Thiophene[3,2-d]pyrimidine compounds containing piperidine amide structure and their applications

Abstract

This invention relates to thieno[3,2-d]pyrimidine compounds containing a piperidine amide structure and their applications, belonging to the field of pharmaceutical technology. The thieno[3,2-d]pyrimidine compounds containing a piperidine amide structure have the structure shown in general formula (I) and can be pharmaceutically processed by acidification to form salts. Pharmacological activity screening results show that they have good antitumor activity against human lung adenocarcinoma cell line A549 and human liver cancer cell line HepG2, demonstrating promising prospects for antitumor drug development and application.

Description

Technical Field

[0001] This invention pertains to the pharmaceutical field, specifically relating to thieno[3,2-d]pyrimidine derivatives containing a piperidine amide structure as shown in general formula (I) and their pharmaceutically acceptable salts, methods for their preparation, and pharmaceutical compositions containing said compounds. This invention also relates to the use of these compounds and their pharmaceutically acceptable salts in the preparation of medicaments for treating and / or preventing cancer. Background Technology

[0002] Cancer (also known as malignant tumors) has always been one of the most prevalent diseases threatening human health. In 2020, there were 19.3 million new cancer cases and approximately 10 million cancer deaths worldwide. Both the incidence and mortality rates of cancer are rapidly increasing. Therefore, cancer has become one of the major diseases threatening human life and health. Currently, anti-tumor treatments mainly include surgery, chemotherapy, and immunotherapy. Among these, chemotherapy is currently the primary treatment for malignant tumors; however, in clinical application, existing anti-tumor drugs often exhibit drawbacks such as easy development of drug resistance and significant toxic side effects. Therefore, developing novel, highly effective, and low-toxicity anti-tumor drugs is of significant scientific importance.

[0003] Thiophene-[3,2-d]pyrimidine compounds are an important class of heterocyclic compounds with antitumor, antibacterial, antiviral, and anti-inflammatory biological activities. Among them, thieno[3,2-d]pyrimidine, as the dominant skeletal structure, is widely used in medicinal chemistry research, especially in the research and development of antitumor drugs. In recent years, modification of the thieno[3,2-d]pyrimidine core has become a hot area in new drug development. Various small molecule thieno[3,2-d]pyrimidine compounds with antitumor activity have been reported, such as the epidermal growth factor receptor (EGFR) inhibitor Olmutinib, the p21 protein-activated kinase 4 (PAK4) inhibitor PF-03758309, the aurora kinase inhibitor SNS-314, the phosphatidylinositol kinase 3 (PI3K) inhibitor Apitolisib, the PI3K and histone deacetylase (HDAC) dual inhibitor Fimepinostat, and vascular endothelial growth factor receptor (VEGFR) inhibitors. However, current research on thieno[3,2-d]pyrimidine antitumor drugs faces several challenges: significant toxic side effects in clinical application, unsatisfactory clinical therapeutic effects and pharmacokinetic parameters, and low oral bioavailability. Therefore, developing novel, safe, and effective c-Met kinase inhibitors remains a key area of ​​research for antitumor drugs both domestically and internationally. Summary of the Invention

[0004] The purpose of this invention is to design and synthesize a series of novel thieno[3,2-d]pyrimidine derivatives containing piperidine amide structures. In vitro activity screening showed that these compounds possess antitumor activity and can be used in the development of antitumor drugs. In the prior art, there are no reports on the thieno[3,2-d]pyrimidine derivatives containing piperidine amide structures described in this application or their application in antitumor drug research.

[0005] To achieve the objectives of this invention, this invention provides a thieno[3,2-d]pyrimidine compound having a piperidine amide structure as shown in general formula (I) and its pharmaceutically acceptable salt.

[0006]

[0007] in:

[0008] R is selected from adamantyl alkyl group, C6-C 10 Yuanfangyi or C6-C 10 Heteroaryl, wherein the aryl group is replaced by any 1 to 3 identical or different R1 groups.

[0009] R1 is selected from hydrogen, halogen, C1-C6 alkyl, or C1-C6 alkoxy.

[0010] Furthermore, the aforementioned thieno[3,2-d]pyrimidine compounds containing piperidine amide structures and their pharmaceutically acceptable salts have two types of substructures shown in general formula (Ia) or (Ib).

[0011]

[0012] in:

[0013] R is selected from adamantyl, phenyl, or pyridyl, wherein the phenyl and pyridyl groups can be replaced by any 1 to 3 identical or different R1 groups.

[0014] R1 is selected from hydrogen, fluorine, chlorine, methyl, or methoxy.

[0015] Furthermore, thieno[3,2-d]pyrimidine compounds of general formula (Ia) or (Ib) containing a piperidineamide structure and their pharmaceutically acceptable salts are selected from the following compounds:

[0016] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-phenylpiperidin-4-carboxamide

[0017] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-fluorophenyl)piperidin-4-carboxamide

[0018] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(2-fluorophenyl)piperidin-4-carboxamide

[0019] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-chlorophenyl)piperidin-4-carboxamide

[0020] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-methylphenyl)piperidin-4-carboxamide

[0021] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-methoxyphenyl)piperidine-4-carboxamide

[0022] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(3,4-dimethoxyphenyl)piperidine-4-carboxamide

[0023] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(1-adamantyl)piperidin-4-carboxamide

[0024] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(2,4-difluorophenyl)piperidin-4-carboxamide

[0025] 1-(2-chlorothiopheno[3,2-d]pyrimidin-4-yl)-N-phenylpiperidin-3-carboxamide

[0026] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-fluorophenyl)piperidine-3-carboxamide

[0027] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(2-fluorophenyl)piperidine-3-carboxamide

[0028] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-chlorophenyl)piperidine-3-carboxamide

[0029] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-methylphenyl)piperidin-3-carboxamide

[0030] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-methoxyphenyl)piperidine-3-carboxamide

[0031] 1-(2-chlorothiopheno[3,2-d]pyrimidin-4-yl)-N-(3,4-dimethoxyphenyl)piperidine-3-carboxamide

[0032] 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(1-adamantyl)piperidine-3-carboxamide

[0033] A pharmaceutical composition comprising the above-described thieno[3,2-d]pyrimidine compounds containing a piperidine amide structure and their pharmaceutically acceptable salts as active ingredients and pharmaceutically acceptable excipients.

[0034] The use of the above-mentioned thieno[3,2-d]pyrimidine compounds containing the piperidine amide structure and their pharmaceutically acceptable salts or pharmaceutical compositions in the preparation of drugs for treating and / or preventing cancer.

[0035] Furthermore, the cancer is selected from lung cancer and liver cancer.

[0036] Furthermore, according to some common methods in the field to which this invention pertains, the thieno[3,2-d]pyrimidine derivatives containing a piperidine amide structure represented by general formula (I) of this invention can react with acids to form pharmaceutically acceptable salts. Pharmaceutically acceptable addition salts include addition salts of inorganic and organic acids, with salts reacting with the following acids being particularly preferred: hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid, etc.

[0037] The beneficial effects of this invention are:

[0038] This invention demonstrates, through in vitro inhibition assays of human lung adenocarcinoma cells A549 and human liver cancer cells HepG2, that the compounds of this invention have significant inhibitory effects on human lung adenocarcinoma cells and human liver cancer cells, and are particularly useful for the preparation of drugs for the treatment and / or prevention of lung cancer and liver cancer. Detailed Implementation

[0039] The examples and preparation methods provided below further illustrate and demonstrate the compounds of the present invention and their preparation methods. It should be understood that the scope of the following examples and preparation methods does not limit the scope of the invention in any way. The following examples are intended to illustrate, rather than limit, the scope of the invention. The proton NMR spectra of the compounds were determined using a Bruker ARX-300, ARX-400, or ARX-600, and the mass spectra were determined using an Agilent 6460QQQ; the infrared spectra of the compounds were determined using a Perkin-Elmer Spectrumone FT-IR spectrometer (potassium bromide pellet method), and all reagents used were analytically pure or chemically pure.

[0040] The following synthetic routes describe the preparation of the general formula (I) derivatives of the present invention. All starting materials are prepared by the synthetic routes described below, by methods well known to those skilled in the art of organic chemistry, or are commercially available. All final derivatives of the present invention are prepared by the synthetic routes described below or by similar methods well known to those skilled in the art of organic chemistry. All variable factors used in the following synthetic routes are as defined below or as defined in the claims. The synthetic routes are as follows:

[0041]

[0042] General preparation method

[0043] Synthesis of step A2,4-dihydroxythiopheno[3,2-d]pyrimidine (a)

[0044] 76.40 g (1.27 mol) of urea and 40.00 g (254.47 mmol) of methyl 3-aminothiophene-2-carboxylic acid were thoroughly mixed and heated to 190 °C for 4 h to melt. The reaction mixture was cooled to 120 °C and added to 250 mL of 20% NaOH solution while hot, and stirred at room temperature for 1 h. Insoluble matter was removed by filtration, and the pH of the filtrate was adjusted to 3.0 with 2.0 mol / L hydrochloric acid solution. The mixture was stirred at room temperature for 0.5 h, and the precipitated solid was filtered. The filter cake was washed with water until neutral and dried under vacuum at 50 °C for 24 h to obtain 34.70 g of grayish-white solid 2,4-dihydroxythiopheno[3,2-d]pyrimidine(a), yield 81.1%, mp 102–104 °C (literature value: 101–103 °C); MS (ESI), m / Z: C6H5N2O2S[M+H] + Theoretical value 169.01, measured value 169.10.

[0045] Step B: Synthesis of 2,4-dichlorothiopheno[3,2-d]pyrimidine (b)

[0046] 35.00 g (208.12 mmol) of 2,4-dihydroxythiopheno[3,2-d]pyrimidine (a) was weighed and added in portions to 280 mL of stirred phosphorus oxychloride. The mixture was refluxed overnight. Excess phosphorus oxychloride was removed by depressurization evaporation. The reaction mixture was slowly added to ice water under stirring and stirred at room temperature for 0.5 h. The precipitated solid was filtered, and the filter cake was washed with water until neutral. After drying under reduced pressure at 50 °C for 24 h, 31.20 g of grayish-white solid 2,4-dichlorothiopheno[3,2-d]pyrimidine (b) was obtained, with a yield of 73.1% and mp 138–140 °C (literature value: 138.8–139.3 °C). MS (ESI), m / Z: theoretical value C6H3Cl2N2S[M+H] + 204.94, measured value 205.10.

[0047] Synthesis of step C1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)piperidine-4-carboxylate methyl ester (c) and 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)piperidine-3-carboxylate methyl ester (e)

[0048] 6.16 g (30.04 mmol) of 2,4-dichlorothiopheno[3,2-d]pyrimidine (b), 12.46 g (90.12 mmol) of potassium carbonate, and 5.60 g (39.11 mmol) of methyl 4-piperidinecarboxylate or methyl 3-piperidinecarboxylate were added to 60 mL of acetonitrile and refluxed for 10 h. Most of the solvent was removed by evaporation under reduced pressure. The remaining mixture was extracted with 100 mL of water and 80 mL of dichloromethane. The dichloromethane layer was separated, and the aqueous layer was further extracted with dichloromethane (2 × 60 mL). The resulting dichloromethane layers were combined, washed with water until neutral, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by evaporation under reduced pressure to obtain the crude product. The crude product was separated by silica gel column chromatography to obtain a white solid. That is: methyl 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)piperidine-4-carboxylate (c), 6.15 g, yield 65.7%; HRMS (ESI), m / Z: theoretical value C 13 H 15 ClN3O2S[M+H] + 312.0574, measured value 312.0578. Methyl 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)piperidine-3-carboxylate (e), 6.88 g, yield 73.5%; HRMS (ESI), m / Z: theoretical value C 13 H 15 ClN3O2S[M+H] + 312.0574, measured value 312.0577.

[0049] Synthesis of step D1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)piperidine-4-carboxylic acid (d) and 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)piperidine-3-carboxylic acid (f)

[0050] 5.00 g (16.04 mmol) of intermediate (c) or intermediate (e), 45 mL of methanol, and 15 mL of water were added to a reaction flask. 1.92 g (48.11 mmol) of sodium hydroxide was added under stirring, and the reaction was carried out at room temperature for 18 h. After the reaction was complete, the methanol was removed by vacuum concentration. 80 mL of water was added to the reaction flask, and the mixture was extracted twice with 80 mL of ethyl acetate. The ethyl acetate phase was discarded, and the pH of the aqueous phase was adjusted to 3–4 with concentrated hydrochloric acid. A large amount of white solid precipitated from the solution. The solid was filtered, and the filter cake was washed with water until neutral. After drying under reduced pressure at 50 °C for 24 h, 4.27 g of 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)piperidine-4-carboxylic acid (d) was obtained, with a yield of 89.4%. HRMS (ESI), m / Z: theoretical value C 12 H 13 ClN3O2S[M+H] + 298.0417, measured value 298.0418; 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)piperidine-3-carboxylic acid (f), 4.36 g, yield 91.3%; HRMS (ESI), m / Z: theoretical value C 12 H 13 ClN3O2S[M+H] + 298.0417, measured value 298.0420.

[0051] Step E: General Method for Preparing Compounds of Formulas Ia to Ib

[0052] 0.89 g (3.00 mmol) of intermediate (d) or intermediate (f), 3.60 mmol of substituted aniline or adamantane, 0.40 g (6.00 mmol) of triethylamine (TEA), and 1.60 g (4.20 mmol) of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) were dissolved in 10 mL of N,N-dimethylformamide (DMF) and reacted at room temperature for 12 h with stirring. After the reaction was complete, the reaction solution was poured into 50 mL of 20% sodium carbonate solution, and extracted three times with 30 mL of dichloromethane each time. The dichloromethane phases were combined and washed three times with 30 mL of 20% sodium carbonate solution each time. The dichloromethane phase was then washed three times with 50 mL of saturated brine each time. The mixture was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum evaporation to obtain the crude product. The crude product was separated by silica gel column chromatography to obtain white solid 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-substituted piperidine amides (Ia-1~9 and Ib1~8).

[0053] Example 1: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-phenylpiperidin-4-carboxamide (Ia-1): white solid powder, yield 71.4%; 1H NMR (600MHz, DMSO-d6), δ: 9.97 (s, 1H), 8.30 (d, J = 5.5Hz, 1H), 7.60 (d, J = 7.7Hz, 2H), 7.41 (d, J = 5.5Hz, 1H), 7.29 (t, J = 7.9Hz, 2H) ,7.03(t,J=7.4Hz,1H),4.67(d,J=13.4Hz,2H),3.44–3.33(m,2H),2.85–2.70(m,1H),2.01(d,J=10.7Hz,2H),1.80–1.65(m,2H); 13 C NMR (150MHz, DMSO-d6), δ: 173.03, 163.15, 158.20, 156.57, 139.69, 135.67, 129.14, 124.33, 123.61, 119.66, 112.89, 45.95, 42.75, 28.73; HRMS (ESI), m / Z: theoretical value C 18 H 18 ClN4OS[M+H] + 373.0890, measured value 373.0891.

[0054] Example 2: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-fluorophenyl)piperidine-4-carboxamide (Ia-2): white solid powder, yield 69.3%; 1 H NMR (600MHz, DMSO-d6), δ: 10.04 (s, 1H), 8.30 (d, J = 5.5Hz, 1H), 7.71–7.54 (m, 2H), 7.41 (d, J = 5.5Hz, 1H), 7.13 (t, J = 8. 9Hz,2H),4.67(d,J=13.4Hz,2H),3.37(t,J=12.1Hz,2H),2.83–2.70(m,1H),2.00(d,J=10.7Hz,2H),1.82–1.64(m,2H); 13 C NMR (150MHz, DMSO-d6), δ: 172.91, 163.15, 158.37 (d, J = 239.4Hz), 158.19, 156.56, 136.08, 135.67, 124.33, 121.42, 121.36, 115.76, 115.61, 112.89, 45.93, 42.68, 28.70; HRMS (ESI), m / Z: theoretical value C 18 H 17 ClFN4OS[M+H] + 391.0796, measured value 391.0796.

[0055] Example 3: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(2-fluorophenyl)piperidine-4-carboxamide (Ia-3): white solid powder, yield 65.9%; 1 ¹H NMR (600MHz, DMSO-d⁶), δ: 9.78(s, 1H), 8.30(d, J = 5.5Hz, 1H), 7.84(t, J = 8.7Hz, 1H), 7.41(d, J = 5.5Hz, 1H), 7.31–7.21(m, 1H), 7.20–7.06(m, 2H), 4.66(d, J = 13.4Hz, 2H), 3.38(d, J = 13.8Hz, 2H), 3.04–2.86(m, 1H), 2.02(d, J = 11.1Hz, 2H), 1.86–1.60(m, 2H); HRMS (ESI), m / Z: theoretical value C 18 H 17 ClFN4OS[M+H] + 391.0796, measured value 391.0798.

[0056] Example 4: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-chlorophenyl)piperidine-4-carboxamide (Ia-4): white solid powder, yield 78.4%; 1 H NMR (600MHz, DMSO-d6), δ: 10.18 (s, 1H), 8.27 (d, J = 5.5Hz, 1H), 7.63 (d, J = 8.9Hz, 2H), 7.41 (d, J = 5.5Hz, 1H), 7.36 (d, J = 8 .8Hz,2H),4.68(d,J=13.5Hz,2H),3.35(t,J=12.0Hz,2H),2.87–2.68(m,1H),2.01(d,J=10.9Hz,2H),1.79–1.60(m,2H); 13 C NMR (150MHz, DMSO-d6), δ: 173.36, 163.00, 158.12, 156.52, 138.42, 135.65, 129.05, 127.33, 124.24, 121.34, 112.86, 45.90, 42.72, 28.64; HRMS (ESI), m / Z: theoretical value C 18 H 17 Cl2N4OS[M+H] + 407.0500, measured value 407.0501.

[0057] Example 5: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-methylphenyl)piperidine-4-carboxamide (Ia-5): white solid powder, yield 73.3%; 1 H NMR (600MHz, DMSO-d6), δ: 9.88 (s, 1H), 8.30 (d, J = 5.5Hz, 1H), 7.48 (d, J = 8.4Hz, 2H), 7.41 (d, J = 5.5Hz, 1H), 7.09 (d, J = 8.2Hz, 2 H),4.66(d,J=13.4Hz,2H),3.37(t,J=10.8Hz,2H),2.84–2.67(m,1H),2.24(s,3H),1.99(d,J=10.8Hz,2H),1.82–1.62(m,2H); 13 C NMR (150MHz, DMSO-d6), δ: 172.79, 163.15, 158.20, 156.57, 137.18, 135.68, 132.48, 129.50, 124.34, 119.69, 112.89, 45.96, 42.71, 28.75, 20.90; HRMS (ESI), m / Z: theoretical value C 19 H 20 ClN4OS[M+H] + 387.1046, measured value 387.1047.

[0058] Example 6: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-methoxyphenyl)piperidine-4-carboxamide (Ia-6): white solid powder, yield 78.9%; 1H NMR (600MHz, DMSO-d6), δ: 9.83 (s, 1H), 8.30 (d, J = 5.5Hz, 1H), 7.51 (d, J = 9.0Hz, 2H), 7.41 (d, J = 5.5Hz, 1H), 6.87 (d, J = 9.0H 13C NMR (150MHz, DMSO-d6), δ: 172.51, 163.14, 158.19, 156.57, 155.60, 135.66, 132.83, 124.33, 121.21, 114.25, 112.88, 55.61, 45.98, 42.64, 28.78; HRMS (ESI), m / Z: theoretical value C19H20ClN4O2S[M+H]+403.0995, measured value 403.0997.

[0059] Example 7: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(3,4-dimethoxyphenyl)piperidine-4-carboxamide (Ia-7): white solid powder, yield 76.1%; 1 H NMR (600MHz, DMSO-d6), δ: 9.83 (s, 1H), 8.30 (d, J = 5.5Hz, 1H), 7.41 (d, J = 5.5Hz, 1H), 7.33 (d, J = 2.2Hz, 1H), 7.10 (dd, J = 8.7, 2.2Hz, 1H), 6.87 (d, J = 8.7Hz,1H),4.66(d,J=13.4Hz,2H),3.71(s,3H),3.70(s,3H),3.36(t,J=1 2.2Hz,2H),2.80–2.67(m,1H),1.99(d,J=10.8Hz,2H),1.84–1.63(m,2H); 13 C NMR (150MHz, DMSO-d6), δ: 172.57, 163.15, 158.20, 156.57, 148.96, 145.20, 135.68, 133.34, 124.33, 112.88, 112.53, 111.54, 104.86, 56.20, 55.79, 45.96, 42.73, 28.76; HRMS (ESI), m / Z: theoretical value C20 H 22 ClN4O3S[M+H] + 433.1101, measured value 433.1104.

[0060] Example 8: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(1-adamantyl)piperidine-4-carboxamide (Ia-8): white solid powder, yield 81.0%; 1 H NMR (600MHz, DMSO-d6), δ: 8.28 (d, J=5.5Hz, 1H), 7.39 (d, J=5.5Hz, 1H), 7.30 (s, 1H), 4.60 (d, J= 13.4Hz,2H),3.25(t,J=11.8Hz,2H),2.50–2.43(m,1H),2.13–1.73(m,11H),1.69–1.43(m,8H); 13 C NMR (150MHz, DMSO-d6), δ: 173.48, 163.10, 158.16, 156.56, 135.61, 124.31, 112.84, 50.97, 46.06, 42.22, 41.46, 36.52, 29.27, 28.94; HRMS (ESI), m / Z: theoretical value C 22 H 28 ClN4OS[M+H] + 431.1672, measured value 431.1675.

[0061] Example 9: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(2,4-difluorophenyl)piperidine-4-carboxamide (Ia-9): white solid powder, yield 70.2%; 1 H NMR(600MHz, DMSO-d6), δ:9.79(s,1H),8.29(d,J=5.5Hz,1H),7.76(dd,J=15.3,8.9Hz,1H),7.41(d,J=5.5Hz,1H),7.36–7.23(m,1 H),7.05(t,J=7.9Hz,1H),4.78–4.52(m,2H),3.34(br,J=4.1Hz,2H),2.99–2.80(m,1H),2.01(d,J=10.9Hz,2H),1.83–1.61(m,2H); 13C NMR (150MHz, DMSO-d6), δ: 173.49, 163.13, 157.37 (d, J = 247.8Hz), 135.67, 126.58, 124.32, 123.05, 112.89, 111.60, 104.59, 45.93, 41.96, 28.73; HRMS (ESI), m / Z: theoretical value C 18 H 16 ClF2N4OS[M+H] + 409.0701, measured value 409.0702.

[0062] Example 10: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-phenylpiperidin-3-carboxamide (Ib-1): white solid powder, yield 72.8%; 1 H NMR (600MHz, DMSO-d6), δ: 10.03 (s, 1H), 8.28 (d, J = 5.5Hz, 1H), 7.59 (d, J = 7.7Hz, 2 H),7.40(d,J=5.5Hz,1H),7.30(t,J=7.9Hz,2H),7.05(t,J=7.4Hz,1H),4.68(d,J=1 3.5Hz,1H),4.55(d,J=13.2Hz,1H),3.59–3.43(m,1H),3.38(m,1H),2.78–2.64(m,1 H),2.09(d,J=9.9Hz,1H),2.00–1.89(m,1H),1.88–1.74(m,1H),1.68–1.49(m,1H); 13 C NMR (150MHz, DMSO-d6), δ: 171.86, 163.18, 158.24, 156.53, 139.42, 135.74, 129.19, 124.32, 123.81, 119.75, 112.92, 48.57, 46.86, 43.51, 28.24, 24.78; HRMS (ESI), m / Z: theoretical value C 18 H 18 ClN4OS[M+H] + 373.0890, measured value 373.0893.

[0063] Example 11: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-fluorophenyl)piperidine-3-carboxamide (Ib-2): white solid powder, yield 71.1%; 1H NMR (600MHz, DMSO-d6), δ: 10.12 (s, 1H), 8.29 (d, J = 5.5Hz, 1H), 7.75–7.56 (m ,2H),7.41(d,J=5.5Hz,1H),7.15(t,J=8.9Hz,2H),4.68(d,J=13.2Hz,1H),4. 54(d,J=13.3Hz,1H),3.57–3.44(m,1H),3.42–3.36(m,1H),2.72–2.60(m,1H) ,2.17–2.02(m,1H),1.98–1.88(m,1H),1.88–1.77(m,1H),1.68–1.51(m,1H); 13 C NMR (150MHz, DMSO-d6), δ: 171.74, 163.16, 158.46 (d, J = 239.8Hz), 158.20, 156.51, 135.81, 135.80, 135.74, 124.31, 121.47 (d, J = 7.8Hz), 115.76 (d, J = 22.2Hz), 112.89, 48.50, 46.85, 43.41, 28.20, 24.76; HRMS (ESI), m / Z: theoretical value C 18 H 17 ClFN4OS[M+H] + 391.0796, measured value 391.0797.

[0064] Example 12: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(2-fluorophenyl)piperidine-3-carboxamide (Ib-3): white solid powder, yield 68.6%; 1 H NMR (600MHz, DMSO-d6), δ: 9.86 (s, 1H), 8.29 (d, J = 5.5Hz, 1H), 7.95–7.73 (m, 1H) ,7.41(d,J=5.5Hz,1H),7.33–7.22(m,1H),7.22–7.10(m,2H),4.66(d,J=12.9Hz, 1H), 4.51 (d, J=13.4Hz, 1H), 3.70–3.46 (m, 1H), 3.40 (t, J=11.4Hz, 1H), 2.93–2. 75(m,1H),2.19–2.01(m,1H),2.00–1.89(m,1H),1.88–1.72(m,1H),1.58(m,1H); 13C NMR (150MHz, DMSO-d6), δ: 172.34, 163.16, 158.22, 156.51, 154.32 (d, J = 245.1Hz), 135.73, 126.28 (d, J = 11.6Hz), 126.02 (d, J = 7.6Hz), 125.01, 124.80, 124.30, 115.99 (d, J = 19.4Hz), 112.90, 48.48, 46.85, 42.78, 28.25, 24.70; HRMS (ESI), m / Z: theoretical value C 18 H 17 ClFN4OS[M+H] + 391.0796, measured value 391.0796.

[0065] Example 13: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-chlorophenyl)piperidine-3-carboxamide (Ib-4): white solid powder, yield 76.9%; 1 H NMR (600MHz, DMSO-d6), δ: 10.20 (s, 1H), 8.29 (d, J = 5.5Hz, 1H), 7.63 (d, J = 8.9 Hz,2H),7.41(d,J=5.5Hz,1H),7.39–7.28(m,2H),4.67(d,J=13.0Hz,1H),4.53 (d,J=13.3Hz,1H),3.61–3.44(m,1H),3.36(t,J=11.7Hz,1H),2.78–2.60(m,1H ),2.25–2.03(m,1H),2.00–1.88(m,1H),1.88–1.75(m,1H),1.70–1.49(m,1H); 13 CNMR (150MHz, DMSO-d6), δ: 172.03, 163.16, 158.21, 156.50, 138.36, 135.74, 129.11, 127.33, 124.31, 121.26, 112.90, 48.42, 46.85, 43.45, 28.17, 24.74; HRMS (ESI), m / Z: theoretical value C 18 H 17 Cl2N4OS[M+H] + 407.0500, measured value 407.0503.

[0066] Example 14: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-methylphenyl)piperidine-3-carboxamide (Ib-5): white solid powder, yield 71.8%; 1H NMR (600MHz, DMSO-d6), δ: 9.96 (s, 1H), 8.29 (d, J = 5.5Hz, 1H), 7.47 (d, J = 8.4Hz ,2H),7.41(d,J=5.5Hz,1H),7.11(d,J=8.3Hz,2H),4.68(d,J=13.2Hz,1H),4.55 (d,J=13.3Hz,1H),3.57–3.41(m,1H),3.37(m,1H),2.73–2.59(m,1H),2.25(s,3 H),2.15–1.99(m,1H),1.98–1.88(m,1H),1.87–1.73(m,1H),1.67–1.43(m,1H); 13 C NMR (150MHz, DMSO-d6), δ: 171.60, 163.16, 158.19, 156.52, 136.91, 135.75, 132.69, 129.56, 124.32, 119.75, 112.89, 48.58, 46.78, 43.47, 28.27, 24.80, 20.93; HRMS (ESI), m / Z: theoretical value C 19 H 20 ClN4OS[M+H] + 387.1046, measured value 387.1046.

[0067] Example 15: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-methoxyphenyl)piperidine-3-carboxamide (Ib-6): white solid powder, yield 81.4%; 1 H NMR (600MHz, DMSO-d6), δ: 9.92 (s, 1H), 8.29 (d, J = 5.5Hz, 1H), 7.59–7.47 (m, 2 H),7.41(d,J=5.5Hz,1H),6.97–6.80(m,2H),4.68(d,J=13.1Hz,1H),4.55(d, J=13.3Hz,1H),3.72(s,3H),3.54–3.41(m,1H),3.37(s,1H),2.70–2.58(m,1H) ),2.16–2.00(m,1H),1.98–1.88(m,1H),1.88–1.74(m,1H),1.69–1.47(m,1H); 13CNMR (150MHz, DMSO-d6), δ: 171.30, 163.15, 158.18, 156.52, 155.70, 135.74, 132.55, 124.31, 121.27, 114.29, 112.88, 55.62, 48.63, 46.85, 43.41, 28.27, 24.82; HRMS (ESI), m / Z: theoretical value C 19 H 20 ClN4O2S[M+H] + 403.0995, measured value 403.0995.

[0068] Example 16: 1-(2-chlorothiopheno[3,2-d]pyrimidin-4-yl)-N-(3,4-dimethoxyphenyl)piperidine-3-carboxamide (Ib-7): white solid powder, yield 78.4%; 1 H NMR (600MHz, DMSO-d6), δ: 9.92 (s, 1H), 8.29 (d, J = 5.5Hz, 1H), 7.41 (d, J = 5.5Hz, 1H), 7.32 (d ,J=2.4Hz,1H),7.09(dd,J=8.7,2.4Hz,1H),6.88(d,J=8.7Hz,1H),4.69(d,J=13.2Hz,1H),4. 56(d,J=13.3Hz,1H),3.72(s,3H),3.71(s,3H),3.53–3.40(m,1H),3.33(t,J=12.9Hz,1H),2. 71–2.57(m,1H),2.17–2.00(m,1H),1.99–1.88(m,1H),1.88–1.73(m,1H),1.68–1.47(m,1H); 13 C NMR (150MHz, DMSO-d6), δ: 171.36, 163.16, 158.18, 156.52, 148.95, 145.30, 135.74, 133.02, 124.32, 112.88, 112.45, 111.62, 104.85, 56.16, 55.78, 48.61, 46.87, 43.48, 28.29, 24.82; HRMS (ESI), m / Z: theoretical value C 20 H 22 ClN4O3S[M+H] + 433.1101, measured value 433.1102.

[0069] Example 17: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(1-adamantyl)piperidine-3-carboxamide (Ib-8): white solid powder, yield 83.8%; 1 H NMR (600MHz, DMSO-d6), δ: 8.29 (d, J = 5.5Hz, 1H), 7.40 (m, 2H), 4.49 (t, J = 13.7Hz, 2H), 3.34 –3.19(m,2H),2.45–2.30(m,1H),2.08–1.81(m,11H),1.77–1.55(m,7H),1.54–1.37(m,1H); 13 C NMR (150MHz, DMSO-d6), δ: 172.26, 163.10, 158.09, 156.52, 135.67, 124.30, 112.78, 51.08, 48.87, 46.81, 43.00, 41.40, 36.48, 29.25, 28.37, 24.79; HRMS (ESI), m / Z: theoretical value C 22 H 28 ClN4OS[M+H] + 431.1672, measured value 431.1673.

[0070] Example 18: In vitro antitumor cell activity of thieno[3,2-d]pyrimidine compounds containing piperidine amide structures.

[0071] Human lung adenocarcinoma cell line A549 and human liver cancer cell line HepG2 in logarithmic growth phase were selected, and the in vitro antitumor activity of the target compounds in Examples 1 to 17 was tested using the MTT assay.

[0072] (1) After cell resuscitation and stabilization through 2-3 passages, cells were digested from the bottom of the culture flask using trypsin solution (0.25%). The digestion solution was poured into a centrifuge tube, followed by the addition of culture medium to terminate the digestion. The centrifuge tube was centrifuged at 800 rpm for 10 min, the supernatant was discarded, and 5 mL of culture medium was added. The cells were mixed by pipetting, and 10 μL of the cell suspension was added to a cell counting chamber to count the cells, adjusting the cell concentration to 10⁴ cells / well. 100 μL of cell suspension was added to each well of the 96-well plate except for well A1, which was a blank well. The 96-well plate was then incubated in an incubator for 24 h.

[0073] (2) Dissolve the test sample in 50 μL of dimethyl sulfoxide, then add an appropriate amount of culture medium to dissolve the sample into a 2 mg / mL solution, and then dilute the sample in a 24-well plate to 20, 4, 0.8, 0.16, and 0.032 μg / mL.

[0074] Add cells to 3 wells at each concentration. The cells in the outer two rows and two columns are more susceptible to environmental influences and are suitable for use as blank cells. Incubate the 96-well plate in an incubator for 72 hours.

[0075] Discard the drug-containing culture medium in the 96-well plate, wash the cells twice with phosphate-buffered saline (PBS), add 100 μL of MTT (tetrazazole) (0.5 mg / mL) to each well, and incubate for 4 hours. Discard the MTT solution and add 100 μL of dimethyl sulfoxide. Shake on a magnetic oscillator to fully dissolve the surviving cells and the MTT reaction product, formazan. Measure the results using a microplate reader. The IC50 of the drug can be determined using the Bliss assay. 50 The results of the compounds' inhibitory activity against human lung adenocarcinoma A549 and human liver cancer cells HepG2 are shown in Table 1.

[0076] Table 1. Inhibitory activity of compounds against human lung adenocarcinoma A549 and human hepatocellular carcinoma HepG2.

[0077]

[0078] As can be clearly seen from Table 1, the compounds of general formula (I) protected by this invention exhibit good inhibitory activity against human lung adenocarcinoma cells A549 and human liver cancer cell line HepG2 in vitro. These compounds show promising potential for the development and application of antitumor drugs.

[0079] Although the invention has been described with reference to specific embodiments, modifications and equivalent variations will be apparent to those skilled in the art, and they are all included within the scope of the invention.

Claims

1. Thiophene[3,2-d]pyrimidine compounds containing a piperidine amide structure and their pharmaceutically acceptable salts, characterized in that, It has a general structural formula as shown in (I): in: R is selected from adamantyl alkyl group, C6-C 10 Yuanfangyi or C6-C 10 Heteroaryl, wherein the aryl group is replaced by any 1 to 3 identical or different R1 groups; R1 is selected from hydrogen, halogen, C1-C6 alkyl, or C1-C6 alkoxy.

2. The thieno[3,2-d]pyrimidine compound containing a piperidine amide structure and its pharmaceutically acceptable salt according to claim 1, characterized in that, Having a general structural formula as shown in (Ia) or (Ib): in: R is selected from adamantyl, phenyl, or pyridyl, wherein the phenyl or pyridyl group is substituted by any 1 to 3 identical or different R1 groups; R1 is selected from hydrogen, fluorine, chlorine, methyl, or methoxy.

3. The thiopheno[3,2-d]pyrimidine compound containing a piperidine amide structure and its pharmaceutically acceptable salt according to claim 2, characterized in that, It has the following compounds: 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-phenylpiperidin-4-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-fluorophenyl)piperidin-4-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(2-fluorophenyl)piperidin-4-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-chlorophenyl)piperidin-4-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-methylphenyl)piperidin-4-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-methoxyphenyl)piperidine-4-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(3,4-dimethoxyphenyl)piperidine-4-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(1-adamantyl)piperidin-4-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(2,4-difluorophenyl)piperidin-4-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-phenylpiperidin-3-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-fluorophenyl)piperidine-3-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(2-fluorophenyl)piperidine-3-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-chlorophenyl)piperidine-3-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-methylphenyl)piperidin-3-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(4-methoxyphenyl)piperidine-3-carboxamide 1-(2-chlorothiopheno[3,2-d]pyrimidin-4-yl)-N-(3,4-dimethoxyphenyl)piperidine-3-carboxamide 1-(2-chlorothieno[3,2-d]pyrimidin-4-yl)-N-(1-adamantyl)piperidine-3-carboxamide.

4. A pharmaceutical composition, characterized in that, The active ingredient and pharmaceutically acceptable excipient comprise a thieno[3,2-d]pyrimidine compound containing a piperidine amide structure as described in claim 1, 2 or 3, or a pharmaceutically acceptable salt thereof.

5. The use of the thiopheno[3,2-d]pyrimidine compound containing the piperidine amide structure as claimed in claim 1, 2 or 3, and its pharmaceutically acceptable salt, or the pharmaceutical composition of claim 4, in the preparation of a medicament for treating and / or preventing cancer.

6. The application according to claim 5, characterized in that, The cancers mentioned are lung cancer and liver cancer.

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