (I) and pharmaceutically acceptable salts thereof, wherein: L is R1 is unsubstituted or substituted by at least one identical or different R5 (C1-C4) alkyl, (C3-C6) cycloalkyl, (C2-C4) alkenyl or (CH2) p NR3R4; R5 is halogen, hydroxy, amino, cyano or carboxyl; Where p is independently 1, 2 or 3 R3 and R4 are the same or different and are independently selected from hydrogen, (C1-C4) alkyl and (C3-C6) cycloalkyl which are unsubstituted or substituted by at least one identical or different R6; or R3 and R4 together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclic group containing at least one heteroatom, wherein the heterocyclic group is optionally substituted by 0-1 independent hydroxyl groups, (C1-C3) alkyl groups, (C1-C3) alkyl groups which are unsubstituted or substituted by at least one identical or different R6, or (C1-C3) alkoxycarbonyl groups; R6 is amino, hydroxy or (C3-C6) cycloalkyl; A is a 5-6 membered heterocyclic group containing 1-2 heteroatoms selected from N or O; X is selected from One of; R2 is selected from (C1-C6) alkyl, (C3-C6) cycloalkyl, 5-6 membered heterocyclyl, aryl, which is unsubstituted or substituted by at least one identical or different R5; R7 is halogen, hydroxy, nitro, cyano, (C1-C4) alkyl or (C1-C4) alkylamino.

4. The (1H-indazol-5-yl)amino-2-pyridine derivative according to claim 3, characterized in that: The derivatives are compounds represented by the general formula (I) and pharmaceutically acceptable salts thereof, wherein: L is R1 is (C1-C4)alkyl, (C2-C4)alkenyl or CH2NR3R4; R3 and R4 are the same or different and are independently selected from hydrogen, (C1-C4) alkyl and (C3-C6) cycloalkyl which are unsubstituted or substituted by at least one identical or different R6; or R3 and R4 together with the nitrogen atom to which they are attached form a 5-6 membered heterocyclic group containing at least one heteroatom, wherein the heterocyclic group is optionally substituted by 0-1 independent hydroxyl, (C1-C3) alkyl, (C1-C3) alkyl which is unsubstituted or substituted by at least one identical or different R6, or methoxycarbonyl; R6 is hydroxy or cyclopropyl; A is X is selected from One of; R2 is selected from (C1-C6) alkyl, (C3-C6) cycloalkyl, 4-piperidinyl, aryl which is unsubstituted or substituted by at least one identical or different R7; R7 is halogen, (C1-C4) alkyl or (C1-C4) alkylamino.

5. The (1H-indazol-5-yl)amino-2-pyridine derivative according to claim 4, characterized in that: The derivatives are compounds represented by the general formula (I) and pharmaceutically acceptable salts thereof, wherein: L is R1 is methyl, tert-butyl, vinyl, X is selected from One of; A is R2 is 6. The (1H-indazol-5-yl)amino-2-pyridine derivative according to claim 5, characterized in that: The derivatives are the following compounds and pharmaceutically acceptable salts thereof, N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(pyrrolidin-1-yl)acetamide; N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-morpholinoacetamide; N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(piperidin-1-yl)acetamide; N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(cyclopentylamino)acetamide; N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-fluorobenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(4-methylpiperazin-1-yl)acetamide; N-{6-[(1H-indazol-5-yl)amino]-2-{4-[4-(dimethylamino)benzoyl]piperazin-1-yl}pyridin-3-yl}-2-(pyrrolidin-1-yl)acetamide; N-{6-[(1H-indazol-5-yl)amino]-2-{4-[4-(dimethylamino)benzoyl]piperazin-1-yl}pyridin-3-yl}-2-(dimethylamino)acetamide; N-{6-[(1H-indazol-5-yl)amino]-2-{4-[4-(dimethylamino)benzoyl]piperazin-1-yl}pyridin-3-yl}-2-(cyclopropylamino)acetamide; N-{6-[(1H-indazol-5-yl)amino]-2-{4-[4-(dimethylamino)benzoyl]piperazin-1-yl}pyridin-3-yl}-2-[(2-hydroxyethyl)amino]acetamide; N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-methylbenzoyl)piperazin-1-yl]pyridin-3-yl}-2-[(2-hydroxyethyl)amino]acetamide; N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-methylbenzoyl)piperazin-1-yl]pyridin-3-yl}-2-(4-hydroxypiperidin-1-yl)acetamide; N-{6-[(1H-indazol-5-yl)amino]-2-[4-(4-methylbenzoyl)piperazin-1-yl]pyridin-3-yl}-2-[4-(2-hydroxyethyl)piperazin-1-yl]acetamide; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-(methylsulfonamido)pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl{4-(dimethylamino)phenyl}carbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(dimethylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-methylpiperazin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[(cyclopropylmethyl)amino]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[(2-hydroxyethyl)amino]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}methyl[4-(dimethylamino)phenyl]carbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-acrylamidopyridin-2-yl}pyrrolidin-2-yl}p-tolylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-methylpiperazin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}p-tolylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopentylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}p-tolylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-hydroxypiperidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl p-tolylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-ethylpiperazin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl p-tolylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[4-(2-hydroxyethyl)piperazin-1-yl]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}p-tolylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl (4-fluorophenyl)carbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-pivalamidopyridin-2-yl}pyrrolidin-2-yl}methyl (4-fluorophenyl)carbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-hydroxypiperidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl (4-fluorophenyl)carbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[(cyclopropylmethyl)amino]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}methyl (4-fluorophenyl)carbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylpiperidin-4-ylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-(2-thiomorpholinoacetylamino)pyridin-2-yl}pyrrolidin-2-yl}methylpiperidin-4-ylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-methylpiperidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylpiperidin-4-ylcarbamate; (S)-1-{2-{{6-[(1H-indazol-5-yl)amino]-2-{2-[(piperidin-4-ylcarbamoyl)oxy]methyl}pyrrolidin-1-yl}pyridin-3-yl}amino-2-oxoethyl}piperidine-4-carboxylic acid methyl ester; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl isopropylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopropylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl isopropylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopentylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl isopropylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylcyclopropylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopropylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylcyclopropylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(pyrrolidin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylcyclohexylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-(2-morpholinoacetylamino)pyridin-2-yl}pyrrolidin-2-yl}methylcyclohexylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(cyclopropylamino)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methylcyclohexylcarbamate; (S)-{1-{6-[(1H-indazol-5-yl)amino]-3-{2-[(cyclopropylmethyl)amino]acetylamino}pyridin-2-yl}pyrrolidin-2-yl}methylcyclohexylcarbamate; (S)-N-{{1-{6-[(1H-indazol-5-yl)amino]-3-[2-(4-methylpiperazin-1-yl)acetylamino]pyridin-2-yl}pyrrolidin-2-yl}methyl}-4-(dimethylamino)benzamide.

7. The (1H-indazol-5-yl)amino-2-pyridine derivative according to any one of claims 1 to 6, characterized in that: The salt of the compound represented by the general formula (I) is a pharmaceutically acceptable salt obtained by reacting the compound with an acid, and the corresponding acid is hydrochloric acid, hydrobromic acid, sulfuric acid, lactic acid, citric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, tartaric acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid.

8. Use of the (1H-indazol-5-yl)amino-2-pyridine derivative according to claim 1, characterized in that: The compound represented by the general formula (I) and its pharmaceutically acceptable salt are used as ROCK2 inhibitors in the preparation of drugs for treating tumors or fibrotic diseases.

9. The use according to claim 8, characterized in that The tumor is selected from lung cancer, breast cancer, gastric cancer, colon cancer, hepatocellular carcinoma, oral cancer, kidney cancer, bladder cancer, ovarian cancer, cervical cancer, neuroblastoma, osteosarcoma, and fibrosarcoma; the fibrosis is selected from pulmonary fibrosis, renal fibrosis, liver fibrosis, and myocardial fibrosis.

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