CN108042546B - Application of morpholinyl acetamidobenzo [ d ] aza-based quinazoline compound in preparation of drugs for treating cervical cancer - Google Patents

Abstract

基喹唑啉类化合物在制备预防或治疗肿瘤药物中的应用，特别是制备预防或治疗人宫颈癌药物中的应用，其对抑制人宫颈癌细胞株Siha活性具有显著效果。The invention discloses a morpholinoacetamidobenzo[d]azepine

The application of the quinazoline compound in the preparation of a medicament for preventing or treating tumors, especially in the preparation of a medicament for preventing or treating human cervical cancer, has a significant effect on inhibiting the activity of the human cervical cancer cell line Siha.

Description

Morpholinylacetamidobenzo[d]aza*ylquinazoline compounds in the preparation of therapeutic Application of drugs in the treatment of cervical cancer

(1) Technical field

基喹唑啉类化合物在制备预防或治疗人宫颈癌药物中的应用。The invention relates to a quinazoline compound and application thereof, in particular to a morpholinoacetamidobenzo[d]azepine

The application of quinazoline compounds in the preparation of medicaments for preventing or treating human cervical cancer.

(2) Background technology

Quinazoline compounds have many good biological activities and are widely used in the field of medicine, especially some quinazoline derivatives with special structures have obvious antiviral activity, antibacterial activity, antitumor activity, etc. Some types of compounds have been marketed as antitumor drugs. For example, Gefitinib and Erlotinib, which are marketed for the treatment of lung cancer, and Lapatinib, which are used for the treatment of breast cancer, are all quinazoline compounds. Novel quinazoline compounds and their biological activities are also commonly 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, 1189-1204). Of course, most quinazoline compounds do not have antitumor activity.

(3) Contents of the invention

基喹唑啉类化合物的应用，该类化合物在一定剂量下对人宫颈癌细胞株Siha具有很好的抑制效果；且该类化合物制备方法简便，易于操作，原料易得，且生产成本较低，适于工业化应用。The object of the present invention is to provide a kind of novel quinazoline compound - morpholino acetamidobenzo [d] aza

The application of quinazoline compounds, the compounds have a good inhibitory effect on the human cervical cancer cell line Siha under a certain dose; and the preparation method of the compounds is simple, easy to operate, easy to obtain raw materials, and low production cost , suitable for industrial applications.

For realizing the above-mentioned purpose of the invention, the present invention adopts following technical scheme:

基喹唑啉类化合物在制备预防或治疗肿瘤药物中的应用，特别是在制备预防或治疗人宫颈癌药物中的应用：The present invention provides a morpholinoacetamidobenzo[d]azepine represented by formula (I)

The application of quinazoline compounds in the preparation of drugs for preventing or treating tumors, especially the application in preparing drugs for preventing or treating human cervical cancer:

Preferably, the drug is a drug with the activity of inhibiting human cervical cancer cell line Siha.

基喹唑啉类化合物的制备方法，所述的方法为：(1)将式(Ⅱ)所示化合物与式(Ⅲ)所示化合物混合，在有机溶剂A中，于碱性催化剂B的作用下，25～120℃进行反应(TLC跟踪监测，展开剂为乙酸乙酯/石油醚＝1：3(v/v)，优选40～100℃反应0.5～12h)，反应完全后，将反应液分离纯化，制得式(Ⅳ)所示化合物；所述有机溶剂A选自下列之一：氯仿、甲苯、甲醇、乙醇、丙醇、异丙醇、乙腈或N,N-二甲基甲酰胺；所述的碱性催化剂B选自下列之一：吡啶、二乙胺、三乙胺、喹啉、N,N-二甲苯胺、4-二甲氨基吡啶、4-吡咯烷基吡啶或碳酸钠(优选吡啶、二乙胺、三乙胺、N,N-二甲苯胺或4-二甲氨基吡啶)；In addition, the present invention provides a morpholinoacetamidobenzo[d]azepine represented by formula (I)

The preparation method of quinazoline compounds, the method is as follows: (1) the compound represented by formula (II) is mixed with the compound represented by formula (III), in organic solvent A, under the action of basic catalyst B at 25～120℃ (TLC tracking monitoring, the developing solvent is ethyl acetate/petroleum ether=1:3 (v/v), preferably 40～100℃ for 0.5～12h), after the reaction is complete, the reaction solution separation and purification to obtain the compound represented by formula (IV); the organic solvent A is selected from one of the following: chloroform, toluene, methanol, ethanol, propanol, isopropanol, acetonitrile or N,N-dimethylformamide ; Described basic catalyst B is selected from one of the following: pyridine, diethylamine, triethylamine, quinoline, N,N-dimethylaminopyridine, 4-dimethylaminopyridine, 4-pyrrolidinopyridine or carbonic acid Sodium (preferably pyridine, diethylamine, triethylamine, N,N-dimethylaminopyridine or 4-dimethylaminopyridine);

(2) The compound represented by the formula (IV) is in the organic solvent D, under the action of the reducing agent E, the reaction is complete at 25～100 ℃ (TLC tracking monitoring, the developing solvent is ethyl acetate/petroleum ether=1:1 ( v/v), preferably at 40-80°C for 0.5-12h), the reaction solution is filtered, and the concentrate after the filtrate is concentrated under reduced pressure is dried (preferably at 25°C under vacuum) to obtain the compound represented by formula (V); the The organic solvent D is one of the following: chloroform, toluene, methanol, ethanol, propanol, isopropanol, acetonitrile or N,N-dimethylformamide; the reducing agent E is one of the following: iron powder/concentrated hydrochloric acid , iron powder/acetic acid, palladium carbon/ammonium formate or palladium carbon/hydrazine hydrate; the iron powder/concentrated hydrochloric acid refers to the mixture of iron powder and concentrated hydrochloric acid in any proportion, and iron powder/acetic acid refers to the mixture of iron powder and acetic acid in any proportion Mixing, described palladium carbon/ammonium formate refers to the mixing of palladium carbon and ammonium formate in any proportion, and described palladium carbon/hydrazine hydrate is the mixture of palladium carbon and hydrazine hydrate in any proportion;

(3) Mix the compound represented by formula (V) with chloroacetyl chloride or chloroacetic anhydride, under the action of basic catalyst F, in organic solvent G, the reaction is complete at -10～50℃ (TLC tracking monitoring, the developing agent is Ethyl acetate/petroleum ether=1:1 (v/v), preferably -10～50℃ for 3～12h), the reaction solution is subjected to post-treatment A to obtain the compound represented by formula (VI); the base The catalyst F is one of the following: pyridine, diethylamine, triethylamine, quinoline, N,N-dimethylaminopyridine, 4-dimethylaminopyridine, 4-pyrrolidinopyridine or sodium carbonate; the organic solvent G is one of the following: tetrahydrofuran, dichloromethane, chloroform, ethyl acetate, diethyl ether, acetonitrile, toluene or benzene;

(4) Mix the compound represented by formula (VI) with morpholine, in organic solvent J, under the action of basic catalyst K, carry out reaction at 25～120 ℃ (TLC tracking monitoring, developing solvent is ethyl acetate/petroleum ether=1:1 (v/v), preferably 40～100℃ for 0.5～36h), after the reaction is complete, the reaction solution is subjected to post-treatment B to obtain the compound represented by formula (I); the organic solvent J is selected from From one of the following: chloroform, toluene, methanol, ethanol, propanol, isopropanol, acetonitrile or N,N-dimethylformamide; the basic catalyst K is selected from one of the following: pyridine, triethylamine , quinoline, N,N-xylidine, 4-dimethylaminopyridine, 4-pyrrolidinopyridine or sodium carbonate (preferably pyridine, quinoline, triethylamine, N,N-xylidine or 4-ditoluidine methylaminopyridine).

Further, in step (1), the ratio of the amount of the compound represented by the formula (III) to the compound represented by the formula (II) and the amount of the basic catalyst B charged is 1.0:0.8-1.2:1.0-8.0.

Further, in step (1), the amount of the organic solvent A used is 10-50 mL/g in terms of the mass of the compound represented by formula (III).

Further, the method for separating and purifying the reaction solution described in the step (1) of the present invention is as follows: after the reaction is complete, the reaction solution is evaporated to remove the solvent, and the concentrate is dissolved in an organic solvent C to obtain a solution, and then add the solution to the solution. Add 1.0 to 2.0 times the weight of the concentrate with column chromatography silica gel, and after mixing, evaporate the solvent and dry to obtain a mixture of the concentrate and silica gel. The mixed solution with ethyl acetate is the eluent, and the effluent containing the target component is collected (preferably with ethyl acetate/petroleum ether=1:3 (v/v) as the developing agent for tracking detection, and the target component is collected, preferably components with an Rf value of 0.5), concentrated under reduced pressure, and dried (preferably at 50° C.) to obtain the compound represented by formula (IV); the organic solvent C is one of the following: ethanol, chloroform, tetrahydrofuran or ethyl acetate . The amount of the organic solvent C can be used to dissolve the residue.

Further, in step (2), when described reducing agent E is iron powder/concentrated hydrochloric acid or iron powder/acetic acid, the compound shown in formula (IV) and the iron powder in reducing agent E, concentrated hydrochloric acid or acetic acid are mixed. The feeding mass ratio is 1.0:1.0～3.0:0.2～1.0. In the present invention, the mass concentration of concentrated hydrochloric acid is 36% to 38%, and the acetic acid adopts glacial acetic acid.

Further, in step (2), when described reducing agent E is palladium carbon/ammonium formate or palladium carbon/hydrazine hydrate, the compound shown in formula (IV) and palladium carbon, ammonium formate or hydrazine in reducing agent E The feeding mass ratio of hydrazine is 1.0:0.1-0.5:1.0-3.0. The mass loading of palladium in the palladium carbon applicable in the present invention is 2-10%, preferably 5%, and the mass concentration of hydrazine hydrate is 40-80%, preferably 80%.

Further, in step (2), the amount of the organic solvent D used is 10-50 mL/g in terms of the mass of the compound represented by formula (IV).

Further, in step (3), the ratio of the amount of the compound represented by the formula (V) to chloroacetyl chloride or chloroacetic anhydride and basic catalyst F is 1:1.0-8.0:1.0-3.0.

Further, in step (3), the amount of the organic solvent G used is 11-100 mL/g in terms of the mass of the compound represented by formula (V).

Further, the present invention specifically recommends that step (3) be carried out according to the following method: under the condition of -10～10℃, add the compound represented by formula (V) and the organic solvent G solution of basic catalyst F or the solution of formula (V) in organic solvent G The organic solvent G solution of chloroacetyl chloride or chloroacetic anhydride is added dropwise to the shown compound and the basic catalyst F, and after the dropping is completed, the reaction is carried out at -10 to 50 °C for 3 to 12 hours, and the obtained reaction solution is subjected to post-treatment A to obtain the formula (VI). The volumetric dosage of the organic solvent for dissolving chloroacetyl chloride or chloroacetic anhydride has no effect on the present invention, and the total dosage of the organic solvent G is 11-100 mL/g in terms of the mass of the compound represented by formula (V). The total amount of the organic solvent G refers to the total volume of the organic solvent G for dissolving the basic catalyst F and the compound represented by formula (V) and the organic solvent G for dissolving chloroacetyl chloride or chloroacetic anhydride.

Further, the method for separating and purifying the reaction solution described in step (3) of the present invention is as follows: after the reaction is completed, the reaction solution is filtered, the filtrate is evaporated to remove the solvent, and the concentrate is dissolved with an organic solvent H to obtain a solution, and then the solution is added to the solution. Add 1.0 to 2.0 times the weight of column chromatography silica gel of the concentrate to the liquid, and after mixing, evaporate the solvent and dry to obtain a mixture of the concentrate and silica gel. The mixed solution of petroleum ether and ethyl acetate is the eluent, and the effluent containing the target component is collected (preferably with ethyl acetate/petroleum ether=1:1 (v/v) as the developing agent for tracking detection, and the target component is collected, It is preferable to collect components with an Rf value of 0.5), concentrate under reduced pressure, and dry (preferably at 50°C) to obtain the compound represented by formula (VI); the organic solvent H is one of the following: ethanol, chloroform, tetrahydrofuran or acetic acid ethyl ester. The amount of the organic solvent H can be used to dissolve the residue.

Further, in step (4), the ratio of the amount of the compound represented by the formula (VI) to morpholine and the amount of the basic catalyst K is 1.0:0.8-8.0:1.0-8.0.

Further, in step (4), the amount of the organic solvent J used is 10-60 mL/g in terms of the mass of the compound represented by formula (VI).

Further, the method for the post-treatment B of the reaction solution described in the step (4) of the present invention is: after the reaction is complete, the reaction solution is evaporated to remove the solvent, and the concentrate is dissolved in an organic solvent M to obtain a solution, and then add the solution to the solution. Add 1.0 to 2.0 times the weight of the concentrate to column chromatography silica gel, and after mixing, evaporate the solvent and dry to obtain a mixture of the concentrate and silica gel. The mixed solution of ether and ethyl acetate is used as the eluent, and the effluent containing the target component is collected (preferably with ethyl acetate/petroleum ether=1:1 (v/v) as the developing agent for tracking detection, and the target component is collected, preferably The components with Rf value of 0.5 are collected), concentrated under reduced pressure, and dried (preferably at 50°C) to obtain the compound represented by formula (I); the organic solvent M is one of the following: ethanol, chloroform, tetrahydrofuran or ethyl acetate ester. The amount of the organic solvent M is sufficient to dissolve the residue.

The organic solvents A, C, D, G, H, J and M of the present invention are all organic solvents, and are named for the convenience of distinguishing different organic solvents used in different steps, and the letters themselves have no meaning; the catalyst B, reducing agent E, Catalyst F and catalyst K are both catalysts, and are named for the convenience of distinguishing different catalysts used in different steps, and the letters themselves have no meaning; the post-treatment A and post-treatment B are both post-treatments, and are named for the convenience of distinguishing different post-treatments used in different steps. , the letters themselves have no meaning.

基喹唑啉类化合物(Ⅰ)在制备预防或治疗人宫颈癌药物中的应用，该化合物对人宫颈癌细胞株Siha具有显著的抑制活性。The beneficial effects of the present invention are mainly reflected in: providing a novel quinazoline compound—morpholinylacetamidobenzo[d]azepine

The application of the quinazoline compound (I) in the preparation of a medicament for preventing or treating human cervical cancer, the compound has significant inhibitory activity on the human cervical cancer cell line Siha.

(4) Specific implementations

The present invention will be further described with reference to specific embodiments. The following embodiments are intended to illustrate the present invention, but not to limit the present invention in any way.

The preparation of compound (II) was obtained by referring to the method of literature (Weinstock, J. et al. J. Med. Chem., 1986, 29(11), 2315-2325). The preparation of 4-chloro-6-nitroquinazoline (III) was obtained by referring to the method in the literature (Fernandes, C. etal. Bioorg. Med. Chem., 2007, 15(12), 3974-3980).

The palladium carbon (Pd/C) model D5H5A used in the examples of the present invention was purchased from Shaanxi Ruike New Materials Co., Ltd.

Example 1: Preparation of 6-nitroquinazoline (IV)

1.20 g (5.73 mmol) of 4-chloro-6-nitroquinazoline (III) and 2.39 g (6.87 mmol) of compound (II), 3.62 g (45.76 mmol) of pyridine, and 12 mL of chloroform were added to 50 mL of reaction In the bottle, heated to 40°C, followed by TLC detection (developing solvent is ethyl acetate/petroleum ether=1:3 (v/v)), stirred and reacted for 10 hours, closed the reaction, evaporated the solvent from the reaction solution, and obtained the concentrate Add 10 ml of ethyl acetate to dissolve it to obtain a dissolving solution, add 3.0 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the dissolving solution, and after mixing, evaporate the solvent to obtain a dry concentrate The mixture with silica gel, pack the mixture into a column, and then use the petroleum ether/ethyl acetate mixed solution with a volume ratio of 1:10 as the eluent, elution, and TLC tracking detection (developing solvent is ethyl acetate/petroleum ether=1 : 3(v/v)), according to TLC detection, collect the eluate containing the compound shown in formula (IV) (Rf value is 0.5), the collected solution is concentrated, and dried at 50 ° C to obtain the pale yellow shown in formula (IV) Solid product, yield 85.1%, melting point 164～166℃. ¹ H NMR (500 MHz, CDCl ₃ ) δ: 3.32-3.38 (m, 1H), 3.63 (dt, J=3.4, 15.5 Hz, 1H), 3.75 (s, 3H), 3.82 (s, 6H), 3.91 ( dd,J＝8.1,14.3Hz,1H),4.03(td,J＝4.1,11.7Hz,1H),4.15(d,J＝11.5Hz,1H),4.72(dd,J＝8.3,14.2Hz,1H) ), 5.14(t, J＝8.9Hz, 1H), 6.60(s, 1H), 6.90(d, J＝8.7Hz, 2H), 7.08(d, J＝8.6Hz, 2H), 7.93(d, J = 9.1 Hz, 1H), 8.48 (dd, J=2.4, 9.2 Hz, 1H), 8.71 (s, 1H), 8.96 (d, J=2.4 Hz, 1H). IR(KBr,cm ^-1 )ν: 2917, 2848, 1616, 1580, 1510, 1463, 1355, 1327, 1249, 1038,847.

Example 2: Preparation of 6-nitroquinazoline (IV)

1.20 g (5.73 mmol) of 4-chloro-6-nitroquinazoline (III) and 1.59 g (4.57 mmol) of compound (II), 1.67 g (22.83 mmol) of diethylamine, and 60 mL of toluene were added to 100 mL of In the three-necked flask, heated to 100 ° C, TLC tracking detection (developing solvent is ethyl acetate/petroleum ether=1:3 (v/v)), stirred and reacted for 2 hours, closed the reaction, the reaction solution was evaporated to remove the solvent, and the obtained Add 20 ml of ethanol to the concentrate to dissolve it to obtain a dissolving solution, add 2.5 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the dissolving solution, and after mixing, evaporate the solvent to obtain a dry concentrate The mixture with silica gel, the mixture is packed into a column, and then use the petroleum ether/ethyl acetate mixed solution with a volume ratio of 1:5 as the eluent, elution, and TLC tracking detection (developing solvent is ethyl acetate/petroleum ether=1 : 3(v/v)), according to TLC detection, collect the eluate containing the compound shown in formula (IV) (Rf value is 0.5), the collected solution is concentrated, and dried at 50 ° C to obtain the pale yellow shown in formula (IV) Solid product, yield 72.6%, melting point 164～166℃. ¹ H NMR and IR were the same as in Example 1.

Example 3: Preparation of 6-nitroquinazoline (IV)

1.20 g (5.73 mmol) of 4-chloro-6-nitroquinazoline (III) and 1.99 g (5.72 mmol) of compound (II), 0.58 g (5.73 mmol) of triethylamine and 60 mL of ethanol were added to 100 mL of In the three-necked flask, heated to 60 ° C, TLC tracking detection (developing solvent is ethyl acetate/petroleum ether=1:3 (v/v)), stirred for 8 hours, closed the reaction, the reaction solution was evaporated to remove the solvent, and the obtained Add 20 ml of chloroform to the concentrate to dissolve it to obtain a dissolving solution, add 2.5 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the dissolving solution, and after mixing, evaporate the solvent to obtain a dry concentrate The mixture with silica gel, pack the mixture into a column, and then use the petroleum ether/ethyl acetate mixed solution with a volume ratio of 10:1 as the eluent, elution, and TLC tracking detection (developing solvent is ethyl acetate/petroleum ether=1 : 3(v/v)), according to TLC detection, collect the eluate containing the compound shown in formula (IV) (Rf value is 0.5), the collected solution is concentrated, and dried at 50 ° C to obtain the pale yellow shown in formula (IV) Solid product, yield 77.2%, melting point 164-166°C. ¹ H NMR and IR were the same as in Example 1.

Example 4: Preparation of 6-nitroquinazoline (IV)

1.20 g (5.73 mmol) of 4-chloro-6-nitroquinazoline (III) and 2.20 g (6.32 mmol) of compound (II), 1.40 g (11.46 mmol) of 4-dimethylaminopyridine, 60 mL of iso- Propanol was added to the three-necked flask of 100 ml, stirred at room temperature of 25° C., followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:3 (v/v)), reacted for 12 hours, closed the reaction, and evaporated the reaction solution. Solvent, add 20 ml of tetrahydrofuran to the obtained concentrate to dissolve it to obtain a dissolving solution, add 4.0 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the dissolving solution, and after mixing, evaporate the solvent to obtain The mixture of the dried concentrate and silica gel was packed into a column, and then a 5:1 petroleum ether/ethyl acetate mixed solution by volume was used as the eluent to elute, and TLC tracking detection (developing solvent was ethyl acetate/ethyl acetate/ethyl acetate) was used as the eluent. Petroleum ether=1:3 (v/v)), according to TLC detection, collect the eluate containing the compound shown in formula (IV) (Rf value is 0.5), concentrate the collected solution, and dry at 50 °C to obtain the compound of formula (IV). The light yellow solid product is shown, the yield is 80.2%, and the melting point is 164-166°C. ¹ H NMR and IR were the same as in Example 1.

Example 5: Preparation of 6-nitroquinazoline (IV)

1.20 g (5.73 mmol) of 4-chloro-6-nitroquinazoline (III) and 1.79 g (5.15 mmol) of compound (II), 1.04 g (8.58 mmol) of N,N-xylidine, 12 mL of N,N-dimethylformamide was added to a 50 ml reaction flask, heated to 120°C, followed by TLC detection (developing solvent: ethyl acetate/petroleum ether=1:3 (v/v)), and the reaction was stirred for 0.5 hours , the reaction was closed, the solvent was evaporated from the reaction solution, 20 ml of tetrahydrofuran was added to the obtained concentrate to dissolve it to obtain a dissolving solution, 5.0 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) was added to the dissolving solution, and the mixture was mixed. After homogenization, the solvent was evaporated to obtain a mixture of dry concentrate and silica gel. The mixture was packed into a column, and then a 1:1 volume ratio of petroleum ether/ethyl acetate mixed solution was used as the eluent for elution, followed by TLC tracking detection. (developing solvent is ethyl acetate/petroleum ether=1:3 (v/v)), collect the eluate containing the compound represented by formula (IV) according to TLC detection (Rf value is 0.5), the collected solution is concentrated, 50 After drying at °C, a pale yellow solid product represented by formula (IV) was obtained, the yield was 89.6%, and the melting point was 164-166 °C. ¹ H NMR and IR were the same as in Example 1.

Example 6: Preparation of 6-nitroquinazoline (IV)

1.20 g (5.73 mmol) of 4-chloro-6-nitroquinazoline (III) and 2.39 g (6.87 mmol) of compound (II), 3.62 g (45.76 mmol) of pyridine, and 20 mL of propanol were added to 50 mL of In the reaction flask, heated to 40°C, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:3 (v/v)), stirred and reacted for 10 hours, closed the reaction, evaporated the solvent from the reaction solution, and obtained concentrated Add 20 ml of ethyl acetate to dissolve it to obtain a dissolving solution, add 3.5 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the dissolving solution, and after mixing, evaporate the solvent to obtain a dry concentrate The mixture of the compound and silica gel, the mixture was packed into a column, and then the petroleum ether/ethyl acetate mixed solution with a volume ratio of 1:1 was used as the eluent, and the elution was carried out by TLC tracking detection (developing solvent was ethyl acetate/petroleum ether= 1:3 (v/v)), according to TLC detection, collect the eluate containing the compound shown in formula (IV) (Rf value is 0.5), the collected solution is concentrated, and dried at 50 °C to obtain the pale compound shown in formula (IV). Yellow solid product, yield 78.3%, melting point 164-166°C. ¹ H NMR and IR were the same as in Example 1.

Example 7: Preparation of 6-aminoquinazoline (V)

0.40 g (0.77 mmol) of 6-nitroquinazoline (IV) prepared by the method of Example 1, 0.40 g (6.34 mmol) of ammonium formate, 0.04 g of 5% Pd/C, and 4.0 ml of chloroform were successively added to the reaction flask , stirred at room temperature at 25°C, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1 (v/v)), reacted for 12 hours, filtered, concentrated the filtrate, and vacuum-dried at 25°C to obtain a pale yellow solid product 6- Aminoquinazoline (V), yield 98.2%, melting point 122～126℃. ¹ H NMR (500MHz, CDCl ₃ )δ: 3.40-3.48(m, 2H), 3.71(s, 3H), 3.82(s, 3H), 3.83(s, 3H), 3.87-3.98(m, 5H), 4.45(dd,J＝6.3,13.8Hz,1H),4.95(dd,J＝6.5,9.2Hz,1H),6.47(s,1H),6.90(d,J＝8.7Hz,2H),6.95(d , J＝2.5Hz, 1H), 7.11(d, J＝8.6Hz, 2H), 7.15(dd, J＝8.9, 2.5Hz, 1H), 7.69(d, J＝8.9Hz, 1H), 8.50(s , 1H). IR(KBr,cm ^-1 )ν: 3368, 3215, 2932, 2825, 1628, 1566, 1512, 1487, 1353, 1248, 1036,834.

Example 8: Preparation of 6-aminoquinazoline (V)

0.40 g (0.77 mmol) of 6-nitroquinazoline (IV) prepared by the method of Example 2, 1.20 g (19.18 mmol) of 80wt% hydrazine hydrate, 0.20 g of 5% Pd/C, and 20.0 ml of toluene were added to 50 in turn. The reaction flask was heated to 100°C, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1 (v/v)), stirred and reacted for 0.5 hours, cooled and filtered, the filtrate was concentrated, and vacuum-dried at 25°C A pale yellow solid product, 6-aminoquinazoline (V), was obtained with a yield of 100.0% and a melting point of 122-126°C. ¹ H NMR and IR were the same as in Example 7.

Example 9: Preparation of 6-aminoquinazoline (V)

0.40 g (0.77 mmol) 6-nitroquinazoline (IV) prepared by the method of Example 3, 0.08 g concentrated hydrochloric acid (mass concentration 36-38%), 0.40 g iron powder, 20.0 ml methanol were added to 50 ml successively. The reaction flask was heated to 40°C, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1 (v/v)), stirred and reacted for 8 hours, cooled and filtered, the filtrate was concentrated, and vacuum-dried at 25°C to obtain Light yellow solid product 6-aminoquinazoline (V), yield 94.1%, melting point 122～126℃. ¹ H NMR and IR were the same as in Example 7.

Example 10: Preparation of 6-aminoquinazoline (V)

0.40 g (0.77 mmol) of 6-nitroquinazoline (IV) prepared by the method of Example 4, 0.40 g of acetic acid, 1.20 g of iron powder, and 20.0 ml of isopropanol were successively added to a 50 ml reaction flask, and heated to 80°C, TLC tracking detection (developing solvent is ethyl acetate/petroleum ether=1:1 (v/v)), stirring and reacting for 3 hours, cooling and filtration, the filtrate is concentrated, and vacuum-dried at 25°C to obtain a pale yellow solid product 6-amino Quinazoline (V), yield 97.5%, melting point 122～126℃. ¹ H NMR and IR were the same as in Example 7.

Example 11: Preparation of chloroacetamidoquinazoline (VI)

0.27 g (0.55 mmol) of 6-aminoquinazoline (V) prepared by the method of Example 7, 0.13 g (1.64 mmol) of pyridine, and 3 ml of tetrahydrofuran were successively added to the reaction flask, and 0.497 g of tetrahydrofuran was added dropwise with stirring at -10°C. g (4.40mmol) chloroacetyl chloride, dripped, TLC tracking detection (developing solvent is ethyl acetate/petroleum ether=1:1), reacted under -10 ℃ for 12 hours, filtered, the filtrate was evaporated to remove the solvent, and the concentrate was added Dissolve it in 10 ml of ethyl acetate to obtain a dissolving solution. Add 0.60 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the dissolving solution. After mixing, the solvent is evaporated to obtain a dry concentrate and silica gel. The mixture was packed into a column, and then the petroleum ether/ethyl acetate mixed solution with a volume ratio of 1:10 was used as the eluent, and the elution was carried out by TLC tracking detection (developing solvent was ethyl acetate/petroleum ether=1:1 (v/v)), according to TLC detection, collect the eluate containing the compound shown in formula (VI) (Rf value is 0.5), the collected solution is concentrated, and dried at 50 °C to obtain the chloroacetamidoquine shown in formula (VI) oxazoline yellow solid, yield 95.6%, melting point 255～258℃. ¹ H NMR (500 MHz, CDCl ₃ ) δ: 3.26-3.33 (m, 1H), 3.54 (dt, J=3.7, 15.4 Hz, 1H), 3.74 (s, 3H), 3.81-3.82 (m, 7H), 3.95-4.05(m,2H),4.28(s,2H),4.64(dd,J=8.2,14.4Hz,1H),5.24(t,J=8.8Hz,1H).6.64(s,1H),6.88 (d, J=8.8Hz, 2H), 7.07 (d, J=8.7Hz, 2H), 7.53 (dd, J=2.3, 9.0Hz, 1H), 7.83 (d, J=9.0Hz, 1H), 8.54 (s, 1H), 8.60 (s, 1H), 8.69 (d, J=2.2 Hz, 1H). IR(KBr,cm ^-1 )ν: 3396, 2998, 2937, 2835, 1694, 1557, 1525, 1510, 1489, 1463, 1349, 1249, 1179, 1036, 840.

Example 12: Preparation of chloroacetamidoquinazoline (VI)

0.27 g (0.55 mmol) of 6-aminoquinazoline (V) prepared by the method of Example 8, 0.04 g (0.55 mmol) of diethylamine, and 10.0 ml of chloroform were successively added to a 50 ml reaction flask, and the conditions were stirred at 10°C. 0.07 g (0.55 mmol) of chloroacetyl chloride and 5.0 mL of chloroform mixed solution were added dropwise, and the dropping was completed, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1 (v/v)), under the condition of 10°C The reaction was carried out for 8 hours, filtered, the filtrate was evaporated to remove the solvent, the concentrate was dissolved in 20 ml of ethanol to obtain a dissolving solution, and 0.26 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) was added to the dissolving solution, and after mixing , evaporate the solvent to obtain a mixture of dry concentrate and silica gel, pack the mixture into a column, then take the petroleum ether/ethyl acetate mixed solution with a volume ratio of 1:5 as the eluent, elution, and TLC tracking detection (development The solvent is ethyl acetate/petroleum ether=1:1 (v/v)), and the eluate containing the compound represented by formula (VI) is collected according to TLC detection (Rf value is 0.5), the collected solution is concentrated and dried at 50°C The chloroacetamidoquinazoline represented by the formula (VI) was obtained as a yellow solid with a yield of 83.4% and a melting point of 255-258°C. ¹ H NMR and IR were the same as in Example 11.

Example 13: Preparation of chloroacetamidoquinazoline (VI)

0.27 g (0.55 mmol) of 6-aminoquinazoline (V) prepared by the method of Example 9, 0.111 g (1.10 mmol) of triethylamine, and 10.0 ml of ethyl acetate were successively added to a 50-ml reaction flask, at 0° C. 0.14 g (1.09 mmol) of chloroacetyl chloride and 5.0 ml of ethyl acetate solution were added dropwise under stirring conditions, and the dropping was completed, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1), and the reaction was carried out at 25°C for 6 20 ml of chloroform was added to dissolve the concentrate to obtain a dissolving solution, 0.30 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) was added to the dissolving solution, and after mixing, evaporated Remove the solvent to obtain a mixture of dry concentrate and silica gel, pack the mixture into a column, and then use the petroleum ether/ethyl acetate mixed solution with a volume ratio of 10:1 as the eluent, elution, and TLC tracking detection (developing solvent is Ethyl acetate/petroleum ether=1:1 (v/v)), collect the eluate containing the compound represented by formula (VI) according to TLC detection (Rf value is 0.5), the collected solution is concentrated, and dried at 50 °C to obtain formula The chloroacetamidoquinazoline shown in (VI) is a yellow solid, the yield is 70.5%, and the melting point is 255-258°C. ¹ H NMR and IR were the same as in Example 11.

Example 14: Preparation of chloroacetamidoquinazoline (VI)

0.27 g (0.55 mmol) of 6-aminoquinazoline (V) prepared by the method of Example 10, 0.067 g (0.55 mmol) of 4-dimethylaminopyridine, and 20.0 ml of toluene were successively added to a 50 ml reaction flask, and 5 The solution of 0.376 g (2.20 mmol) of chloroacetic anhydride and 7.0 ml of toluene was added dropwise under the stirring condition of ℃, and the dropping was completed, heated to 50 ℃, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1), the reaction 3 hours, filtered, the filtrate was evaporated to remove the solvent, the concentrate was dissolved in 20 ml of tetrahydrofuran to obtain a dissolving solution, 0.40 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) was added to the dissolving solution, and after mixing, The solvent was evaporated to obtain a mixture of dry concentrate and silica gel, the mixture was packed into a column, and then the petroleum ether/ethyl acetate mixed solution with a volume ratio of 5:1 was used as the eluent, and the elution was carried out by TLC tracking detection (developing solvent). is ethyl acetate/petroleum ether=1:1 (v/v)), according to TLC detection, the eluate containing the compound represented by formula (VI) (Rf value is 0.5) is collected, the collected solution is concentrated, and dried at 50 °C to obtain The chloroacetamidoquinazoline represented by the formula (VI) is a yellow solid, the yield is 85.3%, and the melting point is 255-258°C. ¹ H NMR and IR were the same as in Example 11.

Example 15: Preparation of chloroacetamidoquinazoline (VI)

0.27 g (0.55 mmol) of 6-aminoquinazoline (V) prepared by the method of Example 7, 0.213 g (1.65 mmol) of quinoline, and 15.0 ml of benzene were successively added to a 50 ml reaction flask, and stirred at -10°C A solution of 0.28 g (2.19 mmol) of chloroacetyl chloride and 5.0 ml of benzene was added dropwise, and the dropping was completed, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1), and the reaction was carried out under -10°C for 12 hours. Filtration, the filtrate was evaporated to remove the solvent, the concentrate was dissolved in 20 ml of tetrahydrofuran to obtain a dissolving solution, 0.40 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) was added to the dissolving solution, and after mixing, the solvent was evaporated , obtain a mixture of dry concentrate and silica gel, pack the mixture into a column, then take the petroleum ether/ethyl acetate mixed solution with a volume ratio of 1:1 as the eluent, elute, and TLC tracking detection (developing solvent is ethyl acetate Ester/petroleum ether=1:1 (v/v)), according to TLC detection, collect the eluate containing the compound represented by formula (VI) (Rf value is 0.5), concentrate the collected solution, and dry at 50 °C to obtain formula (VI) ) as a yellow solid of chloroacetamidoquinazoline, with a yield of 82.1% and a melting point of 255-258°C. ¹ H NMR and IR were the same as in Example 11.

Example 16: Preparation of chloroacetamidoquinazoline (VI)

0.27 g (0.55 mmol) of 6-aminoquinazoline (V) prepared by the method of Example 7, 0.164 g (1.10 mmol) of 4-pyrrolidinopyridine, and 15.0 ml of dichloromethane were successively added to a 50 ml reaction flask , 00.14 g (1.09 mmol) of chloroacetyl chloride and 5.0 ml of dichloromethane solution were added dropwise under stirring conditions at 10 °C, and the dropping was completed, TLC tracking detection (developing solvent was ethyl acetate/petroleum ether=1:1), 10 °C condition The reaction was carried out for 8 hours, filtered, and the filtrate was evaporated to remove the solvent. The concentrate was dissolved in 20 ml of ethanol to obtain a dissolving solution. To the dissolving solution, 0.50 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) was added, and the mixture was uniformly mixed. After that, the solvent was evaporated to obtain a mixture of dry concentrate and silica gel, the mixture was packed into a column, and then the petroleum ether/ethyl acetate mixed solution with a volume ratio of 10:1 was used as the eluent, elution was carried out, and TLC tracking detection ( The developing solvent is ethyl acetate/petroleum ether=1:1 (v/v)), and the eluate containing the compound represented by formula (VI) is collected according to TLC detection (Rf value is 0.5), the collected solution is concentrated, 50 ℃ After drying, the yellow solid of chloroacetamidoquinazoline represented by formula (VI) was obtained, the yield was 90.2%, and the melting point was 255-258°C. ¹ H NMR and IR were the same as in Example 11.

基喹唑啉(Ⅰ)的制备Example 17: Morpholinylacetamidobenzo[d]azepine

Preparation of quinazoline (Ⅰ)

3.25 g (5.73 mmol) of chloroacetamidoquinazoline (VI) and 0.598 g (6.86 mmol) of morpholine, 3.626 g (45.84 mmol) of pyridine, and 32.5 ml of methanol prepared by the method of Example 11 were successively added to a 50-ml reaction flask. , heated to 40°C, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1 (v/v)), stirred and reacted for 10 hours, closed the reaction, evaporated the solvent from the reaction solution, and obtained in the concentrate Add 10 ml of ethyl acetate to dissolve it to obtain a dissolving solution, add 1.5 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the dissolving solution, and after mixing, evaporate the solvent to obtain a dry concentrate and A mixture of silica gel, the mixture is packed into a column, and then the eluent is eluted with a petroleum ether/ethyl acetate mixed solution with a volume ratio of 1:10, followed by TLC detection (developing solvent is ethyl acetate/petroleum ether=1: 1(v/v)), collect the eluate containing the compound shown in formula (I) according to TLC detection (Rf value is 0.5), the collected solution is concentrated, and dried at 50 °C to obtain the off-white solid product shown in formula (I). , the yield is 65.4%, the melting point is 122～125℃. ¹ H NMR (500MHz, CDCl ₃ )δ: 2.65-2.72(m, 4H), 3.19(s, 2H), 3.30(m, 1H), 3.54(d, J=15.3Hz, 1H), 3.74(s, 3H), 3.81-3.84(m, 11H), 3.99-4.01(m, 2H), 4.64(dd, J=8.2.14.2Hz, 1H), 5.27(t, J=8.6Hz, 1H), 6.67(s ,1H),6.87(d,J＝8.6Hz,2H),7.07(d,J＝8.6Hz,2H),7.40(dd,J＝2.0,8.9Hz,1H),7.81(d,J＝8.9Hz , 1H), 8.58(s, 1H), 8.84(s, 1H), 9.29(s, 1H). HRMS-ESI m/z: 618.2477[M+H] ⁺ . IR(KBr,cm ^-1 )ν: 2933, 2833, 1692, 1609, 1523, 1568, 1523, 1488, 1461, 1348, 1248, 1116, 1035, 838.

基喹唑啉(Ⅰ)的制备Example 18: Morpholinylacetamidobenzo[d]azepine

Preparation of quinazoline (Ⅰ)

Add 3.25 g (5.73 mmol) of chloroacetamidoquinazoline (VI) and 0.398 g (4.57 mmol) of morpholine, 2.95 g (22.84 mmol) of quinoline, and 80 ml of toluene to 100 ml of three-hole The flask was heated to 100° C., followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1 (v/v)), stirred and reacted for 2 hours, closed the reaction, evaporated the solvent from the reaction solution, and obtained the concentrate Add 20 ml of ethanol to dissolve it to obtain a dissolving solution, add 2.5 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the dissolving solution, and after mixing, evaporate the solvent to obtain a dry concentrate and silica gel The mixture was packed into a column, and then the eluent was eluted with a petroleum ether/ethyl acetate mixed solution with a volume ratio of 1:5, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1 (v/v)), collect the eluate containing the compound represented by formula (I) according to TLC detection (Rf value is 0.5), concentrate the collected solution, and dry at 50 °C to obtain the off-white solid product represented by formula (I), The yield is 61.6%, and the melting point is 122-125°C. ¹ H NMR and IR were the same as in Example 17.

基喹唑啉(Ⅰ)的制备Example 19: Morpholinylacetamidobenzo[d]azepine

Preparation of quinazoline (Ⅰ)

3.25 g (5.73 mmol) of chloroacetamidoquinazoline (VI) prepared by the method of Example 13, 0.499 g (5.73 mmol) of morpholine, 0.58 g (5.73 mmol) of triethylamine, and 80 ml of ethanol were added to 100 ml of ethanol in turn. In a three-necked flask, heated to 60°C, followed by TLC detection (developing solvent is ethyl acetate/petroleum ether=1:1 (v/v)), stirred and reacted for 8 hours, closed the reaction, evaporated the solvent from the reaction solution, and obtained concentrated Add 20 ml of chloroform to dissolve it to obtain a dissolving solution, add 2.5 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the dissolving solution, and after mixing, evaporate the solvent to obtain a dry concentrate with A mixture of silica gel, the mixture was packed into a column, and then a mixed solution of petroleum ether/ethyl acetate with a volume ratio of 10:1 was used as the eluent, eluted, and TLC tracking detection (developing solvent was ethyl acetate/petroleum ether=1: 1(v/v)), collect the eluate containing the compound shown in formula (I) according to TLC detection (Rf value is 0.5), the collected solution is concentrated, and dried at 50 °C to obtain the off-white solid product shown in formula (I). , the yield is 57.7%, the melting point is 122～125℃. ¹ H NMR and IR were the same as in Example 17.

基喹唑啉(Ⅰ)的制备Example 20: Morpholinylacetamidobenzo[d]azepine

Preparation of quinazoline (Ⅰ)

3.25 g (5.73 mmol) chloroacetamidoquinazoline (VI) prepared by the method of Example 14, 1.997 g (22.92 mmol) morpholine, 1.40 g (11.46 mmol) 4-dimethylaminopyridine, 60 mL isopropyl The alcohol was added to a 100-mL three-necked flask, stirred at room temperature at 25°C, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1 (v/v)), reacted for 36 hours, closed the reaction, and evaporated the solvent from the reaction solution. , add 20 ml of tetrahydrofuran to the obtained concentrate to dissolve it to obtain a dissolving solution, add 3.0 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the dissolving solution, and after mixing, evaporate the solvent to obtain a dry The mixture of the concentrate and silica gel, the mixture is packed into a column, and then take the petroleum ether/ethyl acetate mixed solution with a volume ratio of 5:1 as the eluent, elution, TLC tracking detection (developing solvent is ethyl acetate/petroleum ether=1:1 (v/v)), according to TLC detection, collect the eluate containing the compound represented by formula (I) (Rf value is 0.5), concentrate the collected solution, and dry at 50°C to obtain the compound represented by formula (I) The off-white solid product, the yield was 71.3%, and the melting point was 122-125°C. ¹ H NMR and IR were the same as in Example 17.

基喹唑啉(Ⅰ)的制备Example 21: Morpholinylacetamidobenzo[d]azepine

Preparation of quinazoline (Ⅰ)

3.25 g (5.73 mmol) chloroacetamidoquinazoline (VI) and 0.449 g (5.15 mmol) morpholine prepared by the method of Example 15, 1.04 g (8.58 mmol) N,N-xylidine, 33 mL N , N-dimethylformamide was added to a 50 ml reaction flask, heated to 120°C, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1 (v/v)), and the reaction was stirred for 0.5 hours. The reaction was closed, the solvent was evaporated from the reaction solution, 20 ml of tetrahydrofuran was added to the obtained concentrate to dissolve it to obtain a dissolving solution, 4.0 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) was added to the dissolving solution, and the mixture was uniformly mixed. Then, the solvent was evaporated to obtain a mixture of dry concentrate and silica gel, the mixture was packed into a column, and then the petroleum ether/ethyl acetate mixed solution with a volume ratio of 1:1 was used as the eluent, elution was carried out, and TLC tracking detection ( The developing solvent is ethyl acetate/petroleum ether=1:1 (v/v)), and the eluate containing the compound represented by formula (I) is collected according to TLC detection (Rf value is 0.5), the collected solution is concentrated, 50 ℃ After drying, the off-white solid product represented by formula (I) was obtained, the yield was 55.2%, and the melting point was 122-125°C. ¹ H NMR and IR were the same as in Example 17.

基喹唑啉(Ⅰ)的制备Example 22: Morpholinylacetamidobenzo[d]azepine

Preparation of quinazoline (Ⅰ)

3.25 g (5.73 mmol) of chloroacetamidoquinazoline (VI) prepared by the method of Example 16, 3.994 g (45.84 mmol) of morpholine, 3.626 g (45.84 mmol) of pyridine, and 195 ml of propanol were added to the reaction of 500 ml. In the bottle, heated to 40°C, followed by TLC detection (developing solvent is ethyl acetate/petroleum ether=1:1 (v/v)), stirred and reacted for 10 hours, closed the reaction, evaporated the solvent from the reaction solution, and obtained the concentrate Add 20 ml of ethyl acetate to dissolve it to obtain a dissolving solution, add 6.0 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the dissolving solution, and after mixing, evaporate the solvent to obtain a dry concentrate The mixture with silica gel, pack the mixture into a column, and then use the petroleum ether/ethyl acetate mixed solution with a volume ratio of 1:1 as the eluent, elution, and TLC tracking detection (developing solvent is ethyl acetate/petroleum ether=1 : 1 (v/v)), according to TLC detection, collect the eluate containing the compound shown in formula (I) (Rf value is 0.5), the collected solution is concentrated, and dried at 50 °C to obtain the off-white solid shown in formula (I). Product, yield 75.7%, melting point 122～125℃. ¹ H NMR and IR were the same as in Example 17.

Example 23: In vitro testing of anticancer activity

(1) The prepared compound (I) was tested for biological activity of human cervical cancer cell line Siha.

Test method: Tetrazolium salt reduction method (MTT method).

Cell line: human cervical cancer cell line Siha. The above tumor cell lines were purchased from the Cell Bank of Shanghai Academy of Biological Sciences, Chinese Academy of Sciences.

The experimental steps are as follows:

(a) Sample preparation: For soluble samples, dissolve each 1 mg with 40 μL DMSO, take 2 μL and dilute with 1000 μL medium to make the concentration 100 μg/mL, and then serially dilute with culture medium to the use concentration.

(b) Culture of cells

① Preparation of medium: Each 1000 mL of DMEM medium (Gibco) contains 800,000 units of penicillin, 1.0 g of streptomycin, and 10% inactivated fetal bovine serum.

② Cell culture: The tumor cells were inoculated into the medium, cultured in a 37°C, 5% CO ₂ incubator, and passaged for 3-5 days.

③ Determination of the inhibitory effect of samples on tumor cell growth

The second passage cells were digested with EDTA-trypsin digestion solution, diluted with medium to 1×10 ⁶ /mL, added to a 96-well cell culture plate, 100 μL per well, and placed in a 37°C, 5% CO ₂ incubator nourish. 24h after inoculation, 100μg/mL, 10μg/mL and 1μg/mL samples diluted with culture medium were added, 100μL per well, 3 wells for each concentration, and cultured in a 37°C, 5% _CO2 incubator for 72h. Add 5 mg/mL MTT to the cell culture wells, 10 μL per well, incubate at 37°C for 3 h, add DMSO, 150 μL per well, shake with a shaker to completely dissolve the formazan, and use a microplate reader for colorimetry at a wavelength of 570 nm . Taking the cells cultured in the medium containing the same concentration of DMSO without the sample under the same conditions as a control, the IC ₅₀ of the sample on the growth of tumor cells was calculated. The test results are shown in Table 1:

Table 1. Inhibitory effect of compound (I) on the growth of cancer cell line Siha

(2) according to embodiment 11, chloroacetyl chloride is respectively replaced with 3-methoxybenzoyl chloride or cinnamoyl chloride, other operations are the same as embodiment 11, quinazoline compounds (b) and (c) are synthesized respectively, The structure looks like this:

According to the above method, the prepared quinazoline compounds (b) and (c) were tested for biological activity of human cervical cancer cell line Siha. The inhibitory effect of cell line Siha was not obvious, and the anticancer activity of compounds (b) and (c) on human cervical cancer cell line Siha was far inferior to that of compound (I). The specific results are shown in Table 2:

Table 2. Inhibitory effect of compounds (b) and (c) on the growth of cancer cell line Siha

The above-mentioned anticancer activity in vitro test experiments show that the other two compounds (b) and (c) with similar structures have no obvious inhibitory effect on the growth of human cervical cancer cell line Siha. Compound (I) has a significant inhibitory effect on the growth of human cervical cancer cell line Siha, which is significantly better than that of compounds (b) and (c).

(3) According to Example 17, morpholine was replaced with 3,4-xylidine, 3,4-dimethoxyaniline or di-n-propylamine respectively, and other operations were the same as in Example 17, and quinazolines were synthesized respectively. Compounds (d), (e) and (f), the structures are shown below:

According to the above method, the prepared quinazoline compounds (d), (e) and (f) were tested for biological activity of human cervical cancer cell line Siha, and the results showed that the quinazoline compounds (d), (e) and (f) The anticancer activity against human cervical cancer cell line Siha is far inferior to compound (I). The specific results are shown in Table 3:

Table 3. Inhibitory effect of compounds (d), (e) and (f) on the growth of cancer cell line Siha

Claims (2)

1. Morpholinyl acetamidobenzo [ d ] as shown in formula (I)]Aza derivatives

The application of the quinazoline compound in preparing the medicine for preventing or treating the cervical cancer of the human;

2. the use according to claim 1, wherein the medicament is a medicament having inhibitory activity against the human cervical cancer cell line, Siha.