A benzothiophene derivative, its preparation method and application

Abstract

This invention discloses a benzothiophene derivative, its preparation method, and its applications, relating to the field of pharmaceutical technology. This invention provides a novel class of benzothiophene derivatives with ROCK kinase inhibitory activity and dual ROCK / HDAC inhibitory activity. Experimental results demonstrate that the compounds provided by this invention exhibit high activity, and the prepared compounds possess good ROCK / HDAC inhibitory activity, particularly showing good activity in inhibiting breast cancer cell proliferation and in the immune response. Furthermore, this invention proposes a method for preparing this benzothiophene derivative, which features a short synthetic route, a simple and easy-to-operate preparation process, and can meet the needs of large-scale industrial production.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a benzothiophene derivative, its preparation method, and its application. Background Technology

[0002] Rho-associated coiled-coil protein 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 C-terminal PH domain bisected by a zinc finger-like motif domain (CRD). In the classic ROCK signaling pathway, Rho GTPases activate downstream ROCK kinases, which further phosphorylate downstream ROCK substrates, thereby remodeling the cytoskeleton, inducing actin-myosin contraction, and regulating microtubule dynamics.

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

[0004] ROCK kinases are involved in the development and progression of various diseases, including eye diseases, cardiovascular and cerebrovascular diseases, tumor metastases, central nervous system diseases, and diabetic nephropathy, making them important targets for the treatment of many diseases. To date, four ROCK inhibitors have successfully completed clinical trials and been marketed: fasudil, ribasudil, netarsudil, and KD025. In addition, the ROCK inhibitor Y27632 is currently undergoing a Phase I clinical trial for the treatment of corneal edema, while AT13148 has completed a Phase I study for the treatment of advanced solid tumors. However, due to the narrow therapeutic index and pharmacokinetic characteristics 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 containing multiple subclasses and members, playing crucial roles in various cellular biological processes. HDACs function in cell cycle, cell differentiation, apoptosis, DNA repair, and cell metabolism, among others. They influence cell fate and function by regulating the expression of specific genes. Based on sequence homology, they are classified into four classes: Class I HDACs (HDAC1, HDAC2, HDAC3, HDAC8), Class IIa HDACs (HDAC4, HDAC5, HDAC7, HDAC9), Class IIb HDACs (HDAC6, HDAC10), Class III HDACs (Sirtuins), and Class IV HDACs (HDAC11).

[0006] To date, numerous HDAC inhibitors have been developed, six of which have been approved for clinical use: Voronostat (SAHA), Belinostat, Panobinostat, Pracinostat, Chidamide, and Romidepsin. However, the use of HDAC inhibitors as single-agent treatments is limited. Past research has often combined HDAC inhibitors with other protein inhibitors to form dual-target inhibitors, thereby enhancing therapeutic efficacy.

[0007] The interaction between ROCK and HDAC in various pathological processes offers novel therapeutic strategies for treating a wide range of diseases. These enzymes play crucial roles in cancer, inflammation, autoimmune diseases, fibrosis, transplant rejection, diseases associated with specific cytokines such as IL-17, IL-21, and IL-23, as well as ocular, cardiovascular, central nervous system, diabetic nephropathy, and skin diseases. They collectively influence disease development and progression by regulating cellular behavior, inflammatory responses, and immune responses. Dual-target inhibitors, as an innovative therapeutic strategy, offer a more effective treatment approach by simultaneously acting on both ROCK and HDAC, two key enzymes. This approach targets multiple stages of disease development, thereby improving efficacy while reducing the side effects and drug resistance risks that may arise from single-target therapy. Summary of the Invention

[0008] Based on the shortcomings of the existing technology, the applicant has designed a benzothiophene derivative or its pharmaceutical salt with ROCK kinase inhibitory activity and a ROCK / HDAC dual-target inhibitory activity, as well as the preparation method and application of the benzothiophene derivative or its pharmaceutical salt.

[0009] This 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 the following: a hydrogen atom substituted at one position on the benzene ring, a halogen group, a pyrazolyl group, a pyridyl group, an isoquinolinyl group, and an indole group.

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

[0013] Preferably, the R1 group is a pyrazolyl, pyridyl, isoquinolinyl, or indole group substituted at one of the positions on the benzene ring (positions 4-7 of the benzothiophene ring).

[0014] The R2 group is an aryl group with 5 to 7 members or an alkyl group with 1 to 6 carbon atoms.

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

[0016]

[0017]

[0018]

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

[0020]

[0021] Experimental results in specific embodiments of the present invention show that compounds 24, 27, 28, and 34 exhibit better anti-MDA-MB-231 breast cancer cell proliferation activity than the positive control drugs SAHA and Y27632. Compound 24 also demonstrates more efficient ROCK1 and 2 kinase inhibitory activity. Furthermore, compound 24 exhibits better anti-tumor immunomodulatory effects than the positive control drug SAHA.

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

[0023] (1) The product 1 was prepared by amidation of halobenzothiophene-2-carboxylic acid and amine with corresponding R2 group under the action of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate.

[0024] (2) The product 1 reacts with boric acid having the corresponding R1 group in a Suzuki reaction (also known as Suzuki coupling reaction, Suzuki-Miyaura reaction) to prepare the benzothiophene derivative;

[0025] The R1 group is a pyrazolyl, pyridyl, isoquinolinyl, or indole group 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 halobenzothiophene-2-carboxylic acid and amine having the corresponding R2 group is 1:1.5;

[0028] During the reaction, a catalyst, namely N-methylimidazole, was also added.

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

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

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

[0032] The catalyst is tetra(triphenylphosphine)palladium.

[0033] The alkali is potassium carbonate.

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

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

[0036] (1) The product 1 was prepared by amidation of esters containing halobenzothiophene-2-carboxylic acid and corresponding R2 groups under the action of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate.

[0037] (2) Deprotection reaction of product 1 to obtain product 2;

[0038] (3) Product 2 was prepared by amidation reaction of product 2 with O-2-tetrahydro-2H-pyranolamine in the presence of N,N,N′,N′-tetramethylchloromethamine hexafluorophosphate;

[0039] (4) Product 3 reacts with boric acid having the corresponding R1 group in a Suzuki reaction to obtain product 4;

[0040] (5) Deprotect product 4 to obtain the benzothiophene derivative;

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

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

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

[0044] During the reaction, a catalyst, namely N-methylimidazole, was also added.

[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-pyranolamine is 1:1.5;

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

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

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

[0050] The catalyst is tetra(triphenylphosphine)palladium.

[0051] The alkali is potassium carbonate.

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

[0053] The present invention also provides the use of the benzothiophene derivatives in the preparation of medicaments for the treatment and / or prevention of diseases associated with abnormal ROCK and / or HDAC activity.

[0054] Specifically, the diseases associated with abnormal ROCK and / or HDAC activity are selected from at least one of the following: 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 ROCK and / or HDAC activity include breast cancer, endometrial cancer, ovarian cancer, prostate cancer, colorectal cancer, lung cancer, rheumatoid arthritis, or psoriasis. In cancer, inhibiting ROCK and / or HDAC can simultaneously suppress cancer cell migration, invasion, proliferation, and angiogenesis, thereby effectively inhibiting tumor growth and spread. In inflammatory diseases, it can reduce inflammatory responses by inhibiting the migration of inflammatory cells and the expression of inflammatory genes. In autoimmune diseases, it can reduce the immune system's attack on its own tissues by regulating the activity of immune cells and the expression of immune-related genes. Although the examples only demonstrate the therapeutic effect on breast cancer, based on the mechanisms of action of ROCK and HDAC in various diseases, it is speculated that this benzothiophene derivative can also be used to treat other diseases associated with abnormal ROCK and / or HDAC activity, such as endometrial cancer, ovarian cancer, prostate cancer, colorectal cancer, lung cancer, rheumatoid arthritis, and psoriasis.

[0056] Experimental results in specific embodiments of the present invention show that compounds 24, 27, 28, and 34 exhibit better anti-MDA-MB-231 breast cancer cell proliferation activity than the positive control drugs SAHA and Y27632. Compound 24 also demonstrates more efficient ROCK1 and 2 kinase inhibitory activity. Furthermore, compound 24 exhibits better anti-tumor immunomodulatory effects than the positive control drug SAHA.

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

[0058] The active compounds can be formulated into forms suitable for administration via any suitable route, using one or more pharmaceutically acceptable carriers through conventional methods. Therefore, the active compounds of the present invention can be formulated into various dosage forms for oral administration, injection (e.g., intravenous, intramuscular, or subcutaneous), inhalation, or insufflation. The compounds 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 guideline, the active compound is preferably administered in unit dose form or by self-administration as a single dose by the patient. The unit dose of the compound or composition of the present invention may be expressed as a tablet, capsule, sachet, bottled liquid, powder, granule, lozenge, suppository, regenerated powder, or liquid formulation. A suitable unit dose may be 0.1–1000 mg.

[0060] In addition to the active compound, the pharmaceutical composition of the present invention may contain one or more excipients, wherein the excipients are selected from the following components: fillers (diluents), binders, wetting agents, disintegrants, or excipients, etc. Depending on the method of administration, 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 activity of the specific compound used, the severity of the disease, the patient's age, the patient's weight, the patient's health status, the patient's behavior, the patient's diet, the timing of administration, the route of administration, the rate of excretion, and the combination of drugs; in addition, the optimal treatment mode, such as the treatment pattern, daily dosage, or the type of medicinal salt, can be validated based on conventional treatment protocols.

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

[0063] This invention provides a novel class of benzothiophene derivatives with ROCK kinase inhibitory activity and dual ROCK / HDAC inhibitory activity. Experimental results demonstrate that the compounds provided by this invention exhibit high activity, and the prepared compounds possess excellent ROCK / HDAC inhibitory activity, particularly showing good activity against breast cancer cell proliferation and in the immune response. Furthermore, this invention also proposes a method for preparing these benzothiophene derivatives. This method features a short synthetic route, a simple and easy-to-operate process, and can meet the needs of large-scale industrial production. Attached Figure Description

[0064] Figure 1 This is a synthetic route diagram for the A-type benzothiophene derivatives of the present invention.

[0065] Figure 2 This is a synthetic route diagram for the B-type benzothiophene derivatives of the present invention.

[0066] Figure 3 The effects of compounds 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 Implementation

[0067] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Unless otherwise specified, the experimental methods described in the following embodiments are conventional methods; the reagents and biological materials described are commercially available unless otherwise specified.

[0068] Example 1

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

[0070] Step 1, synthesize 3a-d:

[0071] Accurately weigh the halobenzothiophene-2-carboxylic acid (i.e. Figure 1 The 1a-d (1.00g, 3.89mmol) and m-methylbenzylamine (i.e. Figure 1 Compound 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′-tetramethylchloromethanesulfonyl hexafluorophosphate (1.31 g, 4.67 mmol) was added in a single batch, and the mixture was stirred at room temperature for 24 hours. After cooling to room temperature, the solvent was removed by concentration under reduced pressure, and the residue was extracted with dichloromethane. The extraction was repeated three times and the organic phases were combined. The residue was 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 give compounds 3a-d in yields of 80%, 85%, 75%, and 80%, respectively.

[0072] The second step is to synthesize 4a-p(1-16):

[0073] The corresponding compounds 3a-d (0.50 mmol) and compound 1H-pyrazole-4-boronic acid (i.e. Figure 1 X1 (0.11 g, 1.00 mmol) or 4-pyridineboronic acid (i.e. Figure 1 X2 (0.12g, 1.00mmol) or isoquinoline-5-boronic acid (i.e. Figure 1 X3 (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 with a dioxane / water mixture (v / v = 4:1). After cooling to room temperature, the solvent was removed by concentration under reduced pressure. The residue was extracted with dichloromethane, repeated three times, and the organic phases were combined. The residue was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was then separated by high-performance liquid chromatography (HPLC) (separation conditions: preparative column, Welch). XB-C18, 21.2×250mm, 5μm; elution system: methanol / water (volume ratio) = 10%-100%, containing 0.5‰ trifluoroacetic acid (volume percentage), flow rate: 10ml / min, elution time: 40min) purification, yielding the corresponding compounds 1-16 in 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 the benzothiophene derivatives of the present invention is as follows: Figure 2 As shown, the specific preparation method is as follows:

[0092] Step 1: Synthesize compound 3a-1:

[0093] Accurately weigh the halobenzothiophene-2-carboxylic acid (i.e. Figure 2 The 1a-d (1.00g, 3.89mmol) in the formula, tert-butyl 6-aminohexanoate or tert-butyl 7-aminoheptanoate or tert-butyl 8-aminooctanoate (i.e. Figure 22a-c (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′-tetramethylchloroformamidin hexafluorophosphate (1.31 g, 4.67 mmol) was added in one batch, and the mixture was stirred at room temperature for 24 hours. After cooling to room temperature, the solvent was removed by concentration under reduced pressure, and the residue was extracted with dichloromethane. The extraction was repeated three times and the organic phases were combined. The residue was 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 give compound tert-butyl halobenzothiophene carboxylate 3a-1 in yields of 80%, 85%, 85%, 80%, 80%, 80%, 85%, 85%, 80%, 80%, 85%, 80%, 85%, 80%.

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

[0095] Accurately weigh tert-butyl halobenzothiophene carboxylate (i.e. Figure 2 3a-l (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 give compound 4a-l, with a yield of 100%.

[0096] The third step is to synthesize compound 5a-1:

[0097] Accurately weigh tert-butyl halobenzothiophene carboxylic acid (i.e. Figure 2 4a-l (3.00 mmol), O-2-tetrahydro-2H-pyranolamine (0.53 g, 4.50 mmol), and N-methylimidazole (0.86 g, 10.5 mmol) were mixed and dissolved in acetonitrile (20 mL). Then, N,N,N′,N′-tetramethylchloromethanesulfonyl hexafluorophosphate (1.01 g, 3.6 mmol) was added in a single batch, and the mixture was stirred at room temperature for 24 hours. After cooling to room temperature, the solvent was removed by concentration under reduced pressure, and the residue was extracted with dichloromethane. The extraction was repeated three times and the organic phases were combined. The residue was 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 give compound 5a-l in yields of 85%, 85%, 85%, 80%, 90%, 80%, 90%, 85%, 80%, 80%, 90%, 90%.

[0098] Step 4: Synthesize compound 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 h under a nitrogen atmosphere with a dioxane / water mixture (v / v = 4:1). After cooling to room temperature, the solvent was removed by concentration under reduced pressure. The residual liquid was extracted with dichloromethane, and the mixture was repeated three times. 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 in dichloromethane / methanol (40:1, v / v, 500 mL) to give compounds 6a-x in yields of 60%, 55%, 55%, 50%, 55%, 60%, 50%, 50%, 60%, 60%, 50%, 50%, 65%, 60%, 50%, 50%, 60%, 60%, 50%, 50%, 55%, 60%, 50%, 50%.

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

[0101] Accurately weigh 0.5 mmol of 6a-x 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 residue with dichloromethane, repeat three times and combine the organic phases, dry to anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure. Separate the residue 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 (volume percentage), flow rate: 10ml / min, elution time: 40min) purification, to obtain the corresponding compounds 17-40, yield 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] Example 1: Breast Cancer Suppressive Activity

[0127] Log-growth phase MAD-MB-231 breast cancer cells were collected and stored at 8 × 10⁻⁶. 3Cells were seeded at a density of 1:1 in 96-well plates and cultured overnight. Afterward, positive control agents were added, with SAHA (Vorinostat) and Y-27632 used as agents. Three auxiliary wells were prepared for each concentration, and the plates were cultured for another 72 hours. After 72 hours, the 96-well plates were removed, the culture medium was discarded, and 100 μL of 10% trichloroacetic acid aqueous solution was added to each well. The plates were fixed at 4°C for at least 2 hours. After fixation, the 96-well plates were removed, the trichloroacetic acid solution was discarded, and the plates were rinsed at least four times with slowly flowing water and dried. Then, 80 μL of 0.4% SRB solution prepared with 1% acetic acid was added for staining for 20 minutes. During staining, a 1% acetic acid aqueous solution was prepared. After staining, the plates were rinsed four times with 1% acetic acid aqueous solution to remove unbound dye and dried again. Once the 96-well plates were completely dry, 100 μL of 10 mM Tris solution was added to each well, and the plates were shaken to completely dissolve the dye bound to the proteins. The absorbance was measured at 515 nm using a microplate reader. Inhibition rate = (OD value of control group - OD value of treated group) / OD value of control group × 100%. The IC50 of the compound's inhibitory activity is calculated using Graghpad Prism 9 software. 50 The values ​​are shown in Table 1.

[0128] Table 1. Inhibitory activity of the compounds prepared in this invention against 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: Compounds 24, 27, 28, and 34 in this study exhibit better anti-MDA-MB-231 breast cancer cell proliferation activity than the positive control drugs SAHA and Y-27632.

[0131] Example 2: ROCK activity test

[0132] The following method was used to determine the inhibitory effect of the compound prepared in this invention on ROCK2 kinase activity. The experimental method is briefly described below: ROCK2 kinase inhibitory activity was measured using the HTRF KinEASE kit (PerkinElmer, catalog number: 62ST2PEB) based on time-resolved fluorescence technology. Y-27632 and SAHA were used as positive controls, with three replicates. 4 μL of the test compound or buffer, 2 μL of reaction substrate S2, 2 μL of ROCK2 kinase, and 2 μL of ATP were added sequentially to each well of a white 384-well plate. The plate was incubated at 37°C for 30 min. Then, 5 μL of streptokinase-labeled XL-665 and 5 μL of EuK-labeled antiphosphorylated protein kinase antibody were added sequentially, and the reaction was carried out at room temperature for 60 min. The fluorescence signal was detected using a Tecan multi-mode microplate reader. The fluorescence intensity value measured at λ = 665 nm was F1, and the fluorescence intensity value at λ = 620 nm was F2. The signal ratio was calculated using the formula: Signal ratio = F1 / F2 × 10⁻⁶. 4The inhibition rate of the test compound against protein kinase was calculated using the formula: Inhibition rate (%) = [1 - (signal ratio of test compound - minimum signal ratio) / (maximum signal ratio - minimum signal ratio)] × 100. Where the maximum signal ratio is the solvent control, and the minimum signal ratio is the signal ratio of the XL-665 wells without streptokinase labeling. The inhibitory effect on ROCK1 kinase activity was tested using the same method as the ROCK2 activity assay, but the incubation time was changed to 20 min. The IC50 of the inhibitory activity was calculated using Graghpadprism 9 software. 50 The values ​​are shown in Table 2.

[0133] Table 2. Inhibitory activity of the compounds prepared in this invention against 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] Conclusions: Compounds 5, 9, 10, 14, and 24 prepared in this invention exhibit better ROCK2 kinase inhibitory activity than the positive control drug Y27632; compound 5 prepared in this invention exhibits better ROCK1 kinase inhibitory activity than the positive control drug Y27632; among compounds 24, 27, 28, and 34, which show better anti-MDA-MB-231 breast cancer cell proliferation activity than the positive control drugs SAHA and Y27632, compound 24 exhibits highly efficient ROCK1 and ROCK2 kinase inhibitory activity. 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 dilution buffer were added to black 96-well plates 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 (HDAC1-3, 6 and 11 are Ac-Leu-Gly-Lys(Ac)-AMC; HDAC4-5 and 7-9 are Ac-Leu-Gly-Lys(tfa)-AMC; HDAC10 is Ac-Arg-His-Lys(Ac)-Lys(Ac)-AMC) was added to all wells and incubated at 37 °C for 60 min. Subsequently, 50 μL of trypsin solution was added to each well, and incubation was continued for 30 min. The fluorescence value was then measured using a microplate reader. Each group was measured in triplicate. The IC50 of the inhibitory activity of the compound was calculated using Graghpad Prism 9 software. 50 The values ​​are shown in Table 3.

[0137] Table 3. Inhibitory activity of compound 24 and SAHA against 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 in this invention exhibits better HDAC1, 2, 3, 6, and 8 inhibitory activity than the positive control drug SAHA. Compound 24 not only possesses highly efficient inhibitory activity against HDAC1, 2, 3, 6, and 8 and ROCK1, 2, but also shows better anti-MDA-MB-231 breast cancer cell proliferation activity than the positive control drugs SAHA and Y27632. Example 4: Evaluation of T cell activity in tumor tissue.

[0140] 4T1 cells (1×10⁴ cells per mouse) 5 (One cell) was injected orally into the mammary glands of Balb / c mice. When the tumor volume reached 50 mm... 3 Mice were randomly divided into four groups. Mice were administered compound 24 (50 mg / kg), compound 24 (25 mg / kg), SAHA (50 mg / kg), and a saline control group orally for 15 consecutive days. Mice were then sacrificed, and tumor tissues from each group were collected and soaked in PBS. Cells were ground, passed through a 40 μm cell sieve, centrifuged, and adjusted to an appropriate concentration with PBS for later use. Cells were washed twice with PBS and resuspended in 200 μL of PBS. When the cells were in suspension, they were transferred to flow cytometry tubes, which were labeled. Anti-Mouse CD45 MAB PerCP / Cy5.5, APC Anti-Mouse CD3 Antibody [17A2], FITC Anti-Mouse CD4 Antibody [GK1.5], and PE Anti-Mouse CD8a Antibody were added to the labeled flow cytometry tubes, mixed thoroughly, and incubated at room temperature in the dark for 30 min. Then, wash twice with 500 μL PBS to remove unlabeled antibodies. Finally, resuspend the cells in 400 μL PBS, transfer the cell suspension to flow cytometry tubes, and perform analysis. The results are shown below. Figure 3 As shown.

[0141] Conclusion: Compared with the control group, CD3 levels were significantly lower in the 24 (50 mg / kg), 24 (25 mg / kg), and SAHA (50 mg / kg) treatment groups. + T cells, CD4 + T cells and CD8 + The proportion of T cells increased significantly. Compared with the SAHA (50 mg / kg) treatment group, the CD3 count in the 24 (50 mg / kg) treatment group was significantly higher. + 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, and compound 24 had a better anti-tumor immune effect than the positive control drug SAHA.

[0142] Compound 24 prepared in this invention not only has highly efficient inhibitory activities against HDAC1, 2, 3, 6, 8 and ROCK1, 2, but also has better anti-MDA-MB-231 breast cancer cell proliferation activity and in vivo anti-tumor immune activity than positive control drugs.

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

[0144] Although preferred embodiments of the invention have been described, those skilled in the art, upon learning the basic inventive concept, can make other changes and modifications to these embodiments. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments as well as all changes and modifications falling within the scope of the invention.

[0145] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A benzothiophene derivative, characterized in that, The structures of the benzothiophene derivatives are shown in Formula I: Formula I, Wherein, the R1 group is pyrazolyl, pyridyl, isoquinolinyl, or indoleyl. The R2 group is benzyl.

2. A benzothiophene derivative, characterized in that, The benzothiophene derivative is any one of the following compounds: 。 3. The method for preparing the benzothiophene derivative according to claim 1, characterized in that, Includes the following steps: (1) Product 1 was prepared by amidation of halogenated benzothiophene-2-carboxylic acid and amine with corresponding R2 groups in the presence of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate. (2) Product 1 reacts with boric acid having the corresponding R1 group in a Suzuki reaction to obtain the benzothiophene derivative; The R1 group is pyrazolyl, pyridyl, isoquinolinyl, or indoleyl, and the R2 group is benzyl.

4. The method for preparing benzothiophene derivatives according to claim 3, characterized in that, In step (1), the molar ratio of halobenzothiophene-2-carboxylic acid and amine having the corresponding R2 group is 1:1.5; During the reaction, a catalyst, namely N-methylimidazole, was also added. In step (2), product 1 and boric acid with the corresponding R1 group are dissolved in a solvent, and then a catalyst and a base are added to carry out the reaction. The molar ratio of product 1 to boric acid having the corresponding R1 group is 1:2; The solvent is a mixed solution of dioxane and water. The catalyst is tetra(triphenylphosphine)palladium. The alkali is potassium carbonate.

5. A method for preparing a benzothiophene derivative, characterized in that, The preparation method is as follows: (1) The first step is an amidation reaction in the presence of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate; (2) The second step involves a deprotection reaction; (3) The third step involves an amidation reaction in the presence of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate; (4) The Suzuki reaction occurs in the fourth step; (5) In the fifth step, a deprotection reaction occurs to obtain the benzothiophene derivative.

6. The method for preparing the benzothiophene derivative according to claim 5, characterized in that, In step (1), the molar ratio of halobenzothiophene-2-carboxylic acid and the ester with the corresponding substituent group is 1:1.5; During the reaction, a catalyst, namely N-methylimidazole, was also added. In steps (2) and (5), the deprotection reaction is carried out by dissolving 3a-l or 6a-x in dichloromethane and adding trifluoroacetic acid in an equal volume to dichloromethane; In step (3), the molar ratio of 4a-l to O-2-tetrahydro-2H-pyranolamine is 1:1.5; In step (4), 5a-l and boric acid with the corresponding substituent groups are dissolved in a solvent, and then a catalyst and a base are added to carry out the reaction. The molar ratio of 5a-l to boric acid with the corresponding substituent group is 1:2; The solvent is a mixed solution of dioxane and water. The catalyst is tetra(triphenylphosphine)palladium. The alkali is potassium carbonate.

7. The use of the benzothiophene derivative of claim 1 or 2 in the preparation of a medicament for treating and / or preventing diseases associated with abnormal ROCK and / or HDAC activity, wherein the diseases associated with abnormal ROCK and / or HDAC activity are 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.

8. The application according to claim 7, characterized in that, The diseases associated with abnormal ROCK and / or HDAC activity are breast cancer, endometrial cancer, ovarian cancer, prostate cancer, colorectal cancer, lung cancer, rheumatoid arthritis, or psoriasis.

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