CN117800925A - Phenyl ureido quinazoline compound, preparation method thereof and application thereof in preparation of medicines for preventing or treating liver cancer - Google Patents

Phenyl ureido quinazoline compound, preparation method thereof and application thereof in preparation of medicines for preventing or treating liver cancer Download PDF

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CN117800925A
CN117800925A CN202311388524.8A CN202311388524A CN117800925A CN 117800925 A CN117800925 A CN 117800925A CN 202311388524 A CN202311388524 A CN 202311388524A CN 117800925 A CN117800925 A CN 117800925A
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郑翔
赵江婷
饶国武
靳浩
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Zhejiang University of Technology ZJUT
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Zhejiang University of Technology ZJUT
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Abstract

The invention discloses phenyl ureido quinazoline compounds, preparation and application thereof in preparing medicaments for preventing or treating liver cancer. The structure of the phenylureido quinazoline compound is shown as a formula (IA) or (IB), wherein in the formula (IA), R is H, 4-methoxy, 2, 5-difluoro or 3-methyl; in the formula (IB), R is H, 3-trifluoromethyl-4-chloro, or 3-methoxy. The phenylureido quinazoline compound provided by the invention has a good inhibition effect on human liver cancer cells; the preparation method of the compound is simple and convenient, is easy to operate, has easily available raw materials and lower production cost, and is suitable for industrial application.

Description

Phenyl ureido quinazoline compound, preparation method thereof and application thereof in preparation of medicines for preventing or treating liver cancer
Technical Field
The invention relates to a quinazoline compound and application thereof, in particular to a phenyl ureido quinazoline compound and a preparation method thereof, and application of the compound or pharmaceutically acceptable salt thereof in preparing medicaments for preventing or treating liver cancer.
Background
The quinazoline compounds have a plurality of better biological activities, have wide application in the field of medicine, and particularly have obvious antiviral activity, antibacterial activity, antitumor activity and the like, and have been marketed as antitumor drugs. Such as Gefitinib (Gefitinib) and Erlotinib (Erlotinib), which are marketed for the treatment of lung cancer, and Lapatinib (Lapatinib) for the treatment of breast cancer, all of which belong to the quinazoline class. Novel quinazoline compounds and their biological activity are also reported in the literature (see Y.—Y. Ke, H.— Y.Shiao, Y.C.Hsu, C.—Y. Chu, W.—C. Wang, Y.—C.Lee, W.—H. Lin, C.— H.Chen, J.T.A.Hsu, C.—W.Chang, C.—W.Lin, T.—K.Yeh, Y.— S.Chao, M.S.Coumar, 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, 9-1204). Of course, most quinazoline compounds do not possess antitumor activity.
Disclosure of Invention
The invention aims to provide a novel quinazoline compound-phenyl ureido quinazoline compound with anticancer activity, a preparation method and application thereof, and the compound has good inhibition effect on human liver cancer cells at a certain dosage; the preparation method of the compound is simple and convenient, is easy to operate, has easily available raw materials and lower production cost, and is suitable for industrial application.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
in a first aspect, the invention provides a phenylureido quinazoline compound represented by formula (IA) or (IB),
in the formula (IA), R is H, 4-methoxy, 2, 5-difluoro or 3-methyl, and the specific structure is as follows:
in the formula (IB), R is H, 3-trifluoromethyl-4-chlorine, 4-chlorine or 3-methoxy, and the specific structure is as follows:
in a second aspect, the invention provides a method for preparing a phenylureido quinazoline compound, which comprises the following steps: (1) Adding a compound shown in a formula (II) into an organic solvent A, stirring and dissolving, dropwise adding a mixed solution containing the compound shown in a formula (IIIA) or (IIIB), a catalyst B and the organic solvent A under the condition of stirring at room temperature, heating the reaction solution to a reflux state after the dripping is finished, stirring and reacting for 0.5-12 hours, and separating and purifying the reaction solution to obtain phenylureido quinazoline compounds shown in a formula (IA) or (IB);
the organic solvent A is one of the following: dichloromethane, ethanol, isopropanol or toluene;
the catalyst B is one of the following: triethylamine, 4-Dimethylaminopyridine (DMAP), pyridine or sodium hydroxide;
in the formula (IIIA), R is defined as in the formula (IA); in formula (IIIB), R is as defined in formula (IB).
Further, the ratio of the amount of the compound represented by the formula (II), the compound represented by the formula (IIIA) or (IIIB) to the amount of the catalyst B to be fed is 1:0.8 to 1.2:0.1 to 1.
Further, the amount of the organic solvent A to be used may be 10 to 30mL/mmol in terms of the amount of the substance capable of dissolving the compound represented by the formula (II), the compound represented by the formula (IIIA) or (IIIB), and the catalyst B, and preferably the amount of the organic solvent A to be used for dissolving the compound represented by the formula (II) may be 4 to 10mL/mmol in terms of the amount of the substance capable of dissolving the compound represented by the formula (II), and the amount of the organic solvent A to be used for dissolving the compound represented by the formula (IIIA) or (IIIB) and the catalyst B may be 6 to 20mL/mmol in terms of the amount of the substance capable of dissolving the compound represented by the formula (II).
Further, the reaction process is followed by TLC (developing agent is petroleum ether and ethyl acetate mixed solution with volume ratio of 0.5-10:1) to determine the reaction end point, and the reaction time is generally 0.5-12 hours.
Further, the separation and purification in the step (1) of the invention adopts the following steps: after the reaction is finished, evaporating the solvent, and performing column chromatography on the residue to obtain the phenylureido quinazoline compound.
Further, the operation steps of the column chromatography in the step (1) of the present invention are specifically as follows: taking residues after solvent evaporation in a single-mouth bottle, adding an organic solvent C to dissolve the residues to obtain a dissolution solution, adding column chromatography silica gel (preferably 300-400 mesh coarse pore (zcx.II) column chromatography silica gel) with the mass of 1-4 times of the residues into the dissolution solution, uniformly mixing, evaporating the solvent to obtain a mixture of the dried residues and the silica gel, loading the mixture into a column, and loading the sample into a sample with the volume ratio of 0.5-10: 1, eluting with petroleum ether and ethyl acetate mixed solution as eluent, performing TLC tracking detection (developing agent is petroleum ether and ethyl acetate mixed solution with volume ratio of 0.5-10:1), collecting eluent containing phenylureido quinazoline compounds according to TLC detection, concentrating and drying the eluent to obtain phenylureido quinazoline compounds; the organic solvent C is one of the following: petroleum ether, dichloromethane, chloroform or ethyl acetate; the organic solvent C is used in an amount to dissolve the residue.
The organic solvents A and C are organic solvents, so that the organic solvents used in different steps are conveniently distinguished and named, and letters have no meaning.
In a third aspect, the invention also relates to application of the phenylureido quinazoline compound or the pharmaceutically acceptable salt thereof in preparing medicaments for preventing or treating liver cancer.
Preferably, the medicine for preventing or treating liver cancer is a medicine for preventing or treating human liver cancer cell HepG2.
Preferably, the medicine for preventing or treating liver cancer is a medicine for preventing or treating human liver cancer cell Huh 7; in the structural formula (IA) of the phenylureido quinazoline compound, R is 2, 5-difluoro (namely a compound (IA-6)); in formula (IB), R is 3-trifluoromethyl-4-chloro or 3-methoxy (i.e., compound (IB-2) or (IB-4)).
In particular, the compounds (IB-2) and (IB-3) provided by the invention also have good inhibition effect on breast cancer.
The term "pharmaceutically acceptable" in this application means: the compounds are chemically and/or toxicologically compatible with the other ingredients comprising the formulation and/or with the human or mammal with which the disease or condition is to be prevented or treated.
The term "pharmaceutically acceptable salt" refers to the relatively non-toxic, inorganic or organic acid addition salts of the compounds of the present invention. See, for example, S.M. Bere et al, "Pharmaceutical Salts", J.Pharm. Sci.1977, 66,1-19. Among them, inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid or nitric acid, etc.; organic acids such as formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, caproic 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, pamoic acid, pectate acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfuric 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, glucoheptonic acid, glycerophosphate, aspartic acid, sulfosalicylic acid, and the like.
The beneficial effects of the invention are mainly as follows: (1) The phenylureido quinazoline compound (I) has good anti-liver cancer activity and is expected to be applied to preparation of medicines for preventing or treating liver cancer; (2) The preparation method of the phenyl ureido quinazoline compound provided by the invention is simple and easy to operate, raw materials are easy to obtain, the production cost is low, and the preparation method is suitable for practicality.
Detailed Description
The invention is further illustrated by reference to specific examples, which are given below to illustrate the invention and are not to be construed as limiting the invention in any way.
The specific conditions are not noted in the examples of the present invention, and are carried out according to conventional conditions or conditions suggested by the manufacturer. The reagents or apparatus used are conventional products, which are available by conventional technical means or commercially available, without the manufacturer's knowledge.
Preparation of Compound (II) reference (Journal ofMedicinal Chemistry,2022,65 (10), 7246-7261;Bioorganic Chemistry,2021,117,105407).
The specific structure of the compound (IIIA) is as follows:
the specific structure of the compound (IIIB) is as follows:
the specific structure of compound (IB) is as follows:
example 1: preparation of Compound (IIIA-1)
1.12g (12.0 mmol) of aniline and 1.22g (10.0 mmol) of DMAP are added to 30mL of methylene chloride and stirred, 30mL of methylene chloride solution containing 1.64g (10.0 mmol) of 4-nitrophenyl isocyanate is added dropwise at 0-5 ℃, the mixture is reacted at room temperature for 20h, water is washed (25 mL. Times.3), the organic phase is separated, dried, filtered and the solvent is distilled off in vacuo, and the residue is recrystallized from ethanol to obtain 1- (4-nitrophenyl) -3-phenylurea in 71% yield.
1.29g (5.0 mmol) of 1- (4-nitrophenyl) -3-phenylurea was added to 30mL of ethanol, 0.20g of Pd/C (5%) was added thereto, and under stirring at room temperature, hydrogen was introduced under normal pressure to react, TLC detection (developing agent: petroleum ether and ethyl acetate mixed solution in a volume ratio of 1:1) was carried out until the reaction was completed, filtration was carried out, the solvent was distilled off in vacuo, and the residue was recrystallized from ethanol to give the product (IIIA-1) in 83% yield.
Preparation of Compounds (IIIA-2) to (IIIA-9): the aniline was replaced with the corresponding substituted aniline, and the corresponding compounds (IIIA-2) to (IIIA-9) were prepared by the method described in example 1, and are not described in detail herein.
Example 2: preparation of Compound (IIIB-1)
1.29g (12.0 mmol) of benzylamine and 1.22g (10.0 mmol) of DMAP were added to 30mL of methylene chloride and stirred, 30mL of a methylene chloride solution containing 1.64g (10.0 mmol) of 4-nitrophenyl isocyanate was added dropwise at 0 to 5℃and reacted at room temperature for 22 hours, water was washed (25 mL. Times.3), the organic phase was separated, dried, filtered and the solvent was distilled off in vacuo, and the residue was recrystallized from ethanol to give 1- (4-nitrophenyl) -3-benzylurea in 65% yield.
1.36g (5.0 mmol) of 1- (4-nitrophenyl) -3-benzyl urea was added to 30mL of ethanol, 0.20g of Pd/C (5%) was added thereto, and under stirring at room temperature, hydrogen was introduced under normal pressure to react, TLC detection (developing agent: petroleum ether and ethyl acetate mixed solution in a volume ratio of 1:1) was carried out until the reaction was completed, filtration was carried out, the solvent was distilled off in vacuo, and the residue was recrystallized from ethanol to give the product (IIIB-1) in 78% yield.
Preparation of Compounds (IIIB-2) to (IIIB-5): the corresponding compounds (IIIB-2) to (IIIB-5) were prepared by substituting benzylamine with the corresponding substituted benzylamine by the method of example 2, and are not described in detail herein.
Example 3: preparation of Compound (IA-1)
20mL of methylene chloride is dissolved into 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), 30mL of methylene chloride solution containing 0.455g (2.0 mmol) of compound (IIIA-1) and 0.122g (1.0 mmol) of DMAP is dropwise added under the condition of magnetic stirring at room temperature, the reaction liquid is heated to reflux in an oil bath after the completion of the dropwise addition, the reflux reaction is carried out for 12 hours (the reaction process is tracked and detected by TLC, and the developing agent is petroleum ether and ethyl acetate in volume ratio of 1:2)Mixing the solution), evaporating the solvent from the reaction solution, performing column chromatography on the residue, namely adding 10 milliliters of petroleum ether solvent into the residue after evaporating the solvent to dissolve the residue to obtain a solution, adding 1.5 grams of silica gel (300-400 meshes of coarse pore (zcx.II) column chromatography silica gel) into the solution, uniformly mixing, evaporating the solvent to obtain a mixture of the dried residue and the silica gel, filling the mixture into a column, and then carrying out the column mixing according to the volume ratio of 1:2 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:2), collecting eluent containing the compound shown in formula (IA-1) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IA-1), with yield of 63% (based on the amount of 2, 4-dichloroquinazoline (II)) and melting point of 218-220 ℃. 1 HNMR(500MHz,DMSO-d 6 )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＝7.9Hz,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-d 6 )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.
Example 4: preparation of Compound (IA-1)
8mL of toluene is dissolved into 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), 12mL of toluene solution containing 0.545g (2.4 mmol) of compound (IIIA-1) and 0.008g (0.2 mmol) of sodium hydroxide is dropwise added under the condition of magnetic stirring at room temperature, the reaction solution is heated to reflux in an oil bath after the completion of the dripping, the reflux reaction is carried out for 0.5 hour (the reaction process adopts TLC tracking detection, the developing agent is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 10:1), the solvent is distilled off from the reaction solution, the residue is subjected to column chromatography, namely, the residue after the solvent is distilled off is taken, 10mL of dichloromethane solvent is added to dissolve the residue to obtain a dissolving solution, 1.5 g of silica gel (300-400 meshes of coarse pore (zcx.II) column chromatography silica gel) is added into the dissolving solution, after the solvent is distilled off, a mixture of the dried residue and the silica gel is obtained, the mixture is packed into a column, and the volume ratio of 10:1 in petroleum ether and ethyl acetate as eluent, eluting, and performing TLC tracking detection (developing agent is volume ratio)10:1 with ethyl acetate), collecting the eluent containing the compound shown in the formula (IA-1) according to TLC detection, evaporating the solvent from the collected eluent, and drying to obtain a white solid product, namely the compound (IA-1), wherein the yield is 45% (based on the amount of 2, 4-dichloro quinazoline (II) and the melting point is 218-220 ℃. 1 H NMR 13 C NMR was as in example 1.
Example 5: preparation of Compound (IA-1)
20mL of ethanol is dissolved into 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), 40mL of ethanol solution containing 0.364g (1.6 mmol) of compound (IIIA-1) and 0.202g (2.0 mmol) of triethylamine is dropwise added under the condition of magnetic stirring at room temperature, the reaction liquid is heated to reflux in an oil bath after the completion of the dripping, the reflux reaction is carried out for 6 hours (the reaction process adopts TLC tracking detection, the developing agent is a petroleum ether and ethyl acetate mixed solution with the volume ratio of 1:1), the solvent is distilled off from the reaction liquid, the residue is subjected to column chromatography, namely, the residue after the solvent is distilled off is taken, 10mL of chloroform solvent is added to dissolve the residue to obtain a solution, then 1.5 g of silica gel (300-400 meshes of coarse pore (zcx.II) column chromatography silica gel) is added into the solution, the solution is uniformly mixed, the solvent is distilled off to obtain a mixture of dry residue and the mixture is packed into a column, and then the volume ratio of 1:1 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:1), collecting eluent containing the compound shown in formula (IA-1) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IA-1), wherein the yield is 55% (based on the amount of compound (IIIA-1)) and the melting point is 218-220 ℃. 1 H NMR 13 C NMR was as in example 1.
Example 6: preparation of Compound (IA-1)
10mL of isopropanol was dissolved in 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), a solution of 0.455g (2.0 mmol) of compound (IIIA-1) and 0.079g (1.0 mmol) of pyridine in 15mL of isopropanol was added dropwise under magnetic stirring at room temperature, the reaction solution was heated to reflux in an oil bath after the completion of the dropping, the reflux reaction was carried out for 4 hours (the reaction process was detected by TLC tracking, the developing agent was a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 5:1), and the reaction was carried outEvaporating solvent, and performing residue column chromatography, namely adding 10 milliliters of ethyl acetate solvent into the residue after evaporating the solvent to dissolve the residue to obtain a dissolving solution, adding 1.5 grams of silica gel (300-400 meshes of coarse pore (zcx.II) column chromatography silica gel) into the dissolving solution, uniformly mixing, evaporating the solvent to obtain a mixture of the dried residue and the silica gel, filling the mixture into a column, and then carrying out volume ratio (5): 1 as eluent, TLC tracking detection (developing solvent is mixed solution of petroleum ether and ethyl acetate in volume ratio of 5:1), collecting eluent containing the compound shown in formula (IA-1) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IA-1), wherein the yield is 42% (based on the amount of 2, 4-dichloroquinazoline (II)) and the melting point is 218-220 ℃. 1 H NMR 13 C NMR was as in example 1.
Example 7: preparation of Compound (IA-2)
20mL of methylene chloride is dissolved into 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), 30mL of methylene chloride solution containing 0.511g (2.0 mmol) of compound (IIIA-2) and 0.122g (1.0 mmol) of DMAP is dropwise added under the condition of magnetic stirring at room temperature, the reaction liquid is heated to reflux in an oil bath after the completion of the dropwise addition, the reflux reaction is carried out for 10 hours (the reaction process adopts TLC tracking detection, the developing agent is a petroleum ether and ethyl acetate mixed solution with the volume ratio of 1:1), the solvent is distilled off from the reaction liquid, the residue is subjected to column chromatography, namely, the residue after the solvent is distilled off is taken, 10mL of petroleum ether solvent is added to dissolve the residue to obtain a dissolving liquid, then 1.5 g of silica gel (300-400-mesh coarse pore (zcx.II) column chromatography silica gel) is added into the dissolving liquid, after the solvent is distilled off, a mixture of the dried residue and the silica gel is obtained, the mixture is packed into a column with the volume ratio of 1:1 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:1), collecting eluent containing the compound shown in formula (IA-2) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IA-2), with yield of 61% (based on the amount of 2, 4-dichloroquinazoline (II)) and melting point of 232-234 ℃. 1 HNMR(500MHz,DMSO-d 6 )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).
Example 8: preparation of Compound (IA-3)
20mL of methylene chloride is dissolved into 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), 30mL of methylene chloride solution containing 0.515g (2.0 mmol) of compound (IIIA-3) and 0.122g (1.0 mmol) of DMAP is dropwise added under the condition of magnetic stirring at room temperature, the reaction liquid is heated to reflux in an oil bath after the completion of the dropwise addition, the reflux reaction is carried out for 9 hours (the reaction process adopts TLC tracking detection, the developing agent is a petroleum ether and ethyl acetate mixed solution with the volume ratio of 1:1), the solvent is distilled off from the reaction liquid, the residue is subjected to column chromatography, namely, the residue after the solvent is distilled off is taken, 10mL of petroleum ether solvent is added to dissolve the residue to obtain a dissolving liquid, then 1.5 g of silica gel (300-400 meshes of coarse pore (zcx.II) column chromatography silica gel) is added into the dissolving liquid, after the solvent is distilled off, the mixture of the dried residue and the silica gel is obtained, the mixture is packed into a column with the volume ratio of 1:1 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:1), collecting eluent containing the compound shown in formula (IA-3) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IA-3), with yield of 68% (based on the amount of 2, 4-dichloroquinazoline (II)) and melting point of 265-267 ℃. 1 HNMR(500MHz,DMSO-d 6 )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-d 6 )δ163.0,159.6,156.6,154.6,153.0,150.8,150.5,141.0,137.3,133.0,132.0,124.1,122.5,120.1,118.3,114.2,113.9,55.4.
Example 9: preparation of Compound (IA-4)
20mL of methylene chloride was dissolved in 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), and under magnetic stirring at room temperature, a solution containing 0.592g (2.0 mmol) of the compound (IIIA-4) and 0.122 was added dropwise30mL of dichloromethane solution of g (1.0 mmol) DMAP is heated to reflux in an oil bath after dripping, reflux reaction is carried out for 10 hours (TLC tracking detection is adopted in the reaction process, the developing agent is petroleum ether and ethyl acetate mixed solution with the volume ratio of 1:1), the solvent is distilled off from the reaction solution, the residue is subjected to column chromatography, namely 7 mL of petroleum ether solvent is added into the residue after the solvent is distilled off to dissolve the residue, a dissolving solution is obtained, then 1.5 g of silica gel (300-400 meshes of coarse pore (zcx.II) type column chromatography silica gel) is added into the dissolving solution, after uniform mixing, the solvent is distilled off to obtain a mixture of the dried residue and the silica gel, the mixture is packed into a column, and then the volume ratio of the mixture is 1:1 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:1), collecting eluent containing the compound shown in formula (IA-4) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IA-4), with yield of 51% (based on the amount of 2, 4-dichloroquinazoline (II)) and melting point of 188-189 ℃. 1 HNMR(500MHz,DMSO-d 6 )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-d 6 )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.
Example 10: preparation of Compound (IA-5)
Dissolving 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II) in 20mL of dichloromethane, dropwise adding 30mL of dichloromethane solution containing 0.511g (2.0 mmol) of compound (IIIA-5) and 0.122g (1.0 mmol) of DMAP under the condition of magnetic stirring at room temperature, heating the reaction liquid to reflux after the completion of the dripping, carrying out reflux reaction for 8 hours (TLC tracking detection is adopted in the reaction process, the developing agent is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 1:1), evaporating the reaction liquid to remove the solvent, carrying out column chromatography on the residue, namely, taking the residue after evaporating the solvent, adding 10mL of petroleum ether solvent to dissolve the residue to obtain a solution, then adding 1.5 g of silica gel (300-400 meshes of coarse pore (zcx.II) column chromatography silica gel) into the solution, uniformly mixing, and evaporating the solvent to obtain a mixture of the dried residue and the silica gelThe mixture was packed in a column and then in a volume ratio of 1:1 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:1), collecting eluent containing the compound shown in formula (IA-5) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IA-5), with yield of 67% (based on the amount of 2, 4-dichloroquinazoline (II)) and melting point of 204-206 ℃. 1 HNMR(500MHz,DMSO-d 6 )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-d 6 )δ159.6,156.6,152.7,150.9,139.8,137.9,137.0,134.1,132.2,127.1,127.0,126.7,124.1,123.6,118.4,116.1,113.9,21.3.
Example 11: preparation of Compound (IA-6)
20mL of methylene chloride is dissolved into 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), 30mL of methylene chloride solution containing 0.526g (2.0 mmol) of compound (IIIA-6) and 0.122g (1.0 mmol) of DMAP is dropwise added under the condition of magnetic stirring at room temperature, the reaction liquid is heated to reflux in an oil bath after the completion of the dropwise addition, the reflux reaction is carried out for 11 hours (the reaction process adopts TLC tracking detection, the developing agent is a petroleum ether and ethyl acetate mixed solution with the volume ratio of 1:1), the solvent is distilled off from the reaction liquid, the residue is subjected to column chromatography, namely, the residue after the solvent is distilled off is taken, 10mL of petroleum ether solvent is added to dissolve the residue to obtain a dissolving liquid, then 1.5 g of silica gel (300-400 meshes of coarse pore (zcx.II) column chromatography silica gel) is added into the dissolving liquid, after the solvent is distilled off, the mixture of the dried residue and the silica gel is obtained, the mixture is packed into a column with the volume ratio of 1:1 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:1), collecting eluent containing the compound shown in formula (IA-6) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IA-6), with yield of 67% (based on the amount of 2, 4-dichloroquinazoline (II)) and melting point of 238-239 ℃. 1 HNMR(500MHz,DMSO-d 6 )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-d 6 )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.
Example 12: preparation of Compound (IA-7)
20mL of methylene chloride is dissolved into 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), 30mL of methylene chloride solution containing 0.523g (2.0 mmol) of compound (IIIA-7) and 0.122g (1.0 mmol) of DMAP is dropwise added under the condition of magnetic stirring at room temperature, the reaction liquid is heated to reflux in an oil bath after the completion of the dropwise addition, the reflux reaction is carried out for 6 hours (the reaction process adopts TLC tracking detection, the developing agent is a petroleum ether and ethyl acetate mixed solution with the volume ratio of 1:1), the solvent is distilled off from the reaction liquid, the residue is subjected to column chromatography, namely, the residue after the solvent is distilled off is taken, 10mL of petroleum ether solvent is added to dissolve the residue to obtain a dissolving liquid, then 1.5 g of silica gel (300-400-mesh coarse pore (zcx.II) column chromatography silica gel) is added into the dissolving liquid, after the solvent is distilled off, a mixture of the dried residue and the silica gel is obtained, the mixture is packed into a column with the volume ratio of 1:1 as eluent, TLC tracking detection (developing solvent is mixed solution of petroleum ether and ethyl acetate in volume ratio of 1:1), collecting eluent containing a compound shown in formula (IA-7) according to TLC detection, evaporating the solvent from the collected eluent, and drying to obtain a white solid product, namely the compound (IA-7), wherein the yield is 59% (based on the amount of 2, 4-dichloroquinazoline (II)) and the melting point is 198-199 ℃. 1 HNMR(500MHz,DMSO-d 6 )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),7.85(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-d 6 )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.
Example 13: preparation of Compound (IA-8)
Will be 20Dissolving 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II) in 30mL of dichloromethane solution containing 0.483g (2.0 mmol) of compound (IIIA-8) and 0.122g (1.0 mmol) of DMAP under the condition of magnetic stirring at room temperature, heating the reaction liquid to reflux after the completion of the dripping, carrying out reflux reaction for 5 hours (TLC tracking detection is adopted in the reaction process, developing agent is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 1:1), evaporating the solvent from the reaction liquid, carrying out column chromatography on the residue, namely, taking the residue after evaporating the solvent, adding 10mL of petroleum ether solvent to dissolve the residue to obtain a solution, then adding 1.5 g of silica gel (300-400-mesh coarse pore (zcx.II) column chromatography silica gel) into the solution, uniformly mixing, evaporating the solvent to obtain a mixture of dried residue and the silica gel, filling the mixture into a column, and then carrying out the column according to a volume ratio of 1:1 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:1), collecting eluent containing the compound shown in formula (IA-8) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IA-8), wherein the yield is 55% (based on the amount of 2, 4-dichloroquinazoline (II)) and the melting point is 182-184 ℃. 1 HNMR(500MHz,DMSO-d 6 )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).
Example 14: preparation of Compound (IA-9)
Dissolving 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II) in 20mL of dichloromethane, dropwise adding 30mL of dichloromethane solution containing 0.515g (2.0 mmol) of compound (IIIA-9) and 0.122g (1.0 mmol) of DMAP under the condition of magnetic stirring at room temperature, heating the reaction liquid to reflux in an oil bath after the completion of the dripping, carrying out reflux reaction for 9 hours (TLC tracking detection is adopted in the reaction process, the developing agent is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 1:1), evaporating the reaction liquid to remove the solvent, carrying out column chromatography on the residue, namely, taking the residue after evaporating the solvent, adding 10mL of petroleum ether solvent to dissolve the residue to obtain a solution, then adding 1.5 g of silica gel (300-400 meshes of coarse pore (zcx.II) column chromatography silica gel) into the solution, uniformly mixing, and then evaporating the solventA mixture of dry residue and silica gel was obtained, the mixture was packed in a column, and then, in a volume ratio of 1:1 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:1), collecting eluent containing the compound shown in formula (IA-9) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IA-9), with yield of 51% (based on the amount of 2, 4-dichloroquinazoline (II)) and melting point of 228-230 ℃. 1 HNMR(500MHz,DMSO-d 6 )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).
Example 15: preparation of Compound (IB-1)
20mL of methylene chloride is dissolved into 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), 30mL of methylene chloride solution containing 0.483g (2.0 mmol) of compound (IIIB-1) and 0.122g (1.0 mmol) of DMAP is dropwise added under the condition of magnetic stirring at room temperature, the reaction liquid is heated to reflux in an oil bath after the completion of the dropwise addition, the reflux reaction is carried out for 12 hours (the reaction process adopts TLC tracking detection, the developing agent is a petroleum ether and ethyl acetate mixed solution with the volume ratio of 1:2), the solvent is distilled off from the reaction liquid, the residue is subjected to column chromatography, namely, the residue after the solvent is distilled off is taken, 10mL of petroleum ether solvent is added to dissolve the residue to obtain a solution, then 1.5 g of silica gel (300-400 meshes of coarse pore (zcx.II) column chromatography silica gel) is added into the solution, after the solvent is evenly mixed, the mixture of the dried residue and the silica gel is obtained, and the mixture is packed into a column with the volume ratio of 1:2 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:2), collecting eluent containing compound shown in formula (IB-1) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IB-1), with yield of 70% (based on the amount of 2, 4-dichloroquinazoline (II)) and melting point of 189-191 ℃. 1 HNMR(500MHz,DMSO-d 6 )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).
Example 16: preparation of Compound (IB-1)
8mL of toluene is dissolved into 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), under the condition of magnetic stirring at room temperature, 12mL of toluene solution containing 0.579g (2.4 mmol) of compound (IIIB-1) and 0.008g (0.2 mmol) of sodium hydroxide is dropwise added, the reaction solution is heated to reflux in an oil bath after the completion of the dropwise addition, the reflux reaction is carried out for 0.5 hour (the reaction process adopts TLC tracking detection, the developing agent is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 10:1), the solvent is distilled off from the reaction solution, the residue is subjected to column chromatography, namely, the residue after the solvent is distilled off is taken, 10mL of dichloromethane solvent is added to dissolve the residue to obtain a solution, then 1.5 g of silica gel (300-400-mesh coarse pore (zcx.II) column chromatography silica gel) is added into the solution, after the solvent is distilled off, the mixture of the dried residue and the silica gel is packed into a column, and then the volume ratio of 10:1 as eluent, TLC tracking detection (developing solvent is mixed solution of petroleum ether and ethyl acetate in volume ratio of 10:1), collecting eluent containing the compound shown in formula (IB-1) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely the compound (IB-1), with yield of 56% (based on the amount of 2, 4-dichloroquinazoline (II)) and melting point of 189-191 ℃. 1 HNMR is the same as in example 1.
Example 17: preparation of Compound (IB-1)
Dissolving 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II) in 20mL of ethanol, dropwise adding 40mL of ethanol solution containing 0.386g (1.6 mmol) of compound (IIIB-1) and 0.202g (2.0 mmol) of triethylamine under the condition of magnetic stirring at room temperature, heating the reaction liquid to reflux after the completion of the dripping, carrying out reflux reaction for 6 hours (TLC tracking detection is adopted in the reaction process, developing solvent is a petroleum ether and ethyl acetate mixed solution with the volume ratio of 1:1), evaporating the reaction liquid to remove the solvent, carrying out column chromatography on the residue, namely, taking the residue after evaporating the solvent, adding 10mL of chloroform solvent to dissolve the residue to obtain a solution, and then adding 1.5 g of silica gel (300-400 meshes of coarse pore (zcx.II) column chromatography silica gel) into the solution) After mixing evenly, evaporating the solvent to obtain a mixture of the dried residue and silica gel, filling the mixture into a column, and then, carrying out volume ratio of 1:1 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:1), collecting eluent containing the compound shown in formula (IB-1) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IB-1), with yield of 67% (based on the amount of compound (IIIB-1)) and melting point of 189-191 ℃. 1 H NMR was as in example 1.
Example 18: preparation of Compound (IB-1)
10mL of isopropanol (2.0 mmol) is dissolved into 0.398g of 2, 4-dichloroquinazoline (II), 15mL of isopropanol solution containing 0.483g (2.0 mmol) of compound (IIIB-1) and 0.079g (1.0 mmol) of pyridine is dropwise added under the condition of magnetic stirring at room temperature, the reaction solution is heated to reflux in an oil bath after the completion of the dripping, the reflux reaction is carried out for 4 hours (the reaction process is detected by TLC tracking, the developing agent is a petroleum ether and ethyl acetate mixed solution with the volume ratio of 5:1), the solvent is distilled off from the reaction solution, the residue is subjected to column chromatography, namely, the distilled residue is taken and dissolved by adding 10mL of ethyl acetate solvent into the solvent to obtain a solution, then 1.5 g of silica gel (300-400-mesh coarse pore (zcx.II) type column chromatography silica gel) is added into the solution, after the solvent is evenly mixed, the mixture of the dried residue and the silica gel is packed into a column, and then the mixture is loaded into a column with the volume ratio of 5:1 as eluent, TLC tracking detection (developing solvent is mixed solution of petroleum ether and ethyl acetate in volume ratio of 5:1), collecting eluent containing the compound shown in formula (IB-1) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IB-1), with yield of 54% (based on the amount of 2, 4-dichloroquinazoline (II)) and melting point of 189-191 ℃. 1 H NMR was as in example 1.
Example 19: preparation of Compound (IB-2)
20mL of methylene chloride was dissolved in 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), and 30mL containing 0.687g (2.0 mmol) of compound (IIIB-2) and 0.122g (1.0 mmol) of DMAP was added dropwise under magnetic stirring at room temperatureHeating a dichloromethane solution in an oil bath until the reaction solution is refluxed after dripping, carrying out reflux reaction for 11 hours (TLC tracking detection is adopted in the reaction process, the developing agent is petroleum ether and ethyl acetate mixed solution with the volume ratio of 1:1), evaporating the solvent from the reaction solution, carrying out column chromatography on the residues, namely, taking residues after evaporating the solvent, adding 10 milliliters of petroleum ether solvent to dissolve the residues to obtain a dissolving solution, then adding 1.5 grams of silica gel (300-400 meshes of coarse pore (zcx.II) column chromatography silica gel) into the dissolving solution, uniformly mixing, evaporating the solvent to obtain a mixture of the dried residues and the silica gel, loading the mixture into a column, and then carrying out column chromatography according to the volume ratio of 1:1 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:1), collecting eluent containing the compound shown in formula (IB-2) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IB-2), with yield of 55% (based on the amount of 2, 4-dichloroquinazoline (II)) and melting point of 267-269 ℃. 1 HNMR(500MHz,DMSO-d 6 )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-d 6 )δ168.4,159.6,156.5,151.0,147.9,135.8,134.2,133.7,132.1,127.0,126.7,123.8,123.6,119.6,116.5,116.0,113.9,111.3,46.8,35.9.
Example 20: preparation of Compound (IB-3)
Dissolving 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II) in 20mL of dichloromethane, dropwise adding 30mL of dichloromethane solution containing 0.551g (2.0 mmol) of compound (IIIB-3) and 0.122g (1.0 mmol) of DMAP under the condition of magnetic stirring at room temperature, heating the reaction liquid to reflux in an oil bath after the completion of the dripping, carrying out reflux reaction for 10 hours (TLC tracking detection is adopted in the reaction process, the developing agent is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 1:1), evaporating the reaction liquid to remove the solvent, carrying out column chromatography on the residue, namely, taking the residue after the solvent is evaporated, adding 10mL of petroleum ether solvent to dissolve the residue to obtain a dissolving liquid, then adding 1.5 g of silica gel (300-400-mesh coarse pore (zcx.II) column chromatography silica gel) into the dissolving liquid, uniformly mixing, and evaporating the solvent to obtain a dry residueMixture with silica gel, column the mixture, then in volume ratio 1:1 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:1), collecting eluent containing compound shown in formula (IB-3) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IB-3), with yield of 67% (based on the amount of 2, 4-dichloroquinazoline (II)) and melting point of 271-273 ℃. 1 HNMR(500MHz,DMSO-d 6 )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-d 6 )δ159.6,156.7,155.4,151.0,138.8,137.8,134.1,131.5,130.9,130.8,128.4,127.0,126.6,124.1,123.5,117.9,113.9,35.3.
Example 21: preparation of Compound (IB-4)
20mL of methylene chloride is dissolved into 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), 30mL of methylene chloride solution containing 0.542g (2.0 mmol) of compound (IIIB-4) and 0.122g (1.0 mmol) of DMAP is dropwise added under the condition of magnetic stirring at room temperature, the reaction liquid is heated to reflux in an oil bath after the completion of the dropwise addition, the reflux reaction is carried out for 8 hours (the reaction process adopts TLC tracking detection, the developing agent is a petroleum ether and ethyl acetate mixed solution with the volume ratio of 1:1), the solvent is distilled off from the reaction liquid, the residue is subjected to column chromatography, namely, 7 mL of petroleum ether solvent is added into the residue after the solvent is distilled off to dissolve the residue, so as to obtain a dissolving liquid, then 1.5 g of silica gel (300-400 meshes of coarse pore (zcx.II) column chromatography silica gel) is added into the dissolving liquid, after the solvent is distilled off, so as to obtain a mixture of dried residue and the silica gel, the mixture is packed into a column with the volume ratio of 1:1 as eluent, TLC tracking detection (developing solvent is petroleum ether and ethyl acetate mixed solution with volume ratio of 1:1), collecting eluent containing compound shown in formula (IB-4) according to TLC detection, evaporating solvent from the collected eluent, and drying to obtain white solid product, namely compound (IB-4), with yield of 62% (based on the amount of 2, 4-dichloroquinazoline (II)) and melting point of more than 300 ℃. 1 H NMR(500MHz,DMSO-d 6 )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-d 6 )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.
Example 22: preparation of Compound (IB-5)
20mL of methylene chloride is dissolved into 0.398g (2.0 mmol) of 2, 4-dichloroquinazoline (II), 30mL of methylene chloride solution containing 0.511g (2.0 mmol) of compound (IIIB-5) and 0.122g (1.0 mmol) of DMAP is dropwise added under the condition of magnetic stirring at room temperature, the reaction liquid is heated to reflux in an oil bath after the completion of the dropwise addition, the reflux reaction is carried out for 10 hours (the reaction process adopts TLC tracking detection, the developing agent is a petroleum ether and ethyl acetate mixed solution with the volume ratio of 1:1), the solvent is distilled off from the reaction liquid, the residue is subjected to column chromatography, namely, the residue after the solvent is distilled off is taken, 10mL of petroleum ether solvent is added to dissolve the residue to obtain a dissolving liquid, then 1.5 g of silica gel (300-400-mesh coarse pore (zcx.II) column chromatography silica gel) is added into the dissolving liquid, after the solvent is distilled off, a mixture of the dried residue and the silica gel is obtained, the mixture is packed into a column with the volume ratio of 1:1 as eluent, TLC tracking detection (developing solvent is mixed solution of petroleum ether and ethyl acetate in volume ratio of 1:1), collecting eluent containing a compound shown in formula (IB-5) according to TLC detection, evaporating the solvent from the collected eluent, and drying to obtain a white solid product, namely the compound (IB-5), wherein the yield is 48% (based on the amount of 2, 4-dichloroquinazoline (II)) and the melting point is 269-271 ℃. 1 HNMR(500MHz,DMSO-d 6 )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).
Example 23: in vitro test of anticancer Activity
The prepared compounds (IA-1) to (IA-9) are respectively subjected to biological activity tests of human breast cancer cell lines MDA-MB-231, human liver cancer cells Huh7 and human liver cancer cells HepG2, wherein the compound (IA-1) is prepared by the method of the example 3. The prepared compounds (IB-1) to (IB-5) are respectively subjected to biological activity tests of human breast cancer cell lines MDA-MB-231, human liver cancer cells Huh7 and human liver cancer cells HepG2, wherein the compound (IB-1) is prepared by the method of example 15.
The testing method comprises the following steps: tetrazolium salt reduction method (MTT method).
Cell lines: human breast cancer cell strain MDA-MB-231, human liver cancer cell Huh7 and human liver cancer cell HepG2. The cell lines are purchased from cell banks of Shanghai Seisakusho of China academy of sciences.
The experimental procedure was as follows:
(1) Culture of tumor cells
The cell algebra used in the experiment is within 5 generations. The consumables and reagents used for cell culture were subjected to a strict sterilization procedure, and all experimental procedures were performed in a sterile operating table.
(a) Resuscitation of tumor cells
Before the experiment starts, articles to be used, such as a culture bottle, a centrifuge tube, a pipette, a gun head, a waste liquid barrel and the like, are placed into an ultra-clean bench to be sterilized by turning on an ultraviolet lamp, and reagents used for cell culture are placed into a constant-temperature water bath kettle at 37 ℃ to be preheated. All the work is ready, the ultraviolet lamp is turned off, and the super clean bench fluorescent lamp and the ventilating fan are turned on. Taking out frozen tumor cells from a refrigerator at the temperature of minus 80 ℃, shaking in a water bath at the temperature of 37 ℃ to quickly defrost, spraying alcohol into an ultra-clean bench under the condition that a small part of frozen liquid in a frozen tube is still unfrozen, immediately transferring all the cells in the frozen tube into a 15mL centrifuge tube added with 1640 culture solution, and lightly blowing and uniformly mixing by a pipetting gun. The centrifuge tube was placed in a centrifuge and centrifuged at 1000rpm for 5min, and the supernatant was aspirated off in a super clean bench. Sucking 2mL1640 culture solution into a centrifuge tube containing cell sediment by a pipette to prepare a cell suspension, blowing and uniformly mixing the cell suspension, and transferring the cell suspension to a bottle bottom with an area of 25cm 2 4mL of 1640 culture solution is added into the breathable culture flask, the culture flask is gently shaken to mix cells, and the culture flask is put into 5% CO 2 Culturing in a constant temperature incubator at 37 ℃.
(b) Passage of tumor cells
When the cell growth state is good and 70% -80% of the bottom of the culture flask is paved, the cell can be passaged. The steps of the passage operation are all completed in an ultra-clean bench, and the passage can be started after the preparation work is finished. Firstly, sucking the original culture solution in a culture bottle into a waste liquid barrel by a liquid transferring gun, adding 2mL of PBS, repeatedly washing for several times, sucking, then adding about 600-700 mu L of trypsin (containing 0.02% EDTA, phenol red and 0.25% pancreatin), ensuring that the bottle bottom is completely covered by trypsin, slightly shaking the culture bottle, placing the culture bottle into a constant temperature incubator at 37 ℃ for incubation for 1-2 min, observing the cell shedding condition under a microscope, and if a small number of cells still adhere to the wall, lightly beating the wall of the bottle by using a finger belly until most of the cells can shed from the bottle bottom. After the end of digestion by adding 2mL of 1640 culture solution, the cells were gently blown off with a pipette, the cells were all blown off from the bottom of the flask, and the cell suspension was transferred to a 15mL centrifuge tube, 1000rpm, and centrifuged for 5min. Sucking the supernatant, diluting with 1640 culture solution, blowing uniformly, and evenly distributing into 2-3 culture bottles to continue in 5% CO 2 Culturing in a constant temperature incubator at 37 ℃.
(c) Cryopreservation of tumor cells
Cell cryopreservation solutions were prepared in advance according to the volume ratio of FBS (fetal bovine serum): dmso=9:1 and placed in a refrigerator at 4 ℃ for later use. The freezing operation steps are completed in the super clean bench, and the frozen storage can be started after the preparation work is finished. Firstly, sucking the original culture solution in a culture flask, adding 2mL of PBS buffer solution, repeatedly washing for several times, pouring, then adding about 600-700 mu L of trypsin (containing 0.02% EDTA, phenol red and 0.25% pancreatin), gently shaking the culture flask, ensuring that the pancreatin can cover the bottom of the culture flask, placing the culture flask into a constant-temperature incubator at 37 ℃ for incubation for 1-2 min, observing the cell shedding condition under a microscope, if a small number of cells still cling to the wall, lightly beating the wall of the bottle by using a finger belly until the cells can be mostly shed from the bottom of the bottle. After the end of digestion by adding 2mL of 1640 culture solution, the cells were gently blown off with a pipette, the cells were all blown off from the bottom of the flask, and the cell suspension was transferred to a 15mL centrifuge tube, 1000rpm, and centrifuged for 5min. The supernatant was decanted. 1mL of frozen stock solution just taken out from a refrigerator at 4 ℃ is added, and is blown to be uniform to form cell suspension, and the cell suspension is transferred into a frozen stock tube. Placing the labeled cell types, cell algebra and frozen date into a refrigerator with ultralow temperature of-80 ℃ for preservation after placing the labeled cell types, cell algebra and frozen date into the refrigerator with ultralow temperature of-20 ℃ for standing for 1 hour at-4 ℃.
(2) MTT experimental method
(a) Cell count: after digesting and centrifuging tumor cells with good growth state in a culture flask, re-suspending the tumor cells with 4mL1640 culture solution, taking 10 mu L of cell suspension to a cell counting plate, and diluting B16F10 cells to 5 multiplied by 10 after counting 4 And each mL.
(b) And (3) paving: taking 96-well plate, adding 100 μl diluted cell suspension into experimental well, adding 100 μl1640 culture solution into blank well, adding 100 μl LPBS buffer around, adding CO 2 Culturing in a constant temperature incubator.
(c) Compounding and adding a compound: the compound 10. Mu. Mol/mL prepared in DMSO and the positive control Sorafenib were diluted to the indicated concentrations in 1640 medium, and the drug was applied by the liquid exchange method at 40. Mu.M, 20. Mu.M, 10. Mu.M, 5. Mu.M, 2.5. Mu.M, and 1.25. Mu.M, respectively. Discarding original culture solution, adding 100 μl of 1640 culture solution containing compound or positive control drug into experimental hole, adding 100 μl of 1640 culture solution into control group and blank hole, adding 96-well plate into 5% CO after adding drug 2 Continuously culturing in a constant temperature incubator at 37 ℃.
(d) Adding MTT: after 48 hours, the 96-well plate is taken out and put into an ultra clean bench, 10 mu L of MTT solution with concentration of 5mg/mL is added into each well under the condition of light shielding, and then 5% CO is added 2 Continuously culturing in a constant temperature incubator at 37 ℃.
(e) And (3) detection: incubating the MTT-added 96-well plate in an incubator for 3.5-4 h, taking out, carefully sucking out the solution in each well, adding 150 mu L of DMSO into each well to dissolve formazan generated by dissolution, then placing the formazan on a flat-plate oscillator to oscillate for 20min, and detecting the absorbance value at 490nm by using an enzyme-labeled instrument.
(f) Experimental data processing: the cell viability was calculated according to the following formula, and the value of 50% of the cell viability was IC 50 。
Cell viability (%) = [ (As-Ab)/(Ac-Ab) ] ×100%
As: experiment well (cell-containing culture solution, MTT, toxic substance (i.e., compound (IA) or (IB))
Ac control wells (cell-containing culture medium, MTT, without toxic substances)
Ab-blank wells (MTT-containing, cell and toxic material free culture broth).
The results of the test are shown in table 1:
TABLE 1 inhibition of cancer cell growth by Compound (IA) (IC 50 ，μΜ)
Compounds of formula (I) 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 (Sorafenib) 9.14±0.82 8.15±0.26 -
TABLE 2 inhibition of cancer cell growth by Compound (IB) (IC 50 ，μΜ)
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Claims (6)

1. A phenylureido quinazoline compound, characterized in that: the structure of the phenylureido quinazoline compound is shown as a formula (IA) or (IB):
in the formula (IA), R is H, 4-methoxy, 2, 5-difluoro, or 3-methyl; in the formula (IB), R is H, 3-trifluoromethyl-4-chloro, or 3-methoxy.
2. A process for the preparation of phenylureido quinazolines according to claim 1, wherein: the preparation method comprises the following steps: (1) Adding a compound shown in a formula (II) into an organic solvent A, stirring and dissolving, dropwise adding a mixed solution containing the compound shown in a formula (IIIA) or (IIIB), a catalyst B and the organic solvent A under the condition of stirring at room temperature, heating the reaction solution to a reflux state after the dripping is finished, stirring and reacting for 0.5-12 hours, and separating and purifying the reaction solution to obtain phenylureido quinazoline compounds shown in a formula (IA) or (IB);
the organic solvent A is one of the following: dichloromethane, ethanol, isopropanol or toluene;
the catalyst B is one of the following: triethylamine, 4-dimethylaminopyridine, pyridine or sodium hydroxide;
in the formula (IIIA), R is defined as in the formula (IA); in formula (IIIB), R is as defined in formula (IB).
3. The method of manufacturing as claimed in claim 2, wherein: the ratio of the amount of the compound represented by the formula (II), the compound represented by the formula (IIIA) or (IIIB) to the amount of the catalyst B to be fed is 1:0.8 to 1.2:0.1 to 1.
4. The use of a phenylureido quinazoline compound according to claim 1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the prophylaxis or treatment of liver cancer.
5. The use according to claim 4, wherein: the medicine for preventing or treating liver cancer is a medicine for preventing or treating human liver cancer cell HepG2.
6. The use according to claim 4, wherein: the medicine for preventing or treating liver cancer is a medicine for preventing or treating human liver cancer cell Huh 7; in the structural formula (IA) of the phenylureido quinazoline compound, R is 2, 5-difluoro; in the formula (IB), R is 3-trifluoromethyl-4-chlorine or 3-methoxy.
CN202311388524.8A 2023-10-25 2023-10-25 Phenyl ureido quinazoline compound, preparation method thereof and application thereof in preparation of medicines for preventing or treating liver cancer Pending CN117800925A (en)

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