4, 5, 6, 7-tetrahydrothieno [2, 3-c] pyridine-3-formamide compound as well as preparation method and application thereof

Abstract

The invention discloses a 4, 5, 6, 7-tetrahydrothieno [2, 3-c] pyridine-3-formamide compound as well as a preparation method and application thereof, and belongs to the technical field of medicines. Specifically, the structural general formula of the 4, 5, 6, 7-tetrahydrothieno [2, 3-c] pyridine-3-formamide compound or the pharmaceutically acceptable salt thereof is as shown in formula I, and R1 is described in the claims and the specification. The 4, 5, 6, 7-tetrahydrothieno [2, 3-c] pyridine-3-formamide compound provided by the invention or a pharmaceutically acceptable salt thereof can be used as a vascular endothelial growth factor receptor 2 (VEGFR-2) kinase inhibitor and an epidermal growth factor receptor (EGFR) kinase inhibitor, and can be used for preparing a medicine for preventing and treating diseases such as vascular endothelial growth factor receptor 2 (VEGFR-2) kinase inhibitor and epidermal growth factor receptor (EGFR) kinase inhibitor. The compound can be used for preparing drugs for treating or preventing tumor diseases related to vascular endothelial growth factor receptor 2 kinase and epidermal growth factor receptor kinase, and has good application value and development and application prospects. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compounds and a preparation method thereof, and use thereof as vascular endothelial growth factor receptor kinase and epidermal growth factor receptor kinase inhibitors in the preparation of anti-tumor drugs. Background Art

[0002] In today's world, malignant tumors are still the leading cause of death for human beings. With the development of molecular biology technology, the pathogenesis of tumors has been gradually elucidated, and a series of different targets have been gradually discovered. Through rational drug design, small molecules that can target and bind to protein-protein interaction sites can be used to promote or block the biological functions of related proteins, thereby achieving the purpose of disease treatment. This is a hot topic in the field of tumor treatment.

[0003] Imatinib is the first targeted anti-tumor drug, developed and produced by Novartis Pharmaceuticals of Switzerland, which inhibits the growth of cancer cells by selectively targeting specific molecules involved in the growth of cancer cells (Druker, BJ; Talpaz, M.; Resta, DJ; Peng, B.; Buchdunger, E.; Ford, JM; Lydon, NB; Kantarjian, H.; Capdeville, R.; Ohno-Jones, S.; Sawyers, CLE Effectiveness 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 was 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 was developed by Pfizer and marketed under the trade name Inlyta. It was approved in 2012 for the treatment of advanced renal cell carcinoma after failure of one previous systemic treatment. Clinical trials before approval have been reviewed elsewhere. The development of resistance to VEGFR inhibitors in patients with metastatic advanced renal cell carcinoma has led to its combination with PD-1 inhibitors such as avelumab or pembrolizumab. In 2019, it was approved (Grünwald V, Voss MH, Rini BI, et al. Axitinib plus immune checkpoint inhibitor: evidence-and expert-based consensus recommendation for treatment optimisation and management of related adverse events. Br J Cancer, 2020, 123, 898-904). Axitinib's main targets are VEGFR-1, VEGFR-2, VEGFR-3, PDGFRβ, etc. It is a type IIA selective inhibitor of VEGFR with submillimolar potency.

[0005] Cabozantinib was developed by Exelixis and licensed to Bristol-Myers Squibb, marketed as Cometriq (capsules) and Cabometyx (tablets). Cometriq was approved in 2012 for the treatment of patients with progressive 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, the drug was formally approved as a first-line treatment for patients with advanced renal cell carcinoma. In 2019, cabozantinib was approved for the treatment of patients with liver cancer who had previously received sorafenib. The main targets are MET, VEGFR-2, RET, etc. In biochemical assays, cabozantinib showed effective inhibition of VEGFR-2 and MET kinases, IC 50 In the picomolar and nanomolar 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 the present invention is to provide a compound as shown in formula I, its prodrug and pharmaceutically active metabolite and pharmaceutically acceptable salt thereof, and to provide a preparation method thereof and its use in preparing a method for treating and / or preventing tumor diseases associated with vascular endothelial growth factor receptor kinase and epidermal growth factor receptor kinase.

[0007]

[0008] Among them, R 1 Substituted by 1 to 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 compound or a pharmaceutically acceptable salt thereof of the present invention is selected from:

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

[0011] 6-(4-ethylbenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide;

[0012] 6-(3-methylbenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide;

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

[0014] 6-benzyl-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide;

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

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

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

[0018] 6-(4-dimethylaminobenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3,5-dimethylbenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide amine; 6-(2-methylbenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3,6-dichlorobenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-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 the 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compounds of the present invention, their prodrugs and pharmaceutically active metabolites, and any one of their pharmaceutically acceptable salts and a pharmaceutically acceptable carrier or diluent.

[0020] "Pharmaceutically acceptable salts" refers to conventional acid addition salts or base addition salts which retain the biological effectiveness and properties of the compounds of formula I and which are formed with suitable non-toxic organic or inorganic acids or organic or inorganic bases. Acid addition salts include hydrochlorides, hydrobromides, hydroiodides, nitrates, phosphates, sulfates, perchlorates, thiocyanates, bisulfates, persulfates, borates, formates, acetates, propionates, valerates, pivalates, caproates, heptanoates, caprylates, isooctanoates, undecanoates, laurates, palmitates, stearates, oleates, cyclopropionates, oxalates, malonates, succinates, maleates, fumarates, adipates, azelaates, acrylates, crotonates, tiglates, itaconates, sorbates, cinnamates, glycolates, lactates, malates, tartrates, citrates, tartrates, mandelates, benzilates, tropicates, ascorbic acids, gluconates, glucoheptonates, glucarates, mannonates, lactobionates, benzoates, phthalates, terephthalates, furoates, nicotinates, isonicotinates, Salicylate, acetylsalicylate, butyrate, gallate, caffeate, ferulate, picrate, camphorate, camphorsulfonate, methanesulfonate, ethanesulfonate, propanesulfonate, benzenesulfonate, p-toluenesulfonate, p-aminobenzenesulfonate, sulfamate, 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-naphthoate, 2-naphthalenesulfonate, pamoate, lauryl sulfate, glycerophosphate, lauryl sulfate, pectinate, etc. Basic salts include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts of organic bases such as dicyclohexylamine salts, N-methyl-D-glucamine salts, etc., and basic nitrogen-containing groups can be quaternized with such agents as lower alkyl halides such as methyl, ethyl, propyl and butyl chloride, bromide and iodide; dialkyl sulfates such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate and diamyl sulfate; long chain halides such as decyl, lauryl, myristyl and stearyl chloride, bromide and iodide; aralkyl halides such as benzyl and phenethyl bromide, etc. Preferred acids for forming acid addition salts include hydrochloric acid, p-toluenesulfonic acid, methanesulfonic acid, maleic acid, malic acid, picric acid, citric acid, p-aminobenzenesulfonic acid.

[0021] The present invention also relates to a pharmaceutical composition for inhibiting tyrosine kinase and serine threonine kinase, which contains a compound represented by formula I, a derivative thereof or a pharmaceutically suitable acid addition salt thereof, and a pharmaceutically acceptable carrier.

[0022] "Pharmaceutically acceptable" such as pharmaceutically acceptable carriers, excipients, prodrugs, etc., means pharmacologically acceptable and substantially non-toxic to the patient to whom the particular compound is administered.

[0023] "Pharmaceutically active metabolite" refers to a metabolic product of a compound of Formula I that is pharmaceutically acceptable and effective.

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

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

[0026] The compounds of the present invention are vascular endothelial growth factor receptor (VEGFR) and epidermal growth factor receptor (EGFR) of completely new structural types, and have the characteristics of novel structural types, etc. They can be used to treat or prevent tumor diseases 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 have good application value and development and application prospects.

[0027] The invention relates to the use of the compound or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the compound in the preparation of anti-tumor drugs.

[0028] Use of the compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the compound in the preparation of vascular endothelial growth factor receptor kinase and epidermal growth factor receptor kinase inhibitors.

[0029] The preparation route of the compound of the present invention:

[0030]

[0031] Beneficial effects of the present invention:

[0032] 1. The present invention provides a new class of anti-tumor compounds with novel structural types, which can be used to treat or prevent tumor diseases 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 have potential drug value.

[0033] 2. Compared with the drugs already on the market, the compounds described in the present invention have stronger anti-tumor cell proliferation activity.

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

[0035] 4. The compound of the present invention has a simple preparation route, is easy to synthesize and has low cost. DETAILED DESCRIPTION

[0036] The present invention is described in detail with 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-(pyrrolidin-1-yl)butyl]ureido}-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-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylate

[0038] Weigh 1-Boc-4-piperidone (2.0 g, 10 mmol), cyanoacetamide (1.3 g, 15 mmol) and S (0.5 g, 15 mmol) into an eggplant-shaped bottle, add 20 mL of anhydrous ethanol, drop morpholine (2.6 g, 30 mmol), heat to reflux, monitor by TLC, and the reaction is complete. Cool to room temperature, filter to remove S, remove ethanol by vacuum distillation, separate by column chromatography, and obtain 1.33 g of yellow solid, with a yield of 88.67%; 1 H NMR (600 MHz, DMSO-d 6 )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-(pyrrolidin-1-yl)butyl]ureido}-4,7-dihydrothieno[2,3-c]pyridine-6(5H)carboxylate

[0040] Weigh 2-amino-3-carbamoyl-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylic acid tert-butyl ester (1.4g, 4.7mmol), phenyl[4-(pyrrolidin-1-yl)butyl]carbamate (1.86g, 7mmol) and potassium hydroxide (0.6g, 10.7mmol) into an eggplant-shaped bottle, add 20mL DMSO, argon protection, react at room temperature, TLC monitoring, the reaction is complete. The reaction solution is dropped into 150mL water, extracted with DCM, separated, dried over anhydrous sodium sulfate for 2h, filtered to remove anhydrous sodium sulfate, distilled under reduced pressure to remove DCM, separated by column chromatography, and 1.06g of off-white solid is obtained, with a yield of 48.4%.

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

[0042] Weigh tert-butyl 3-carbamoyl-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,7-dihydrothieno[2,3-c]pyridine-6(5H)carboxylate (0.5 g, 1.1 mmol) and add it to an eggplant-shaped bottle. Add 10 mL of hydrochloric acid ethanol solution and react at room temperature. Monitor by TLC and the reaction is complete. Remove ethanol by distillation under reduced pressure to obtain 0.41 g of yellow solid with a yield of 85.4%. 1 H NMR (600 MHz, DMSO-d 6 )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-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide (LB01)

[0044] Weigh 2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide hydrochloride (0.43 g, 1 mmol) and add it to an eggplant-shaped bottle. Add 30 mL of methanol and p-anisaldehyde (0.7 g, 5 mmol). Adjust the pH to 5 with acetic acid and sodium acetate. 2 Protection, stirring at room temperature for 2h, adding sodium cyanoborohydride (0.2g, 2mmol), reacting at room temperature, monitoring by TLC, the reaction was complete. Methanol was removed by distillation under reduced pressure, 40mL DCM and 20mL water were added, pH was adjusted to 9, liquid separation was performed, dried over anhydrous sodium sulfate, anhydrous sodium sulfate was removed by filtration, and separation by column chromatography, 0.15g of light yellow solid was obtained, 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 (151 MHz, DMSO-d 6 )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 N 5 O 3 S([M+H] + ):486.25389,found:486.25259.

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

[0046] Referring to the preparation method of Example 1, 0.21 g of a light yellow solid was obtained with a yield of 43.4%; mp: 128.5-130.1°C; 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 N 5 O 2 S([M+H] + ):484.27463,found:484.27341.

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

[0048] Referring to the preparation method of Example 1, 0.20 g of white solid was obtained with a yield of 42.6%; mp: 118.8-120.6°C; 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 (151 MHz, DMSO-d 6 )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 N 5 O 2 S([M+H]+ ):470.25898,found:470.25745.

[0049] Example 4: Preparation of 6-(4-chlorobenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide (LB04) Referring to the preparation method of Example 1, 0.28 g of a light yellow solid was obtained, with a yield of 58.3%; mp: 131.1-133.2°C; 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 C1N 5 O 2 S([M+H] + ):490.20435,found:490.20337.

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

[0051] Referring to the preparation method of Example 1, 0.25 g of light yellow solid was obtained, with a yield of 55.6%; mp: 89.5-92.1°C; 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). 13 C NMR (151 MHz, DMSO-d 6)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 N 5 O 2 S([M+H] + ):456.24333,found:456.24176.

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

[0053] Referring to the preparation method of Example 1, 0.15 g of a light yellow solid was obtained with a yield of 29.1%; mp: 91.2-93.4°C; 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 (151 MHz, DMSO-d 6 )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 N 5 O 4 S([M+H] + ):516.26446,found:516.26294.

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

[0055] Referring to the preparation method of Example 1, 0.16 g of light yellow solid was obtained, with a yield of 29.3%; mp: 84.1-86.4°C; 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 N 5 O 5 S([M+H] + ):546.27502,found:546.27386.

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

[0057] Referring to the preparation method of Example 1, 0.24 g of light yellow solid was obtained, with a yield of 51.1%; mp: 109.6-111.4°C; 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 N 5 O 2 S([M+H] +):470.25898,found:470.25748.

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

[0059] Referring to the preparation method of Example 1, 0.27 g of light yellow solid was obtained, with a yield of 56.2%; mp: 98.0-100.3°C; 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 (151 MHz, DMSO-d 6 )δ167.82,154.13,147.08,140.79,134.15,132.57,131.42,130.01,127.43,121.61,119.3 8,111.70,60.22,55.47,53.95,51.39,49.88,40.45,27.79,26.01,25.68,23.43.ESI-HRMS m / z:calculated C 25 H 32 N 6 O 2 S([M+H] + ):481.23857,found:481.23737.

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

[0061] Referring to the preparation method of Example 1, 0.22 g of light yellow solid was obtained with a yield of 43.1%; mp: 148.7-150.2°C. 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 N 5 O 2 S([M+H] + ):512.30537,found:512.30457.

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

[0063] Referring to the preparation method of Example 1, 0.21 g of a light yellow solid was obtained with a yield of 40.4%; mp: 128.9-130.8°C. 1H 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 C 29 H 43 N 5 O 2 S([M+H] + ):526.32102,found:526.32001.

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

[0065] Referring to the preparation method of Example 1, 0.23 g of light yellow solid was obtained with a yield of 46.9%; mp: 132.7-134.0°C; 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 (151 MHz, DMSO-d 6 )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 N 5 O 2 S([M+H] +):484.27463,found:484.27313.

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

[0067] Referring to the preparation method of Example 1, 0.23 g of light yellow solid was obtained, with a yield of 47.9%; mp: 125.9-128.1°C; 1 H 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 (151 MHz, DMSO-d 6 )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 N 5 O 3 S([M+H] + ):486.25389,found:486.25244.

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

[0069] Referring to the preparation method of Example 1, 0.29 g of light yellow solid was obtained, with a yield of 59.3%; mp: 106.0-107.7°C; 1H 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). 13 C NMR (151 MHz, DMSO-d 6 )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 C1N 5 O 2 S([M+H] + ):490.20435,found:490.20300.

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

[0071] Referring to the preparation method of Example 1, 0.33 g of light yellow solid was obtained with a yield of 67.3%; mp: 153.0-154.7°C; 1 H NMR (600 MHz, DMSO-d 6 )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 (151 MHz, DMSO-d 6)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 F 2 N 5 O 2 S([M+H] + ):492.22448,found:492.22284.

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

[0073] Referring to the preparation method of Example 1, 0.21 g of light yellow solid was obtained with a yield of 40.7%; mp: 88.4-90.5°C; 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-d 6 )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 N 5 O 4 S([M+H] + ):516.26446,found:516.26276.

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

[0075] Referring to the preparation method of Example 1, 0.25 g of light yellow solid was obtained, with a yield of 50.2%; mp: 103.9-105.8°C; 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). 13 C NMR (151 MHz, DMSO-d 6 )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 N 6 O 2 S([M+H] + ):499.28553,found:499.28366.

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

[0077] Referring to the preparation method of Example 1, 0.21 g of light yellow solid was obtained, with a yield of 43.4%; mp: 158.9-160.8°C

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

[0079] Referring to the preparation method of Example 1, 0.18 g of light yellow solid was obtained with a yield of 38.3%; mp: 103.9-105.8°C;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 (151 MHz, DMSO-d 6 )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 N 5 O 2 S([M+H] + ):470.25898,found:470.25735.

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

[0081] Referring to the preparation method of Example 1, 0.20 g of a light yellow solid was obtained with a yield of 38.2%; mp: 159.3-162.1°C; 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 Cl 2 N 5O 2 S([M+H] + ):524.16538,found:524.16431.

[0082] Table 1 Comparison table of compound structure names

[0083]

[0084]

[0085]

[0086]

[0087] Example 21: Inhibitory activity of the test compound on 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 plated in 96-well plates at a density of 1500, 2200, 800, and 2000 cells / well, respectively, with 100 μL per well, and used after 24 hours.

[0090] Target compounds No. LB01-LB20 were 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, and 6 different concentrations were stored at -20°C for 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 2 mg / mL and store at -20°C.

[0093] (2) Experimental methods

[0094] A549, MCF7, HCT116, and PC3 cells were selected to evaluate the anti-tumor proliferation activity of the test samples using the MTT method. A549, HCT116, and PC3 cell lines were cultured in RPMI 1640 medium containing 10% fetal bovine serum (FBS), and MCF-7 cell lines were cultured in DMEM medium containing 10% fetal bovine serum (FBS). When the cells proliferated to 80-90%, they were merged and then subcultured for no more than 20 generations, and then they were allowed to adapt to the environment for 24 hours before the next step. The cells were placed on a 96-well plate and then incubated in a 5% CO 2Culture overnight in a humid environment and control the temperature at 37°C. After 24 hours, different concentrations of representative compounds of the invention were added. After another 24 hours of culture, MTT (2 mg / mL) was added and cultured for another 4 hours. The culture matrix was removed, the crystals were dissolved in DMSO, and the absorbance was measured at a wavelength of 570 nm using an enzyme reader (TECANSPECTRA, WetDar, Germany). According to the formula: Cell growth inhibition rate = (1- OD value of the drug group / OD value of the control group) × 100%, the cell growth inhibition rate at the corresponding concentration was calculated, and a logarithmic curve was drawn with different concentrations of the test compound and the inhibition rate on the cells to calculate the corresponding IC 50 The representative compounds of the present invention were determined according to the above method.

[0095] Table 2 Inhibitory activity of the tested compounds on tumor cell proliferation (IC 50 , μM)

[0096]

[0097]

[0098] Among the 20 compounds screened above, some compounds showed good tumor cell proliferation inhibition activity against HCT116, A549, PC3, and MCF7 cell lines. Among them, the IC 50 The IC values ​​of compounds LB10 and LB11 on HCT116 cell lines were lower than those of the positive control drug sorafenib. 50 The IC 50 The IC values ​​of compounds LB10, LB11 and LB18 against A549 cell line were 3.038±0.32 and 2.45±0.44μM respectively. 50 The IC of LB18 was lower than that of the positive control drug sorafenib. 50 Reached 2.73±0.86μM.

[0099] Example 22: Test compounds inhibit VEGFR-2 and EGFR activities in vitro

[0100] (1) Experimental Materials

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

[0102] (2) Experimental procedures

[0103] The kinases (VEGFR-2 enzyme and EGFR enzyme) tested in the experiment, Poly (4:1Glu, Tyr) Peptide, kinase assay buffer III (5X buffer), DTT (2M) and ATP (10mM) were thawed on ice respectively, and the above reagents needed to be kept on ice throughout the experiment.

[0104] The 5X buffer was prepared into a 1X buffer with deionized water, and DTT was added thereto. The concentration of DTT in the 1X buffer was 50 μM.

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

[0106] Prepare Poly(4:1Glu, Tyr)Peptide / ATP mixed solution. Take 2μl / well of the mixed solution of Poly(4:1Glu, Tyr)Peptide / ATP into a white microplate. At this time, the concentration of Poly(4:1Glu, Tyr)Peptide is 0.2mg / ml and the concentration of ATP is 50μM. After adding, centrifuge the microplate at 1000 rpm for 1 minute. After centrifugation, stick a film to the microplate, press the film tightly, and incubate at 25°C for 1 hour.

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

[0108] After the incubation, take 5 μl / well ADP-GloTM reagent and add it to the white microplate, centrifuge the microplate at 1000 rpm for 1 minute, and incubate at 25°C for 40 minutes. Take 10 μl / well Kinase Detection mixture and add it to the microplate, centrifuge the microplate at 1000 rpm for 1 minute, and incubate at 25°C for 30 minutes. Perform chemiluminescence detection on the plate reader and read the luminescence value (RLU). The inhibition rate of the test compound on VEGFR-2 enzyme and EGFR enzyme is 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 in vitro activity by the tested compounds at different concentrations (inhibition rate, %)

[0111]

[0112] Among the above-mentioned test compounds, all compounds showed different degrees of inhibitory activity on VEGFR-2 enzyme and EGFR enzyme, among which compound LB11 had the strongest inhibitory effect on VEGFR-2 enzyme at a concentration of 10 μM, with an inhibition rate of 78.44%, and compound LB10 had 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 the present invention, but are not intended to limit the scope of the present invention in any way. The active compounds described in the following examples are compounds LB01-LB20 prepared in the above examples.

[0115] Example 23: Tablet Formulation

[0116] Active compound 25-1000 mg, starch 45 mg, microcrystalline cellulose 35 mg, polyvinyl pyrrolidone (as a 10% aqueous solution) 4 mL, sodium carboxymethyl cellulose 4.5 mg, magnesium stearate 0.5 mg, talc 1 mg.

[0117] Example 24: Suspension Concentrate Formulation

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

[0119] Example 25: Aerosol Formulation

[0120] Active compound 0.25 mg, ethanol 25-75 mL, propellant 22 (chlorodifluoromethane) 70 mg.

[0121] Example 26: Suppository Formulation

[0122] Active compound 250mg, saturated fatty acid glycerides 2000mL.

[0123] Example 27: Injectable Formulation Formulation

[0124] Active compound 50 mg, isotonic saline solution 1000 mL.

[0125] Example 28: Ointment Formulation

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

[0127] Example 29: Ointment Formulation

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

[0129] Example 30: Oil-in-water cream formulation

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

[0131] Example 31: Oil-in-water cream formulation

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

[0133] Example 32: Water-in-oil cream formulation

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

[0135] Example 33: Lotion Formulation

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

[0137] Example 34: Injectable suspension formulation

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

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

[0140] Active compound 0.1% w / w, Sorbitan trioleate 0.7% w / w, Trichlorofluoromethane 24.8% w / w, Dichlorotetrafluoroethane 24.8% w / w, Dichlorodifluoromethane 49.6% w / w.

[0141] Example 36: Nebulized solution formulation

[0142] Active compound 7 mg, propylene glycol 5 mg, add water to 10 g.

[0143] Example 37: Powder Formulation for Inhalation

[0144] Fill a clear gelatin capsule with the mixture of the following ingredients, 0.1 mg of micronized active compound, 20 mg of lactose, and inhale the powder with the aid of an inhalation device.

[0145] Example 38: Powder Formulation for Inhalation

[0146] The spheronized powder is filled into a multiple-dose powder inhaler, each dose containing 0.1 mg of the micronized active compound.

[0147] Example 39: Powder Formulation for Inhalation

[0148] The spheronized powder is filled into a multiple-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, acetyl 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, defoaming agent MMS 0.1 mg.

[0151] Example 41: Capsule Seedling Formulation

[0152] Active compound 2.0 mg, small sugar spheres 305 mg, Aquocoat ECD 30 5.0 mg, acetyl tributyl citrate 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] Active compound 2 mg, sodium carboxymethyl cellulose 25 mg, disodium edetate 0.5 mg, methyl parahydroxybenzoate 0.8 mg, propyl parahydroxybenzoate 0.2 mg, sodium chloride 7 mg, citric acid 1.8 mg, Tween-80 0.01 mg, add purified water to 1 mL.

[0155] Example 43: Formulation containing liposomes

[0156] A. Preparation of Instillation Formulation

[0157] Dipalmitoylphosphatidylcholine (45 mg), dimyristoylphosphatidylcholine (7 mg), dipalmitoylphosphatidylglycerol (1 mg) and active compound (5 mg) were placed in a glass tube, and all components were dissolved in chloroform and heated with N 2 Most of the solvent was evaporated and 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 to form liposomes at a temperature higher than the phase transition temperature of the lipid. The resulting suspension contained liposomes ranging in size from extremely small vesicles to 2 μm.

[0158] B. Preparation of Formulations for Inhalation

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

[0160] The above is only a preferred embodiment of the present invention, and does not limit the present invention in other forms. Any technician familiar with the profession may use the above disclosed technical content to change or modify it into an equivalent embodiment with equivalent changes. Any simple modification, equivalent change and modification made to the above embodiment according to the technical essence of the present invention without departing from the technical solution of the present invention still belongs to the protection scope of the technical solution of the present invention.

Claims

1. 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compounds or pharmaceutically acceptable salts thereof, It is characterized in that The structural formula is shown in Formula I: Among them, R 1 Substituted by 1 to 3 substituents selected from hydrogen, methyl, methoxy, ethyl, halogen, cyano, isobutyl, n-pentyl, and dimethylamino.

2. The 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compound or a pharmaceutically acceptable salt thereof according to claim 1, It is characterized in that Selected from the following compounds: 6-(4-methoxybenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(4-ethylbenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3-methylbenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(4-chlorobenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-benzyl-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(2,3-dimethoxybenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3,4,5-trimethoxybenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(4-methylbenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3-cyanobenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(4-isobutylbenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(4-pentylbenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3-ethylbenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(2-methoxybenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(2-chlorobenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3,5-difluorobenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3,4-dimethoxybenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(4-dimethylaminobenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3,5-dimethylbenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(2-methylbenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide; 6-(3,6-Dichlorobenzyl)-2-{3-[4-(pyrrolidin-1-yl)butyl]ureido}-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide.

3. A pharmaceutical composition, It is characterized in that A compound comprising any one of the compounds or pharmaceutically acceptable salts of claim 1 or 2 as an active ingredient and a pharmaceutically acceptable carrier or diluent.

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

5. Use of the pharmaceutical composition according to claim 3 in the preparation of vascular endothelial growth factor receptor kinase and epidermal growth factor receptor kinase inhibitors.

6. Use of the 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 2 in the preparation of a drug for treating tumors.

7. Use of the pharmaceutical composition according to claim 3 in the preparation of drugs for treating tumors.

8. The use according to claim 6 or 7, It is characterized in that The tumors include lung cancer, colorectal cancer, prostate cancer, and breast cancer.

9. A method for preparing the 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compounds according to any one of claims 1 to 2, It is characterized in that The 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxamide compounds include the following preparation routes:

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