CN113980003A - 2- ((2-methoxyphenyl) sulfonyl) isoindoline compound and preparation method thereof - Google Patents

Abstract

The invention relates to the technical field of medicaments, and provides a 2- ((2-methoxyphenyl) sulfonyl) isoindoline compound and a preparation method thereof. The 2- ((2-methoxyphenyl) sulfonyl) isoindoline compound is a compound with a structure shown in a formula I or a pharmaceutically acceptable salt thereof. The structure of the formula I is as follows:

wherein R is₁Is methoxy, fluoro, chloro, bromo, cyano, aminocarbonyl, acetyl, dimethylamino, trifluoromethyl, R₂1H-indazol-5-yl and 1H-pyrazol-4-yl. The 2- ((2-methoxyphenyl) sulfonyl) isoindoline compound can be used as an effective ROCK inhibitor. The pharmaceutical composition containing the compound also has good pharmacological activity for treating tumors, cardiovascular diseases, nervous system diseases and fibrotic diseases.

Description

2- ((2-methoxyphenyl) sulfonyl) isoindoline compound and preparation method thereof

Technical Field

The invention relates to the technical field of medicines, in particular to a 2- ((2-methoxyphenyl) sulfonyl) isoindoline compound and a preparation method thereof.

Background

Rho belongs to small molecule single polymer GTPase superfamily, is a mammalian gene homolog of RAS superfamily, and regulates the recombination of a cell actin skeleton through a downstream most main effector Rho kinase, thereby widely participating in a series of biological processes such as cell mitosis, cytoskeleton regulation, nerve regeneration, tumor cell infiltration, apoptosis and the like. Rho kinase has now been found to be predominantly of two subtypes: ROCK1, the former being mainly present in non-neural tissues such as heart, lung, etc., and ROCK2, the latter being mainly present in the central nervous system such as neurons, cerebral cortex, etc.

Rho is abnormally activated in a variety of cardiovascular diseases, such as atherosclerosis, hypertension, pulmonary hypertension, and the like. Rho inhibitors exhibit benefits in the treatment of these diseases in animal models. In addition, there is evidence that inhibition of Rho kinase activation in vivo also has the effects of promoting neuronal synapse growth and promoting recovery of neurological function following injury, and can be used for treating and alleviating central nervous diseases such as spinal cord injury, Alzheimer's disease, and neuroinflammation, spinal cord demyelination. In addition, tumor infiltration and migration are also dependent on Rho kinase activation, inhibition of Rho kinase activation also inhibits tumor metastasis.

The only ROCK inhibitors currently on the market are erl (for the treatment of cerebral vasospasm) and Glanatec (for the treatment of ocular hypertension and glaucoma). Therefore, the development of the ROCK inhibitor with novel structure and strong activity has important significance.

Disclosure of Invention

In order to search for a novel ROCK inhibitor, the invention designs and synthesizes a series of 2- ((2-methoxyphenyl) sulfonyl) isoindoline compounds which have novel structures and high Rho kinase inhibition activity through extensive and intensive research, and the ROCK inhibition activity of the compounds is researched.

The invention aims to provide a 2- ((2-methoxyphenyl) sulfonyl) isoindoline compound or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a process for the preparation of the above compound.

It is another object of the present invention to provide a pharmaceutical composition comprising the above compound.

Another object of the present invention is to provide the use of the above compounds and the above pharmaceutical compositions as Rho kinase inhibitors for the treatment of tumors, cardiovascular diseases, neurological diseases, fibrotic diseases.

In order to achieve the purpose, the invention adopts the technical scheme that:

in a first aspect, the present invention provides a compound having the structure of formula i:

wherein R is₁Is methoxy, fluorine, chlorine, bromine, cyano, aminocarbonyl, acetyl, dimethylamino, trifluoromethyl,

R₂1H-indazol-5-yl and 1H-pyrazol-4-yl.

The compound with the structure of formula I or the pharmaceutically acceptable salt thereof provided by the invention can be selected from the compounds with the structure of the following compounds:

。

in a second aspect, the present invention provides a process for the preparation of a compound having the structure of formula i or a pharmaceutically acceptable salt thereof, comprising the steps of:

and (3) synthesizing an intermediate IV:

reacting a compound II with a structure shown in a formula II and a compound III with a structure shown in a formula III in a first solvent in the presence of a first base at a first reaction temperature to generate an intermediate IV with a structure shown in a formula IV;

synthesis of Compound I:

and (3) carrying out coupling reaction on the intermediate IV and the compound V with the structure of the formula V in a second solvent in the presence of a second base under the action of a catalyst at a second reaction temperature to generate the compound with the structure of the formula I.

As a preferable embodiment of the preparation method provided by the present invention, the first solvent is at least one of tetrahydrofuran, dioxane, toluene, N-dimethylformamide, dichloromethane, and acetonitrile;

and/or the second solvent is any one of dioxane, N-dimethylformamide, toluene, dioxane/water and glycol dimethyl ether.

As another preferable embodiment of the preparation method provided by the present invention, the first reaction temperature is 20 to 100 ℃;

and/or the second reaction temperature is 40-120 ℃.

As another preferable embodiment of the preparation method provided by the present invention, the first base is at least one of triethylamine, N-diisopropylethylamine, N-methylmorpholine, potassium carbonate, cesium carbonate, and sodium carbonate;

and/or the second base is at least one of cesium carbonate, sodium tert-butoxide, potassium carbonate, sodium acetate and potassium phosphate.

As another preferable embodiment of the preparation method provided by the invention, the catalyst is tetratriphenylPalladium phosphine (Pd (PPh)₃)₄) Palladium acetate (Pd (OAc)₂) Tris (dibenzylideneacetone) dipalladium (Pd)₂(dba)₃) 1,1' -bis (diphenylphosphino) ferrocene dichloropalladium (Pd) (dppf) Cl₂) At least one of

The preparation method provided by the invention takes a compound II and a compound III as initial raw materials, firstly synthesizes an intermediate IV, and then carries out coupling reaction on the intermediate IV and a compound V to prepare the compound with the structure of a formula I. The whole preparation process is simple to operate, easy to control, low in requirements on production equipment and suitable for industrial large-scale production.

In a third aspect, the present invention provides a pharmaceutical composition comprising a compound having the structure of formula i, or a pharmaceutically acceptable salt thereof.

In the present invention, a compound having the structure of formula i or a pharmaceutically acceptable salt thereof is a pharmaceutically active ingredient in a pharmaceutical composition.

The present invention provides pharmaceutical compositions comprising, in addition to a therapeutically effective amount of at least one compound having the structure of formula i:

one or more pharmaceutical excipients;

and/or one or more pharmaceutically active substances having ROCK inhibitory activity other than the compound having the structure of formula i or a pharmaceutically acceptable salt thereof.

Methods of preparing the pharmaceutical compositions provided herein will be apparent to those skilled in the art and include conventional mixing, dissolving, lyophilizing and like techniques.

The pharmaceutical composition provided by the invention can be prepared into various common dosage forms, such as tablets, pills, capsules, granules, oral solutions, oral suspensions, oral emulsions, injections and the like, according to conventional preparation methods in the pharmaceutical field. It is convenient to provide the patient with clinical use by administering to the patient by various common modes of administration, such as oral or parenteral administration (by intravenous, intramuscular, topical or subcutaneous routes).

In a fourth aspect, the invention provides a compound having a structure shown in formula I, a pharmaceutically acceptable salt thereof and application of a pharmaceutical composition containing the compound having the structure shown in formula I and the pharmaceutically acceptable salt thereof as a ROCK inhibitor in preparation of medicines for treating tumors, cardiovascular diseases, nervous system diseases and fibrotic diseases.

Further, the tumor is selected from:

skin cancer, bladder cancer, ovarian cancer, breast cancer, stomach cancer, prostate cancer, colon cancer, lung cancer, bone cancer, brain cancer, rectal cancer, esophageal cancer, tongue cancer, kidney cancer, cervical cancer, corpus uteri cancer, testicular cancer, urinary cancer, melanoma, astrocytoma, meningioma, hodgkin's lymphoma, non-hodgkin's lymphoma, acute lymphatic leukemia, chronic lymphatic leukemia, acute myeloid leukemia, chronic myeloid leukemia, adult T-cell leukemia lymphoma, hepatocellular carcinoma, multiple myeloma, basal cell tumor, seminoma, chondrosarcoma, myosarcoma, fibrosarcoma.

Some of the terms involved in the expression of the present invention are defined as follows:

the term "pharmaceutically acceptable salts" refers to those salts that retain the biological effectiveness and properties of the parent compound. The salt comprises:

acid addition salts obtained by reaction of the free base of the parent compound with an inorganic acid or with an organic acid; the inorganic acid comprises hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, phosphoric acid, sulfuric acid, perchloric acid and the like; the organic acid includes acetic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, fumaric acid, gluconic acid, glutamic acid, isethionic acid, lactic acid, mandelic acid, mucic acid, pamoic acid, pantothenic acid, succinic acid, malonic acid, or the like; preferably hydrochloric acid or (L) -malic acid;

or, a salt formed when an acid proton present in the parent compound is replaced with a metal ion or coordinated with an organic base; the metal ions include alkali metal ions, alkaline earth ions, aluminum ions and the like; the organic base includes ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, etc.

The term "hydrate" refers to a substance in which water molecules are coordinately or covalently bound to cations or anions in a compound, or to a substance in which water ions are not directly bound to cations or anions but exist at certain positions in a solid crystal lattice in a certain proportion.

The term "solvate" refers to a substance formed by association of a compound with molecules of a pharmaceutically acceptable solvent, which typically includes ethanol, acetic acid, and the like.

The term "stereoisomer" refers to compounds having the same molecular formula, but differing in the arrangement of the atoms in the molecule in space. Since the compounds provided by the present invention may have one or more asymmetric centers, the compounds may be prepared as (R) -stereoisomers alone or (S) -stereoisomers alone or as mixtures thereof. Unless otherwise indicated, the description or designation of a particular compound in the present invention is intended to include the individual enantiomers as well as racemic or other mixtures thereof. Methods for determining stereochemical configuration and separating stereoisomers are routine in the art (see the discussion in chapter 4 of Ad V attached Organic Chemistry, 4 th edition, J.March, John Wiley and Sons, New York, 1992). Thus, the present invention also encompasses any stereoisomeric form, its corresponding enantiomers (d-and l-or (+) and (-) isomers) and diastereomers thereof and mixtures thereof, having the ability to modulate RET kinase activity, and is not limited to any one stereoisomeric form.

The term "pharmaceutical composition" refers to a mixture of one or more of the compounds provided herein with other chemical ingredients (e.g., pharmaceutical excipients). The purpose of the pharmaceutical composition is to facilitate the administration of the compound to the organism.

The term "pharmaceutical excipient" refers to a substance that has been reasonably evaluated in terms of safety, does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the compound administered, and is included in a pharmaceutical preparation, in addition to an active ingredient. The pharmaceutic adjuvant has important functions of solubilization, dissolution assistance, sustained and controlled release and the like besides excipient, carrier and stability improvement, and is an important component which may influence the quality, safety and effectiveness of the pharmaceutical composition. The pharmaceutical excipients include, but are not limited to, carriers, diluents, excipients, solubilizers, binders, disintegrants, penetration enhancers, pH adjusters, buffers, release retardants, flavoring agents, preservatives, antioxidants, and the like.

The invention has the beneficial effects that:

(1) the invention provides a 2- ((2-methoxyphenyl) sulfonyl) isoindoline compound or a pharmaceutically acceptable salt thereof, which can be used as an effective ROCK inhibitor and has stronger inhibitory activity.

(2) The invention also provides a pharmaceutical composition containing the 2- ((2-methoxyphenyl) sulfonyl) isoindoline compound or pharmaceutically acceptable salt thereof, which has good pharmacological activity for treating tumors, cardiovascular diseases, nervous system diseases and fibrotic diseases.

Detailed Description

In order to make the technical problems, technical solutions and advantageous effects to be solved by the present invention more apparent, the present invention is further described in detail below with reference to specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the present patent and do not limit the scope of the invention in any way.

In the preparation method of the compound with the structure of formula I or the pharmaceutically acceptable salt thereof, the specific reaction process is as follows:

in the first step, intermediate IV is synthesized: dissolving a compound II, a compound III and a first base in a first solvent, and reacting at 20-100 ℃ to generate an intermediate IV.

Wherein the first base can be at least one selected from triethylamine, N-diisopropylethylamine, N-methylmorpholine, potassium carbonate, cesium carbonate and sodium carbonate.

The first solvent may be at least one selected from tetrahydrofuran, dioxane, toluene, N-dimethylformamide, dichloromethane, and acetonitrile.

For example, 1mol of compound II, 1mol of compound III and 1.5mol of N, N-diisopropylethylamine are dissolved in 5L of tetrahydrofuran and the reaction is stirred at 40 ℃ to give intermediate IV. The reaction can be monitored by Thin Layer Chromatography (abbreviated as TLC), and after the reaction is finished, the intermediate iv can be obtained by conventional processing techniques such as extraction, drying, concentration, separation and the like.

The second step is the synthesis of a compound having the structure of formula i: and dissolving the intermediate IV, the compound V, a second alkali and a catalyst in a second solvent, and performing coupling reaction at 40-120 ℃ to generate the compound with the structure of the formula I.

Wherein the second base is at least one selected from cesium carbonate, sodium tert-butoxide, potassium carbonate, sodium acetate and potassium phosphate.

The second solvent can be any one of dioxane, N-dimethylformamide, toluene, dioxane/water and ethylene glycol dimethyl ether.

The catalyst can be selected from palladium tetratriphenylphosphine (Pd (PPh)₃)₄) Palladium acetate (Pd (OAc)₂) Tris (dibenzylideneacetone) dipalladium (Pd)₂(dba)₃) 1,1' -bis (diphenylphosphino) ferrocene dichloropalladium (Pd) (dppf) Cl₂) At least one of (1).

For example, 1mol of intermediate IV, 1mol of compound V, 2mol of sodium acetate and 0.1mol of palladium acetate (Pd (OAc)₂) Dissolving in 4L dioxane, and stirring at 75 deg.C to obtain the compound with structure shown in formula I. The reaction can be monitored by TLC, and after the reaction is finished, the final product can be obtained by conventional treatment processes such as concentration, separation and the like.

The invention is described in more detail by referring to a part of the tests, which are carried out in sequence, and the following detailed description is given by combining specific examples:

unless defined otherwise, technical terms used in the following examples have the same meanings as commonly understood by one of ordinary skill in the art to which the present invention belongs. The reagents used in the following examples, unless otherwise specified, are all conventional biochemical reagents; the raw materials, instruments, equipment and the like used in the following examples are either commercially available or available by existing methods; the dosage of the reagent is the dosage of the reagent in the conventional experiment operation if no special description exists; the experimental methods are conventional methods unless otherwise specified.

Example 1

5-methoxy-2- ((2-methoxy-4- (1H-pyrazol-4-yl) phenyl) sulfonyl) isoindoline

The first step is as follows:

dissolving compound 1a (2.84 g, 10.0 mmol), compound 1b (1.5 g, 10.0 mmol) and N, N-Diisopropylethylamine (DIEA) (1.9 g, 15.0 mmol) in dichloromethane (50 mL), reacting at room temperature, monitoring the reaction by TLC, adding water (50 mL) after the reaction is finished, quenching the reaction, drying the organic layer, filtering, concentrating, and separating by column chromatography to obtain 2.6g of light yellow solid (compound 1 c), wherein the yield is 65.5%;

the second step is that:

compound 1c (397 mg, 1.0 mmol), compound 1d (112 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), palladium tetrakistriphenylphosphine (Pd (PPh)₃)₄) (115 mg, 0.1 mmol) was dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃ and monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL. times.2), the organic layer was concentrated, and column chromatography was performed to give 224mg of off-white solid (Compound 1) in 58.2% yield. ESI (+) m/z = 386.1.

Example 2

5-fluoro-2- ((2-methoxy-4- (1H-pyrazol-4-yl) phenyl) sulfonyl) isoindoline

The first step is as follows:

dissolving compound 1a (2.84 g, 10.0 mmol), compound 2a (1.37 g, 10.0 mmol) and DIEA (1.9 g, 15.0 mmol) in dichloromethane (50 mL), reacting at room temperature, monitoring the reaction by TLC, adding water (50 mL) after the reaction to quench the reaction, drying the organic layer, filtering, concentrating, and separating by column chromatography to obtain a light yellow solid (compound 2 b) 2.7g with a yield of 70.3%;

the second step is that:

compound 2b (385 mg, 1.0 mmol), compound 1d (112 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, the reaction was monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography was performed to obtain 257mg of off-white solid (compound 2), with a yield of 68.9%. ESI (+) m/z = 374.1.

Example 3

5-chloro-2- ((2-methoxy-4- (1H-pyrazol-4-yl) phenyl) sulfonyl) isoindoline

The first step is as follows:

dissolving compound 1a (2.84 g, 10.0 mmol), compound 3a (1.5 g, 10.0 mmol) and DIEA (1.9 g, 15.0 mmol) in dichloromethane (50 mL), reacting at room temperature, monitoring the reaction by TLC, adding water (50 mL) after the reaction to quench the reaction, drying the organic layer, filtering, concentrating, and separating by column chromatography to obtain a light yellow solid (compound 3 b) 2.2g with a yield of 55.0%;

the second step is that:

compound 3b (400 mg, 1.0 mmol), compound 1d (112 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, the reaction was monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography was performed to obtain 209mg of off-white solid (compound 3), with a yield of 53.6%. ESI (+) m/z = 390.1.

Example 4

5-dimethylamino-2- ((2-methoxy-4- (1H-pyrazol-4-yl) phenyl) sulfonyl) isoindoline

The first step is as follows:

dissolving compound 1a (2.84 g, 10.0 mmol), compound 4a (1.6 g, 10.0 mmol) and DIEA (1.9 g, 15.0 mmol) in dichloromethane (50 mL), reacting at room temperature, monitoring the reaction by TLC, adding water (50 mL) after the reaction to quench the reaction, drying the organic layer, filtering, concentrating, and separating by column chromatography to obtain a light yellow solid (compound 4 b) 1.9g with a yield of 46.3%;

the second step is that:

compound 4b (410 mg, 1.0 mmol), compound 1d (112 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, the reaction was monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography was performed to obtain an off-white solid (compound 4) 252mg, with a yield of 63.3%. ESI (+) m/z = 399.1.

Example 5

5-acetyl-2- ((2-methoxy-4- (1H-pyrazol-4-yl) phenyl) sulfonyl) isoindoline

The first step is as follows:

dissolving compound 1a (2.84 g, 10.0 mmol), compound 5a (1.6 g, 10.0 mmol) and DIEA (1.9 g, 15.0 mmol) in dichloromethane (50 mL), reacting at room temperature, monitoring the reaction by TLC, adding water (50 mL) after the reaction to quench the reaction, drying the organic layer, filtering, concentrating, and separating by column chromatography to obtain a light yellow solid (compound 5 b) 2.5g with a yield of 61.1%;

the second step is that:

compound 5b (409 mg, 1.0 mmol), compound 1d (112 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, the reaction was monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography was performed to obtain 271mg of off-white solid (compound 5) with a yield of 68.3%. ESI (+) m/z = 398.1.

Example 6

5-aminocarbonyl-2- ((2-methoxy-4- (1H-pyrazol-4-yl) phenyl) sulfonyl) isoindoline

The first step is as follows:

dissolving compound 1a (2.84 g, 10.0 mmol), compound 6a (1.6 g, 10.0 mmol) and DIEA (1.9 g, 15.0 mmol) in dichloromethane (50 mL), reacting at room temperature, monitoring the reaction by TLC, adding water (50 mL) after the reaction to quench the reaction, drying the organic layer, filtering, concentrating, and separating by column chromatography to obtain a light yellow solid (compound 6 b) 2.2g with a yield of 53.8%;

the second step is that:

compound 6b (409 mg, 1.0 mmol), compound 1d (112 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, the reaction was monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography was performed to obtain 285mg of a white solid (compound 6), with a yield of 71.6%. ESI (+) m/z = 399.1.

Example 7

5-trifluoromethyl-2- ((2-methoxy-4- (1H-pyrazol-4-yl) phenyl) sulfonyl) isoindoline

The first step is as follows:

dissolving compound 1a (2.84 g, 10.0 mmol), compound 7a (1.9 g, 10.0 mmol) and DIEA (1.9 g, 15.0 mmol) in dichloromethane (50 mL), reacting at room temperature, monitoring the reaction by TLC, adding water (50 mL) after the reaction to quench the reaction, drying the organic layer, filtering, concentrating, and separating by column chromatography to obtain a light yellow solid (compound 7 b) 2.6g with a yield of 59.8%;

the second step is that:

compound 7b (434 mg, 1.0 mmol), compound 1d (112 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, the reaction was monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography was performed to obtain 225mg of a white solid (compound 7), with a yield of 53.2%. ESI (+) m/z = 424.1.

Example 8

4-methoxy-2- ((2-methoxy-4- (1H-pyrazol-4-yl) phenyl) sulfonyl) isoindoline

The first step is as follows:

dissolving compound 1a (2.84 g, 10.0 mmol), compound 8a (1.5 g, 10.0 mmol) and DIEA (1.9 g, 15.0 mmol) in dichloromethane (50 mL), reacting at room temperature, monitoring the reaction by TLC, adding water (50 mL) after the reaction to quench the reaction, drying the organic layer, filtering, concentrating, and separating by column chromatography to obtain a light yellow solid (compound 8 b) 2.8g with a yield of 70.5%;

the second step is that:

compound 8b (397 mg, 1.0 mmol), compound 1d (112 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, monitored by TLC, extracted with ethyl acetate (50 mL × 2) after completion of the reaction, the organic layer was concentrated, and column chromatography gave 248mg of a white solid (compound 8) in 64.4% yield. ESI (+) m/z = 386.1.

Example 9

4-aminocarbonyl-2- ((2-methoxy-4- (1H-pyrazol-4-yl) phenyl) sulfonyl) isoindoline

The first step is as follows:

dissolving compound 1a (2.84 g, 10.0 mmol), compound 9a (1.6 g, 10.0 mmol) and DIEA (1.9 g, 15.0 mmol) in dichloromethane (50 mL), reacting at room temperature, monitoring the reaction by TLC, adding water (50 mL) after the reaction to quench the reaction, drying the organic layer, filtering, concentrating, and separating by column chromatography to obtain a light yellow solid (compound 9 b) 2.3g with a yield of 56.1%;

the second step is that:

compound 9b (410 mg, 1.0 mmol), compound 1d (112 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, the reaction was monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography was performed to obtain 208mg of a white solid (compound 9), with a yield of 52.3%. ESI (+) m/z = 399.1.

Example 10

4-fluoro-2- ((2-methoxy-4- (1H-pyrazol-4-yl) phenyl) sulfonyl) isoindoline

The first step is as follows:

dissolving compound 1a (2.84 g, 10.0 mmol), compound 10a (1.4 g, 10.0 mmol) and DIEA (1.9 g, 15.0 mmol) in dichloromethane (50 mL), reacting at room temperature, monitoring the reaction by TLC, adding water (50 mL) after the reaction to quench the reaction, drying the organic layer, filtering, concentrating, and separating by column chromatography to obtain a light yellow solid (compound 10 b) 2.0g with a yield of 51.9%;

the second step is that:

compound 10b (385 mg, 1.0 mmol), compound 1d (112 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, the reaction was monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography was performed to obtain 226mg of a white solid (compound 10), with a yield of 60.6%. ESI (+) m/z = 374.1.

Example 11

5- (3-methoxy-4- ((5-methoxyisoindolin-2-yl) sulfonyl) phenyl) -1H-indazole

Compound 1c was synthesized according to the procedure of the first step in example 1.

Compound 1c (397 mg, 1.0 mmol), compound 11a (162 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), tetrakistriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, monitored by TLC, after completion of the reaction, extracted with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography gave 255mg of white solid (compound 11) in 58.6% yield, ESI (+) m/z = 436.1.

Example 12

5- (3-methoxy-4- ((5-fluoroisoindolin-2-yl) sulfonyl) phenyl) -1H-indazole

Compound 2b was synthesized according to the procedure of the first step in example 2.

Compound 2b (385 mg, 1.0 mmol), compound 11a (162 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, the reaction was monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography was performed to obtain 278mg of a white solid (compound 12) with a yield of 65.7%. ESI (+) m/z = 424.1.

Example 13

5- (3-methoxy-4- ((5-chloroisoindolin-2-yl) sulfonyl) phenyl) -1H-indazole

Compound 3b was synthesized according to the procedure of the first step in example 3.

Compound 3b (400 mg, 1.0 mmol), compound 11a (162 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, the reaction was monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography was performed to obtain 278mg of a white solid (compound 13) in a yield of 63.3%. ESI (+) m/z = 440.1.

Example 14

5- (3-methoxy-4- ((5-dimethylaminoindolin-2-yl) sulfonyl) phenyl) -1H-indazole

Compound 4b was synthesized according to the procedure of the first step in example 4.

Compound 4b (410 mg, 1.0 mmol), compound 11a (162 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, the reaction was monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography was performed to obtain 240mg of a white solid (compound 14), with a yield of 53.6%. ESI (+) m/z = 449.2.

Example 15

5- (3-methoxy-4- ((5-acetylisoindolin-2-yl) sulfonyl) phenyl) -1H-indazole

Compound 5b was synthesized according to the procedure of the first step in example 5.

Compound 5b (409 mg, 1.0 mmol), compound 11a (162 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, the reaction was monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography was performed to obtain 254mg of a white solid (compound 15) with a yield of 56.8%. ESI (+) m/z = 448.1.

Example 16

5- (3-methoxy-4- ((5-aminocarbonylisoindolin-2-yl) sulfonyl) phenyl) -1H-indazole

Compound 6b was synthesized according to the procedure of the first step in example 6.

Compound 6b (409 mg, 1.0 mmol), compound 11a (162 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, the reaction was monitored by TLC, after completion of the reaction, extraction was performed with ethyl acetate (50 mL × 2), the organic layer was concentrated, and column chromatography was performed to obtain 282mg of a white solid (compound 16) with a yield of 62.8%. ESI (+) m/z = 449.1.

Example 17

5- (3-methoxy-4- ((5-trifluoromethyl isoindolin-2-yl) sulfonyl) phenyl) -1H-indazole

Compound 7b was synthesized according to the procedure of the first step in example 7.

Compound 7b (434 mg, 1.0 mmol), compound 11a (162 mg, 1.0 mmol), potassium carbonate (276 mg, 2.0 mmol), and tetratriphenylphosphine palladium (115 mg, 0.1 mmol) were dissolved in dioxane (30 mL) and water (10 mL), the reaction was stirred at 80 ℃, monitored by TLC, extracted with ethyl acetate (50 mL × 2) after completion of the reaction, the organic layer was concentrated, and column chromatography gave 269mg of a white solid (compound 17) in 56.9% yield. ESI (+) m/z = 474.1.

Biological evaluation

In this example, GSK269962 is an internal control compound by detecting the inhibition effect of the compounds 1 to 17 prepared in the examples 1 to 17 on ROCK1(ROCK2) kinase.

The experimental method comprises the following steps:

diluting the compound with dimethyl sulfoxide (DMSO) to 10 different concentrations for use;

buffer solution: 40mM Tris pH 7.5, 20mM MgCl2, 0.1% BSA, 50. mu.M DTT;

adding 10 μ L of 2.5 × 0.1 μ g/mL ROCK1(ROCK2) working solution into a 96-well plate;

adding 5 μ L of 5 × compound solution to a 96-well plate, mixing, and incubating at 25 deg.C for 10 min;

add 10. mu.L 2.5X 37.5. mu.g/mL S6K substrate and 12.5. mu.M ATP mix working solution, incubate 60 minutes at 30 ℃;

adding 25 μ L of the reaction mixture to another 96-well plate, adding 25 μ L of ADP-Glo reagent, mixing, incubating at 25 ℃ for 60 min, and terminating the reaction;

adding 40 mul of the mixed solution for terminating the reaction into 40 mul of the kinase detection reagent, mixing uniformly, and incubating for 40 minutes at 25 ℃;

reading Iummescence signal value, and calculating to obtain compound IC₅₀. The test results were as follows:

as can be seen from the above table, compounds 1 to 17 all have certain inhibitory effects on ROCK1(ROCK2) kinase, wherein the IC of compound 1, compound 2, compound 3, compound 5, compound 6, compound 8, compound 9, compound 10, compound 11, compound 12, compound 16 and compound 17 on ROCK1₅₀Values less than 10 nM; IC of Compound 2, Compound 3, Compound 5, Compounds 11 to 15 against ROCK2₅₀Value less than 10nM。

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.

Claims (10)

1. A compound having the structure of formula I:

wherein R is₁Is methoxy, fluorine, chlorine, bromine, cyano, aminocarbonyl, acetyl, dimethylamino, trifluoromethyl,

R₂1H-indazol-5-yl and 1H-pyrazol-4-yl.

2. A process for the preparation of a compound having the structure of formula i, or a pharmaceutically acceptable salt thereof, as claimed in claim 1, comprising the steps of:

and (3) synthesizing an intermediate IV:

reacting a compound II with a structure shown in a formula II and a compound III with a structure shown in a formula III in a first solvent in the presence of a first base at a first reaction temperature to generate an intermediate IV with a structure shown in a formula IV;

synthesis of Compound I:

and (3) carrying out coupling reaction on the intermediate IV and the compound V with the structure of the formula V in a second solvent in the presence of a second base under the action of a catalyst at a second reaction temperature to generate the compound with the structure of the formula I.

3. The method according to claim 2, wherein the first solvent is at least one of tetrahydrofuran, dioxane, toluene, N-dimethylformamide, dichloromethane, and acetonitrile;

and/or the second solvent is any one of dioxane, N-dimethylformamide, toluene, dioxane/water and glycol dimethyl ether.

4. The method of claim 2, wherein the first reaction temperature is 20 to 100 ℃;

and/or the second reaction temperature is 40-120 ℃.

5. The method of claim 2, wherein the first base is at least one of triethylamine, N-diisopropylethylamine, N-methylmorpholine, potassium carbonate, cesium carbonate, and sodium carbonate;

and/or the second base is at least one of cesium carbonate, sodium tert-butoxide, potassium carbonate, sodium acetate and potassium phosphate.

6. The method according to claim 2, wherein the catalyst is at least one of tetrakistriphenylphosphine palladium, palladium acetate, tris-dibenzylideneacetone dipalladium, and 1,1' -bis (diphenylphosphino) ferrocene dichloropalladium.

7. A pharmaceutical composition comprising a compound having the structure of formula i as claimed in claim 1 or a pharmaceutically acceptable salt thereof.

8. Use of a compound having the structure of formula i or a pharmaceutically acceptable salt thereof according to claim 1 or a pharmaceutical composition according to claim 7 as a ROCK inhibitor in the manufacture of a medicament for the treatment of a tumor, a cardiovascular disease, a neurological disease, a fibrotic disease.

9. The use according to claim 8, wherein the tumor is selected from the group consisting of:

skin cancer, bladder cancer, ovarian cancer, breast cancer, stomach cancer, prostate cancer, colon cancer, lung cancer, bone cancer, brain cancer, rectal cancer, esophageal cancer, tongue cancer, kidney cancer, cervical cancer, corpus uteri cancer, testicular cancer, urinary cancer, melanoma, astrocytoma, meningioma, hodgkin's lymphoma, non-hodgkin's lymphoma, acute lymphatic leukemia, chronic lymphatic leukemia, acute myeloid leukemia, chronic myeloid leukemia, adult T-cell leukemia lymphoma, hepatocellular carcinoma, multiple myeloma, basal cell tumor, seminoma, chondrosarcoma, myosarcoma, fibrosarcoma.

10. The compound of claim 1 having the structure of formula i or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of compounds having the structure:

。