A compound, pharmaceutical composition and use in the manufacture of a vasodilatory medicament

Abstract

The present application belongs to the technical field of vasodilative active compounds, and relates to a compound, a pharmaceutical composition and application in preparation of vasodilative drugs. The compound is selected from WRZ-1, WRZ-2, WRZ-3, WRZ-4, WRZ-5, WRZ-6, WRZ-7, WRZ-8, WRZ-9, WRZ-13, WRZ-14, WRZ-15, WRZ-21, WRZ-22, WRZ-24, WRZ-28 and WRZ-29. The compound provided by the present application has good ROCK2 inhibitory activity and good vasodilative effect.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of vasodilative active compounds, and relates to a compound, a pharmaceutical composition and application in preparation of vasodilative drugs. BACKGROUND

[0002] The information disclosed in this Background section is only for the purpose of increasing an understanding of the general context of the present application and does not necessarily constitute an acknowledgement or any form of suggestion that this information forms part of the prior art already known to a person of ordinary skill in the art.

[0003] Hypertension has become the most common cardiovascular and cerebrovascular disease in the world, and is one of the most dangerous factors causing abnormal human death. In medicine, the systolic / diastolic blood pressure of a blood vessel under resting state ≥ 140 / 90 mmHg is taken as the standard for judging hypertension, and most people have blood pressure ≥ 140 / 90 mmHg measured twice or more in the morning, which can be preliminarily diagnosed as hypertension. Hypertension does not exist alone, and patients often have metabolic disorders of blood sugar and blood lipids, and even pathological changes of organs such as heart, kidney and brain at the same time, so hypertension is an extremely complex systemic disease. The causes of hypertension are very complex, including genetic inheritance, living environment, bad living habits and long-term medication. About 1 / 3 of the world's population has high blood pressure, and there are more than 330 million hypertensive patients in China, so it is urgent to develop new antihypertensive drugs.

[0004] Studies have confirmed that high expression of Rho-associated kinase (Rho-associated kinase, ROCK) can contract blood vessels in multiple ways, and inhibition of its activity can significantly dilate blood vessels to achieve the purpose of reducing blood pressure. ROCK belongs to the serine / threonine kinase family and is a direct and main downstream effector of GTP-bound RhoA protein, and is divided into two subtypes of ROCK1 and ROCK2. ROCK1 is mainly highly expressed in liver, spleen, kidney, lung and testis, while ROCK2 is highly expressed in brain, heart and blood vessels. Compared with ROCK1, the activation of ROCK2 is more closely related to cardiovascular system homeostasis and diseases related to vascular contraction, and is considered to be an effective target for vasodilation and treatment of hypertension. Inhibition of its high expression can effectively dilate blood vessels to achieve the purpose of treating hypertension. ROCK2 inhibitors not only can promote the secretion of endogenous vasodilative factors, but also can directly act on myosin light chain to inhibit cell contraction, thereby producing a hypotensive effect. At present, a number of ROCK inhibitors with vasodilative effect have been reported, such as fasudil, ripasudil and netarsudil. In order to design and synthesize small molecule compounds with better vasodilative effect and stronger ROCK2 inhibitory effect, and finally find a highly efficient and low-toxicity drug for treating hypertension, the present application is proposed. SUMMARY

[0005] The present application aims to provide a compound, a pharmaceutical composition and an application in preparing vasodilator drugs, and the compound provided by the present application has good ROCK2 inhibitory activity and good vasodilating effect.

[0006] In order to achieve the above-mentioned purpose, the technical scheme of the present application is:

[0007] In one aspect, a compound is selected from the following:

[0008] N-(1H-indazol-5-yl)-2-(3-methoxyphenyl)acetamide (compound code WRZ-1);

[0009] 2-(3-fluorophenyl)-N-(1H-indazol-5-yl)acetamide (compound code WRZ-2);

[0010] N-(1H-indazol-5-yl)-2-(m-tolyl)acetamide (compound code WRZ-3);

[0011] N-(1H-indazol-5-yl)-2-(3-(trifluoromethyl)phenyl)acetamide (compound code WRZ-4);

[0012] 2-(3,4-dimethoxyphenyl)-N-(1H-indazol-5-yl)acetamide (compound code WRZ-5);

[0013] 2-bromo-N-(1H-indazol-5-yl)-2-(3-methoxyphenyl)acetamide (compound code WRZ-6);

[0014] 2-bromo-2-(3-fluorophenyl)-N-(1H-indazol-5-yl)acetamide (compound code WRZ-7);

[0015] 2-bromo-N-(1H-indazol-5-yl)-2-(m-tolyl)acetamide (compound code WRZ-8);

[0016] 2-bromo-2-(3,4-dichlorophenyl)-N-(1H-indazol-5-yl)acetamide (compound code WRZ-9);

[0017] 2-(3-fluorophenyl)-N-(quinolin-6-yl)acetamide (compound code WRZ-14);

[0018] N-(quinolin-6-yl)-2-(m-tolyl)acetamide (compound code WRZ-15);

[0019] 2-bromo-2-(3-methoxyphenyl)-N-(quinolin-6-yl)acetamide (compound code WRZ-21);

[0020] 2-bromo-2-(3-fluorophenyl)-N-(quinolin-6-yl)acetamide (compound code WRZ-22);

[0021] 2-bromo-2-(3,4-dichlorophenyl)-N-(quinolin-6-yl)acetamide (compound code WRZ-24);

[0022] N-hydroxy-4-(3-(isoquinolin-5-yl)ureido)benzamide (compound code WRZ-28);

[0023] 4-(3-(isoquinolin-5-yl)ureido)benzamide (compound code WRZ-29).

[0024] In order to obtain compounds with good enzyme inhibition activity on ROCK2, three main ring structures are designed in the present application, as shown below:

[0025]

[0026] wherein, R 1 is a hydrogen atom or a bromine atom, R 2 is 3-OCH3, 3-CH3, 3-F, 3-Cl, 3-CF3, 3,4-2OCH3, 3,4-2Cl, 3,4-2F, R 3 is 4-CONH2, 3-CH3.

[0027] It has been found through research that among the compounds based on the above three main ring structures, the substituents have a great influence on the ROCK2 enzyme inhibition rate of the compounds. Among them, the inhibition rates of the above 16 compounds provided by the present application on ROCK2 enzyme are all more than 50%, and all have good ROCK2 enzyme inhibition activity, while the ROCK2 enzyme inhibition activity of other synthesized compounds is poor, for example, the inhibition rate of 2-(3-chlorophenyl)-N-(quinolin-6-yl)acetamide (compound code WRZ-18) on ROCK2 enzyme is only 0.3%.

[0028] In some embodiments, the compound is selected from the following:

[0029] N-(1H-indazol-5-yl)-2-(3-methoxyphenyl)acetamide (compound code WRZ-1);

[0030] 2-(3-fluorophenyl)-N-(1H-indazol-5-yl)acetamide (compound code WRZ-2);

[0031] N-(1H-indazol-5-yl)-2-(m-tolyl)acetamide (compound code WRZ-3);

[0032] N-(1H-indazol-5-yl)-2-(3-(trifluoromethyl)phenyl)acetamide (Compound Code: WRZ-4);

[0033] 2-bromo-N-(1H-indazol-5-yl)-2-(3-methoxyphenyl)acetamide (Compound Code: WRZ-6);

[0034] 2-bromo-2-(3-fluorophenyl)-N-(1H-indazol-5-yl)acetamide (Compound Code: WRZ-7);

[0035] 2-bromo-N-(1H-indazol-5-yl)-2-(m-tolyl)acetamide (Compound Code: WRZ-8);

[0036] 2-bromo-2-(3,4-dichlorophenyl)-N-(1H-indazol-5-yl)acetamide (Compound Code: WRZ-9);

[0037] N-(quinolin-6-yl)-2-(m-tolyl)acetamide (Compound Code: WRZ-15);

[0038] 2-bromo-2-(3-methoxyphenyl)-N-(quinolin-6-yl)acetamide (Compound Code: WRZ-21);

[0039] 2-bromo-2-(3-fluorophenyl)-N-(quinolin-6-yl)acetamide (Compound Code: WRZ-22);

[0040] 2-bromo-2-(3,4-dichlorophenyl)-N-(quinolin-6-yl)acetamide (Compound Code: WRZ-24);

[0041] N-hydroxy-4-(3-(isoquinolin-5-yl)ureido)benzamide (Compound Code: WRZ-28).

[0042] Further research shows that the above-mentioned compounds provided by the present application have an inhibition rate of more than 80% on ROCK2 enzyme, and have better ROCK2 enzyme inhibition activity.

[0043] In some embodiments, the compound is selected from the following:

[0044] N-(1H-indazol-5-yl)-2-(3-methoxyphenyl)acetamide (Compound Code: WRZ-1);

[0045] 2-(3-fluorophenyl)-N-(1H-indazol-5-yl)acetamide (Compound Code: WRZ-2);

[0046] N-(1H-indazol-5-yl)-2-(m-tolyl)acetamide (Compound Code: WRZ-3);

[0047] N-(1H-indazol-5-yl)-2-(3-(trifluoromethyl)phenyl)acetamide (Compound Code: WRZ-4);

[0048] 2-bromo-N-(1H-indazol-5-yl)-2-(3-methoxyphenyl)acetamide (Compound Code: WRZ-6);

[0049] 2-bromo-2-(3-fluorophenyl)-N-(1H-indazol-5-yl)acetamide (Compound Code: WRZ-7);

[0050] 2-bromo-N-(1H-indazol-5-yl)-2-(m-tolyl)acetamide (Compound Code: WRZ-8);

[0051] 2-bromo-2-(3,4-dichlorophenyl)-N-(1H-indazol-5-yl)acetamide (Compound Code: WRZ-9);

[0052] N-(quinolin-6-yl)-2-(m-tolyl)acetamide (Compound Code: WRZ-15);

[0053] 2-bromo-2-(3-methoxyphenyl)-N-(quinolin-6-yl)acetamide (Compound Code: WRZ-21);

[0054] 2-bromo-2-(3-fluorophenyl)-N-(quinolin-6-yl)acetamide (Compound Code: WRZ-22).

[0055] Further research shows that the above-mentioned compounds provided by the application have an inhibition rate of more than 90% on ROCK2 enzyme, and have excellent ROCK2 enzyme inhibition activity, which is equivalent to that of the positive drug Fasudil.

[0056] In some embodiments, the compound is selected from the following:

[0057] N-(1H-indazol-5-yl)-2-(3-methoxyphenyl)acetamide (Compound Code: WRZ-1);

[0058] N-(1H-indazol-5-yl)-2-(3-(trifluoromethyl)phenyl)acetamide (Compound Code: WRZ-4);

[0059] 2-bromo-N-(1H-indazol-5-yl)-2-(3-methoxyphenyl)acetamide (Compound Code: WRZ-6);

[0060] 2-bromo-2-(3-fluorophenyl)-N-(1H-indazol-5-yl)acetamide (Compound Code: WRZ-7).

[0061] The compound provided by the present application has a higher inhibition rate on ROCK2 enzyme than the positive drug Fasudil, and shows stronger ROCK2 enzyme inhibition activity, compared with the positive control drug Fasudil. Among them, 2-bromo-N-(1H-indazol-5-yl)-2-(3-methoxyphenyl)acetamide (compound code WRZ-6) has the best ROCK2 enzyme inhibition activity.

[0062] In another aspect, a compound is 2-(3-methoxyphenyl)-N-(quinolin-6-yl)acetamide (compound code WRZ-13). Studies have shown that although the inhibition rate of WRZ-13 on ROCK2 enzyme is poor, it has a good effect on relaxing vasoconstriction.

[0063] The synthesis route of the compound shown in formula I is as follows:

[0064]

[0065] The specific synthesis steps are as follows:

[0066] The benzene acetic acid (1a-1h) containing the substituent group R 2 is acylated to form an intermediate acyl chloride (4a-4h), and then reacted with 5-aminoindazole in a tetrahydrofuran solution to form the target compounds WRZ-1-WRZ-5. The benzene acetic acid (1a-1h) containing the substituent group R 2 undergoes a free radical substitution reaction under the action of NBS, replacing one hydrogen atom on the benzyl group with a bromine atom to obtain an intermediate (2a-2h), and then the benzene acetic acid is reacted with oxalyl chloride or thionyl chloride to form an acyl chloride (3a-3h), and then reacted with 5-aminoindazole in a tetrahydrofuran solution to form the target compounds WRZ-6-WRZ-12.

[0067] The synthesis route of the compound shown in formula II is as follows:

[0068]

[0069] The specific synthesis steps are as follows:

[0070] The benzene acetic acid (1a-1h) containing the substituent group R 2 is acylated to form an intermediate acyl chloride (4a-4h), and then reacted with 6-aminoquinoline in a dichloromethane solution to form the target compounds WRZ-13-WRZ-20. The benzene acetic acid (1a-1h) containing the substituent group R 2The benzene acetic acid (1a-1h) is subjected to free radical substitution reaction under the action of NBS, one hydrogen atom on the benzyl is replaced by bromine atom to obtain intermediate (2a-2h), then the benzene acetic acid is subjected to acyl chloride reaction by oxalyl chloride or thionyl chloride to obtain acyl chloride (3a-3h), then the acyl chloride is reacted with 6-aminoquinoline in dichloromethane solution to obtain the target compound.

[0071] The synthesis route of the compound shown in formula III is as follows:

[0072]

[0073] The specific synthesis steps are as follows:

[0074] The aniline (5a-5h) containing substituent R 1 is subjected to isocyanate reaction (6a-6b) by the action of triphosgene, then reacted with 5-aminoisoquinoline to obtain urea intermediate (7a-7b). The intermediate 7a is subjected to hydroxylamine hydrochloride and potassium hydroxide to obtain hydroxamic acid, and the target compound WRZ-28 is obtained; the intermediate 7b is subjected to formamide by the action of 30% hydrogen peroxide, and the target compound WRZ-29 is obtained.

[0075] Since the effect of WRZ-6 is the best in the compounds provided in the present application, the third aspect provides a preparation method of WRZ-6, that is, a preparation method of a compound, comprising the steps according to the following reaction route:

[0076]

[0077] Specifically, 2-methoxyphenyl acetic acid is subjected to free radical substitution reaction under the action of NBS, one hydrogen atom on the benzyl is replaced by bromine atom to obtain 2-bromo-2-(3-methoxyphenyl) acetic acid, 2-bromo-2-(3-methoxyphenyl) acetic acid is subjected to acyl chloride reaction by oxalyl chloride or thionyl chloride to obtain 2-bromo-2-(3-methoxyphenyl) acetyl chloride, and 2-bromo-2-(3-methoxyphenyl) acetyl chloride is subjected to amidation reaction with 5-aminoindazole to obtain.

[0078] The fourth aspect is a pharmaceutical composition comprising the above-mentioned compound, a pharmaceutically acceptable salt of the compound, a solvate of the compound, a hydrate of the compound or a stereoisomer of the compound.

[0079] The pharmaceutically acceptable salt of the present application is a derivative after group modification for improving the physical and chemical properties of the compound, which is usually a salt formed by the compound and inorganic salt such as hydrochloric acid, sulfuric acid, nitric acid or hydrobromic acid, etc., and a salt formed by the compound and organic acid, such as methanesulfonic acid, toluenesulfonic acid, citric acid or trifluoroacetic acid, etc.

[0080] The solvate according to the present application refers to a crystal substance formed with an organic solvent (ethanol, methanol, etc.).

[0081] In a fifth aspect, a pharmaceutical preparation comprises an active ingredient and a pharmaceutical carrier, wherein the active ingredient is the compound or the pharmaceutical composition described above.

[0082] The pharmaceutical carrier according to the present application includes, but is not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, buffer substances (such as phosphates), glycerol, sorbitol, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts), colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, beeswax, lanolin, etc.; the content of the pharmaceutical carrier in the pharmaceutical composition can be 1%-98% by weight, usually about 80% by weight.

[0083] Preferably, the pharmaceutical preparation can be administered by oral administration, spray inhalation, rectal administration, nasal administration, vaginal administration, topical administration, parenteral administration such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal or intracranial injection or infusion, or by means of an implanted reservoir, and more preferably by oral administration, intramuscular injection, intraperitoneal injection or intravenous injection.

[0084] Preferably, the dosage form of the pharmaceutical preparation can be a liquid dosage form or a solid dosage form; wherein the liquid dosage form can be a true solution, a colloid, a microparticle, an emulsion, a suspension; and the solid dosage form includes tablets, capsules, dripping pills, aerosols, pills, powders, emulsions, granules, suppositories, freeze-dried powder injections, clathrates, landfill agents, patches, rubs, etc.

[0085] In a sixth aspect, the compound, the pharmaceutical composition or the pharmaceutical preparation described above is used for preparing a ROCK inhibitor.

[0086] Specifically, the ROCK inhibitor can be used for preparing a model drug for a ROCK inhibition model, can be used for preventing, improving or treating a disease related to high expression of ROCK, and can be used for preparing a disease model, which can be a cell, tissue or animal model.

[0087] Specifically, the ROCK inhibitor can be used for preparing a model drug for a ROCK inhibition model, can be used for preventing, improving or treating a disease related to high expression of ROCK, and can be used for preparing a disease model, which can be a cell, tissue or animal model.

[0088] Specifically, the ROCK inhibitor is a ROCK2 inhibitor.

[0089] In a seventh aspect, use of the compound, pharmaceutical composition or pharmaceutical preparation described above in the preparation of a vasodilating drug or a blood pressure lowering drug.

[0090] In an eighth aspect, a method for dilating blood vessels, comprising administering the compound of the first aspect and / or the pharmaceutical composition of the second aspect to a subject in need thereof.

[0091] The present application has the following beneficial effects:

[0092] Experiments show that, at a concentration of 10 μM, the inhibition rates of WRZ-1, WRZ-2, WRZ-3, WRZ-4, WRZ-5, WRZ-6, WRZ-7, WRZ-8, WRZ-9, WRZ-14, WRZ-15, WRZ-21, WRZ-22, WRZ-24, WRZ-28, WRZ-29 provided by the present application on ROCK2 enzyme are all more than 50%, indicating that the above compounds have good ROCK2 inhibitory activity; wherein the inhibition rates of WRZ-1, WRZ-2, WRZ-3, WRZ-4, WRZ-6, WRZ-7, WRZ-8, WRZ-9, WRZ-15, WRZ-21, WRZ-22 on ROCK2 are more than 90%, which is comparable to the positive drug Fasudil; the inhibition rates of WRZ-1, WRZ-4, WRZ-6, WRZ-7 on ROCK2 are more than the inhibition rate of the positive drug Fasudil on ROCK2, showing stronger ROCK2 inhibitory activity.

[0093] Through in vivo vasodilatory activity experiments, it is shown that WRZ-1, WRZ-13, WRZ-6 provided by the present application can dilate the blood vessels induced by Phe or Phe-induced vasoconstriction.

[0094] Through activity detection on a mouse hypertension model, it is shown that WRZ-6 provided by the present application has a good effect on treating hypertension. BRIEF DESCRIPTION OF DRAWINGS

[0095] The accompanying drawings, which form a part of the present application, are included to provide a further understanding of the application, and are incorporated herein for explanation by reference. The present application will become more fully understood from the detailed description and accompanying drawings given below.

[0096] Figure 1 In the example of the present application, the compound WRZ-1 is exposed to 1x10 -6The results of vasodilatory activity induced by PE-induced vasodilation with α-adrenergic receptor agonist phenylephrine (Phe) or 60 mM KCl are shown in Figure A, B, C, and D.

[0097] Figure 2 In this embodiment of the invention, compound WRZ-13 was exposed to 1×10⁻⁶ ppm in the aortic segment of mice. -6 The results of vasodilatory activity induced by PE-induced vasodilation with α-adrenergic receptor agonist phenylephrine (Phe) or 60 mM KCl are shown in Figure A, B, C, and D.

[0098] Figure 3 In this embodiment of the invention, compound WRZ-6 was exposed to 1×10⁻⁶ ppm in the aortic segment of mice. -6 The results of vasodilatory activity induced by PE-induced vasodilation with α-adrenergic receptor agonist phenylephrine (Phe) or 60 mM KCl are shown in Figure A, B, C, and D.

[0099] Figure 4 The graph shows the results of the activity detection of compound WRZ-6 in an in vivo mouse hypertension model in the embodiments of the present invention. A is the systolic blood pressure (SBP) and B is the diastolic blood pressure (DBP).

[0100] Figure 5 The diagram shows the interaction pattern of compound WRZ-6 with ROCK2 in the embodiments of the present invention. A is the interaction pattern of WRZ-6 with key amino acids of target ROCK2, and B is the pattern of WRZ-6 extending into the active binding cavity of target ROCK2. Detailed Implementation

[0101] To enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0102] Example 1 Preparation of compounds

[0103] Compound I can be prepared via the following reaction route:

[0104]

[0105] Wherein, the reagents used are: (a) AIBN, NBS, CCl4, 78 °C, 8 h, reflux; (b) SOCl2, 80 °C, 2 h, reflux; (c) THF / DCM, Et3N, 0 °C→rt; 4-6 h;

[0106] General method for synthesis of intermediates 2a-2h

[0107] Dissolve the starting material la-lh in carbon tetrachloride, if part of the starting material cannot be completely dissolved in carbon tetrachloride, it can be completely dissolved after ultrasonic treatment. Then add 1.2 eq of N-bromosuccinimide (NBS) and 0.05 eq of azobisisobutyronitrile (AIBN), reflux at 78 °C for 8 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 1:1), after the reaction is complete, first filter to remove solid insoluble matter, retain the filtrate, concentrate, and finally silica gel column chromatography, determine the eluent polarity according to the running plate, which is usually petroleum ether: ethyl acetate 20:1 to 10:1 or so, and the intermediate 2a-2h is obtained by column chromatography.

[0108] (1) Synthesis of intermediate: 2-bromo-2-(3-methoxyphenyl)acetyl chloride (code 2a)

[0109]

[0110] Dissolve 0.83 g (5 mmol) of la and 1.1 g (6 mmol) of NBS in 20 mL of carbon tetrachloride, if the solubility of the starting material is poor, increase the reaction temperature, after the starting material is completely dissolved, add 41 mg (0.25 mmol) of AIBN, reflux at 78 °C for 8 hours or more, monitor the reaction by TLC, after the reaction is complete, filter out the insoluble matter, collect the filtrate, concentrate and then silica gel column chromatography, the eluent polarity is petroleum ether: ethyl acetate from 20:1 to 10:1, the product point is below the starting material point, but the product comes out first, 630 mg of light red oil is obtained, the yield is 51.6%.

[0111] (2) Synthesis of intermediate: 2-bromo-2-(3-fluorophenyl)acetyl chloride (code 2b)

[0112]

[0113] Dissolve 1.54 g (10 mmol) of 1b and 2.14 g (12 mmol) of NBS in 25 mL of carbon tetrachloride, if the solubility of the raw material is poor, increase the reaction temperature, after the raw material is completely dissolved, add 160 mg (1 mmol) of AIBN, reflux at 78°C for more than 8 hours, monitor the reaction by TLC, after the reaction is complete, filter out the insoluble material, retain the filtrate, concentrate the filtrate and then perform silica gel column chromatography, eluent: polar petroleum ether: ethyl acetate 10:1, obtain 2.06 g of yellow oil, yield 88%.

[0114] (3) Synthesis of intermediate: 2-bromo-2-(m-tolyl)acetyl chloride (Code 2c)

[0115]

[0116] Dissolve 600 mg (4 mmol) of 1c and 854 mg (4.8 mmol) of NBS in 20 mL of carbon tetrachloride, if the solubility of the raw material is poor, first ultrasonic, if it still cannot be completely dissolved, increase the reaction temperature, after the raw material is completely dissolved, add 66 mg (0.4 mmol) of AIBN, reflux at 78°C for more than 8 hours, monitor the reaction by TLC, the raw material is not completely reacted, directly post-treatment, filter out the insoluble material, retain the filtrate, perform silica gel column chromatography, eluent: polar petroleum ether: ethyl acetate from 20:1 to 10:1, the product point is below the raw material point, but the product comes out first, obtain 410 mg of light red oil, yield 45%.

[0117] (4) Synthesis of intermediate: 2-bromo-2-(3,4-dichlorophenyl)acetyl chloride (Code 2d)

[0118]

[0119] Dissolve 1 g (4.88 mmol) of 1d and 1.04 g (5.85 mmol) of NBS in 30 mL of carbon tetrachloride, after the raw material is completely dissolved, add 81 mg (0.49 mmol) of AIBN, reflux at 78°C for more than 8 hours, monitor the reaction by TLC, after the reaction is complete, filter out the insoluble material, obtain the filtrate, perform silica gel column chromatography, eluent: polar petroleum ether: ethyl acetate 10:1, the product point is below the raw material point, obtain 1.5 g of yellow oil, yield 65%. Directly proceed to the next step.

[0120] (5) Synthesis of intermediate: 2-bromo-2-(3-(trifluoromethyl)phenyl)acetyl chloride (Code 2e)

[0121]

[0122] Dissolve 1 g (4.9 mmol) of 1e and 1.05 g (5.88 mmol) of NBS in 25 mL of carbon tetrachloride, if the raw material solubility is poor, first ultrasonic, if still can't completely dissolve, increase the reaction temperature, after the raw material completely dissolved, add 81 mg (0.49 mmol) of AIBN, reflux at 78 degrees Celsius for more than 8 hours, TLC monitoring reaction, the raw material reaction is complete, filter out the solid insoluble, keep the filtrate, vacuum concentration after silica gel column chromatography, eluent polarity petroleum ether: ethyl acetate 10:1, the product point below the raw material point, get 1.1 g of yellow oil, yield 79%.

[0123] (6) Synthesis of intermediate 15f: 2-bromo-2-(3-chlorophenyl)acetyl chloride (Code 2f)

[0124]

[0125] Dissolve 1 g (5.86 mmol) of 1f and 1.25 g (7.03 mmol) of NBS in 25 mL of carbon tetrachloride, after the raw material completely dissolved, add 97 mg (0.59 mmol) of AIBN, reflux at 78 degrees Celsius for more than 8 hours, TLC run board monitoring reaction, this reaction is complete in a short time, the product point below the raw material point. Filter out the solid insoluble, keep the filtrate, silica gel column chromatography, eluent polarity petroleum ether: ethyl acetate 10:1, get 1.6 g of yellow oil, directly into the next step.

[0126] (7) Synthesis of intermediate: 2-bromo-2-(3,4-difluorophenyl)acetyl chloride (Code 2g)

[0127]

[0128] Dissolve 1 g (5.81 mmol) of 1g and 1.24 g (6.97 mmol) of NBS in 25 mL of carbon tetrachloride, after the raw material completely dissolved, add 96 mg (0.58 mmol) of AIBN, reflux at 78 degrees Celsius for more than 8 hours, TLC monitoring reaction, the raw material point has tailing phenomenon on the plate, the product point below the raw material point, after the reaction is complete, the tailing disappears. Filter out the solid insoluble, keep the filtrate, silica gel column chromatography, eluent polarity petroleum ether: ethyl acetate 10:1, get 1.2 g of yellow oil, directly into the next step.

[0129] (8) Synthesis of intermediate: 2-bromo-2-(3,4-difluorophenyl)acetyl chloride (Code 2h)

[0130]

[0131] Dissolve 1.5 g (7.65 mmol) of 1h and 1.63 g (9.2 mmol) of NBS in 25 mL of carbon tetrachloride, if the raw material solubility is poor, increase the reaction temperature, after the raw material is completely dissolved, add 125 mg (0.76 mmol) of AIBN, reflux at 78 degrees Celsius for more than 8 hours, monitor the reaction by TLC, this reaction is not complete, which affects the yield. Directly post-processing, filter out the insoluble, retain the filtrate, silica gel column chromatography, eluent polarity petroleum ether: ethyl acetate from 20: 1 to 10: 1, the product point is below the raw material point, get yellow oil, yield 40%

[0132] General method for synthesis of intermediates 3a-3h

[0133] At 0°C (ice bath), 10 eq of thionyl chloride is added dropwise to the raw material 2a-2h, after the addition is complete, react at room temperature for 5 to 10 minutes, then reflux for about 2 hours, concentrate under vacuum, spin out the thionyl chloride, and obtain the intermediate 3a-3h as a yellow oil. Another method is to dissolve 14a-14h in dichloromethane, then add 7-8 equivalents of oxalyl chloride and 3-5 drops of DMF, reflux at 38°C for two hours, concentrate the solution after the reaction is complete, and directly proceed to the next step.

[0134] (9) Synthesis of intermediate: 2-bromo-2-(3-methoxyphenyl)acetyl chloride (code 3a)

[0135]

[0136] Under ice bath, 1.9 g (15.9 mmol) of thionyl chloride is added dropwise to 390 mg (1.59 mmol) of intermediate 2a, after the addition is complete, react at room temperature for 10 minutes, then reflux in a constant temperature oil bath for 2 hours, after the reaction is complete, spin off the thionyl chloride, and obtain a red oil with a yield of 85%, which is directly used in the next step.

[0137] (10) Synthesis of intermediate: 2-bromo-2-(3-fluorophenyl)acetyl chloride (code 3b)

[0138]

[0139] Under ice bath, 10.5 g (88.8 mmol) of thionyl chloride is added dropwise to 2.06 mg (8.88 mmol) of intermediate 2b, after the addition is complete, react at room temperature for 10 minutes, then reflux in a constant temperature oil bath for 2 hours, after the reaction is complete, spin off the thionyl chloride, and obtain a red oil with a yield of 90%, which is directly used in the next step.

[0140] (11) Synthesis of intermediate: 2-bromo-2-(m-tolyl)acetyl chloride (code 3c)

[0141]

[0142] Dissolve 720 mg (3.16 mmol) of intermediate 2c in 20 mL of dichloromethane, then add 3.2 g (25.7 mmol) of oxalyl chloride, reflux at 38°C for 2 hours, after the reaction is completed, concentrate the reaction solution, directly cast to the next step.

[0143] (12) Synthesis of intermediate: 2-bromo-2-(3,4-dichlorophenyl)acetyl chloride (Code 3d)

[0144]

[0145] Dissolve 1.5 g (5.3 mmol) of intermediate 2d in 20 mL of dichloromethane, then add 5.4 g (42.4 mmol) of oxalyl chloride, reflux at 38°C for 2 hours, after the reaction is completed, concentrate the reaction solution to obtain 1.17 g of red oil, directly cast to the next step.

[0146] (13) Synthesis of intermediate: 2-bromo-2-(3-(trifluoromethyl)phenyl)acetyl chloride (Code 3e)

[0147]

[0148] Dissolve 967 mg (3.43 mmol) of intermediate 2e in 20 mL of dichloromethane, then add 3.48 g (27.44 mmol) of oxalyl chloride, reflux at 38°C for 2 hours, after the reaction is completed, concentrate the reaction solution, directly cast to the next step.

[0149] (14) Synthesis of intermediate 15f: 2-bromo-2-(3-chlorophenyl)acetyl chloride (Code 3f)

[0150]

[0151] Dissolve 1.01 g (4.6 mmol) of intermediate 2f in 20 mL of dichloromethane, then add 4.7 g (36.8 mmol) of oxalyl chloride, reflux at 38°C for 2 hours, after the reaction is completed, concentrate the reaction solution, directly cast to the next step.

[0152] (15) Synthesis of intermediate: 2-bromo-2-(3,4-difluorophenyl)acetyl chloride (Code 3g)

[0153]

[0154] Dissolve 1.45 g (5.8 mmol) of intermediate 2g in 25 mL of dichloromethane, then add 5.9 g (46.4 mmol) of oxalyl chloride, reflux at 38°C for 2 hours, after the reaction is completed, concentrate the reaction solution to obtain 1.36 g of red oil, directly cast to the next step.

[0155] (16) Synthesis of intermediate: 2-bromo-2-(3,4-difluorophenyl)acetyl chloride (Code 3h)

[0156]

[0157] Dissolve 1.06 g (3.87 mmol) of intermediate 2h in 20 mL of dichloromethane, add 4 g (30.96 mmol) of oxalyl chloride, drop 5 drops of DMF, reflux at 38 °C for 2 hours, concentrate the reaction solution to get red oil, yield 92%, directly to next step.

[0158] General method for synthesis of intermediates 4a-4h

[0159] At 0 °C (ice bath), slowly drop 10 eq of thionyl chloride into starting material 1a-1h, after drop completion, react at room temperature for 10 minutes, then reflux for about 2 hours, concentrate under vacuum, spin out thionyl chloride, get yellow oil of intermediate 4a-4h. Another method is to dissolve 1a-1h in dichloromethane, add 7-8 eq of oxalyl chloride and 3-5 drops of DMF, reflux at 38 °C for 2 hours, concentrate the solution after reaction is complete, directly to next step.

[0160] (17) Synthesis of intermediate: 2-(3-methoxyphenyl)acetyl chloride (Code 4a)

[0161]

[0162] Dissolve 1 g (6 mmol) of starting material 1a in 45 mL of dichloromethane, add 5.71 g (45 mmol) of oxalyl chloride and 5 drops of DMF, reflux at 38 °C for 2 hours or more, monitor the reaction by TLC, concentrate the reaction solution after reaction is complete, get yellow oil, yield 90%, directly to next step.

[0163] (18) Synthesis of intermediate: 2-(3-methoxyphenyl)acetyl chloride (Code 4b)

[0164]

[0165] Dissolve 770 mg (5 mmol) of starting material 1b in 25 mL of dichloromethane, add 4.75 g (37.5 mmol) of oxalyl chloride and 5 drops of DMF, reflux at 38 °C for 2 hours or more, monitor the reaction by TLC, concentrate the reaction solution after reaction is complete, get yellow oil, yield 95%, directly to next step.

[0166] (19) Synthesis of intermediate: 2-(m-tolyl)acetyl chloride (Code 4c)

[0167]

[0168] Dissolve 600 mg (4 mmol) of starting material 1c in 20 mL of dichloromethane, add 3.8 g (30 mmol) of oxalyl chloride and 5 drops of DMF, reflux at 38 °C for more than 2 hours, monitor the reaction by TLC, after the reaction is complete, concentrate the reaction solution to obtain yellow oil, the yield is 85%, directly put into the next step.

[0169] (20) Synthesis of intermediate: 2-(3,4-dichlorophenyl)acetyl chloride (Code 4d)

[0170]

[0171] Dissolve 1.02 g (5 mmol) of starting material 1d in 30 mL of dichloromethane, add 4.75 g (37.5 mmol) of oxalyl chloride and 5 drops of DMF, reflux at 38 °C for more than 2 hours, monitor the reaction by TLC, after the reaction is complete, concentrate the reaction solution to obtain yellow oil, the yield is 88%, directly put into the next step.

[0172] (21) Synthesis of intermediate: 2-(3-(trifluoromethyl)phenyl)acetyl chloride (Code 4e)

[0173]

[0174] Dissolve 816 mg (4 mmol) of starting material 1e in 25 mL of dichloromethane, add 3.81 g (30 mmol) of oxalyl chloride and 4 drops of DMF, reflux at 38 °C for more than 2 hours, monitor the reaction by TLC, after the reaction is complete, concentrate the reaction solution to obtain yellow oil, the yield is 90%, directly put into the next step.

[0175] (22) Synthesis of intermediate: 2-(3-(trifluoromethyl)phenyl)acetyl chloride (Code 4f)

[0176]

[0177] Dissolve 850 mg (5 mmol) of starting material 1e in 30 mL of dichloromethane, add 4.75 g (37.5 mmol) of oxalyl chloride and 4 drops of DMF, reflux at 38 °C for more than 2 hours, monitor the reaction by TLC, after the reaction is complete, concentrate the reaction solution to obtain yellow oil, the yield is 92%, directly put into the next step.

[0178] (23) Synthesis of intermediate: 2-(3,4-difluorophenyl)acetyl chloride (Code 4g)

[0179]

[0180] Dissolve 688 mg (4 mmol) of the starting material 1g in 20 mL of dichloromethane, add 3.81 g (30 mmol) of oxalyl chloride and 4 drops of DMF, reflux at 38°C for more than 2 hours, monitor the reaction by TLC, after the reaction is completed, concentrate the reaction solution to obtain yellow oil, the yield is 96%, directly proceed to the next step.

[0181] (24) Synthesis of intermediate: 2-(3,4-dimethoxyphenyl)acetyl chloride (Code 4h)

[0182]

[0183] Dissolve 1 g (5.09 mmol) of the starting material 1h in 20 mL of dichloromethane, add 4.85 g (38.22 mmol) of oxalyl chloride and 5 drops of DMF, reflux at 38°C for more than 2 hours, monitor the reaction by TLC, after the reaction is completed, concentrate the reaction solution to obtain yellow oil, the yield is 94%, directly proceed to the next step.

[0184] Synthesis method of WRZ-1 to WRZ-12:

[0185] Dissolve 5-aminoindazole in dichloromethane or tetrahydrofuran and add 2.4 eq of triethylamine, then dissolve 1.2 eq of intermediate 3a-3h or intermediate 4a-4h in an appropriate amount of dichloromethane or tetrahydrofuran, add the solution of intermediate 3a-3h or 4a-4h dropwise to the primary amine solution, react for more than 4 hours, monitor the reaction by TLC (dichloromethane:methanol = 20:1), after the reaction is completed, extract with dichloromethane three times (appropriate amount of DCM), combine the organic phases, remove water from the organic phase with anhydrous magnesium sulfate, and purify by silica gel column chromatography to obtain pure WRZ-1 to WRZ-12.

[0186] (25) Synthesis of target compound: N-(1H-indazol-5-yl)-2-(3-methoxyphenyl)acetamide (Code WRZ-1)

[0187]

[0188] 280 mg (2.1 mmol) of 5-aminoindazole, 463 mg (2.52 mmol) of intermediate 4a, and 510 mg (5.04 mmol) of triethylamine. According to the general synthesis method described above, 300 mg of white solid was obtained, the yield was 51%. 1H NMR (800 MHz, DMSO-d6): δ 12.97 (s, NH), 10.14 (s, NH), 8.12 (s, ArH), 8.01 (s, ArH), 7.48 (d, J = 8.9 Hz, ArH), 7.43 (d, J = 6.9 Hz, 1H), 7.24 (t, J = 7.9 Hz, 1H), 6.96 - 6.92 (m, 2H), 6.82 (dd, J = 8.2, 2.8 Hz, 1H), 3.75 (s, CH3), 3.62 (s, CH2); 13 C NMR (201 MHz, DMSO-d6): δ 169.19, 159.68, 138.11, 137.33, 133.84, 132.67, 129.78, 123.16, 121.76, 120.74, 115.34, 112.33, 110.62, 110.10, 55.44, 43.84; ESI-MS (m / z): calcd for C 16 H 15 N3O2[M+H] + 282.12, found 282.23.

[0189] (26) Synthesis of the target compound: 2-(3-fluorophenyl)-N-(1H-indazol-5-yl)acetamide (Code: WRZ-2)

[0190]

[0191] Dissolve 380 mg (2.91 mmol) of 5-aminoindazole in 20 mL of tetrahydrofuran, and add 587 mg (5.82 mmol) of triethylamine to obtain an indazole solution. Dissolve 560 mg (3.5 mmol) of intermediate 4b in 10 mL of tetrahydrofuran, and drop the THF solution of intermediate 4b into the indazole solution, and react at room temperature for 8 hours, monitoring once every two hours by TLC. After the reaction is completed, first filter out the insoluble matter, concentrate the filtrate, then add 20 mL of dichloromethane, and the product is insoluble and precipitates as a solid. Then purify by silica gel column chromatography to obtain the pure target compound as a white solid, 460 mg, in a yield of 49.5%. 1 H NMR (800 MHz, DMSO-d6): δ 12.99 (s, NH), 10.19 (s, NH), 8.13 (s, ArH), 8.02 (s, arH), 7.49 (d, J = 8.9 Hz, 1H), 7.43 (d, J = 10.8 Hz, 1H), 7.38 (q, J = 7.5 Hz, 1H), 7.20 (d, J = 7.1 Hz, 2H), 7.09 (t, J = 8.0 Hz, 1H), 3.70 (s, CH2); 13C NMR (201 MHz, DMSO-d6): δ 168.78, 163.14, 161.93, 139.35, 137.37, 133.86, 132.56, 130.57, 125.76, 123.16, 120.75, 116.43, 113.84, 110.65, 110.19, 43.26; ESI-MS (m / z): calcd for C 15 H 12 FN3O[M+H] + 270.10, found 270.25.

[0192] (27) The synthesis of the target compound: N-(1H-indazol-5-yl)-2-(m-tolyl)acetamide (Code: WRZ-3)

[0193]

[0194] Dissolve 310 mg (2.33 mmol) of 5-aminoindazole in 30 mL of tetrahydrofuran, and add 566 mg (5.6 mmol) of triethylamine to obtain an indazole solution. Dissolve 470 mg (2.8 mmol) of intermediate 4c in 10 mL of tetrahydrofuran, and drop the THF solution of intermediate 4c into the indazole solution, and react at room temperature for about 8 hours, monitoring once every two hours by TLC. After the reaction is completed, first filter out the insoluble matter, concentrate the filtrate, and then perform silica gel column chromatography to obtain the pure target compound as a white solid 360 mg, with a yield of 74%. 1 H NMR (800 MHz, DMSO-d6): δ 12.97 (s, NH), 10.14 (s, NH), 8.13 (s, ArH), 8.01 (s, ArH), 7.50-7.42 (m, 2H), 7.24-7.14 (m, 3H), 7.06 (d, J = 7.6 Hz, 1H), 3.61 (s, CH2), 2.30 (s, CH3); 13 C NMR (201 MHz, DMSO-d6): δ 169.38, 137.79, 137.32, 136.56, 133.84, 132.71, 130.17, 128.67, 127.59, 126.60, 123.17, 120.73, 110.61, 110.07, 43.76, 21.47; ESI-MS (m / z): calcd for C 16 H 15 N3O[M+H] + 266.12, found 266.77.

[0195] (28) Synthesis of the target compound: N-(1H-indazol-5-yl)-2-(3- (trifluoromethyl)phenyl)acetamide (Code WRZ-4)

[0196]

[0197] 240 mg (1.8 mmol) of 5-aminoindazole, 480 mg (2.16 mmol) of intermediate 4e, 437 mg (4.32 mmol) of triethylamine. White solid 310 mg was obtained according to the general synthesis method, yield 54%. 1 H NMR (800 MHz, DMSO-d6): δ 12.99 (s, NH), 10.25 (s, NH), 8.13 (s, ArH), 8.02 (s, ArH), 7.74 (s, ArH), 7.68-7.57 (m, 3H), 7.51-7.43 (m, 2H), 3.81 (s, CH2); 13 C NMR (201 MHz, DMSO-d6): δ 168.72, 138.00, 137.39, 133.87, 132.52, 129.77, 129.51, 129.36, 126.24, 125.45, 124.09, 123.76, 123.16, 120.73, 110.68, 110.21, 43.09; ESI-MS (m / z): calcd. for C 16 H 12 F3N3O[M+H] + 320.09, found 321.1

[0198] (29) Synthesis of the target compound: 2-(3,4-dimethoxyphenyl)-N-(1H- indazol-5-yl)acetamide (Code WRZ-5)

[0199]

[0200] 450 mg (3.82 mmol) of 5-aminoindazole was dissolved in 35 mL of tetrahydrofuran, and 1.08 g (10.68 mmol) of triethylamine was added to obtain an indazole solution. 0.98 g (4.58 mmol) of intermediate 4h was dissolved in 10 mL of tetrahydrofuran, and the tetrahydrofuran solution of intermediate 4h was added dropwise to the indazole solution, and the reaction was carried out at room temperature for 8 hours, and TLC was monitored every two hours. After the reaction was completed, the insoluble matter was first filtered out, the filtrate was concentrated, then 20 mL of ethyl acetate was added, the product was insoluble, and a solid was precipitated, which was confirmed as a crude product by liquid chromatography-mass spectrometry, and 750 mg of white solid was obtained by silica gel column chromatography, with a yield of 63%. 1HNMR (800 MHz, DMSO-d6): δ 12.97 (s, 1H), 10.09 (s, 1H), 8.12 (s, 1H), 8.01 (s, 1H), 7.48 (d, J = 8.9 Hz, 1H), 7.43 (d, J = 10.3 Hz, 1H), 6.98 (s, 1H), 6.92 - 6.85 (m, 2H), 3.74 (d, J = 23.4 Hz, 6H), 3.57 (s, 2H); 13 C NMR (201 MHz, DMSO-d6): δ 169.60, 148.99, 148.05, 137.32, 133.82, 132.74, 129.03, 123.16, 121.50, 120.77, 113.47, 112.30, 110.60, 110.09, 56.00, 55.91, 43.37; ESI-MS (m / z): calcd for C 17 H 17 N3O3[M+H] + 312.13, found 312.33.

[0201] (30) Synthesis of the target compound: 2-bromo-N-(1H-indazol-5-yl)-2-(3- methoxyphenyl)acetamide (Code: WRZ-6)

[0202]

[0203] Dissolve 150 mg (1.12 mmol) of 5-aminoindazole in 20 mL of tetrahydrofuran, and add 260 mg (2.56 mmol) of triethylamine to obtain an indazole solution. Dissolve 335 mg (1.28 mmol) of intermediate 3a in 10 mL of tetrahydrofuran, and dropwise add the THF solution of intermediate 3a to the indazole solution, and react at room temperature for 8 hours, monitoring once every two hours by TLC. After the reaction is completed, first filter off the insoluble matter, concentrate the filtrate, and purify by silica gel column chromatography to obtain 143 mg of the target compound. 1 H NMR (800 MHz, DMSO-d6): δ 12.97 (s, 1H), 10.09 (s, 1H), 8.12 (s, 1H), 8.01 (s, 1H), 7.48 (d, J = 8.9 Hz, 1H), 7.43 (d, J = 10.3 Hz, 1H), 6.98 (s, 1H), 6.92 - 6.85 (m, 2H), 3.74 (d, J = 23.4 Hz, 6H), 3.57 (s, 2H); 13C NMR (201 MHz, DMSO-d6): δ 169.67, 159.69, 140.80, 137.89, 137.82, 133.76, 129.81, 121.80, 121.11, 119.59, 115.40, 114.50, 112.38, 99.30, 55.44, 43.95; ESI-MS (m / z): calcd for C 16 H 14 BrN3O2[M+H] + 361.02, found 361.39.

[0204] (31) Synthesis of the target compound: 2-bromo-2-(3-fluorophenyl)-N-(1H-indazol-5- yl)acetamide (Code: WRZ-7)

[0205]

[0206] Dissolve 500 mg (4.4 mmol) of 5-aminoindazole in 30 mL of tetrahydrofuran, and add 1.1 g (10.64 mmol) of triethylamine to obtain an indazole solution. Dissolve 1.33 g (3.5 mmol) of intermediate 3b in 10 mL of tetrahydrofuran, and drop the tetrahydrofuran solution of intermediate 3b into the indazole solution, and react at room temperature for 8 hours, monitoring once every two hours by TLC. After the reaction is completed, first filter out the insoluble matter, concentrate the filtrate, then add 20 mL of ethyl acetate, and precipitate 850 mg of a solid, which is confirmed by liquid chromatography-mass spectrometry to be the crude product. Recrystallize the crude product from ethyl acetate and petroleum ether to obtain 600 mg of a solid product, with a yield of 40%. 1 H NMR (800 MHz, DMSO-d6): δ 12.99 (s, NH), 10.19 (s, NH), 8.13 (s, ArH), 8.02 (s, ArH), 7.49 (d, J = 8.9 Hz, ArH), 7.44 (d, J = 8.9 Hz, 1H), 7.38 (q, J = 7.5 Hz, 1H), 7.20 (d, J = 7.3 Hz, 2H), 7.09 (t, J = 8.6 Hz, 1H); 13 C NMR (201 MHz, DMSO-d6): δ 168.74, 163.19, 161.93, 139.39, 139.35, 137.37, 133.86, 132.56, 130.61, 130.57, 125.76, 125.75, 123.18, 120.75, 116.43, 116.33, 113.84, 113.79, 110.65, 110.19, 43.26; ESI-MS (m / z): calcd for C 15 H11 BrN3O[M+H] + 349.01, found 349.21.

[0207] (32) Synthesis of the target compound: 2-bromo-N-(lH-indazol-5-yl)-2-(m-tolyl)acetamide (Code: WRZ-8)

[0208]

[0209] Dissolve 190 mg (1.43 mmol) of 5-aminoindazole in 20 mL of tetrahydrofuran, and add 320 mg (3.16 mmol) of triethylamine to obtain an indazole solution. Dissolve 1.33 g (3.5 mmol) of intermediate 3c in 10 mL of tetrahydrofuran, and drop the tetrahydrofuran solution of intermediate 3c into the indazole solution, and react at room temperature for 8 hours, monitoring once every two hours by TLC. After the reaction is completed, first filter out the insoluble matter, concentrate the filtrate, extract with dichloromethane three times, combine the organic phases, remove water from the organic phase with anhydrous magnesium sulfate, concentrate, add EA, and precipitate a solid. After filtering the filtrate, column chromatography on silica gel gives 200 mg of the target compound, with a yield of 41%. 1 H NMR (800 MHz, DMSO-d6): δ 9.92 (s, NH), 8.17 (s, NH), 8.02 (s, 1H), 7.58 (d, J = 9.0 Hz, 1H), 7.52 (dd, J = 9.2, 3.7 Hz, 2H), 7.47 (d, J = 9.0 Hz, 2H), 7.24 (t, J = 7.6 Hz, 2H), 7.10 (d, J = 7.6 Hz, 1H), 2.31 (s, CH3); 13 C NMR (201 MHz, DMSO-d6): δ 169.38, 137.79, 137.32, 136.56, 133.84, 132.71, 130.17, 128.67, 127.59, 126.60, 123.17, 120.73, 110.61, 110.07, 43.76, 21.47; ESI-MS (m / z): calcd. for C 16 H 14 BrN3O[M+H] + 345.03, found 345.29.

[0210] (33) Synthesis of the target compound: 2-bromo-2-(3,4-dichlorophenyl)-N-(lH-indazol-5-yl)acetamide (Code: WRZ-9)

[0211]

[0212] Dissolve 250 mg (1.88 mmol) of 5-aminoindazole in 30 mL of tetrahydrofuran, and add 460 mg (4.5 mmol) of triethylamine to obtain an indazole solution. Dissolve 676 mg (2.26 mmol) of intermediate 3d in 10 mL of tetrahydrofuran, and drop the THF solution of intermediate 3d into the indazole solution, and react at room temperature for 8 hours with magnetic stirring, and monitor the reaction by TLC. After the raw material is completely reacted, first suction-filter out the solid insoluble matter, concentrate the filtrate, and subject to silica gel column chromatography (dichloromethane:methanol = 200:1) to obtain 320 mg of the target compound, with a yield of 43%. 1 H NMR (800 MHz, DMSO-d6): δ 10.04 (s, NH), 8.16 (s, NH), 8.03 (s, ArH), 7.80 (s, ArH), 7.65 (d, J = 8.4 Hz, ArH), 7.56 (dd, J = 22.0, 8.7 Hz, ArH), 7.49 (d, J = 9.0 Hz, CH); 13 C NMR (201 MHz, DMSO-d6): δ 168.72, 138.00, 137.39, 133.87, 132.52, 129.77, 129.51, 129.36, 126.24, 125.45, 124.09, 123.76, 123.16, 120.73, 110.68, 110.21, 43.09; ESI-MS (m / z): calcd. for C 15 H 10 BrCl2N3O [M+H] + 398.93, found 399.21.

[0213] (34) Synthesis of the target compound: 2-bromo-N-(1H-indazol-5-yl)-2-(3- (trifluoromethyl)phenyl)acetamide (Code: WRZ-10)

[0214]

[0215] Dissolve 300 mg (2.28 mmol) of 5-aminoindazole in 30 mL of tetrahydrofuran, and add 550 mg (5.48 mmol) of triethylamine to obtain an indazole solution. Dissolve 807 mg (2.74 mmol) of intermediate 3e in 10 mL of tetrahydrofuran, and drop the THF solution of intermediate 3e into the indazole solution, and react at room temperature for 8 hours with magnetic stirring, and monitor the reaction by TLC. After the reaction is complete, first suction-filter out the solid insoluble matter, retain the filtrate, concentrate, and subject to silica gel column chromatography (dichloromethane:methanol = 200:1) to obtain 290 mg of the target compound, with a yield of 40%. 1H NMR (800 MHz, DMSO-d6): δ 10.08 (s, NH), 8.17 (s, NH), 8.06 (s, ArH), 8.03 (s, ArH), 7.93 (s, ArH), 7.86 (d, J = 7.9 Hz, 1H), 7.68 (d, J = 7.4 Hz, 1H), 7.62 (t, J = 7.8 Hz, 1H), 7.58 (d, J = 7.0 Hz, 1H), 7.49 (d, J = 8.9 Hz, 1H), 5.30 (s, CH); 13 C NMR (201 MHz, DMSO-d6): δ 169.38, 137.79, 137.32, 136.56, 133.84, 132.71, 130.17, 128.67, 127.59, 126.60, 123.17, 120.73, 110.61, 110.07, 43.76, 21.47; ESI-MS (m / z): calcd for C 16 H 11 BrF3N3O [M+H] + 399.01, found 399.73.

[0216] (35) Synthesis of the target compound 2-bromo-2-(3-chlorophenyl)-N-(lH-indazol-5- yl)acetamide (Code WRZ-11)

[0217]

[0218] Dissolve 370 mg (2.8 mmol) of 5-aminoindazole in 30 mL of tetrahydrofuran, and add 793 mg (7.84 mmol) of triethylamine to obtain an indazole solution. Dissolve 897 mg (3.36 mmol) of intermediate 3f in 10 mL of tetrahydrofuran, and dropwise add the THF solution of intermediate 3f to the indazole solution, and react at room temperature for 8 hours with stirring, and monitor the reaction by TLC. After the reaction is completed, first filter out the solid insoluble matter, concentrate the filtrate, and perform silica gel column chromatography (dichloromethane:methanol = 200:1) to obtain 310 mg of the target compound, with a yield of 31%. 1 H NMR (800 MHz, DMSO-d6): δ 10.08 (s, NH), 8.17 (s, NH), 8.06 (s, ArH), 8.03 (s, ArH), 7.93 (s, ArH), 7.86 (d, J = 7.9 Hz, 1H), 7.68 (d, J = 7.4 Hz, 1H), 7.62 (t, J = 7.8 Hz, 1H), 7.58 (d, J = 7.0 Hz, 1H), 7.49 (d, J = 8.9 Hz, 1H), 5.30 (s, CH); 13C NMR (201 MHz, DMSO-d6): d 168.78, 163.14, 161.93, 139.35, 137.37, 133.86, 132.56, 130.57, 125.76, 123.16, 120.75, 116.43, 113.84, 111.75, 110.59, 44.36; ESI-MS (m / z): calcd for C 15 H 11 BrClN3O[M+H] + 364.97, found 365.21.

[0219] (36) Synthesis of the target compound: 2-bromo-2-(3,4-difluorophenyl)-N-(1H-indazol-5- yl)acetamide (Code: WRZ-12)

[0220]

[0221] Dissolve 260 mg (2.5 mmol) of 5-aminoindazole in 30 mL of tetrahydrofuran, and add 753 mg (7.46 mmol) of triethylamine to obtain an indazole solution. Dissolve 1 g (3.73 mmol) of intermediate 3g in 10 mL of tetrahydrofuran, and drop the THF solution of intermediate 3g into the indazole solution, and react at room temperature for 8 hours with stirring, and monitor the reaction by TLC. After the reaction is completed, first filter out the solid undissolved substance, concentrate the filtrate, and perform silica gel column chromatography (dichloromethane:methanol = 200:1) to obtain 240 mg of the target compound, with a yield of 27%. 1 H NMR (800 MHz, DMSO-d6): d 10.02 (s, NH), 8.16 (s, NH), 8.03 (s, ArH), 7.60-7.56 (m, ArH), 7.53 (d, J = 8.9 Hz, 1H), 7.49 (d, J = 9.0 Hz, 1H), 7.44 (dt, J = 17.8, 9.1 Hz, 2H), 7.40 (d, J = 5.0 Hz, CH); 13 C NMR (201 MHz, DMSO-d6): d 168.78, 163.14, 161.93, 139.35, 137.37, 133.86, 132.56, 130.57, 125.76, 123.16, 120.75, 116.43, 112.78, 110.68, 109.19, 42.36; ESI-MS (m / z): calcd for C 15 H 10 BrF2N3O[M+H] + 366.99, found 367.49.

[0222] The compound of formula II can be prepared by the following reaction route:

[0223]

[0224] Wherein, the reagents used are respectively: (a) AIBN, NBS, CCl4, 78°C, 8h, reflux; (b) SOCl2, 80°C, 2h, reflux; (c) THF / DCM, Et3N, 0°C→rt; 4-6h;

[0225] Specifically, the present application gives the preparation process of some schematic compounds and effect verification data in the examples.

[0226] (37) Synthesis of 2-(3-methoxyphenyl)-N-(quinolin-6-yl)acetamide (Code: WRZ-13)

[0227]

[0228] Dissolve 200 mg (1.38 mmol) of 6-aminoquinoline in 20 mL of dichloromethane, and add 335 mg (3.31 mmol) of triethylamine to obtain a quinoline solution. Dissolve 304 mg (1.65 mmol) of intermediate 4a in 10 mL of dichloromethane, and drop the dichloromethane solution of intermediate 4a into the quinoline solution, and react overnight. TLC monitoring shows that the reaction is complete. After the reaction is complete, the product is obtained by silica gel column chromatography and recrystallization, with a yield of 241 mg, 60%. 1 H NMR (800 MHz, DMSO-d6): δ 11.26 (s, 1H), 9.13 (d, J = 5.2 Hz, 1H), 9.04 (d, J = 8.5 Hz, 1H), 8.77 (s, 1H), 8.37 (d, J = 9.2 Hz, 1H), 8.23 (d, J = 9.2 Hz, 1H), 7.98 (dd, J = 8.5, 5.0 Hz, 1H), 7.25 (t, J = 7.9 Hz, 1H), 7.03-6.96 (m, 2H), 6.83 (d, J = 11.0 Hz, 1H), 3.78 (s, 2H), 3.75 (s, 3H); 13 C NMR (201 MHz, DMSO-d6): δ 170.62, 159.69, 144.99, 143.83, 140.07, 137.49, 135.74, 129.91, 129.81, 128.04, 122.80, 122.73, 121.89, 115.51, 115.28, 112.52, 55.48, 43.76; ESI-MS (m / z): calcd. for C 18 H 16N2O2[M+H] + 293.12, found 293.25.

[0229] (38) Synthesis of target compound 2-(3-fluorophenyl)-N-(quinolin-6-yl)acetamide (Code: WRZ-14)

[0230]

[0231] To a solution of 400 mg (2.91 mmol) of 6-aminoquinoline in 20 mL of dichloromethane, 587 mg (5.8 mmol) of triethylamine was added to obtain a quinoline solution. 560 mg (3.5 mmol) of intermediate 4b was dissolved in 10 mL of dichloromethane, and the dichloromethane solution of intermediate 4b was added dropwise to the quinoline solution, and the reaction was allowed to proceed overnight. A large amount of solid was precipitated from the reaction solution, which was dissolved and TLC was run (dichloromethane:methanol = 20:1) to find two spots, the upper one was confirmed to be the product spot by LC-MS, and the lower one was the starting material spot. However, the starting material was dissolved in dichloromethane. Silica gel column chromatography was performed to obtain 310 mg of white solid, with a yield of 40%. 1 H NMR (800 MHz, DMSO-d6): δ 11.35 (s, NH), 9.12 (d, J = 5.2 Hz, ArH), 9.02 (d, J = 8.4 Hz, ArH), 8.76 (s, ArH), 8.37 (d, J = 9.1 Hz, 1H), 8.23 (d, J = 6.6 Hz, 1H), 7.97 (dd, J = 8.5, 4.9 Hz, 1H), 7.39 (q, J = 7.6 Hz, 1H), 7.26 (dd, J = 18.2, 9.0 Hz, 2H), 7.13 - 7.08 (m, 1H), 3.87 (s, CH2); 13 C NMR (201 MHz, DMSO-d6): δ 170.21, 163.11, 161.90, 144.72, 144.05, 139.91, 138.76, 136.06, 130.65, 129.86, 127.89, 125.93, 122.74, 116.62, 115.35, 113.91, 43.18; ESI-MS (m / z): calcd. for C 17 H 13 FN2O[M+H] + 281.10, found 281.54.

[0232] (39) Synthesis of target compound N-(quinolin-6-yl)-2-(m-tolyl)acetamide (Code: WRZ-15)

[0233]

[0234] To a solution of 150 mg (1.04 mmol) of 6-aminoquinoline in 20 mL of dichloromethane, 253 mg (2.5 mmol) of triethylamine was added to obtain a quinoline solution. 344 mg (1.25 mmol) of intermediate 4c was dissolved in 10 mL of dichloromethane, and the dichloromethane solution of intermediate 4c was added dropwise to the quinoline solution, and the reaction was stirred magnetically overnight, and the reaction was monitored by TLC, and after the raw material was completely reacted, the product 175 mg was obtained by silica gel column chromatography and recrystallization, with a yield of 61%. 1 H NMR (800 MHz, DMSO-d6): δ 10.50 (s, 1H), 8.78 (d, J = 4.2 Hz, 1H), 8.39 (d, J = 2.6 Hz, 1H), 8.26 (d, J = 10.1 Hz, 1H), 7.98 (d, J = 9.1 Hz, 1H), 7.82 (dd, J = 9.1, 2.5 Hz, 1H), 7.47 (dd, J = 8.3, 4.1 Hz, 1H), 7.23 (t, J = 7.6 Hz, 1H), 7.20 - 7.16 (m, 2H), 7.07 (d, J = 7.5 Hz, 1H), 3.68 (s, 2H), 2.30 (s, 3H); 13 C NMR (201 MHz, DMSO-d6): δ 170.10, 149.46, 145.13, 137.85, 137.56, 136.17, 135.95, 130.23, 129.97, 128.78, 128.72, 127.71, 126.67, 123.74, 122.24, 115.39, 43.80, 21.47; ESI-MS (m / z): calcd. for C 18 H 16 N2O[M+H] + 277.13, found 277.50.

[0235] (40) Synthesis of the target compound 2-(3,4-dichlorophenyl)-N-(quinolin-6-yl)acetamide (Code WRZ-16)

[0236]

[0237] To a solution of 200 mg (1.38 mmol) of 6-aminoquinoline in 20 mL of dichloromethane, 335 mg (3.31 mmol) of triethylamine was added to obtain a quinoline solution. 304 mg (1.65 mmol) of intermediate 4d was dissolved in 10 mL of dichloromethane, and the dichloromethane solution of intermediate 4d was added dropwise to the quinoline solution, and the reaction was allowed to proceed overnight, and the reaction was monitored by TLC. After the starting material was completely reacted, the product 241 mg was obtained by silica gel column chromatography and recrystallization, with a yield of 60%, white solid, and a yield of 65%. 1 H NMR (800 MHz, DMSO-d6): δ 10.55 (s, NH), 8.79 (d, J = 4.1 Hz, ArH), 8.38 (d, J = 2.6 Hz, ArH), 8.28 (s, ArH), 7.98 (d, J = 9.0 Hz, ArH), 7.81 (dd, J = 9.0, 2.5 Hz, 1H), 7.66 (s, 1H), 7.61 (d, J = 8.2 Hz, 1H), 7.48 (dd, J = 8.3, 4.1 Hz, 1H), 7.37 (d, J = 8.2 Hz, 1H), 3.79 (s, CH2); 13 C NMR (201 MHz, DMSO-d6): δ 169.22, 149.55, 145.18, 137.33, 135.97, 131.86, 131.21, 130.85, 130.25, 130.03, 129.82, 128.77, 123.71, 122.27, 115.54, 49.07, 42.39; ESI-MS (m / z): calcd. for C 17 H 12 Cl2N2O[M+H] + 332.02, found 332.77.

[0238] (41) Synthesis of the target compound N-(quinolin-6-yl)-2-(3-(trifluoromethyl)phenyl)acetamide (Code WRZ-17)

[0239]

[0240] To a solution of 270 mg (1.87 mmol) of 6-aminoquinoline in 20 mL of dichloromethane, 455 mg (4.5 mmol) of triethylamine was added to obtain a quinoline solution. 497 mg (2.24 mmol) of intermediate 4e was dissolved in 10 mL of dichloromethane, and the dichloromethane solution of intermediate 4e was added dropwise to the quinoline solution, and the reaction was allowed to proceed overnight, and the reaction was monitored by TLC. After the starting material was completely reacted, the product 350 mg was obtained by silica gel column chromatography and recrystallization, with a yield of 58%. 1H NMR (800 MHz, DMSO-d6): δ 10.59 (s, NH), 8.79 (d, J = 2.5 Hz, ArH), 8.39 (d, J = 2.6 Hz, ArH), 8.27 (d, J = 6.6 Hz, ArH), 7.99 (d, J = 9.1 Hz, 1H), 7.82 (dd, J = 9.1, 2.5 Hz, 1H), 7.76 (s, 1H), 7.69 (d, J = 7.6 Hz, 1H), 7.65 (d, J = 8.0 Hz, 1H), 7.60 (t, J = 7.7 Hz, 1H), 7.48 (dd, J = 8.3, 4.1 Hz, 1H), 3.89 (s, CH2); 13 CNMR (201 MHz, DMSO-d6): δ 169.47, 149.54, 145.17, 137.61, 137.39, 135.97, 133.99, 130.04, 129.80, 128.78, 126.39, 125.42, 123.88, 123.71, 122.26, 115.51, 43.08; ESI-MS (m / z): calcd. for C 18 H 13 F3N2O [M+H] + 331.10, found 331.25.

[0241] (42) Synthesis of the target compound 2-(3-chlorophenyl)-N-(quinolin-6-yl)acetamide (Code WRZ-18)

[0242]

[0243] Dissolve 300 mg (2.08 mmol) of 6-aminoquinoline in 30 mL of DCM, and add 505 mg (5 mmol) of triethylamine to obtain a quinoline solution. Dissolve 470 mg (2.5 mmol) of intermediate 4f in 10 mL of dichloromethane, and dropwise add the dichloromethane solution of intermediate 4f to the quinoline solution, and react overnight with stirring, and monitor the reaction by TLC. When the raw material is completely reacted, obtain the product 430 mg by silica gel column chromatography and recrystallization with a yield of 63%. 1H NMR (800 MHz, DMSO-d6): δ 10.55 (s, NH), 8.79 (d, J = 4.3 Hz, ArH), 8.39 (d, J = 2.7 Hz, ArH), 8.28 (d, J = 8.4 Hz, ArH), 7.98 (d, J = 9.0 Hz, 1H), 7.82 (dd, J = 9.0, 2.5 Hz, 1H), 7.49 - 7.46 (m, 2H), 7.38 (d, J = 7.8 Hz, 1H), 7.34 (d, J = 9.4 Hz, 2H), 3.77 (s, CH2); 13 C NMR (201 MHz, DMSO-d6): δ 169.48, 149.51, 145.14, 138.67, 137.42, 136.00, 133.31, 130.61, 130.00, 129.61, 128.78, 128.47, 127.09, 123.74, 122.26, 115.50, 43.14; ESI-MS (m / z): calcd. for C 17 H 13 ClN2O [M+H] + 298.06, found 298.12.

[0244] (43) Synthesis of the target compound 2-(3,4-difluorophenyl)-N-(quinolin-6- yl)acetamide (Code WRZ-19)

[0245]

[0246] Intermediate 4g was dissolved in 10 mL of dichloromethane, the dichloromethane solution of intermediate 4g was added dropwise to the quinoline solution, the reaction was carried out overnight, TLC was used to monitor the reaction, after the raw material was completely reacted, the product 208 mg was obtained by silica gel column chromatography and recrystallization, with a yield of 56%. 1H NMR (800 MHz, DMSO-d6): δ 10.53 (s, NH), 8.79 (d, J = 2.5 Hz, ArH), 8.38 (d, J = 2.6 Hz, ArH), 8.27 (d, J = 8.2 Hz, ArH), 7.98 (d, J = 9.1 Hz, 1H), 7.81 (dd, J = 9.1, 2.5 Hz, 1H), 7.49 - 7.38 (m, 3H), 7.21 (s, 1H), 3.76 (s, CH2); 13C NMR (201 MHz, DMSO-d6): δ 169.44, 150.14, 149.54, 148.25, 145.17, 137.40, 135.96, 133.88, 130.02, 128.77, 126.62, 123.72, 12; ESI-MS (m / z): calcd. for C17H12FN2O [M+H]+298.09, found 298.56.

[0247] (44) Synthesis of the target compound 2-(3,4-dimethoxyphenyl)-N-(quinolin-6-yl)acetamide (Code WRZ-20)

[0248]

[0249] Intermediate 4h was dissolved in 10 mL of dichloromethane, the dichloromethane solution of intermediate 4h was added dropwise to the quinoline solution, the reaction was carried out overnight under magnetic stirring, TLC was used to monitor the reaction, after the raw material was completely reacted, the product 206 mg was obtained by silica gel column chromatography and recrystallization, with a yield of 64%. 1H NMR (800 MHz, DMSO-d6): δ 10.45 (s, NH), 8.78 (d, J = 2.5 Hz, ArH), 8.38 (d, J = 2.4 Hz, ArH), 8.27 (d, J = 10.3 Hz, ArH), 7.97 (d, J = 9.0 Hz, 1H), 7.82 (dd, J = 9.0, 2.4 Hz, 1H), 7.47 (dd, J = 8.2, 4.1 Hz, 1H), 6.99 (s, 1H), 6.90 (q, J = 9.2 Hz, 2H), 3.76 (s, CH3), 3.73 (s, CH3), 3.64 (s, CH2); 13 CNMR (201 MHz, DMSO-d6): δ 170.33, 149.44, 149.02, 148.13, 145.10, 137.60, 135.96, 129.95, 128.78, 128.63, 123.77, 122.24, 121.58, 115.37, 113.54, 112.31, 56.01, 55.93, 43.42; ESI-MS (m / z): calcd. for C 19 H 18 N2O3[M+H] + 323.13, found 323.41.

[0250] (45) Synthesis of the target compound 2-bromo-2-(3-methoxyphenyl)-N- (quinolin-6-yl)acetamide (Code WRZ-21)

[0251]

[0252] Dissolve 560 mg (4.5 mmol) of 6-aminoquinoline in 30 mL of dichloromethane, and add 1.11 g (11.02 mmol) of triethylamine to obtain a quinoline solution. Dissolve 1.4 g (5.51 mmol) of intermediate 3a in 15 mL of dichloromethane, and dropwise add the dichloromethane solution of intermediate 3a to the quinoline solution, and react overnight, monitoring by TLC. After the reaction is completed, extract with dichloromethane, combine the organic phases, dry over anhydrous magnesium sulfate, and purify by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain 700 mg of the target compound, with a yield of 42%. 1H NMR (800 MHz, DMSO-d6): δ 10.55 (s, 1H), 8.78 (d, J = 4.2 Hz, 1H), 8.38 (s, 1H), 8.26 (d, J = 8.4 Hz, 1H), 7.98 (d, J = 9.0 Hz, 1H), 7.82 (d, J = 9.0 Hz, ArH), 7.53 - 7.46 (m, ArH), 7.08 (d, J = 3.4 Hz, ArH), 6.84 (dd, J = 8.7, 3.4 Hz, 1H), 3.90 (s, 2H), 3.77 (s, 3H); 13 C NMR (201 MHz, DMSO-d6): δ 170.62, 159.69, 144.99, 143.83, 140.07, 137.49, 135.74, 129.91, 129.82, 128.04, 122.81, 122.73, 121.89, 115.51, 115.28, 115.20, 112.52, 55.48, 43.76, 43.72; ESI-MS (m / z): calcd for C 18 H 15 BrN2O2[M+H] + 372.02, found 372.51.

[0253] (46) Synthesis of the target compound 2-bromo-2-(3-fluorophenyl)-N- (quinolin-6-yl)acetamide (Code WRZ-22)

[0254]

[0255] Dissolve 280 mg (2.25 mmol) of 6-aminoquinoline in 20 mL of dichloromethane, and add 550 mg (5.5 mmol) of triethylamine to obtain a quinoline solution. Dissolve 1.02 g (2.7 mmol) of intermediate 3b in 10 mL of dichloromethane, and dropwise add the dichloromethane solution of 3b to the quinoline solution, and react at room temperature for 8 hours with constant stirring, and monitor the reaction progress by TLC. After the reaction is completed, first filter out the insoluble matter, concentrate the filtrate, extract with dichloromethane, combine the organic phases, and remove water with anhydrous magnesium sulfate. After concentration, perform silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain 370 mg of the target compound, with a yield of 38.2%. 1HNMR (800 MHz, DMSO-d6): δ 10.67 (s, NH), 9.13 (d, J = 5.0 Hz, ArH), 8.95 (d, J = 8.5 Hz, ArH), 8.75 (s, ArH), 8.31 (d, J = 9.2 Hz, ArH), 8.19 (d, J = 9.2 Hz, 1H), 7.93 (dd, J = 8.5, 4.9 Hz, 1H), 7.43 (d, J = 4.2 Hz, 2H), 7.16 (d, J = 11.8 Hz, 1H), 5.31 (s, CH); 13 C NMR (201 MHz, DMSO-d6): δ 169.68, 149.57, 145.14, 138.87, 137.47, 136.00, 133.31, 130.61, 130.00, 129.61, 128.78, 128.41, 127.69, 123.44, 121.26, 115.56, 42.15; ESI-MS (m / z): calcd. for C 17 H 12 BrFN2O[M+H] + 360.01, found 360.88.

[0256] (47) Synthesis of the target compound 2-bromo-N-(quinolin-6-yl)-2-(m-tolyl)acetamide (Code WRZ-23)

[0257]

[0258] Dissolve 260 mg (1.86 mmol) of 6-aminoquinoline in 20 mL of dichloromethane, and add 453 mg (4.48 mmol) of triethylamine to obtain a quinoline solution. Dissolve 553 mg (2.24 mmol) of intermediate 3c in 10 mL of dichloromethane, and drop the dichloromethane solution of intermediate 3c into the quinoline solution, and react overnight for 8 hours, monitored by TLC (dichloromethane:methanol = 20:1). After the reaction is completed, first filter out the solid insoluble matter, concentrate the filtrate, extract with an appropriate amount of dichloromethane for 3 times, combine the organic phases, remove water in the organic phase with anhydrous magnesium sulfate, and chromatograph on a silica gel column to obtain 200 mg of the target compound, with a yield of 30%. 1HNMR (800 MHz, DMSO-d6): δ 10.59 (s, NH), 9.12 (d, J = 5.0 Hz, ArH), 8.92 (dd, J = 17.5, 8.5 Hz, ArH), 8.76 (s, ArH), 8.31 (d, J = 9.2 Hz, ArH), 8.19 (d, J = 9.4 Hz, 1H), 8.13 - 8.07 (m, 1H), 7.94 - 7.89 (m, 1H), 7.53 - 7.47 (m, 1H), 7.40 - 7.35 (m, 1H), 7.26 (t, J = 7.6 Hz, 1H), 7.12 (d, J = 7.5 Hz, 1H), 2.33 (d, J = 16.1 Hz, 3H); 13 C NMR (201 MHz, DMSO-d6): δ 169.48, 149.51, 145.14, 138.67, 137.42, 136.00, 133.31, 130.61, 130.00, 129.61, 128.78, 128.47, 127.09, 123.74, 122.26, 115.50, 43.14; ESI-MS (m / z): calcd for C 18 H 15 BrN2O[M+H] + 356.03 found 356.21.

[0259] (48) Synthesis of the target compound 2-bromo-2-(3,4-dichlorophenyl)-N- (quinolin-6-yl)acetamide (Code WRZ-24)

[0260]

[0261] Dissolve 330 mg (2.29 mmol) of 6-aminoquinoline in 25 mL of dichloromethane, and add 767 mg (7.6 mmol) of triethylamine, dissolve 827 mg (2.75 mmol) of intermediate 3d in 10 mL of dichloromethane, drop the dichloromethane solution of intermediate 3d into the quinoline solution, and react overnight for 8 hours, monitor by TLC (dichloromethane:methanol = 20:1), after the reaction is completed, first filter out the insoluble matter, concentrate the filtrate, extract with dichloromethane for 3 times, combine the organic phase, remove water with anhydrous magnesium sulfate, and chromatograph with silica gel column (dichloromethane:methanol = 300:1) to obtain 330 mg of the target compound, with a yield of 35%. 1H NMR (800 MHz, DMSO-d6): δ 10.70 (s, NH), 9.15 (d, J = 5.0 Hz, ArH), 8.98 (d, J = 8.6 Hz, ArH), 8.76 (s, ArH), 8.31 (d, J = 11.6 Hz, ArH), 8.21 (t, J = 9.1 Hz, 1H), 8.11 - 8.05 (m, 1H), 7.95 (dd, J = 8.5, 4.9 Hz, 1H), 7.83 (s, 1H), 7.67 (d, J = 8.5 Hz, 1H), 7.56 (d, J = 8.5 Hz, 1H); 13 C NMR (201 MHz, DMSO-d6): δ 169.21, 149.55, 145.18, 137.35, 135.97, 131.86, 131.22, 130.85, 130.25, 130.18, 129.82, 123.62, 122.27, 114.44, 48.27, 41.59; ESI-MS (m / z): calcd for C 17 H 11 BrCl2N2O [M+H] + 409.94 found 410.20.

[0262] (49) Synthesis of the target compound 2-bromo-N-(quinolin-6-yl)-2-(3- (trifluoromethyl)phenyl)acetamide (Code WRZ-25)

[0263]

[0264] Dissolve 320 mg (2.22 mmol) of 6-aminoquinoline in 20 mL of dichloromethane, and add 540 mg (5.38 mmol) of triethylamine to obtain a quinoline solution. Dissolve 800 mg (2.66 mmol) of intermediate 3e in 10 mL of dichloromethane, and drop the dichloromethane solution of intermediate 3e into the quinoline solution, and react overnight for 8 hours, TLC monitoring (dichloromethane:methanol = 20:1), after the raw material is completely reacted, first filter out the solid insoluble matter, concentrate the filtrate, extract with an appropriate amount of dichloromethane three times, combine the organic phases, remove water from the organic phase with anhydrous magnesium sulfate, and chromatograph on a silica gel column to obtain 360 mg of the target compound, with a yield of 40%. 1H NMR (800 MHz, DMSO-d6): δ 10.75 (s, NH), 9.15 (d, J = 4.9 Hz, ArH), 8.97 (d, J = 8.4 Hz, ArH), 8.77 (s, ArH), 8.32 (d, J = 9.4 Hz, ArH), 8.21 (t, J = 8.9 Hz, 1H), 7.96 - 7.93 (m, 2H), 7.90 (d, J = 7.8 Hz, 1H), 7.70 (d, J = 7.3 Hz, 1H), 7.65 (t, J = 7.8 Hz, 1H), 5.43 (s, 1H); 13 C NMR (201 MHz, DMSO-d6): δ 169.47, 149.54, 145.17, 137.61, 137.39, 135.97, 133.99, 130.04, 129.80, 128.78, 126.39, 125.42, 123.88, 123.71, 122.26, 115.51, 43.08; ESI-MS (m / z): calcd for C 18 H 12 BrF3N2O [M+H] + 410.01 found 410.25.

[0265] (50) Synthesis of the target compound 2-bromo-2-(3-chlorophenyl)-N- (quinolin-6-yl)acetamide (Code WRZ-26)

[0266]

[0267] Dissolve 380 mg (2.68 mmol) of 6-aminoquinoline in 20 mL of dichloromethane, and add 650 mg (6.44 mmol) of triethylamine to obtain a quinoline solution. Dissolve 760 mg (3.22 mmol) of intermediate 3f in 10 mL of dichloromethane, and dropwise add the dichloromethane solution of intermediate 3f to the quinoline solution, and react at room temperature for 8 hours, and monitor by TLC (dichloromethane:methanol = 30:1). After the reaction is completed, first filter off the insoluble matter, concentrate the filtrate, extract with dichloromethane, combine the organic phases, and dry over anhydrous magnesium sulfate. Subject to silica gel column chromatography (dichloromethane:methanol = 300:1) to obtain 340 mg of the target compound, with a yield of 35%. 1H NMR (800 MHz, DMSO-d6): δ 10.69 (s, 1H), 9.14 (d, J = 6.3 Hz, 1H), 8.95 (d, J = 8.7 Hz, 1H), 8.76 (s, 1H), 8.31 (d, J = 9.2 Hz, 1H), 8.20 (d, J = 9.2 Hz, 1H), 8.10 (dd, J = 15.2, 8.4 Hz, 1H), 7.95 - 7.89 (m, 1H), 7.65 (s, 1H), 7.55 (d, J = 7.5 Hz, 1H), 7.44 - 7.37 (m, 2H); 13 CNMR (201 MHz, DMSO-d6): δ 169.48, 149.51, 145.14, 138.67, 137.42, 136.00, 133.31, 130.61, 130.00, 129.61, 128.78, 128.47, 127.09, 123.74, 122.26, 115.50, 43.14; ESI-MS (m / z): calcd for C 17 H 12 BrClN2O [M+H] + 375.98, found 376.21.

[0268] (51) Synthesis of the target compound 2-bromo-2-(3,4-difluorophenyl)-N- (quinolin-6-yl)acetamide (Code WRZ-27)

[0269]

[0270] Dissolve 330 mg (2.2 9 mmol) of 6-aminoquinoline in 25 mL of dichloromethane, and add 767 mg (7.6 mmol) of triethylamine to obtain a quinoline solution. Dissolve 687 mg (2.75 mmol) of intermediate 3g in 10 mL of dichloromethane, and dropwise add the dichloromethane solution of intermediate 3g to the quinoline solution, and react overnight for 8 hours, monitoring by TLC (dichloromethane:methanol = 20:1). After the reaction is completed, first filter off the insoluble matter, concentrate the filtrate, extract with dichloromethane, combine the organic phases, and dry over anhydrous magnesium sulfate. Subject to silica gel column chromatography (dichloromethane:methanol = 300:1) to obtain 430 mg of the target compound. 1H NMR (800 MHz, DMSO-d6): δ 10.67 (s, NH), 9.13 (d, J = 5.0 Hz, ArH), 8.95 (d, J = 8.6 Hz, ArH), 8.75 (s, ArH), 8.30 (d, J = 9.2 Hz, ArH), 8.19 (d, J = 9.1 Hz, 1H), 8.10 (t, J = 10.8 Hz, 1H), 7.96 - 7.92 (m, 1H), 7.62 (t, J = 9.7 Hz, 1H), 7.49 - 7.38 (m, 2H); 13 C NMR (201 MHz, DMSO-d6): δ 169.35, 150.20, 149.54, 148.92, 148.25, 145.17, 137.29, 135.96, 133.86, 130.02, 128.61, 126.59, 123.72, 122.26, 118.81, 117.70, 115.51, 42.45; ESI-MS (m / z): calcd for C 17 H 11 BrF2N2O [M+H] + 378.00, found 378.36.

[0271] The compound of formula III can be prepared according to the following reaction scheme:

[0272]

[0273] wherein the reagents used are: (d) Triphosgene, Et3N, DCM, 0 °C; (e) Et3N, primary amines, DCM, rt, 8 h; (f) 30% H2O2, K2CO3, DMSO, 0 °C→ rt; 2 h; (g) NH2OH.HCl, KOH, MeOH / DCM, rt, 3-6 h.

[0274] Synthesis of intermediate 4-isocyanatobenzoic acid methyl ester (6a code 4)

[0275]

[0276] Dissolve 453 mg (3 mmol) of methyl 4-aminobenzoate in 20 mL of dichloromethane and add 910 mg (9 mmol) of triethylamine to obtain a solution of methyl 4-aminobenzoate. Dissolve 445 mg (1.5 mmol) of triphosgene in 10 mL of dichloromethane and slowly drop the triphosgene solution into the solution of methyl 4-aminobenzoate at 0 °C (under ice bath). React for 1 h under ice bath. TLC monitors the reaction to be complete. Concentrate the reaction solution under vacuum, wash with EA, and precipitate a large amount of salt. Filter the salt, reserve the filtrate, and concentrate under vacuum to obtain an orange oil, which is directly used in the next step.

[0277] (53) Synthesis of intermediate: p-cyanophenyl isocyanate (Code 6b)

[0278]

[0279] The synthesis method of intermediate 6b is the same as that of intermediate 6a. Dissolve 435 mg (3.68 mmol) of 4-aminobenzonitrile in 20 mL of dichloromethane and add 1.1 g (11.04 mmol) of triethylamine to obtain a solution of 4-aminobenzonitrile. Dissolve 545 mg (1.84 mmol) of triphosgene in 15 mL of dichloromethane (ultrasonic dissolution), and slowly drop the triphosgene solution into the solution of 4-aminobenzonitrile at 0 °C (under ice bath). React for 1 h under ice bath. TLC monitors the reaction to be complete. Concentrate the reaction solution under vacuum, wash with EA, and filter the salt under suction. Concentrate the filtrate under vacuum to obtain an orange oil, which is directly used in the next step.

[0280]

[0281] 370 mg (2.1 mmol) of intermediate 6a, 260 mg (1.8 mmol) of 5-aminoisoquinoline, and 354 mg (3.5 mmol) of triethylamine. React overnight, and purify by silica gel column chromatography using dichloromethane:methanol = 100:1 as the mobile phase to obtain 400 mg of white solid with a yield of 70%. 1 H NMR (400 MHz, DMSO-d6): δ 9.48 (s, NH), 9.33 (s, NH), 9.01 (s, ArH), 8.61 (d, J = 6.0 Hz, 1H), 8.29 (d, J = 7.6 Hz, 1H), 7.96 (dd, J = 14.1, 7.3 Hz, 3H), 7.86 (d, J = 8.1 Hz, 1H), 7.67 (t, J = 8.6 Hz, 3H), 3.84 (s, 3H); 13C NMR (201 MHz, DMSO-d6): δ 166.41, 153.18, 152.95, 144.64, 143.10, 133.82, 130.98, 129.16, 128.78, 128.01, 123.16, 121.60, 117.87, 114.97, 52.29. ESI-MS (m / z): calcd for C 18 H 15 N3O3[M+H] + 322.14, found 322.27.

[0282] (55) Synthesis of intermediate: 1-(4-cyanophenyl)-3-(isoquinolin-5-yl)urea (Code 7b)

[0283]

[0284] The synthesis method of intermediate 7b is the same as that of intermediate 7a. 696 mg (4.84 mmol) of 5-aminoisoquinoline was dissolved in 30 mL of dichloromethane, and 980 mg (9.68 mmol) of triethylamine was added to the solution under stirring to obtain a 5-aminoisoquinoline solution. 832 mg (5.8 mmol) of p-cyanophenyl isocyanate was dissolved in 30 mL of dichloromethane, and then the isocyanate solution was slowly added to the 5-aminoisoquinoline solution, and the reaction was carried out at room temperature for 8 hours. TLC monitoring was performed, and after the raw material was completely reacted, the product was insoluble in dichloromethane, and the solid product was obtained by suction filtration. No further purification was required, and the product was directly used in the next step. The by-product did not affect the synthesis of the next step.

[0285] (56) Synthesis of target compound: N-hydroxy-4-(3-(isoquinolin-5-yl)ureido)benzamide (Code WRZ-28)

[0286]

[0287] 430 mg (1.34 mmol) of intermediate 7a, 2.37 g (42.8 mmol) of potassium hydroxide, 2.88 g (30.9 mmol) of hydroxylamine hydrochloride, solvent: a mixture of dichloromethane 30 mL and methanol 15 mL (MeOH / DCM = 2:1), stirring at room temperature for 6 h. White solid 190 mg was obtained, with a yield of 44%. 1H NMR (400 MHz, DMSO-d6): δ 11.13 (s, NH), 9.34 (d, J = 10.9 Hz, NH), 8.99 (d, J = 18.7 Hz, ArH), 8.60 (d, J = 5.4 Hz, 1H), 8.29 (d, J = 7.4 Hz, 1H), 7.98 (d, J = 5.0 Hz, 1H), 7.85 (d, J = 7.9 Hz, 1H), 7.75 (d, J = 8.1 Hz, 2H), 7.67 (t, J = 7.8 Hz, 1H), 7.59 (d, J = 8.1 Hz, 2H); 13 C NMR (101 MHz, DMSO-d6): δ 164.55, 153.23, 153.05, 143.18, 142.72, 134.00, 129.18, 128.62, 128.41, 127.99, 126.49, 122.88, 121.23, 117.84, 114.90; ESI-MS (m / z): calcd. for C 17 H 14 N4O3[M+H] + 323.11, found 323.57.

[0288] (57) Synthesis of the target compound: 4-(3-(isoquinolin-5-yl)ureido)benzamide (Code WRZ-29)

[0289]

[0290] To a solution of 560 mg (1.94 mmol) of intermediate 7b in 25 mL of dimethyl sulfoxide, 1.37 g (9.9 mmol) of potassium carbonate was added with stirring, then 7.7 mL of 30% hydrogen peroxide was slowly added dropwise. 362 mg of white solid was obtained, yield 61%. 1 HNMR (400 MHz, DMSO-d6): δ 11.13 (s, NH), 9.34 (d, J = 10.9 Hz, NH), 8.99 (d, J = 18.7 Hz, ArH), 8.60 (d, J = 5.4 Hz, 1H), 8.29 (d, J = 7.4 Hz, 1H), 7.98 (d, J = 5.0 Hz, 1H), 7.85 (d, J = 7.9 Hz, 1H), 7.75 (d, J = 8.1 Hz, 2H), 7.67 (t, J = 7.8 Hz, 1H), 7.59 (d, J = 8.1 Hz, 2H); 13 C NMR (101 MHz, DMSO-d6): δ 164.55, 153.23, 153.05, 143.18, 142.72, 134.00, 129.18, 128.62, 128.41, 127.99, 126.49, 122.88, 121.23, 117.84, 114.90; ESI-MS (m / z): calcd. for C17 H 14 N4O2[M+H] + 307.11, found 307.84.

[0291] Example 2 Inhibition activity of compounds on ROCK2

[0292] The ADP-Glo Luminescence Assay was used to test the inhibition activity of the target compounds on ROCK2. ADP-Glo is a detection method that reflects the activity of kinase by detecting the fluorescence intensity. The kinase activity is reflected by detecting the content of ADP directly generated in the kinase reaction. The detection method can be divided into three steps. In the first step, ATP is converted to ADP under the action of kinase, and polypeptide is phosphorylated at the same time. In the second step, the ADP-Glo reagent degrades the excess ATP in the system. In the third step, under the action of the kinase detection reagent, ADP is reduced to ATP, and ATP is transferred to luciferin under the action of Ultra-GloTM luciferase, and the kinase activity is detected by the fluorescence signal generated. The stronger the fluorescence signal, the stronger the kinase activity, and the positive correlation. The ADP-Glo luminescence assay has obvious advantages. It can detect the activity of ADP generating enzyme without the need for antibodies and radioactive labels.

[0293] (1) Preparation of kinase reaction buffer: 1x kinase buffer 40mM Tris, pH 7.5 20mM MgCl2; 1mM DTT 0.10% BSA

[0294] (2) Preparation of compounds for detecting kinase:

[0295] 1) Dilute the compound to 100 times the final concentration of the reaction with 100% DMSO. For example, if the desired highest inhibitor concentration is 10 μM, prepare a 1000 μM compound DMSO solution in this step.

[0296] 2) Transfer 40 μL of compound dilution to a 384-well plate.

[0297] 3) In the same 384-well plate, add 40 μL of 100% DMSO to 2 empty wells, no compound control and no enzyme control. Label the plate as the source plate.

[0298] 4) Transfer 50 nL per well from the 384-well echo plate to the 384-well detection plate by echo method.

[0299] (3) Kinase reaction:

[0300] 1) Add 2.5 μL of 2x enzyme solution to each well of the 384-well detection plate (add 2.5 μL of 1x kinase buffer to each well of the 384-well detection plate except for the low control well).

[0301] 2) Add fam labeled peptide and ATP in 1x kinase base buffer.

[0302] 3) Add 2.5 μL of 2x peptide solution to each well of a 384 well assay plate.

[0303] 4) Incubate the 384 well assay plate at 28°C for 60 minutes.

[0304] 5) Add 5 μL of ADP-Glo reagent 1 and incubate at 28°C for 180 minutes.

[0305] 6) Add 10 μL of ADP-Glo reagent 2 and incubate at room temperature for 30 minutes.

[0306] (4) Experimental results processing: 1) Copy the RLU values from the Envision program.

[0307] 2) Convert the RLU values to percent inhibition values, Inhibition = (max - sample RLU) / (max - min)*100.

[0308] Where "min" is the RLU of the no enzyme control and "max" is the RLU of the DMSO control.

[0309] The results of the inhibition of ROCK2 enzyme by the compounds prepared in Example 1 are shown in Table 1 below:

[0310]

[0311]

[0312]

[0313] According to the results in Table 1, at a concentration of 10 μM, the inhibition rates of WRZ-1, WRZ-2, WRZ-3, WRZ-4, WRZ-5, WRZ-6, WRZ-7, WRZ-8, WRZ-9, WRZ-14, WRZ-15, WRZ-21, WRZ-22, WRZ-24, WRZ-28, and WRZ-29 of the above-mentioned compounds on ROCK2 enzyme are all more than 50%, indicating that the above-mentioned compounds have good enzyme inhibition activity; further, the inhibition rates of compounds WRZ-1, WRZ-2, WRZ-3, WRZ-4, WRZ-6, WRZ-7, WRZ-8, WRZ-9, WRZ-15, WRZ-21, and WRZ-22 are more than 90%, which is comparable to the positive drug Fasudil.

[0314] Example 3 In vivo vasodilation activity experiment of the compound

[0315] Animals: 5-6 weeks old male C57BL / 6 mice, 20-25 g. The animal feeding conditions are room temperature 20-25℃, humidity 45%-60%, free feeding and drinking water.

[0316] Reagents and instruments: phenylephrine (PE), sodium chloride (NaCl), potassium chloride (KCl), potassium dihydrogen phosphate (KH2PO4), magnesium sulfate (MgSO4), sodium bicarbonate (NaHCO3), D-Glucose (C6H 12 O6), calcium chloride (CaCl2), dimethyl sulfoxide (DMSO), microsurgical dissecting tools, DMT vascular tension measurement system, pipette, electronic balance, constant temperature water bath, optical microscope. (PE is dissolved in pure water, and the compound is dissolved in DMSO).

[0317] Tension measurement of isolated blood vessels (preparation of vascular rings):

[0318] Krebs solution composition: NaCl 118mM, KCl 4.7mM, MgSO4 1.2mM, KH2PO4 1.2mM, NaHCO3 25mM, CaCl2 1.3mM, D-Glucose 10mM.

[0319] 60mmol / L K + -Kerbs buffer composition: NaCl 22.7mM, KCl 100mM, MgSO4 1.2mM, KH2PO4 1.2mM, NaHCO3 25mM, EGTA 1mM, D-Glucose 10mM.

[0320] After the male C57BL / 6 mice were executed by decapitation, the thoracic aorta was quickly removed and placed in pre-cooled 4℃ Krebs filled with pure oxygen. The surrounding connective tissue and adipose tissue were removed, and endothelial intact vascular ring specimens 2-3mm long were prepared. The vascular ring specimens were connected to the DMT vascular tension measurement system by two thin steel wires and hung horizontally in a 5mL constant temperature bath. The PowerLab data acquisition and analysis system was used to record the changes in vascular tension. Pure oxygen was continuously introduced into the constant temperature bath, 5mL of Krebs solution was added, and the temperature was maintained at 37℃. Zero was adjusted to the point where there was no significant change in vascular tension, a 3mN pre-tension was given, and the pre-tension was adjusted to be stable. The vascular ring specimens were equilibrated in 37℃, 100% O2 saturated Kerbs solution for 60min (Kerbs solution was replaced every 20min during this period). 60mmol / L K + -Kerbs buffer repeatedly stimulated the vascular ring specimens twice, and the difference in contraction amplitude of the two stimulations was less than 10%, indicating that the vascular activity was good, and the next experimental operation could be performed.

[0321] Relaxation activity test:

[0322] After the vessel rings were fully equilibrated, endothelium-intact C57BL / 6 mouse thoracic aortic rings were pre-constricted with PE (1 x 10 -6 mol / L) or 60 mmol / L K + -Kerbs buffer, and after reaching a stable plateau, a cumulative dose of compound was added, and the vasorelaxation rate was calculated. Vasorelaxation rate (%) = (tension of the vessel ring after administration of the compound - maximum contraction tension of the vessel ring induced by PE or 60 mmol / L K + -Kerbs buffer) / (tension of the vessel ring induced by PE or 60 mmol / L K + -Kerbs buffer - basal tension of the vessel ring) x 100%. The statistical data were plotted as the vasorelaxation rate (%) versus the logarithm of the compound concentration (logc).

[0323] Experimental results and analysis: Figures 1 to 3 The vasorelaxation activity results of some compounds in mouse aortic rings exposed to 1 x 10 -6 mol / L of the alpha-adrenergic receptor agonist phenylephrine (Phe) or 60 mM KCl to induce contraction are shown. Among them, WRZ-1, WRZ-13, and WRZ-6 can effectively relax the vessel contraction induced by Phe or KCl in the concentration range of 3 μM to 300 μM. WRZ-1 can relax the vessel contraction induced by Phe in the administered concentration range (3-100 μM), and the maximum relaxation rate is 74.47%. WRZ-1 can relax the vessel contraction induced by KCl in the administered concentration range (3-100 μM), (n = 3), and the maximum relaxation rate is 73.26%. WRZ-13 can relax the vessel contraction induced by Phe in the administered concentration range (3-300 μM), (n = 3), and the maximum relaxation rate is 77.55%. WRZ-13 can relax the vessel contraction induced by KCl in the administered concentration range (3-300 μM), (n = 3), and the maximum relaxation rate is 95.29%. WRZ-6 can relax the vessel contraction induced by Phe in the administered concentration range (3-300 μM), (n = 3), and the maximum relaxation rate is 82.99%. WRZ-6 can relax the vessel contraction induced by KCl in the administered concentration range (3-300 μM), (n = 3), and the maximum relaxation rate is 82.72%.

[0324] Example 4 In vivo mouse hypertension model activity detection results of compound WRZ-6

[0325] Experimental procedure: 6-8 weeks C57BL / 6 mice, under the condition of light / dark cycle 12 hours, room temperature 20-25℃, humidity 40-60%, after 5 days of adaptive feeding, 20 healthy male mice with body weight of 20-25 grams were selected for experiment. 5 mice were taken as normal group by random number table method, without modeling, the remaining 15 mice were intraperitoneally injected with N'-nitro-L-arginine (L-NNA) solution (1 mg / mL) at 15 mg / kg, 2 times / d, for 21 days. The tail artery blood pressure of mice during modeling was measured once a week. The systolic pressure ≥160 mmHg (1 mmHg=0.133 kPa) was regarded as the standard of successful modeling. After successful modeling, 15 model mice were randomly divided into model group (5 mice), WRZ-6 group (5 mice) and captopril group (5 mice). The captopril group was given captopril solution (7.615 mg / kg, adult clinical equivalent dose) by gavage, the WRZ-6 group was given WRZ-6 suspension (7.615 mg / kg) by gavage, and the normal group and the model group were given the same amount of normal saline by gavage. The drug volume was 10 mL / kg, and the drug administration lasted for 4 weeks. The tail artery blood pressure of mice during modeling was measured once a week. The specific drug dosage is shown in the following table:

[0326] Table of drug dosage for hypertension model

[0327]

[0328] Measurement of mouse tail artery blood pressure: the systolic pressure (SBP) and diastolic pressure (DBP) of each group of mice were measured before and after drug administration every week. Before the measurement of basic blood pressure, each mouse was habituated for at least 5 consecutive days. The blood pressure measurement should be carried out in the designated quiet area. Turn on the heating device of the non-invasive blood pressure detector, set the temperature to 37℃. The measuring mouse was fixed in the fixator, preheated for 5 min, and the pulse sleeve was put on the 1 / 3 position of the mouse tail close to the heart. Start the detector system, the inductor will automatically apply pressure for blood pressure measurement. Each mouse was measured three times repeatedly, and then the average value was taken.

[0329] Experimental results and analysis: The high blood pressure mouse model was established by intraperitoneal injection of N'-nitro-L-arginine (L-NNA). After 21 days of modeling, the systolic blood pressure (SBP) was ≥160 mm Hg (1 mm Hg = 0.133 kPa), which proved that the modeling was successful. WRZ-6 and positive drug Captopril were used for gavage treatment. After administration, the systolic blood pressure (SBP) and diastolic blood pressure (DBP) of the WRZ-6 group and the positive drug Captopril group continued to decrease, which proved that WRZ-6 had good effect on treating high blood pressure (n = 5) Figure 4

[0330] Example 5 Docking study of the compound with ROCK2

[0331] Experimental principle: Discovery Studio is a computer molecular simulation software developed by Tripos Company in the United States. DS LibDock molecular docking module is a fast molecular docking tool, which is suitable for fast and accurate virtual screening of large-scale databases. LibDock matches the conformation of small molecules with the hot spot of the receptor according to the principle of interaction, and docks the conformation into the binding pocket of the receptor. Its biggest advantage is fast, can operate in parallel, and is suitable for large-scale virtual screening, which can provide theoretical support and technical guidance for discovering new ROCK2 target lead compounds and structural modification and modification.

[0332] Experimental steps:

[0333] 1. Define the protein receptor

[0334] (1) Open the protein in DS: File→open, add hydrogen: Chemistry→hydrogens→add;

[0335] (2) Define the protein receptor: Tools→Receptor-Ligand Interactions→Define and Edit Binding Site→Define Receptor 2H9V. Ctrl+h calls up the tree display area, where SBD_receptor appears.

[0336] (3) Define the binding site: Define site→from receptor cavities; Adjust the size of the binding site: change site size→expand or contract.

[0337] 2. Define ligand

[0338] ​(1) Open ligand in DS: File→open, add hydrogen: Chemistry→hydrogens→add;

[0339] (2) Prepare ligand: Tools→Small Molecules→Prepare or Filter Ligands (Prepare Ligands);

[0340] (3) Search small molecule conformations, generate 9 conformations WRZ-7(1);

[0341] (4) Minimization→Full Minimization, wherein Input ligands select WRZ-7-(1):All;

[0342] →Run, end display poses optimized.

[0343] 3. Molecular docking

[0344] Tools→Receptor-Ligand Interactions→Dock Ligands→Dock Ligands (LibDock), wherein Input Receptor selects 2H9V:2H9V, Input Ligands selects WRZ-7-(1):All, Input site sphere is the sphere region defined previously; click Run to run molecular docking.

[0345] Result analysis: As shown in Figure 5 the docking results show that the designed compound WRZ-7 can also very well dock in the active pocket, and at the same time, the key amino acids MET-172 and GLU-170 in the hinge region form hydrogen bond interactions, and the bromine atom forms halogen bond interactions with the amino acid residue LYS-121, which may be the key to the good drug activity of the halogen-substituted derivatives. The D region at the top of the docking pocket forms interactions with the amino acid residue PHE-103. This greatly increases the affinity of the compound with the target. These results show the scientific rationality of the designed compound of the present application.

[0346] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. For those skilled in the art, the present application can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A compound, characterized in that it is... 2-bromo- N - (1 H - indazol-5-yl) -2- (3-methoxyphenyl) acetamide.

2. A pharmaceutical composition, characterized by, The compound of claim 1, a pharmaceutically acceptable salt of the compound.

3. A pharmaceutical preparation, characterized in that, A pharmaceutical composition comprising an active ingredient which is a compound of claim 1 or a pharmaceutical composition of claim 2 and a pharmaceutically acceptable carrier.

4. Use of a compound of claim 1, a pharmaceutical composition of claim 2 or a pharmaceutical preparation of claim 3 for the manufacture of a ROCK inhibitor.

5. The use according to claim 4, wherein the compound is ###0002### The ROCK inhibitor is used for the manufacture of a model agent for a ROCK inhibition model.

6. The use according to claim 4, wherein the compound is ###0002### The ROCK inhibitor is used for the manufacture of a model agent for preventing, ameliorating or treating a disease associated with high expression of ROCK.

7. The use according to claim 5, wherein the compound is ###0002### The model is a cell model, a tissue model or an animal model.

8. The use according to claim 4, wherein the compound is ###0002### The ROCK inhibitor is a ROCK2 inhibitor.

9. Use of a compound of claim 1, a pharmaceutical composition of claim 2 or a pharmaceutical preparation of claim 3 for the manufacture of a vasodilator or a blood pressure lowering drug.

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