Benzothiophene derivative as well as preparation method and application thereof

Abstract

The invention discloses a benzothiophene derivative as well as a preparation method and application thereof, and relates to the technical field of medicines. The invention provides a benzothiophene derivative with a novel structure. The benzothiophene derivative has ROCK kinase inhibition activity and ROCK / HDAC double-target inhibition activity. Experiments prove that the compounds provided by the invention all show relatively high activity, and the prepared compounds have good ROCK / HDAC inhibitory activity, and particularly show relatively good activity in the aspects of breast cancer cell proliferation resistance and immunity. In addition, the invention also provides a method for preparing the benzothiophene derivative, and the method has the advantages of short synthetic route, simple preparation process and easy operation, and can meet the needs of large-scale industrial production.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a benzothiophene derivative and a preparation method and application thereof. Background Art

[0002] Rho-associated coiled-coil kinases (ROCKs) are a class of serine / threonine protein kinases belonging to the AGC kinase family and are downstream effector molecules of Rho small GTP-binding proteins. ROCK proteins contain an N-terminal kinase domain, a Rho protein binding domain (RBD), and a PH domain bisected by a zinc finger-like motif domain (CRD) at the C-terminus. In the classic ROCK signaling pathway, Rho GTPase activates downstream ROCK kinases and further phosphorylates ROCK downstream substrates, thereby remodeling the cytoskeleton, inducing actin-myosin contraction, and regulating microtubule dynamics.

[0003] ROCK kinase consists of two subtypes, ROCK1 and ROCK2, which have 65% homology in amino acid sequence and 92% similarity in the kinase domain. ROCK is widely distributed in tissues throughout the body, with ROCK1 being more highly expressed in non-neural tissues such as blood, small intestine and thymus, while ROCK2 is more highly expressed in the brain, heart and colon.

[0004] ROCK kinase is involved in the occurrence and development of many diseases, including eye diseases, cardiovascular and cerebrovascular diseases, tumor metastasis, central nervous system diseases and diabetic nephropathy, and has become an important target for the treatment of many diseases. To date, four ROCK inhibitors have successfully passed clinical trials and are on the market: Fasudil, Ripasudil, Netarsudil and KD025. In addition, the ROCK inhibitor Y27632 is currently undergoing a Phase I clinical trial for the treatment of corneal edema, and AT13148 has completed a Phase I study for the treatment of advanced solid tumors. However, due to the narrow therapeutic index and pharmacokinetic properties of AT13148, its development has been discontinued.

[0005] Histone deacetylases (HDACs) are an important class of enzymes that regulate chromatin structure and gene expression by removing acetyl groups from histones. HDACs belong to a large family that includes multiple subclasses and members, and they play a key role in various biological processes of cells. HDACs play a role in a variety of biological processes such as cell cycle, cell differentiation, apoptosis, DNA repair, and cell metabolism. They affect the fate and function of cells by regulating the expression of specific genes. They are divided into four categories based on sequence homology: class I HDAC (HDAC1, HDAC2, HDAC3, HDAC8), class IIa HDAC (HDAC4, HDAC5, HDAC7, HDAC9), class IIb HDAC (HDAC6, HDAC10), class III HDAC (Sirtuins), and class IV HDAC (HDAC11).

[0006] To date, many HDAC inhibitors have been developed, of which six have been approved for clinical use: Vorinostat (SAHA), Belinostat, Panobinostat, Pracinostat, Chidamide, and Romidepsin. However, HDAC inhibitors have limited therapeutic applications as single drugs, and past studies have often combined HDAC inhibitors with other protein inhibitors to form dual-target inhibitors to enhance therapeutic effects.

[0007] The interaction between ROCK and HDAC in various pathological processes provides new therapeutic strategies for the treatment of various diseases. These enzymes play a key role in cancer, inflammation, autoimmune diseases, fibrosis, transplant rejection, diseases related to specific cytokines such as IL-17, IL-21 and IL-23, as well as eye, cardiovascular and cerebrovascular, central nervous system, diabetic nephropathy and skin diseases. They jointly affect the occurrence and development of diseases by regulating cell behavior, inflammatory response and immune response. As an innovative treatment strategy, dual-target inhibitors provide a more effective treatment method for diseases by acting on two key enzymes, ROCK and HDAC, at the same time. This treatment method can target multiple links in the development of the disease, thereby improving the efficacy, while reducing the risk of side effects and drug resistance that may be caused by single-target treatment. Summary of the invention

[0008] Based on the deficiencies in the prior art, the applicant has designed a benzothiophene derivative or a pharmaceutically acceptable salt thereof having ROCK kinase inhibitory activity and a ROCK / HDAC dual-target inhibitory activity, as well as a preparation method and application of the benzothiophene derivative or a pharmaceutically acceptable salt thereof.

[0009] The present invention provides a benzothiophene derivative, the structure of which is shown in Formula I:

[0010]

[0011] wherein the R1 group is at least one of a hydrogen atom substituted at one position on the benzene ring, a halogen group, a pyrazolyl group, a pyridyl group, an isoquinolyl group and an indolyl group,

[0012] The R2 group is at least one of the following groups: hydrogen atom, halogen group, benzyl group, 5-7 membered aryl group, heteroaryl group, cycloalkyl group, heterocyclic group, alkyl group with 1 to 6 carbon atoms, haloalkyl group, haloalkoxy group, alkylhydroxamic acid, hydroxyalkyl group, cyano group, hydroxyl group, nitro group, carboxyl group.

[0013] Preferably, the R1 group is a pyrazolyl, pyridyl, isoquinolyl or indolyl substituted at one position on the benzene ring (positions 4 to 7 of the benzothiophene ring),

[0014] The R2 group is a 5- to 7-membered aryl group or an alkyl group having 1 to 6 carbon atoms.

[0015] Specifically, the benzothiophene derivative is any one of the following compounds:

[0016]

[0017]

[0018]

[0019] Further preferably, the benzothiophene derivative is any one of the following compounds:

[0020]

[0021] The experimental results in the specific embodiments of the present invention show that compounds 24, 27, 28, and 34 have better anti-MDA-MB-231 breast cancer cell proliferation activity than positive drugs SAHA and Y27632. Compound 24 exhibits more efficient ROCK1 and 2 kinase inhibitory activity. Compound 24 has better anti-tumor immune effect than positive drug SAHA.

[0022] The present invention provides a method for preparing the benzothiophene derivatives, comprising the following steps:

[0023] (1) A halogenated benzothiophene-2-carboxylic acid and an amine having a corresponding R2 group are subjected to an amidation reaction under the action of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate to obtain product 1;

[0024] (2) product 1 undergoes a Suzuki reaction (also known as a Suzuki coupling reaction or a Suzuki-Miyaura reaction) with a boronic acid having a corresponding R1 group to obtain the benzothiophene derivative;

[0025] The R1 group is a pyrazolyl, pyridyl, isoquinolyl or indolyl substituted at one position on the benzene ring,

[0026] The R2 group is a 5- to 7-membered aryl group.

[0027] Preferably, in step (1), the molar ratio of the halogenated benzothiophene-2-carboxylic acid to the amine having the corresponding R2 group is 1:1.5;

[0028] During the reaction, a catalyst is also added, and the catalyst is N-methylimidazole;

[0029] In step (2), product 1 and a boric acid having a corresponding R1 group are dissolved in a solvent, and then a catalyst and a base are added to react.

[0030] The molar ratio of the product 1 to the boronic acid having the corresponding R1 group is 1:2;

[0031] The solvent is a mixed solution of dioxane and water,

[0032] The catalyst is tetrakis(triphenylphosphine)palladium,

[0033] The base is potassium carbonate,

[0034] In the present invention, the conditions of the Suzuki reaction are: reacting at a reaction temperature of 100° C. for 16-20 hours under nitrogen protection.

[0035] The present invention also provides a method for preparing the benzothiophene derivatives, comprising the following steps:

[0036] (1) subjecting a halogenated benzothiophene-2-carboxylic acid and an ester having a corresponding R2 group to an amidation reaction under the action of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate to obtain product 1;

[0037] (2) subjecting product 1 to a deprotection reaction to obtain product 2;

[0038] (3) subjecting product 2 to an amidation reaction with O-2-tetrahydro-2H-pyranylhydroxylamine in the presence of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate to obtain product 3;

[0039] (4) Product 3 undergoes Suzuki reaction with a boronic acid having a corresponding R1 group to obtain product 4;

[0040] (5) subjecting the product 4 to a deprotection reaction to obtain the benzothiophene derivative;

[0041] The R1 group is a pyrazolyl or pyridyl group substituted at one position on the benzene ring,

[0042] The R2 group is an alkyl group having 1 to 6 carbon atoms.

[0043] Preferably, in step (1), the molar ratio of the halogenated benzothiophene-2-carboxylic acid to the ester having the corresponding R2 group is 1:1.5;

[0044] During the reaction, a catalyst is also added, and the catalyst is N-methylimidazole;

[0045] In steps (2) and (5), the deprotection reaction is carried out by dissolving product 1 or product 4 in dichloromethane and adding trifluoroacetic acid in an equal volume to dichloromethane;

[0046] In step (3), the molar ratio of product 2 to O-2-tetrahydro-2H-pyranylhydroxylamine is 1:1.5;

[0047] In step (4), the product 3 and the boric acid having the corresponding R1 group are dissolved in a solvent, and then a catalyst and a base are added to react.

[0048] The molar ratio of the product 3 to the boronic acid having the corresponding R1 group is 1:2;

[0049] The solvent is a mixed solution of dioxane and water,

[0050] The catalyst is tetrakis(triphenylphosphine)palladium,

[0051] The base is potassium carbonate,

[0052] In the present invention, the conditions of the Suzuki reaction are: reacting at a reaction temperature of 100° C. for 16-20 hours under nitrogen protection.

[0053] The present invention also provides the use of the benzothiophene derivatives in preparing drugs for treating and / or preventing diseases associated with abnormal activity of ROCK and / or HDAC.

[0054] Specifically, the disease associated with abnormal activity of ROCK and / or HDAC is selected from at least one of cancer, inflammatory diseases, autoimmune diseases, fibrotic diseases, transplant rejection, diseases associated with excessive secretion of IL-17, IL-21 and / or IL-23, eye diseases, cardiovascular and cerebrovascular diseases, central nervous system diseases, diabetic nephropathy and skin diseases.

[0055] More specifically, the diseases associated with abnormal activity of ROCK and / or HDAC are breast cancer, endometrial cancer, ovarian cancer, prostate cancer, colorectal cancer, lung cancer, rheumatoid arthritis or psoriasis. Inhibiting ROCK and / or HDAC in cancer can simultaneously inhibit the migration, invasion, proliferation and angiogenesis of cancer cells, thereby effectively inhibiting the growth and spread of tumors; in inflammatory diseases, it can reduce inflammatory reactions by inhibiting the migration of inflammatory cells and the expression of inflammatory genes; in autoimmune diseases, it can reduce the attack of the immune system on its own tissues by regulating the activity of immune cells and the expression of immune-related genes. Although the therapeutic effect of breast cancer is only shown in the embodiment, based on the mechanism of action of ROCK and HDAC in various diseases, it is speculated that the benzothiophene derivatives can also be used to treat other diseases associated with abnormal activity of ROCK and / or HDAC, such as endometrial cancer, ovarian cancer, prostate cancer, colorectal cancer, lung cancer, rheumatoid arthritis, psoriasis, etc.

[0056] The experimental results in the specific embodiments of the present invention show that compounds 24, 27, 28, and 34 have better anti-MDA-MB-231 breast cancer cell proliferation activity than positive drugs SAHA and Y27632. Compound 24 exhibits more efficient ROCK1 and 2 kinase inhibitory activity. Compound 24 has better anti-tumor immune effect than positive drug SAHA.

[0057] The present invention also provides a pharmaceutical composition comprising the benzothiophene derivative or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0058] The active compound can be prepared into a form suitable for administration by any appropriate route, and the composition of the present invention can be prepared by conventional methods using one or more pharmaceutically acceptable carriers. Therefore, the active compound of the present invention can be formulated into various dosage forms for oral administration, injection (e.g., intravenous, intramuscular or subcutaneous) administration, inhalation or insufflation administration. The compound of the present invention can also be formulated into dosage forms such as tablets, hard or soft capsules, aqueous or oily suspensions, emulsions, injections, dispersible powders or granules, suppositories, lozenges or syrups.

[0059] As a general guide, the active compound is preferably in a unit dose form, or in a single dose self-administered by a patient. The unit dose of the compound or composition of the present invention can be expressed in the form of tablets, capsules, cachets, bottled liquids, powders, granules, lozenges, suppositories, reconstituted powders or liquid preparations. Suitable unit doses can be 0.1 to 1000 mg.

[0060] The pharmaceutical composition of the present invention may contain one or more auxiliary materials in addition to the active compound, and the auxiliary materials are selected from the following ingredients: filler (diluent), binder, wetting agent, disintegrant or excipient, etc. Depending on the administration method, the composition may contain 0.1 to 99% by weight of the active compound.

[0061] As is well known to those skilled in the art, the dosage of a drug depends on a variety of factors, including but not limited to the following factors: the activity of the specific compound used, the severity of the disease, the age of the patient, the weight of the patient, the health status of the patient, the behavior of the patient, the diet of the patient, the time of administration, the mode of administration, the rate of excretion, the combination of drugs, etc.; in addition, the optimal treatment method such as the mode of treatment, the daily dosage or the type of pharmaceutically acceptable salt can be verified according to traditional treatment regimens.

[0062] The present invention has the following advantages and beneficial effects:

[0063] The present invention provides a class of novel benzothiophene derivatives, which have ROCK kinase inhibitory activity and ROCK / HDAC dual-target inhibitory activity. Experiments have shown that the compounds provided by the present invention all show high activity, and the prepared compounds have good ROCK / HDAC inhibitory activity, especially in anti-breast cancer cell proliferation and immunity. In addition, the present invention also proposes a method for preparing the benzothiophene derivatives, which has a short synthetic route, a simple preparation process, and is easy to operate, and can meet the needs of large-scale industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0064] Figure 1 The figure is a synthetic route diagram of benzothiophene derivatives of the present invention.

[0065] Figure 2 The B synthetic route of the benzothiophene derivatives of the present invention.

[0066] Figure 3 The effect of compound 24 and SAHA on T cells in tumors. All data are expressed as mean ± standard deviation. *p<0.05, **p<0.01, ***p<0.001. DETAILED DESCRIPTION

[0067] The present invention is further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. The experimental methods described in the following examples are conventional methods unless otherwise specified; the reagents and biological materials described are commercially available unless otherwise specified.

[0068] Example 1

[0069] The synthetic route of benzothiophene derivatives of the present invention is as follows Figure 1 As shown, the specific preparation method is as follows:

[0070] The first step is to synthesize 3a-d:

[0071] Accurately weigh halogenated benzothiophene-2-carboxylic acid (i.e. Figure 1 1a-d) (1.00 g, 3.89 mmol), m-methylbenzylamine (i.e. Figure 1 2a) (0.80 g, 5.84 mmol) and N-methylimidazole (1.12 g, 13.6 mmol) were mixed and dissolved in acetonitrile (20 mL). Then, N, N, N', N'-tetramethylchloroformamidine hexafluorophosphate (1.31 g, 4.67 mmol) was added at one time, and the mixture was stirred at room temperature for 24 hours. The mixture was cooled to room temperature, concentrated under reduced pressure to remove the solvent, and the residual liquid was extracted with dichloromethane, which was repeated three times and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with petroleum ether / ethyl acetate (1:1, v / v, 500 mL) to obtain compounds 3a-d with yields of 80%, 85%, 75%, and 80%, respectively.

[0072] Step 2: Synthesis of 4a-p(1-16):

[0073] The corresponding compounds 3a-d (0.50 mmol), compound 1H-pyrazole-4-boronic acid (i.e. Figure 1 X1 in (0.11 g, 1.00 mmol) or 4-pyridineboronic acid (i.e. Figure 1 X2 in (0.12 g, 1.00 mmol) or isoquinoline-5-boronic acid (i.e. Figure 1 X3 in (0.17 g, 1.00 mmol) or 1H-indole-5-boronic acid (i.e. Figure 1 A mixture of X4 (0.16 g, 1.00 mmol), tetrakis(triphenylphosphine)palladium (0.05 g, 0.04 mmol) and anhydrous potassium carbonate (0.35 g, 2.50 mmol) was stirred at 100 ° C for 20 hours under a nitrogen atmosphere. The solvent was a dioxane / water mixture (v / v = 4:1). Cooled to room temperature, concentrated under reduced pressure to remove the solvent, and the residual liquid was extracted with dichloromethane. The process was repeated three times and the organic phases were combined, dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure after filtration. The residue was purified by high performance liquid chromatography (separation conditions: preparative column Welch XB-C18, 21.2×250mm, 5μm; elution system: methanol / water (volume ratio) = 10%-100%, containing 0.5‰ trifluoroacetic acid by volume, flow rate: 10ml / min, elution time: 40min) to obtain the corresponding compounds 1-16 with yields of 86%, 86%, 85%, 76%, 86%, 86%, 80%, 78%, 88%, 86%, 85%, 76%, 86%, 85%, 86%, 76%, respectively.

[0074] Compound 1: 1 H NMR (600MHz, DMSO-d6) δ13.01 (s, 1H), 9.33 (t, J=6.2Hz, 1H), 8.16-8.14 (m, 2H), 8.10 (s, 1H), 7.99-7.97 (d, J=8.2Hz, 1H), 7.74-7.72 (dd, J= 8.2Hz, 1.4Hz, 1H), 7.63-7.52 (m, 1H), 7.25 (t, J=8.3Hz, 1H), 6.92-6.91 (m, 2H), 6.83-6.82 (m, 1H), 4.47-4.46 (d, J=6.2Hz, 2H), 3.73 (s, 3H). 13 C NMR (150MHz, DMSO-d6) δ161.62, 159.43, 140.96, 140.42, 139.99, 137.85, 130.11, 129.56, 129 .55, 124.83, 124.43, 123.17, 120.97, 120.82, 119.57, 119.56, 113.15, 112.31, 55.08, 42.72. ESI-MS m / z: 364.1119[M+H] + .

[0075] Compound 2: 1 H NMR (600MHz, DMSO-d6) δ8.92 (s, 1H), 8.74-8.73 (m, 2H), 8.36 (m, 1H), 8.17 (m, 1H), 8.15-8.13 (d, J=8.5Hz, 1H), 7.94-7.92 (m, 2H), 7.88-7.86 (m, 1H), 7.25 (t, J=8.5Hz, 1H), 6.96-6.95 (m, 2H), 6.84-6.82 (m, 1H), 4.52-4.51 (d, J=6.2Hz, 2H), 3.77 (s, 3H). 13C NMR (150MHz, DMSO-d6) δ160.96, 159.23, 147.68, 147.35, 141.15, 141.06, 140.82, 140.24, 139.44, 133 .18, 128.71, 124.63, 124.21, 123.06, 123.02, 121.51, 121.44, 119.23, 113.11, 112.13, 54.69, 42.54. ESI-MS m / z: 375.1162[M+H] + .

[0076] Compound 3: 1 H NMR (600MHz, DMSO-d6) δ9.70 (s, 1H), 9.37 (t, J=6.2Hz, 1H), 8.56-8.55 (d, J=6.3Hz , 1H), 8.40-8.39 (d, J=8.2Hz, 1H), 8.23-8.21 (m, 2H), 8.09 (s, 1H), 8.03-8.02 (d, J= 7.2Hz, 1H), 7.97-7.94 (m, 2H), 7.60-7.59 (dd, J=8.2Hz, 1.7Hz, 1H), 7.27-7.24 (m, 1 H), 6.93-6.92 (m, 2H), 6.83-6.81 (m, 1H), 4.49-4.48 (d, J=6.0Hz, 2H), 3.74 (s, 3H). 13 CNMR (150MHz, DMSO-d6) δ161.49, 159.44, 150.49, 141.20, 140.90, 140.14, 139.74, 138.57, 137.97, 134.91, 134.59, 13 4.35, 129.55, 128.99, 128.96, 128.20, 127.95, 126.34, 125.09, 123.35, 120.58, 119.59, 113.21, 112.31, 55.09, 42.77. ESI-MS m / z: 425.1329[M+H] + .

[0077] Compound 4: 11H NMR (600 MHz, DMSO-d6) δ 13.14 (s, 1H), 9.37 (t, J = 6.2 Hz, 1H), 8.23 - 8.22 (d, J = 1.7 Hz, 1H), 8.19 (s, 1H), 8.14 (s, 1H), 8.10 - 8.08 (m, 2H), 7.80 - 7.79 (dd, J = 8.6 Hz, 1.8 Hz, 1H), 7.75 - 7.74 (dd, J = 8.6 Hz, 1.8 Hz, 1H), 7.66 - 7.64 (d, J = 8.6 Hz, 1H), 7.27 - 7.24 (m, 1H), 6.93 - 6.92 (m, 2H), 6.84 - 6.82 (m, 1H), 4.48 - 4.47 (d, J = 6.0 Hz, 2H), 3.73 (s, 3H). 13 13C NMR (150 MHz, DMSO-d6) δ 161.60, 159.43, 140.95, 140.63, 140.10, 139.46, 138.86, 138.05, 134.10, 132.55, 129.55, 126.02, 125.79, 125.19, 123.68, 123.31, 123.07, 119.59, 118.72, 113.19, 112.31, 110.71, 55.08, 42.75. ESI-MS m / z: 414.1267 [M+H] + .

[0078] Compound 5: 1 1H NMR (600 MHz, DMSO-d6) δ 13.01 (s, 1H), 9.33 (t, J = 6.2 Hz, 1H), 8.16 - 8.14 (m, 2H), 8.10 (s, 1H), 7.99 - 7.97 (d, J = 8.2 Hz, 1H), 7.74 - 7.72 (dd, J = 8.2 Hz, 1.4 Hz, 1H), 7.63 - 7.52 (m, 1H), 7.25 (t, J = 8.3 Hz, 1H), 6.92 - 6.91 (m, 2H), 6.83 - 6.82 (m, 1H), 4.47 - 4.46 (d, J = 6.2 Hz, 2H), 3.73 (s, 3H). 13 13C NMR (150 MHz, DMSO-d6) δ 161.62, 159.43, 140.96, 140.42, 139.99, 137.85, 130.11, 129.56, 129.55, 124.83, 124.43, 123.17, 120.97, 120.82, 119.57, 119.56, 113.15, 112.31, 55.08, 42.72. ESI-MS m / z: 364.1115 [M+H]+ .

[0079] Compound 6: 1 H NMR (600MHz, DMSO-d6) δ8.92 (s, 1H), 8.74-8.73 (m, 2H), 8.36 (m, 1H), 8.17 (m, 1H), 8.15-8.13 (d, J=8.5Hz, 1H), 7.94-7.92 (m, 2H), 7.88-7.86 (m, 1H), 7.25 (t, J=8.5Hz, 1H), 6.96-6.95 (m, 2H), 6.84-6.82 (m, 1H), 4.52-4.51 (d, J=6.2Hz, 2H), 3.77 (s, 3H). 13 C NMR (150MHz, DMSO-d6) δ160.96, 159.23, 147.68, 147.35, 141.15, 141.06, 140.82, 140.24, 139.44, 133 .18, 128.71, 124.63, 124.21, 123.06, 123.02, 121.51, 121.44, 119.23, 113.11, 112.13, 54.69, 42.54. ESI-MS m / z: 375.1170[M+H] + .

[0080] Compound 7: 1 H NMR (600MHz, DMSO-d6) δ9.70 (s, 1H), 9.37 (t, J=6.2Hz, 1H), 8.56-8.55 (d, J=6.3Hz , 1H), 8.40-8.39 (d, J=8.2Hz, 1H), 8.23-8.21 (m, 2H), 8.09 (s, 1H), 8.03-8.02 (d, J= 7.2Hz, 1H), 7.97-7.94 (m, 2H), 7.60-7.59 (dd, J=8.2Hz, 1.7Hz, 1H), 7.27-7.24 (m, 1 H), 6.93-6.92 (m, 2H), 6.83-6.81 (m, 1H), 4.49-4.48 (d, J=6.0Hz, 2H), 3.74 (s, 3H). 13CNMR (150MHz, DMSO-d6) δ161.49, 159.44, 150.49, 141.20, 140.90, 140.14, 139.74, 138.57, 137.97, 134.91, 134.59, 13 4.35, 129.55, 128.99, 128.96, 128.20, 127.95, 126.34, 125.09, 123.35, 120.58, 119.59, 113.21, 112.31, 55.09, 42.77. ESI-MS m / z: 425.1332[M+H] + .

[0081] Compound 8: 1 H NMR (600MHz, DMSO-d6) δ13.14 (s, 1H), 9.37 (t, J=6.2Hz, 1H), 8.23-8.22 (d, J=1.7Hz , 1H), 8.19 (s, 1H), 8.14 (s, 1H), 8.10-8.08 (m, 2H), 7.80-7.79 (dd, J=8.6Hz, 1.8Hz, 1 H), 7.75-7.74 (dd, J=8.6Hz, 1.8Hz, 1H), 7.66-7.64 (d, J=8.6Hz, 1H), 7.27-7.24 (m, 1H), 6.93-6.92(m, 2H), 6.84-6.82(m, 1H), 4.48-4.47(d, J=6.0Hz, 2H), 3.73(s, 3H). 13 C NMR (150MHz, DMSO-d6) δ161.60, 159.43, 140.95, 140.63, 140.10, 139.46, 138.86, 138.05, 134.10, 132.55, 129 .55, 126.02, 125.79, 125.19, 123.68, 123.31, 123.07, 119.59, 118.72, 113.19, 112.31, 110.71, 55.08, 42.75. ESI-MS m / z: 414.1269[M+H] + .

[0082] Compound 9: 1H NMR (600MHz, DMSO-d6) δ13.02 (s, 1H), 9.27 (t, J = 6.2Hz, 1H), 8.32 (s, 1H), 8.27 (s, 1H), 8.09 (s, 1H), 8.05 (s, 1H), 7.90-7.89 (d, J = 8.2Hz, 1 H), 7.71-7.70 (dd, J=8.2Hz, 1.4Hz, 1H), 7.26-7.24 (m, 1H), 6.92-6.90 (m, 2H), 6.83-6.81 (m, 1H), 4.47-4.46 (d, J=6.2Hz, 2H), 3.73 (s, 3H). 13 C NMR (150MHz, DMSO-d6) δ161.67, 159.42, 141.30, 140.99, 138.87, 137.30, 136.65, 131.42, 129 .54, 126.08, 125.57, 124.92, 123.01, 120.94, 119.58, 118.33, 113.18, 112.28, 55.08, 42.72. ESI-MS m / z: 364.1117[M+H] + .

[0083] Compound 10: 1 H NMR (600MHz, DMSO-d6) δ9.43 (t, J=6.2Hz, 1H), 8.87-8.86 (d, J=6.6Hz, 2H), 8.71 (s, 1H), 8.24-8.22 (m, 3H), 8.14-8.12 (d, J=8.5Hz, 1H), 8 .14-8.12 (dd, J=8.5Hz, 1.5Hz, 1H), 7.26 (t, J=8.5Hz, 1H), 6.93-6.91 (m, 2H), 6.84-6.82 (m, 1H), 4.48-4.47 (d, J=6.2Hz, 2H), 3.73 (s, 3H). 13 C NMR (150MHz, DMSO-d6) δ161.30, 159.43, 151.69, 145.81, 145.78, 142.62, 141.13, 140.83, 140.79, 133 .26, 129.57, 126.20, 124.64, 124.18, 123.00, 122.98, 122.47, 119.63, 113.26, 112.34, 55.09, 42.81. ESI-MS m / z: 375.1165[M+H] + .

[0084] Compound 11: 11H NMR (600 MHz, DMSO-d6) δ 9.64 (s, 1H), 9.40 (t, J = 6.2 Hz, 1H), 8.56 - 8.55 (d, J = 6.3 Hz, 1H), 8.36 - 8.35 (d, J = 8.2 Hz, 1H), 8.25 (s, 1H), 8.21 (s, 1H), 8.12 - 8.11 (d, J = 8.2 Hz, 1H), 7.99 - 7.98 (dd, J = 7.2 Hz, 1.2 Hz, 1H), 7.99 - 7.98 (m, 1H), 7.93 - 7.90 (m, 2H), 7.58 - 7.56 (dd, J = 8.2 Hz, 1.7 Hz, 1H), 7.28 - 7.25 (m, 1H), 6.94 - 6.93 (m, 2H), 6.84 - 6.82 (m, 1H), 4.49 - 4.48 (d, J = 6.0 Hz, 2H), 3.74 (s, 3H). 13 13C NMR (150 MHz, DMSO-d6) δ 161.52, 159.44, 151.22, 141.00, 140.89, 140.83, 139.63, 138.96, 138.35, 135.93, 134.35, 133.59, 129.57, 129.56, 128.73, 128.48, 128.32, 127.00, 125.47, 124.71, 119.81, 119.64, 113.24, 112.34, 55.09, 42.79. ESI-MS m / z: 425.1329 [M+H] + .

[0085] Compound 12: 1 1H NMR (600 MHz, DMSO-d6) δ 13.16 (s, 1H), 9.32 (t, J = 6.2 Hz, 1H), 8.35 (s, 1H), 8.16 (s, 1H), 8.14 (s, 2H), 8.01 - 8.00 (d, J = 8.6 Hz, 1H), 7.79 - 7.76 (m, 2H), 7.65 - 7.64 (d, J = 8.6 Hz, 1H), 7.27 - 7.24 (m, 1H), 6.93 - 6.92 (m, 2H), 6.84 - 6.82 (m, 1H), 4.48 - 4.47 (d, J = 6.0 Hz, 2H), 3.74 (s, 3H). 13C NMR (150MHz, DMSO-d6) δ161.63, 159.43, 141.32, 140.96, 139.88, 139.55, 139.09, 138.01, 134.18, 132.31, 129 .55, 125.95, 125.62, 124.73, 124.48, 123.71, 120.66, 119.61, 118.92, 113.22, 112.31, 110.75, 55.09, 42.75. ESI-MS m / z: 414.1272[M+H] + .

[0086] Compound 13: 1 H NMR (600MHz, DMSO-d6) δ13.24 (s, 1H), 9.33 (t, J = 6.2Hz, 1H), 8.32 (s, 1H), 8.20 (s, 1H), 8.07 (s, 1H), 7.84-7.83 (dd, J = 7.8Hz, 1.1Hz, 1H), 7.68-7.67 (dd, J=7.8Hz, 1.1Hz, 1H), 7.47 (t, J=7.8Hz, 1H), 7.26-7.24 (m, 1H), 6.92- 6.91 (m, 2H), 6.83-6.81 (m, 1H), 4.48-4.47 (d, J=6.2Hz, 2H), 3.73 (s, 3H). 13 C NMR (150MHz, DMSO-d6) δ161.58, 159.44, 140.91, 140.16, 139.44, 137.53, 137.20, 129.55, 128 .04, 126.72, 125.70, 125.65, 124.34, 123.36, 119.60, 119.45, 113.21, 112.32, 55.09, 42.76. ESI-MS m / z: 364.1110[M+H] + .

[0087] Compound 14: 1 H NMR (600MHz, DMSO-d6) δ9.43 (t, J=6.2Hz, 1H), 8.93-8.92 (m, 2H), 8.29 (s, 1H), 8.13-8.11 (m, 3H), 7.74-7.73 (dd, J=7.4Hz, 1.1H z, 1H), 7.64 (t, J=7.4Hz, 1H), 7.26-7.23 (m, 1H), 6.91-6.90 (m, 2H), 6.83-6.81 (m, 1H), 4.47-4.46 (d, J=6.2Hz, 2H), 3.72 (s, 3H).13 C NMR (150MHz, DMSO-d6) δ161.25, 159.45, 151.14, 146.74, 146.71, 140.79, 140.77, 140.38, 138.49, 132 .06, 129.56, 127.15, 127.12, 126.18, 125.50, 124.22, 124.19, 119.61, 113.23, 112.34, 55.09, 42.79. ESI-MS m / z: 375.1177[M+H] + .

[0088] Compound 15: 1 H NMR (600MHz, DMSO-d6) 69.79 (s, 1H), 9.35 (t, J=6.2Hz, 1H), 8.52-8.51 (m, 2H), 8.2 9 (s, 1H), 8.17-8.16 (dd, J = 7.2Hz, 1.2Hz, 1H), 8.12-8.11 (dd, J = 8.2Hz, 1.1Hz, 1H) , 8.03-8.01 (m, 1H), 7.67-7.63 (m, 2H), 7.54-7.53 (dd, J=7.2Hz, 1.2Hz, 1H), 7.23- 7.20 (m, 1H), 6.87-6.86 (m, 2H), 6.81-6.79 (m, 1H), 4.43-4.41 (m, 2H), 3.71 (s, 3H). 13 CNMR (150MHz, DMSO-d6) δ161.29, 159.42, 150.68, 140.94, 140.83, 140.37, 140.00, 137.84, 136.57, 134.49, 134.28, 13 2.34, 130.13, 129.52, 129.52, 129.17, 128.18, 127.76, 125.75, 125.64, 125.54, 120.38, 119.56, 119.56, 55.07, 42.74. ESI-MS m / z: 425.1318[M+H] + .

[0089] Compound 16: 1H NMR (600MHz, DMSO-d6) δ13.26 (s, 1H), 9.32 (t, J = 6.2Hz, 1H), 8.23 ​​(s, 1H), 8.19 (s, 1H), 8.12 (s, 1H), 7.94-7.93 (dd, J = 7.6Hz, 1.6Hz, 1H), 7.72-7.68 (m, 2H), 7.57-7.52 (m, 2H), 7.26-7.23 (m, 1H), 6.91-6.90 (m, 2H), 6.82-6.81 (m, 1H), 4.46-4.45 (d, J=6.0Hz, 2H), 3.72 (s, 3H). 13 C NMR (150MHz, DMSO-d6) δ161.54, 159.42, 140.89, 140.11, 139.87, 139.66, 139.46, 136.77, 134.17, 131.87, 129 .53, 126.48, 126.22, 125.96, 125.65, 124.13, 123.29, 119.82, 119.58, 113.18, 112.30, 110.88, 55.07, 42.73. ESI-MS m / z: 414.1271[M+H] + .

[0090] Example 2

[0091] The synthetic route of benzothiophene derivatives of the present invention is as follows Figure 2 As shown, the specific preparation method is as follows:

[0092] The first step is to synthesize compound 3a-1:

[0093] Accurately weigh halogenated benzothiophene-2-carboxylic acid (i.e. Figure 2 1a-d) (1.00 g, 3.89 mmol), 6-aminohexanoic acid tert-butyl ester or 7-aminoheptanoic acid tert-butyl ester or 8-aminooctanoic acid tert-butyl ester (i.e. Figure 22a-c in (i.e., n=5, 6, 7) (0.80 g, 5.84 mmol) and N-methylimidazole (1.12 g, 13.6 mmol) were mixed and dissolved in acetonitrile (20 mL). Then, N, N, N', N'-tetramethylchloroformamidine hexafluorophosphate (1.31 g, 4.67 mmol) was added at once, and the mixture was stirred at room temperature for 24 hours. The mixture was cooled to room temperature, concentrated under reduced pressure to remove the solvent, and the residual liquid was extracted with dichloromethane, which was repeated three times and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with petroleum ether / ethyl acetate (3:1, v / v, 500 mL) to obtain compound tert-butyl halobenzothiophene carboxylate 3a-1 with yields of 80%, 85%, 85%, 80%, 80%, 80%, 85%, 85%, 80%, 80%, 85%, 80%, 80%, 85%, 80%, respectively.

[0094] The second step is to synthesize compound 4a-1:

[0095] Accurately weigh tert-butyl halobenzothiophene carboxylate (i.e. Figure 2 3a-1) (3.00 mmol) was dissolved in 10 mL of dichloromethane (CH2Cl2), and an equal volume of trifluoroacetic acid (TFA) was added. The mixture was stirred at room temperature for 6 h to obtain compound 4a-1 with a yield of 100%.

[0096] Step 3: Synthesis of compound 5a-1:

[0097] Accurately weigh tert-butyl halobenzothiophene carboxylic acid (i.e. Figure 2 4a-1) (3.00mmol), O-2-tetrahydro-2H-pyranylhydroxylamine (0.53g, 4.50mmol) and N-methylimidazole (0.86g, 10.5mmol) were mixed and dissolved in acetonitrile (20mL). Then, N, N, N', N'-tetramethylchloroformamidine hexafluorophosphate (1.01g, 3.6mmol) was added at one time, and the mixture was stirred at room temperature for 24 hours. Cooled to room temperature, concentrated under reduced pressure to remove the solvent, and the residual liquid was extracted with dichloromethane, repeated three times and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with petroleum ether / ethyl acetate (1:1, v / v, 500mL) to obtain compound 5a-1 with yields of 85%, 85%, 85%, 80%, 90%, 80%, 90%, 85%, 80%, 80%, 90%, 90%, respectively.

[0098] Step 4: Synthesis of compounds 6a-x:

[0099] A mixture of the corresponding compound 5a-1 (1.00 mmol), compound 1H-pyrazole-4-boronic acid (0.22 g, 2.00 mmol) or 4-pyridineboronic acid (0.25 g, 2.00 mmol), tetrakis(triphenylphosphine)palladium (0.11 g, 0.09 mmol) and anhydrous potassium carbonate (0.69 g, 5.00 mmol) was stirred at 100°C for 20 hours under a nitrogen atmosphere, and the solvent was a dioxane / water mixture (v / v=4:1). The mixture was cooled to room temperature, concentrated under reduced pressure to remove the solvent, and the residual liquid was extracted with dichloromethane, which was repeated three times and the organic phases were combined, dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure after filtration. The residue was purified by silica gel column chromatography with dichloromethane / methanol (40:1, v / v, 500 mL) to give compounds 6a-x with yields of 60%, 55%, 55%, 50%, 50%, 50%, 60%, 50%, 50%, 60%, 50%, 50%, 65%, 60%, 50%, 50%, 60%, 50%, 60%, 50%, 50%, 55%, 60%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%, 50%.

[0100] Step 5, synthesis of 7a-x (17-40):

[0101] Accurately weigh 6a-x (0.5 mmol) and dissolve it in 10 mL of dichloromethane. Add an equal volume of trifluoroacetic acid (TFA) and stir at room temperature for 6 h. Cool to room temperature, concentrate under reduced pressure to remove the solvent, extract the residual liquid with dichloromethane, repeat three times and combine the organic phases, dry over anhydrous sodium sulfate, filter and concentrate the filtrate under reduced pressure. The residue is purified by high performance liquid chromatography (separation conditions: preparative column Welch XB-C18, 21.2×250mm, 5μm; elution system: methanol / water (volume ratio) = 10%-100%, containing 0.5‰ trifluoroacetic acid by volume, flow rate: 10ml / min, elution time: 40min) to obtain the corresponding compounds 17-40 with a yield of 100%.

[0102] Compound 17: 1 H NMR (600MHz, DMSO-d6) δ10.33 (s, 1H), 8.87 (t, J = 5.7Hz, 1H), 8.31 (s, 1H), 8.13 (s, 2H), 7.87-7.86 (d, J = 7.6Hz, 1H), 7.51-7.50 ( dd, J=7.6Hz, 1.1Hz, 1H), 7.43 (t, J=7.6Hz, 1H), 3.27-3.24 (m, 2H), 1.95 (t, J=7.3Hz, 2H), 1.56-1.49 (m, 4H), 1.31-1.26 (m, 2H). 13C NMR (150MHz, DMSO-d6) δ169.16, 161.47, 141.23, 140.30, 136.62, 130.30, 126.41, 1 23.76, 123.45, 123.41, 120.78, 120.76, 119.62, 48.70, 32.31, 29.05, 26.24, 25.03. ESI-MS m / z: 373.1327[M+H] + .

[0103] Compound 18: 1 H NMR (600MHz, DMSO-d6) δ10.33 (s, 1H), 8.87 (t, J = 5.7Hz, 1H), 8.31 (s, 1H), 8.13 (s, 2H), 7.87-7.86 (d, J = 7.6Hz, 1H), 7.51-7.50 (d d, J=7.6Hz, J=1.1Hz, 1H), 7.43 (t, J=7.6Hz, 1H), 3.27-3.24 (m, 2H), 1.93 (t, J=7.3Hz, 2H), 1.55-1.46 (m, 4H), 1.33-1.25 (m, 4H). 13 C NMR (150MHz, DMSO-d6) δ169.18, 161.45, 141.20, 140.28, 136.60, 130.28, 126.39, 123. 73, 123.43, 123.43, 120.76, 120.74, 119.60, 48.69, 32.32, 29.16, 28.44, 26.34, 25.16. ESI-MS m / z: 387.1484[M+H] + .

[0104] Compound 19: 1 H NMR (600MHz, DMSO-d6) 610.33 (s, 1H), 8.87 (t, J = 5.7Hz, 1H), 8.32 (s, 1H), 8.14 (s, 2H), 7.87-7.86 (d, J = 7.6Hz, 1H), 7.51-7.50 (d d, J=7.6Hz, J=1.1Hz, 1H), 7.43 (t, J=7.6Hz, 1H), 3.27-3.24 (m, 2H), 1.92 (t, J=7.3Hz, 2H), 1.53-1.45 (m, 4H), 1.29-1.21 (m, 6H). 13C NMR (150MHz, DMSO-d6) δ169.26, 161.48, 141.24, 140.33, 136.64, 130.28, 126.41, 123.77, 123.45, 123.44, 120.78, 120.73, 119.65, 48.71, 32.36, 29.27, 28.66, 28.61, 26.55, 25.21. ESI-MS m / z: 401.1651[M+H] + .

[0105] Compound 20: 1 H NMR (600MHz, DMSO-d6) δ10.33 (s, 1H), 8.92-8.91 (m, 2H), 8.80 (t, J=5.7Hz, 1H), 8.19-8.17 (d, J=7.8Hz, 1H), 8.16 (s, 1H ), 7.99-7.97(m, 2H), 7.62-7.57(m, 2H), 3.25-3.22(m, 2H), 1.93(t, J=7.3Hz, 2H), 1.52-1.47(m, 4H), 1.29-1.23(m, 2H). 13 CNMR (150MHz, DMSO-d6) δ169.16, 161.16, 158.69, 158.46, 151.64, 146.33, 142.16, 141.43, 1 36.79, 134.41, 126.41, 125.99, 125.50, 124.50, 122.36, 48.70, 32.29, 28.97, 26.20, 24.98. ESI-MS m / z: 384.1387[M+H] + .

[0106] Compound 21: 1 H NMR (600MHz, DMSO-d6) δ10.33 (s, 1H), 8.92-8.90 (m, 2H), 8.80 (t, J=5.7Hz, 1H), 8.19-8.18 (d, J=7.8Hz, 1H), 8.16 (s, 1H ), 7.97-7.96 (m, 2H), 7.63-7.57 (m, 2H), 3.25-3.22 (m, 2H), 1.92 (t, J=7.3Hz, 2H), 1.51-1.44 (m, 4H), 1.30-1.22 (m, 4H). 13CNMR (150MHz, DMSO-d6) δ169.17, 161.13, 158.53, 158.29, 151.31, 146.60, 142.11, 141.39, 136. 78, 134.48, 126.38, 125.93, 125.37, 124.41, 122.35, 48.68, 32.29, 29.07, 28.38, 26.28, 25.12. ESI-MS m / z: 398.1536[M+H] + .

[0107] Compound 22: 1 H NMR (600MHz, DMSO-d6) δ10.33 (s, 1H), 8.93-8.92 (m, 2H), 8.81 (t, J=5.7Hz, 1H), 8.19-8.18 (d, J=7.8Hz, 1H), 8.16 (s, 1H ), 7.99-7.98(m, 2H), 7.63-7.57(m, 2H), 3.25-3.22(m, 2H), 1.91(t, J=7.3Hz, 2H), 1.51-1.43(m, 4H), 1.27-1.18(m, 6H). 13 CNMR (150MHz, DMSO-d6) δ169.22, 161.14, 158.65, 158.41, 151.57, 146.39, 142.16, 141.42, 136.79, 134.42, 126.39, 125.97, 125.47, 124.47, 122.34, 48.69, 32.33, 29.17, 28.61, 28.54, 26.48, 25.16. ESI-MS m / z: 412.1699[M+H] + .

[0108] Compound 23: 1 H NMR (600MHz, DMSO-d6) δ12.99 (s, 1H), 10.33 (s, 1H), 8.75 (t, J=5.7Hz, 1H), 8.66 (s, 1H), 8.11 (d, J=1.6Hz, 1H), 8.01 (s, 2H), 7.97-7.95 (d, J=8.2Hz, 1H), 7.72-7.70 (dd, J=8.2Hz, 1.6Hz, 1H), 3.26-3.23 (m, 2H), 1.95 (t, J=7.4Hz, 2H), 1.55-1.49 (m, 4H), 1.33-1.26 (m, 2H). 13C NMR (150MHz, DMSO-d6) δ169.14, 161.41, 140.86, 139.99, 137.73, 130.05, 124.37, 1 24.28, 123.16, 123.14, 120.98, 120.71, 120.71, 39.23, 32.30, 28.92, 26.18, 24.97. ESI-MS m / z: 373.1335[M+H] + .

[0109] Compound 24: 1 H NMR (600MHz, DMSO-d6) δ12.97 (s, 1H), 10.33 (s, 1H), 8.74 (t, J=5.7Hz, 1H), 8.66 (s, 1H), 8.11 (d, J=1.6Hz, 1H), 8.01 (s, 2H), 7.97-7 .95 (d, J=8.2Hz, 1H), 7.72-7.70 (dd, J=8.2Hz, 1H), 3.26-3.23 (m, 2H), 1.93 (t, J=7.4Hz, 2H), 1.54-1.46 (m, 4H), 1.33-1.25 (m, 4H). 13 C NMR (150MHz, DMSO-d6) δ169.19, 161.42, 140.87, 140.00, 137.73, 130.05, 124.38, 124. 28, 123.15, 123.14, 121.00, 120.73, 120.72, 39.18, 32.32, 29.06, 28.42, 26.30, 25.17. ESI-MS m / z: 387.1493[M+H] + .

[0110] Compound 25: 1 H NMR (600MHz, DMSO-d6) δ12.97 (s, 1H), 10.32 (s, 1H), 8.75 (t, J=5.7Hz, 1H), 8.65 (s, 1H), 8.12-8.11 (d, J=1.6Hz, 1H), 8.01 (s, 2H), 7.97-7 .95 (d, J=8.2Hz, 1H), 7.72-7.70 (dd, J=8.2Hz, 1.6Hz, 1H), 3.27-3.23 (m, 2H), 1.92 (t, J=7.4Hz, 2H), 1.53-1.45 (m, 4H), 1.30-1.22 (m, 6H). 13C NMR (150MHz, DMSO-d6) δ169.20, 161.41, 140.88, 139.99, 137.72, 130.05, 124.36, 124.27, 123.14, 123.14, 120.99, 120.72, 120.72, 39.13, 32.34, 29.14, 28.64, 28.56, 26.47, 25.16. ESI-MS m / z: 401.1640[M+H] + .

[0111] Compound 26: 1 H NMR (600MHz, DMSO-d6) δ10.35 (s, 1H), 8.90-8.87 (m, 3H), 8.54 (d, J = 1.6Hz, 1H), 8.27-8.26 (m, 2H), 8.24-8.22 (d, J = 8.2Hz, 1H), 8 .17 (s, 1H), 8.01-7.99 (dd, J=8.3Hz, 1.6Hz, 1H), 3.28-3.25 (m, 2H), 1.95 (t, J=7.3Hz, 2H), 1.56-1.50 (m, 4H), 1.32-1.27 (m, 2H). 13 C NMR (150MHz, DMSO-d6) δ169.15, 161.13, 158.53, 158.30, 145.33, 142.39, 142.19, 140.11, 1 32.30, 125.00, 124.75, 124.56, 124.08, 123.14, 123.12, 48.69, 32.30, 28.90, 26.19, 24.98. ESI-MS m / z: 384.1380[M+H] + .

[0112] Compound 27: 1 H NMR (600MHz, DMSO-d6) δ10.33 (s, 1H), 8.88-8.85 (m, 3H), 8.53-8.52 (d, J=1.6Hz, 1H), 8.24-8.22 (d, J=8.2Hz, 1H), 8.22-8.21 (m, 2H ), 8.17 (s, 1H), 8.00-7.98 (dd, J=8.3Hz, 1.6Hz, 1H), 3.28-3.25 (m, 2H), 1.94 (t, J=7.3Hz, 2H), 1.56-1.47 (m, 4H), 1.33-1.22 (m, 4H). 13C NMR (150MHz, DMSO-d6) 6169.18, 161.13, 158.31, 158.09, 145.96, 142.21, 142.13, 140.10, 132. 51, 124.97, 124.73, 124.43, 124.04, 122.94, 122.92, 48.68, 32.31, 29.01, 28.41, 26.30, 25.16. ESI-MS m / z: 398.1548[M+H] + .

[0113] Compound 28: 1 H NMR (600MHz, DMSO-d6) δ10.34 (s, 1H), 8.91-8.89 (m, 3H), 8.56 (d, J = 1.6Hz, 1H), 8.32-8.31 (m, 2H), 8.24-8.23 (d, J = 8.2Hz, 1H), 8 .18 (s, 1H), 8.02-8.00 (dd, J=8.3Hz, 1.6Hz, 1H), 3.28-3.25 (m, 2H), 1.93 (t, J=7.3Hz, 2H), 1.54-1.45 (m, 4H), 1.30-1.23 (m, 6H). 13 C NMR (150MHz, DMSO-d6) δ169.22, 161.11, 158.59, 158.36, 144.79, 142.54, 142.26, 140.12, 132.09, 125.01, 124.74, 124.69, 124.11, 123.32, 123.32, 48.69, 32.34, 29.10, 28.65, 28.56, 26.48, 25.17. ESI-MS m / z: 412.1695[M+H] + .

[0114] Compound 29: 1 H NMR (600MHz, DMSO-d6) 610.34 (s, 1H), 8.69 (t, J = 5.7Hz, 1H), 8.24 (s, 1H), 8.17 (s, 2H), 8.00 (s, 1H), 7.87 (d, J = 8.30H z, 1H), 7.70 (dd, J=8.30, 1.50H, 1H), 3.26-3.22 (m, 2H), 1.95 (t, J=7.3Hz, 2H), 1.54-1.49 (m, 4H), 1.31-1.26 (m, 2H). 13C NMR (150MHz, DMSO-d6) δ169.15, 161.49, 141.18, 139.33, 137.31, 131.28, 125.47, 1 25.46, 124.46, 122.96, 120.95, 118.33, 118.32, 48.69, 32.31, 28.94, 26.18, 24.98. ESI-MS m / z: 373.1324[M+H] + .

[0115] Compound 30: 1 H NMR (600MHz, DMSO-d6) 610.32 (s, 1H), 8.67 (t, J = 5.5Hz, 1H), 8.24 (s, 1H), 8.17 (s, 2H), 8.00 (s, 1H), 7.87 (d, J = 8.5 Hz, 1H), 7.70 (dd, J=8.5, 1.6H, 1H), 3.26-3.22 (m, 2H), 1.93 (t, J=7.3Hz, 2H), 1.54-1.46 (m, 4H), 1.33-1.25 (m, 4H). 13 C NMR (150MHz, DMSO-d6) δ169.18, 161.48, 141.17, 139.33, 137.30, 131.27, 125.45, 125. 44, 124.44, 122.94, 120.94, 118.31, 118.31, 48.67, 32.31, 29.06, 28.41, 26.28, 25.16. ESI-MSm / z: 387.1487[M+H] + .

[0116] Compound 31: 1 H NMR (600MHz, DMSO-d6) δ13.00 (s, 1H), 10.31 (s, 1H), 8.67 (t, J=5.7Hz, 1H), 8.64 (s, 1H), 8.30 (s, 1H), 8.24 (s, 1H), 8.04 (s, 1H), 8.00 (s, 1 H), 7.88 (d, J=8.30Hz, 1H), 7.70 (dd, J=8.30, 1.30H, 1H), 3.26-3.23 (m, 2H), 1.93 (t, J=7.3Hz, 2H), 1.54-1.45 (m, 4H), 1.30-1.22 (m, 6H). 13C NMR (150MHz, DMSO-d6) δ169.20, 161.48, 141.17, 139.34, 137.31, 131.27, 125.46, 125.45, 124.44, 122.95, 120.95, 118.32, 118.32, 48.69, 32.34, 29.16, 28.65, 28.57, 26.47, 25.18. ESI-MS m / z: 401.1642[M+H] + .

[0117] Compound 32: 1 H NMR (600MHz, DMSO-d6) 610.35 (s, 1H), 8.86-8.84 (m, 1H), 8.68 (s, 1H), 8.20-8.19 (d, J=6.6Hz, 2H), 8.14-8.11 (m, 2H), 7.98 (dd, J=8.5Hz, 1.7H, 1H), 3.28-3.24 (m, 2H), 1.95 (t, J=7.3Hz, 2H), 1.56-1.49 (m, 4H), 1.32-1.27 (m, 2H). 13 C NMR (150MHz, DMSO-d6) δ169.60, 161.59, 158.77, 158.54, 146.56, 143.44, 141.49, 141.24, 1 33.71, 126.54, 124.64, 124.57, 123.34, 123.33, 122.82, 49.13, 32.74, 29.31, 26.62, 25.41. ESI-MS m / z: 384.1374[M+H] + .

[0118] Compound 33: 1 H NMR (600MHz, DMSO-d6) δ10.33 (s, 1H), 8.84-8.83 (m, 3H), 8.68 (s, 1H), 8.19-8.18 (m, 2H), 8.13 (s, 1H), 8.11 (d, J=8.3Hz , 1H), 7.98 (dd, J=8.3Hz, J=1.5Hz, 1H), 3.28-3.25 (m, 2H), 1.94 (t, J=7.3Hz, 2H), 1.55-1.47 (m, 4H), 1.34-1.25 (m, 4H). 13CNMR (150MHz, DMSO-d6) δ169.16, 161.13, 158.25, 158.02, 146.29, 142.97, 141.03, 140.76, 133. 32, 126.07, 124.18, 124.10, 122.83, 122.83, 122.34, 48.67, 32.31, 28.99, 28.41, 26.28, 25.15. ESI-MS m / z: 398.1539[M+H] + .

[0119] Compound 34: 1 H NMR (600MHz, DMSO-d6) δ10.33 (s, 1H), 8.83 (t, J=5.63Hz, 1H), 8.80-8.79 (m, 2H), 8.63 (s, 1H), 8.13 (s, 1H), 8.10-8.09 (m, 3H), 7.95 (dd, J=8.5Hz, 1.5H, 1H), 3.28-3.25 (m, 2H), 1.93 (t, J=7.3Hz, 2H), 1.55-1.45 (m, 4H), 1.30-1.23 (m, 6H). 13 C NMR (150MHz, DMSO-d6) δ169.21, 161.17, 158.39, 158.17, 147.17, 142.77, 141.05, 140.58, 133.68, 126.03, 124.19, 124.04, 122.57, 122.55, 122.12, 48.69, 32.34, 29.09, 28.65, 28.57, 26.48, 25.18. ESI-MS m / z: 412.1701[M+H] + .

[0120] Compound 35: 1 H NMR (600MHz, DMSO-d6) 610.33 (s, 1H), 8.74 (t, J=5.7Hz, 1H), 8.18 (s, 2H), 8.11 (s, 1H), 7.83-7.81 (m, 1H), 7.67-7 .65 (m, 1H), 7.46 (t, J=7.8Hz, 1H), 3.27-3.24 (m, 2H), 1.95 (t, J=7.4Hz, 2H), 1.55-1.49 (m, 4H), 1.32-1.22 (m, 2H). 13C NMR (150MHz, DMSO-d6) δ169.14, 161.38, 140.15, 139.85, 137.40, 127.99, 125.64, 1 25.63, 125.17, 124.19, 123.23, 123.22, 119.44, 48.68, 32.29, 28.89, 26.16, 24.97. ESI-MS m / z: 373.1333[M+H] + .

[0121] Compound 36: 1 H NMR (600MHz, DMSO-d6) 610.33 (s, 1H), 8.74 (t, J=5.7Hz, 1H), 8.18 (s, 2H), 8.11 (s, 1H), 7.83-7.81 (m, 1H), 7.66-7 .65 (m, 1H), 7.46 (t, J=7.8Hz, 1H), 3.27-3.23 (m, 2H), 1.93 (t, J=7.4Hz, 2H), 1.55-1.47 (m, 4H), 1.33-1.23 (m, 4H). 13 C NMR (150MHz, DMSO-d6) δ169.19, 161.39, 140.17, 139.88, 137.41, 127.99, 125.63, 125. 18, 125.18, 124.20, 123.26, 123.24, 119.46, 48.69, 32.32, 29.04, 28.42, 26.29, 25.17. ESI-MS m / z: 387.1490[M+H] + .

[0122] Compound 37: 1 H NMR (600MHz, DMSO-d6) 610.33 (s, 1H), 8.76 (t, J=5.7Hz, 1H), 8.19 (s, 2H), 8.11 (s, 1H), 7.83-7.81 (m, 1H), 7.67-7 .65 (m, 1H), 7.46 (t, J=7.8Hz, 1H), 3.27-3.24 (m, 2H), 1.92 (t, J=7.4Hz, 2H), 1.54-1.45 (m, 4H), 1.29-1.23 (m, 6H). 13C NMR (150MHz, DMSO-d6) δ169.21, 161.39, 140.18, 139.90, 137.41, 128.00, 125.64, 125.60, 125.17, 124.21, 123.26, 123.26, 119.47, 48.70, 32.35, 29.14, 28.66, 28.58, 26.48, 25.19. ESI-MSm / z: 401.1648[M+H] + .

[0123] Compound 38: 1 H NMR (600MHz, DMSO-d6) δ10.34 (s, 1H), 8.90-8.89 (m, 2H), 8.83 (t, J=5.7Hz, 1H), 8.20 (s, 1H), 8.11-8.09 (m, 1H), 8.07-8.06 (m, 2H), 7.72-7.71 (m, 1H), 7.64 (t, J=7.9Hz, 1H), 3.27-3.23 (m, 2H), 1.94 (t, J=7.4Hz, 2H), 1.55-1.49 (m, 4H), 1.31-1.26 (m, 2H). 13 C NMR (150MHz, DMSO-d6) δ169.58, 161.49, 158.91, 158.67, 150.95, 147.74, 141.21, 138.82, 1 32.68, 127.37, 127.30, 126.57, 125.49, 124.46, 124.43, 49.13, 32.74, 29.31, 26.60, 25.41. ESI-MS m / z: 384.1365[M+H] + .

[0124] Compound 39: 1 H NMR (600MHz, DMSO-d6) δ10.35 (s, 1H), 8.96-8.95 (m, 2H), 8.85 (t, J = 5.7Hz, 1H), 8.21 (s, 1H), 8.18-8.17 (m, 2H), 8.11 (d, J = 7.9Hz, 1H), 7.74 (d, J=7.9Hz, 1H), 7.64 (t, J=7.9Hz, 1H), 3.27-3.23 (m, 2H), 1.94 (t, J=7.4Hz, 2H), 1.54-1.46 (m, 4H), 1.30-1.25 (m, 4H). 13C NMR (150MHz, DMSO-d6) δ169.30, 161.08, 158.83, 158.59, 151.97, 146.04, 140.88, 138.41, 131. 83, 127.21, 127.17, 126.16, 125.07, 124.52, 124.51, 48.71, 32.35, 29.02, 28.45, 26.31, 25.21. ESI-MS m / z: 398.1533[M+H] + .

[0125] Compound 40: 1 H NMR (600MHz, DMSO-d6) δ10.31 (s, 1H), 8.89-8.88 (m, 2H), 8.82 (t, J=5.7Hz, 1H), 8.18 (s, 1H), 8.09-8.08 (m, 1H), 8.08-8.07 (m, 2H), 7.70-7.69 (m, 1H), 7.61 (t, J=7.9Hz, 1H), 3.24-3.21 (m, 2H), 1.89 (t, J=7.4Hz, 2H), 1.51-1.42 (m, 4H), 1.26-1.18 (m, 6H). 13 C NMR (150MHz, DMSO-d6) 6169.20, 161.03, 158.53, 158.29, 150.80, 147.03, 140.80, 138.37, 132.14, 126.95, 126.92, 126.11, 125.02, 124.08, 124.03, 48.68, 32.33, 29.07, 28.64, 28.54, 26.44, 25.16. ESI-MS m / z: 412.1686[M+H] + .

[0126] Test Example 1: Breast Cancer Inhibitory Activity

[0127] MAD-MB-231 breast cancer cells in logarithmic growth phase were collected and cultured at 8 × 10 3The density of cells / well was seeded in a 96-well plate. After overnight culture, drugs were added. SAHA (Vorinostat) and Y-27632 were used as positive drugs. Three sub-wells were set for each concentration and culture was continued for 72 hours. After 72 hours, the 96-well plate was taken out, and 100 μL of 10% trichloroacetic acid aqueous solution was added to each well after discarding the culture medium, and fixed in a 4°C refrigerator for at least 2 hours. After fixation, the 96-well plate was taken out, the trichloroacetic acid solution was discarded, and it was rinsed at least 4 times with slowly flowing water and dried. Then 80 μL of 0.4% SRB solution prepared with 1% acetic acid was added and stained for 20 minutes. During this period, 1% acetic acid aqueous solution was prepared. After staining, it was rinsed 4 times with 1% acetic acid aqueous solution to remove unbound dyes and dried again. After the 96-well plate was completely dried, 100 μL of 10mM Tris solution was added to each well, and the dye bound to the protein was completely dissolved by shaking, and the absorbance was detected at 515nm in an ELISA reader. Inhibition rate = (OD value of control group - OD value of drug-treated group) / OD value of control group × 100%, Graphicpad Prism 9 software was used to calculate the IC value of the inhibitory activity of the compound. 50 See Table 1 for values.

[0128] Table 1 Inhibitory activity of the compounds prepared in the present invention on MDA-MB-231 breast cancer cells

[0129] Compound <![CDATA[IC 50 (μM)]]> Compound <![CDATA[IC 50 (μM)]]> 1 ＞10.00 21 2.90±0.30 2 ＞10.00 22 ＞10.00 3 ＞10.00 23 ＞10.00 4 ＞10.00 24 0.37±0.01 5 ＞10.00 25 6.42±1.10 6 ＞10.00 26 ＞10.00 7 ＞10.00 27 0.13±0.02 8 ＞10.00 28 0.42±0.03 9 ＞10.00 29 ＞10.00 10 ＞10.00 30 1.26±0.01 11 ＞10.00 31 0.76±0.01 12 ＞10.00 32 ＞10.00 13 ＞10.00 33 0.95±0.09 14 ＞10.00 34 0.46±0.03 15 ＞10.00 35 ＞10.00 16 ＞10.00 36 9.053±1.20 17 3.21±0.09 37 ＞10.00 18 ＞10.00 38 ＞10.00 19 ＞10.00 39 1.54±0.01 20 ＞10.00 40 ＞10.00 Y-27632 ＞10.00 SAHA 0.66±0.02

[0130] Conclusion: Among the disclosed compounds, 24, 27, 28, and 34 have better anti-proliferation activity against MDA-MB-231 breast cancer cells than the positive drugs SAHA and Y-27632.

[0131] Detection Example 2 ROCK activity test

[0132] The following method is used to determine the inhibitory effect of the compounds prepared by the present invention on the activity of ROCK2 kinase. The experimental method is briefly described as follows: The ROCK2 kinase inhibitory activity is determined by using the HTRF KinEASE kit (PerkinElmer, catalog number: 62ST2PEB) based on time-resolved fluorescence technology. Y-27632 and SAHA are used as positive controls, and three replicate wells are set up in parallel. 4 μL of the test compound or buffer, 2 μL of the reaction substrate S2, 2 μL of ROCK2 kinase and 2 μL of ATP are added to each well of the white 384-well plate in sequence. Incubate in a 37°C incubator for 30 minutes. 5 μL of streptokinase-labeled XL-665 and 5 μL of EuK-labeled anti-phosphorylated protein kinase antibody are added in sequence, reacted at room temperature for 60 minutes, and the fluorescence signal is detected by a Tecan multifunctional microplate reader. The fluorescence intensity value of the sample well measured at λ=665nm is F1, and the fluorescence intensity value at λ=620nm is F2. The signal ratio calculation formula is: signal ratio=F1 / F2×10 4The inhibitory rate of the test compound on protein kinase is calculated as follows: Inhibition rate (%) = [1-(test compound signal ratio-minimum signal ratio) / (maximum signal ratio-minimum signal ratio)] × 100. The maximum signal ratio is the solvent control, and the minimum signal ratio is the signal ratio of the XL-665 well without streptokinase labeling. The inhibitory effect of ROCK1 kinase activity, the experimental method is the same as ROCK2 activity test, but the incubation time is changed to 20min. The IC of the inhibitory activity of the compound was calculated using Graghpadprism 9 software 50 See Table 2 for values.

[0133] Table 2 Inhibitory activity of the compounds prepared by the present invention on ROCK kinase

[0134] Compound <![CDATA[ROCK2 IC 50 (nM)]]> <![CDATA[ROCK1 IC 50 (nM)]]> 1 ＞1000 ＞1000 2 ＞1000 ＞1000 3 ＞1000 ＞1000 4 ＞1000 ＞1000 5 6.751±0.3 15.79±5.5 6 ＞1000 ＞1000 7 ＞1000 ＞1000 8 ＞1000 ＞1000 9 7.935±2.0 64.26±11.6 10 295.9±19.8 565.5±30.7 12 ＞1000 ＞1000 13 ＞1000 ＞1000 14 224.7±8.6 2035±447.5 15 ＞1000 ＞1000 16 ＞1000 ＞1000 24 254.9±21.7 58.18±20.9 27 ＞1000 ＞1000 28 ＞1000 ＞1000 34 ＞1000 ＞1000 SAHA ＞1000 ＞1000 Y-27632 409.2±26.2 32.86±3.6

[0135] Conclusion: Among the compounds prepared by the present invention, 5, 9, 10, 14, and 24 have better ROCK2 kinase inhibitory activity than the positive drug Y27632; among the compounds prepared by the present invention, 5 has better ROCK1 kinase inhibitory activity than the positive drug Y27632; among the compounds 24, 27, 28, and 34 that have better anti-MDA-MB-231 breast cancer cell proliferation activity than the positive drugs SAHA and Y27632, compound 24 exhibits efficient ROCK1 and 2 kinase inhibitory activity. Test Example 3 Evaluation of HDAC Inhibitory Activity

[0136] In the experimental group, 5 μL of different concentrations of compound 24 (1 μM, 333.3 nM, 111.1 nM, 37.0 nM, 12.3 nM, 4.1 nM, 1.4 nM, 0.5 nM, 0.2 nM) and HDAC enzyme diluent were added to a black 96-well plate and incubated at 37°C for 5 min. In the blank group, 30 μL of solvent was added to the wells. Then, 20 μL of fluorescent substrate (Ac-Leu-Gly-Lys(Ac)-AMC for HDAC1-3, 6 and 11; Ac-Leu-Gly-Lys(tfa)-AMC for HDAC4-5 and 7-9; Ac-Arg-His-Lys(Ac)-Lys(Ac)-AMC for HDAC10) was added to all wells and incubated at 37°C for 60 min. Then, 50 μL of pancreatin solution was added to each well, and the fluorescence value was measured under a microplate reader after incubation for 30 min. Each group was measured in parallel three times. The IC value of the inhibitory activity of the compound was calculated using Graphicpad Prism 9 software. 50 See Table 3 for values.

[0137] Table 3 Inhibitory activity of compound 24 and SAHA on HDAC1-11

[0138] Compound 24 SAHA <![CDATA[HDAC1 IC 50 (nM)]]> 9.09±0.32 85.13±8.11 <![CDATA[HDAC2 IC 50 (nM)]]> 8.03±2.61 47.88±1.91 <![CDATA[HDAC3 IC 50 (nM)]]> 6.26±3.38 83.48±6.95 <![CDATA[HDAC4 IC 50 (nM)]]> ＞10000.00 ＞10000.00 <![CDATA[HDAC5 IC 50 (nM)]]> ＞10000.00 ＞10000.00 <![CDATA[HDAC6 IC 50 (nM)]]> 0.41±0.02 13.61±1.86 <![CDATA[HDAC7 IC 50 (nM)]]> ＞10000.00 ＞10000.00 <![CDATA[HDAC8 IC 50 (nM)]]> 7.69±2.33 480.80±28.61 <![CDATA[HDAC9 IC 50 (nM)]]> ＞10000.00 ＞10000.00 <![CDATA[HDAC10 IC 50 (nM)]]> ＞10000.00 ＞10000.00 <![CDATA[HDAC11 IC 50 (nM)]]> ＞10000.00 ＞10000.00

[0139] Conclusion: Compound 24 prepared by the present invention has better HDAC1, 2, 3, 6, 8 inhibitory activity than the positive drug SAHA. Compound 24 not only has efficient HDAC1, 2, 3, 6, 8 and ROCK1, 2 inhibitory activity, but also has better anti-MDA-MB-231 breast cancer cell proliferation activity than the positive drugs SAHA and Y27632. Test Example 4 Evaluation of T cell activity in tumor tissue

[0140] 4T1 cells (1×10 per mouse 5 cells) were injected orthotopically into the mammary glands of Balb / c mice. When the tumor volume reached 50 mm 3 The mice were randomly divided into four groups. After oral administration of compound 24 (50 mg / kg), compound 24 (25 mg / kg), SAHA (50 mg / kg) and saline control group for 15 consecutive days, the mice were killed, and the tumor tissues of the mice in each group were collected and soaked in PBS. Grind, collect through a 40μm cell sieve, centrifuge and adjust the cell suspension to an appropriate concentration with PBS for use. After washing with PBS twice, resuspend the cells with 200μL PBS. When the cells are in a suspension state, put them into the flow tube, label the flow tube, add Anti-Mouse CD45 MAB PerCP / Cy5.5, APC Anti-Mouse CD3 Antibody[17A2], FITC Anti-Mouse CD4Antibody[GK1.5], PE Anti-Mouse CD8a Antibody to the labeled flow tubes, mix well, and incubate at room temperature in the dark for 30 minutes. Then add 500μL PBS to wash twice to remove the unlabeled antibodies, and finally resuspend the cells with 400μL PBS. Transfer the cell suspension to the dedicated tube of the flow cytometer for detection. The detection results are as follows: Figure 3 shown.

[0141] Conclusion: Compared with the control group, the CD3 + T cells, CD4 + T cells and CD8 + Compared with the SAHA (50 mg / kg) treatment group, the CD3 + T cells, CD4 + T cells and CD8 + The proportion of T cells increased by 1.36 times, 1.82 times, and 1.21 times, respectively. Compound 24 has a better anti-tumor immune effect than the positive drug SAHA.

[0142] The compound 24 prepared by the present invention not only has highly effective HDAC1, 2, 3, 6, 8 and ROCK1, 2 inhibitory activity, but also has better anti-MDA-MB-231 breast cancer cell proliferation activity and in vivo anti-tumor immune activity than positive drugs.

[0143] Finally, it should be noted that the terms "comprises," "includes," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements that are inherent to such process, method, article, or apparatus.

[0144] Although the preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0145] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A benzothiophene derivative, characterized in that: The structure of the benzothiophene derivative is shown in Formula I: wherein the R1 group is at least one of a hydrogen atom, a halogen group, a pyrazolyl group, a pyridyl group, an isoquinolyl group and an indolyl group, The R2 group is at least one of the following groups: hydrogen atom, halogen group, benzyl group, 5-7 membered aryl group, heteroaryl group, cycloalkyl group, heterocyclic group, alkyl group with 1 to 6 carbon atoms, haloalkyl group, haloalkoxy group, alkylhydroxamic acid, hydroxyalkyl group, cyano group, hydroxyl group, nitro group, carboxyl group.

2. The benzothiophene derivative according to claim 1, characterized in that: The R1 group is pyrazolyl, pyridyl, isoquinolyl or indolyl, The R2 group is a 5- to 7-membered aryl group or an alkyl group having 1 to 6 carbon atoms.

3. The benzothiophene derivative according to claim 2, characterized in that: The benzothiophene derivative is any one of the following compounds:

4. The method for preparing the benzothiophene derivatives according to any one of claims 1 to 3, characterized in that: The following steps are involved: (1) A halogenated benzothiophene-2-carboxylic acid and an amine having a corresponding R2 group are subjected to an amidation reaction under the action of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate to obtain product 1; (2) product 1 undergoes a Suzuki reaction with a boric acid having a corresponding R1 group to obtain the benzothiophene derivative; The R1 group is pyrazolyl, pyridyl, isoquinolyl or indolyl, The R2 group is a 5- to 7-membered aryl group.

5. The method for preparing benzothiophene derivatives according to claim 4, characterized in that: In step (1), the molar ratio of the halogenated benzothiophene-2-carboxylic acid to the amine having the corresponding R2 group is 1:1.5; During the reaction, a catalyst is also added, and the catalyst is N-methylimidazole; In step (2), product 1 and a boric acid having a corresponding R1 group are dissolved in a solvent, and then a catalyst and a base are added to react. The molar ratio of the product 1 to the boronic acid having the corresponding R1 group is 1:2; The solvent is a mixed solution of dioxane and water, The catalyst is tetrakis(triphenylphosphine)palladium, The base is potassium carbonate.

6. The method for preparing the benzothiophene derivatives according to any one of claims 1 to 3, characterized in that: The following steps are involved: (1) subjecting a halogenated benzothiophene-2-carboxylic acid and an ester having a corresponding R2 group to an amidation reaction under the action of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate to obtain product 1; (2) subjecting product 1 to a deprotection reaction to obtain product 2; (3) subjecting product 2 to an amidation reaction with O-2-tetrahydro-2H-pyranylhydroxylamine in the presence of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate to obtain product 3; (4) Product 3 undergoes Suzuki reaction with a boronic acid having a corresponding R1 group to obtain product 4; (5) subjecting the product 4 to a deprotection reaction to obtain the benzothiophene derivative; The R1 group is pyrazolyl or pyridyl, The R2 group is an alkyl group having 1 to 6 carbon atoms.

7. The method for preparing benzothiophene derivatives according to claim 6, characterized in that: In step (1), the molar ratio of the halogenated benzothiophene-2-carboxylic acid to the ester having the corresponding R2 group is 1:1.5; During the reaction, a catalyst is also added, and the catalyst is N-methylimidazole; In steps (2) and (5), the deprotection reaction is carried out by dissolving product 1 or product 4 in dichloromethane and adding trifluoroacetic acid in an equal volume to dichloromethane; In step (3), the molar ratio of product 2 to O-2-tetrahydro-2H-pyranolamine is 1:1.5; In step (4), the product 3 and the boric acid having the corresponding R1 group are dissolved in a solvent, and then a catalyst and a base are added to react. The molar ratio of the product 3 to the boronic acid having the corresponding R1 group is 1:2; The solvent is a mixed solution of dioxane and water, The catalyst is tetrakis(triphenylphosphine)palladium, The base is potassium carbonate.

8. Use of the benzothiophene derivative according to any one of claims 1 to 3 in the preparation of a drug for treating and / or preventing diseases associated with abnormal activity of ROCK and / or HDAC.

9. The use according to claim 8, characterized in that: The disease associated with abnormal activity of ROCK and / or HDAC is selected from at least one of cancer, inflammatory diseases, autoimmune diseases, fibrotic diseases, transplant rejection, diseases associated with excessive secretion of IL-17, IL-21 and / or IL-23, eye diseases, cardiovascular and cerebrovascular diseases, central nervous system diseases, diabetic nephropathy and skin diseases.

10. The use according to claim 9, characterized in that: The disease associated with abnormal activity of ROCK and / or HDAC is breast cancer, endometrial cancer, ovarian cancer, prostate cancer, colorectal cancer, lung cancer, rheumatoid arthritis or psoriasis.

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