CN111825675A

CN111825675A - ROCK inhibitor and preparation method and application thereof

Info

Publication number: CN111825675A
Application number: CN201910300698.1A
Authority: CN
Inventors: 刘军华; 陈永凯; 郭晓丹; 张轶涵; 曾弦; 钱丽娜; 王朝东
Original assignee: Wuhan LL Science and Technology Development Co Ltd; Wuhan QR Pharmaceuticals Co Ltd
Current assignee: Wuhan LL Science and Technology Development Co Ltd
Priority date: 2019-04-15
Filing date: 2019-04-15
Publication date: 2020-10-27
Anticipated expiration: 2039-04-15
Also published as: CN111825675B; WO2020211751A1

Abstract

A compound of formula I, racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, pharmaceutically acceptable salts or prodrugs thereof. The compound shown in the formula I has excellent ROCK inhibition activity. In addition, the compound shown in the formula I has better safety and metabolic stability. Further, the compounds of formula I have a low risk of potential cardiotoxicity. Finally, the compound shown in the formula I is simple in preparation method and easy to purify, so that the compound has a good application prospect.

Description

ROCK inhibitor and preparation method and application thereof

Technical Field

The invention relates to the field of medicines, in particular to a compound capable of inhibiting ROCK activity, a preparation method and application thereof.

Background

Idiopathic Interstitial Pulmonary Fibrosis (IPF) is a chronic and diffuse interstitial pulmonary disease with unknown cause and common interstitial pneumonia is mostly shown by histopathology and imaging. The disease condition is irreversibly developed due to the complex pathogenesis, and the early diagnosis is difficult; the survival rate of patients after diagnosis is remarkably reduced with the time, the 3-year survival rate is 50 percent, the 5-year survival rate is only 20 percent, the survival rate is lower than that of most cancers (such as leukemia, breast cancer, colon cancer, uterine tumor, kidney cancer and the like), and the cancer is called cancer which is not cancer. At present, no significant effective therapeutic agent for IPF is known. According to the results of the recent random control clinical trials, in combination with the actual clinical situation of China, the drugs such as pirfenidone and nintedanib can be used as appropriate, only IPF patients with mild to moderate pulmonary dysfunction are recommended to be treated by nintedanib, but IPF patients with severe pulmonary dysfunction can benefit from the nintedanib treatment, and the course of the drug administration needs to be further discussed.

Rho GTPase was discovered in 1985 and belongs to the Ras superfamily with 25% homology to Ras. Rho GTPase members found distributed in mammalian tissue cells are mainly Rho (A, B, C), Rac (1, 2, 3), Cdc42(Cdc42Hs/G25K, TC10, Tcl), Rho D, Rho G, Chp (1, 2), Rnd (Rho E/Rnd3, Rnd1/Rho6, Rnd2/Rho7), Rho H/TTF, Rif, Wrch1 and Rho BTB (1, 2), where Rho (A, B, C) is one of the most important members of Rho GTPase. ROCK (Rho-associated protein kinase) also called Rho kinase (Rho-associated kinase) belongs to serine/threonine protein kinase, has a molecular mass of about 160kD, and is the Rho downstream target effector molecule with the most detailed functional research at present. ROCK includes ROCK1(ROK β, p160-ROCK) and ROCK2(ROK α) subtypes. The amino acid sequence identity of the two subtypes was 65%, with a high degree of similarity in the kinase domain (92% identity). ROCK is distributed throughout the body, compared to ROCK1, which is more highly expressed in non-neural tissues (blood, small intestine, thymus, etc.), and ROCK2, which is more highly expressed in brain, heart and colon.

ROCK is involved in the development of a variety of cardiovascular and cerebrovascular diseases, including hypertension, atherosclerosis, ischemic stroke, heart disease, diabetic nephropathy, eye disease, tumors, nerve injury, radiation injury, autoimmune disease, and the like. For example, the Rho/ROCK signaling pathway is involved in nad (p) H oxidase activation, inducing oxidative stress, inducing cardiac microvascular damage and C-reactive protein-induced atherosclerotic thrombosis; high sugar can activate Rho/ROCK pathway, induce the expression of visceral adipokine and I-type procollagen in cardioblast cells, and cause the hyperproliferation of the cardioblast cells to induce diabetic cardiomyopathy; Rho/ROCK signal channel activation can regulate NF-kB signal channel, up-regulate inflammatory genes and induce the occurrence of diabetic nephropathy; the Rho/ROCK signal pathway changes the permeability of biological membranes to influence the metastasis of cancer cells; when spinal cord is damaged, Rho is activated, so that axon regeneration disorder caused by growth cone atrophy is induced, and inhibition effect of chondroitin sulfate proteoglycan on neuron growth is induced.

In addition, the Rho/ROCK signaling pathway is involved in the development and progression of fibrotic diseases. Rho/ROCK signaling pathway activation can increase ischemic myocardial fibrosis levels, and acute myocardial fibrosis rat heart tissue has significantly increased Rho and ROCK expression. Rho/ROCK signaling pathway activation induces actin phosphorylation, initiating cellular fibrosis. Both in vivo and in vitro results demonstrate that cardiopulmonary physiological and pathological damage after exposure to radiation for a period of time is associated with the involvement of the Rho/ROCK pathway in induced fibrosis. Endothelial adhesion fibronectin and focal adhesion formation, decreased endothelial cell migration, endothelial dysfunction due to ionizing radiation are associated with actin scaffold reorganization and stress fiber formation induced by Rho/ROCK signaling pathway activation.

Lung injury by IPF is primarily targeted at alveolar epithelial cells (ACEs), the death of which triggers wound healing responses including innate immune activation, vascular leakage and extravascular coagulation, fibroblast recruitment, proliferation and activation, extracellular matrix synthesis and crosslinking, alveolar collapse and epithelial regeneration. ROCK signals can fundamentally regulate the activity of these cells involved in the healing response, particularly epithelial cells, endothelial cells, and fibroblasts. The key role of ROCK in these responses further suggests the potential of ROCK inhibitors for the treatment of pulmonary fibrosis.

There are currently no drugs on the market that treat numerous conditions, including fibrosis, with the ROCK inhibitory pathway. The development of new agents requires careful optimization of the chemical and biological properties of the lead compounds. Further, the compounds must have desirable pharmacokinetic and pharmacodynamic characteristics. This laborious development process typically requires extensive experimentation. In many cases, the process of determining the optimal compound often requires the preparation of thousands of structurally similar compounds. Therefore, the development of a novel scaffold compound having ROCK1 and/or ROCK2 kinase inhibitory activity by improving ROCK kinase inhibitors has positive significance for the treatment of the above-mentioned diseases.

Disclosure of Invention

In order to solve the problems in the prior art, the present invention provides a compound represented by the following formula I, its racemate, stereoisomer, tautomer, isotopic label, nitrogen oxide, solvate, polymorph, metabolite, ester, pharmaceutically acceptable salt or prodrug:

wherein, X₁、X₂、X₃Identical or different, independently of one another, from CR₁、O、S、

N or NH; the R is₁Identical or different, independently of one another, from H, CN, halogen, OH, NR₁₀R₁₁、COOH、NO₂Or unsubstituted or optionally substituted by one, two or more R_aSubstituted with the following groups: c_1-40Alkyl radical, C_2-40Alkenyl radical, C_2-40Alkynyl, C_1-40Alkoxy radical, C_3-20Cycloalkyl, 3-20 membered heterocyclyl, COOR₁₀、COR₁₀、CONHR₁₀Or CONR₁₀R₁₁；

A is selected from the following groups:

wherein R is₂Same or different, independently from each other selected from H, C_1-40Alkyl radical, C_1-40Alkoxy, -X-R₁₂、-O-(CH₂)_y-CO₂R₁₂、C_6-20Aryl, 5-20 membered heteroaryl, - (CH)₂)_y-NR₁₃R₁₄、-O-(CH₂)_y-C_6-20Aryl, -O- (CH)₂)_y-5-20 membered heteroaryl, -O- (CH)₂)_y-C_3-20Cycloalkyl, -O- (CH)₂)_y-3-20 membered heterocyclyl, -O-C (═ O) - (CH)₂)_y-NR₁₃R₁₄、-O-(CH₂)_y-C(＝O)NR₁₃R₁₄、-O-(CH₂)_y-NR₁₃R₁₄、-O-(CH₂)_y-C(＝O)R₁₅、-O-(CH₂)_y-R₁₅、-O-(CH₂)_y-S(＝O)₂NR₁₃R₁₄、-O-(CH₂)_y-S(＝O)₂R₁₅、-NH-C(＝O)-(CH₂)_y-NR₁₃R₁₄、-NH-(CH₂)_y-NR₁₃R₁₄、-N-(CH₂)_y-C(＝O)NR₁₃R₁₄、-NH-C(＝O)-X-R₁₅or-NH-C (═ O) - (CH)₂)_y-R₁₅；

Each R₃Identical or different, independently of one another, from H, halogen, OH, NR₁₃R₁₄CN, or unsubstituted or optionally substituted by one, two or more R_bSubstituted with the following groups: c_1-40Alkyl radical, C_1-40Alkoxy, halo C_1-40Alkyl radical, C_3-20Cycloalkyl, 3-20 membered heterocyclyl, -C (═ O) -C_1-40Alkyl or-C (═ O) -halogeno-C_1-40An alkyl group;

m is an integer from 0 to 4; n is an integer from 0 to 2; y is an integer from 0 to 6;

y is selected from NR₄O, S or CR₄；

B is selected from the following groups:

wherein R is₅Selected from H, C_1-40Alkyl, halo C_1-40Alkyl radical, C_3-20Cycloalkyl, 3-20 membered heterocyclyl, C_2-40Alkenyl radical, C_2-40Alkynyl, - (CH)₂)_x-C(＝O)-NR₁₃R₁₄、-C(＝O)-(CH₂)_x-NR₁₃R₁₄、-C(＝O)-X-R₁₅、-(CH₂)_x-C(＝O)-NH-C_1-40alkyl-C_3-20Cycloalkyl, - (CH)₂)_x-C(＝O)-NH-C_1-40Alkyl-3-20 membered heterocyclyl, - (CH)₂)_x-C(＝O)-NH-C_1-40alkyl-C_6-20Aryl, - (CH)₂)_x-C(＝O)-NH-C_1-40Alkyl-5-20 membered heteroaryl or- (CH)₂)_x-C(＝O)-NH-C_1-40alkyl-O-C_1-40An alkyl group;

x is an integer of 0-6;

x is selected from covalent bond, O, NR₄Or C_1-40An alkyl group;

each R₄Identical or different, independently of one another, from H, or unsubstituted or optionally substituted by one, two or more R_cSubstituted with the following groups: c_1-40Alkyl, halo C_1-40Alkyl radical, C_3-20Cycloalkyl, 3-20 membered heterocyclyl, C_2-40Alkenyl radical, C_2-40Alkynyl, - (CH)₂)_z-C(＝O)-NH-C_1-40alkyl-C_3-20Cycloalkyl, - (CH)₂)_z-C(＝O)-NH-C_1-40Alkyl-3-20 membered heterocyclyl, - (CH)₂)_z-C(＝O)-NH-C_1-40alkyl-C_6-20Aryl, - (CH)₂)_z-C(＝O)-NH-C_1-40Alkyl-5-20 membered heteroaryl, - (CH)₂)_z-C(＝O)-NH-C_1-40alkyl-O-C_1-40Alkyl, or- (CH)₂)_z-NR₁₃R₁₄(ii) a z is an integer of 0 to 6;

R₁₂selected from unsubstituted or optionally substituted by one, two or more R_dSubstituted with the following groups: c_1-40Alkyl radical, C_3-20Cycloalkyl, 3-20 membered heterocyclyl, C_6-20Aryl radical, -C_1-40alkyl-C_6-20Aryl, 5-20 membered heteroaryl or-C_1-40Alkyl-5-20 membered heteroaryl;

R₁₀、R₁₁、R₁₃、R₁₄identical or different, independently of one another, from H, or unsubstituted or optionally substituted by one, two or more R_eSubstituted with the following groups: c_1-40Alkyl radical, C_2-40Alkenyl radical, C_2-40Alkynyl, C_1-40Alkoxy radical, C_3-20Cycloalkyl, 3-20 membered heterocyclyl, C_6-20Aryl radical, -C_1-40alkyl-C_6-20Aryl radical, -C_1-40Alkyl-5-20 membered heteroaryl or 5-20 membered heteroaryl; or, R₁₀、R₁₁Or R₁₃、R₁₄N attached thereto is unsubstituted or optionally substituted by one, two or more R_eSubstituted with the following groups: 3-20 membered heterocyclyl or 5-20 membered heteroaryl;

R₁₅identical or different, independently of one another, from H, or unsubstituted or optionally substituted by one, two or more R_fSubstituted with the following groups: c_1-40Alkyl radical, C_2-40Alkenyl radical, C_2-40Alkynyl radical、C_1-40Alkoxy radical, C_6-20Aryl, 5-20 membered heteroaryl, C_3-20Cycloalkyl or 3-20 membered heterocyclyl;

each R_aIdentical or different, independently of one another, from CN, halogen, OH, NR₁₆R₁₇、COOH、NO₂、C_1-40Alkyl, halo C_1-40Alkyl or C_1-40An alkoxy group;

each R_b、R_c、R_d、R_e、R_fIdentical or different, independently of one another, from halogen, OH, CN, NH₂、C_1-40Alkyl radical, C_1-40Alkoxy, -C_1-40alkyl-O-C_1-40Alkyl, halo C_1-40Alkyl, -C (═ O) -NR₁₆R₁₇、-C(＝O)-C_1-40Alkyl or C (═ O) -halogeno C_1-40An alkyl group;

R₁₆、R₁₇identical or different, independently of one another, from C_1-40Alkyl or halo C_1-40An alkyl group;

position (b) represents a linking site;

indicates the presence or absence of a conjugated ring; when not present, a heterocyclic ring is formed.

According to an embodiment of the present invention, X₁、X₂、X₃Identical or different, independently of one another, from CR₁N or NH; the R is₁Identical or different, independently of one another, from H, CN, halogen, OH, NR₁₀R₁₁、COOH、NO₂Or unsubstituted or optionally substituted by one, two or more R_aSubstituted with the following groups: c_1-10Alkyl radical, C_2-10Alkenyl radical, C_2-10Alkynyl, C_1-10Alkoxy radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl, COOR₁₀；

A is selected from the following groups:

wherein R is₂Same or different, independently from each other selected from H, C_1-10Alkyl radical, C_1-10Alkoxy radical, C_6-14Aryl, 5-14 membered heteroaryl, -X-R₁₂、-O-(CH₂)_y-CO₂R₁₂、-(CH₂)_y-NR₁₃R₁₄、-O-(CH₂)_y-C_6-14Aryl, -O- (CH)₂)_y-5-14 membered heteroaryl, -O- (CH)₂)_y-C_3-10Cycloalkyl, -O- (CH)₂)_y-3-10 membered heterocyclyl, -O-C (═ O) - (CH)₂)_y-NR₁₃R₁₄、-O-(CH₂)_y-C(＝O)NR₁₃R₁₄、-O-(CH₂)_y-NR₁₃R₁₄、-O-(CH₂)_y-C(＝O)R₁₅、-O-(CH₂)_y-R₁₅、-O-(CH₂)_y-S(＝O)₂NR₁₃R₁₄、-O-(CH₂)_y-S(＝O)₂R₁₅、-NH-C(＝O)-(CH₂)_y-NR₁₃R₁₄、-NH-(CH₂)_y-NR₁₃R₁₄、-N-(CH₂)_y-C(＝O)NR₁₃R₁₄、-NH-C(＝O)-X-R₁₅or-NH-C (═ O) - (CH)₂)_y-R₁₅；

Each R₃Identical or different, independently of one another, from H, halogen, OH, NH₂CN, or unsubstituted or optionally substituted by one, two or more R_bSubstituted with the following groups: c_1-10Alkyl radical, C_1-10Alkoxy, halo C_1-10Alkyl radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl, -C (═ O) -C_1-10Alkyl or-C (═ O) -halogeno-C_1-10An alkyl group;

m is an integer from 0 to 3; n is an integer from 0 to 2; y is an integer from 0 to 4;

y is selected from NR₄O, S or CR₄；

B is selected from the following groups:

wherein R is₅Selected from H, C_1-10Alkyl, halo C_1-10Alkyl radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl, C_2-10Alkenyl radical, C_2-10Alkynyl, - (CH)₂)_x-C(＝O)-NR₁₃R₁₄、-C(＝O)-(CH₂)_x-NR₁₃R₁₄、-C(＝O)-X-R₁₅、-(CH₂)_x-C(＝O)-NH-C_1-10alkyl-C_3-10Cycloalkyl, - (CH)₂)_x-C(＝O)-NH-C_1-10Alkyl-3-10 membered heterocyclic group, - (CH)₂)_x-C(＝O)-NH-C_1-10alkyl-C_6-14Aryl, - (CH)₂)_x-C(＝O)-NH-C_1-10Alkyl-5-14 membered heteroaryl or- (CH)₂)_x-C(＝O)-NH-C_1-10alkyl-O-C_1-10An alkyl group; x is an integer of 0-4;

x is selected from covalent bond, O, NR₄Or C_1-10An alkyl group;

each R₄Identical or different, independently of one another, from H, or unsubstituted or optionally substituted by one, two or more R_cSubstituted with the following groups: c_1-10Alkyl, halo C_1-10Alkyl radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl, C_2-10Alkenyl radical, C_2-10Alkynyl, - (CH)₂)_z-C(＝O)-NH-C_1-10alkyl-C_3-10Cycloalkyl, - (CH)₂)_z-C(＝O)-NH-C_1-10Alkyl-3-10 membered heterocyclic group, - (CH)₂)_z-C(＝O)-NH-C_1-10alkyl-C_6-14Aryl, - (CH)₂)_z-C(＝O)-NH-C_1-10Alkyl-5-14 membered heteroaryl or- (CH)₂)_z-C(＝O)-NH-C_1-10alkyl-O-C_1-10An alkyl group; z is an integer of 0 to 4;

R₁₂selected from unsubstituted or optionally substituted by one, two or more R_dSubstituted with the following groups: c_1-10Alkyl radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl, C_6-14Aryl radical, -C_1-10alkyl-C_6-14Aryl radical, -C_1-10Alkyl-5-14 membered heteroaryl or 5-14 membered heteroaryl;

R₁₀、R₁₁、R₁₃、R₁₄identical or different, independently of one another, from H, or unsubstituted or optionally substituted by one, two or more R_eSubstituted with the following groups: c_1-10Alkyl radical, C_2-10Alkenyl radical, C_2-10Alkynyl, C_1-10Alkoxy radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl, C_6-14Aryl radical, -C_1-10alkyl-C_6-14Aryl radical, -C_1-10Alkyl-5-14 membered heteroaryl or 5-14 membered heteroaryl; or, R₁₀、R₁₁Or R₁₃、R₁₄N attached thereto is unsubstituted or optionally substituted by one, two or more R_eSubstituted with the following groups: 3-10 membered heterocyclyl or 5-14 membered heteroaryl;

R₁₅identical or different, independently of one another, from H, or unsubstituted or optionally substituted by one, two or more R_fSubstituted with the following groups: c_1-10Alkyl radical, C_2-10Alkenyl radical, C_2-10Alkynyl, C_1-10Alkoxy radical, C_6-14Aryl, 5-14 membered heteroaryl, C_3-10Cycloalkyl or 3-10 membered heterocyclyl;

each R_aIdentical or different, independently of one another, from CN, halogen, OH, NR₁₆R₁₇、COOH、NO₂、C_1-10Alkyl, halo C_1-10Alkyl or C_1-10An alkoxy group;

each R_b、R_c、R_d、R_e、R_fIdentical or different, independently of one another, from halogen, OH, CN, NH₂、C_1-10Alkyl radical, C_1-10Alkoxy, -C_1-10alkyl-O-C_1-10Alkyl, halo C_1-10Alkyl, -C (═ O) -NR₁₆R₁₇、-C(＝O)-C_1-10Alkyl or-C (═ O) -halogeno-C_1-10An alkyl group; each R₁₆、R₁₇Identical or different, independently of one another, from C_1-10Alkyl or halo C_1-10An alkyl group.

According to a preferred embodiment of the present invention, wherein X₁、X₂、X₃Identical or different, independently of one another, from CR₁Or N; the R is₁Identical or different, independently of one another, from H, halogen, CN, OH, NR₁₀R₁₁、COOH、NO₂Or unsubstituted or optionally substituted by one, two or more R_aSubstituted with the following groups: c_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkyl radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl or COOR₁₀；

A is selected from the following groups:

wherein R is₂Same or different, independently from each other selected from H, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_6-14Aryl, 5-14 membered heteroaryl, -X-R₁₂、-O-(CH₂)_y-CO₂R₁₂、-(CH₂)_y-NR₁₃R₁₄、-O-(CH₂)_y-C_6-14Aryl, -O- (CH)₂)_y-5-14 membered heteroaryl, -O- (CH)₂)_y-C_3-10Cycloalkyl, -O- (CH)₂)_y-3-10 membered heterocyclyl, -O-C (═ O) - (CH)₂)_y-NR₁₃R₁₄、-O-(CH₂)_y-C(＝O)NR₁₃R₁₄、-O-(CH₂)_y-NR₁₃R₁₄、-O-(CH₂)_y-C(＝O)R₁₅、-O-(CH₂)_y-R₁₅、-O-(CH₂)_y-S(＝O)₂NR₁₃R₁₄、-O-(CH₂)_y-S(＝O)₂R₁₅、-NH-C(＝O)-(CH₂)_y-NR₁₃R₁₄、-NH-(CH₂)_y-NR₁₃R₁₄、-N-(CH₂)_y-C(＝O)NR₁₃R₁₄、-NH-C(＝O)-X-R₁₅or-NH-C (═ O) (CH)₂)_y-R₁₅；

Each R₃Identical or different, independently of one another, from H, halogen, OH, NH₂CN, or unsubstituted or optionally substituted by one, two or more R_bSubstituted with the following groups: c_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkyl radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl, -C (═ O) -C_1-6Alkyl or-C (═ O) -halogeno-C_1-6An alkyl group;

m, y are identical or different and are independently selected from 0, 1,2 or 3; n is selected from 0 or 1;

y is selected from NR₄O, S, or CR₄(ii) a Wherein said R₄Same or different, independently from each other selected from H, C_1-6Alkyl, halo C_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_3-10Cycloalkyl, 3-to 10-membered heterocyclyl, - (CH)₂)_z-C(＝O)-NH-C_1-6alkyl-C_3-10Cycloalkyl, - (CH)₂)_z-C(＝O)-NH-C_1-6Alkyl-3-10 membered heterocyclic group, - (CH)₂)_z-C(＝O)-NH-C_1-6alkyl-C_6-14Aryl, - (CH)₂)_z-C(＝O)-NH-C_1-6Alkyl-5-14 membered heteroaryl or- (CH)₂)_z-C(＝O)-NH-C_1-6alkyl-O-C_1-6An alkyl group; z is selected from 0, 1,2 or 3; r₄Preferably selected from H or C_1-6An alkyl group;

b is selected from the following groups:

wherein R is₅Selected from H, C_1-6Alkyl radical, C_2-6Alkenyl radical、C_2-6Alkynyl, halo C_1-6Alkyl radical, C_3-10Cycloalkyl, 3-to 10-membered heterocyclyl, - (CH)₂)_x-C(＝O)-NR₁₃R₁₄、-C(＝O)-(CH₂)_x-NR₁₃R₁₄、-C(＝O)-X-R₁₅、-(CH₂)_x-C(＝O)-NH-C_1-6alkyl-C_3-10Cycloalkyl, - (CH)₂)_x-C(＝O)-NH-C_1-6Alkyl-3-10 membered heterocyclic group, - (CH)₂)_x-C(＝O)-NH-C_1-6alkyl-C_6-14Aryl, - (CH)₂)_x-C(＝O)-NH-C_1-6Alkyl-5-14 membered heteroaryl or- (CH)₂)_x-C(＝O)-NH-C_1-6alkyl-O-C_1-6An alkyl group;

x is selected from O or C_1-6An alkyl group; x is selected from 0, 1,2 or 3; r₁₂Selected from unsubstituted or optionally substituted by one, two or more R_dSubstituted with the following groups: c_1-6Alkyl radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl, C_6-14Aryl radical, C_1-6alkyl-C_6-14Aryl radical, C_1-6Alkyl-5-14 membered heteroaryl or 5-14 membered heteroaryl;

R₁₀、R₁₁、R₁₃、R₁₄identical or different, independently of one another, from H, or unsubstituted or optionally substituted by one, two or more halogens, CN, NH₂Substituted with the following groups: c_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_3-10Cycloalkyl, 3-10 membered heterocyclyl, halo C_1-6Alkyl radical, C_6-14Aryl radical, C_1-6alkyl-C_6-14Aryl radical, C_1-6Alkyl-5-14 membered heteroaryl or 5-14 membered heteroaryl; or, R₁₀、R₁₁Or R₁₃、R₁₄N attached thereto forms a 3-10 membered heterocyclyl or 5-14 membered heteroaryl;

R₁₅identical or different, independently of one another, from H, or unsubstituted or optionally substituted by one, two or more halogens, OH, CN, NH₂、C_1-6Alkoxy, halo C_1-6Alkyl, -C_1-6alkyl-O-C_1-6Alkyl, -C (═ O) -C_1-6Alkyl, C (═ O) -halogeno C_1-6Alkyl-substituted groups as follows: c_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_1-6Alkoxy radical, C_6-14Aryl, 5-14 membered heteroaryl, C_3-10Cycloalkyl, 3-10 membered heterocyclyl;

each R_aIdentical or different, independently of one another, from CN, halogen, OH, NR₁₆R₁₇、COOH、NO₂、C_1-6Alkyl, halo C_1-6Alkyl or C_1-6An alkoxy group;

each R_b、R_dIdentical or different, independently of one another, from halogen, OH, CN, NH₂、C_1-6Alkyl radical, C_1-6Alkoxy, -C_1-6alkyl-O-C_1-6Alkyl, halo C_1-6Alkyl, -C (═ O) -NR₁₆R₁₇、-C(＝O)-C_1-6Alkyl or-C (═ O) -halogeno-C_1-6An alkyl group; r₁₆、R₁₇Identical or different, independently of one another, from C_1-6Alkyl or halo C_1-6An alkyl group.

According to an embodiment of the invention, the compound I is selected from the following structures:

wherein each group is as defined above.

In a preferred embodiment of the invention, the compound I is selected from the following structures:

wherein each group is as defined above.

As an example, the compound of formula I is selected from the following exemplary compounds and pharmaceutically acceptable salts thereof:

the present invention also provides a process for the preparation of a compound of formula I, comprising at least one of the following schemes:

scheme 1':

a1 ') reacting compound II-1' with compound II-2 'under alkaline condition to obtain compound II-3';

a2 ') carrying out Suzuki coupling reaction on the compound II-3' and the compound II-4 'to obtain a compound II-5';

a3 ') compound II-5' deprotection to give compound I-A;

wherein, X₁、X₂、X₃、R₂、R₃、m、Y、

Having the definition as described above, L is selected from a leaving group; PG is an amino protecting group;

or scheme 2':

b1 ') reacting the compound II-1' with the compound II-2 'under alkaline conditions to obtain a compound II-3';

b2 ') carrying out Suzuki coupling reaction on the compound II-3' and the compound II-6 'to obtain a compound II-7';

b3 ') Compound II-7' and Compound L-R₂Reacting under alkaline condition to obtain a compound II-5';

b4 ') compound II-5' deprotection to give compound I-A;

wherein, X₁、X₂、X₃、R₂、R₃、m、Y、

or scheme 3':

c1 ') reacting the compound II-1' with the compound IV-1 'under alkaline conditions to obtain a compound IV-2';

c2 ') carrying out Suzuki coupling reaction on the compound IV-2 ' and the compound IV-3 to obtain a compound IV-4 ';

c3 ') carrying out Suzuki coupling reaction on the compound IV-4 ' and the compound IV-5 ' to obtain a compound I-C;

wherein, X₁、X₂、X₃、R₂、R₃、m、Y、

Has the definition as described above, X₅L is selected from a leaving group; PG is an amino protecting group;

or scheme 4':

d1 ') reacting compound II-1' with compound II-2 'under alkaline condition to obtain compound II-3';

d2 ') reacting compound II-3' with compound III-1 'under alkaline condition to obtain compound III-2';

d3 ') deprotecting compound III-2' to give compound I-B;

wherein, X₁、X₂、X₃、R₂、R₃、n、Y、

Has the definition as described above; PG is an amino protecting group;

or scheme 5':

e1 ') reacting compound II-1' with compound IV-1 'under alkaline condition to obtain compound IV-2';

e2 ') carrying out Suzuki coupling reaction on the compound IV-2 ' and the compound IV-3 to obtain a compound IV-4 ';

e3 ') reacting compound IV-4 ' with compound III-1 ' under alkaline conditions to obtain compound I-D;

wherein, X₁、X₂、X₃、R₂、R₃、n、Y、

Has the definition as described above, X₅L is selected from a leaving group; PG is an amino protecting group.

According to the preparation process of the present invention, the leaving group and the amino-protecting group may be groups commonly used in the art, for example the leaving group may be selected from halogen (e.g. I or Cl),

And the like. Group protection or deprotection steps may optionally be performed in the above schemes and may be procedures and conditions conventional in the art.

According to the preparation process of the present invention, the base used in step a1 '), b 1'), b3 '), c 1'), d1 '), d 2'), e1 ') or e 3') is an inorganic or organic base.

According to the preparation process of the present invention, b 3') may be further carried out in the presence of a catalyst selected from at least one of sodium iodide and sodium bromide.

According to the preparation process of the present invention, the coupling reaction described in steps a2 '), b2 '), c2 '), c3 ') or e2 ') is carried out in the presence of a catalyst and a baseIs carried out by using a catalyst selected from Pd (OAc)₂And ligands XPhos, palladium tetratriphenylphosphine, or Pd (dppf) Cl₂And the like.

The deprotection step in step a3 ') or b 4') may be operations and conditions conventional in the art, for example, using acid deprotection according to the preparation method of the present invention.

According to a preferred embodiment of the invention, the compounds of formula I are prepared by the following process:

scheme 1:

a1) reacting the compound II-1 with the compound II-2' under an alkaline condition to obtain a compound II-3;

a2) carrying out Suzuki coupling reaction on the compound II-3 and the compound II-4 to obtain a compound II-5;

a3) deprotecting the compound II-5 to obtain a compound II;

wherein, X₁、X₂、X₃、R₂、R₃M, Y are as defined above, L is selected from a leaving group; PG is an amino protecting group;

or scheme 2:

b1) reacting the compound II-1 with the compound II-2' under an alkaline condition to obtain a compound II-3;

b2) carrying out Suzuki coupling reaction on the compound II-3 and the compound II-6 to obtain a compound II-7;

b3) compound II-7 and compound L-R₂Reacting under alkaline condition to obtain a compound II-5;

b4) deprotecting the compound II-5 to obtain a compound II;

or scheme 3:

c1) reacting the compound II-1 with a compound IV-1' under an alkaline condition to obtain a compound IV-2;

c2) carrying out Suzuki coupling reaction on the compound IV-2 and the compound IV-3 to obtain a compound IV-4;

c3) carrying out Suzuki coupling reaction on the compound IV-4 and the compound IV-5 to obtain a compound IV;

wherein, X₁、X₂、X₃、R₂、R₃M, Y have the meanings given above, X₅L is selected from a leaving group; PG is an amino protecting group;

or scheme 4:

d1) reacting the compound II-1 with the compound II-2' under an alkaline condition to obtain a compound II-3;

d2) reacting the compound II-3 with the compound III-1 under an alkaline condition to obtain a compound III-2;

d3) deprotecting the compound III-2 to obtain a compound III;

wherein, X₁、X₂、X₃、R₂、R₃N, Y have the definitions as described above; PG is an amino protecting group;

or scheme 5:

e1) reacting the compound II-1 with a compound IV-1' under an alkaline condition to obtain a compound IV-2;

e2) carrying out Suzuki coupling reaction on the compound IV-2 and the compound IV-3 to obtain a compound IV-4;

e3) reacting the compound IV-4 with the compound III-1 under an alkaline condition to obtain a compound V;

wherein, X₁、X₂、X₃、R₂、R₃N, Y are as defined above, X₅L is selected from a leaving group; PG is an amino protecting group.

According to embodiments of the invention, the leaving group and the amino protecting group may be groups commonly used in the art, for example the leaving group may be selected from halogen (e.g. I or Cl),

According to an embodiment of the present invention, the steps a1), b1), b3), c1), d1), d2), e1) or e3) may be operations and conditions conventional in the art, wherein the base used is an inorganic base or an organic base, for example, at least one selected from Triethylamine (TEA), potassium carbonate, isopropylamine, Diisopropylethylenediamine (DIEA), Diethylamine (DEA).

According to an embodiment of the present invention, b3) may also be carried out in the presence of a catalyst, which may be a catalyst conventional in the art, such as at least one of sodium iodide, sodium bromide.

According to an embodiment of the invention, the coupling reaction described in steps a2), b2), c2), c3) or e2) may be carried out in the presence of a catalyst such as Pd (OAc) and a base, in accordance with procedures and conditions conventional in the art₂And ligands XPhos, palladium tetratriphenylphosphine, or Pd (dppf) Cl₂And the like. For example at K₃PO₄，Pd(OAc)₂And XPhos.

According to an embodiment of the present invention, the deprotection step in step a3) or b4) may be a procedure and conditions conventional in the art, such as acid deprotection, preferably using TFA, HCl solution, e.g. using HCl dioxane solution, HCl ethyl acetate solution.

It will be understood by those skilled in the art that the compound of formula I, its racemate, stereoisomer, tautomer, nitrogen oxide can be used as a raw material or an intermediate to prepare the compound of formula I, its racemate, stereoisomer, tautomer, nitrogen oxide prodrug or pharmaceutically acceptable salt. Therefore, the invention also provides the application of the compound shown in the formula I, the racemate, the stereoisomer, the tautomer and the nitrogen oxide in preparing the compound shown in the formula I, the racemate, the stereoisomer, the tautomer, the prodrug of the nitrogen oxide or pharmaceutically acceptable salt.

The invention also provides application of at least one of a compound shown as a formula I, a racemate, a stereoisomer, a tautomer, a nitrogen oxide, an isotopic marker, a solvate, a polymorph, a metabolite, an ester, a pharmaceutically acceptable salt or a prodrug thereof in preparing a medicament, wherein the medicament is an inhibitor of protein kinase.

In particular, the agents have the function of modulating Rho-kinase. The medicament can be used for preventing or treating diseases caused by high expression of one or more ROCK or over activation of ROCK, such as cardiovascular and cerebrovascular diseases, nervous system diseases, fibrosis diseases, eye diseases, tumors, arterial thrombosis diseases, radiation injury, respiratory system diseases, autoimmune diseases and the like, including atherosclerosis, acute coronary syndrome, hypertension, cerebral vasospasm, cerebral ischemia, ischemic stroke, restenosis, heart diseases, heart failure, cardiac hypertrophy, myocardial ischemia reperfusion injury, diabetes, diabetic nephropathy, cancer, neuronal degeneration (peripheral or central), nerve injury diseases, spinal cord injury, erectile dysfunction, platelet aggregation, leukocyte aggregation, glaucoma, ocular hypertension, asthma, osteoporosis, pulmonary fibrosis (such as idiopathic pulmonary fibrosis), hepatic fibrosis, liver fibrosis, cerebral ischemia, cerebral infarction, cerebral ischemia-stroke, restenosis, heart disease, cerebral ischemia-reperfusion injury, myocardial ischemia-induced cerebral ischemia, Renal fibrosis, COPD, renal dialysis (epithelial stability), glomerulosclerosis, and neuronal degenerative inflammation.

The invention also provides a pharmaceutical composition, which comprises at least one of a therapeutically effective amount of a compound shown in formula I, a racemate, a stereoisomer, a tautomer, a nitrogen oxide, an isotopic label, a solvate, a polymorph, a metabolite, an ester, a pharmaceutically acceptable salt or a prodrug thereof.

Preferably, the pharmaceutical composition may further optionally comprise pharmaceutically acceptable excipients, such as carriers, excipients. As an example, the auxiliary material may be at least one selected from the group consisting of: disintegrants, glidants, lubricants, diluents or fillers, binders, colorants.

The pharmaceutical composition of the invention has the function of regulating Rho-kinase. The pharmaceutical composition can be used for preventing or treating diseases caused by high expression of ROCK or over activation of ROCK, such as cardiovascular and cerebrovascular diseases, nervous system diseases, fibrosis diseases, eye diseases, tumors, arterial thrombosis diseases, radiation injury, respiratory system diseases, autoimmunity diseases, including atherosclerosis, acute coronary syndrome, hypertension, cerebral vasospasm, cerebral ischemia, ischemic stroke, restenosis, heart disease, heart failure, cardiac hypertrophy, myocardial ischemia reperfusion injury, diabetes, diabetic nephropathy, cancer, neuronal degeneration (peripheral or central), nerve injury diseases, spinal cord injury, erectile dysfunction, platelet aggregation, leukocyte aggregation, glaucoma, ocular hypertension, asthma, osteoporosis, pulmonary fibrosis (such as idiopathic pulmonary fibrosis), pulmonary fibrosis, and the like, Liver fibrosis, kidney fibrosis, COPD, renal dialysis (epithelial stability), glomerulosclerosis and neuronal degenerative inflammation.

The present invention also provides a method of modulating Rho-kinase function comprising administering to a subject in need thereof an effective amount of the above pharmaceutical composition.

Definition and description of terms

Unless otherwise indicated, the definitions of groups and terms described in the specification and claims of the present application, including definitions thereof as examples, exemplary definitions, preferred definitions, definitions described in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. The definitions of the groups and the structures of the compounds in such combinations and after the combination are within the scope of the present specification.

Where a range of numerical values is recited in the specification and claims herein, and where the range of numerical values is defined as an "integer," it is understood that the two endpoints of the range are recited and each integer within the range is recited. For example, "an integer of 0 to 6" should be understood to describe each integer of 0, 1,2, 3,4, 5, and 6. "more" means three or more.

The term "halogen" refers to F, Cl, Br and I. In other words, F, Cl, Br, and I may be described as "halogen" in the present specification.

The term "C_1-40Alkyl is understood to mean a straight-chain or branched, saturated monovalent hydrocarbon radical having from 1 to 40 carbon atoms, preferably C_1-10An alkyl group. "C_1-10Alkyl "is understood to preferably mean a straight-chain or branched, saturated monovalent hydrocarbon radical having 1,2, 3,4, 5,6, 7, 8, 9 or 10 carbon atoms. The alkyl group is, for example, a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, a hexyl group, an isopropyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, an isopentyl group, a 2-methylbutyl group, a 1-ethylpropyl group, a1, 2-dimethylpropyl group, a neopentyl group, a1, 1-dimethylpropyl group, a 4-methylpentyl group, a 3-methylpentyl group, a 2-ethylbutyl group, a 1-ethylbutyl group, a3, 3-dimethylbutyl group, a2, 2-dimethylbutyl group, a1, 1-dimethylbutyl group, a2, 3-dimethylbutyl group, a1, 3-dimethylbutyl group or a1, 2-dimethylbutyl group. In particular, the radicals have 1,2, 3,4, 5,6 carbon atoms ("C)_1-6Alkyl groups) such as methyl, ethyl, propyl, butyl, isopropyl, isobutyl, sec-butyl, tert-butyl, more particularly groups having 1,2 or 3 carbon atoms ("C)_1-3Alkyl groups) such as methyl, ethyl, n-propyl or isopropyl.

The term "C_2-40Alkenyl "is understood to mean a straight-chain or branched monovalent hydrocarbon radical comprising one, two or more double bonds and having from 2 to 40 carbon atoms, preferably" C_2-10Alkenyl ". "C_2-10Alkenyl "is understood to preferably mean a straight-chain or branched, monovalent hydrocarbon radical which contains one or more double bonds and has 2,3,4, 5,6, 7, 8, 9 or 10 carbon atoms, for example C_2-6Alkenyl radical, C_2-3Alkenyl, it being understood that the alkenyl includesIn the case of more than one double bond, the double bonds may be separated from each other or conjugated. The alkenyl group is, for example, vinyl, allyl, (E) -2-methylvinyl, (Z) -2-methylvinyl, (E) -but-2-enyl, (Z) -but-2-enyl, (E) -but-1-enyl, (Z) -but-1-enyl, pent-4-enyl, (E) -pent-3-enyl, (Z) -pent-3-enyl, (E) -pent-2-enyl, (Z) -pent-2-enyl, (E) -pent-1-enyl, (Z) -pent-1-enyl, hex-5-enyl, (E) -hex-4-enyl, (Z) -hex-4-enyl, m-n-2-enyl, m-n-1-enyl, m-n-E-4-enyl, m-n-2-, (E) -hex-3-enyl, (Z) -hex-3-enyl, (E) -hex-2-enyl, (Z) -hex-2-enyl, (E) -hex-1-enyl, (Z) -hex-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E) -1-methylprop-1-enyl, (Z) -1-methylprop-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methylbut-2-enyl, (E) -2-methylbut-2-enyl, (Z) -2-methylbut-2-enyl, (E) -1-methylbut-2-enyl, (Z) -1-methylbut-2-enyl, (E) -3-methylbut-1-enyl, (Z) -3-methylbut-1-enyl, (E) -2-methylbut-1-enyl, (Z) -2-methylbut-1-enyl, (E) -1-methylbut-1-enyl, (Z) -1-methylbut-1-enyl, 1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl group and 1-isopropylvinyl group.

The term "C_2-40Alkynyl "is understood to mean a straight-chain or branched monovalent hydrocarbon radical comprising one, two or more triple bonds and having from 2 to 40 carbon atoms, preferably" C_2-10Alkynyl ". The term "C_2-10Alkynyl "is understood as preferably meaning a straight-chain or branched, monovalent hydrocarbon radical which contains one or more triple bonds and has 2,3,4, 5,6, 7, 8, 9 or 10 carbon atoms, for example C_2-3Alkynyl, C_2-6Alkynyl. The alkynyl group is, for example, ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, pent-1-ynyl, pent-2-ynyl, pent-3-ynyl, pent-4-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, hex-4-ynyl, hex-5-ynyl, 1-methylprop-2-ynyl, 2-methylbut-3-ynyl, 1-methylbut-2-ynyl, 3-methylbut-1-ynyl, 1-ethylprop-2-ynyl, prop-2-ynyl, but-3-methylbut-1-ynyl, and so-1-ethylprop, 3-methylpent-4-ynyl, 2-methylpent-4-ynyl, 1-methylpent-4-ynyl, 2-methylpent-4,2-methylpent-3-ynyl, 1-methylpent-3-ynyl, 4-methylpent-2-ynyl, 1-methylpent-2-ynyl, 4-methylpent-1-ynyl, 3-methylpent-1-ynyl, 2-ethylbut-3-ynyl, 1-ethylbut-3-ynyl, 1-ethylbut-2-ynyl, 1-propylprop-2-ynyl, 1-isopropylprop-2-ynyl, 2-dimethylbut-3-ynyl, 1-dimethylbut-2-ynyl or 3, 3-dimethylbut-1-ynyl. In particular, the alkynyl group is ethynyl, prop-1-ynyl or prop-2-ynyl.

The term "C_3-20Cycloalkyl is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 20 carbon atoms, preferably "C_3-10Cycloalkyl groups ". The term "C_3-10Cycloalkyl "is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3,4, 5,6, 7, 8, 9 or 10 carbon atoms. Said C is_3-10Cycloalkyl groups may be monocyclic hydrocarbon groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or bicyclic hydrocarbon groups such as decalin rings.

The term "3-20 membered heterocyclyl" means a saturated monovalent monocyclic or bicyclic hydrocarbon ring comprising 1-5 heteroatoms independently selected from N, O and S, preferably "3-10 membered heterocyclyl". The term "3-10 membered heterocyclyl" means a saturated monovalent monocyclic or bicyclic hydrocarbon ring comprising 1-5, preferably 1-3 heteroatoms selected from N, O and S. The heterocyclic group may be attached to the rest of the molecule through any of the carbon atoms or nitrogen atom (if present). In particular, the heterocyclic group may include, but is not limited to: 4-membered rings such as azetidinyl, oxetanyl; 5-membered rings such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6-membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, or trithianyl; or a 7-membered ring such as diazepanyl. Optionally, the heterocyclic group may be benzo-fused. The heterocyclyl group may be bicyclic, for example but not limited to a 5,5 membered ring, such as a hexahydrocyclopenta [ c ] pyrrol-2 (1H) -yl ring, or a 5,6 membered bicyclic ring, such as a hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl ring. The nitrogen atom containing ring may be partially unsaturated, i.e., it may contain one or more double bonds, such as, but not limited to, 2, 5-dihydro-1H-pyrrolyl, 4H- [1,3,4] thiadiazinyl, 4, 5-dihydrooxazolyl, or 4H- [1,4] thiazinyl, or it may be benzo-fused, such as, but not limited to, dihydroisoquinolinyl. According to the invention, the heterocyclic radical is non-aromatic.

The term "C_6-20Aryl is understood to mean a mono-, bi-or tricyclic hydrocarbon ring having a monovalent or partially aromatic character of 6 to 20 carbon atoms, preferably "C_6-14Aryl ". The term "C_6-14Aryl "is to be understood as preferably meaning a mono-, bi-or tricyclic hydrocarbon ring having a monovalent or partially aromatic character with 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms (" C_6-14Aryl group "), in particular a ring having 6 carbon atoms (" C₆Aryl "), such as phenyl; or biphenyl, or is a ring having 9 carbon atoms ("C₉Aryl), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C₁₀Aryl radicals), such as tetralinyl, dihydronaphthyl or naphthyl, or rings having 13 carbon atoms ("C₁₃Aryl radicals), such as the fluorenyl radical, or a ring having 14 carbon atoms ("C)₁₄Aryl), such as anthracenyl.

The term "5-20 membered heteroaryl" is understood to include such monovalent monocyclic, bicyclic or tricyclic aromatic ring systems: having 5 to 20 ring atoms and comprising 1 to 5 heteroatoms independently selected from N, O and S, such as "5-14 membered heteroaryl". The term "5-14 membered heteroaryl" is understood to include such monovalent monocyclic, bicyclic or tricyclic aromatic ring systems: which has 5,6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and which comprises 1 to 5, preferably 1 to 3, heteroatoms each independently selected from N, O and S and, in addition, can be benzo-fused in each case. In particular, heteroaryl is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, thia-4H-pyrazolyl and the like and their benzo derivatives, such as benzofuryl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and the like, and benzo derivatives thereof, such as quinolyl, quinazolinyl, isoquinolyl, and the like; or azocinyl, indolizinyl, purinyl and the like and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, and the like.

Unless otherwise indicated, heterocyclyl, heteroaryl or heteroarylene include all possible isomeric forms thereof, e.g., positional isomers thereof. Thus, for some illustrative, non-limiting examples, pyridyl or pyridinylene includes pyridin-2-yl, pyridinylene-2-yl, pyridin-3-yl, pyridinylene-3-yl, pyridin-4-yl, and pyridinylene-4-yl; thienyl or thienylene includes thien-2-yl, thien-3-yl and thien-3-yl.

The term "leaving group" as used herein, unless otherwise indicated, shall mean a charged or uncharged atom or group that is liberated during a substitution or displacement reaction. Suitable examples include, but are not limited to, H, F, Br, Cl, I, mesylate, tosylate, and the like.

In any of the methods for preparing the compounds of the present invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned. This can be achieved by conventional protecting groups, as described in textbooks or tool books in the art. The protecting group may be removed at a convenient subsequent stage using methods known in the art. One skilled in the art will recognize that other reagents may be used for this deprotection step, depending on the particular protecting group, including but not limited to Pd/C, Pd (OH)₂、PdCl₂、Pd(OAc)₂/Et₃SiH, raney nickel, a suitably selected acid, a suitably selected base, fluoride, and the like.

The target compound may be isolated according to known methods, for example by extraction, filtration or column chromatography.

Depending on their molecular structure, the compounds of the invention may be chiral and may therefore exist in various enantiomeric forms. These compounds may thus be present in racemic or optically active form. The compounds of the invention or intermediates thereof may be separated into enantiomeric compounds by chemical or physical methods well known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are prepared from mixtures by reaction with optically active resolving agents. Examples of suitable resolving agents are optically active acids such as the R and S forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g. N-benzoylproline or N-benzenesulfonylproline) or various optically active camphorsulphonic acids. The chromatographic enantiomeric resolution can also advantageously be carried out with the aid of optically active resolving agents, such as dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chirally derivatized methacrylate polymers, which are immobilized on silica gel. Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, for example hexane/isopropanol/acetonitrile.

Those skilled in the art will appreciate that not all nitrogen-containing heterocycles can form N-oxides, as the nitrogen needs to have available lone pairs of electrons for oxidation to an oxide; those skilled in the art will recognize nitrogen-containing heterocycles that are capable of forming N-oxides. Those skilled in the art will also recognize that tertiary amines are capable of forming N-oxides. Synthetic methods for preparing N-oxides of heterocycles and tertiary amines are well known to those skilled in the art and include the oxidation of heterocycles and tertiary amines with peroxy acids such as peracetic and m-chloroperbenzoic acid (MCPBA), hydrogen peroxide, alkyl hydroperoxides such as t-butyl hydroperoxide, sodium perborate, and dioxiranes (dioxiranes) such as dimethyldioxirane. These methods for preparing N-oxides have been widely described and reviewed in the literature.

Pharmaceutically acceptable salts may be acid addition salts of the compounds of the invention having sufficient basicity, for example having a nitrogen atom in the chain or ring, for example with the following inorganic acids: for example hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid, phosphoric acid or nitric acid, or hydrogen sulfates, or acid addition salts with organic acids such as: such as formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, caproic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2- (4-hydroxybenzoyl) benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, Mandelic acid, ascorbic acid, glucoheptylic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, hemisulfuric acid or thiocyanic acid.

In addition, another suitable pharmaceutically acceptable salt of a compound of the invention which is sufficiently acidic is an alkali metal salt (e.g., sodium or potassium salt), an alkaline earth metal salt (e.g., calcium or magnesium salt), an ammonium salt, or a salt with an organic base which affords a physiologically acceptable cation, such as a salt with: sodium ions, potassium ions, N-methylglucamine, dimethylglucamine, ethylglucamine, lysine, dicyclohexylamine, 1, 6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serinol, trihydroxymethylaminomethane, aminopropanediol, 1-amino-2, 3, 4-butanetriol. By way of example, the pharmaceutically acceptable salts include salts of the group-COOH with: sodium ion, potassium ion, calcium ion, magnesium ion, N-methylglucamine, dimethylglucamine, ethylglucamine, lysine, dicyclohexylamine, 1, 6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serinol, trihydroxymethylaminomethane, aminopropanediol, 1-amino-2, 3, 4-butanetriol.

In addition, the basic nitrogen-containing groups may be quaternized with the following agents: lower alkyl halides such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate, and diamyl sulfate; long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; aralkyl halides such as benzyl and phenethyl bromide, and the like. By way of example, pharmaceutically acceptable salts include hydrochloride, sulfate, nitrate, bisulfate, hydrobromide, acetate, oxalate, citrate, methanesulfonate, formate, or meglumine salts and the like.

Since the compound of the present invention may exist at a plurality of salt-forming sites, the "pharmaceutically acceptable salt" includes not only the salt formed at 1 salt-forming site among the compounds of the present invention but also the salt formed at 2,3 or all of the salt-forming sites among them. For this purpose, the molar ratio of the "pharmaceutically acceptable salt" of the compound of formula (I) to the cation of the acid (anion) or base required for salt formation may vary within wide limits, and may be, for example, 4:1 to 1:4, such as 3:1, 2:1, 1:2, 1:3, etc.

According to the present invention, the pharmaceutically acceptable anion includes anions selected from the group consisting of those generated by ionization of inorganic or organic acids. The "inorganic acid" includes, but is not limited to, hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid, phosphoric acid, or nitric acid. The "organic acid" includes, but is not limited to, formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, caproic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2- (4-hydroxybenzoyl) benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, maleic, Fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucoheptonic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, hemisulfuric acid or thiocyanic acid.

The term "tautomer" refers to an isomer of a functional group resulting from the rapid movement of an atom in two positions in a molecule. The compounds of the invention may exhibit tautomerism. Tautomeric compounds may exist in two or more interconvertible species. Prototropic tautomers result from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer often result in a mixture whose physicochemical properties are consistent with the mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; whereas in phenol the enol type predominates. The present invention encompasses all tautomeric forms of the compounds.

The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound of the present invention sufficient to effect the intended use, including but not limited to the treatment of a disease as defined below. The therapeutically effective amount may vary depending on the following factors: the intended application (in vitro or in vivo), or the subject and disease condition being treated, such as the weight and age of the subject, the severity of the disease condition and the mode of administration, etc., can be readily determined by one of ordinary skill in the art. The specific dosage will vary depending on the following factors: the particular compound selected, the dosage regimen to be followed, whether to administer it in combination with other compounds, the timing of administration, the tissue to be administered and the physical delivery system carried.

The term "adjuvant" refers to a pharmaceutically acceptable inert ingredient. Examples of types of excipients include, without limitation, binders, disintegrants, lubricants, glidants, stabilizers, fillers, diluents, and the like. Excipients enhance the handling characteristics of the pharmaceutical formulation, i.e., make the formulation more amenable to direct compression by increasing flowability and/or cohesiveness. Examples of typical pharmaceutically acceptable carriers suitable for use in the above formulations are: sugars such as lactose, sucrose, mannitol, and sorbitol; starches, such as corn starch, tapioca starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and methyl cellulose; calcium phosphates such as dicalcium phosphate and tricalcium phosphate; sodium sulfate; calcium sulfate; polyvinylpyrrolidone; polyvinyl alcohol; stearic acid; alkaline earth metal stearates, such as magnesium stearate and calcium stearate; stearic acid; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil and corn oil; nonionic, cationic and anionic surfactants; a glycol polymer; fatty alcohols; and grain hydrolyzed solids and other nontoxic compatible excipients commonly used in pharmaceutical formulations, such as fillers, binders, disintegrants, buffers, preservatives, antioxidants, lubricants, colorants, and the like.

The beneficial effects of the invention include:

the compound provided by the invention has excellent ROCK inhibition activity. In addition, the compound has better safety and metabolic stability. Further, the compounds of the present invention have a low risk of potential cardiotoxicity. Finally, the compound of the invention has simple preparation method and easy purification, thereby having good application prospect.

Detailed Description

The compounds of the general formula and the preparation and use thereof according to the present invention will be described in further detail with reference to the following examples. The following examples are merely illustrative and explanatory of the present invention and should not be construed as limiting the scope of the invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.

Unless otherwise indicated, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods.

< preparation examples >

Example 1 preparation of Compound (QR001)

1.1 preparation of Compound (QR001-SM1)

Potassium carbonate (18g,126mmol) was added to a solution of isopropylamine (5g,84mmol) in dichloromethane (20mL) under ice-bath, and chloroacetyl chloride (12g,100.8mmol) was slowly added dropwise, after dropping, and the reaction was allowed to proceed at room temperature overnight. Water was added, extraction was carried out with ethyl acetate, and the organic phase was dried and concentrated under reduced pressure to obtain 9g of a white solid.

¹H NMR(DMSO-d₆,400MHz):8.04(s,1H),3.98(s,2H),3.85-3.80(m,1H),1.07(d,J＝6.4Hz,6H)。

1.2 preparation of Compound (QR001-3)

To a solution of compound QR001-SM1(3g,13.6mmol) in DMF (30mL) at room temperature were added potassium carbonate (2.8g,20.4mmol) and 3-hydroxyphenylboronic acid pinacol ester (2.8g,20.4mmol, CAS: 214360-76-6), and after the addition was completed, the reaction was carried out at 80 ℃ for 16 hours. Water (50mL) was added, the mixture was extracted with ethyl acetate, the organic phase was dried, concentrated under reduced pressure, and purified by silica gel column chromatography to give 3.9g of a white solid. LC-MS: [ M + H]⁺＝320.1。

1.3 preparation of Compound (QR001-2)

2, 4-Dichloropyrrolo [2,1-f ] [1,2,4] triazine (290mg, 1.5mmol, CAS:918538-05-3) and 1-BOC-5-aminoindazole (300mg, 1.28mmol, CAS: 129488-10-4) were dissolved in a solution of ethanol (10mL), DIEA (497mg, 3.85mmol) was added dropwise, and the mixture was stirred at room temperature overnight. After the reaction, water and ethyl acetate were added to extract, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 500mg of a crude product as a yellow solid.

LC-MS:[M+1]⁺＝385.1。

1.4 preparation of Compound (QR001-4)

Compound QR001-2(200mg, 0.52mmol), boronate QR001-3(248mg, 0.78mmol, CAS:1595290-47-3), Pd (OAc)₂(11.7mg, 0.052mmol), XPhos (25mg, 0.052mmol) and K₃PO₄(330mg, 1.56mmol) was dispersed in dioxane (10mL) and water (2 mL). Heated to 110 ℃ under nitrogen for 16 hours at reflux. After completion of the reaction, it was cooled to room temperature, water and ethyl acetate were added to extract, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated and purified by column chromatography to give 100mg of a white solid. LC-MS: [ M +1 ]]⁺＝542.1。

1.5 preparation of Compound (QR001)

QR001-4(100mg,0.185mmol) was dissolved in DCM (4mL) and TFA (2mL), stirred at room temperature for 2 hours, and the reaction mixture was concentrated under reduced pressure and then subjected to preparative high performance liquid chromatography (NH)₄CO₃Method), and freeze-drying to obtain white solid 30 mg.

HNMR(400MHz，DMSO-d₆)：13.11(br,1H),9.99(s,1H),8.38(s,1H),8.15(s,1H),7.92(d,J＝8.0Hz,1H),7.86-7.84(m,2H),7.82-7.79(m,2H),7.63(d,J＝8.8Hz,1H),7.42(t,J＝8.4Hz,1H),7.19(s,1H),7.09-7.06(m,1H),6.78-6.76(m,1H),4.52(s,2H),4.01-3.94(m,1H),1.10(d,J＝6.4Hz,6H)。LC-MS:[M+H]⁺＝442.1。

Example 2 preparation of Compounds (QR003) and (QR004)

2.1 preparation of Compounds (QR003-2) and (QR003-2A)

2, 4-Dichloropyrrole [2,1-f ] [1,2,4] triazine (1.2g,6.4mmol) was dissolved in acetonitrile (40mL), and 1-chloromethyl-4-fluoro-1, 4-diazobicyclo 2.2.2 octane bis (tetrafluoroborate) salt (Selectfluor) (CAS:140681-55-6) (2.5g,7.06mmol) was added in portions at room temperature, and after stirring at room temperature for 2 hours, the reaction was continued for 16 hours at 80 ℃. After cooling and concentration, purification by silica gel column chromatography gave 980mg of the fluoro product as a yellow solid, which was nuclear-magnetically identified as a mixture of QR003-2 and QR 003-2A.

2.2 preparation of Compound (QR003-SM2)

5-Nitroindazole (1.63g, 10mmol, CAS:5401-94-5) was dissolved in 100mL of anhydrous dichloromethane, TEA (5mL) and 2- (trimethylsilyl) ethoxymethyl chloride (SEM-Cl) (1.67g, 10mmol, CAS:76513-69-4) were added at 0 deg.C, and the mixture was stirred at room temperature for 2 hours. After concentration, the mixture was purified by silica gel column chromatography to obtain QR003-SM1 in an amount of about 2.6 g.

QR003-SM1(500mg, 1.7mmol) was dissolved in 20mL of methanol, palladium on carbon (200mg) was added, and the mixture was stirred at room temperature for 3 hours under a hydrogenation atmosphere, and after completion of the reaction, the mixture was filtered, concentrated, and purified by silica gel column chromatography to obtain QR003-SM2 (about 402 mg).

2.3 preparation of Compounds (QR003-3) and (QR003-3A)

A mixture (560mg,2.73mmol) of the compounds (QR003-2) and (QR003-2A) and the compound QR003-SM2(718mg,2.73mmol) were dissolved in 25mL of tetrahydrofuran, and diisopropylethylenediamine (1.07g,8.19mmol) was added dropwise with stirring. After the addition, the reaction was carried out at room temperature for 12 hours. After the reaction solution was concentrated, it was purified by preparative silica gel plate, and the product QR003-3A 260mg was isolated as a pale yellow solid.

¹H NMR(400MHz,CDCl₃):8.29(d,J＝1.2Hz,1H),8.11(s,1H),7.68-7.59(m,2H),7.50-7.48(m,1H),7.42-7.40(m,1H),5.80(s,2H),3.61(t,J＝8.0Hz,2H),0.95(t,J＝8.0Hz,2H),0.05(s,9H)。LC-MS:[M+H]⁺＝433.0。

And separating to obtain a product QR003-3, further purifying by preparative high performance liquid chromatography, and freeze-drying to obtain 300mg of a white solid.

¹H NMR(400MHz,CDCl₃):8.12(s,1H),8.06(s,1H),7.71-7.68(m,1H),7.57-7.55(m,1H),7.51(s,1H),6.14(s,1H),5.82(s,2H),3.63(t,J＝8.0Hz,2H),0.95(t,J＝8.0Hz,2H),0.05(s,9H)。LC-MS:[M+H]⁺＝433.1。

2.4 preparation of Compounds (QR003-4) and (QR003-4A)

Compound QR003-3(300mg,0.693mmol), Borate QR001-3(331mg, 1.04mmol), Pd (OAc)₂(16mg,0.07mmol), XPhos (33mg,0.07mmol) and potassium phosphate (441mg,2.08mmol) were added to dioxane (20mL) and water (3mL), and the reaction was heated to 100 ℃ under nitrogen for 16 hours. After cooling and concentration, purification was carried out by preparative silica gel plate to obtain QR003-4 as a yellow solid of 180 mg. LC-MS: [ M + H]⁺＝590.3。

Referring to the above preparation method, compound QR003-3A (260mg,0.6mmol), boronate QR001-3(287mg, 0.9mmol), Pd (OAc) (14mg,0.06mmol), XPhos (29mg,0.06mmol) and potassium phosphate (382mg,1.8mmol) were added to dioxane (20mL) and water (3mL), and the reaction system was heated to 100 ℃ for 16 hours under nitrogen protection. After cooling and concentration, purification was carried out by preparative silica gel plate to obtain QR003-4A as a yellow solid (120 mg). LC-MS: [ M + H]⁺＝590.3.

2.5 preparation of Compounds (QR003) and (QR004)

Compound QR003-4(150mg,0.254mmol) was dissolved in trifluoroacetic acid (2mL) and dichloromethane (4mL), and stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure, purified by preparative high performance liquid chromatography, and freeze-dried to obtain 35mg of QR003 white solid.

¹H NMR(400MHz,DMSO-d₆):13.10(br,1H),10.13(s,1H),8.34(s,1H),8.16(s,1H),7.94-7.92(m,1H),7.85-7.83(m,2H),7.77-7.76(m,1H),7.64(d,J＝8.8Hz,1H),7.42(t,J＝8.0Hz,1H),7.14(br,1H),7.10-7.07(m,1H),6.49-6.47(m,1H),4.51(s,2H),4.00-3.95(m,1H),1.10(d,J＝6.4Hz,6H)。LC-MS:[M+H]⁺＝460.1。

Referring to the above preparation method, compound QR003-4A (120mg,0.2mmol) was dissolved in trifluoroacetic acid (2mL) and dichloromethane (2mL) and stirred at room temperature for 1 hour. After the reaction solution was concentrated under reduced pressure, it was purified by preparative high performance liquid chromatography, and after freeze-drying, 33mg of QR004 white solid was obtained.

¹H NMR(400MHz,DMSO-d₆):13.10(br,1H),9.32(s,1H),8.19(s,1H),8.13(s,1H),7.91-7.89(m,1H),7.75-7.70(m,4H),7.60(d,J＝4.8Hz,1H),7.37(t,J＝8.0Hz,1H),7.05-7.03(m,1H),6.68(s,1H),4.48(s,2H),3.99-3.94(m,1H),1.09(d,J＝6.8Hz,6H)。LC-MS:[M+H]⁺＝460.1。

Example 3 preparation of Compound (QR005)

3.1 preparation of Compound (QR005-2)

Compound QR001-2(500mg,1.299mmol), 3-hydroxyphenylboronic acid (213mg,1.55mmol, CAS: 87199-18-6), K₃PO₄(550mg，2.59mmol)，Pd(OAc)₂(30mg,0.132mmol) and XPhos (61mg, 0.128mmol) were added to dioxane (20mL) and water (3mL) and stirred at 100 deg.C overnight. After completion of the reaction, the reaction mixture was concentrated under reduced pressure and purified by column chromatography to obtain 200mg of a yellow solid.

LC-MS:[M+1]⁺＝343.1。

3.2 preparation of Compound (QR005-3)

Compound QR005-2(180mg, 0.52mmol) and dimethylaminopyridine (6mg, 0.05mmol) were dissolved in dichloromethane (20mL) and Boc anhydride (170mg,0.78mmol) was added under ice bath. The reaction was stirred at room temperature for 4 hours, and the reaction was terminated. Water and DCM were added for extraction, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by column chromatography to give 250mg of a yellow solid. LC-MS: [ M +1 ]]⁺＝443.3。

3.3 preparation of Compound (QR005-4)

The compound QR005-3(230mg, 0.519mmol), 2-chloro-1-morpholinoethanone (126mg, 0.779mmol, CAS: 1440-61-5 mmol), potassium carbonate (142mg, 1.03mmol) and sodium iodide (43mg, 0.26mmol) were dispersed in DMF (20mL) and the reaction was stirred at room temperature for 14 hours. After completion of the reaction, water and ethyl acetate were added to conduct extraction, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 200mg of a yellow solid.

LC-MS:[M+1]⁺＝570.3。

3.4 preparation of Compound (QR005)

Compound QR005-4(30mg, 0.05mmol) was dissolved in dichloromethane (2mL) and trifluoroacetic acid (1mL) and stirred at room temperature for 2 h. After the reaction, the mixture was concentrated, purified by preparative high performance liquid chromatography, and lyophilized to obtain 3mg of a white solid.

¹H NMR(400MHz，CD₃OD)：8.26(s,1H),8.00(s,1H),7.84-7.81(m,2H),7.76(d,J＝8.4Hz,1H),7.58(s,1H),7.51(d,J＝8.8Hz,1H),7.28(t,J＝8.4Hz,1H),6.98-6.93(m,2H),6.65-6.63(m,1H),4.73(s,2H),3.55-3.52(m,4H),3.49-3.41(m,4H)。LC-MS:[M+H]⁺＝470.1。

Example 4 preparation of Compound (QR006)

4.1 preparation of Compound (QR006-2)

2, 4-dichloropyrrolo [2,1-f ]][1,2,4]Triazine (1.5g, 7.57mmol), p-iodoaniline (1.66g,7.57mmol) and trisEthylamine (3.22mL,22.71mmol) was dispersed in 50mL of tetrahydrofuran and reacted under nitrogen at reflux for 24 hours. Cooled to room temperature, concentrated under reduced pressure, and purified by silica gel column chromatography to give 2.3g of a product as a yellow solid. LC-MS: [ M + H]⁺＝370.9。

4.2 preparation of Compound (QR006-3)

Compound QR006-2(775mg,2.094mmol), 1-BOC-pyrazole-4-boronic acid pinacol ester (739mg,2.513mmol, CAS: 552846-17-0), Pd (dppf) Cl₂(77mg,0.1mmol) and potassium carbonate (873mg,6.282mmol) were added to tetrahydrofuran (40mL) and water (8mL) and heated to 85 ℃ under nitrogen for 12 hours. Concentrated under reduced pressure and purified by column chromatography to give 318mg of the product as a yellow solid. Under long-term heating, the protecting group Boc is removed. LC-MS: [ M + H]⁺＝311.3。

4.3 preparation of Compound (QR006)

Compound QR006-3(143mg,0.46mmol), boronate QR001-3(220mg,0.69mmol), Pd (dppf) Cl₂(40mg,0.046mmol) and potassium carbonate (320mg,2.3mmol) were added to dioxane (30mL) and water (4mL), and the reaction was heated to 100 ℃ under nitrogen for 16 hours. Concentrating the reaction solution under reduced pressure, separating and purifying with silica gel plate, and performing preparative high performance liquid chromatography (NH)₄CO₃Method) and freeze-dried to obtain 5.8mg of yellow solid.

¹H NMR(400MHz,CD₃OD):13.80(br,1H)，8.68(s,2H),8.60(s,1H)，7.90(d,J＝8.0Hz,2H),7.84-7.80(m,2H),7.79-7.77(m,2H),7.66(d,J＝8.8Hz,1H),7.48(t,J＝8.4Hz,1H),7.17(s,1H),7.09-7.06(m,1H),6.79-6.77(m,1H),4.53(s,2H),4.05-3.96(m,1H),1.10(d,J＝6.4Hz,6H)。LC-MS:[M+H]⁺＝468.1。

Example 5 preparation of Compound (QR007)

5.1 preparation of Compound (QR007-2)

Compound QR005-3(230mg, 0.519mmol), N- (2-chloroethyl) propane-2-amine hydrochloride (123mg, 0).779mmol, CAS: 6306-61-2), potassium carbonate (142mg, 1.03mmol) and sodium iodide (43mg, 0.26mmol) were dispersed in DMF (20mL) and the reaction was stirred at room temperature for 14 hours. After completion of the reaction, water and ethyl acetate were added to conduct extraction, and the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 158mg of a yellow solid. LC-MS: [ M +1 ]]⁺＝528.3。

5.2 preparation of Compound (QR007)

Compound QR007-2(26mg, 0.05mmol) was dissolved in dichloromethane (2mL) and trifluoroacetic acid (1mL) and stirred at room temperature for 2 h. After the reaction, the mixture was concentrated, purified by preparative high performance liquid chromatography, and lyophilized to obtain 3mg of a white solid.

¹H NMR(400MHz，CD₃OD)：8.31(s,1H),8.02(s,1H),7.85-7.83(m,2H),7.78(d,J＝8.3Hz,1H),7.57(s,1H),7.50(d,J＝8.6Hz,1H),7.29(t,J＝8.3Hz,1H),6.98-6.93(m,2H),6.66-6.64(m,1H),4.13(t,J＝7.1Hz,1H),2.97(t,J＝7.1Hz,1H),2.83(m,1H),1.06(d,J＝6.8Hz,6H)。LC-MS:[M+H]⁺＝428.1。

Example 6 preparation of Compound (QR008)

6.1 preparation of Compound (QR008-2)

Compound QR005-3(230mg, 0.519mmol), 2-chloro-N-cyclopropylacetamide (104mg, 0.779mmol, CAS: 19047-31-5), potassium carbonate (142mg, 1.03mmol) and sodium iodide (43mg, 0.26mmol) were dispersed in DMF (20mL) and the reaction was stirred at room temperature for 14 hours. After completion of the reaction, water and ethyl acetate were added to conduct extraction, and the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 167mg of a yellow solid.

LC-MS:[M+1]⁺＝540.3。

6.2 preparation of Compound (QR008)

Compound QR008-2(27mg, 0.05mmol) was dissolved in dichloromethane (2mL) and trifluoroacetic acid (1mL) and stirred at room temperature for 2 h. After the reaction, the mixture was concentrated, purified by preparative high performance liquid chromatography, and lyophilized to obtain 3mg of a white solid.

¹H NMR(400MHz，CD₃OD)：8.31(s,1H),8.05(s,1H),7.87-7.83(m,2H),7.72(d,J＝8.4Hz,1H),7.56(s,1H),7.54(d,J＝8.8Hz,1H),7.26(t,J＝8.4Hz,1H),6.99-6.95(m,2H),6.64-6.62(m,1H),4.42(s,2H),2.69(m,1H),0.82(m,2H)，0.57(m,2H)。[M+H]⁺＝440.3.

Example 7 preparation of Compound (QR027)

7.1 preparation of Compound (QR027-2)

To a solution of compound N-BOC-5-bromoisoindoline (1.0g, 3.35mmol, CAS: 201940-08-1) in DMSO (20mL) was added bis pinacolato borate (1.02g, 4.02mmol), potassium carbonate (0.656g, 6.7mmol) and tetrakis triphenylphosphine palladium (0.30g,0.26mmol) with stirring at room temperature. The reaction system is heated to 90 ℃ under the protection of nitrogen and reacted for 16 hours. After cooling to room temperature, water was added thereto, extraction was carried out with ethyl acetate, and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. After concentration under reduced pressure, purification was performed by silica gel column chromatography to obtain 800mg of a white solid. LC-MS: [ M-55 ]]⁺＝290.1.

7.2 preparation of Compound (QR027-3)

To a solution of compound QR027-2(0.8g, 2.30mmol) in water (20mL) was added 30% hydrogen peroxide (1mL) and sodium hydroxide (1N,1mL) with stirring in an ice bath. After stirring at room temperature for 3 hours, the reaction was terminated. Diluting with ethyl acetate, filtering, washing the organic phase with sodium bicarbonate solution, concentrating at low temperature under reduced pressure, and purifying with silica gel column chromatography to obtain white solid 500 mg. LC-MS: [ M-55 ]]⁺＝180.1。

7.3 preparation of Compound (QR027-4)

To a solution of compound QR027-3(700mg, 2.97mmol) in DMF (10mL) was added iodomethane (845mg, 5.95mmol) and potassium carbonate (822mg, 5.95mmol) with stirring at room temperature. After stirring at room temperature for 5 hours, the reaction was terminated. Water was added thereto, extraction was performed with ethyl acetate, and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. Concentrating under reduced pressure, purifying with silica gel column chromatography to obtain white solidBody 600 mg. LC-MS: [ M-55 ]]⁺＝194.1.

7.4 preparation of Compound (QR027-5)

To a solution of compound QR027-4(600mg, 2.40mmol) in methanol (10mL) was added a solution of HCl in 1,4 dioxane (5mL, 2M) with stirring at room temperature. After stirring at room temperature for 5 hours, the reaction was terminated. Concentration under reduced pressure gave 300mg of the product hydrochloride as a white solid. LC-MS: [ M + H]⁺＝150.1。

7.5 preparation of Compound (QR027-6)

To a solution of compound QR027-5(38mg, 0.253mmol) in DMF (3mL) was added compound QR001-2(98mg, 0.253mmol) and DIEA (65mg, 0.506mmol) with stirring at room temperature. After the addition, the reaction mixture was heated to 100 ℃ and reacted for 16 hours. After cooling, water was added and extraction was carried out with ethyl acetate. The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. After concentration under reduced pressure, silica gel column chromatography gave 70mg of a white solid. LC-MS (LC-100)]⁺＝498.1。

7.6 preparation of Compound (QR027)

To a solution of compound QR027-6(58mg, 0.117mmol) in methanol (10mL) was added a solution of HCl in 1, 4-dioxane (3mL, 4M) with stirring at room temperature, and the reaction was terminated after stirring at room temperature for 2 hours. By preparative high performance liquid chromatography (NH)₄CO₃Method) and freeze-dried to obtain 7mg of a white solid.

¹H NMR(400MHz，DMSO-d₆)13.00(s,1H),8.59(d,J＝5.3Hz,1H),8.15(s,1H),7.88-7.85(m,1H),7.63-7.58(m,2H),7.43(d,J＝8.4Hz,1H),7.33(d,J＝6.8Hz,1H),7.20-7.17(m,1H),7.03(s,1H),6.89-6.86(m,1H),4.84-4.80(m,4H),3.77(s,3H)。LC-MS:[M+H]⁺＝398.1.

Example 8 preparation of Compound (QR029)

8.1 preparation of Compound (QR029-2)

The compound QR005-3(230mg, 0.519mmol), 2-chloro-N- (2,2, 2-trifluoroethyl) acetamide (137mg, 0.779mmol, CAS: 170655-44-4), potassium carbonate (142mg, 1.03mmol) and sodium iodide (43mg, 0.26mmol) were dispersed in DMF (20mL) and the reaction was stirred at room temperature for 14 h. After completion of the reaction, water and ethyl acetate were added to conduct extraction, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 181mg of a yellow solid. LC-MS: [ M +1 ]]⁺＝582.4。

8.2 preparation of Compound (QR029)

Compound QR029-2(30mg, 0.05mmol) was dissolved in dichloromethane (2mL) and trifluoroacetic acid (1mL) and stirred at room temperature for 2 h. After the reaction, the mixture was concentrated, purified by preparative high performance liquid chromatography, and lyophilized to obtain 3mg of a white solid.

¹H NMR(400MHz，CD₃OD)：8.32(s,1H),8.07(s,1H),7.87-7.83(m,2H),7.76(d,J＝8.3Hz,1H),7.59(s,1H),7.55(d,J＝8.4Hz,1H),7.27(t,J＝8.2Hz,1H),6.98-6.93(m,2H),6.65-6.63(m,1H),4.41(s,2H),3.72(m,2H)。LC-MS:[M+H]⁺＝482.1.

Example 9 preparation of Compound (QR031)

9.1 preparation of Compound (QR031-2)

Compound 2, 4-dichloropyrrolo [2,1-f][1,2,4]Triazine (1.52g,8.19mmol) and compound QR003-SM2(2.15g,8.19mmol) were dissolved in 75mL tetrahydrofuran and diisopropylethylenediamine (3.21g,24.57mmol) was added dropwise with stirring. After the addition, the reaction was carried out at room temperature for 12 hours. After the reaction solution is concentrated, the reaction solution is purified by silica gel column chromatography, and the product QR 031-2678 mg is obtained by separation and is light yellow solid. LC-MS: [ M + H]⁺＝415.1。

9.2 preparation of Compound (QR031-3)

Compound QR031-2(538mg,1.299mmol), 3-aminophenylborate (239mg,1.55mmol, CAS: 206658-89-1), K₃PO₄(550mg，2.59mmol)，Pd(OAc)₂(30mg,0.132mmol) and XPhos (61mg, 0.128mmol) were added to dioxane (20mL) and water (3mL) and stirred at 100 deg.C overnight. After the reaction, the reaction solution was concentrated under reduced pressure and purified by silica gel column chromatography to obtain yellow245mg of a colored solid. LC-MS: [ M +1 ]]⁺＝472.1。

9.3 preparation of Compound (QR031-4)

Compound QR031-3(200mg,0.43mmol) is added to 5mL of anhydrous dichloromethane, followed by (dimethylamino) acetyl chloride (56mg, 0.47mmol, CAS: 51552-16-0), TEA (130mg, 1.29mmol) is added dropwise, reacted at room temperature for 1 hour, after completion of the reaction, quenched with water, extracted with dichloromethane, and purified by silica gel column chromatography to give 143mg of a yellow solid. LC-MS: [ M +1 ]]⁺＝457.3。

9.4 preparation of Compound (QR031)

Compound QR031-4(23mg, 0.05mmol) is dissolved in dichloromethane (2mL) and trifluoroacetic acid (1mL) and stirred at room temperature for 2 h. After the reaction, the mixture was concentrated, purified by preparative high performance liquid chromatography, and lyophilized to obtain 7mg of a white solid.

H NMR(400MHz，DMSO-d₆)：13.08(br,1H),9.93(s,1H),8.23(s,1H),8.11(s,1H),7.82(d,J＝8.2Hz,1H),7.86-7.84(m,2H),7.85-7.82(m,2H),7.68(d,J＝8.1Hz,1H),7.47(t,J＝8.1Hz,1H),7.15(s,1H),7.09-7.06(m,1H),6.77-6.74(m,1H),4.15(s,2H),2.88(s,6H)。LC-MS:[M+H]⁺＝427.4。

Example 10

With reference to the preparation of the above examples, the following compounds were prepared:

compound numbering	LC-MS:[M+H]⁺	Compound numbering	LC-MS:[M+H]⁺
				QR002	443.2	QR021	456.2
QR009	486.3	QR022	471.2
				QR010	456.1	QR023	467.1
QR011	467.4	QR024	486.2
				QR012	472.2	QR025	510.6
QR013	467.3	QR026	478.1
				QR014	510.4	QR028	470.4
QR015	456.1	QR030	466.1
				QR016	472.2	QR032	414.5
QR017	467.4	QR033	469.3
				QR018	456.4	QR034	413.3
QR019	459.2	QR035	453.1
				QR020	510.3	QR036	549.2

。

< test for biological Activity >

1. In vitro kinase Activity evaluation

And (3) carrying out ROCK2 activity screening. ROCK2 activity was detected using a 96-well (Cisbio) time-resolved fluorometry. The ROCK2 assay was run in the following assay buffer: 5mM MgCl₂(Sigma), 1mM DTT (Sigma) and 1 Xkinase buffer. Diluting with kinase buffer, adding 3 μ L of 0.4ng/μ LROCK2 kinase (Invitrogen, PV3759) into 96 micro-well plate, adding 4 μ L of diluted test compound with DMSO content of 2.5% (volume fraction) at room temperatureIncubate for 0.5 h. To start the reaction, 6.739. mu.M ATP (alatin) and 1. mu.M of the substrate STK-substrate 2-biotin were mixed together using kinase buffer, 3. mu.L of the mixture was added to the plate, and incubation at room temperature was continued for 2 h. The reaction was stopped by mixing 5mL of detection buffer with STK antibody-Cryptate, then mixing the appropriate volume with an equal volume of streptavidin-XL 665, and adding 10. mu.L of this mixture to the microplate. After further incubation for about 1h, the plates were read on a Molecular Devices Spectra Max i3x multifunctional microplate reader. Wherein the kinase buffer, STK-substrate 2-biotin, detection buffer, STK antibody-Cryptate, and streptavidin-XL 665 are all from HTRF KinEASE-STK kit (Cisbio, 1000tests, 61 GSTMALA).

2. In vitro ROCK1 kinase selectivity assessment

ROCK1 activity was detected using a 96-well (Cisbio) time-resolved fluorometry. The ROCK1 assay was run in the following assay buffer: 5mM MgCl₂(Sigma), 1mM DTT (Sigma) and 1 Xkinase buffer. Diluting with kinase buffer solution, adding 3 μ L of 0.4ng/μ L ROCK1 kinase (Invitrogen) into 96 micro-well plate, adding 4 μ L of test compound diluted appropriately, with DMSO content of 2.5%, and incubating at room temperature for 0.5 h. To start the reaction, 3.528. mu.M ATP (alatin) and 1. mu.M of the substrate STK-substrate 2-biotin were mixed together using kinase buffer, 3. mu.L of the mixture was added to the plate and incubation was continued at room temperature for 2 h. 5mL of detection buffer was mixed with STK antibody-Cryptate, then the appropriate volume was mixed with an equal volume of streptavidin-XL 665, and 10. mu.L of the mixture was added to stop the reaction. After further incubation for about 1h, the plates were read on a molecular devices SpectraMax i3x multifunctional microplate reader. Wherein the kinase buffer, STK-substrate 2-biotin, detection buffer, STK antibody-Cryptate, and streptavidin-XL 665 are all from HTRF KinEASE-STK kit (Cisbio, 1000tests, 61 GSTMALA).

Example 11

Each of the compounds in Table 1 below was tested in ROCK2 biochemical assays by cellular assays and found to be active, in which assay the IC of all compounds₅₀All below 10. mu.M, with some compounds having IC' s₅₀Less than or equal to 100nM (active level A)Some of the ICs₅₀> 100nM and ≤ 400nM (activity level B), some IC₅₀> 400nM and ≤ 800nM (activity level C), and other IC₅₀> 800nM and ≤ 10 μ M (activity level D). Determination of ROCK2 inhibition, IC, by various Compounds₅₀The values are reported in table 1.

Inhibition of ROCK2 by the compounds of table 1

Compound numbering	IC₅₀	Compound numbering	IC₅₀
				QR001	B	QR019	C
QR002	B	QR020	B
				QR003	B	QR021	C
QR004	B	QR022	C
				QR005	B	QR023	C
QR006	C	QR024	C
				QR007	B	QR025	C
QR008	B	QR026	B
				QR009	B	QR027	C
QR010	B	QR028	B
				QR011	B	QR029	B
QR012	B	QR030	B
				QR013	B	QR031	B
QR014	B	QR032	B
				QR015	B	QR033	B
QR016	B	QR034	C
				QR017	B	QR035	C
QR018	B	QR036	B

Table 1 can determine that the compounds of the invention all have better inhibitory activity on ROCK2 kinase, and indicate the possibility of the mother nucleus fragment of the invention as ROCK2 inhibitor. Differential inhibition of ROCK1 and ROCK2 was also observed for several compounds, as shown in table 2. Table 2 below shows the different sites based on the parent nucleus structure of the compounds of the inventionSelective inhibition of ROCK1 and ROCK2 by modified compounds. IC of each compound₅₀Values (in nM) show selectivity for inhibition of ROCK 2.

Table 2: inhibition of ROCK1 and ROCK2 by compounds of the invention

EXAMPLE 12 in vitro cytotoxicity assays of Compounds of the invention

In vitro cytotoxicity assays for compounds of the invention were performed in HepG2 cells using the CCK-8 method. Collecting HepG2 cells (Beinai organisms) in logarithmic phase, adjusting the concentration of cell suspension, paving the cells in a 96-well cell culture plate by 50000cells/well, placing the cells in a cell culture box with the temperature of 37 ℃ for incubation overnight, after the cell fusion degree in the plate reaches 80-90%, adding a test compound or a solvent (DMSO) with each concentration gradient into the solution, and incubating for 48 hours in the cell culture box with the temperature of 5% and 37 ℃. After the treatment, the in-plate medium was discarded, washed with PBS 2 times, 100. mu.L of CCK-8 working solution (Biyuntian biotechnology) was added to each well, incubated at 37 ℃ for 1.5 hours in the dark, and OD was detected on an enzyme-linked immunosorbent assay_450nmAnalyzing and calculating the CC of each compound₅₀. CC obtained using the above method₅₀Shown in table 3.

TABLE 3 CC obtained for each Compound₅₀

Compound (I)	HepG2 CC₅₀(μM)
		QR001	>200
QR003	>200
		QR004	>200
QR005	>200
		QR015	>200

The experiments are also carried out on other example compounds of the invention, and the compounds of the invention are found to have better safety and CC₅₀All ranges are>10 μ M, preferably CC₅₀>30 μ M, more preferably CC₅₀>50μM。

EXAMPLE 13 in vitro metabolic stability assay of Compounds of the invention

The in vitro metabolic stability of the compounds of the invention was determined by various species of liver microsome incubation. In a liver microsome reaction System (1mg/mL liver microsome protein, 25U/mL 6-glucose phosphate dehydrogenase, 1mM NADP, 6mM D-6-glucose phosphate, 5mM MgCl₂) Adding a proper amount of test compound, putting the test compound into a water bath kettle at 37 ℃ for incubation to start reaction, adding 100 mu L of reaction liquid into an internal standard working solution (acetonitrile solution containing 200ng/mL dexamethasone, diclofenac, tolbutamide and labetalol) containing 400 mu L of 0 ℃ precooled internal standard working solution at each time point to terminate the reaction, centrifuging the mixture for 10min at 10000g of a 4 ℃ centrifuge, and taking supernatant to perform LC-MS (liquid chromatography-mass spectrometry) for analysis and detection to obtain the in-vitro metabolism half-life of the test compound in liver microsomes of various genera. T obtained by the above method_1/2Shown in table 4.

TABLE 4 Metabolic half-lives of the Compounds in liver microsomal proteins

In view of the implementation of the present inventionThe compound of the invention has better metabolic stability in human, rat and mouse, and preferably T in human liver microsome_1/2>30min, more preferably T in human liver microsomes_1/2>And (5) 90 min. The inventor also carries out comparative experiments on KD-025 which is a ROCK inhibitor entering a clinical stage, and finds that the in vitro stability of the preferred compound is better than that of KD-025.

EXAMPLE 14 assay for the Effect of Compounds of the invention on hERG Current

The effect of the compound on hERG in the invention is determined by using a hERG channel current method for detecting the stable expression of the compound on HEK293 cells by using manual patch clamp. HEK293 cell line stably expressing hERG potassium channel is cultured in DMEM medium containing 10% fetal calf serum and 0.8mg/mL G418, the culture temperature is 37 ℃, and the carbon dioxide concentration is 5%. 6 cm cell culture dishes were seeded at a density of 2.5X 105cells per dish. The cell fusion degree does not exceed 80% in order to maintain the electrophysiological activity of the cells. Before patch clamp detection, cell dissociation enzyme trypLE is used^TMExpress cells were detached, 3 × 103 cells were plated on coverslips, cultured in 24-well plates (final volume: 500 μ L), and after 18 hours, experimental assays were performed. After the whole cell sealing is formed, the cell membrane voltage is clamped at minus 80mV, the polarity is gradually removed from minus 80mV to plus 30mV for 2.5 seconds, and then the cell membrane voltage is rapidly kept at minus 50mV for 4 seconds, so that the tail current of the hERG channel can be excited. Data were collected repeatedly every 10 seconds to observe the effect of the drug on the hERG tail current. And-50 mV is used for detecting leakage current. Experimental data were collected from EPC-10 amplifiers (HEKA) and stored in PatchMaster (HEKA) software.

Administration was started when the whole cell recorded hERG current stabilized, and each drug concentration was allowed to act for 5 minutes (or current to stabilize). The cover glass with the cells is placed in a recording bath in an inverted microscope, and the test compound and the external fluid without the test compound are applied to the cells by flowing through the recording chamber from a low concentration to a high concentration in sequence by gravity perfusion, and the fluid exchange is performed by a vacuum pump during recording. The current detected by each cell in the compound-free external fluid served as its own control. Multiple cells were tested in independent replicates. All electrophysiological experiments were performed at room temperature. The effect on hERG current when 10. mu.M of compound was obtained using the method described above is shown in Table 5.

TABLE 5 Single Point inhibition of hERG at 10. mu.M concentration of each compound

Compound (I)	hERG Single-Point inhibition Rate (10. mu.M)
		QR001	23.36±0.54％
QR003	29.4±3.83％
		QR004	38.67±0.28％
QR005	8.68±0.28％
		QR015	19.34±0.3％
KD-025	74.31±0.36％

The above experiments were also performed on other example compounds of the present invention, and it was found that the compounds of the present invention have very little risk of cardiotoxicity, preferably a hERG single-point inhibition rate (10 μ M) < 50%, more preferably a hERG single-point inhibition rate (10 μ M) < 30%, and the potential cardiotoxicity risk of the compounds of the present invention is less than KD-025.

In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.

Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.

Claims

1. A compound of formula I, racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, pharmaceutically acceptable salts or prodrugs thereof:

A is selected from the following groups:

y is selected from NR₄O, S or CR₄；

B is selected from the following groups:

x is an integer of 0-6;

x is selected from covalent bond, O, NR₄Or C_1-40An alkyl group;

R₁₅identical or different, independently of one another, from H, or unsubstituted or optionally substituted by one, two or more R_fSubstituted with the following groups: c_1-40Alkyl radical, C_2-40Alkenyl radical, C_2-40Alkynyl, C_1-40Alkoxy radical, C_6-20Aryl, 5-20 membered heteroaryl, C_3-20Cycloalkyl or 3-20 membered heterocyclyl;

position (b) represents a linking site;

2. The compound of claim 1, wherein X is₁、X₂、X₃Identical or different, independently of one another, from CR₁N or NH; the R is₁Identical or different, independently of one another, from H, CN, halogen, OH, NR₁₀R₁₁、COOH、NO₂Or unsubstituted or optionally substituted by one, two or more R_aSubstituted with the following groups: c_1-10Alkyl radical, C_2-10Alkenyl radical, C_2-10Alkynyl, C_1-10Alkoxy radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl, COOR₁₀；

A is selected from the following groups:

y is selected from NR₄O, S or CR₄；

B is selected from the following groups:

x is selected from covalent bond, O, NR₄Or C_1-10An alkyl group;

each R_b、R_c、R_d、R_e、R_fIdentical or different, independently of one another, from halogen, OH, CN, NH₂、C_1-10Alkyl radical, C_1-10Alkoxy, -C_1-10alkyl-O-C_1-10Alkyl, halo C_1-10Alkyl, -C (═ O) -NR₁₆R₁₇、-C(＝O)-C_1-10Alkyl or-C (═ O) -halogeno-C_1-10An alkyl group; each one of which isR is₁₆、R₁₇Identical or different, independently of one another, from C_1-10Alkyl or halo C_1-10An alkyl group.

3. A compound according to claim 1 or 2, wherein X is₁、X₂、X₃Identical or different, independently of one another, from CR₁Or N; the R is₁Identical or different, independently of one another, from H, halogen, CN, OH, NR₁₀R₁₁、COOH、NO₂Or unsubstituted or optionally substituted by one, two or more R_aSubstituted with the following groups: c_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkyl radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl or COOR₁₀；

A is selected from the following groups:

y is selected from NR₄O, S, or CR₄(ii) a Wherein said R₄Same or different, independently from each other selected from H, C_1-6Alkyl, halo C_1-6Alkyl radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl, C_2-6Alkenyl radical, C_2-6Alkynyl, - (CH)₂)_z-C(＝O)-NH-C_1-6alkyl-C_3-10Cycloalkyl, - (CH)₂)_z-C(＝O)-NH-C_1-6Alkyl-3-10 membered heterocyclic group, - (CH)₂)_z-C(＝O)-NH-C_1-6alkyl-C_6-14Aryl, - (CH)₂)_z-C(＝O)-NH-C_1-6Alkyl-5-14 membered heteroaryl or- (CH)₂)_z-C(＝O)-NH-C_1-6alkyl-O-C_1-6An alkyl group; z is selected from 0, 1,2 or 3; r₄Preferably selected from H or C_1-6An alkyl group;

b is selected from the following groups:

wherein R is₅Selected from H, C_1-6Alkyl, halo C_1-6Alkyl radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl, C_2-6Alkenyl radical, C_2-6Alkynyl, - (CH)₂)_x-C(＝O)-NR₁₃R₁₄、-C(＝O)-(CH₂)_x-NR₁₃R₁₄、-C(＝O)-X-R₁₅、-(CH₂)_x-C(＝O)-NH-C_1-6alkyl-C_3-10Cycloalkyl, - (CH)₂)_x-C(＝O)-NH-C_1-6Alkyl-3-10 membered heterocyclic group, - (CH)₂)_x-C(＝O)-NH-C_1-6alkyl-C_6-14Aryl, - (CH)₂)_x-C(＝O)-NH-C_1-6Alkyl-5-14 membered heteroaryl or- (CH)₂)_x-C(＝O)-NH-C_1-6alkyl-O-C_1-6An alkyl group;

R₁₅identical or different, independently of one another, from H, or unsubstituted or optionally substituted by one, two or more halogens, OH, CN, NH₂、C_1-6Alkoxy, halo C_1-6Alkyl, -C_1-6alkyl-O-C_1-6Alkyl, -C (═ O) -C_1-6Alkyl, C (═ O) -halogeno C_1-6Alkyl substitutedThe following groups: c_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_1-6Alkoxy radical, C_6-14Aryl, 5-14 membered heteroaryl, C_3-10Cycloalkyl, 3-10 membered heterocyclyl;

4. A compound according to any one of claims 1 to 3, wherein compound I is selected from the following structures:

preferably, the compound I is selected from the following structures:

wherein each group has the definition as set forth in any one of claims 1 to 3.

5. The compound of any one of claims 1 to 4, wherein the compound of formula I is selected from the following exemplary compounds and pharmaceutically acceptable salts thereof:

6. a process for the preparation of a compound according to any one of claims 1 to 5, characterized in that it comprises at least one of the following schemes: scheme 1':

a3 ') compound II-5' deprotection to give compound I-A;

or scheme 2':

b4 ') compound II-5' deprotection to give compound I-A;

or scheme 3':

or scheme 4':

d3 ') deprotecting compound III-2' to give compound I-B;

or scheme 5':

wherein, X₁、X₂、X₃、R₂、R₃、m、n、Y、

Having the definition as set forth in any of claims 1 to 5, X₅L is selected from a leaving group; PG is an amino protecting group.

7. Use of at least one compound of formula I, its racemate, stereoisomer, tautomer, nitrogen oxide, isotopic label, solvate, polymorph, metabolite, ester, pharmaceutically acceptable salt or prodrug, according to any one of claims 1 to 5, for the preparation of a medicament, which is an inhibitor of protein kinases.

8. Use according to claim 7, wherein the medicament is for the prevention or treatment of a disease caused by the high expression of one or more ROCKs or the over-activation of ROCKs;

preferably, the medicament is for the prevention or treatment of: cardiovascular and cerebrovascular diseases, neurological diseases, fibrotic diseases, ocular diseases, tumors, arterial thrombotic disorders, radiation injury, respiratory diseases, and autoimmune diseases including atherosclerosis, acute coronary syndrome, hypertension, cerebral vasospasm, cerebral ischemia, ischemic stroke, restenosis, heart disease, heart failure, cardiac hypertrophy, myocardial ischemia-reperfusion injury, diabetes, diabetic nephropathy, cancer, neuronal degeneration, neuro-traumatic diseases, spinal cord injury, erectile dysfunction, platelet aggregation, leukocyte aggregation, glaucoma, ocular hypertension, asthma, osteoporosis, pulmonary fibrosis (such as idiopathic pulmonary fibrosis), hepatic fibrosis, renal fibrosis, COPD, renal dialysis, glomerulosclerosis, and neuronal degenerative inflammation.

9. A pharmaceutical composition comprising a therapeutically effective amount of at least one compound of formula I according to any one of claims 1 to 5, its racemate, stereoisomer, tautomer, nitrogen oxide, isotopic label, solvate, polymorph, metabolite, ester, pharmaceutically acceptable salt or prodrug.

10. The pharmaceutical composition of claim 9, further comprising a pharmaceutically acceptable excipient; preferably, the adjuvant is selected from at least one of the following: disintegrants, glidants, lubricants, diluents or fillers, binders, colorants.