Phenylureido quinazolines, their preparation and use in the preparation of medicaments for the prevention or treatment of liver cancer

Abstract

The application discloses a phenyl urea quinazoline compound and preparation and application thereof in preparation of a medicine for preventing or treating liver cancer. The phenyl urea quinazoline compound has a structure as shown in formula (IA) or (IB). In formula (IA), R is H, 4-methoxy, 2,5-difluoro or 3-methyl; and in formula (IB), R is H, 3-trifluoromethyl-4-chloro, 4-chloro or 3-methoxy. The phenyl urea quinazoline compound provided by the application has a good inhibiting effect on human liver cancer cells; the preparation method of the compound is simple and easy to operate, raw materials are easy to obtain, the production cost is low, and the compound is suitable for industrial application.

Description

Technical Field

[0001] This invention relates to a quinazoline compound and its application, particularly to a phenylurea-quinazoline compound and its preparation method, as well as the application of said compound or a pharmaceutically acceptable salt thereof in the preparation of drugs for the prevention or treatment of liver cancer. Background Technology

[0002] Quinazoline compounds possess many beneficial biological activities and have wide applications in the pharmaceutical field. In particular, some quinazoline derivatives with specific structures exhibit significant antiviral, antibacterial, and antitumor activities. Several quinazoline compounds have already been marketed as antitumor drugs. For example, gefitinib and erlotinib, used to treat lung cancer, and lapatinib, used to treat breast cancer, are all quinazoline compounds. Novel quinazoline compounds and their biological activities are also frequently reported in the literature (see Y.-Y.Ke, H.-Y.Shiao, YCHsu, C.-Y.Chu, W.-C.Wang, Y.-C.Lee, W.-H.Lin, C.-H.Chen, JTAHsu, C.-W.Chang, C.-W.Lin, T.-K.Yeh, Y.-S.Chao, MSCoumar, H.-P.Hsieh, ChemMedChem 2013, 8, 136-148; A.Garofalo, A.Farce, S.Ravez, A.Lemoine, P.Six, P.Chavatte, L.Goossens, P.Depreux, J.Med.Chem. 2012, 55, 1189-1204). However, most quinazoline compounds do not possess antitumor activity. Summary of the Invention

[0003] The purpose of this invention is to provide a novel quinazoline compound with anticancer activity—phenylurea quinazoline compound, its preparation method, and its application. This compound has a good inhibitory effect on human liver cancer cells at a certain dose. Moreover, the preparation method of this compound is simple, easy to operate, the raw materials are readily available, and the production cost is low, making it suitable for industrial application.

[0004] To achieve the above-mentioned objectives, the present invention adopts the following technical solution:

[0005] In a first aspect, the present invention provides a phenylurea-quinazoline compound represented by formula (IA) or (IB).

[0006]

[0007] In formula (IA), R is H, 4-methoxy, 2,5-difluoro, or 3-methyl, with the specific structure as follows:

[0008]

[0009] In formula (ⅠB), R is H, 3-trifluoromethyl-4-chloro, 4-chloro, or 3-methoxy, with the following specific structure:

[0010]

[0011] In a second aspect, the present invention provides a method for preparing phenylurea-quinazoline compounds, the method comprising: (1) adding the compound shown in formula (II) to organic solvent A, stirring to dissolve, and adding dropwise a mixture containing the compound shown in formula (IIIA) or (IIIB), catalyst B and organic solvent A under stirring conditions at room temperature. After the addition is complete, the reaction solution is heated to reflux and stirred for 0.5 to 12 hours. The reaction solution is then separated and purified to obtain the phenylurea-quinazoline compound shown in formula (IA) or (IB).

[0012] The organic solvent A is one of the following: dichloromethane, ethanol, isopropanol, or toluene;

[0013] Catalyst B is one of the following: triethylamine, 4-dimethylaminopyridine (DMAP), pyridine, or sodium hydroxide;

[0014]

[0015] In equation (IIIA), the definition of R is the same as in equation (IA); in equation (IIIB), the definition of R is the same as in equation (IB).

[0016] Furthermore, the molar ratio of the compound shown in formula (II), formula (IIIA) or (IIIB) to the catalyst B is 1:0.8 to 1.2:0.1 to 1.

[0017] Furthermore, the amount of organic solvent A used is sufficient to dissolve the compound shown in formula (II), the compound shown in formula (IIIA) or (IIIB), and the catalyst B. Preferably, the total volume of organic solvent A used is 10 to 30 mL / mmol based on the amount of substance of the compound shown in formula (II), wherein the volume of organic solvent A used to dissolve the compound shown in formula (II) is 4 to 10 mL / mmol based on the amount of substance of the compound shown in formula (II), and the volume of organic solvent A used to dissolve the compound shown in formula (IIIA) or (IIIB) and the catalyst B is 6 to 20 mL / mmol based on the amount of substance of the compound shown in formula (II).

[0018] Furthermore, the reaction process was monitored by TLC (using a mixture of petroleum ether and ethyl acetate in a volume ratio of 0.5 to 10:1) to determine the reaction endpoint. The reaction time was generally 0.5 to 12 hours.

[0019] Furthermore, the separation and purification described in step (1) of this invention adopts the following steps: after the reaction is completed, the solvent is removed by evaporation, and the residue is subjected to column chromatography to obtain phenylurea-quinazoline compounds.

[0020] Furthermore, the column chromatography operation steps in step (1) of this invention are as follows: Take the residue after solvent removal into a single-necked bottle, add organic solvent C to dissolve it, and obtain a solution. Then, add 1 to 4 times the mass of the residue to the solution to column chromatography silica gel (preferably 300 to 400 mesh coarse pore (ZCX.II) type column chromatography silica gel). After mixing, remove the solvent to obtain a dry mixture of residue and silica gel. Load the mixture onto a column and elute it with a petroleum ether and ethyl acetate mixed solution with a volume ratio of 0.5 to 10:1. Perform elution and TLC monitoring (developing solvent is a petroleum ether and ethyl acetate mixed solution with a volume ratio of 0.5 to 10:1). Collect the eluent containing phenylureaquinazoline compounds according to the TLC detection. Concentrate and dry the eluent to obtain phenylureaquinazoline compounds. The organic solvent C is one of the following: petroleum ether, dichloromethane, chloroform, or ethyl acetate. The amount of organic solvent C used is sufficient to dissolve the residue.

[0021] The organic solvents A and C mentioned in this invention are both organic solvents. They are named to facilitate the distinction between different organic solvents used in different steps, and the letters themselves have no meaning.

[0022] Thirdly, the present invention also relates to the use of phenylureaquinazoline compounds or pharmaceutically acceptable salts thereof in the preparation of medicaments for the prevention or treatment of liver cancer.

[0023] Preferably, the drug for preventing or treating liver cancer is a drug for preventing or treating human liver cancer cells HepG2.

[0024] Preferably, the drug for preventing or treating liver cancer is a drug for preventing or treating human liver cancer cells Huh7; in the structural formula (IA) of the phenylurea quinazoline compound, R is 2,5-difluoro (i.e., compound (IA-6)); in formula (IB), R is 3-trifluoromethyl-4-chloro or 3-methoxy (i.e., compound (IB-2) or (IB-4)).

[0025] In particular, the compounds (IB-2) and (IB-3) provided by this invention also have good inhibitory effects on breast cancer.

[0026] The term "pharmaceutically acceptable" in this application means that the compound is chemically and / or toxicologically compatible with other components constituting the formulation and / or with humans or mammals for the prevention or treatment of diseases or conditions.

[0027] The term "pharmaceutically acceptable salt" refers to a relatively non-toxic addition salt of an inorganic or organic acid of the compounds of this invention. For example, see SMBerge et al., "Pharmaceutical Salts," J. Pharm. Sci. 1977, 66, 1-19. Inorganic acids include, for example, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, or nitric acid; organic acids include, formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, hexanoic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)-benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, bamoic acid, pectinic acid, 3-phenylpropionic acid, picric acid, etc. Pteropenic acid, 2-hydroxyethanesulfonic acid, itaconic acid, aminosulfonic acid, trifluoromethanesulfonic acid, dodecyl sulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucoheponic acid, glycerophosphate, aspartic acid, sulfosalicylic acid, etc.

[0028] The beneficial effects of the present invention are mainly reflected in: (1) The phenylurea quinazoline compound (Ⅰ) described in the present invention has good anti-liver cancer activity and is expected to be used in the preparation of drugs for the prevention or treatment of liver cancer; (2) The preparation method of the phenylurea quinazoline compound provided by the present invention is simple and easy to operate, the raw materials are readily available, and the production cost is low, making it suitable for practical use. Detailed Implementation

[0029] The present invention will be further described in conjunction with specific embodiments. The following embodiments are illustrative of the present invention and are not intended to limit the present invention in any way.

[0030] Unless otherwise specified in the embodiments of this invention, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained through conventional technical means or commercially available.

[0031] Compound (II) was prepared according to the methods described in the literature (Journal of Medicinal Chemistry, 2022, 65(10), 7246-7261; Bioorganic Chemistry, 2021, 117, 105407).

[0032] The specific structure of compound (ⅢA) is as follows:

[0033]

[0034] The specific structure of compound (ⅢB) is as follows:

[0035]

[0036] The specific structure of compound (IB) is as follows:

[0037]

[0038] Example 1: Preparation of compound (ⅢA-1)

[0039] 1.12 g (12.0 mmol) of aniline and 1.22 g (10.0 mmol) of DMAP were added to 30 mL of dichloromethane and stirred. Under 0–5 °C, 30 mL of a dichloromethane solution containing 1.64 g (10.0 mmol) of 4-nitrophenyl isocyanate was added dropwise. After the addition was complete, the reaction was carried out at room temperature for 20 h. The mixture was washed with water (25 mL × 3), the organic phase was separated, dried, filtered, and the solvent was removed by vacuum evaporation. The residue was recrystallized from ethanol to give 1-(4-nitrophenyl)-3-phenylurea, with a yield of 71%.

[0040] 1.29 g (5.0 mmol) of 1-(4-nitrophenyl)-3-phenylurea was added to 30 mL of ethanol, followed by 0.20 g of Pd / C (5%). The reaction was carried out under stirring at room temperature and hydrogen gas was introduced at atmospheric pressure. The reaction was monitored by TLC (using a 1:1 mixture of petroleum ether and ethyl acetate as the developing solvent) until the reaction was complete. The mixture was filtered, the solvent was removed by vacuum evaporation, and the residue was recrystallized from ethanol to give product (ⅢA-1) with a yield of 83%.

[0041] Preparation of compounds (ⅢA-2) to (ⅢA-9): The corresponding compounds (ⅢA-2) to (ⅢA-9) were prepared by replacing aniline with the corresponding substituted aniline, following the method in Example 1. Further details will not be provided here.

[0042] Example 2: Preparation of compound (ⅢB-1)

[0043] 1.29 g (12.0 mmol) of benzoylamine and 1.22 g (10.0 mmol) of DMAP were added to 30 mL of dichloromethane and stirred. Under 0–5 °C, 30 mL of a dichloromethane solution containing 1.64 g (10.0 mmol) of 4-nitrophenyl isocyanate was added dropwise. After the addition was complete, the reaction was carried out at room temperature for 22 h. The mixture was washed with water (25 mL × 3), the organic phase was separated, dried, filtered, and the solvent was removed by vacuum evaporation. The residue was recrystallized from ethanol to give 1-(4-nitrophenyl)-3-benzylurea, with a yield of 65%.

[0044] 1.36 g (5.0 mmol) of 1-(4-nitrophenyl)-3-benzylurea was added to 30 mL of ethanol, followed by 0.20 g of Pd / C (5%). The reaction was carried out under stirring at room temperature and hydrogen gas was introduced at atmospheric pressure. The reaction was monitored by TLC (using a 1:1 mixture of petroleum ether and ethyl acetate as the developing solvent) until the reaction was complete. The mixture was filtered, the solvent was removed by vacuum evaporation, and the residue was recrystallized from ethanol to give product (ⅢB-1) with a yield of 78%.

[0045] Preparation of compounds (ⅢB-2) to (ⅢB-5): The corresponding compounds (ⅢB-2) to (ⅢB-5) were prepared by replacing benzylamine with the corresponding substituted benzylamine, referring to the method in Example 2. The details will not be repeated here.

[0046] Example 3: Preparation of compound (IA-1)

[0047] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.455 g (2.0 mmol) of compound (ⅢA-1) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture in an oil bath to reflux for 12 hours (the reaction process is monitored by TLC, with a 1:2 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and then perform column chromatography on the residue. Dissolve the residue in 10 mL of petroleum ether to obtain the solution. Then, add the solution to... Add 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel), mix well, evaporate the solvent, and obtain a dry residue mixed with silica gel. Pack the mixture into a column, and then elute with a 1:2 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. Follow the TLC detection (developing solvent is a 1:2 volume ratio of petroleum ether and ethyl acetate mixture). Collect the eluent containing the compound shown in formula (IA-1) according to the TLC detection. Evaporate the solvent from the collected eluent and dry to obtain a white solid product, namely compound (IA-1), with a yield of 63% (based on the molar amount of 2,4-dichloroquinazoline (II)) and a melting point of 218-220℃. 1HNMR(500MHz,DMSO-d6)δ10.16(s,1H),8.72(s,1H),8.69(s,1H),8.54(d,J=8.1Hz,1H),7.91-7.83(m,1H),7.65(d,J=7.9 Hz,2H),7.63(d,J=8.2Hz,2H),7.51(d,J=8.9Hz,2H),7.46(d,J=7.6Hz,2H),7.28(t,J=7.9Hz,2H),6.97(t,J=7.3Hz,1H). 13 C NMR (125MHz, DMSO-d6) δ155.2,154.6,153.2,152.5,151.3,137.1,133.6,132.3,122.1,119.4,119.1,119.0,118.8,118.4,118.3,116.9,116.7.

[0048] Example 4: Preparation of compound (IA-1)

[0049] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 8 mL of toluene. Under magnetic stirring at room temperature, add dropwise 12 mL of toluene solution containing 0.545 g (2.4 mmol) of compound (ⅢA-1) and 0.008 g (0.2 mmol) of sodium hydroxide. After the addition is complete, heat the reaction mixture to reflux in an oil bath and reflux for 0.5 hours (the reaction process is monitored by TLC, with a 10:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and perform column chromatography on the residue. Dissolve the residue in 10 mL of dichloromethane to obtain the solution. Then add... 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel) was mixed, and the solvent was evaporated to obtain a dry residue mixed with silica gel. The mixture was packed into a column, and then eluted with a 10:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. TLC was used for monitoring (developing solvent was a 10:1 volume ratio of petroleum ether and ethyl acetate mixture). The eluent containing the compound shown in formula (IA-1) was collected according to the TLC detection. The collected eluent was evaporated to remove the solvent and dried to obtain a white solid product, namely compound (IA-1), with a yield of 45% (based on the molar amount of 2,4-dichloroquinazoline (II)) and a melting point of 218-220℃. 1 H NMR and 13 C NMR is the same as in Example 1.

[0050] Example 5: Preparation of compound (IA-1)

[0051] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of ethanol. Under magnetic stirring at room temperature, add dropwise a 40 mL ethanol solution containing 0.364 g (1.6 mmol) of compound (ⅢA-1) and 0.202 g (2.0 mmol) of triethylamine. After the addition is complete, heat the reaction solution to reflux in an oil bath and reflux for 6 hours (the reaction process is monitored by TLC, with a 1:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction solution, and perform column chromatography on the residue. Dissolve the residue in 10 mL of chloroform to obtain the solution. Then add... 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel) was mixed, and the solvent was evaporated to obtain a dry residue mixed with silica gel. The mixture was packed into a column, and then eluted with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. TLC was used for monitoring (developing solvent was a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). The eluent containing the compound represented by formula (IA-1) was collected according to the TLC detection. The collected eluent was evaporated to remove the solvent and dried to obtain a white solid product, namely compound (IA-1), with a yield of 55% (based on the molar amount of compound (IIIA-1)) and a melting point of 218-220℃. 1 H NMR and 13 C NMR is the same as in Example 1.

[0052] Example 6: Preparation of compound (IA-1)

[0053] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 10 mL of isopropanol. Under magnetic stirring at room temperature, add dropwise a 15 mL isopropanol solution containing 0.455 g (2.0 mmol) of compound (ⅢA-1) and 0.079 g (1.0 mmol) of pyridine. After the addition is complete, heat the reaction solution to reflux in an oil bath and reflux for 4 hours (the reaction process is monitored by TLC, with a 5:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction solution, and perform column chromatography on the residue. Dissolve the residue in 10 mL of ethyl acetate to obtain the solution. Then add... 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel) was mixed, and the solvent was evaporated to obtain a dry residue mixed with silica gel. The mixture was packed into a column, and then eluted with a 5:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. TLC was used for monitoring (developing solvent was a 5:1 volume ratio of petroleum ether and ethyl acetate mixture). The eluent containing the compound shown in formula (IA-1) was collected according to the TLC detection. The collected eluent was evaporated to remove the solvent and dried to obtain a white solid product, namely compound (IA-1), with a yield of 42% (based on the molar amount of 2,4-dichloroquinazoline (II)) and a melting point of 218-220℃. 1 H NMR and 13 C NMR is the same as in Example 1.

[0054] Example 7: Preparation of compound (IA-2)

[0055] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.511 g (2.0 mmol) of compound (ⅢA-2) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture in an oil bath to reflux for 10 hours (the reaction process is monitored by TLC, with a 1:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and then perform column chromatography on the residue. Dissolve the residue in 10 mL of petroleum ether to obtain the solution. Then, add the solution to... Add 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel), mix well, evaporate the solvent, and obtain a dry residue mixed with silica gel. Pack the mixture into a column, and then elute with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. Detect by TLC (developing solvent is a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). Collect the eluent containing the compound shown in formula (IA-2) according to the TLC detection. Evaporate the solvent from the collected eluent and dry to obtain a white solid product, namely compound (IA-2), with a yield of 61% (based on the molar amount of 2,4-dichloroquinazoline (II)) and a melting point of 232-234 °C. 1 HNMR(500MHz,DMSO-d6)δ10.19(s,1H),9.11(s,1H),8.56(d,J=1.9Hz,1H) ,8.12(td,J=9.3,6.2Hz,1H),7.96(s,1H),7.88-7.84(m,1H),7.71(dd,J=8 .4,1.2Hz,1H),7.69-7.64(m,2H),7.52(d,J=8.9Hz,2H),7.33-7.29(m,1H) ,7.27-7.24(m,1H),7.07(dd,J=8.6,2.9Hz,1H),2.90(s,3H),2.79(s,3H).

[0056] Example 8: Preparation of compound (IA-3)

[0057] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.515 g (2.0 mmol) of compound (ⅢA-3) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture in an oil bath to reflux for 9 hours (the reaction process is monitored by TLC using a 1:1 mixture of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and then perform column chromatography on the residue. Dissolve the residue in 10 mL of petroleum ether to obtain the solution. Then, add the solution to... Add 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel), mix well, evaporate the solvent, and obtain a dry residue mixed with silica gel. Pack the mixture into a column, and then elute with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. Follow the TLC detection (developing solvent is a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). Collect the eluent containing the compound shown in formula (IA-3) according to the TLC detection. Evaporate the solvent from the collected eluent and dry to obtain a white solid product, namely compound (IA-3), with a yield of 68% (based on the molar amount of 2,4-dichloroquinazoline (II)) and a melting point of 265-267℃. 1 HNMR(500MHz,DMSO-d6)δ10.15(s,1H),8.68(s,1H),8.54(s,1H),7.86-7.82(m,1H),7.69(dd,J=8.4,1.2Hz,1H ),7.67-7.58(m,3H),7.55-7.44(m,2H),7.42-7.28(m,2H),7.22-7.07(m,1H),6.96-6.76(m,2H),3.71(s,3H); 13 C NMR(125MHz,DMSO-d6)δ163.0,159.6,156.6,154.6,153.0,150.8,150.5,1 41.0,137.3,133.0,132.0,124.1,122.5,120.1,118.3,114.2,113.9,55.4.

[0058] Example 9: Preparation of compound (IA-4)

[0059] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.592 g (2.0 mmol) of compound (ⅢA-4) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture in an oil bath to reflux for 10 hours (the reaction process is monitored by TLC, with a 1:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and then perform column chromatography on the residue. Dissolve the residue in 7 mL of petroleum ether to obtain the solution. Then, add the solution to... Add 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel), mix well, evaporate the solvent, and obtain a dry residue mixed with silica gel. Pack the mixture into a column, and then elute with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. Detect by TLC (developing solvent is a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). Collect the eluent containing the compound shown in formula (IA-4) according to the TLC detection. Evaporate the solvent from the collected eluent and dry to obtain a white solid product, namely compound (IA-4), with a yield of 51% (based on the molar amount of 2,4-dichloroquinazoline (II)) and a melting point of 188-189 °C. 1 HNMR(500MHz,DMSO-d6)δ10.18(s,1H),9.05(s,1H),8.89(s,1H),8.54-8.51(m,1H),7.96 -7.81(m,2H),7.75-7.60(m,4H),7.52(dd,J=8.8,4.3Hz,3H),7.34(dd,J=8.8,2.5Hz,1H); 13 CNMR(125MHz,DMSO-d6)δ159.4,156.4,152.4,150.7,140.1,136.3,134.0,132.5 ,131.1,130.62,126.8,126.5,123.9,123.4,123.0,119.2,118.6,118.2,113.8.

[0060] Example 10: Preparation of compound (IA-5)

[0061] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.511 g (2.0 mmol) of compound (ⅢA-5) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture in an oil bath to reflux for 8 hours (the reaction process is monitored by TLC, with a 1:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and then perform column chromatography on the residue. Dissolve the residue in 10 mL of petroleum ether to obtain the solution. Then, add the solution to... Add 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel), mix well, evaporate the solvent, and obtain a dry residue mixed with silica gel. Pack the mixture into a column, and then elute with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. Follow the TLC detection (the developing solvent is a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). Collect the eluent containing the compound shown in formula (IA-5) according to the TLC detection. Evaporate the solvent from the collected eluent and dry to obtain a white solid product, namely compound (IA-5), with a yield of 67% (based on the molar amount of 2,4-dichloroquinazoline (II)) and a melting point of 204-206 °C. 1 HNMR(500MHz,DMSO-d6)δ10.15(s,1H),8.69(s,1H),8.54(d,J=7.3Hz,2H),7.95-7.78(m,2H),7.73-7 .67(m,1H),7.65(s,1H),7.66-7.60(m,3H),7.51(d,J=7.4Hz,2H),7.50(d,J=7.4Hz,1H),2.24(s,6H); 13 C NMR(125MHz,DMSO-d6)δ159.6,156.6,152.7,150.9,139.8,137.9,137.0,1 34.1,132.2,127.1,127.0,126.7,124.1,123.6,118.4,116.1,113.9,21.3.

[0062] Example 11: Preparation of compound (IA-6)

[0063] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.526 g (2.0 mmol) of compound (ⅢA-6) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture to reflux in an oil bath and reflux for 11 hours (the reaction process is monitored by TLC, with a 1:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and then perform column chromatography on the residue. Dissolve the residue in 10 mL of petroleum ether to obtain the solution. Then, add the solution to... Add 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel), mix well, evaporate the solvent, and obtain a dry residue mixed with silica gel. Pack the mixture into a column, and then elute with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. Follow the TLC detection (developing solvent is a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). Collect the eluent containing the compound shown in formula (IA-6) according to the TLC detection. Evaporate the solvent from the collected eluent and dry to obtain a white solid product, namely compound (IA-6), with a yield of 67% (based on the molar amount of 2,4-dichloroquinazoline (II)) and a melting point of 238-239 °C. 1 HNMR(500MHz,DMSO-d6)δ9.91(s,1H),8.90(s,1H),8.79(s,1H),8.36(d,J=6.4Hz,1H),7.82(dd,J=6.7,1.8Hz,1H),7.73( d,J=8.3Hz,2H),7.50(d,J=6.0Hz,1H),7.49(s,1H),7.47(s,1H),7.32(td,J=6.0,3.8Hz,2H),7.17(td,J=8.4,1.7Hz,2H), 13 C NMR(125MHz,DMSO-d6)δ162.1,159.2,156.2,152.2,150.6,140.7,136.2,133.8 ,132.2,126.8,126.7,126.6,126.3,123.8,123.26,118.1,115.1,113.6,99.4.

[0064] Example 12: Preparation of compound (IA-7)

[0065] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.523 g (2.0 mmol) of compound (ⅢA-7) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture in an oil bath to reflux for 6 hours (the reaction process is monitored by TLC, with a 1:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and then perform column chromatography on the residue. Dissolve the residue in 10 mL of petroleum ether to obtain the solution. Then, add the solution to... Add 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel), mix well, evaporate the solvent, and obtain a dry residue mixed with silica gel. Pack the mixture into a column, and then elute with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. Detect by TLC (developing solvent is a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). Collect the eluent containing the compound shown in formula (IA-7) according to the TLC detection. Evaporate the solvent from the collected eluent and dry to obtain a white solid product, namely compound (IA-7), with a yield of 59% (based on the molar amount of 2,4-dichloroquinazoline (II)) and a melting point of 198-199 °C. 1 HNMR(500MHz,DMSO-d6)δ10.81(s,1H),9.66(d,J=6.4Hz,1H),8.83(s,1H),8.77(s,1H),8.57(dd,J=8.2,2.4Hz,1H),7.8 5(d,J=8.2Hz,1H),7.66(d,J=8.9Hz,2H),7.56-7.53(m,2H),7.47-7.44(m,2H),7.31-7.27(m,2H),6.98(d,J=7.4Hz,1H). 13 C NMR (125MHz, DMSO-d6) δ155.2,154.6,154.4,152.4,141.3,135.2,133.6,133.1,132.3,130.3,124.0,122.1,121.3,118.8,117.5,116.5,100.2.

[0066] Example 13: Preparation of compound (IA-8)

[0067] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.483 g (2.0 mmol) of compound (ⅢA-8) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture in an oil bath to reflux for 5 hours (the reaction process is monitored by TLC, with a 1:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and then perform column chromatography on the residue. Dissolve the residue in 10 mL of petroleum ether to obtain the solution. Then, add the solution to... Add 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel), mix well, evaporate the solvent, and obtain a dry residue mixed with silica gel. Pack the mixture into a column, and then elute with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. Follow the TLC detection (developing solvent is a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). Collect the eluent containing the compound shown in formula (IA-8) according to the TLC detection. Evaporate the solvent from the collected eluent and dry to obtain a white solid product, namely compound (IA-8), with a yield of 55% (based on the molar amount of 2,4-dichloroquinazoline (II)) and a melting point of 182-184 °C. 1 HNMR(500MHz,DMSO-d6)δ10.16(s,1H),8.71(s,1H),8.62(s,1H),8.52(d,J=7.9Hz 1H),7.89-7.85(m,1H),7.70(d,J=7.5Hz 1H),7.65(m,3H),7.53-7.50(m,2H),7.32(s,1H),7.25(d,J=8.35Hz 1H),7.18(m,1H),6.80(d,J=7.35Hz 1H),2.29(s,3H).

[0068] Example 14: Preparation of compound (IA-9)

[0069] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.515 g (2.0 mmol) of compound (ⅢA-9) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture in an oil bath to reflux for 9 hours (the reaction process is monitored by TLC, with a 1:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and then perform column chromatography on the residue. Dissolve the residue in 10 mL of petroleum ether to obtain the solution. Then, add the solution to... Add 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel), mix well, evaporate the solvent, and obtain a dry residue mixed with silica gel. Pack the mixture into a column, and then elute with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. Follow the TLC detection (developing solvent is a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). Collect the eluent containing the compound shown in formula (IA-9) according to the TLC detection. Evaporate the solvent from the collected eluent and dry to obtain a white solid product, namely compound (IA-9), with a yield of 51% (based on the molar amount of 2,4-dichloroquinazoline (II)) and a melting point of 228-230℃. 1 HNMR (500MHz, DMSO-d6) δ10.17(s,1H),8.72(s,2H),8.55(d,J=8.3Hz,1H),7.88(t,J=7.7Hz,1H),7.71(d,J=8.3Hz,1H),7.65(dd,J=8 .4,2.7Hz,3H),7.51(dd,J=6.7,2.2Hz,2H),7.19-7.14(m,2H),6.94(dd,J=8.1,1.9Hz,1H),6.56(dd,J=8.2,2.2Hz,1H),3.74(s,3H).

[0070] Example 15: Preparation of compound (IB-1)

[0071] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.483 g (2.0 mmol) of compound (ⅢB-1) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture in an oil bath to reflux for 12 hours (the reaction process is monitored by TLC, with a 1:2 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and perform column chromatography on the residue. Dissolve the residue in 10 mL of petroleum ether to obtain the solution. Then, add the solution to... Add 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel), mix well, evaporate the solvent, and obtain a dry residue mixed with silica gel. Pack the mixture into a column, and then elute with a 1:2 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. Follow the TLC detection (developing solvent is a 1:2 volume ratio of petroleum ether and ethyl acetate mixture). Collect the eluent containing the compound shown in formula (ⅠB-1) according to the TLC detection. Evaporate the solvent from the collected eluent and dry to obtain a white solid product, namely compound (ⅠB-1), with a yield of 70% (based on the molar amount of 2,4-dichloroquinazoline (Ⅱ), melting point 189-191℃. 1 HNMR(500MHz,DMSO-d6)δ10.19(s,1H),10.10(s,1H),8.55(d,J=8.0Hz,1H),7.88-7.84(m,2H),7.71(d,J=7.9Hz,2H),7 .68(d,J=5.7Hz,3H),7.64-7.61(m,2H),7.16-6.12(m,2H),6.74(dd,J=6.3,2.8Hz,1H),6.61-6.57(m,1H),3.90(s,2H).

[0072] Example 16: Preparation of compound (IB-1)

[0073] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 8 mL of toluene. Under magnetic stirring at room temperature, add dropwise 12 mL of toluene solution containing 0.579 g (2.4 mmol) of compound (ⅢB-1) and 0.008 g (0.2 mmol) of sodium hydroxide. After the addition is complete, heat the reaction mixture to reflux in an oil bath and reflux for 0.5 hours (the reaction process is monitored by TLC, with a 10:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and perform column chromatography on the residue. Dissolve the residue in 10 mL of dichloromethane to obtain the solution. Then add... 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel) was mixed, and the solvent was evaporated to obtain a dry residue mixed with silica gel. The mixture was packed into a column, and then eluted with a 10:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. TLC was used for monitoring (developing solvent was a 10:1 volume ratio of petroleum ether and ethyl acetate mixture). The eluent containing the compound shown in formula (ⅠB-1) was collected according to the TLC detection. The collected eluent was evaporated to remove the solvent and dried to obtain a white solid product, namely compound (ⅠB-1), with a yield of 56% (based on the molar amount of 2,4-dichloroquinazoline (Ⅱ)) and a melting point of 189-191℃. 1 HNMR is the same as in Example 1.

[0074] Example 17: Preparation of compound (IB-1)

[0075] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of ethanol. Under magnetic stirring at room temperature, add dropwise a 40 mL ethanol solution containing 0.386 g (1.6 mmol) of compound (ⅢB-1) and 0.202 g (2.0 mmol) of triethylamine. After the addition is complete, heat the reaction solution to reflux in an oil bath and reflux for 6 hours (the reaction process is monitored by TLC, with a 1:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction solution, and perform column chromatography on the residue. Dissolve the residue in 10 mL of chloroform to obtain the solution. Then add... 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel) was mixed, and the solvent was evaporated to obtain a dry residue mixed with silica gel. The mixture was packed into a column, and then eluted with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. TLC was used for monitoring (developing solvent was a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). The eluent containing the compound represented by formula (ⅠB-1) was collected according to the TLC detection. The collected eluent was evaporated to remove the solvent and dried to obtain a white solid product, namely compound (ⅠB-1), with a yield of 67% (based on the molar amount of compound (ⅢB-1)) and a melting point of 189-191 °C.1 H NMR is the same as in Example 1.

[0076] Example 18: Preparation of compound (IB-1)

[0077] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 10 mL of isopropanol. Under magnetic stirring at room temperature, add dropwise a 15 mL isopropanol solution containing 0.483 g (2.0 mmol) of compound (ⅢB-1) and 0.079 g (1.0 mmol) of pyridine. After the addition is complete, heat the reaction solution to reflux in an oil bath and reflux for 4 hours (the reaction process is monitored by TLC, with a 5:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction solution, and perform column chromatography on the residue. Dissolve the residue in 10 mL of ethyl acetate to obtain the solution. Then add... 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel) was mixed, and the solvent was evaporated to obtain a dry residue mixed with silica gel. The mixture was packed into a column, and then eluted with a 5:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. The eluent was monitored by TLC (the developing solvent was a 5:1 volume ratio of petroleum ether and ethyl acetate mixture). The eluent containing the compound shown in formula (ⅠB-1) was collected according to the TLC detection. The collected eluent was evaporated to remove the solvent and dried to obtain a white solid product, namely compound (ⅠB-1), with a yield of 54% (based on the molar amount of 2,4-dichloroquinazoline (Ⅱ)) and a melting point of 189-191℃. 1 H NMR is the same as in Example 1.

[0078] Example 19: Preparation of compound (IB-2)

[0079] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.687 g (2.0 mmol) of compound (ⅢB-2) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture in an oil bath to reflux for 11 hours (the reaction process is monitored by TLC, with a 1:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and then perform column chromatography on the residue. Dissolve the residue in 10 mL of petroleum ether to obtain the solution. Then, add the solution to... Add 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel), mix well, evaporate the solvent, and obtain a dry residue mixed with silica gel. Pack the mixture into a column, and then elute with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. Follow the TLC detection (the developing solvent is a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). Collect the eluent containing the compound shown in formula (ⅠB-2) according to the TLC detection. Evaporate the solvent from the collected eluent and dry to obtain a white solid product, namely compound (ⅠB-2), with a yield of 55% (based on the molar amount of 2,4-dichloroquinazoline (Ⅱ)) and a melting point of 267-269 °C. 1 HNMR(500MHz,DMSO-d6)δ10.18(d,J=8.8Hz,2H),8.55(dd,J=8.4,1.3Hz,1H),7.89-7.85(m,1H),7.77-7.56 (m,6H),7.39(d,J=8.8Hz,1H),7.07(d,J=6.8Hz,1H),6.85(dd,J=8.8,2.9Hz,1H),6.73(s,1H),3.99(s,2H); 13 C NMR (125MHz, DMSO-d6) δ168.4,159.6,156.5,151.0,147.9,135.8,134.2,133.7,1 32.1,127.0,126.7,123.8,123.6,119.6,116.5,116.0,113.9,111.3,46.8,35.9.

[0080] Example 20: Preparation of compound (IB-3)

[0081] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.551 g (2.0 mmol) of compound (ⅢB-3) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture to reflux in an oil bath and reflux for 10 hours (the reaction process is monitored by TLC, with a 1:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and perform column chromatography on the residue. Dissolve the residue in 10 mL of petroleum ether to obtain the solution. Then, add the solution to... Add 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel), mix well, evaporate the solvent, and obtain a dry residue mixed with silica gel. Pack the mixture into a column, and then elute with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. Follow the TLC detection (the developing solvent is a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). Collect the eluent containing the compound shown in formula (ⅠB-3) according to the TLC detection. Evaporate the solvent from the collected eluent and dry to obtain a white solid product, namely compound (ⅠB-3), with a yield of 67% (based on the molar amount of 2,4-dichloroquinazoline (Ⅱ)) and a melting point of 271-273 °C. 1 HNMR(500MHz,DMSO-d6)δ10.11(s,1H),8.53(d,J=7.3Hz,2H),7.86(m,1H),7.69(dd,J=8.4,1.2Hz,1H),7.62(t,J=6.3Hz,1H ),7.59-7.52(m,2H),7.48-7.40(m,2H),7.40-7.34(m,2H),7.33-7.21(m,2H),6.13(t,J=5.7Hz,1H),2.76(t,J=7.0Hz,2H); 13 C NMR(125MHz,DMSO-d6)δ159.6,156.7,155.4,151.0,138.8,137.8,134.1,1 31.5,130.9,130.8,128.4,127.0,126.6,124.1,123.5,117.9,113.9,35.3.

[0082] Example 21: Preparation of compound (IB-4)

[0083] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.542 g (2.0 mmol) of compound (ⅢB-4) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture in an oil bath to reflux for 8 hours (the reaction process is monitored by TLC using a 1:1 mixture of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and then perform column chromatography on the residue. Dissolve the residue in 7 mL of petroleum ether to obtain the solution. Then, add the solution to the solution... 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel) was added to the column, mixed well, and the solvent was evaporated to obtain a dry residue mixed with silica gel. The mixture was packed into a column, and then eluted with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. TLC was used for monitoring (the developing solvent was a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). The eluent containing the compound shown in formula (IB-4) was collected according to the TLC detection. The collected eluent was evaporated to remove the solvent and dried to obtain a white solid product, namely compound (IB-4), with a yield of 62% (based on the molar amount of 2,4-dichloroquinazoline (II)) and a melting point >300℃. 1 H NMR(500MHz,DMSO-d6)δ10.19(s,1H),10.06(s,1H),8.59-8.49(m,1H),7.88(m,1H),7.79-7.54(m,6H ),7.00(t,J=8.4Hz,1H),6.30-6.10(m,3H),6.03(t,J=6.1Hz,1H),3.87(d,J=6.1Hz,2H),3.67(s,3H); 13 C NMR (125MHz, DMSO-d6) δ169.0,160.2,159.2,156.2,150.7,149.5,135.6,133.8,133 .2,129.5,126.7,126.4,123.5,123.2,119.3,113.6,105.3,101.9,98.2,54.5,47.2.

[0084] Example 22: Preparation of compound (IB-5)

[0085] Dissolve 0.398 g (2.0 mmol) of 2,4-dichloroquinazoline (II) in 20 mL of dichloromethane. Under magnetic stirring at room temperature, add dropwise a 30 mL dichloromethane solution containing 0.511 g (2.0 mmol) of compound (ⅢB-5) and 0.122 g (1.0 mmol) of DMAP. After the addition is complete, heat the reaction mixture in an oil bath to reflux for 10 hours (the reaction process is monitored by TLC, with a 1:1 volume ratio of petroleum ether and ethyl acetate as the developing solvent). Evaporate the solvent from the reaction mixture, and then perform column chromatography on the residue. Dissolve the residue in 10 mL of petroleum ether to obtain the solution. Then, add the solution to... Add 1.5 g of silica gel (300-400 mesh coarse-pore (ZCX.II) type column chromatography silica gel), mix well, evaporate the solvent, and obtain a dry residue mixed with silica gel. Pack the mixture into a column, and then elute with a 1:1 volume ratio of petroleum ether and ethyl acetate mixture as the eluent. Follow the TLC detection (developing solvent is a 1:1 volume ratio of petroleum ether and ethyl acetate mixture). Collect the eluent containing the compound shown in formula (IB-5) according to the TLC detection. Evaporate the solvent from the collected eluent and dry to obtain a white solid product, namely compound (IB-5), with a yield of 48% (based on the molar amount of 2,4-dichloroquinazoline (II)) and a melting point of 269-271℃. 1 HNMR(500MHz,DMSO-d6)δ10.19(s,1H),10.03(s,1H),8.55(d,J=8.0Hz,1H),8.10(dd,J=8.0,1.3Hz,1H),7.91-7.87(m,2H),7.72(s,1H) ,7.67-7.63(m,2H),7.59-7.56(m,1H),7.25-7.21(m,1H),6.99(t,J=7.7Hz,1H),6.52-6.31(m,3H),3.87(d,J=5.9Hz,2H),2.20(s,3H).

[0086] Example 23: In vitro test of anticancer activity

[0087] Compounds (IA-1) to (IA-9) were subjected to bioactivity tests on human breast cancer cell line MDA-MB-231, human liver cancer cell line Huh7, and human liver cancer cell line HepG2, respectively. Compound (IA-1) was prepared by the method in Example 3. Compounds (IB-1) to (IB-5) were subjected to bioactivity tests on human breast cancer cell line MDA-MB-231, human liver cancer cell line Huh7, and human liver cancer cell line HepG2, respectively. Compound (IB-1) was prepared by the method in Example 15.

[0088] Test method: Tetraazole salt reduction method (MTT method).

[0089] Cell lines: human breast cancer cell line MDA-MB-231, human liver cancer cell line Huh7, and human liver cancer cell line HepG2. These cell lines were purchased from the Cell Bank of the Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

[0090] The experimental steps are as follows:

[0091] (1) Culture of tumor cells

[0092] All cell passages used in the experiments were no more than 5 generations. All consumables and reagents used in cell culture underwent strict sterilization procedures, and all experimental operations were performed in a sterile operating room.

[0093] (a) Recovery of tumor cells

[0094] Before starting the experiment, place all necessary items, such as culture flasks, centrifuge tubes, pipettes, pipette tips, and waste container, into the laminar flow hood and turn on the UV lamp for sterilization. Preheat the reagents used for cell culture in a 37°C water bath. After all preparations are complete, turn off the UV lamp and turn on the laminar flow hood's fluorescent light and ventilation fan. Remove the frozen tumor cells from the -80°C freezer and thaw them rapidly in a 37°C water bath, ensuring that a small amount of cryopreservation solution remains undried. Spray with alcohol and place in the laminar flow hood. Immediately transfer all cells from the cryopreservation tubes to 15mL centrifuge tubes containing 1640 culture medium, and gently mix with a pipette. Centrifuge the tubes at 1000rpm for 5 minutes, and aspirate the supernatant in the laminar flow hood. Pipette 2mL of 1640 culture medium into a centrifuge tube containing cell pellet to prepare a cell suspension. Mix the cell suspension thoroughly and transfer it to a flask with a bottom area of ​​25cm². 2 Add 4 mL of 1640 culture medium to the breathable culture flask, gently shake the culture flask to mix the cells, and place the culture flask in a 5% CO2, 37℃ constant temperature incubator for culture.

[0095] (b) Passage of tumor cells

[0096] When the cells are in good growth condition and cover 70%–80% of the bottom of the culture flask, they can be passaged. All passage procedures are performed in a clean bench. After preparation, passage can begin. First, pipette out the existing culture medium from the flask into a waste container. Add 2 mL of PBS and wash repeatedly, then aspirate the PBS. Next, add approximately 600–700 μL of trypsin (containing 0.02% EDTA, phenol red, and 0.25% trypsin), ensuring the bottom of the flask is completely covered with trypsin. Gently shake the flask and incubate at 37°C for 1–2 minutes. Observe cell detachment under a microscope. If a small number of cells remain attached to the flask wall, gently tap the flask wall with your fingertip until most cells detach from the bottom. Add 2 mL of 1640 culture medium to stop digestion, then gently pipette to remove all cells from the bottom of the flask. Transfer the cell suspension to a 15 mL centrifuge tube and centrifuge at 1000 rpm for 5 minutes. Remove the supernatant, dilute with 1640 culture medium, mix thoroughly by pipetting, and then evenly distribute into 2-3 culture flasks. Continue to incubate in a 5% CO2, 37℃ constant temperature incubator.

[0097] (c) Cryopreservation of tumor cells

[0098] Prepare the cell cryopreservation solution in advance according to the volume ratio of FBS (fetal bovine serum):DMSO = 9:1 and store it in a 4°C freezer for later use. All cryopreservation procedures are performed in a clean bench. After preparation, cryopreservation can begin. First, aspirate the original culture medium from the culture flask, add 2 mL of PBS buffer, wash repeatedly several times and discard. Then add approximately 600–700 μL of trypsin (containing 0.02% EDTA, phenol red, and 0.25% trypsin), gently shake the culture flask to ensure the trypsin covers the bottom, and incubate at 37°C for 1–2 minutes. Observe cell detachment under a microscope. If a small number of cells remain attached to the flask wall, gently tap the flask wall with your fingertip until most cells detach from the bottom. Add 2 mL of 1640 culture medium to stop digestion, then gently pipette the cells to remove them from the bottom of the flask. Transfer the cell suspension to a 15 mL centrifuge tube and centrifuge at 1000 rpm for 5 minutes. Discard the supernatant. Add 1 mL of cryopreservation solution just taken from the 4°C freezer, and pipette to form a cell suspension. Transfer the suspension to cryovials. After labeling the cell type, cell passage number, and cryopreservation date, incubate at 4°C for 30 min, then at -20°C for 1 hour, and finally store in an ultra-low temperature freezer at -80°C.

[0099] (2) MTT experimental method

[0100] (a) Cell counting: After digestion and centrifugation, the tumor cells in good growth condition in the culture flask were resuspended in 4 mL of 1640 culture medium. 10 μL of the cell suspension was transferred to a cell counting chamber for counting. After counting, B16F10 cells were diluted to 5 × 10⁻⁶. 4 per mL.

[0101] (b) Plating: Take a 96-well plate, add 100 μL of diluted cell suspension to the experimental wells, add 100 μL of 1640 culture medium to the blank wells, add 100 μL of PBS buffer around the plate, and incubate in a CO2 incubator.

[0102] (c) Compound preparation and drug administration: Dilute the 10 μmol / mL compound prepared in DMSO and the positive control drug sorafenib to the specified concentrations of 40 μM, 20 μM, 10 μM, 5 μM, 2.5 μM, and 1.25 μM using 1640 culture medium. Add the drugs using the medium exchange method. Discard the original culture medium and add 100 μL of 1640 culture medium containing the compound or positive control drug to each well. Add 100 μL of 1640 culture medium to each control and blank well. After drug administration, place the 96-well plate in a 5% CO2, 37°C incubator for further incubation.

[0103] (d) Add MTT: After 48 hours, take out the 96-well plate and place it in a clean bench. Add 10 μL of 5 mg / mL MTT solution to each well under light-protected conditions, and then place it in a 5% CO2, 37℃ constant temperature incubator for continued incubation.

[0104] (e) Detection: After incubating the 96-well plate with MTT in an incubator for 3.5-4 hours, carefully aspirate the solution from each well, add 150 μL of DMSO to each well to dissolve the generated formazan, and then place it on a plate shaker and shake for 20 minutes. Use a microplate reader to detect the absorbance value at 490 nm.

[0105] (f) Experimental data processing: Cell viability was calculated using the following formula. The value of 50% cell viability was the IC50 value. 50 .

[0106] Cell viability (%) = [(As-Ab) / (Ac-Ab)] × 100%

[0107] As: Experimental wells (containing cell culture medium, MTT, and toxic substances (i.e., compounds (IA) or (IB)))

[0108] Ac: Control wells (containing cell culture medium, MTT, and no toxic substances)

[0109] Ab: Blank wells (culture medium containing MTT but free of cells and toxic substances).

[0110] The test results are shown in Table 1:

[0111] Table 1. Inhibitory effects of compound (IA) on cancer cell growth (IC50, 110, 120, 130, 140, 150, 160, 170, 180, 19 ... 50 (μM)

[0112] compound MDA-MB-231 HepG2 Huh7 IA-1 ≥40 9.10±0.28 ≥40 IA-2 ≥40 ≥40 ≥40 IA-3 ≥40 14.93±1.73 ≥40 IA-4 10.69±0.84 ≥40 ≥40 IA-5 7.71±0.48 ≥40 ≥40 IA-6 ≥40 9.08±0.82 13.59±2.34 IA-7 ≥40 ≥40 ≥40 IA-8 ≥40 12.35±1.04 ≥40 IA-9 ≥40 ≥40 ≥40 Sorafenib 9.14±0.82 8.15±0.26 -

[0113] Table 2. Inhibitory effects of compound (ⅠB) on cancer cell growth (IC) 50 (μM)

[0114]

[0115]

Claims

1. A phenylurea-quinazoline compound, characterized in that: The structures of the phenylurea-quinazoline compounds are shown in formula (IA) or (IB): In formula (IA), R is H, 4-methoxy, 2,5-difluoro, or 3-methyl; in formula (IB), R is H, 3-trifluoromethyl-4-chloro, 4-chloro, or 3-methoxy.

2. A method for preparing a phenylurea-quinazoline compound as described in claim 1, characterized in that: The preparation method includes: (1) adding the compound shown in formula (II) to organic solvent A, stirring to dissolve, and adding dropwise a mixture containing the compound shown in formula (IIIA) or (IIIB), catalyst B and organic solvent A under stirring conditions at room temperature. After the addition is complete, the reaction solution is heated to reflux and stirred for 0.5 to 12 hours. The reaction solution is then separated and purified to obtain the phenylureaquinazoline compound shown in formula (IA) or (IB). The organic solvent A is one of the following: dichloromethane, ethanol, isopropanol, or toluene; Catalyst B is one of the following: triethylamine, 4-dimethylaminopyridine, pyridine, or sodium hydroxide; In equation (IIIA), the definition of R is the same as in equation (IA); in equation (IIIB), the definition of R is the same as in equation (IB).

3. The preparation method according to claim 2, characterized in that: The molar ratio of the compound shown in formula (II), formula (IIIA) or (IIIB) to the catalyst B is 1:0.8 to 1.2:0.1 to 1.

4. The use of the phenylureaquinazoline compound as described in claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a drug for the prevention or treatment of liver cancer.

5. The application as described in claim 4, characterized in that: The aforementioned drug for the prevention or treatment of liver cancer is a drug for the prevention or treatment of HepG2 human liver cancer cells.

6. The application as described in claim 4, characterized in that: The drug for preventing or treating liver cancer is a drug for preventing or treating human liver cancer cells Huh7; in the structural formula (IA) of the phenylureaquinazoline compound, R is 2,5-difluoro; in formula (IB), R is 3-trifluoromethyl-4-chloro or 3-methoxy.

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