4,5,6,7-Tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide compounds, their preparation methods and applications

Abstract

This invention discloses 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compounds, their preparation methods, and applications, belonging to the field of pharmaceutical technology. Specifically, the general structural formula of the 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compounds or their pharmaceutically acceptable salts is shown in Formula I, where R1 is defined in the claims and specification. The 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compounds or their pharmaceutically acceptable salts provided by this invention, as inhibitors of vascular endothelial growth factor receptor 2 (VEGFR-2) kinase and epidermal growth factor receptor (EGFR) kinase, can be used to prepare drugs for the treatment or prevention of tumors related to VEGFR-2 kinase and EGFR kinase, demonstrating good application value and promising development prospects.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compounds and their preparation methods, as well as their application as inhibitors of vascular endothelial growth factor receptor kinase and epidermal growth factor receptor kinase in the preparation of antitumor drugs. Background Technology

[0002] In today's world, malignant tumors remain the leading cause of death. With the development of molecular biology techniques, the pathogenesis of tumors is gradually being elucidated, and a series of different therapeutic targets are being discovered. Through rational drug design, small molecules that can target and bind to protein-protein interaction sites can promote or block the biological functions of related proteins, thereby achieving the goal of disease treatment. This is currently a hot topic in the field of tumor treatment.

[0003] Imatinib is the first targeted anti-tumor drug developed and manufactured by Novartis in Switzerland. It inhibits cancer cell growth by selectively targeting specific molecules involved in cancer cell growth (Druker, BJ; Talpaz, M.; Resta, DJ; Peng, B.; Buchdunger, E.; Ford, JM; Lydon, NB; Kantarjian, H.; Capdeville, R.; Ohno-Jones, S.; Sawyers, CLEfficacy and safety of a specific inhibitor of the BCR-ABL tyrosine kinase in chronic myeloid leukemia. New Engl J Med, 2001, 344, 1031-1037). In 2001, the U.S. Food and Drug Administration (FDA) approved imatinib for the treatment of chronic myeloid leukemia and gastrointestinal stromal tumors. Imatinib has been synthesized as a potential inhibitor of platelet-derived growth factor receptor (PDGFR) and c-kit receptor, and also has good activity against BCR-ABL protein.

[0004] Axitinib, developed by Pfizer and marketed under the brand name Inlyta, was approved in 2012 for the treatment of advanced renal cell carcinoma following failure of prior systemic therapy. Pre-approval clinical trials had been reviewed elsewhere. The development of resistance to VEGFR inhibitors in patients with metastatic advanced renal cell carcinoma led to its combination with PD-1 inhibitors such as avelumumab or pemetremumab. In 2019, it received approval (Grünwald V, Voss MH, Rini BI, et al. Axitinib plus immune checkpoint inhibitor: evidence-and expert-based consensus recommendation for treatment optimization and management of related adverse events. Br J Cancer, 2020, 123, 898-904). Axitinib primarily targets VEGFR-1, VEGFR-2, VEGFR-3, and PDGFRβ, and is a type IIA selective inhibitor of VEGFR with sub-millimolar potency.

[0005] Cabozantinib, developed by Exelixis and licensed to Bristol-Myers Squibb, is marketed as Cometriq (capsules) and Cabometyx (tablets). Cometriq was approved in 2012 for the treatment of patients with advanced metastatic medullary thyroid carcinoma. In 2016, Cabometyx was approved for the treatment of patients with advanced renal cell carcinoma who had previously received anti-angiogenic therapy. A year later, it was officially approved as a first-line treatment for patients with advanced renal cell carcinoma. Cabozantinib was approved in 2019 for the treatment of liver cancer patients who had previously received sorafenib. Its primary targets are MET, VEGFR-2, and RET. In biochemical assays, cabozantinib showed effective inhibition of VEGFR-2 and MET kinases, with an IC50 concentration of [missing information]. 50 In the picomol and nanomol ranges, respectively (Yakes FM, Chen J, Tan J, et al. Cabozantinib (XL184), a novel MET and VEGFR2 inhibitor, simultaneously suppresses metastasis, angiogenesis, and tumor growth. Mol Cancer Ther, 2011, 10, 2298-2308). Summary of the Invention

[0006] The object of this invention is to provide a compound as shown in Formula I, its prodrug and pharmaceutically active metabolite, and its pharmaceutically acceptable salt, and to provide a method for its preparation and its use in the preparation of treatments and / or preventions of tumor diseases associated with vascular endothelial growth factor receptor kinase and epidermal growth factor receptor kinase.

[0007]

[0008] R1 is substituted by 1-3 substituents selected from hydrogen, methyl, methoxy, ethyl, halogen, cyano, isobutyl, n-pentyl, and dimethylamino.

[0009] Furthermore, the 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compounds or pharmaceutically acceptable salts thereof described in this invention are selected from:

[0010] 6-(4-methoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide;

[0011] 6-(4-ethylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide;

[0012] 6-(3-methylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide;

[0013] 6-(4-chlorobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide;

[0014] 6-Benzyl-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide;

[0015] 6-(2,3-dimethoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide;

[0016] 6-(3,4,5-trimethoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide;

[0017] 6-(4-methylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureo}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3-cyanobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureo}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(4-isobutylbenzyl)-2-{3- [4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(4-pentylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3-ethylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl} -4,5,6,7-Tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(2-methoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(2-chlorobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3,5-difluorobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3,4-dimethoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide;

[0018] 6-(4-Dimethylaminobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureo}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3,5-dimethylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureo}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide Amine; 6-(2-methylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3,6-dichlorobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide.

[0019] The present invention also provides a pharmaceutical composition comprising, as an active ingredient, any one of the following compounds: a 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compound of the present invention, a prodrug and a pharmaceutically active metabolite thereof, and a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

[0020] "Pharmaceutically acceptable salt" refers to a conventional acid addition salt or base addition salt that retains the biological potency and properties of a compound of formula I and is formed with a suitable nontoxic organic or inorganic acid or organic or inorganic base. Acid addition salts include hydrochlorides, hydrobroms, hydroiodates, nitrates, phosphates, sulfates, perchlorates, thiocyanates, hydrogen sulfates, persulfates, borates, formates, acetates, propionates, valerates, neovalerates, hexanoates, heptanoates, octanoates, isooctanoates, undecanoates, laurates, palmitates, stearates, oleates, cyclopropionates, oxalates, malonates, succinates, maleates, fumarates, adipates, azelaates, acrylates, strawberry salts, crotonates, tigrinates, itacrates, sorbates, cinnamates, glycolates, lactates, malates, tartrates, citrates, tartrites, mandelates, diphenylglycolates, tropine, ascorbate, gluconate, glucono-p-ethyl, gluconate, mannitol, lactobionate, benzoates, phthalates, paraphthalates, furoate, nicotinic acid, and isonicotinic acid. Salicylate, acetylsalicylate, butyrate, gallate, caffeate, ferulic acid, picrate, camphorate, camphor sulfonate, methanesulfonate, ethanesulfonate, propanesulfonate, benzenesulfonate, p-toluenesulfonate, p-aminobenzenesulfonate, aminosulfonate, taurine, 2-hydroxyethanesulfonate, glycine, alanine, valine, leucine, isoleucine, phenylalanine, tryptophan, tyrosine, aspartate, asparagine, glutamate, lysine, glutamine, methionine, serine, threonine, cysteine, proline, histidine, arginine, edetate, pyruvate, α-ketoglutarate, alginate, cyclopentanepropionate, 3-phenylpropionate, 3-cyclohexylpropionic acid, 2-naphthylcarboxate, 2-naphthylsulfonate, dihydroxynaphthylate, lauryl sulfate, glycerol phosphate, lauryl sulfate, pectin esters, etc. Alkaline salts include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, and salts of organic bases such as dicyclohexylamine salts and N-methyl-D-glucosamine salts. Furthermore, the basic nitrogen-containing group can be quaternized with reagents such as lower alkyl halides, such as chloro, bromine, and iodides of methyl, ethyl, propyl, and butyl groups; dialkyl sulfates, such as dimethyl, diethyl, dibutyl, and dipentyl sulfates; long-chain halides, such as chloro, bromine, and iodides of decyl, lauryl, myristyl, and stearoyl groups; and aralkyl halides, such as bromides of benzyl and phenethyl groups. Preferred acids for forming acid addition salts include hydrochloric acid, p-toluenesulfonic acid, methanesulfonic acid, maleic acid, malic acid, picric acid, citric acid, and p-aminobenzenesulfonic acid.

[0021] The present invention also relates to pharmaceutical compositions that inhibit tyrosine kinases and serine threonine kinases, the compositions comprising a compound of formula I, a derivative thereof or a pharmaceutically suitable acid addition salt thereof, and a pharmaceutically acceptable carrier.

[0022] "Pharmaceutical acceptable" refers to pharmaceutically acceptable carriers, excipients, prodrugs, etc., which are pharmacologically acceptable and substantially non-toxic to patients who are given the specific compound.

[0023] "Pharmaceutically active metabolites" refer to the metabolites of pharmaceutically acceptable and effective Formula I compounds.

[0024] The term "halogen" as used in this invention includes fluorine, chlorine, bromine, or iodine.

[0025] The compounds of this invention can be administered to patients by various methods, such as oral administration as capsules or tablets, as sterile solutions or suspensions, and in some cases, intravenous injection as solutions. The free base compounds of this invention can be formulated and administered as pharmaceutically suitable acid addition salts.

[0026] The compounds described in this invention, as novel structural types of vascular endothelial growth factor receptor (VEGFR) and epidermal growth factor receptor (EGFR), possess novel structural characteristics and can be used to treat or prevent tumors related to vascular endothelial growth factor receptor-2 (VEGFR-2) kinase and epidermal growth factor receptor (EGFR) kinase, such as lung cancer, colorectal cancer, prostate cancer, and breast cancer. They have good application value and promising development prospects.

[0027] The use of the compounds described in this invention or pharmaceutically acceptable salts thereof, and pharmaceutical compositions comprising the compounds, in the preparation of antitumor drugs.

[0028] The use of the compounds described in this invention or pharmaceutically acceptable salts thereof, and pharmaceutical compositions comprising the compounds, in the preparation of vascular endothelial growth factor receptor kinase and epidermal growth factor receptor kinase inhibitors.

[0029] Preparation route of the compounds of this invention:

[0030]

[0031] Beneficial effects of this invention:

[0032] 1. This invention provides a novel class of antitumor compounds with a novel structure, which can be used to treat or prevent tumors related to vascular endothelial growth factor receptor-2 (VEGFR-2) kinase and epidermal growth factor receptor (EGFR) kinase, such as lung cancer, colorectal cancer, prostate cancer, and breast cancer, and has potential pharmaceutical value.

[0033] 2. Compared with marketed drugs, the compounds of the present invention have stronger anti-tumor cell proliferation activity.

[0034] 3. The compounds and pharmaceutically acceptable salts described in this invention are capable of binding to VEGFR and EGFR, thereby blocking related downstream signaling pathways and inhibiting tumor cell proliferation.

[0035] 4. The compound preparation route of the present invention is simple, easy to synthesize and low in cost. Detailed Implementation

[0036] The present invention is described in detail by way of the following examples. However, it should be understood that the present invention is not limited to the specific examples described below. Example 1: Preparation of 6-(4-methoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide (LB01)

[0037] Step A: Preparation of tert-butyl 2-amino-3-carbamoyl-4,7-dihydrothiopheno[2,3-c]pyridine-6(5H)-carboxylic acid

[0038] Weigh 2.0 g (10 mmol) of 1-Boc-4-piperidinone, 1.3 g (15 mmol) and S (0.5 g, 15 mmol) into a round-bottom flask, add 20 mL of anhydrous ethanol, and add dropwise morpholine (2.6 g, 30 mmol). Heat to reflux and monitor the reaction by TLC until complete. Cool to room temperature, filter to remove S, remove ethanol by vacuum distillation, and separate by column chromatography to give 1.33 g of yellow solid, yield 88.67%. 1 H NMR (600MHz, DMSO-d6) δ6.99 (s, 2H), 6.61 (s, 2H), 4.26 (s, 2H), 3.50 (t, J = 5.7Hz, 2H), 2.67 (q, J = 5.6, 3.9Hz, 2H), 1.41 (s, 9H).

[0039] Step B: Preparation of tert-butyl 3-carbamoyl-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,7-dihydrothiopheno[2,3-c]pyridine-6(5H)carboxylic acid

[0040] 1.4 g (4.7 mmol) of 2-amino-3-carbamoyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester, phenyl[4-(pyrrolidine-1-yl)butyl]carbamate, and 0.6 g (10.7 mmol) of potassium hydroxide were weighed and added to a round-bottom flask. 20 mL of DMSO was added, and the mixture was reacted under argon protection at room temperature. The reaction was monitored by TLC until complete. The reaction solution was added dropwise to 150 mL of water, extracted with DCM, separated, dried over anhydrous sodium sulfate for 2 h, filtered to remove anhydrous sodium sulfate, and distilled under reduced pressure to remove DCM. Column chromatography was used to separate the solid, yielding 1.06 g of a grayish-white solid (yield 48.4%).

[0041] Step C: Preparation of 2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide hydrochloride

[0042] 0.5 g (1.1 mmol) of tert-butyl 3-carbamoyl-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,7-dihydrothieno[2,3-c]pyridine-6(5H)carboxylic acid was weighed and added to a round-bottom flask. 10 mL of hydrochloric acid-ethanol solution was added dropwise. The reaction was carried out at room temperature and monitored by TLC. The reaction was complete. Ethanol was removed by vacuum distillation to give 0.41 g of a yellow solid, yield 85.4%. 1 H NMR(600MHz,DMSO-d6)δ10.76(s,1H),10.59(s,1H),9.58(s,2H),7.80(s,1H),7.40–7.18(m,1H),4.16(s,2H),3.49–3.43(m,2H),3 .33–3.28(m,2H),3.13–3.04(m,4H),3.00–2.90(m,4H),2.01–1.94(m,2H),1.90–1.82(m,2H),1.72–1.63(m,2H),1.50–1.43(m,2H).

[0043] Step D: Preparation of 6-(4-methoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB01)

[0044] Weigh 0.43 g (1 mmol) of 2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide hydrochloride into a round-bottom flask, add 30 mL of methanol, add p-methoxybenzaldehyde (0.7 g, 5 mmol), adjust the pH to 5 with acetic acid and sodium acetate, and stir at room temperature for 2 h under N2 protection. Add sodium cyanoborohydride (0.2 g, 2 mmol), and react at room temperature. The reaction is monitored by TLC and the reaction is complete. Remove methanol by vacuum distillation, add 40 mL of DCM and 20 mL of water, adjust the pH to 9, separate the layers, dry with anhydrous sodium sulfate, filter to remove anhydrous sodium sulfate, and separate by column chromatography to give 0.15 g of pale yellow solid, yield 31.2%; mp: 125.1-126.5℃. 1 H NMR(400MHz,Chloroform-d)δ11.21(s,1H),7.28(d,J=8.5Hz,2H),6.87(d,J=8.5Hz,2H),5.59(s,2H),4.50(s,1H),3.81(s,3 H),3.64(s,2H),3.54(s,2H),3.33–3.23(m,2H),2.85–2.72(m,4H),2.69–2.45(m,6H),1.89–1.81(m,4H),1.68–1.57(m,4H). 13 C NMR (151MHz, DMSO-d6) δ158.75,60.94,55.57,55.45,53.98,51.32,49.93,40.47,27.84,26.13,25.84,23.45.ESI-HRMS m / z:calculated C 25 H 35 N5O3S([M+H)) + ):486.25389,found:486.25259.

[0045] Example 2: Preparation of 6-(4-ethylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB02)

[0046] Following the preparation method of Example 1, 0.21 g of a pale yellow solid was obtained, with a yield of 43.4%; mp: 128.5-130.1℃; 1H NMR (400MHz, Chloroform-d) δ11.18(s,1H),7.49(d,J=7.8Hz,2H),7.16(d,J=7.8Hz,2H),5.72(s,2H),4.52(s,1H),3.66(s,2H),3.55( s,2H),3.28(t,J=6.3Hz,2H),2.80–2.72(m,5H),2.72–2.59(m,6H),1.94–1.82(m,4H),1.73–1.57(m,4H),1.27–1.19(m,4H).ESI-HRMS m / z:calculated C 26 H 37 N5O2S([M+H)) + ):484.27463,found:484.27341.

[0047] Example 3: Preparation of 6-(3-methylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB03)

[0048] Following the preparation method of Example 1, 0.20 g of a white solid was obtained, with a yield of 42.6%; mp: 118.8-120.6℃; 1 HNMR(400MHz,Chloroform-d)δ11.17(s,1H),7.22–7.17(m,2H),7.14–7.05(m,2H),5.75(s,2H),4.52(s,1H),3.66(s,2H),3.5 5(s,2H),3.28(t,J=6.2Hz,2H),2.82–2.72(m,6H),2.70–2.62(m,2H),2.39–2.31(m,5H),1.94–1.82(m,4H),1.73–1.57(m,4H). 13 C NMR(151MHz,DMSO-d6)δ154.13,138.79,137.76,129.88,128.60,128.16,126.35, 112.39,61.56,53.86,52.38,51.48,50.09,27.72,26.15,23.39,21.49.ESI-HRMS m / z:calculatedC 25 H 35 N5O2S([M+H)) + ):470.25898,found:470.25745.

[0049] Example 4: Preparation of 6-(4-chlorobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide (LB04) was carried out according to the preparation method of Example 1, yielding 0.28 g of a pale yellow solid, with a yield of 58.3%; mp: 131.1-133.2℃; 1 H NMR(400MHz,Chloroform-d)δ11.21(s,1H),7.34–7.28(m,4H),5.62(s,2H),4.52(s,1H),3.66(s,2H),3.55(s ,2H),3.32–3.23(m,2H),2.83–2.73(m,4H),2.71–2.52(m,6H),1.91–1.80(m,4H),1.71–1.59(m,4H).ESI-HRMS m / z:calculated C 24 H 32 ClN5O2S([M+H) + ):490.20435,found:490.20337.

[0050] Example 5: Preparation of 6-benzyl-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB05)

[0051] Following the preparation method of Example 1, 0.25 g of a pale yellow solid was obtained, with a yield of 55.6%; mp: 89.5-92.1℃; 1 HNMR(400MHz,Chloroform-d)δ11.19(s,1H),7.63–7.53(m,1H),7.44–7.30(m,4H),5.65(s,2H),4.56(s,1H),3.70(s ,2H),3.56(s,2H),3.29(t,J=6.1Hz,2H),2.82–2.57(m,8H),2.05–1.94(m,2H),1.94–1.81(m,4H),1.72–1.53(m,4H). 13C NMR(151MHz,DMSO-d6)δ167.85,154.13,138.85,129.27,128.72,127.52,112.38,66.38,61.51,60 .77,55.45,53.94,51.44,50.03,48.40,47.17,40.45,27.78,26.12,25.63,23.42,21.46.ESI-HRMS m / z:calculated C 24 H 33 N5O2S([M+H)) + ):456.24333,found:456.24176.

[0052] Example 6: Preparation of 6-(2,3-dimethoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB06)

[0053] Following the preparation method of Example 1, 0.15 g of a pale yellow solid was obtained, with a yield of 29.1%; mp: 91.2-93.4℃; 1 HNMR(400MHz,Chloroform-d)δ11.17(s,1H),7.08–7.01(m,2H),6.87–6.82(m,1H),5.74(s,2H),4.55(s,1H),3.87(s,3H),3.85( s,2H),3.83(s,3H),3.75(s,2H),3.61–3.50(m,4H),2.93–2.70(m,8H),1.96–1.90(m,4H),1.78–1.71(m,2H),1.66–1.60(m,2H). 13 C NMR (151MHz, DMSO-d6) δ152.91,147.57,132.29,124.18,112.00,60.71,56.01,55.37,53.93,52.37,51.58,40.46,27.73,26.32,23.39.ESI-HRMS m / z:calculated C 26 H 37 N5O4S([M+H)) + ):516.26446,found:516.26294.

[0054] Example 7: Preparation of 6-(3,4,5-trimethoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB07)

[0055] Following the preparation method of Example 1, 0.16 g of a pale yellow solid was obtained, with a yield of 29.3%; mp: 84.1-86.4℃; 1 HNMR(400MHz,Chloroform-d)δ11.23(s,1H),6.61(s,2H),5.65(s,2H),4.49(s,1H),3.95–3.75(m,9H),3.63(s,2H), 3.59(s,2H),3.36–3.25(m,2H),2.82–2.74(m,4H),2.73–2.62(m,2H),1.97–1.86(m,4H),1.79–1.54(m,8H).ESI-HRMS m / z:calculated C 27 H 39 N5O5S([M+H)) + ):546.27502,found:546.27386.

[0056] Example 8: Preparation of 6-(4-methylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB08)

[0057] Following the preparation method of Example 1, 0.24 g of a pale yellow solid was obtained, with a yield of 51.1%; mp: 109.6-111.4℃; 1 H NMR(400MHz,Chloroform-d)δ11.18(s,1H),7.24–7.10(m,4H),5.71(s,2H),4.52(s,1H),3.66(s,2H),3.54(s,2H),3.29(t,J= 6.2Hz,2H),2.81–2.73(m,6H),2.67(t,J=7.2Hz,2H),2.35(s,3H),1.93–1.86(m,4H),1.74–1.60(m,4H),1.44(m,2H).ESI-HRMS m / z:calculated C 25 H 35 N5O2S([M+H)) + ):470.25898,found:470.25748.

[0058] Example 9: Preparation of 6-(3-cyanobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB09)

[0059] Following the preparation method of Example 1, 0.27 g of a pale yellow solid was obtained, with a yield of 56.2%; mp: 98.0-100.3℃; 1 HNMR(400MHz,Chloroform-d)δ11.21(s,1H),7.71(s,1H),7.64–7.56(m,2H),7.47–7.41(m,1H),5.61(s,2H),4.58(s,1H),3 .72(s,2H),3.57(s,2H),3.29(t,J=5.7Hz,2H),2.85–2.76(m,4H),2.73–2.53(m,6H),1.91–1.84(m,4H),1.70–1.60(m,4H). 13 C NMR(151MHz,DMSO-d6)δ167.82,154.13,147.08,140.79,134.15,132.57,131.42,130.01,127.43,121 .61,119.38,111.70,60.22,55.47,53.95,51.39,49.88,40.45,27.79,26.01,25.68,23.43.ESI-HRMS m / z:calculated C 25 H 32 N6O2S([M+H)) + ):481.23857,found:481.23737.

[0060] Example 10: Preparation of 6-(4-isobutylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide (LB10)

[0061] Following the preparation method of Example 1, 0.22 g of a pale yellow solid was obtained, with a yield of 43.1%; mp: 148.7-150.2℃. 1H NMR(400MHz,Chloroform-d)δ11.17(s,1H),7.26–7.23(m,1H),7.20–7.16(m, 1H),7.11(d,J=7.7Hz,2H),5.76(s,2H),4.53(s,1H),3.66(s,2H),3.54(s,2H) ,3.34–3.23(m,2H),2.90–2.65(m,8H),2.46(d,J=7.4Hz,2H),2.05–1.79(m,6H ),1.76–1.67(m,2H),1.66–1.59(m,2H),1.46–1.39(m,1H),0.94–0.81(m,6H). 13 C NMR (151MHz, DMSO) δ153.03,139.19,134.97,128.19,128.00,127.17,111.27,60.21,52.81,51.28, 43.68,39.37,39.25,39.11,38.97,38.83,38.69,38.55,38.41,29.07,22.30,21.57,6.54.ESI-HRMS m / z:calculatedC 28 H 41 N5O2S([M+H)) + ):512.30537,found:512.30457.

[0062] Example 11: Preparation of 6-(4-pentylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB11)

[0063] Following the preparation method of Example 1, 0.21 g of a pale yellow solid was obtained, with a yield of 40.4%; mp: 128.9-130.8℃. 1 H NMR(400MHz,Chloroform-d)δ11.19(s,1H),7.29–7.26(m,1H),7.26–7.24(m,1H),7.14(d,J=7.8Hz,2H),5.68(s,2H),3.66(s,2H),3.55(s,2H),3 .33–3.25(m,2H),2.83–2.64(m,8H),2.62–2.56(m,2H),1.95–1.86(m,4H ),1.72–1.55(m,6H),1.39–1.21(m,6H),0.89(t,J=6.9Hz,3H).ESI-HRMS m / z:calculated C29 H 43 N5O2S([M+H)) + ):526.32102,found:526.32001.

[0064] Example 12: Preparation of 6-(3-ethylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB12)

[0065] Following the preparation method of Example 1, 0.23 g of a pale yellow solid was obtained, with a yield of 46.9%; mp: 132.7-134.0℃; 1 H NMR(400MHz,Chloroform-d)δ11.21(s,1H),7.26–7.10(m,4H),5.61(s,2H),4.54(s,1H),3.68(s,2H),3.57(s ,2H),3.29(t,J=6.3Hz,2H),2.87–2.55(m,10H),2.04–1.86(m,6H),1.74–1.59(m,4H),1.24(t,J=7.6Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ153.02,143.06,137.75,127.56,125.85,60.49,54. 40,52.88,50.40,48.98,38.41,27.51,26.70,25.04,22.33,15.09.ESI-HRMS m / z:calculated C 26 H 37 N5O2S([M+H)) + ):484.27463,found:484.27313.

[0066] Example 13: Preparation of 6-(2-methoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB13)

[0067] Following the preparation method of Example 1, 0.23 g of a pale yellow solid was obtained, with a yield of 47.9%; mp: 125.9-128.1℃; 1H NMR(400MHz,Chloroform-d)δ11.20(s,1H),7.40–7.36(m,1H),7.00–6.85(m,3H),5.66(s,2H),4.55(s,1H),3. 83(s,3H),3.74(s,2H),3.62(s,2H),3.29(t,J=6.2Hz,2H),2.91–2.60(m,10H),1.91(s,4H),1.76–1.60(m,4H). 13 C NMR (151MHz, DMSO-d6) δ157.75,154.11,55.93,55.73,55.32,55.07,53.83,52.39,51.57,50.31,40.44,27.74,26.26,25.42,23.40.ESI-HRMS m / z:calculated C 25 H 35 N5O3S([M+H)) + ):486.25389,found:486.25244.

[0068] Example 14: Preparation of 6-(2-chlorobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB14)

[0069] Following the preparation method of Example 1, 0.29 g of a pale yellow solid was obtained, with a yield of 59.3%; mp: 106.0-107.7℃; 1 H NMR(400MHz,Chloroform-d)δ11.20(s,1H),7.53–7.48(m,1H),7.39–7.35(m,1H),7.25–7.18(m,2H),5.67(s,2H),4.66(s,1H),3.82(s,2 H),3.64(s,2H),3.29(t,J=6.1Hz,2H),2.89–2.79(m,4H),2.76–2.67(m,4H),2.63(t,J=7.0Hz,2H),1.92–1.84(m,4H),1.73–1.61(m,4H). 13C NMR (151MHz, DMSO-d6) δ153.04,130.17,128.71,126.47,120.61,57.08,54. 35,52.85,51.28,50.42,49.09,38.41,26.67,25.04,24.52,22.32.ESI-HRMS m / z:calculated C 24 H 32 ClN5O2S([M+H) + ):490.20435,found:490.20300.

[0070] Example 15: Preparation of 6-(3,5-difluorobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide (LB15)

[0071] Following the preparation method of Example 1, 0.33 g of a pale yellow solid was obtained, with a yield of 67.3%; mp: 153.0-154.7℃; 1 H NMR (600MHz, DMSO-d6) δ10.51(s,1H),7.67(s,1H),7.37(s,1H),7.17–7.11(m,1H),7.11–7.05(m,2H),6.74(s,1H),3.68(s,2H),3. 48(s,2H),3.11–3.02(m,2H),2.76(t,J=5.6Hz,2H),2.65(t,J=5.6Hz,2H),2.50–2.38(m,6H),1.75–1.64(m,4H),1.52–1.39(m,4H). 13 C NMR(151MHz,DMSO-d6)δ166.73,153.03,143.06,126.34,120.50,111.30,110 .85,59.05,54.48,52.90,50.30,48.80,38.42,26.74,24.91,22.36.ESI-HRMS m / z:calculatedC 24 H 31 F2N5O2S([M+H) + ):492.22448,found:492.22284.

[0072] Example 16: Preparation of 6-(3,4-dimethoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB16)

[0073] Following the preparation method of Example 1, 0.21 g of a pale yellow solid was obtained, with a yield of 40.7%; mp: 88.4-90.5℃; 1 HNMR(400MHz,Chloroform-d)δ11.21(s,1H),6.95–6.78(m,3H),5.65(s,2H),4.50(s,1H),3.90–3.85(m,6H),3.63(s,2H),3.56( s,2H),3.29(t,J=6.0Hz,2H),2.80–2.69(m,6H),2.63(t,J=6.8Hz,2H),1.92–1.83(m,4H),1.72–1.58(m,4H),1.49–1.37(m,2H). 13 CNMR(151MHz,DMSO-d6)δ166.72,153.01,147.95,60.21,54.79,54.72,54.50,52.90,51.29,50.33,48.86,38.41,26.75,24.76,22.36.ESI-HRMS m / z:calculated C 26 H 37 N5O4S([M+H)) + ):516.26446,found:516.26276.

[0074] Example 17: Preparation of 6-(4-dimethylaminobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide (LB17)

[0075] Following the preparation method of Example 1, 0.25 g of a pale yellow solid was obtained, with a yield of 50.2%; mp: 103.9-105.8℃; 1 H NMR(400MHz,Chloroform-d)δ11.18(s,1H),7.21(d,J=8.6Hz,2H),6.71(d,J=8.7Hz,2H),5.68(s,2H),4.46(s,1H),3.60(s, 2H),3.53(s,2H),3.29(t,J=6.3Hz,2H),2.99–2.90(m,6H),2.81–2.62(m,8H),1.90(s,4H),1.73–1.57(m,4H),1.45(m,2H). 13C NMR (151MHz, DMSO-d6) δ166.73,153.01,148.95,60.14,54.38,52.85,51.29,48.85,39.62,38.41,26.70,25.09,22.34.ESI-HRMS m / z:calculated C 26 H 38 N6O2S([M+H)) + ):499.28553,found:499.28366.

[0076] Example 18: Preparation of 6-(3,5-dimethylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide (LB18)

[0077] Following the preparation method of Example 1, 0.21 g of a pale yellow solid was obtained, with a yield of 43.4%; mp: 158.9-160.8℃.

[0078] Example 19: Preparation of 6-(2-methylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide (LB19)

[0079] Following the preparation method of Example 1, 0.18 g of a pale yellow solid was obtained, with a yield of 38.3%; mp: 103.9-105.8℃; 1 H NMR(400MHz,Chloroform-d)δ11.23(s,1H),7.34–7.29(m,1H),7.20–7.14(m,3H),5.59(s,2H),4.57(s,1H),3.66(s,2H),3. 57(s,2H),3.31(t,J=6.2Hz,2H),2.88–2.67(m,11H),2.38(s,3H),1.77–1.70(m,4H),1.66–1.62(m,2H),1.49–1.43(m,2H). 13 C NMR(151MHz,DMSO-d6)δ167.82,154.12,130.59,130.00,127.54,125.95,112.38 ,59.56,53.92,52.37,51.63,50.14,40.46,27.74,26.17,23.40,19.30.ESI-HRMS m / z:calculatedC 25 H 35 N5O2S([M+H))+ ):470.25898,found:470.25735.

[0080] Example 20: Preparation of 6-(2,5-dichlorobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide (LB20)

[0081] Following the preparation method of Example 1, 0.20 g of a pale yellow solid was obtained, with a yield of 38.2%; mp: 159.3-162.1℃; 1 H NMR(400MHz,Chloroform-d)δ11.18(s,1H),7.55(d,J=2.6Hz,1H),7.28(s,1H),7.18(dd,J=8.5,2.6Hz,1H),5.76(s,2H),4.62(s,1H), 3.77(s,2H),3.65(s,2H),3.31(t,J=6.2Hz,2H),2.93–2.74(m,10H),1.98–1.92(m,4H),1.81–1.73(m,2H),1.67–1.61(m,2H).ESI-HRMS m / z:calculated C 24 H 31 Cl2N5O2S([M+H) + ):524.16538,found:524.16431.

[0082] Table 1. Comparison of Compound Structure Names

[0083]

[0084]

[0085]

[0086]

[0087] Example 21: Inhibitory activity of the test compound on the proliferation of A549, MCF7, HCT116, and PC3 cells

[0088] (1) Experimental materials

[0089] Cell lines: Lung cancer A549, breast cancer MCF7, colorectal cancer HCT116, and prostate cancer PC3 cells were seeded in 96-well plates at densities of 1500, 2200, 800, and 2000 cells / well, respectively, with 100 μL per well, and used after 24 h.

[0090] Target compounds numbered LB01-LB20: dissolved in DMSO and diluted with culture medium to prepare 100 μM, 50 μM, 20 μM, 10 μM, 5 μM, 2.5 μM, and 1.25 μM concentrations. These six different concentrations were stored at -20℃ for later use. The final concentration of DMSO in the culture medium was less than 0.1%.

[0091] Positive control drug: sorafenib.

[0092] MTT: Dissolve in PBS to a concentration of 2 mg / mL and store at -20°C.

[0093] (2) Experimental methods

[0094] The antitumor proliferation activity of A549, MCF7, HCT116, and PC3 cells was evaluated using the MTT assay. A549, HCT116, and PC3 cell lines were cultured on RPMI 1640 medium containing 10% fetal bovine serum (FBS), while the MCF-7 cell line was cultured on DMEM medium containing 10% FBS. When cells reached 80-90% confluence, they were confluent and passaged for no more than 20 passages, then allowed to acclimatize for 24 hours before further treatment. These cells were then placed in 96-well plates and cultured overnight in a humidified environment containing 5% CO2 at 37°C. After 24 hours, different concentrations of the invention's representative compound were added. After another 24 hours of culture, MTT (2 mg / mL) was added, and the cells were cultured for another 4 hours. The culture medium was removed, and the crystals were dissolved in DMSO. The absorbance was measured at 570 nm using a microplate reader (TECANSPECTRA, WetDar, Germany). The cell growth inhibition rate was calculated using the formula: Cell growth inhibition rate = (1 - OD value of drug group / OD value of control group) × 100%. A logarithmic curve was plotted between different concentrations of the test compound and its cell inhibition rate to calculate the corresponding IC50 of the test compound. 50 Values ​​were determined according to the methods described above for representative compounds of the present invention.

[0095] Table 2. Inhibitory activity of the tested compounds against tumor cell proliferation (IC50) 50 (μM)

[0096]

[0097]

[0098] Of the 20 compounds screened, some showed good inhibitory activity against tumor cell proliferation in HCT116, A549, PC3, and MCF7 cell lines. Among them, compound LB11 showed the best IC50 activity against HCT116, A549, PC3, and MCF7 cell lines. 50 The IC50 values ​​for compounds LB10 and LB11 in HCT116 cell lines were lower than those for the positive control drug sorafenib. 50 Its IC50 is lower than that of the positive control drug sorafenib. 50 The IC50 values ​​were 3.038 ± 0.32 and 2.45 ± 0.44 μM, respectively. Compounds LB10, LB11, and LB18, etc., showed IC50 values ​​against A549 cell lines. 50 The IC50 of LB18 is lower than that of the positive control drug sorafenib. 50 It reached 2.73±0.86μM.

[0099] Example 22: In vitro activity of the test compound in inhibiting VEGFR-2 and EGFR

[0100] (1) Experimental materials

[0101] VEGFR-2, Active (0.1 μg / μl), SignalChem; EGFR, Active (0.1 μg / μl), SignalChem; Poly(4:1Glu, Tyr) Peptide (1 mg / ml), SignalChem; Kinase Assay Buffer III, SignalChem; DTT, Aladdin; ADP-Glo TM Kinase Assay, Promega; DMSO, Sinopharm Corporation; Staurosporine, Ceramics Corporation; 384well small volume white plate, Greiner Corporation; 784075 9. Envison, PerkinElmer Corporation.

[0102] (2) Experimental steps

[0103] Thaw the kinases (VEGFR-2 and EGFR enzymes), Poly(4:1 Glu, Tyr) Peptide, kinase assay buffer III (5X buffer), DTT (2M) and ATP (10mM) on ice separately, and keep all the above reagents on ice throughout the experiment.

[0104] Prepare a 1X buffer by mixing 5X buffer with deionized water and add DTT to it. The concentration of DTT in the 1X buffer is 50 μM.

[0105] Add 1 μl / well of the 5X test compound to a white microplate and centrifuge at 1000 rpm for 1 minute. Positive control wells (1% DMSO): 1 μl / well of 1X buffer containing 5% DMSO. Blank control wells: 1 μl / well of 1X buffer containing 5% DMSO. After the kinase is completely thawed, dilute it to 0.75 ng / μl with 1X buffer, and add 2 μl / well to each white microplate, resulting in 1.5 ng of kinase per well. Add 2 μl / well of 1X buffer to each blank control well.

[0106] Prepare a Poly(4:1Glu, Tyr) Peptide / ATP mixture. Add 2 μl / well of the Poly(4:1Glu, Tyr) Peptide / ATP mixture to a white microplate. The concentration of Poly(4:1Glu, Tyr) Peptide is 0.2 mg / ml, and the concentration of ATP is 50 μM. After adding the ATP, centrifuge the microplate at 1000 rpm for 1 minute. After centrifugation, attach a membrane to the microplate, press it firmly, and incubate at 25°C for 1 hour.

[0107] Equilibrate the ADP-Glo™ reagent and Kinase Detection-related reagents required for the Promega kit to room temperature, and mix the Kinase Detection buffer and Kinase Detection Substrate according to the instructions for use.

[0108] After incubation, add 5 μl / well of ADP-Glo™ reagent to a white microplate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 40 minutes. Add 10 μl / well of Kinase Detection mixture to a microplate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 30 minutes. Perform chemiluminescence detection on a plate reader and read the luminescence value (RLU). The inhibition rates of the test compounds against VEGFR-2 and EGFR enzymes are calculated using the following formula:

[0109] Inhibition%=[RLU(Sample)-RLU(1%DMSO)) / (RLU(Blank)-RLU(1%DMSO)]×100%.

[0110] Table 3. Inhibition of VEGFR-2 activity by the test compounds at different concentrations (inhibition rate, %)

[0111]

[0112] Among the tested compounds, all compounds showed varying degrees of inhibitory activity against VEGFR-2 and EGFR enzymes. Compound LB11 exhibited the strongest inhibitory effect on VEGFR-2 enzyme at a concentration of 10 μM, with an inhibition rate of 78.44%, while compound LB10 showed the strongest inhibitory effect on EGFR enzyme at a concentration of 10 μM, with an inhibition rate of 80.47%.

[0113] Formulation Examples

[0114] The following formulation examples are merely illustrative of the scope of protection of the present invention and are not intended to limit it in any way. The active compounds mentioned in the following examples refer to compounds LB01-LB20 obtained in the above examples.

[0115] Example 23: Tablet Formulation

[0116] The active compound is 25-1000 mg, starch is 45 mg, microcrystalline cellulose is 35 mg, polyvinylpyrrolidone (10% aqueous solution) is 4 mL, sodium carboxymethyl cellulose is 4.5 mg, magnesium stearate is 0.5 mg, and talc is 1 mg.

[0117] Example 24: Suspension Formulation

[0118] 0.1-1000 mg of active compound, 50 mg of sodium carboxymethyl cellulose, 1.25 mg of syrup, 0.1 mg of sodium benzoate, 25 mg of flavoring agent, 5 mg of coloring agent, and add pure water to 5 mL.

[0119] Example 25: Aerosol Formulation

[0120] 0.25 mg of active compound, 25-75 mL of ethanol, and 70 mg of propellant 22 (dichlorofluoromethane).

[0121] Example 26: Suppository Formulation

[0122] 250 mg of active compound and 2000 mL of saturated fatty acid glycerides.

[0123] Example 27: Injectable Formulation

[0124] 50 mg of the active compound, 1000 mL of isotonic salt solution.

[0125] Example 28: Ointment Formulation

[0126] 0.025g of micronized active compound, 10g of liquid paraffin, and soft white wax to a total of 100g.

[0127] Example 29: Ointment Formulation

[0128] 0.025g of active compound, 5g of propylene glycol, 5g of sorbitan sesquioleate, 10g of liquid paraffin, and soft white wax to 100g.

[0129] Example 30: Water-in-oil cream formulation

[0130] 0.025g of active compound, 5g of cetyl alcohol, 5g of glyceryl monostearate, 10g of liquid paraffin, 2g of cetyl alcohol polyoxyethylene ether, 0.1g of citric acid, 0.2g of sodium citrate, 35g of propylene glycol, and water to 100g.

[0131] Example 31: Water-in-oil cream formulation

[0132] 0.025g of micronized active compound, 15g of soft white wax, 5g of liquid paraffin, 5g of cetyl alcohol, 2g of Sorbimacrogolstearate (Tween 65 of a specific pharmaceutical excipient grade), 0.5g of dehydrated sorbitan monostearate, 0.2g of sorbic acid, 0.1g of citric acid, 0.2g of sodium citrate, and water to 100g.

[0133] Example 32: Oil-in-water cream formulation

[0134] 0.025g of active compound, 35g of soft white wax, 5g of liquid paraffin, 5g of dehydrated sorbitol sesquioleate, 0.2g of sorbic acid, 0.1g of citric acid, 0.2g of sodium citrate, and water to 100g.

[0135] Example 33: Lotion Formulation

[0136] 0.25 g of active compound, 0.5 mL of isopropanol, 3 mg of carboxyvinyl polymer, 2 mg of NaOH, and water to 1 g.

[0137] Example 34: Formulation of a suspension for injection

[0138] 10 mg of active compound, 7 mg of sodium carboxymethyl cellulose, 7 mg of NaCl, 0.5 mg of polyoxyethylene (20) dehydrated sorbitan monooleate, 8 mg of benzyl alcohol, and sterile water to 1 mL.

[0139] Example 35: Aerosol Formulation for Oral and Nasal Inhalation

[0140] The active compound was 0.1% w / w, sorbitan trioleate was 0.7% w / w, trichlorofluoromethane was 24.8% w / w, dichlorotetrafluoroethane was 24.8% w / w, and dichlorodifluoromethane was 49.6% w / w.

[0141] Example 36: Formulation of atomizing solution

[0142] Add 7 mg of the active compound and 5 mg of propylene glycol to water to a final volume of 10 g.

[0143] Example 37: Powder Formulation for Inhalation

[0144] Fill a capsule with a mixture of the following ingredients: 0.1 mg of micronized active compound and 20 mg of lactose. Inhale the powder using an inhalation device.

[0145] Example 38: Powder Formulation for Inhalation

[0146] The spherical powder is packed into a multi-dose powder inhaler, with each dose containing 0.1 mg of micronized active compound.

[0147] Example 39: Powder Formulation for Inhalation

[0148] The spheroidized powder is loaded into a multi-dose powder inhaler, each dose containing 0.1 mg of micronized active compound and 1 mg of micronized lactose.

[0149] Example 40: Capsule Formulation

[0150] Active compound 1.0 mg, small sugar spheres 321 mg, Aquacoat ECD 30 6.6 mg, acetylated tributyl citrate 0.5 mg, Tween-80 0.1 mg, Eudragit L 100-55 17.5 mg, triethyl citrate 1.8 mg, talc 8.8 mg, defoamer MMS 0.1 mg.

[0151] Example 41: Capsule Formulation of Vaccines

[0152] Active compound 2.0 mg, small sugar globules 305 mg, Aquocoat ECD 30 5.0 mg, acetylsalicylic acid tributyl ester 0.4 mg, Tween-80 0.14 mg, Eudragit NE30 D 12.6 mg, Eudragit S100 12.6 mg, talc 0.16 mg.

[0153] Example 42: Enema Formulation

[0154] Add 2 mg of active compound, 25 mg of sodium carboxymethyl cellulose, 0.5 mg of disodium EDTA, 0.8 mg of methylparaben, 0.2 mg of propylparaben, 7 mg of sodium chloride, 1.8 mg of citric acid, and 0.01 mg of Tween-80 to pure water to 1 mL.

[0155] Example 43: Formulation containing liposomes

[0156] A. Preparation of the drip formulation

[0157] Dipalmitoyl lecithin (45 mg), dimyristoyl lecithin (7 mg), dipalmitoyl phosphatidylglycerol (1 mg), and the active compound (5 mg) were placed in a glass tube. All components were dissolved in chloroform, and most of the solvent was evaporated with N2. Then, the pressure was reduced, thereby forming a lipid film on the surface of the glass tube. An aqueous solution (0.9% NaCl) was added to the lipid, and liposomes were formed at a phase inversion temperature higher than that of the lipid. The resulting suspension contained liposomes ranging in size from very small vesicles to 2 μm.

[0158] B. Preparation of inhalation formulations

[0159] Liposomes were prepared according to Example A, wherein the aqueous solution contained 10% lactose, and the ratio of lactose to lipids was 7:3. The liposome suspension was frozen with dry ice and then freeze-dried to micronize the dried product, resulting in particles with a mass-average aerodynamic diameter (MMAD) of approximately 2 μm.

[0160] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention in any other way. Any person skilled in the art may make changes or modifications to the disclosed technical content to create equivalent embodiments. Any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the protection scope of the present invention.

Claims

1,4,5,6,7-Tetrahydrothieno[2,3-c]pyridine-3-carboxamide compounds or pharmaceutically acceptable salts thereof, characterized in that, Selected from the following compounds: 6-(4-methoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(4-ethylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(3-methylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(4-chlorobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-Benzyl-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(2,3-dimethoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(3,4,5-trimethoxybenzyl)-2-{3-[4-(pyrrolidine-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(4-methylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(3-cyanobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(4-isobutylbenzyl)-2-{3-[4-(pyrrolidine-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(4-pentylbenzyl)-2-{3-[4-(pyrrolidine-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(3-ethylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(2-methoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(2-chlorobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(3,5-difluorobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(3,4-Dimethoxybenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(4-Dimethylaminobenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(3,5-dimethylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(2-methylbenzyl)-2-{3-[4-(pyrrolidone-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide; 6-(3,6-dichlorobenzyl)-2-{3-[4-(pyrrolidine-1-yl)butyl]ureoyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide.

2. A pharmaceutical composition, characterized in that, The active ingredient includes any one of the compounds of claim 1 or pharmaceutically acceptable salts and pharmaceutically acceptable carriers.

3. The use of a 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of vascular endothelial growth factor receptor kinase and epidermal growth factor receptor kinase inhibitors.

4. The use of the pharmaceutical composition of claim 2 in the preparation of vascular endothelial growth factor receptor kinase and epidermal growth factor receptor kinase inhibitors.

5. The use of a 4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating tumors.

6. The use of the pharmaceutical composition of claim 2 in the preparation of a drug for treating tumors.

7. The application according to claim 5 or 6, characterized in that, The tumors mentioned are lung cancer, colorectal cancer, prostate cancer, and breast cancer.

8. The method for preparing the 4,5,6,7-tetrahydrothiopheno[2,3-c]pyridine-3-carboxamide compound according to claim 1, characterized in that, The preparation routes include the following: Wherein, R1 is the corresponding group at the corresponding position of the compound of claim 1.

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