Benzothiophene ROCK2 inhibitor as well as preparation method and application thereof

Abstract

The invention discloses a benzothiophene ROCK2 inhibitor as well as a preparation method and application thereof. The invention provides a benzothiophene derivative which is novel in structure and has ROCK2 kinase inhibition activity. Tests prove that the compounds provided by the invention all show relatively high activity, the prepared compounds have good ROCK2 inhibitory activity, and particularly, ROCK2 / ROCK1 selectivity is higher than that of a positive drug KD025. The ROCK2 inhibitor is a benzothiophene derivative, can be used for treating diseases or symptoms related to ROCK2 activity, and can be used for preparing anti-tumor / tumor immunity enhancing and / or anti-inflammatory drugs for treating and preventing tumors and / or inflammations. 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] This invention relates to the field of pharmaceutical technology, specifically to a benzothiophene derivative that can be used as a ROCK2 inhibitor, 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, such as eye diseases, cardiovascular and cerebrovascular diseases, tumor metastasis, central nervous system diseases, and diabetic nephropathy. They are important targets for the treatment of many diseases. For example, patent CN118055926A discloses two compounds that are ROCK2 inhibitors. These compounds or pharmaceutical compositions can be used to treat or prevent ROCK2-related diseases and conditions (e.g., fibrotic diseases, autoimmune diseases, inflammatory fibrosis, inflammatory diseases, edema, eye diseases, cardiovascular diseases, central nervous system diseases, and cancer). RhoGTPase-mediated signaling pathways play a crucial role in balancing and coordinating T cell-mediated immune processes, including T cell development, activation, and differentiation. Recent studies have shown that overactivation of ROCK kinases also plays an indispensable role in inflammatory and autoimmune diseases, such as psoriasis, inflammatory bowel disease, rheumatoid arthritis, and systemic lupus erythematosus. For instance, patent CN107019698A discloses the application of ROCK2 inhibitors in the preparation of drugs for inflammatory bowel disease. Dysregulated ROCK kinases primarily affect autoimmune processes by modulating the dynamic cytoskeleton, with ROCK2 modulating autoimmunity by controlling the balance between pro-inflammatory and anti-inflammatory T cell populations.

[0005] To date, four ROCK inhibitors have successfully completed clinical trials and entered the market: 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. Summary of the Invention

[0006] In a first aspect, the present invention provides a benzothiophene-type ROCK2 inhibitor, which is a compound having the chemical structure shown in Formula I and / or a pharmaceutically acceptable salt thereof:

[0007]

[0008] In Formula I, R1 is at least one of hydrogen atom, halogen group, pyrazolyl group, and pyridinyl group, and R2 is at least one of methoxyphenyl, trifluoromethoxy, difluoromethoxy, phenethoxy, 4-chloro-1-methoxyphenyl, 4-fluoro-1-methoxyphenyl, and 4-(trifluoromethyl)-1-methoxyphenyl.

[0009] And / or, compounds having the chemical structure shown in Formula II and / or their pharmaceutically acceptable salts:

[0010]

[0011] In Formula II, R3 is at least one of hydrogen atom, halogen group, and pyrazol group, and n is 2 or 3.

[0012] The benzothiophene ROCK2 inhibitors may be selected from at least one of the following compounds or their pharmaceutically acceptable salts:

[0013]

[0014]

[0015]

[0016]

[0017] Preferably, the present invention selects compounds 5, 8, 37, 38, 39, and 40; data from the embodiments of the present invention show that compounds 5, 8, 37, 38, 39, and 40 have good inhibitory activity against ROCK2 kinase, and the ROCK2 / ROCK1 selectivity is stronger than that of the positive control drug KD025.

[0018] In a second aspect, the present invention provides the use of the benzothiophene ROCK2 inhibitor described in the first aspect in the preparation of a drug for inhibiting ROCK2.

[0019] Thirdly, the present invention provides the use of the benzothiophene ROCK2 inhibitors described in the first aspect in the preparation of medicaments for treating and / or preventing diseases associated with abnormal ROCK2 activity.

[0020] The disease described may be inflammation and / or cancer.

[0021] As a key member of the Rho-associated kinase family, ROCK2 significantly enhances cell contractility and promotes actin cytoskeleton remodeling by phosphorylating downstream substrates (such as MLC and LIMK), thereby driving the abnormal migration and invasion of tumor cells. Furthermore, dysregulation of the ROCK2 signaling pathway disrupts cell polarity regulation, exacerbates the formation of an inflammatory microenvironment, and indirectly supports tumor proliferation and metastasis by inhibiting apoptosis and promoting epithelial-mesenchymal transition (EMT). It is noteworthy that these mechanisms are widely associated in a variety of malignant tumors, including breast cancer, endometrial cancer, ovarian cancer, vaginal cancer, fallopian tube cancer, cervical cancer, kidney cancer, bladder cancer, urothelial carcinoma, urethral cancer, prostate cancer, testicular cancer, colorectal cancer, sarcoma, bone cancer, multiple myeloma, leukemia, myxoma, rhabdomyosarcoma, leiomyosarcoma, fibroma, lipoma, teratoma, pharyngeal cancer, nasopharyngeal carcinoma, oral cancer, lung cancer, alveolar carcinoma, lymphoma, mesothelioma, small intestine cancer, gastric cancer, esophageal cancer, pancreatic cancer, liver cancer, bile duct cancer, neurofibroma, glioma, neuroblastoma, neuroblastoma, melanoma, skin cancer, basal cell carcinoma, squamous cell carcinoma, thyroid cancer, head and neck cancer, salivary gland cancer, and at least one of gastrointestinal stromal tumors.

[0022] ROCK2 promotes the production and release of various pro-inflammatory factors (such as TNF-α, IL-6 and IL-1β) by regulating the activity of inflammatory signaling pathways (such as NF-κB and STAT3). At the same time, it can also enhance the permeability of vascular endothelial cells and promote the migration and infiltration of inflammatory cells, thereby amplifying the inflammatory response in local tissues and disrupting immune homeostasis. It is noteworthy that these mechanisms are widely associated with a variety of inflammatory diseases, including at least one of atopic dermatitis, enteritis, psoriasis, psoriatic arthritis, encephalitis, keratitis, conjunctivitis, rhinitis, otitis media, gingivitis, pharyngitis, tonsillitis, pneumonia, hepatitis, dysentery, prostatitis, endometritis, cervicitis, pelvic inflammatory disease, paronychia, myocarditis, rheumatoid arthritis, osteoarthritis, juvenile idiopathic arthritis, ankylosing spondylitis, allergic airway disease, chronic obstructive pulmonary disease, asthma, bronchitis, systemic lupus erythematosus, cutaneous lupus erythematosus, lupus nephritis, dermatomyositis, autoimmune liver disease, Sjögren's syndrome, multiple sclerosis, dry eye disease, diabetes and related complications, thyroiditis, contact dermatitis, and amyotrophic lateral sclerosis.

[0023] In the embodiments of the present invention, compound 40 not only exhibits good activity and selectivity, but also demonstrates better efficacy in treating atopic dermatitis than the positive control drug KD025 and abuxitinib at the same dose.

[0024] Fourthly, the present invention provides a pharmaceutical composition comprising one or more of a pharmaceutically acceptable carrier, a diluent, and an excipient, as well as the benzothiophene ROCK2 inhibitor described in the first aspect.

[0025] Fifthly, the present invention provides a method for preparing the benzothiophene-based ROCK2 inhibitor described in the first aspect.

[0026] The benzothiophene-based ROCK2 inhibitor is a compound having the chemical structure shown in Formula I, and its synthetic route A includes:

[0027] In synthetic route A, an amidation reaction is carried out between a halobenzothiophene-2-carboxylic acid and an amine having the corresponding M2 group in the presence of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate to obtain product 3a-1; product 3a-1 undergoes a Suzuki reaction (also known as the Suzuki-Miyaura reaction) with boric acid having the corresponding M3 group to obtain a compound with the chemical structure shown in Formula I; the M2 group is methoxyphenyl, and the M3 group is pyrazolyl or pyridinyl; or, its synthetic route B includes:

[0028] In synthetic route B, an amidation reaction is carried out between a halobenzothiophene-2-carboxylic acid and an amine having the corresponding M5 group in the presence of N,N,N′,N′-tetramethylchloromethamidine hexafluorophosphate to obtain product 6a-1; product 6a-1 undergoes a Suzuki reaction (also known as the Suzuki-Miyaura reaction) with the corresponding pyrazolium boric acid to give a compound having the chemical structure shown in Formula I; the M5 group is at least one of trifluoromethoxy, difluoromethoxy, phenethoxy, 4-chloro-1-methoxyphenyl, 4-fluoro-1-methoxyphenyl, and 4-(trifluoromethyl)-1-methoxyphenyl; or,

[0029] The benzothiophene-based ROCK2 inhibitor is a compound having the chemical structure shown in the formula, and its synthetic route C includes:

[0030] In synthetic route C, halogenated benzothiophene-2-carboxylic acid and 3-(aminomethyl)phenol undergo an amidation reaction in the presence of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate to yield products 8a-b. Products 8a-b then undergo nucleophilic substitution with 2-bromo-N,N-dimethylethylamine or 3-bromo-N,N-dimethylpropylamine in the presence of Cs₂CO₃ to generate products 9a-d. Products 9a-d then undergo the Suzuki reaction (also known as the Suzuki-Miyaura reaction) with the corresponding pyrazolboronic acid to give compounds with the chemical structures shown in Formula II.

[0031] Specifically, in synthetic route A, the molar ratio of halobenzothiophene-2-carboxylic acid and amine having the corresponding M2 group is 1:1.5; a first catalyst is also added during the reaction; product 3a-1 and boric acid having the corresponding M3 group are dissolved in a solvent, and then a second catalyst and a base are added to carry out the reaction, wherein the molar ratio of product 3a-1 and boric acid having the corresponding M3 group is 1:2;

[0032] In synthetic route B, the molar ratio of halobenzothiophene-2-carboxylic acid and amine having the corresponding M5 group is 1:1.5; a first catalyst is also added during the reaction; product 6a-1 and pyrazoleboronic acid are dissolved in a solvent, and then a second catalyst and a base are added to react, wherein the molar ratio of product 6a-1 and pyrazoleboronic acid is 1:2.

[0033] In synthetic route C, the molar ratio of halobenzothiophene-2-carboxylic acid and 3-(aminomethyl)phenol is 1:1.5; a first catalyst is also added during the reaction; the molar ratio of product 8a-b to 2-bromo-N,N-dimethylethylamine or 3-bromo-N,N-dimethylpropylamine is 1:2; product 9a-d and pyrazolonic acid are dissolved in a solvent, and then a second catalyst and a base are added to react, wherein the molar ratio of product 9a-d to pyrazolonic acid is 1:2.

[0034] The first catalyst is N-methylimidazole, the second catalyst is tetra(triphenylphosphine)palladium, the solvent is a mixed solution of dioxane and water, and the base is potassium carbonate.

[0035] Compared with the prior art, the beneficial effects of this invention are as follows: the benzothiophene ROCK2 inhibitor of this invention has strong ROCK2 inhibitory activity, and this type of compound has the advantages of readily available reaction raw materials and easy preparation. In the field of preparing antitumor or anti-inflammatory drugs, it can be used as an antitumor or anti-inflammatory therapeutic agent. Attached Figure Description

[0036] Figure 1 This is a synthetic route diagram for the A-type benzothiophene derivatives of the present invention.

[0037] Figure 2 This is a synthetic route diagram for the B-type benzothiophene derivatives of the present invention.

[0038] Figure 3 This is a C-synthetic route diagram for the benzothiophene derivatives of the present invention.

[0039] Figure 4 The figure shows the efficacy test results of the drug in treating atopic dermatitis in female Balb / c mice. Detailed Implementation

[0040] 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.

[0041] Example 1

[0042] The synthetic route for benzothiophene derivatives in this embodiment is as follows: Figure 1 As shown, the specific preparation method is as follows:

[0043] Step 1, Synthesis of 3a-1:

[0044] Accurately weigh the halobenzothiophene-2-carboxylic acid (i.e. Figure 1 1a-d (1.00 g, 3.89 mmol), amines with the corresponding M2 group (i.e. Figure 1Compound 2a-c (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 compound 3a-1 in yields of 86%, 86%, 85%, 76%, 86%, 86%, 80%, 78%, 88%, 86%, 85%, and 76%, respectively.

[0045] The second step is to synthesize 4a-x(1-24):

[0046] A mixture of the corresponding compound 3a-1 (0.50 mmol), compound 1H-pyrazole-4-boronic acid (X1) (0.11 g, 1.00 mmol) or 4-pyridineboronic acid (X2) (0.12 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 subjected to 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 1-24 in yields of 76%, 82%, 83%, 76%, 82%, 86%, 81%, 79%, 78%, 82%, 75%, 81%, 86%, 85%, 86%, 76%, 77%, 88%, 83%, 78%, 81%, 85%, 88%, and 79%, respectively.

[0047] Compound 1: 11H NMR (600 MHz, DMSO-D6) δ 9.36 (t, J = 5.6 Hz, 1H), 8.40 (s, 1H), 8.14 (s, 2H), 7.88 - 7.87 (d, J = 8.0 Hz, 1H), 7.53 - 7.51 (m, 1H), 7.44 (t, J = 7.7 Hz, 2H), 7.36 - 7.33 (m, 2H), 7.30 - 7.27 (m, 1H), 7.18 - 7.15 (m, 1H), 7.09 - 7.06 (m, 1H), 6.99 - 6.97 (m, 2H), 6.90 - 6.88 (dd, J = 8.0 Hz, J = 1.1 Hz, 1H), 4.53 - 4.52 (d, J = 5.6 Hz, 2H). 13 13C NMR (150 MHz, DMSO-D6) δ 161.81, 157.20, 153.85, 141.33, 139.67, 136.54, 130.37, 130.09, 130.08, 130.07, 130.07, 129.06, 128.71, 126.54, 124.09, 124.04, 123.73, 123.19, 120.76, 119.58, 119.15, 117.94, 117.93, 117.92, 48.70. ESI-MS m / z: 426.1273 [M+H] + .

[0048] Compound 2: 1 1H NMR (600 MHz, DMSO-D6) δ 13.20 (s, 1H), 9.45 (t, J = 5.6 Hz, 1H), 8.39 (s, 1H), 8.13 (s, 2H), 7.89 - 7.88 (d, J = 8.0 Hz, 1H), 7.52 - 7.51 (d, J = 7.3 Hz,​C NMR (150MHz, DMSO-D6) δ161.71, 156.88, 156.54, 141.73, 141.34, 139.66, 136.53, 130.39, 130.14, 130.13, 130.12, 130.11, 126.57, 123.97, 123.76, 123.60, 122.41, 120.77, 119.55, 118.80, 118.79, 118.78, 117.45, 117.05, 42.48. ESI-MS m / z: 426.1271[M+H] + .

[0049] Compound 3: 1 H NMR (600MHz, DMSO-D6) δ9.45 (t, J=5.5Hz, 1H), 8.41 (s, 1H), 8.14 (s, 2H), 7.89-7.88 (d, J=8.0Hz, 1H), 7.52-7.51 (d, J=7 .5Hz, 1H), 7.46-7.43 (m, 1H), 7.37-7.35 (m, 4H), 7.11 (t, J = 7.5Hz, 1H), 7.00-6.97 (m, 4H), 4.50-4.49 (d, J = 5.5Hz, 2H). 13 CNMR(150MHz,DMSO-D6)δ161.63,156.96,155.60,141.34,139.83,136.56,134.52,130.38,130.12,130.11,130.11 ,129.25,129.25,126.55,123.92,123.76,123.39,120.78,119.56,118.88,118.87,118.46,118.45,118.44,42.23. ESI-MS m / z: 426.1271[M+H] + .

[0050] Compound 4: 11H NMR (600 MHz, DMSO-D6) δ 9.22 (t, J = 5.9 Hz, 1H), 8.13 (s, 2H), 8.10 (d, J = 1.5 Hz, 1H), 8.05 (s, 1H), 7.95 - 7.94 (d, J = 8.5 Hz, 1H), 7.70 - 7.69 (dd, J = 8.5 Hz, J = 1.5 Hz, 1H), 7.40 - 7.38 (m, 1H), 7.36 - 7.33 (m, 2H), 7.27 - 7.24 (m, lH), 7.15 - 7.13 (m, 1H), 7.09 - 7.06 (m, 1H), 6.97 - 6.95 (m, 2H), 6.87 - 6.85 (m, 1H), 4.48 - 4.47 (d, J = 5.9 Hz, 2H). 13 13C NMR (150 MHz, DMSO-D6) δ 161.75, 157.14, 153.81, 140.29, 139.97, 137.84, 130.10, 130.10, 130.09, 130.07, 128.92, 128.65, 124.91, 124.42, 124.03, 123.21, 123.17, 120.97, 120.80, 119.05, 117.98, 117.97, 117.97, 117.96, 37.84. ESI-MS m / z: 426.1271 [M+H] + . [[ID=**6**]]

[0051] Compound 5: 1 1H NMR (600 MHz, DMSO-D6) δ 12.99 (s, 1H), 9.35 (t, J = 5.7 Hz, 1H), 8.26 - 8.06 (m, 4H), 7.98 - 7.97 (d, J = 8.5 Hz, 1H), 7.73 - 7.72 (dd, J = 8.5 Hz, J = 1.6 Hz, 1H), 7.38 - 7.34 (m, 3H), 7.13 - 7.10 (m, 2H), 7.01 - 6.99 (m, 3H), 6.88 - 6.87 (m, 1H), 4.48 - 4.47 (d, J = 5.7 Hz, 2H). 13C NMR (150MHz, DMSO-D6) δ161.66,156.83,156.60,141.75,140.27,139.94,137.84,130.14,130.13,130.13,130.09, 124.87,124.45,123.57,123.18,122.44,120.95,120.81,118.75,118.74,118.73,118.72,117.52,117.06,42.50. ESI-MS m / z: 426.1275[M+H] + .

[0052] Compound 6: 1 H NMR (600MHz, DMSO-D6) δ12.94(s,1H),9.31(t,J=5.7Hz,1H),8.23-8.06(m,4H),7.95-7.94(d,J=8.5Hz,1H ),7.70-7.69(m,1H),7.35-7.32(m,4H),7.09-7.07(m,1H),6.98-6.94(m,4H),4.45-4.44(d,J=5.7Hz,2H). 13 C NMR (150MHz, DMSO-D6) δ161.57,156.98,155.56,140.43,139.98,137.84,134.56,130.13,130.11,130.10,129.25, 129.24,124.83,124.43,123.38,123.18,120.96,120.80,118.88,118.87,118.86,118.43,118.43,118.43,42.24. ESI-MS m / z: 426.1269[M+H] + .

[0053] Compound 7: 11H NMR (600 MHz, DMSO-D6) δ 13.01 (s, 1H), 9.19 (t, J = 5.8 Hz, 1H), 8.36 - 8.20 (m, 2H), 8.07 (s, 2H), 7.89 - 7.88 (d, J = 8.2 Hz, 1H), 7.71 - 7.70 (dd, J = 8.2 Hz, J = 1.3 Hz, 1H), 7.42 - 7.36 (m, 3H), 7.30 - 7.27 (m, 1H), 7.18 - 7.09 (m, 2H), 6.99 - 6.98 (d, J = 8.2 Hz, 2H), 6.90 - 6.88 (d, J = 8.2 Hz, 1H), 4.51 - 4.50 (d, J = 5.8 Hz, 2H). 13 13C NMR (150 MHz, DMSO-D6) δ 161.80, 157. 16, 153.78, 141.28, 138.73, 137.27, 131.41, 130.12, 130.09, 130.08, 130.08, 128.93, 128.64, 125.55, 125.00, 124.03, 123.19, 122.99, 120.93, 119.06, 118.32, 117.95, 117.94, 117.94, 37.82. ESI-MS m / z: 426.1270 [M+H] + .

[0054] Compound 8: 1 1H NMR (600 MHz, DMSO-D6) δ 13.01 (s, 1H), 9.28 (t, J = 5.8 Hz, 1H), 8.31 (s, 1H), 8.26 (s, 1H), 8.06 - 8.04 (d, J = 8.4 Hz, 2H), 7.90 - 7.88 (d, J = 8.4 Hz, 1H), 7.71 - 7.69 (d, J = 8.4 Hz, 1H), 7.38 - 7.33 (m, 3H), 7.13 - 7.11 (m, 2H), 7.00 – 6.99 (d, J = 7.8 Hz, 3H), 6.88 - 6.87 (d, J = 7.8 Hz, 1H), 4.47 - 4.46 (d, J = 5.8 Hz, 2H). 13C NMR (150MHz, DMSO-D6) δ161.71,156.82,156.60,141.79,141.30,138.72,137.26,136.66,131.45,130.15,130.14, 130.09,126.09,125.59,124.97,123.57,123.02,122.45,120.93,118.73,118.73,118.33,117.54,117.05,42.49. ESI-MS m / z: 426.1273[M+H] + .

[0055] Compound 9: 1 H NMR (600MHz, DMSO-D6) δ13.01(s,1H),9.28(t,J=5.8Hz,1H),8.36-8.26(m,2H),8.08(s,2H),7.90-7.88(d,J=8.4Hz,1H),7 .71-7.69(dd,J=8.4Hz,J=1.3Hz,1H),7.38-7.35(m,4H),7.12-7.10(m,1H),7.00-6.97(m,4H),4.47-4.46(d,J=5.8Hz,2H). 13 C NMR (150MHz, DMSO-D6) δ161.62,156.99,155.54,141.29,138.88,137.28,134.60,131.44,130.12,130.11,130.10, 129.27,129.26,129.24,125.57,124.92,123.38,123.01,118.88,118.87,118.86,118.43,118.42,118.33,42.22. ESI-MS m / z: 426.1272[M+H] + .

[0056] Compound 10: 11H NMR (600 MHz, DMSO-D6) δ 9.25 (t, J = 5.6 Hz, 1H), 8.19 - 8.17 (m, 3H), 7.83 - 7.82 (d, J = 8.0 Hz, 1H), 7.68 - 7.67 (d, J = 7.5 Hz, 1H), 7.47 (t, J = 7.5 Hz, 1H), 7.44 - 7.42 (m, 1H), 7.39 - 7.35 (m, 2H), 7.30 - 7.27 (m, 1H), 7.18 - 7.15 (m, 1H), 7.11 - 7.09 (m, 1H), 7.00 - 6.98 (m, 2H), 6.90 - 6.88 (d, J = 8.0 Hz, 1H), 4.52 (d, J = 5.6 Hz, 2H). 13 13C NMR (150 MHz, DMSO-D6) δ 161.71, 157.17, 153.83, 140.14, 139.31, 137.51, 130.10, 130.09, 130.04, 130.02, 129.00, 128.68, 128.01, 125.72, 125.69, 124.33, 124.03, 123.33, 123.20, 119.44, 119.07, 118.00, 117.98, 117.93, 37.89. ESI-MS m / z: 426.1268 [M+H] + .

[0057] Compound 11: 1 1H NMR (600 MHz, DMSO-D6) δ 9.34 (t, J = 5.6 Hz, 1H), 8.19 - 8.16 (m, 3H), 7.84 - 7.82 (m, 1H), 7.68 - 7.67 (m, 1H), 7.47 (t, J = 7.5 Hz, 1H), 7.38 - 7.33 (m, 3H), 7.13 - 7.10 (m, 2H), 7.01 - 6.99 (m, 3H), 6.88 - 6.87 (m, 1H), 4.49 - 4.48 (d, J = 5.6 Hz, 2H). 13 13C NMR (150 MHz, DMSO-D6) δ 161.61, 156.82, 156.60, 141.70, 140.10, 139.28, 137.51, 13, 130.12, 130.08, 128.01, 125.69, 124.36, 123.57, 123.36, 122.46, 119.42, 118.74, 118.73, 118.73, 118.72, 117.54, 117.07, 42.51. ESI-MS m / z: 426.1271 [M+H] + .

[0058] Compound 12: 1 H NMR (600MHz, DMSO-D6) δ9.34(t,J=5.6Hz,1H),8.19(s,3H),7.84-7.82(d,J=8.0Hz,1H),7.68-7.67(d,J=7.6Hz, 1H),7.47(t,J=7.6Hz,1H),7.38-7.35(m,4H),7.12-7.10(m,1H),7.01-6.97(m,4H),4.49-4.48(d,J=5.6Hz,2H). 13 C NMR (150MHz, DMSO-D6) δ161.53,156.98,155.57,140.13,139.43,137.50,134.50,130.10,130.10,130.09,129.29, 129.28,128.02,125.69,125.65,124.33,123.37,123.34,119.42,118.86,118.85,118.42,118.42,118.41,42.26. ESI-MS m / z: 426.1272[M+H] + .

[0059] Compound 13: 1 H NMR(600MHz,DMSO-D6)δ9.29(t,J=5.8Hz,1H),8.90-8.89(d,J=6.2Hz,2H),8.20- 8.18(m,2H),7.94-7.93(d,J=6.2Hz,2H),7.63-7.57(m,2H),7.40-7.39(dd,J=7.6 Hz,J=1.3Hz,1H),7.34-7.31(m,2H),7.29-7.26(m,1H),7.16-7.13(m,1H),7.06(t ,J=7.6Hz,1H),6.95-6.94(m,2H),6.88-6.87(m,1H),4.50-4.49(d,J=5.8Hz,2H). 13CNMR(150MHz,DMSO-D6)δ161.46,157.18,153.86,151.02,146.77,141.49,136.76,134.61,130.05,130.04,130.03,130.02 ,129.93,129.17,128.77,126.51,125.94,125.29,124.39,124.04,123.15,122.94,119.18,117.91,117.90,117.89,37.97. ESI-MS m / z: 437.1319[M+H] + .

[0060] Compound 14: 1 H NMR (600MHz, DMSO-D6) δ9.39 (t, J=5.8Hz, 1H), 8.91-8.90 (d, J=6.0Hz, 2H), 8.23-8.19 (m, 2H), 7.97-7.96 (d, J=6.0Hz, 2H), 7.64-7.58 (m,2H),7.36-7.32(m,3H),7.11-7.08(m,2H),6.99-6.97(m,3H),6.87-6.85(dd,J=8.0Hz,J=2.1Hz,1H),4.47-4.46(d,J=5.8Hz,2H). 13 C NMR (150MHz, DMSO-D6) δ161.40,156.84,156.54,151.33,146.51,141.58,141.53,141.50,136.75,134.55,130.14,130.13, 130.12,130.11,126.55,126.01,125.41,124.47,123.59,122.90,122.49,118.76,118.75,118.74,117.56,117.09,42.51. ESI-MS m / z: 437.1321[M+H] + .

[0061] Compound 15: 11H NMR (600 MHz, DMSO-D6) δ 9.38 (t, J = 5.8 Hz, 1H), 8.89 - 8.88 (d, J = 6.0 Hz, 2H), 8.24 (s, 1H), 8.20 - 8.19 (d, J = 8.0 Hz, 1H), 7.93 - 7.92 (d, J = 6.0 Hz, 2H), 7.63 - 7.57 (m, 2H), 7.37 - 7.33 (m, 4H), 7.11 (t, J = 7.1 Hz, 1H), 6.98 - 6.95 (m, 4H), 4.47 - 4.46 (d, J = 5.8 Hz, 2H). 13 13C NMR (150 MHz, DMSO-D6) δ 161.31, 156.92, 155.64, 150.81, 146.97, 141.60, 141.50, 136.79, 134.69, 134.35, 130.11, 130.11, 130.10, 129.33, 129.31, 126.52, 125.93, 125.23, 124.34, 123.41, 122.86, 118.84, 118.83, 118.46, 118.46, 118.45, 42.24. ESI-MS m / z: 437.1319 [M+H] + .

[0062] Compound 16: 1 1H NMR (600 MHz, DMSO-D6) δ 9.37 (t, J = 5.8 Hz, 1H), 8.84 - 8.83 (d, J = 5.6 Hz, 2H), 8.52 (d, J = 1.7 Hz, 1H), 8.25 - 8.23 (m, 2H), 8.18 - 8.17 (m, 2H), 8.00 - 7.98 (dd, J = 8.5 Hz, J = 1.7 Hz, 1H), 7.44 - 7.42 (m, 1H), 7.39 - 7.36 (m, 2H), 7.31 - 7.28 (m, 1H), 7.19 - 7.16 (m, 1H), 7.11 - 7.09 (m, 1H), 7.00 - 6.98 (m, 2H), 6.90 - 6.89 (m, 1H), 4.53 - 4.52 (d, J = 5.8 Hz, 2H). 13C NMR (150MHz, DMSO-D6) δ161.46,157.14,153.84,146.48,142.15,141.50,140.06,132.75,130.11,130.09,130.08,130.08, 129.94,128.99,128.72,125.25,125.08,124.40,124.03,124.01,123.22,122.79,119.07,117.98,117.97,117.96,37.93. ESI-MSm / z: 437.1317[M+H] + .

[0063] Compound 17: 1 H NMR (600MHz, DMSO-D6) δ9.48 (t, J=5.7Hz, 1H), 8.87-8.86 (d, J=4.7Hz, 2H), 8.55 (s, 1H), 8.25-8.22 (m, 4H), 8.01-8. 00(m,1H),7.38-7.34(m,3H),7.13-7.10(m,2H),7.01-6.99(m,3H),6.89-6.87(m,1H),4.45-4.49(d,J=5.7Hz,2H). 13 C NMR (150MHz, DMSO-D6) δ161.37,156.84,156.59,151.95,145.73,142.36,141.63,141.55,140.05,132.49,130.13,130.13, 130.13,130.11,125.23,125.13,124.59,124.08,123.58,123.02,122.47,118.74,118.72,118.72,117.55,117.10,42.56. ESI-MS m / z: 437.1317[M+H] + .

[0064] Compound 18: 1 H NMR (600MHz, DMSO-D6) δ9.47(t,J=5.7Hz,1H),8.86-8.85(d,J=6.4Hz,2H),8.54(d,J=1.7Hz,1H),8.25-8.21(m,4H),8.0 1-7.99(dd,J=8.5Hz,J=1.7Hz,1H),7.38-7.35(m,4H),7.13-7.10(m,1H),7.01-6.96(m,4H),4.50-4.49(d,J=5.7Hz,2H).13 CNMR(150MHz,DMSO-D6)δ161.29,156.96,155.61,151.81,145.87,142.32,141.71,140.08,134.43,132.54,130.11,130.10 ,130.00,129.30,129.29,125.20,125.11,124.56,124.08,123.41,122.98,118.87,118.84,118.46,118.44,118.44,42.30. ESI-MS m / z: 437.1319[M+H] + .

[0065] Compound 19: 1 H NMR(600MHz,DMSO-D6)δ9.34(t,J=5.2Hz,1H),8.86-8.85(d,J=5.4Hz,2H),8.70(s,1H), 8.21-8.19(m,3H),8.13-8.11(m,1H),7.99-7.98(d,J=8.3Hz,1H),7.44-7.43(d,J=7.6Hz ,1H),7.39-7.36(m,2H),7.29(t,J=7.6Hz,1H),7.17(t,J=7.3Hz,1H),7.10(t,J=7.3Hz,1 H), 6.99-6.98 (d, J = 7.3Hz, 2H), 6.90-6.89 (d, J = 8.3Hz, 1H), 4.53-4.52 (d, J = 5.2Hz, 2H). 13 C NMR (150MHz, DMSO-D6) δ161.43,157.15,153.85,151.42,146.06,142.43,141.12,140.76,133.34,130.09,130.08,130.07, 129.91,129.07,128.74,126.17,124.70,124.15,124.03,123.20,122.91,122.41,119.09,117.97,117.96,117.95,37.97. ESI-MS m / z: 437.1317[M+H] + .

[0066] Compound 20: 11H NMR (600 MHz, DMSO-D6) δ 9.45 (t, J = 5.6 Hz, 1H), 8.86 - 8.85 (d, J = 6.0 Hz, 2H), 8.70 (s, 1H), 8.22 - 8.21 (d, J = 6.0 Hz, 2H), 8.19 (s, 1H), 8.14 - 8.12 (d, J = 8.3 Hz, 1H), 8.00 - 7.98 (dd, J = 8.3 Hz, J = 1.3 Hz, 1H), 7.39 - 7.34 (m, 3H), 7.13 - 7.11 (m, 2H), 7.01 - 6.99 (m, 3H), 6.89 - 6.87 (dd, J = 8.0 Hz, J = 2.2 Hz, 1H), 4.49 - 4.48 (d, J = 5.6 Hz, 2H). 13 13C NMR (150 MHz, DMSO-D) δ 161.37, 156.85, 156.59, 151.57, 145.94, 142.45, 141.59, 141.14, 140.78, 133.33, 130.16, 130.15, 130.15, 130.12, 126.22, 124.70, 124.20, 123.60, 122.97, 122.51, 122.47, 118.76, 118.75, 118.74, 117.58, 117.11, 42.58. ESI-MS m / z: 437.1316 [M+H] + .

[0067] Compound 21: 1 1H NMR (600 MHz, DMSO-D6) δ 9.45 (t, J = 5.4 Hz, 1H), 8.88 - 8.87 (d, J = 6.0 Hz, 2H), 8.71 (s, 1H), 8.25 - 8.20 (m, 3H), 8.14 - 8.11 (m, 1H), 8.00 - 7.98 (m, 1H), 7.38 - 7.34 (m, 4H), 7.11 (t, J = 7.6 Hz, 1H), 7.00 - 6.97 (m, 4H), 4.49 - 4.48 (d, J = 5.4 Hz, 2H). 13CNMR (150MHz, DMSO-D6) δ161.28, 156.96, 155.64, 152.06, 145.50, 142.74, 141.15, 140.92, 134.40, 133.15, 130.15, 130.13 , 130.11, 129.35, 129.34, 126.23, 124.67, 124.23, 123.42, 123.14, 122.57, 118.87, 118.86, 118.49, 118.47, 118.46, 42.35. ESI-MS m / z: 437.1319[M+H] + .

[0068] Compound 22: 1 H NMR (600MHz, DMSO-D6) δ9.28 (t, J=5.5Hz, 1H), 8.76-8.75 (d, J=5.4Hz, 2H), 8.22 (s, 1H) , 8.05-8.04 (dd, J=7.5Hz, J=1.3Hz, 1H), 7.78-7.77 (m, 2H), 7.65-7.60 (m, 2H), 7.42-7.4 0(dd, J=7.5Hz, J=1.3Hz, 1H), 7.38-7.35(m, 2H), 7.30-7.27(m, 1H), 7.18-7.15(m, 1H), 7 .09 (t, J=7.5Hz, 1H), 6.99-6.97 (m, 2H), 6.89-6.88 (m, 1H), 4.51-4.50 (d, J=5.5Hz, 2H). 13 C NMR (150MHz, DMSO-D6) δ161.43, 157.15, 153.83, 150.39, 150.38, 150.37, 146.90, 140.47, 139.98, 138.46, 133.29, 130.09, 130.08, 129.92, 129.01, 128.71, 126.43, 126.07, 125.63, 124.01, 123.19, 122.73, 119.07, 117.97, 117.96, 117.94, 37.91. ESI-MSm / z: 437.1321[M+H] + .

[0069] Compound 23: 11H NMR (600 MHz, DMSO-D6) δ 9.44 (t, J = 5.6 Hz, 1H), 8.91 - 8.90 (d, J = 5.7 Hz, 2H), 8.25 (s, 1H), 8.11 - 8.07 (m, 3H), 7.73 - 7.72 (d, J = 7.4 Hz, 1H), 7.64 (t, J = 7.4 Hz, 1H), 7.38 - 7.33 (m, 3H), 7.12 - 7.10 (m, 2H), 7.00 - 6.98 (m, 3H), 6.88 - 6.86 (m, 1H), 4.49 - 4.48 (d, J = 5.6 Hz, 2H). 13 13C NMR (150 MHz, DMSO-D6) δ 161.29, 156.83, 156.60, 150.43, 147.32, 141.57, 140.67, 140.17, 138.47, 132.25, 130.14, 130.13, 130.12, 130.09, 127.06, 126.96, 126.17, 125.55, 123.98, 123.57, 122.46, 118.72, 118.71, 118.70, 117.55, 117.11, 42.54. ESI-MS m / z: 437.1315 [M+H] + .

[0070] Compound 24: 1 1H NMR (600 MHz, DMSO-D6) δ 9.42 (t, J = 5.5 Hz, 1H), 8.89 - 8.88 (d, J = 5.8 Hz, 2H), 8.27 (s, 1H), 8.11 - 8.09 (d, J = 7.7 Hz, 1H), 8.04 - 8.03 (d, J = 6.0 Hz, 2H), 7.72 - 7.71 (d, J = 7.7 Hz, 1H), 7.64 (t, J = 7.7 Hz, 1H), 7.37 - 7.35 (m, 4H), 7.12 - 7.10 (m, 1H), 6.99 - 6.96 (m, 4H), 4.48 - 4.77 (d, J = 5.5 Hz, 2H). 13C NMR (150MHz, DMSO-D6) δ161.21, 156.96, 155.61, 150.02, 147.68, 140.68, 140.31, 138.47, 134.38, 132.38, 130.11, 130.11, 130.10, 129.29, 129.28, 126.97, 126.84, 126.16, 125.52, 123.83, 123.40, 118.86, 118.85, 118.44, 118.44, 118.42, 42.29. ESI-MSm / z: 437.1319[M+H] + .

[0071] Example 2

[0072] The synthetic route for benzothiophene derivatives in this embodiment is as follows: Figure 2 As shown, the specific preparation method is as follows:

[0073] Step 1, Synthesis of 6a-1:

[0074] Accurately weigh the halobenzothiophene-2-carboxylic acid (i.e. Figure 2 1b-c (1.00 g, 3.89 mmol), amines with the corresponding M5 group (i.e. Figure 2 5a-f (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 compound 6a-1 in yields of 89%, 85%, 85%, 86%, 82%, 81%, 86%, 78%, 84%, 86%, 82%, and 86%, respectively.

[0075] The second step is to synthesize 7a-l(25-36):

[0076] A mixture of the corresponding compound 6a-l (0.50 mmol), compound 1H-pyrazole-4-boronic acid (X1) (0.11 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 subjected to 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 25-36 with yields of 88%, 84%, 81%, 76%, 87%, 81%, 83%, 79%, 85%, 85%, 78%, and 83%, respectively.

[0077] Compound 25: 1 H NMR (600MHz, DMSO-D6) δ9.42 (t, J=6.0Hz, 1H), 8.16-8.15 (m, 3H), 8.09 (s, 1H), 7.99-7.98 (d, J=8.4Hz, 1H), 7.74-7.73 (d, J=8. 4Hz, 1H), 7.50-7.47 (m, 1H), 7.39-7.38 (d, J=7.8Hz, 1H), 7.32 (s, 1H), 7.26-7.25 (d, J=7.8Hz, 1H), 4.55-4.54 (d, J=6.0Hz, 2H). 13 C NMR (150MHz, DMSO-D6) δ161.80, 148.57, 142.35, 140.10, 139.95, 137.87, 130.48, 130.47, 130 .16, 126.47, 125.03, 124.51, 123.21, 121.03, 120.96, 120.86, 119.78, 119.51, 119.34, 42.25. ESI-MSm / z: 418.0832[M+H] + .

[0078] Compound 26: 1H NMR (600MHz, DMSO-D6) δ9.38 (t, J=5.7Hz, 1H), 8.17 (s, 2H), 8.14 (s, 1H), 8.10 (s, 1H), 7.99-7.98 (d, J=8.4Hz, 1H), 7.74- 7.72 (d, J=8.4Hz, 1H), 7.41-7.34 (m, 1H), 7.23-7.22 (m, 2H), 7.15 (s, 1H), 7.10-7.07 (m, 1H), 4.51-4.50 (d, J=5.9Hz, 2H). 13 CNMR (150MHz, DMSO-D6) δ161.72, 151.10, 141.87, 140.21, 139.96, 137.87, 130.15, 130.12, 124 .96, 124.48, 124.19, 123.19, 121.05, 120.84, 118.14, 117.82, 117.22, 116.43, 114.72, 42.40. ESI-MSm / z: 400.0926[M+H] + .

[0079] Compound 27: 1 H NMR (600MHz, DMSO-D6) δ13.00(s,1H),9.32(t,J=5.8Hz,1H),8.14(s,2H),8.09(s,2H),7.99-7.97(d,J=8.3Hz,1H),7.73-7.72(d,J=8.3Hz,1 H),7.43(s,1H),7.42(s,1H),7.36-7.34(m,2H),7.30-7.24(m,2H),6. 99(s,1H),6.93-6.89(m,2H),5.08(s,2H),4.47-4.46(d,J=5.8Hz,2H). 13 C NMR (150MHz, DMSO-D6) δ161.60,158.52,141.02,140.41,139.98,137.85,137.14,130.11,129.56,128.49,128.48,128 .46,127.88,127.78,127.78,127.77,124.83,124.42,123.17,120.96,120.81,119.78,114.04,113.10,69.20,42.66. ESI-MS m / z: 440.1428[M+H] + .

[0080] Compound 28: 11H NMR (600 MHz, DMSO-D6) δ 12.98 (s, 1H), 9.32 (t, J = 5.9 Hz, 1H), 8.12 - 8.10 (m, 3H), 8.03 (s, 1H), 7.95 - 7.94 (d, J = 8.3 Hz, 1H), 7.70 - 7.69 (m, 1H), 7.39 - 7.37 (m, 2H), 7.34 (t, J = 7.9 Hz, 1H), 7.12 - 7.11 (d, J = 7.9 Hz, 1H), 7.00 - 6.98 (m, 3H), 6.89 - 6.87 (dd, J = 8.0 Hz, J = 2.3 Hz, 1H), 4.46 - 4.45 (d, J = 5.9 Hz, 2H). 13 13C NMR (150 MHz, DMSO-D6) δ 161.67, 156.43, 155.63, 141.91, 140.24, 139.93, 137.84, 130.22, 130.12, 129.99, 129.98, 129.96, 127.30, 124.89, 124.46, 123.18, 122.85, 120.96, 120.82, 120.36, 120.35, 120. 34, 117.67, 117.29, 42.45. ESI-MS m / z: 460.0882 [M + H] + .

[0081] Compound 29: 1 1H NMR (600 MHz, DMSO-D6) δ 12.96 (s, 1H), 9.34 (t, J = 5.8 Hz, 1H), 8.16 - 8.13 (s, 3H), 8.06 (s, 1H), 7.98 - 7.97 (d, J = 8.5 Hz, 1H), 7.73 - 7.72 (d, J = 8.5 Hz, 1H), 7.3 4 (t, J = 7.7 Hz, 1H), 7.23 - 7.19 (m, 2H), 7.11 - 7.10 (d, J = 7.7 Hz, 1H), 7.07 - 7.04 (m, 2H), 6.97 (s, 1H), 6.86 - 6.84 (m, 1H), 4.47 - 4.46 (d, J = 5.8 Hz, 2H). 13C NMR (150MHz, DMSO-D6) δ161.66,159.13,157.54,157.32,152.50,141.78,140.26,139.93,137.83,130.12,130.10, 124.87,124.45,123.17,122.30,120.96,120.87,120.82,120.81,120.80,116.98,116.75,116.60,116.50,42.48. ESI-MS m / z: 444.1177[M+H] + .

[0082] Compound 30: 1 H NMR (600MHz, DMSO-D6) δ9.37(t,J=5.9Hz,1H),8.15(s,2H),8.13(s,1H),8.07(s,1H),7.98-7.97(d,J=8.5Hz,1H),7.73-7.71 (m,3H),7.43(t,J=7.8Hz,1H),7.23-7.22(d,J=7.8Hz,1H),7.14-7.11(m,3H),7.03-7.01(m,1H),4.52-4.51(d,J=5.9Hz,2H). 13 C NMR (150MHz, DMSO-D6) δ161.70,160.36,155.18,142.17,140.22,139.93,137.86,130.44,130.10,127.57,127.55,125 .26,124.92,124.47,123.74,123.53,123.46,123.32,123.17,120.97,120.82,118.74,118.44,118.07,118.05,42.42. ESI-MS m / z: 494.1145[M+H] + .

[0083] Compound 31: 1H NMR (600MHz, DMSO-D6) δ13.01(s,1H),9.35(t,J=6.0Hz,1H),8.27(s,2H),8.09(s,2H),7.91-7.90(d,J=8.4Hz,1H),7.72-7.70(d,J= 8.4Hz, 1H), 7.48 (t, J = 7.7Hz, 1H), 7.38-7.37 (d, J = 7.7Hz, 1H), 7.32 (s, 1H), 7.26-7.25 (d, J = 7.7Hz, 1H), 4.54-4.53 (d, J = 6.0Hz, 2H). 13 C NMR (150MHz, DMSO-D6) δ161.84,148.55,142.38,141.33,138.53,137.25,131.51,130.46,130 .45,126.48,125.63,125.13,123.05,121.03,120.92,119.79,119.48,119.33,118.34,42.25. ESI-MS m / z: 418.0832[M+H] + .

[0084] Compound 32: 1 H NMR (600MHz, DMSO-D6) δ9.32(t,J=5.9Hz,1H),8.27(s,1H),8.18(s,2H),8.09(s,1H),7.91-7.89(d,J=8.4Hz,1H),7.72-7.70(d,J =8.4Hz,1H),7.63-7.52(m,1H),7.41-7.34(m,1H),7.23-7.21(m,1H),7.14(s,1H),7.10-7.06(m,1H),4.50-4.49(d,J=5.9Hz,2H). 13 C NMR (150MHz, DMSO-D6) δ161.76,151.11,141.90,141.32,138.65,137.26,131.47,130.11,128 .81,125.59,125.05,124.20,123.03,120.92,118.33,118.14,117.20,116.43,114.72,42.39. ESI-MS m / z: 400.0926[M+H] + .

[0085] Compound 33: 11H NMR (600 MHz, DMSO-D6) δ 13.01 (s, 1H), 9.26 (t, J = 6.0 Hz, 1H), 8.31 (s, 1H), 8.26 (s, 1H), 8.08 (s, 1H), 8.05 (s, 1H), 7.90 - 7.89 (d, J = 8.3 Hz, 1H), 7.71 - 7.70 (d, J = 8.3 Hz, 1H), 7.43 (s, 1H), 7.42 (s, 1H), 7.35 (t, J = 7.6 Hz, 2H), 7.30 - 7.24 (m, 2H), 6.99 (s, 1H), 6.92 - 6.89 (m, 2H), 5.08 (s, 2H), 4.46 - 4.45 (d, J = 6.0 Hz, 2H). 13 13C NMR (150 MHz, DMSO-D6) δ 161.65, 158.52, 141.29, 141.05, 138.85, 137.28, 137.14, 131.41, 129.54, 128.50, 128.48, 128.48, 127.88, 127.78, 127.77, 127.77, 125.56, 124.92, 123.00, 120.93, 119.79, 118.32, 114.04, 113.08, 69.20, 42.65. ESI-MS m / z: 440.1428 [M+H] + .

[0086] Compound 34:<00001​​C NMR (150MHz, DMSO-D6) δ161.72,156.41,155.64,141.94,141.29,138.68,137.24,131.45,130.20,129.99,129.97, 129.97,127.29,125.57,124.98,123.01,122.87,120.92,120.34,120.34,120.33,118.32,117.70,117.28,42.44. ESI-MS m / z: 460.0882[M+H] + .

[0087] Compound 35: 1 H NMR (600MHz, DMSO-D6) δ13.02 (s, 1H), 9.28 (t, J = 5.8Hz, 1H), 8.26-8.12 (s, 3H),8.05(s,1H),7.90-7.88(d,J=8.5Hz,1H),7.71-7.70(d,J=8.5Hz,1H), 7.34(t,J=7.7Hz,1H),7.23-7.19(m,2H),7.11-7.09(d,J=7.8Hz,1H),7.07 -7.04(m,2H),6.97(s,1H),6.85-6.84(m,1H),4.47-4.46(d,J=5.8Hz,2H). 13 C NMR (150MHz, DMSO-D6) δ161.70,159.12,157.53,157.30,152.51,141.81,141.29,138.70,137.24,131.44,130.10, 130.09,125.57,124.96,123.01,122.31,120.92,120.86,120.80,118.32,117.00,116.75,116.59,116.48,42.47. ESI-MS m / z: 444.1177[M+H] + .

[0088] Compound 36: 1H NMR (600MHz, DMSO-D6) δ9.30 (t, J = 5.9Hz, 1H), 8.26 (s, 1H), 8.18 (s, 2H), 8.06 (s, 1H), 7.89-7.88 (d, J = 8.2Hz, 1H), 7.72-7.69 (m, 3H), 7.42 (t, J=7.6Hz, 1H), 7.23-7.21 (d, J=7.6Hz, 1H), 7.14-7.11 (m, 3H), 7.02-7.01 (m, 1H), 4.51-4.50 (d, J=5.9Hz, 2H). 13 C NMR (150MHz, DMSO-D6) δ161.75, 160.37, 155.16, 142.21, 141.30, 138.66, 137.24, 131.45, 130.42, 127.57, 127.55, 125 .57, 125.26, 125.01, 123.75, 123.52, 123.46, 123.31, 123.02, 120.92, 118.77, 118.43, 118.32, 118.06, 118.05, 42.41. ESI-MSm / z: 494.1145[M+H] + .

[0089] Example 3

[0090] The synthetic route for benzothiophene derivatives in this embodiment is as follows: Figure 3 As shown, the specific preparation method is as follows:

[0091] Step 1, synthesize 8a-b:

[0092] Accurately weigh the halobenzothiophene-2-carboxylic acid (i.e. Figure 3 Compounds 1b-c (1.00 g, 3.89 mmol), 3-(aminomethyl)phenol (0.72 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, followed by stirring at room temperature for 24 hours. 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 purified by silica gel column chromatography with petroleum ether / ethyl acetate (1:1, v / v, 500 mL) to give compounds 8a-b in yields of 90% and 88%, respectively.

[0093] The second step is to synthesize 9a-d:

[0094] A mixture of the corresponding compounds 8a-b (1 mmol), 2-bromo-N,N-dimethylethylamine (0.31 g, 2 mmol) or 3-bromo-N,N-dimethylpropylamine (0.33 g, 2 mmol) and anhydrous cesium carbonate (1.42 g, 4 mmol) was stirred at 90 °C for 20 hours under a nitrogen atmosphere with anhydrous dimethylformamide as the solvent. After cooling to room temperature, the solvent was removed by concentration under reduced pressure. The residue was extracted with dichloromethane, and the extraction 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 with petroleum ether / ethyl acetate (1:2, v / v, 500 mL) to give compounds 9a-d in yields of 65%, 68%, 65%, and 71%, respectively.

[0095] The third step is to synthesize 10a-d(37-40):

[0096] A mixture of the corresponding compounds 9a-d (0.50 mmol), compound 1H-pyrazole-4-boronic acid (X1) (0.11 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 subjected to high-performance liquid chromatography (HPLC) (separation conditions: preparative column, Welch). XB-C18, 21.2×250mm, 5μm; elution system: methanol / water (volume ratio) = 10% v 100%, containing 0.5‰ trifluoroacetic acid (volume percentage), flow rate: 10ml / min, elution time: 40min) purification, yielding the corresponding compounds 37-40, with yields of 78%, 81%, 88%, and 79%, respectively.

[0097] Compound 37: 11H NMR (600 MHz, DMSO-D6) δ 9.69 (s, 1H), 9.38 (t, J = 6.0 Hz, 1H), 8.16 (s, 2H), 8.13 (d, J = 1.6 Hz, 1H), 8.10 (s, 1H), 7.99 - 7.97 (d, J = 8.5 Hz, 1H), 7.74 - 7.72 (dd, J = 8.5 Hz, J = 1.6 Hz, 1H), 7.30 (t, J = 7.8 Hz, 1H), 6.99 - 6.96 (m, 2H), 6.91 - 6.89 (dd, J = 7.8 Hz, J = 2.8 Hz, 1H), 4.48 - 4.47 (d, J = 6.0 Hz, 2H), 4.28 (t, J = 5.0 Hz, 2H), 3.51 - 3.48 (m, 2H), 2.84 (d, J = 3.5 Hz, 6H). 13 13C NMR (150 MHz, DMSO-D6) δ 161.64, 158.34, 158.12, 157.75, 141.23, 140.37, 139.97, 137.83, 130.16, 129.67, 124.86, 124.48, 123.19, 120.93, 120.78, 120.44, 113.96, 112.81, 62.09, 55.60, 42.88, 42.86, 42.62. ESI-MS m / z: 421.1693 [M + H] + .

[0098] Compound 38: 1 1H NMR (600 MHz, DMSO-D6) δ 9.72 (s, ¹H), 9.37 (t, J = 6.0 Hz, 1H), 8.16 (s, 2H), 8.13 (s, 1H), 8.10 (s, 1H), 7.99 - 7.97 (d, J = 8.5 Hz, 1H), 7.74 - 7.72 (d, J = 8.5 Hz, 1H), 7.26 (t, J = 7.9 Hz, 1H), 6.94 - 6.93 (d, J = 7.9 Hz, 1H), 6.90 (s, 1H), 6.84 - 6.82 (dd, J = 7.9 Hz, J = 2.8 Hz, 1H), 4.47 - 4.46 (d, J = 6.0 Hz, 2H), 4.01 (t, J = 5.9 Hz, 2H), 3.32 - 3.19 (m, 2H), 2.80 - 2.79 (d, J = 3.9 Hz, 6H), 2.10 - 2.06 (m, 2H). 13C NMR (150MHz, DMSO-D6) δ161.64, 158.55, 158.43, 158.32, 141.10, 140.41, 140.00, 137.85, 130.15, 129.59, 1 24.86, 124.47, 123.19, 120.97, 120.81, 119.92, 113.75, 112.68, 64.70, 54.36, 42.67, 42.37, 42.37, 24.07. ESI-MS m / z: 435.1850[M+H] + .

[0099] Compound 39: 1 H NMR (600MHz, DMSO-D6) δ9.74 (s, 1H), 9.32 (t, J=5.8Hz, 1H), 8.26 (s, 1H), 8.18 (s, 2H) , 8.09 (s, 1H), 7.90-7.89 (d, J=8.4Hz, 1H), 7.72-7.70 (dd, J=8.4Hz, J=1.8Hz, 1H), 7.3 0 (t, J=7.8Hz, 1H), 6.99-6.96 (m, 2H), 6.90-6.89 (dd, J=7.8Hz, J=2.3Hz, 1H), 4.47-4 .46 (d, J=5.8Hz, 2H), 4.28 (t, J=5.0Hz, 2H), 3.51-3.48 (m, 2H), 2.84 (d, J=3.5Hz, 6H). 13 C NMR (150MHz, DMSO-D6) δ161.69, 158.34, 158.12, 157.75, 141.29, 141.26, 138.82, 137.27, 131.47, 129.6 6, 125.58, 124.96, 123.05, 120.91, 120.45, 118.32, 113.97, 112.80, 62.10, 55.59, 42.88, 42.87, 42.61. ESI-MS m / z: 421.1693[M+H] + .

[0100] Compound 40: 1H NMR (600MHz, DMSO-D6) δ9.59 (s, 1H), 9.30 (t, J=5.8Hz, 1H), 8.26 (s, 1H), 8.18 (s, 2H), 8.09 (s, 1H) , 7.90-7.89 (d, J=8.4Hz, 1H), 7.71-7.70 (dd, J=8.4Hz, J=1.6Hz, 1H), 7.26 (t, J=8.0Hz, 1H), 6.94-6 .93 (d, J=7.7Hz, 1H), 6.90-6.89 (m, 1H), 6.84-6.82 (dd, J=8.0Hz, J=2.3Hz, 1H), 4.46-4.44 (d, J=5. 8Hz, 2H), 4.01 (t, J=6.0Hz, 2H), 3.22-3.19 (m, 2H), 2.80-2.79 (d, J=4.6Hz, 6H), 2.10-2.05 (m, 2H). 13 C NMR (150MHz, DMSO-D6) δ162.12, 158.86, 158.74, 158.52, 141.74, 141.57, 139.28, 137.72, 131.91, 130.03, 1 26.02, 125.39, 123.48, 121.37, 120.37, 118.77, 114.19, 113.10, 65.13, 54.82, 43.10, 42.94, 42.83, 24.52. ESI-MS m / z: 435.1850[M+H] + .

[0101] Example 1: ROCK Activity Test

[0102] 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. KD025 was used as a positive control, 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 × 10⁴. The 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 1.

[0103] Table 1

[0104]

[0105]

[0106] Note: NT indicates untested.

[0107] Conclusion: Compounds 5, 8, 37, 38, 39, and 40 in this disclosure exhibit excellent inhibitory activity against ROCK2 kinase, and their ROCK2 / ROCK1 selectivity is stronger than that of the positive control drug KD025.

[0108] Example 2: Efficacy test of compound 40 in treating atopic dermatitis in female Balb / c mice.

[0109] Of the compounds 5, 8, 37, 38, 39, and 40 disclosed in this invention, compound 40 was investigated for its therapeutic effect in a calcipotriol (MC903)-induced mouse atopic dermatitis (AD) model, given its good activity and optimal selectivity. Specifically, MC903 dissolved in 95% ethanol (1 nmol, 20 μL) was topically applied to the inner side of the right ear of mice, while a control group received 20 μL of 95% ethanol in the same area. The selective ROCK2 inhibitor KD025 and the oral small molecule drug abrocitinib were used as positive controls. Abrocitinib is a JAK1 inhibitor that was approved globally in 2021 and in China in 2022 for the treatment of AD. Mice were divided into six groups: (1) Control group: mice received an equal volume of 95% ethanol; (2) Model group: mice were induced with atopic dermatitis (AD) using MC903; (3) Low-dose group: mice were induced with AD using MC903 and orally administered compound 40 (dose 25 mg / kg); (4) High-dose group: mice were induced with AD using MC903 and orally administered compound 40 (dose 50 mg / kg); (5) KD025 positive control group: mice were induced with AD using MC903 and orally administered KD025 (dose 50 mg / kg); (6) Abrocitinib positive control group: mice were induced with AD using MC903 and orally administered Abrocitinib (dose 50 mg / kg). After successfully establishing the MC903-induced AD model on day 7, the mice were administered the drug for seven consecutive days.

[0110] Conclusion: Figure 4 As shown, compound 40 in the disclosed compounds of this invention not only exhibits good activity and selectivity, but also demonstrates better efficacy in treating atopic dermatitis than the positive control drug KD025 and abuxitinib at the same dosage.

[0111] 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.

[0112] 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.

[0113] 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-based ROCK2 inhibitor, characterized in that, Compounds having the chemical structure shown in Formula I and / or their pharmaceutically acceptable salts: In Formula I, R1 is at least one of hydrogen atom, halogen group, pyrazolyl group, and pyridinyl group, and R2 is at least one of methoxyphenyl, trifluoromethoxy, difluoromethoxy, phenethoxy, 4-chloro-1-methoxyphenyl, 4-fluoro-1-methoxyphenyl, and 4-(trifluoromethyl)-1-methoxyphenyl. And / or, compounds having the chemical structure shown in Formula II and / or their pharmaceutically acceptable salts: In Formula II, R3 is at least one of a hydrogen atom, a halogen group, or a pyrazol group, and n is 2 or 3.

2. The benzothiophene-based ROCK2 inhibitor according to claim 1, characterized in that, Selected from at least one of the following compounds or their pharmaceutically acceptable salts:

3. The use of the benzothiophene ROCK2 inhibitor according to claim 1 or 2 in the preparation of a drug for inhibiting ROCK2.

4. The use of the benzothiophene ROCK2 inhibitor according to claim 1 or 2 in the preparation of a medicament for the treatment and / or prevention of diseases associated with abnormal ROCK2 activity.

5. The application according to claim 4, characterized in that, The disease described is inflammation and / or cancer.

6. The application according to claim 5, characterized in that, The cancers mentioned are at least one of the following: breast cancer, endometrial cancer, ovarian cancer, vaginal cancer, fallopian tube cancer, cervical cancer, kidney cancer, bladder cancer, urothelial carcinoma, urethral cancer, prostate cancer, testicular cancer, colorectal cancer, sarcoma, bone cancer, multiple myeloma, leukemia, myxoma, rhabdomyosarcoma, leiomyosarcoma, fibroma, lipoma, teratoma, pharyngeal cancer, nasopharyngeal cancer, oral cancer, lung cancer, alveolar carcinoma, lymphoma, mesothelioma, small intestine cancer, gastric cancer, esophageal cancer, pancreatic cancer, liver cancer, bile duct cancer, neurofibroma, glioma, neuroblastoma, neuroblastoma, melanoma, skin cancer, basal cell carcinoma, squamous cell carcinoma, thyroid cancer, head and neck cancer, salivary gland cancer, and gastrointestinal stromal tumor.

7. The application according to claim 5, characterized in that, The inflammation is at least one of the following: atopic dermatitis, enteritis, psoriasis, psoriatic arthritis, encephalitis, keratitis, conjunctivitis, rhinitis, otitis media, gingivitis, pharyngitis, tonsillitis, pneumonia, hepatitis, dysentery, prostatitis, endometritis, cervicitis, pelvic inflammatory disease, paronychia, myocarditis, rheumatoid arthritis, osteoarthritis, juvenile idiopathic arthritis, ankylosing spondylitis, allergic airway disease, chronic obstructive pulmonary disease, asthma, bronchitis, systemic lupus erythematosus, cutaneous lupus erythematosus, lupus nephritis, dermatomyositis, autoimmune liver disease, Sjögren's syndrome, multiple sclerosis, dry eye disease, diabetes and related complications, thyroiditis, contact dermatitis, and amyotrophic lateral sclerosis.

8. A pharmaceutical composition, characterized in that, Includes one or more of pharmaceutically acceptable carriers, diluents, and excipients, as well as the benzothiophene ROCK2 inhibitors as described in claim 1 or 2.

9. The method for preparing benzothiophene-based ROCK2 inhibitors according to claim 1 or 2, characterized in that, The benzothiophene-based ROCK2 inhibitor is a compound having the chemical structure shown in Formula I, and its preparation method includes the following steps: Synthetic route A: The product 3a-1 is prepared by amidation of halobenzothiophene-2-carboxylic acid and amine with the corresponding M2 group in the presence of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate; the product 3a-1 is then reacted with boric acid with the corresponding M3 group by a Suzuki reaction to obtain a compound with the chemical structure shown in Formula I. The M2 group is methoxyphenyl, and the M3 group is pyrazolyl or pyridinyl. or, Synthetic route B: The product 6a-1 is prepared by amidation of halobenzothiophene-2-carboxylic acid and amine with the corresponding M5 group in the presence of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate; the product 6a-1 is reacted with pyrazoleboronic acid by Suzuki reaction to obtain a compound with the chemical structure shown in Formula I. The M5 group is at least one of trifluoromethoxy, difluoromethoxy, phenethoxy, 4-chloro-1-methoxyphenyl, 4-fluoro-1-methoxyphenyl, and 4-(trifluoromethyl)-1-methoxyphenyl. Alternatively, the benzothiophene ROCK2 inhibitor is a compound having the chemical structure shown in Formula II, and its preparation method includes the following steps: Synthetic route C: Halogenated benzothiophene-2-carboxylic acid and 3-(aminomethyl)phenol undergo amidation in the presence of N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate to yield products 8a-b; product 8a-b undergoes nucleophilic substitution with 2-bromo-N,N-dimethylethylamine or 3-bromo-N,N-dimethylpropylamine in the presence of Cs2CO3 to generate products 9a-d; product 9a-d undergoes a Suzuki reaction with pyrazolboric acid to give a compound having the chemical structure shown in Formula II.

10. The method for preparing the benzothiophene-based ROCK2 inhibitor according to claim 9, characterized in that, In synthetic route A, the molar ratio of halobenzothiophene-2-carboxylic acid and amine having the corresponding M2 group is 1:1.5; a first catalyst is also added during the reaction; product 3a-1 and boric acid having the corresponding M3 group are dissolved in a solvent, and then a second catalyst and a base are added to carry out the reaction, wherein the molar ratio of product 3a-1 and boric acid having the corresponding M3 group is 1:2; In synthetic route B, the molar ratio of halobenzothiophene-2-carboxylic acid and amine having the corresponding M5 group is 1:1.5; a first catalyst is also added during the reaction; product 6a-1 and pyrazoleboronic acid are dissolved in a solvent, and then a second catalyst and a base are added to react, wherein the molar ratio of product 6a-1 and pyrazoleboronic acid is 1:

2. In synthetic route C, the molar ratio of halobenzothiophene-2-carboxylic acid and 3-(aminomethyl)phenol is 1:1.5; a first catalyst is also added during the reaction; the molar ratio of product 8a-b to 2-bromo-N,N-dimethylethylamine or 3-bromo-N,N-dimethylpropylamine is 1:2; product 9a-d and pyrazolonic acid are dissolved in a solvent, and then a second catalyst and a base are added to react, wherein the molar ratio of product 9a-d to pyrazolonic acid is 1:

2. The first catalyst is N-methylimidazole, the second catalyst is tetra(triphenylphosphine)palladium, the solvent is a mixed solution of dioxane and water, and the base is potassium carbonate.

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