Pyridyl piperazine derivative as well as preparation method and application thereof

Abstract

The invention provides a pyridyl piperazine derivative as well as a preparation method and application thereof, and belongs to the technical field of medicinal chemistry. The structure of the pyridyl piperazine derivative is shown as a formula I in the specification. The pyridyl piperazine derivative with a novel structure is successfully synthesized through a large number of screening, the compound is found to be capable of effectively inhibiting proliferation of colon cancer cells, and the inhibiting effect is superior to that of a positive compound AZ191. The compound disclosed by the invention has a wide application prospect in preparation of medicines for preventing and / or treating tumors. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical chemistry, and specifically relates to a pyridylpiperazine derivative and a preparation method and application thereof. Background Art

[0002] Malignant tumors are one of the main causes of death in the world and have become a major disease that seriously endangers human life and health and restricts social and economic development. Lung cancer, breast cancer, stomach cancer, liver cancer, esophageal cancer, colorectal cancer, cervical cancer, prostate cancer, and lymphoma are the main common malignant tumors in my country.

[0003] Colon cancer is the third most common malignancy and the second most deadly malignancy. It is closely related to environmental and genetic risk factors. In recent years, the incidence and mortality rates have been increasing rapidly. The operation is difficult and the disease is prone to metastasis. Epidemiological studies have shown that an unbalanced diet (high in saturated fat and low in vegetables and fruits), lack of exercise leading to abdominal fat accumulation, excessive obesity, and a history of malignant tumors in first-degree family members are risk factors for colon cancer.

[0004] In recent years, although great progress has been made in chemotherapy for colon cancer, there are still some defects, such as: the commonly used anticancer drugs have a single mechanism of action, large toxic side effects, and long-term use can easily lead to chemotherapy resistance, which greatly reduces the treatment effect. According to statistics, about 90% of chemotherapy failures are related to chemotherapy resistance. Therefore, it is of great significance to develop new drugs for the treatment of colon cancer.

[0005]

[0006] AZ191 is a known compound with certain anti-tumor effects, but its ability to inhibit tumor cell proliferation cannot meet the needs and needs to be further improved. Summary of the invention

[0007] The object of the present invention is to provide a pyridylpiperazine derivative and a preparation method and use thereof.

[0008] The present invention provides a compound, a stereoisomer thereof, a deuterated substance thereof or a pharmaceutically acceptable salt thereof, wherein the structure of the compound is shown in Formula I:

[0009]

[0010] Where X1 is N or CR a , X2 is N or CR b , X3 is N or CR c , X4 is N or CR d , X5 is N or CR e , R a , R b, R c , R d , R e Each independently selected from hydrogen, C 1-8 Alkyl or C 1-8 Alkoxy;

[0011] R2 is selected from hydrogen, C 1-8 Alkyl or C 1-8 Alkoxy;

[0012] R1 is selected from hydrogen, C 1-8 Alkyl, COOR4, COR4, R4 is selected from C 1-8 alkyl.

[0013] Furthermore, the structure of the compound is shown in Formula II:

[0014]

[0015] Where X1 is N or CR a , X2 is N or CR b , X3 is N or CR c , X4 is N or CR d , X5 is N or CR e , R a , R b , R c , R d , R e Each independently selected from hydrogen, C 1-4 Alkyl or C 1-4 Alkoxy;

[0016] R2 is selected from hydrogen, C 1-4 Alkyl or C 1-4 Alkoxy;

[0017] R1 is selected from hydrogen, C 1-4 Alkyl, COOR4, COR4, R4 is selected from C 1-4 alkyl.

[0018] Furthermore, the structure of the compound is shown in Formula III:

[0019]

[0020] Wherein R1 is selected from hydrogen, C 1-4 Alkyl, COOR4, COR4, R4 is selected from C 1-4 alkyl.

[0021] Furthermore, the R4 is tert-butyl.

[0022] Furthermore, the compound is selected from one of the following compounds:

[0023]

[0024] The present invention also provides a pharmaceutical composition, which is a preparation prepared by using the above-mentioned compound, its stereoisomer, its deuterated product or its pharmaceutically acceptable salt as an active ingredient and adding pharmaceutically acceptable excipients.

[0025] Furthermore, the preparation is an oral preparation or an injection preparation.

[0026] The present invention also provides the use of the above-mentioned compound, its stereoisomer, its deuterated product or its pharmaceutically acceptable salt in preparing a drug for preventing and / or treating tumors.

[0027] Furthermore, the tumor is colon cancer, lung cancer, breast cancer, stomach cancer, liver cancer, esophageal cancer, cervical cancer, prostate cancer or lymphoma.

[0028] Furthermore, the tumor is colon cancer.

[0029] Definitions of terms used in the present invention: Unless otherwise stated, the initial definitions provided for groups or terms in this document apply to the groups or terms throughout the specification; for terms that are not specifically defined in this document, the meaning that a person skilled in the art can give them should be given based on the disclosure and context.

[0030] The minimum and maximum carbon atom content in a hydrocarbon group is indicated by a prefix, for example, the prefix C a～b Alkyl refers to any alkyl group containing from "a" to "b" carbon atoms. 1～4 Alkyl refers to a straight or branched chain alkyl group containing 1, 2, 3 or 4 carbon atoms. 1～6 Alkoxy refers to straight or branched chain alkoxy groups containing 1, 2, 3 or 4 carbon atoms, and so on.

[0031] The present invention successfully synthesized a class of novel pyridylpiperazine derivatives through extensive screening, and found that the compound can effectively inhibit the proliferation of colon cancer cells, and the inhibitory effect is better than that of the positive compound AZ191. The compound of the present invention has broad application prospects in the preparation of drugs for preventing and / or treating tumors.

[0032] The preparation method of the compound of the invention is simple and easy to implement and is suitable for industrial production.

[0033] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0034] The above contents of the present invention are further described in detail below through specific implementation methods in the form of examples. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. DETAILED DESCRIPTION

[0035] The raw materials and equipment used in the present invention are all known products, which are obtained by purchasing commercially available products.

[0036] Example 1. Synthesis of Compounds 14 and 17

[0037] Intermediate Series 4 Compound II-1

[0038]

[0039] Weigh 0.193g (1mmol) of series 4 compound I and 0.102g (0.5mmol) of phenylboronic acid ester. Place in a round-bottom flask, add 100mg of Pd(PPh3)4, prepare 1.5ml of K2CO3 solution (200mg) and add it to the reaction system, add 15ml of 1,4-dioxane as solvent, use Schlenk tube to extract air in the bottle and fill it with nitrogen protection, repeat 6 times, and you can get a reaction system under nitrogen protection. Stir the reaction at 100℃ for 6h, and the reaction liquid is black. Monitor the reaction liquid by TLC (developing solvent is ethyl acetate (EA): petroleum ether (PE) = 1:10), the raw material disappears, the reaction is complete, and the reaction is stopped. Water was added to terminate the reaction, and the reaction solution was extracted with ethyl acetate (3×40 ml). The organic phase was washed with brine and dried over anhydrous MgSO4. The filtrate was rotary evaporated to obtain a crude product, which was purified by column chromatography (ethyl acetate (EA): petroleum ether (PE) = 1:15) to obtain 73 mg of a white solid with a yield of 76.8%. 1 HNMR(400MHz,DMSO-d6)δ9.14(s,2H),7.90-7.77(m,2H),7.63-7.45(m,3H). 13 C NMR (400MHz, DMSO-d6) δ 162.28, 160.64, 152.56, 142.63, 133.52, 130.71, 124.02. Intermediate series 4 compound II-2

[0040] According to the synthesis method of intermediate series 4 compound II-1, pyridyl borate was used as raw material to prepare series 4 compound II-2, a yellow solid, with a yield of 79.6%. 1H NMR(400MHz,DMSO-d6)δ9.26(s,2H),8.77–8.71(m,2H),7.95–7.83(m,2H).13C NMR(101MHz,DMSO-d6)δ160.86,159.04,150.98,140.52,130.25,121.86.

[0041]

[0042] Compound 17

[0043]

[0044] Weigh compound A (1mmol) and series 4 compound II-1 (1mmol), place in a round-bottom flask, add 3mmol p-toluenesulfonic acid hydrate, 5ml water, and 15ml Diox. Stir and react at 100℃ for 12h, the reaction solution is yellow. Monitor the reaction solution by TLC (developing solvent is ethyl acetate (EA): petroleum ether (PE) = 1:5), the raw material disappears, the reaction is complete, and the reaction is stopped. Add sodium bicarbonate to adjust the alkalinity, extract the reaction solution with ethyl acetate (3×50ml), wash the organic phase with brine, dry with anhydrous MgSO4, and spin dry to obtain series 4 compound III-1 (i.e. compound 17) as a yellow solid. The yield is 15.2%. 1 H NMR(400MHz,DMSO-d6)δ9.57(s,1H),8.77(s,2H),7.72–7.66(m,2H),7.65–7.59(m,2H),7.49–7.43(m ,2H),7.39–7.31(m,1H),6.96–6.90(m,2H),3.46(t,J=4.7Hz,4H),3.02(t,J=5.2Hz,4H),1.42(s,9H). 13 C NMR (400MHz, DMSO-d6) δ159.89,156.17,154.30,146.61,135.32,129.56,125.96,123.89,120.76,117.14,79.41,49.79(2C),28.54.

[0045]

[0046] Compound 14

[0047]

[0048] Weigh 0.23mmol of series 4 compound III-1, add 10ml Diox·HCl, react at room temperature for 3h, the reaction solution turns red. Monitor the reaction solution by TLC (developing solvent is ethyl acetate (EA): petroleum ether (PE) = 1:1), the raw material disappears, the reaction is complete, and the reaction is stopped. Vacuum spin off the dioxane hydrochloric acid solution, add 1M NaOH to adjust the alkalinity, extract the reaction solution with ethyl acetate (3×50ml), wash the organic phase with brine, dry with anhydrous MgSO4, and recrystallize with petroleum ether to obtain series 4 compound VI-1 (i.e. compound 14) as a yellow solid. The yield is 20.8%. 1 H NMR (400MHz, DMSO-d6) δ9.52(s,1H),8.76(s,2H),7.70–7.67(m,2H),7.62–7.59(m,2H),7.47(d,J=7. 6Hz, 2H), 7.35 (t, J=7.4Hz, 1H), 3.00 (dd, J=6.4, 3.5Hz, 4H), 2.87 (dd, J=6.2, 3.8Hz, 4H), 1.24 (s, 1H). 13 C NMR(400MHz,DMSO-d6)δ159.93,156.15,147.35,135.36,132.87,129.55,12 7.70,125.94,123.81,120.87,116.39,109.69,50.37,45.91.HRMS(ESI)m / z for[M+H] + :Exact Mass:331.1797; Found:332.1883 Example 2, Synthesis of Compound 32

[0049]

[0050] Weigh series 9 compound I (0.05 mol) and Pd(Amphos)Cl2 (0.075 mol) and place them in a three-necked flask. Add 100 ml of DMF, then add 4-(4-methylpiperazine)aniline (0.064 mol) and Cs2CO3 (0.1 mol). Use a Schlenk tube to extract the air in the flask and fill it with nitrogen protection. After repeating 6 times, a reaction system under nitrogen protection can be obtained. Reflux the reaction at 145°C. The reaction solution is monitored by TLC (developing solvent is ethyl acetate (EA): petroleum ether (PE) = 1:1). The raw material disappears and the reaction is complete, and the reaction is stopped. Add water to terminate the reaction, extract the reaction solution with ethyl acetate (3×50 ml), wash the organic phase with brine, dry it with anhydrous MgSO4, and the crude product obtained after rotary evaporation of the filtrate is subjected to column chromatography (ethyl acetate (EA): petroleum ether (PE) = 1:3) to obtain compound 32 as a light yellow solid. The yield is 26%.1 H NMR(400MHz, DMSO-d6)δ9.52(s,1H),8.76(d,J=1.0Hz,2H),7.71–7.67(m,2H),7.61(dd,J=9.2,2.8Hz,2H),7.4 6(dd,J=8.4,7.0Hz,2H),7.38–7.33(m,1H),6.92–6.89(m,2H),3.08(t,J=5.0Hz,4H),2.53(s,4H),2.26(s,3H). 13 CNMR(101MHz,DMSO-d6)δ159.92,156.15,156.06,146.61,135.35,129.55,127.71,125.95,123.83,120.85,116.40,55.38,54.95,49.10.

[0051] The following is the preparation method of the control compound.

[0052] Comparative Example 1: Synthesis of Compounds 1-4

[0053] Intermediate Series 1 Compound II

[0054]

[0055] Weigh 1.3g Zn powder, 220mg CoBr2, 6ml acetonitrile, put into a 50ml three-necked bottle, then add 250μl allyl chloride and 100μl trifluoroacetic acid. The solution changes from green to blue and is stirred at room temperature for three minutes. 1.22ml (10mmol) 4-fluorobromobenzene and 1.15g (7.5mmol) of series 1 compound I are added in sequence, the solution changes from blue to red-black, and stirred at 50℃ for 4h. The reaction solution is monitored by TLC (developing solvent is ethyl acetate (EA): petroleum ether (PE) = 5:1), the raw material disappears, the reaction is complete, and the reaction is stopped. 20ml saturated ammonium chloride solution is added to the reaction solution, extracted with dichloromethane (3×70ml), washed with brine, dried with MgSO4, and the crude product obtained after rotary evaporation of the filtrate is subjected to column chromatography (eluent is EA:PE = 1:9) to obtain 1.1g series 1 compound II, a white solid, with a yield of 83.86%. 1 H NMR (400MHz, Chloroform-d) δ8.78 (d, J = 4.8Hz, 2H), 8.49–8.42 (m, 2H), 7.20–7.13 (m, 3H). 13C NMR (400MHz, Chloroform-d) δ163.76,151.19,129.22,120.36,116.53,49.33,46.83.

[0056] Compound 1

[0057]

[0058] Weigh 50 mg (0.287 mmol) of series 1 compound II and 26.88 mg (0.315 mmol) of phenylpiperazine and place them in a 20 ml thick-walled pressure bottle. Add 282.4 mg (0.861 mmol) of Cs2CO3 and 9 ml of DMSO. Seal the tube and react at 100°C for 24 hours. The solution is black. Monitor the reaction solution by TLC (developing solvent is ethyl acetate (EA): petroleum ether (PE) = 10:1). The raw material disappears and the reaction is complete. Stop the reaction. After the bottle is cooled, add appropriate amount of water to the reaction system, extract the reaction solution with ethyl acetate (3×50 ml), wash the organic phase with brine, dry it with anhydrous MgSO4, and the crude product obtained after rotary evaporation of the filtrate is subjected to column chromatography (eluent is EA:PE = 1:15) to obtain 7 mg of compound III-1 as a yellow solid with a yield of 10.19%. 1 H NMR(400MHz,Chloroform-d)δ8.67(d,J=4.8Hz,2H),8.33–8.29(m,2H),7.25(dd,J=8.6,7.2Hz ,2H),7.03–6.94(m,5H),6.86(t,J=7.5Hz,1H),3.49–3.40(m,4H),3.31(dd,J=6.4,3.8Hz,4H). 13 C NMR(400MHz,Chloroform-d)δ163.50,156.09,151.57,128.46,128.34,117.04,114.16,47.10.HRMS(ESI)m / z for[M+H] + :ExactMass:316.1688;Found:317.1764

[0059] Compound 2

[0060]

[0061] According to the synthesis method of compound 1, pyridylpiperazine was used as raw material to prepare compound 2. White solid, the yield was 26.9%. 1H NMR(400MHz,DMSO-d6)δ8.80(d,J=4.8Hz,2H),8.31–8.26(m,2H),8.20(d,J=5.7Hz,2H),7.30(t,J=4.8Hz ,1H),7.12–7.08(m,2H),6.90(d,J=6.0Hz,2H),3.52(dd,J=7.1,3.7Hz,4H),3.46(dd,J=6.9,3.7Hz,4H). 13 C NMR(400MHz,Chloroform-d)δ157.15,154.95,152.24,148.95,129.49,128.76,118.15,114.87,108.21,47.53,45.70.HRMS(ESI)m / zfor[M+H] + :Exact Mass:317.1640;Found:318.1711

[0062] Compound 3

[0063]

[0064] According to the synthesis method of compound 1, cyclohexylpiperazine was used as raw material to prepare compound 3. Yellow solid, the yield was 10.5%. 1 H NMR(600MHz,Chloroform-d)δ8.73(d,J=4.8Hz,2H),8.35–8.31(m,2H),7.07(t, J=4.8Hz,1H),7.00–6.96(m,2H),3.41(s,4H),2.83(s,4H),1.38–1.15(m,10H). 13 CNMR(300MHz,Chloroform-d)δ164.69,157.10,152.81,129.37,128.23,117.96,114.87,63.89,48.72,48.11,28.64,26.12,25.75.DEPT135 NMR(400MHz,Chloroform-d)δ157.10,129.37,117.97,114.88,63.94,48.70,48.05,28.59,26.09,25.73.HRMS(ESI)m / zfor[M+H] + :Exact Mass:322.2157;Found:323.2236

[0065] Compound 4

[0066]

[0067] According to the synthesis method of compound 1, tert-butylpiperazine was used as a raw material to prepare compound 4. The compound 4 was a light yellow solid with a yield of 20.6%. 1 H NMR(600MHz,DMSO-d6)δ8.79(d,J=4.8Hz,2H),8.25–8.23(m,2H),7.28(t,J=4.8H z,1H),7.04–6.99(m,2H),3.26–3.24(m,4H),2.64(t,J=5.0Hz,4H),1.05(s,9H). 13 C NMR(151MHz,DMSO-d6)δ163.98,157.88,153.12,129.29,127.12,118.92,114.41,53.67,48.36,45.76,26.11.HRMS(ESI)m / z for[M+H] + :Exact Mass:296.2001;Found:297.2080

[0068] Comparative Example 2: Synthesis of Compounds 8-13

[0069] Intermediate Series 3 Compound II-1

[0070]

[0071] Weigh 0.9g (6.38mmol) of series 3 compound I and 1.067g (6.59mmol) of phenylpiperazine with different substituents, put them into a round-bottom flask, add 4.55g K2CO3 (18mmol) and 30ml DMSO, and react at 120℃ for 12h. The reaction liquid is yellow. The reaction liquid is monitored by TLC (developing solvent is ethyl acetate (EA): petroleum ether (PE) = 1:15). The raw material disappears and the reaction is complete, and the reaction is stopped. After the bottle is cooled, add a large amount of water, extract the reaction liquid with ethyl acetate (3×90ml), wash the organic phase with brine, dry it with anhydrous MgSO4, and the crude product obtained after rotary evaporation of the filtrate is subjected to column chromatography (eluent DCM) to obtain 1.65g of series 3 compound II-1, a yellow solid, with a yield of 88.7%. 1 HNMR(400MHz,DMSO-d6)δ8.12–8.04(m,2H),7.27–7.22(m,2H),7.11–7.06(m,2H),7 .02–6.96(m,2H),6.82(tt,J=7.2,1.0Hz,1H),3.66–3.58(m,4H),3.32–3.28(m,4H). 13C NMR (400MHz, DMSO-d6) δ155.06,151.03,137.44,129.49,126.22,119.65,115.99,113.14,48.22,46.60.

[0072] Intermediate Series 3 Compound II-2

[0073] According to the synthesis method of intermediate series 3 compound II-1, pyridylpiperazine was used as the raw material to prepare series 3 compound II-2, a yellow solid. The yield was 79.2%. 1H NMR (400MHz, DMSO-d6) δ8.21–8.17 (m, 2H), 8.12–8.06 (m, 2H), 7.07–7.01 (m, 2H), 6.87–6.80 (m, 2H), 3.68–3.61 (m, 4H), 3.57–3.48 (m, 4H). 13 CNMR(400MHz,DMSO-d6)δ154.75,154.36,150.30,137.31,126.25,112.80,108.48,45.83,44.90.

[0074] Intermediate Series 3 Compound II-3

[0075] According to the synthesis method of intermediate series 3 compound II-1, cyclohexylpiperazine was used as raw material to prepare series 3 compound II-3, a yellow solid, with a yield of 82.4%. 1 HNMR(400MHz, DMSO-d6)δ8.08–7.98(dd,2H),7.04–6.97(dd,2H),3.42(m,4H),2.63–2.55(m,4H),1.80–1.03(m,10H). 13 C NMR 400MHz, DMSO-d6)δ155.24,137.15,126.20,112.93,62.94,48.61,47.25,28.76,26.34,25.73.

[0076] Intermediate Series 3 Compound II-4

[0077] According to the synthesis method of intermediate series 3 compound II-1, series 3 compound II-4 was prepared using tert-butylpiperazine as raw material, as a yellow solid, with a yield of 78.8%. 1 H NMR (400MHz, Chloroform-d) δ8.15–8.09(dd,2H),6.85–6.77(dd,2H),3.47(s,4H),2.76(s,4H),1.16(s,9H).13 C NMR (400MHz, Chloroform-d) δ166.97,147.56,125.98,112.52,72.05,47.46,45.48,25.69.

[0078] Intermediate Series 3 Compound II-5

[0079] According to the synthesis method of intermediate series 3 compound II-1, series 3 compound II-5 was prepared using BOC-piperazine as raw material, as a yellow solid, with a yield of 82.8%. 1 H NMR (400MHz, Chloroform-d) δ8.19–8.05(dd,2H),6.89–6.75(dd,2H),3.70–3.57(m,4H),3.42(m,4H),1.49(s,9H). 13 C NMR (400MHz, DMSO-d6) δ158.39,133.64,120.70,117.14,112.14,79.40,49.83,28.54.

[0080] Intermediate Series 3 Compound III-1

[0081]

[0082] Weigh 1.5g (5mmol) of series 3 compound II-1, 0.85g (15.9mmol) of ammonium chloride into a round-bottom flask, add 20ml of water and ethyl acetate respectively, stir the reaction system after dissolution, and slowly add 2.8g (53mmol) of Fe powder while stirring. Let the reaction take place overnight at room temperature. Monitor the reaction solution by TLC (developing solvent is ethyl acetate (EA): petroleum ether (PE) = 1:3). When the raw material disappears, the reaction is complete and the reaction is stopped. Add sodium bicarbonate to adjust the alkalinity, extract the reaction solution with ethyl acetate (3×90ml), wash the organic phase with brine, dry it with anhydrous MgSO4, and spin dry to obtain 1.16g of black solid with a yield of 86.5%. 1 H NMR(400MHz,DMSO-d6)δ7.26–7.19(m,2H),7.00–6.96(m,2H),6.82–6.77(m,1H),6.7 6–6.73(m,2H),6.54–6.49(m,2H),4.61(s,2H),3.25–3.22(m,4H),3.06–3.03(m,4H). 13C NMR (400MHz, DMSO-d6) δ151.51,142.93,142.71,129.42,119.42,118.72,116.00,115.21,51.06,49.01.

[0083] Intermediate series 3 compound III-2

[0084] According to the synthesis method of intermediate series 3 compound III-1, series 3 compound II-2 was used as raw material to prepare series 3 compound III-2, a black solid, with a yield of 81.3%. 1 HNMR(400MHz,DMSO-d6)δ8.21–8.15(m,2H),6.89–6.83(m,2H),6.77–6.71(m ,2H),6.56–6.46(m,2H),4.64(s,2H),3.46–3.39(m,4H),3.07–2.93(m,4H). 13 C NMR (400MHz, DMSO-d6) δ155.00,150.34,143.17,142.48,118.94,115.16,108.92,50.75,46.11.

[0085] Intermediate series 3 compound III-3

[0086] According to the synthesis method of intermediate series 3 compound III-1, series 3 compound II-3 was used as raw material to prepare series 3 compound III-3, a black solid. The yield was 71.8%. 1HNMR (400MHz, DMSO-d6) δ 6.68–6.64 (m, 2H), 6.49–6.44 (m, 2H), 4.60 (s, 2H), 2.88 (t, J = 4.8 Hz, 4H), 2.61 (t, J = 4.9 Hz, 4H), 1.84–1.13 (m, 11H). 13 C NMR (400MHz, DMSO-d6) δ142.96,142.51,118.26,115.19,63.07,51.24,49.11,28.79,26.37,25.74.

[0087] Intermediate Series 3 Compound III-4

[0088] According to the synthesis method of intermediate series 3 compound III-1, series 3 compound II-4 was used as raw material to prepare series 3 compound III-4, a black solid, with a yield of 61.4%. 1HNMR (400MHz, Chloroform-d) δ6.88–6.78(m,2H),6.69–6.55(m,2H),4.62(s,2H),3.10(t,J=4.9Hz,4H),2.78(t,J=5.0Hz,4H),1.14(s,9H). 13 C NMR (400MHz, Chloroform-d) δ169.97, 147.56, 125.98, 112.52, 72.05, 47.46, 45.48, 25.69. Intermediate series 3 compound III-5

[0089] According to the synthesis method of intermediate series 3 compound III-1, series 3 compound II-5 was used as raw material to prepare series 3 compound III-5, a black solid, with a yield of 74.2%. 1 HNMR(400MHz,DMSO-d6)δ8.40(d,J=4.7Hz,2H),7.62–7.53(m,2H),6.94–6.88(m,2H),4.63(s,2H),3.46(t,J=5.2Hz,4H),3. 00(t,J=5.2Hz,4H),1.42(s,9H).13CNMR(400MHz,DMSO-d6)δ163.34,133.64,120.70,117.14,112.14,79.40,49.83,28.54.

[0090] Compound 8

[0091]

[0092] Weigh 250 mg (1 mmol) of intermediate series 3 compound III-1 and 114 mg (1 mmol) of 2-chloropyrimidine, place in a round-bottom flask, add 570 mg (3 mmol) of p-toluenesulfonic acid hydrate, 5 ml of water, and 15 ml of Diox. Stir and react at 100°C for 12 hours, and the reaction solution is yellow. Monitor the reaction solution by TLC (developing solvent is ethyl acetate (EA): petroleum ether (PE) = 1:5), the raw material disappears, the reaction is complete, and the reaction is stopped. Add sodium bicarbonate to adjust the alkalinity, extract the reaction solution with ethyl acetate (3×50 ml), wash the organic phase with brine, dry it with anhydrous MgSO4, and spin dry to obtain 61.5 mg of brown solid, with a yield of 20.1%. 1H NMR(500MHz,Chloroform-d)δ8.38(d,J=4.9Hz,2H),7.50(d,J=8.9Hz,2H),7.30(t,J=7.9Hz,2H),7.19(s,1 H),7.02(m,4H),6.91(t,1H),6.67(t,J=4.8Hz,1H),3.38(m,J=7.2,3.7Hz,4H),3.33(m,J=7.1,3.5Hz,4H). 13 CNMR(400MHz Chloroform-d)δ160.43,158.05,151.09,147.08,130.93,129.22,121.67,120.26,117.54,116.48,112.07,50.34,49.44.HRMS(ESI)m / z for[M+H] + :Exact Mass:331.1797;Found:332.3000

[0093] Compound 9

[0094] According to the synthesis method of compound 8, series 3 compound III-2 was used as the raw material to obtain a yellow solid with a yield of 18.7%. 1 HNMR(400MHz,DMSO-d6)δ9.39(s,1H),8.41(d,J=4.8Hz,2H),8.19(d,J=6.5Hz,2H),7.60(d,J=9.1Hz,2H),6 .97–6.94(m,2H),6.91(dd,J=5.2,1.5Hz,2H),6.75(t,J=4.8Hz,1H),3.51–3.47(m,4H),3.19–3.15(m,4H). 13 C NMR(400MHz,DMSO-d6)δ160.66,158.40,155.07,149.75,146.32,133.56,120.79,116.87,112.15,108.90,49.33,45.93.HRMS(ESI)m / zfor[M+H] + :Exact Mass:332.1749;Found:333.1820

[0095] Compound 10

[0096] According to the synthesis method of compound 8, series 3 compound III-3 was used as the raw material, and the yellow solid was obtained with a yield of 16.2%. 1HNMR(400MHz,DMSO-d6)δ9.33(s,1H),8.39(d,J=4.8Hz,2H),7.56–7.49(m,2H),6.90–6.81(m,2H),6.73(t,J=4.8Hz,1H),3.02 (d,J=5.7Hz,4H),2.63(s,4H),1.77(d,J=26.5Hz,4H),1.58(d,J=12.3Hz,1H),1.21(dd,J=10.8,8.1Hz,4H),1.14–1.06(m,1H). 13 C NMR (400MHz, DMSO-d6) δ160.68, 158.38, 145.94 (d, J = 171.1Hz), 132.93, 120. 79,116.26,112.02,63.03,50.04,48.96,28.82,26.38,25.75.HRMS(ESI)m / z for[M+H] + :Exact Mass:337.2266;Found:338.2347

[0097] Compound 11

[0098] According to the synthesis method of compound 8, series 3 compound III-4 was used as the raw material to obtain a yellow solid with a yield of 15.3%.

[0099] 1 H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.40(d,J=4.8Hz,2H),7.57–7.53(m,2H),6.89– 6.85(m,2H),6.74(t,J=4.8Hz,1H),3.05(t,J=4.9Hz,4H),2.67(s,4H),1.06(s,9H). 13 C NMR(400MHz,DMSO-d6)δ160.68,158.39,146.70,132.93,120.83,116.18,112.02,50.14,45.86,26.04.HRMS(ESI)m / zfor[M+H] + :Exact Mass:311.2110;Found:312.2184

[0100] Compound 12

[0101]

[0102] Weigh 250 mg (1 mmol) of series 3 compound III-5 and 114 mg (1 mmol) of 2-chloropyrimidine, place in a round-bottom flask, add 570 mg (3 mmol) of p-toluenesulfonic acid hydrate, 5 ml of water, and 15 ml of Diox. Stir and react at 100°C for 12 hours, and the reaction solution is yellow. Monitor the reaction solution by TLC (developing solvent is ethyl acetate (EA): petroleum ether (PE) = 1:5), the raw material disappears, the reaction is complete, and the reaction is stopped. Add sodium bicarbonate to adjust the alkalinity, extract the reaction solution with ethyl acetate (3×50 ml), wash the organic phase with brine, dry it with anhydrous MgSO4, and spin dry to obtain 61.5 mg of brown solid, with a yield of 20.1%. 1 H NMR (400MHz, DMSO-d6) δ9.36 (s, 1H), 8.40 (d, J = 4.7Hz, 2H), 7.60–7.55 (m, 2H), 6.93–6.86 (m,2H),6.75(t,J=4.8Hz,1H),3.46(t,J=5.2Hz,4H),3.00(t,J=5.2Hz,4H),1.42(s,9H). 13 C NMR(400MHz,DMSO-d6)δ160.65,158.39,154.30,146.50,133.64,120.70,117.14,112.14,100.23,79.40,49.83,28.54.HRMS(ESI)m / z for[M+H] + :Exact Mass:355.2008;Found:356.2086

[0103] Compound 13

[0104]

[0105] According to the synthesis method of compound 8, series 3 compound III-5 was used as the raw material, and the yellow solid was obtained with a yield of 19.6%. 1 HNMR(400MHz,DMSO-d6)δ8.40(dd,J=6.8,4.7Hz,4H),7.59(dd,J=9.1,2.6,1.9Hz,2H),6 .99–6.92(m,2H),6.75(t,J=4.7Hz,1H),6.66(t,J=4.7Hz,1H),3.88(m,4H),3.12(m,4H). 13C NMR (400MHz, DMSO-d6) δ161.68,160.64,158.46,158.40,146.63,133.52,120.71,117.02,112.64,112.12,110.76,49.72,43.75.HRMS (ESI) m / z for[M+H] + :Exact Mass:333.1702;Found:334.1772

[0106] Comparative Example 3: Synthesis of Compounds 25 and 27

[0107] Intermediate Series 5 Compound II

[0108]

[0109] Weigh 10g (0.05mol) of 2,4-dichloro-5-aminopyrimidine and 530mg (0.075mol) of Pd(Amphos)Cl2 and place them in a three-necked flask. Add 100ml of DMF, then add 22ml (0.2mol) of ethyl acrylate and 13.9ml (0.1mol) of Et3N. Use a Schlenk tube to extract the air in the flask and fill it with nitrogen protection. After repeating 6 times, a reaction system under nitrogen protection can be obtained. Reflux the reaction at 145°C. Monitor the reaction solution by TLC (developing solvent is ethyl acetate (EA): petroleum ether (PE) = 1:1). If the raw material disappears, the reaction is complete and the reaction is stopped. Water was added to terminate the reaction, and the reaction solution was extracted with ethyl acetate (3×50 ml). The organic phase was washed with brine and dried over anhydrous MgSO4. The filtrate was rotary evaporated to obtain a crude product, which was purified by column chromatography (ethyl acetate (EA): petroleum ether (PE) = 1:3) to obtain 5.69 g of a yellow solid with a yield of 50%. 1 H NMR (400MHz, DMSO-d6) δ8.22(s,1H),7.87(d,J=15.2Hz,1H),6.84(d,J=15.2Hz,1H),6.39(s,2H),4.21(q,2H),1.23(t,3H). 13 CNMR (400MHz, DMSO-d6) δ 166.03, 149.98, 142.17, 141.04, 136.24, 123.53, 60.89, 14.61. Intermediate series 5 compound III

[0110]

[0111] Weigh 2 g of series 5 compound II and tri-n-butylphosphine, add acetic acid, and seal the tube for reaction at 110°C for 1 h. After the reaction solution is cooled, vacuum dry to remove acetic acid. After the solid is washed with saturated sodium bicarbonate, the reaction solution is extracted with ethyl acetate (3×50 ml), the organic phase is washed with brine, and dried over anhydrous MgSO4. The crude product obtained after rotary evaporation of the filtrate is subjected to column chromatography (ethyl acetate (EA): petroleum ether (PE) = 1:2-1:0) to obtain 0.4 g of yellow solid, with a yield of 25%. 1 H NMR (400MHz, DMSO-d6) δ12.29(s,1H),8.74(s,1H),7.91(d,J=9.9Hz,1H),7.07(d,J=9.9Hz,1H). 13 C NMR (101 MHz, DMSO-d6) δ 160.95, 152.48, 148.73, 144.01, 138.92, 133.42, 132.10. Intermediate series 5 compound IV

[0112]

[0113] Weigh 50 mg (0.225 mmol) of series 5 compound III, add 10 ml of anhydrous DMF, and add 26.6 mg (0.675 mmol) of NaH in batches after ice bath at -20°C for 5 min. After reacting at -20°C for 30 min, add 67.5 ul (0.6 mmol) of iodomethane, move the reaction bottle to room temperature, and continue to react for 2 h. After the reaction is completed, pour the reaction solution into ice water, extract the reaction solution with ethyl acetate (3×50 ml), wash the organic phase with brine, dry it with anhydrous MgSO4, and the crude product obtained after rotary evaporation of the filtrate is subjected to column chromatography (ethyl acetate (EA): petroleum ether (PE) = 1:2) to obtain 26 mg of white solid, with a yield of 60%. 1 HNMR (500MHz, CDCl3) δ8.72(s,1H),7.73(d,J=9.8Hz,1H),7.10(d,J=9.8Hz,1H),3.68(s,3H).

[0114] Compound 25

[0115]

[0116] Weigh 195 mg (1 mmol) of series 5 compound IV and 554 mg (2 mmol) of 1-Boc-4-(4-aminophenyl)piperazine into a three-necked flask. Add 984 mg (3 mmol) of CS2CO3, then weigh 45.75 mg (0.05 mmol) of Pd2(dba)3 and 62.2 mg (0.1 mmol) of BINAP, dissolve them in toluene, add them to the reaction system, and add 15 ml of toluene. Use a Schlenk tube to extract the air in the bottle and fill it with nitrogen protection. Repeat 6 times to obtain a reaction system under nitrogen protection. Reflux the reaction at 100°C. Monitor the reaction solution by TLC (developing solvent is ethyl acetate (EA): petroleum ether (PE) = 3:1). If the raw material disappears, the reaction is complete and the reaction is stopped. Water was added to terminate the reaction, and the reaction solution was extracted with ethyl acetate (3×50 ml). The organic phase was washed with brine and dried over anhydrous MgSO4. The filtrate was rotary evaporated to obtain a crude product, which was then purified by column chromatography (ethyl acetate (EA): petroleum ether (PE) = 1:1) to obtain a crude product, and then purified by column chromatography with dichloromethane (DCM): methanol (MeOH) = 60:1 to obtain 196 mg of a red solid with a yield of 45%. 1 HNMR (400MHz, DMSO-d6) δ9.52(s,1H),8.91(s,1H),7.71(d,J=9.8Hz,1H),7.67(d,J=8.5Hz,2H),7.00(d,J=9.7H z,1H),6.95(d,J=8.5Hz,2H),3.62(s,3H),3.48(d,J=5.1Hz,5H),3.07–2.99(m,4H),1.42(s,9H).HRMS(ESI)m / z for[M+H] + :Exact Mass:436.2223;Found:437.2299

[0117] Compound 27

[0118]

[0119] According to the synthesis method of compound 5 in Example 1, compound 27 was prepared as a red solid with a yield of 52%. 1 HNMR(500MHz,DMSO-d6)δ9.54(s,1H),9.46–8.98(m,1H),8.91(s,1H),7.70(dd,J=9.3,5.0Hz,3H), 7.00(d,J=9.7Hz,1H),6.96(d,J=8.7Hz,2H),3.63(s,3H),3.29(d,J=5.2Hz,4H),3.26–3.19(m,4H). 13C NMR (400MHz, DMSO-d6) δ159.51,156.23,147.35,145.33,143.11,138.64,134.43,131.11,127.50,120.05,117.26,46.87,43.18,29.00.

[0120] Comparative Example 4: Synthesis of Compound 22

[0121]

[0122] According to the synthesis method of series 3 compound 12, compound 22 was prepared according to the above route. It was a black solid with a yield of 66.8%. 1 H NMR (400MHz, DMSO-d6) δ8.73(s,2H),7.65(d,J=7.6Hz,2H),7.45(t,J=7.5Hz,2H),7.34(t,J=7.4Hz,1H) ,6.76(d,J=8.2Hz,2H),6.52(d,J=8.2Hz,2H),4.78(s,2H),3.91(t,J=5.1Hz,4H),2.98(t,J=5.1Hz,3H). 13 C NMR(400MHz,DMSO-d6)δ161.04,156.26,142.95,142.86,135.42,129.53,127.60,125.95,122.53,119.08,115.27,51.11,44.21.Exact Mass:331.1797; Found:332.1877

[0123] Comparative Example 5: Synthesis of Compounds 39 and 30

[0124] Intermediate Series 8 Compound II

[0125]

[0126] According to the above route, series 8 compound II was prepared as a white solid with a yield of 74.5%. 1 H NMR (400MHz, DMSO-d6) δ9.16(s,2H),7.11(s,2H),3.88(s,6H),3.71(s,3H). 13 CNMR(400MHz,DMSO-d6)δ159.13,158.49,154.06,138.88,132.78,128.36,105.11,60.58,56.68.

[0127] Compound 29

[0128]

[0129] According to the synthesis method of compound 6 in Example 1, series 8 compound II and 4-[4-(N-BOC)piperazine-1-yl]phenylboronic acid pinacol ester were used as raw materials. Yellow solid, yield 49.4%. 1 HNMR(400MHz,DMSO-d6)δ9.16(s,2H),8.35–8.23(m,2H),7.10(s,2H),7.09–7.05(m,2H) ,3.89(s,6H),3.71(s,3H),3.48(dd,J=6.5,3.8Hz,4H),3.31–3.27(m,4H),1.43(s,9H). 13 C NMR (400MHz, DMSO-d6) δ162.54,155.53,154.36,154.03,152.90,138.38,130.2 5,130.17,129.27,127.42,115.13,104.55,79.53,60.57,56.62,47.59,28.54.

[0130] Compound 30

[0131]

[0132] According to the synthesis method of compound 5 in Example 1, compound 30 was synthesized according to the above route as a yellow solid with a yield of 29.1%. 1 H NMR(400MHz,DMSO-d6)δ9.14(s,2H),8.32–8.29(m,2H),7.10–7.07(m,4H),3 .89(s,6H),3.35–3.30(m,4H),2.98(t,J=4.9Hz,4H),1.24(d,J=2.2Hz,1H). 13 C NMR (400MHz, DMSO-d6) δ162.55,155.51,154.02,153.03,138.31,130.29,130.16,129.32,127.29,114.97,104.45,60.61,56.59,47.11,44.58.

[0133] Comparative Example 6: Synthesis of Compounds 23, 24, 26, and 28

[0134] Compound 26

[0135]

[0136] According to the synthesis method of compound 25, compound II and 4-morpholinophenylboronic acid pinacol ester were used as raw materials to obtain a yellow solid with a yield of 20.1%. 1 H NMR(400MHz,DMSO-d6)δ9.55(s,1H),8.80(s,2H),7.66–7.61(m,2H),6.96(s,2H), 6.94–6.88(m,2H),3.86(s,7H),3.76–3.72(m,4H),3.68(s,3H),3.06–3.01(m,4H). 13 C NMR (400MHz, DMSO-d6) δ159.78,156.19,153.92,146.68,137.42,133.31,131.01,124.01,120.64,116.09,103.51,66.64,60.53,56.52,49.77

[0137] Intermediate series 10 compound II-1

[0138]

[0139] According to the above route, compound I and 2-chloro-5-bromopyrimidine were used as raw materials to prepare series 10 compound II-1. Yellow solid. The yield was 68.9%.

[0140] Compound 24

[0141]

[0142] According to the synthesis method of compound 25, series 10 compound II-1 and 4-morpholinophenylboronic acid pinacol ester were used as raw materials to obtain a yellow solid with a yield of 23.1%. 1HNMR (400MHz, DMSO-d6) δ9.62(s,1H),8.86(s,2H),8.23(t,J=1.8Hz,1H),7.97(dt,J=7.7,1.4Hz,1H),7.92(dt,J=7.9,1.3Hz,1H),7 .64(d,J=4.1Hz,3H),7.62(d,J=2.8Hz,3H),7.44–7.34(m,1H),6.96–6.87(m,2H),3.81–3.69(m,4H),3.10–2.98(m,4H),2.67(s,3H).

[0143] Compound 23

[0144]

[0145] According to the synthesis method of compound 12, 2,5-dichloropyrimidine and 4-4-methylpiperazineaniline are used as raw materials, and the reaction is carried out at 120°C, and the methyl group on piperazine is removed, resulting in disubstitution. Black solid. 1 H NMR (400MHz, DMSO-d6) δ9.62(s,1H),8.47(d,J=6.6Hz,4H),7.58–7.47(m,2H),6.99–6.88(m,2H),3.87(t,J=5.2Hz,4H),3.14(t,J=5.2Hz,4H). 13 CNMR(400MHz,DMSO-d6)δ164.92,159.97,158.95,156.52,146.93,132.87,121.09,119.12,118.06,116.95,49.45,44.07.Exact Mass:401.0922; Found:402.1000

[0146] Compound 28

[0147]

[0148] According to the synthesis method of series 2 intermediate compound III-1, compound 28 was synthesized according to the above route. White solid. The yield was 54.6%. 1 H NMR(600MHz,Chloroform-d)δ9.13(s,2H),7.85(dd,J=8.3,1.4Hz,1H),7.77(d,J=7.9Hz,1H),7.65(d,J=7.0Hz,2H),7.55(t,J=7.5Hz,2H),7.49( t,J=7.4Hz,1H),7.10(dd,J=8.2,1.9Hz,1H),7.07(d,J=1.9Hz,1H),7.02(dd,J=7.9,1.7Hz,1H),6.91(d,J=1.7Hz,1H),3.97(s,3H),3.92(s,3H). 13 C NMR(151MHz,Chloroform-d)δ164.97,158.57,155.00,138.48,137.81,133.88,132. 88,131.65,129.52,129.15,126.97,121.52,121.06,112.68,110.91,56.53,55.83.

[0149] Comparative Example 7, Synthesis of Compounds 15 and 16

[0150] Compound 16

[0151]

[0152] According to the synthesis method of compound 17, series 4 compound II-2 was used as raw material to prepare series 4 compound III-2 (ie compound 16), a yellow solid, with a yield of 12.8%. 1 H NMR(400MHz,DMSOd6)δ9.77(s,1H),8.93(s,2H),8.64–8.58(m,2H),7.79–7.73(m,2H),7.66 –7.58(m,2H),6.97–6.92(m,2H),3.46(d,J=5.6Hz,4H),3.03(t,J=5.2Hz,4H),1.42(s,9H). 13 C NMR(400MHz,DMSO-d6)δ160.66,156.76,154.33,150.49,146.96,142.76,132 .93,121.20,120.63,120.01,117.04,79.43,49.66(2C),28.54.HRMS(ESI)m / z for[M+H] + :Exact Mass:432.2274;Found:433.2365Compound 15

[0153]

[0154] According to the synthesis method of compound 14, series 4 compound III-2 was used as raw material to prepare series 4 compound VI-2 (ie compound 15), a yellow solid, with a yield of 30.5%. 1 HNMR(400MHz,DMSO-d6)δ9.73(s,1H),8.92(s,2H),8.61–8.58(m,2H),7.77–7.73(m,2H),7.61 –7.56(m,2H),6.92–6.85(m,2H),2.98(dd,J=6.3,3.6Hz,4H),2.85–2.79(m,4H),1.23(s,1H). 13 C NMR(400MHz,DMSO-d6)δ160.67,156.70,150.68,147.86,142.56,132.21,121.24,120.55,119.93,116.20,50.62,46.17.HRMS(ESI)m / z for[M+H] + :Exact Mass:332.1749;Found:332.1871

[0155] Comparative Example 8: Synthesis of Compound 31

[0156] Compound 31

[0157]

[0158] According to the synthesis method of 32, compound 31 was prepared as a yellow solid using series 9 compound I and 4-(4-methylpiperazine)aniline as raw materials (the equivalent ratio of the two materials was 1.5:1). The yield was 27.7%. 1 H NMR(400MHz,DMSO-d6)δ8.94(s,4H),7.78–7.75(m,4H),7.54–7.49(m,4H),7.46–7.40(m,2H),7 .09–7.06(m,2H),6.99–6.94(m,2H),3.16(t,J=5.0Hz,4H),2.47(d,J=5.0Hz,4H),2.24(s,3H). 13 C NMR (400MHz, DMSO-d6) δ162.11,156.41,149.76,134.87,134.45,129.69,128.72,128.68,128.21,126.83,116.39,116.17,55.07,48.63.

[0159] The beneficial effects of the present invention are demonstrated by experiments as follows.

[0160] Experimental Example 1: Cell proliferation inhibition experiment

[0161] 1. Experimental Methods

[0162] HCT-116 cells from ScienCell were cultured in a medium containing DMEM and placed in an incubator with a 5% CO2 concentration. After the cells were fully grown in the culture dish, they were passaged at a ratio of 1:2. HCT-116 cells of generations 3-8 were selected for the experiment. The inhibitory effect of the synthesized target compound on the proliferation of HCT-116 cells was measured using the MTT method, and AZ191 was selected as a positive control drug. IC was calculated using SPSS software 50 value.

[0163] 1) Experimental principle

[0164] Thiazolyl blue, referred to as MTT, is a detection reagent that can be used to detect the content of live cells and cytotoxicity. Succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-purple crystalline formazan and deposit it in the cells, while dead cells do not have this function. Dimethyl sulfoxide (DMSO) can dissolve the formazan in cells, and its light absorption value can be measured at a wavelength of 490nm using an enzyme-linked immunosorbent assay, which can indirectly reflect the number of live cells. Within a certain range of cell numbers, the amount of MTT crystals formed is proportional to the number of cells.

[0165] 2) Experimental Materials

[0166] a. Cell line: Human colon cancer cell line (HCT-116)

[0167] b. Reagents and instruments: DMEM medium (HyClone), culture dishes (JETBIOFIL), 96-well plates (CorningCostar), trypsin (Biological Industries, BI); MTT reagent (Biofroxx); cell culture incubator (ThermoFisher); microplate reader (BioTek Instruments, Inc).

[0168] 3) Experimental methods

[0169] a. Cell culture and plating: cells were cultured in DMEM medium. After the cells were fully grown, the medium was discarded, and the cells were washed with PBS for 3 times. An appropriate amount of trypsin was added and the cells were digested in an incubator for 1 min. The digestion was terminated with DMEM medium, and the cells were blown off and placed in a centrifuge tube. The cells were centrifuged at 1200 rpm for 4 min, and the supernatant was discarded. An appropriate amount of fresh DMEM medium was added and the cells were blown off and mixed. The cells were counted with a cell counting plate to obtain the number of tumor cells in the medium.

[0170] b. Cell dosing: dilute the cell-containing medium to a suitable concentration and add it to a 96-well plate, with 180 μl of medium per well. The number of cells in each well is maintained at 1×10 4 Then, the cells were placed in a 5% CO2 incubator for 24 hours. After the cells adhered to the wall, the medium was aspirated and 200 μl of serum-free medium containing blank concentration, 0.25 μM, 0.5 μM, 1 μM, 2 μM, 5 μM, 10 μM and 20 μM concentrations of compound 20 were added. Three parallel control groups were set up for the wells with different concentrations. The 96-well plate after drug addition was transferred to a low oxygen incubator with 1% O2 and 5% CO2 and cultured for 48 hours. After 48 hours, the medium was removed and 50 μl of medium containing 2% MTT was added. The OD value was detected after incubation for 5 hours.

[0171] c. Data processing: IC 50 The value was calculated by SPSS. The anti-proliferation level of the compound on cells was calculated according to the following formula:

[0172] Inhibition rate (%) = (blank control well OD value - drug addition well OD value) / (blank control well OD value - cell-free well OD value) × 100%

[0173] 2. Experimental results

[0174] Table 1. Inhibitory activity of compounds on colon cancer cell proliferation

[0175]

[0176]

[0177] The results are shown in Table 1. It can be seen that compared with AZ191, only compounds 14, 17, and 32 of the present invention have better inhibitory activity on colon cancer cell proliferation.

[0178] The above experimental results show that the compounds of the present invention have broad application prospects in the preparation of drugs for preventing and / or treating tumors.

Claims

1. A compound, a stereoisomer thereof, a deuterated substance thereof or a pharmaceutically acceptable salt thereof, characterized in that: The structure of the compound is shown in Formula I: Where X1 is N or CR a , X2 is N or CR b , X3 is N or CR c , X4 is N or CR d , X5 is N or CR e , R a , R b , R c , R d , R e Each independently selected from hydrogen, C 1-8 Alkyl or C 1-8 Alkoxy; R2 is selected from hydrogen, C 1-8 Alkyl or C 1-8 Alkoxy; R1 is selected from hydrogen, C 1-8 Alkyl, COOR4, COR4, R4 is selected from C 1-8 alkyl.

2. The compound according to claim 1, its stereoisomer, its deuterated substance or its pharmaceutically acceptable salt, characterized in that: The structure of the compound is shown in Formula II: Where X1 is N or CR a , X2 is N or CR b , X3 is N or CR c , X4 is N or CR d , X5 is N or CR e , R a , R b , R c , R d , R e Each independently selected from hydrogen, C 1-4 Alkyl or C 1-4 Alkoxy; R2 is selected from hydrogen, C 1-4 Alkyl or C 1-4 Alkoxy; R1 is selected from hydrogen, C 1-4 Alkyl, COOR4, COR4, R4 is selected from C 1-4 alkyl.

3. The compound according to claim 2, its stereoisomer, its deuterated substance or its pharmaceutically acceptable salt, characterized in that: The structure of the compound is shown in Formula III: Wherein R1 is selected from hydrogen, C 1-4 Alkyl, COOR4, COR4, R4 is selected from C 1-4 alkyl.

4. The compound according to claim 3, its stereoisomer, its deuterated substance or its pharmaceutically acceptable salt, characterized in that: The R4 is tert-butyl.

5. The compound according to claim 1, its stereoisomer, its deuterated substance or its pharmaceutically acceptable salt, characterized in that: The compound is selected from one of the following compounds:

6. A pharmaceutical composition, characterized in that The pharmaceutical composition is a preparation prepared with the compound according to any one of claims 1 to 5, its stereoisomer, its deuterated product or its pharmaceutically acceptable salt as the active ingredient and pharmaceutically acceptable excipients.

7. The pharmaceutical composition according to claim 6, characterized in that The preparation is an oral preparation or an injection preparation.

8. Use of the compound according to any one of claims 1 to 5, its stereoisomer, its deuterated product or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating tumors.

9. The use according to claim 8, characterized in that The tumor is colon cancer, lung cancer, breast cancer, stomach cancer, liver cancer, esophageal cancer, cervical cancer, prostate cancer or lymphoma.

10. The use according to claim 9, characterized in that The tumor is colon cancer.

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