CN108014115B - Application of pivaloylaminomethoxyphenyl benzo [ d ] aza-quinazoline compound in preparation of lung cancer treatment drug - 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肺癌药物中的应用，其对抑制人肺癌细胞株A‑549活性具有显著效果。The invention discloses a pivalamidomethoxyphenylbenzo[d]azepine

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 lung cancer, has a significant effect on inhibiting the activity of human lung cancer cell line A-549.

Description

Pivalamidomethoxyphenyl benzo[d]aza*ylquinazoline compounds are being prepared Application in the preparation of drugs for the treatment of lung cancer

(1) Technical field

基喹唑啉类化合物在制备预防或治疗人肺癌药物中的应用。The present invention relates to the application of a kind of quinazoline compounds, particularly a kind of pivalamido methoxyphenyl benzo[d]azepine

Application of quinazoline compounds in the preparation of medicaments for preventing or treating human lung 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肺癌细胞株A-549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 - pivalamido methoxy phenyl benzo [d] aza

The application of quinazoline compounds, the compounds have a good inhibitory effect on the human lung cancer cell line A-549 under a certain dose; and the preparation method of the compounds is simple, easy to operate, easy to obtain raw materials, and relatively low production cost. low, 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肺癌药物中的应用：The present invention provides a pivalamidomethoxyphenylbenzo[d]azepine represented by formula (I)

Application of quinazoline compounds in the preparation of drugs for preventing or treating tumors, especially in the preparation of drugs for preventing or treating human lung cancer:

Preferably, the drug is a drug with the activity of inhibiting human lung cancer cell line A-549.

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 pivalamidomethoxyphenylbenzo[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 0.5-12 h), the reaction solution is filtered, the filtrate is concentrated under reduced pressure and the concentrate 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 pivaloyl chloride or pivalic 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 post-treated to obtain the compound represented by formula (I); the basic Catalyst F is one of the following: pyridine, diethylamine, triethylamine, quinoline, N,N-xylidine, 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.

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 pivaloyl chloride or pivalic anhydride and the amount of the basic catalyst F charged 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 Add the organic solvent G solution of pivaloyl chloride or pivalic anhydride dropwise to the indicated compound and the basic catalyst F, and after the dropping is completed, react at -10～50℃ for 3～12 hours, and the obtained reaction solution is subjected to post-treatment to obtain the formula (I) shown in the formula (I). Compound; the volume dosage of the organic solvent for dissolving pivaloyl chloride or pivalic 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 pivaloyl chloride or pivalic anhydride.

Further, the method for post-treatment of the reaction solution in step (3) of the present invention is as follows: after the reaction is complete, 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 dissolving 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 used as the eluent, and the effluent containing the target component is collected (preferably, ethyl acetate/petroleum ether=1:1 (v/v) is used as the developing agent to track and detect, and the target component is collected) , concentrated under reduced pressure, and dried (preferably at 50° C.) to obtain the compound represented by formula (I); the organic solvent H is one of the following: ethanol, chloroform, tetrahydrofuran or ethyl acetate. The amount of the organic solvent H can be used to dissolve the residue.

The organic solvents A, C, D, G and H 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, the reducing agent E and the catalyst F are all Catalysts are named in order to distinguish the different catalysts used in different steps, and the letters themselves have no meaning.

基喹唑啉类化合物(Ⅰ)在制备预防或治疗人肺癌的药物中的应用，该化合物对人肺癌细胞株A-549具有显著的抑制活性。The beneficial effects of the present invention are mainly reflected in: providing a novel pivalamidomethoxyphenylbenzo[d]azepine

The application of the quinazoline compound (I) in the preparation of a medicament for preventing or treating human lung cancer, the compound has significant inhibitory activity on the human lung cancer cell line A-549.

(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.

基喹唑啉(Ⅳ)的制备Example 1: Nitrobenzo[d]azepine

Preparation of quinazoline (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.

基喹唑啉(Ⅳ)的制备Example 2: Nitrobenzo[d]azepine

Preparation of quinazoline (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.

基喹唑啉(Ⅳ)的制备Example 3: Nitrobenzo[d]azepine

Preparation of quinazoline (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.

基喹唑啉(Ⅳ)的制备Example 4: Nitrobenzo[d]azepine

Preparation of quinazoline (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.

基喹唑啉(Ⅳ)的制备Example 5: Nitrobenzo[d]azepine

Preparation of quinazoline (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.

基喹唑啉(Ⅳ)的制备Example 6: Nitrobenzo[d]azepine

Preparation of quinazoline (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.

基喹唑啉(Ⅴ)的制备Example 7: Aminobenzo[d]azepine

Preparation of quinazoline (V)

基喹唑啉(Ⅳ)，0.40克(6.34mmol)甲酸铵，0.04克5％Pd/C，4.0毫升氯仿加入到反应瓶中，室温25℃搅拌，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1(v/v))，反应12小时，过滤，滤液浓缩，25℃真空干燥得到淡黄色固体产物氨基苯并[d]氮杂

基喹唑啉(Ⅴ)，收率98.2％，熔点122～126℃。¹H NMR(500MHz,CDCl₃)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。0.40 g (0.77 mmol) of nitrobenzo[d]azepine prepared by the method of Example 1

Quinazoline (IV), 0.40 g (6.34 mmol) of ammonium formate, 0.04 g of 5% Pd/C, and 4.0 ml of chloroform were added to the reaction flask, stirred at room temperature at 25°C, and detected by TLC (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 light yellow solid product aminobenzo[d]azepine

quinazoline (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.

基喹唑啉(Ⅴ)的制备Example 8: Aminobenzo[d]azepine

Preparation of quinazoline (V)

基喹唑啉(Ⅳ)，1.20克(19.18mmol)80wt％水合肼，0.20克5％Pd/C，20.0毫升甲苯加入到50毫升的反应瓶中，加热至100℃，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1(v/v))，搅拌反应0.5小时，冷却过滤，滤液浓缩，25℃真空干燥得到淡黄色固体产物氨基苯并[d]氮杂

基喹唑啉(Ⅴ)，收率100.0％，熔点122～126℃。¹H NMR和IR同实施例7。0.40 g (0.77 mmol) of nitrobenzo[d]azepine prepared by the method of Example 2

Quinazoline (IV), 1.20 g (19.18 mmol) of 80wt% hydrazine hydrate, 0.20 g of 5% Pd/C, 20.0 ml of toluene were added to a 50 ml reaction flask, heated to 100 ° C, TLC tracking detection (developing agent) 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 to obtain aminobenzo[d]azepine as a light yellow solid product

quinazoline (V), yield 100.0%, melting point 122～126℃. ¹ H NMR and IR were the same as in Example 7.

基喹唑啉(Ⅴ)的制备Example 9: Aminobenzo[d]azepine

Preparation of quinazoline (V)

基喹唑啉(Ⅳ)，0.08克浓盐酸(质量浓度36～38％)，0.40克铁粉，20.0毫升甲醇加入到50毫升的反应瓶中，加热至40℃，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1(v/v))，搅拌反应8小时，冷却过滤，滤液浓缩，25℃真空干燥得到淡黄色固体产物氨基苯并[d]氮杂

基喹唑啉(Ⅴ)，收率94.1％，熔点122～126℃。¹H NMR和IR同实施例7。0.40 g (0.77 mmol) of nitrobenzo[d]azepine prepared by the method of Example 3

Quinazoline (IV), 0.08 g concentrated hydrochloric acid (mass concentration 36～38%), 0.40 g iron powder, 20.0 ml methanol were added to 50 ml reaction flask, heated to 40°C, followed by TLC detection (developing agent 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 aminobenzo[d]azepine as a light yellow solid product

quinazoline (V), yield 94.1%, melting point 122～126℃. ¹ H NMR and IR were the same as in Example 7.

基喹唑啉(Ⅴ)的制备Example 10: Aminobenzo[d]azepine

Preparation of quinazoline (V)

基喹唑啉(Ⅳ)，0.40克醋酸，1.20克铁粉，20.0毫升异丙醇加入到50毫升的反应瓶中，加热至80℃，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1(v/v))，搅拌反应3小时，冷却过滤，滤液浓缩，25℃真空干燥得到淡黄色固体产物氨基苯并[d]氮杂

基喹唑啉(Ⅴ)，收率97.5％，熔点122～126℃。¹H NMR和IR同实施例7。0.40 g (0.77 mmol) of nitrobenzo[d]azepine prepared by the method of Example 4

Quinazoline (IV), 0.40 g of acetic acid, 1.20 g of iron powder, 20.0 ml of isopropanol were added to a 50 ml reaction flask, heated to 80° C., followed by TLC detection (developing solvent was ethyl acetate/petroleum ether= 1:1 (v/v)), stirred and reacted for 3 hours, cooled and filtered, the filtrate was concentrated, and vacuum-dried at 25°C to obtain aminobenzo[d]azepine as a light yellow solid product

quinazoline (V), yield 97.5%, melting point 122～126℃. ¹ H NMR and IR were the same as in Example 7.

基喹唑啉(Ⅰ)的制备Example 11: Pivalamidomethoxyphenylbenzo[d]azepine

Preparation of quinazoline (Ⅰ)

基喹唑啉(Ⅴ)，0.13克(1.64mmol)吡啶，3毫升四氢呋喃加入到反应瓶中，-10℃搅拌条件下滴加0.531克(4.40mmol)特戊酰氯，滴毕，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1)，-10℃条件下反应12小时，过滤，滤液蒸除溶剂，浓缩物加入10毫升乙酸乙酯将其溶解，获得溶解液，向溶解液中加入0.60克柱层析硅胶(300～400目柱层析硅胶)，混匀后，蒸除溶剂，得干燥的浓缩物与硅胶的混合物，将混合物装柱，然后以体积比为1：10的石油醚/乙酸乙酯混合溶液为洗脱剂，洗脱，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1(v/v))，根据TLC检测收集含式(Ⅰ)所示的化合物的洗脱液(Rf值为0.5)，收集液浓缩，50℃干燥得到式(Ⅰ)所示的特戊酰氨基甲氧苯基苯并[d]氮杂

基喹唑啉灰白色固体，收率64.2％，熔点131～133℃。¹H NMR(500MHz,[D₆]DMSO)δ:1.29(s,9H),3.22-3.28(m,1H),3.38-3.42(m,1H),3.68(s,3H),3.69(s,3H),3.73(s,3H),3.78-3.84(m,1H),3.87-3.96(m,2H),4.49(dd,J＝8.2,14.7,1H),5.26(t,J＝8.6Hz,1H),6.85(s,1H),6.89(d,J＝8.8Hz,2H),7.08(d,J＝8.7Hz,2H),7.69(d,J＝9.0,1H),7.88(dd,J＝2.2,9.2Hz,1H),8.44(s,1H),8.66(s,1H),9.55(s,1H)。HRMS-ESI m/z:575.2417[M+H]⁺。IR(KBr,cm^-1)ν:2966,2921,2868,1665,1557,1522,1510,1490,1163,1349,1248,1036,847。0.27 g (0.55 mmol) of aminobenzo[d]azepine prepared by the method of Example 7

Quinazoline (V), 0.13 g (1.64 mmol) of pyridine, and 3 ml of tetrahydrofuran were added to the reaction flask, and 0.531 g (4.40 mmol) of pivaloyl chloride was added dropwise with stirring at -10°C. The developing solvent was ethyl acetate/petroleum ether=1:1), reacted at -10°C for 12 hours, filtered, the filtrate was evaporated to remove the solvent, and the concentrate was dissolved in 10 ml of ethyl acetate to obtain a dissolving solution. 0.60 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) was added to the mixture, 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 the volume ratio was 1:10 The petroleum ether/ethyl acetate mixed solution is the eluent, elution, TLC tracking detection (developing solvent is ethyl acetate/petroleum ether=1:1 (v/v)), according to TLC detection collection containing formula (I) The eluate of the compound shown (Rf value is 0.5), the collected solution is concentrated, and dried at 50 °C to obtain the pivalamidomethoxyphenylbenzo[d]azepine represented by the formula (I).

The quinazoline was an off-white solid, the yield was 64.2%, and the melting point was 131-133°C. ¹ H NMR (500MHz, [D ₆ ]DMSO)δ: 1.29(s, 9H), 3.22-3.28(m, 1H), 3.38-3.42(m, 1H), 3.68(s, 3H), 3.69(s, 3H), 3.73(s, 3H), 3.78-3.84(m, 1H), 3.87-3.96(m, 2H), 4.49(dd, J=8.2, 14.7, 1H), 5.26(t, J=8.6Hz, 1H),6.85(s,1H),6.89(d,J=8.8Hz,2H),7.08(d,J=8.7Hz,2H),7.69(d,J=9.0,1H),7.88(dd,J = 2.2, 9.2 Hz, 1H), 8.44 (s, 1H), 8.66 (s, 1H), 9.55 (s, 1H). HRMS-ESI m/z: 575.2417[M+H] ⁺ . IR(KBr,cm ^-1 )ν: 2966, 2921, 2868, 1665, 1557, 1522, 1510, 1490, 1163, 1349, 1248, 1036, 847.

基喹唑啉(Ⅰ)的制备Example 12: Pivalamidomethoxyphenylbenzo[d]azepine

Preparation of quinazoline (Ⅰ)

基喹唑啉(Ⅴ)，0.04克(0.55mmol)二乙胺，10.0毫升氯仿加入到50毫升的反应瓶中，10℃搅拌条件下滴加0.066克(0.55mmol)特戊酰氯和5.0毫升氯仿混合溶液，滴毕，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1(v/v))，10℃条件下反应8小时，过滤，滤液蒸除溶剂，浓缩物加入20毫升乙醇将其溶解，获得溶解液，向溶解液中加入0.26克柱层析硅胶(300～400目柱层析硅胶)，混匀后，蒸除溶剂，得干燥的浓缩物与硅胶的混合物，将混合物装柱，然后以体积比为1：5的石油醚/乙酸乙酯混合溶液为洗脱剂，洗脱，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1(v/v))，根据TLC检测收集含式(Ⅰ)所示的化合物的洗脱液(Rf值为0.5)，收集液浓缩，50℃干燥得到式(Ⅰ)所示的特戊酰氨基甲氧苯基苯并[d]氮杂

基喹唑啉灰白色固体，收率74.7％，熔点131～133℃。¹H NMR和IR同实施例11。0.27 g (0.55 mmol) aminobenzo[d]azepine prepared by the method of Example 8

Quinazoline (V), 0.04 g (0.55 mmol) diethylamine, and 10.0 mL chloroform were added to a 50 mL reaction flask, and 0.066 g (0.55 mmol) pivaloyl chloride and 5.0 mL chloroform were added dropwise with stirring at 10°C The mixed solution was dripped, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1 (v/v)), reacted at 10°C for 8 hours, filtered, the filtrate was evaporated to remove the solvent, and 20 ml of the concentrate was added Dissolve it in ethanol to obtain a dissolving solution, add 0.26 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 mixture of dry concentrate and silica gel. The mixture was packed into a column, and then a mixed solution of petroleum ether/ethyl acetate with a volume ratio of 1:5 was used as the eluent, eluted, and 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 (I) (Rf value is 0.5), the collected solution is concentrated, and dried at 50 °C to obtain the pivalamid methoxyphenyl shown in formula (I). Benzo[d]azepine

The quinazoline was an off-white solid, the yield was 74.7%, and the melting point was 131-133°C. ¹ H NMR and IR were the same as in Example 11.

基喹唑啉(Ⅰ)的制备Example 13: Pivalamidomethoxyphenylbenzo[d]azepine

Preparation of quinazoline (Ⅰ)

基喹唑啉(Ⅴ)，0.111克(1.10mmol)三乙胺，10.0毫升乙酸乙酯加入到50毫升的反应瓶中，0℃搅拌条件下滴加0.133克(1.10mmol)特戊酰氯和5.0毫升乙酸乙酯溶液，滴毕，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1)，25℃条件下反应6小时，过滤，滤液蒸除溶剂，浓缩物加入20毫升氯仿将其溶解，获得溶解液，向溶解液中加入0.30克柱层析硅胶(300～400目柱层析硅胶)，混匀后，蒸除溶剂，得干燥的浓缩物与硅胶的混合物，将混合物装柱，然后以体积比为10：1的石油醚/乙酸乙酯混合溶液为洗脱剂，洗脱，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1(v/v))，根据TLC检测收集含式(Ⅰ)所示的化合物的洗脱液(Rf值为0.5)，收集液浓缩，50℃干燥得到式(Ⅰ)所示的特戊酰氨基甲氧苯基苯并[d]氮杂

基喹唑啉灰白色固体，收率50.5％，熔点131～133℃。¹H NMR和IR同实施例11。0.27 g (0.55 mmol) aminobenzo[d]azepine prepared by the method of Example 9

Quinazoline (V), 0.111 g (1.10 mmol) triethylamine, 10.0 ml ethyl acetate were added to a 50 ml reaction flask, and 0.133 g (1.10 mmol) pivaloyl chloride and 5.0 ml of ethyl acetate solution, dripped, followed by TLC (developing solvent: ethyl acetate/petroleum ether=1:1), reacted at 25°C for 6 hours, filtered, the filtrate was evaporated to remove the solvent, and 20 ml of chloroform was added to the concentrate to make the solution. It is dissolved to obtain a dissolving solution, 0.30 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) is added to the dissolving solution, and after mixing, the solvent is evaporated to obtain a mixture of dry concentrate and silica gel, and the mixture is packed. 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)) , 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 pivalamidomethoxyphenyl benzoyl compound shown in formula (I). [d]Aza

The quinazoline was an off-white solid, the yield was 50.5%, and the melting point was 131-133°C. ¹ H NMR and IR were the same as in Example 11.

基喹唑啉(Ⅰ)的制备Example 14: Pivalamidomethoxyphenylbenzo[d]azepine

Preparation of quinazoline (Ⅰ)

基喹唑啉(Ⅴ)，0.067克(0.55mmol)4-二甲氨基吡啶，20.0毫升甲苯加入到50毫升的反应瓶中，5℃搅拌条件下滴加0.410克(2.20mmol)特戊酸酐和7.0毫升甲苯的溶液，滴毕，加热至50℃，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1)，反应3小时，过滤，滤液蒸除溶剂，浓缩物加入20毫升四氢呋喃将其溶解，获得溶解液，向溶解液中加入0.40克柱层析硅胶(300～400目柱层析硅胶)，混匀后，蒸除溶剂，得干燥的浓缩物与硅胶的混合物，将混合物装柱，然后以体积比为5：1的石油醚/乙酸乙酯混合溶液为洗脱剂，洗脱，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1(v/v))，根据TLC检测收集含式(Ⅰ)所示的化合物的洗脱液(Rf值为0.5)，收集液浓缩，50℃干燥得到式(Ⅰ)所示的特戊酰氨基甲氧苯基苯并[d]氮杂

基喹唑啉灰白色固体，收率57.4％熔点131～133℃。¹H NMR和IR同实施例11。0.27 g (0.55 mmol) of aminobenzo[d]azepine prepared by the method of Example 10

Quinazoline (V), 0.067 g (0.55 mmol) 4-dimethylaminopyridine, 20.0 ml of toluene were added to a 50 ml reaction flask, 0.410 g (2.20 mmol) of pivalic anhydride and 0.410 g (2.20 mmol) of pivalic anhydride and The solution of 7.0 ml of toluene, after dripping, was heated to 50°C, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1), reacted for 3 hours, filtered, the filtrate was evaporated to remove the solvent, and 20 ml of tetrahydrofuran was added to the concentrate Dissolve it to obtain a dissolving solution, add 0.40 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 mixture of dry concentrate and silica gel. Pack the column, then use the petroleum ether/ethyl acetate mixed solution with a volume ratio of 5:1 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 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 pivalamidomethoxyphenylbenzene shown in formula (I). And[d]aza

Quinazoline off-white solid, yield 57.4%, melting point 131-133°C. ¹ H NMR and IR were the same as in Example 11.

基喹唑啉(Ⅰ)的制备Example 15: Pivalamidomethoxyphenylbenzo[d]azepine

Preparation of quinazoline (Ⅰ)

基喹唑啉(Ⅴ)，0.213克(1.65mmol)喹啉，15.0毫升苯加入到50毫升的反应瓶中，-10℃搅拌条件下滴加0.265克(2.20mmol)特戊酰氯和5.0毫升苯的溶液，滴毕，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1)，-10℃条件下反应12小时，过滤，滤液蒸除溶剂，浓缩物加入20毫升四氢呋喃将其溶解，获得溶解液，向溶解液中加入0.40克柱层析硅胶(300～400目柱层析硅胶)，混匀后，蒸除溶剂，得干燥的浓缩物与硅胶的混合物，将混合物装柱，然后以体积比为1：1的石油醚/乙酸乙酯混合溶液为洗脱剂，洗脱，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1(v/v))，根据TLC检测收集含式(Ⅰ)所示的化合物的洗脱液(Rf值为0.5)，收集液浓缩，50℃干燥得到式(Ⅰ)所示的特戊酰氨基甲氧苯基苯并[d]氮杂

基喹唑啉灰白色固体，收率70.9％，熔点131～133℃。¹H NMR和IR同实施例11。0.27 g (0.55 mmol) of aminobenzo[d]azepine prepared by the method of Example 7

Quinazoline (V), 0.213 g (1.65 mmol) of quinoline, and 15.0 mL of benzene were added to a 50-mL reaction flask, and 0.265 g (2.20 mmol) of pivaloyl chloride and 5.0 mL of benzene were added dropwise with stirring at -10°C. The solution was dripped, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1), reacted at -10°C for 12 hours, filtered, the filtrate was evaporated to remove the solvent, and 20 ml of tetrahydrofuran was added to the concentrate to dissolve it. , to obtain a dissolving solution, add 0.40 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the dissolving solution, after mixing, evaporate the solvent to obtain a mixture of dry concentrate and silica gel, pack the mixture into a column, Then use 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 The eluate containing the compound represented by the formula (I) was collected by TLC detection (Rf value was 0.5), the collected liquid was concentrated, and dried at 50°C to obtain the pivalamidomethoxyphenylbenzo[d] represented by the formula (I). ] aza

The quinazoline was an off-white solid, the yield was 70.9%, and the melting point was 131-133°C. ¹ H NMR and IR were the same as in Example 11.

基喹唑啉(Ⅰ)的制备Example 16: Pivalamidomethoxyphenylbenzo[d]azepine

Preparation of quinazoline (Ⅰ)

基喹唑啉(Ⅴ)，0.164克(1.10mmol)4-吡咯烷基吡啶，15.0毫升二氯甲烷加入到50毫升的反应瓶中，10℃搅拌条件下滴加0.133克(1.10mmol)特戊酰氯和5.0毫升二氯甲烷溶液，滴毕，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1)，10℃条件下反应8小时，过滤，滤液蒸除溶剂，浓缩物加入20毫升乙醇将其溶解，获得溶解液，向溶解液中加入0.50克柱层析硅胶(300～400目柱层析硅胶)，混匀后，蒸除溶剂，得干燥的浓缩物与硅胶的混合物，将混合物装柱，然后以体积比为10：1的石油醚/乙酸乙酯混合溶液为洗脱剂，洗脱，TLC跟踪检测(展开剂为乙酸乙酯/石油醚＝1：1(v/v))，根据TLC检测收集含式(Ⅰ)所示的化合物的洗脱液(Rf值为0.5)，收集液浓缩，50℃干燥得到式(Ⅰ)所示的特戊酰氨基甲氧苯基苯并[d]氮杂

基喹唑啉灰白色固体，收率68.6％，熔点131～133℃。¹H NMR和IR同实施例11。0.27 g (0.55 mmol) of aminobenzo[d]azepine prepared by the method of Example 7

Quinazoline (V), 0.164 g (1.10 mmol) 4-pyrrolidinopyridine, 15.0 ml dichloromethane were added to a 50 ml reaction flask, and 0.133 g (1.10 mmol) pivale was added dropwise with stirring at 10°C Acyl chloride and 5.0 ml of dichloromethane solution were dripped, followed by TLC detection (developing solvent was ethyl acetate/petroleum ether=1:1), reacted at 10°C for 8 hours, filtered, the filtrate was evaporated to remove the solvent, and the concentrate was added with 20 It was dissolved in milliliter ethanol to obtain a dissolving solution, 0.50 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, Pack the mixture into a column, and then take the petroleum ether/ethyl acetate mixed solution with a volume ratio of 10:1 as the eluent, elute, and TLC tracking detection (developing solvent is 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 pivalamid methoxybenzene represented by formula (I) benzo[d]azepine

The quinazoline was an off-white solid, the yield was 68.6%, and the melting point was 131-133°C. ¹ H NMR and IR were the same as in Example 11.

Example 17: In vitro testing of anticancer activity

(1) The prepared compounds (I) and (IV) were tested for biological activity of human lung cancer cell line A-549.

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

Cell line: human lung cancer cell line A-549. 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 effects of compounds (I) and (IV) on the growth of cancer cell line A-549

(2) According to Example 11, pivaloyl chloride was replaced with 4-iodobenzoyl chloride, 3-methoxybenzoyl chloride or cinnamoyl chloride respectively, and other operations were the same as in Example 11, and quinazoline compounds were synthesized respectively ( a), (b) and (c), the structures are as follows:

According to the above method, the prepared quinazoline compounds (a), (b) and (c) were tested for the biological activity of human lung cancer cell line A-549, and the test results showed that the quinazoline compounds (a), (b) ) and (c) had no obvious inhibitory effect on human lung cancer cell line A-549. Compounds (a), (b) and (c) had far less anticancer activity on human lung cancer cell line A-549 than compound (I). The specific results are shown in Table 2:

Table 2. Inhibitory effect of compounds (a), (b) and (c) on the growth of cancer cell line A-549

The above-mentioned anticancer activity in vitro test experiments showed that the other three compounds (a), (b) and (c) with similar structures had no obvious inhibitory effect on the growth of human lung cancer cell line A-549. Compound (I) has a significant inhibitory effect on the growth of human lung cancer cell line A-549, which is significantly better than that of compounds (a), (b) and (c).

(3) 4-chloroquinazoline was prepared by referring to the method in the literature (Rao, G.-W.et al.ChemMedChem, 2013, 8(6), 928-933), and then 4-chloroquinazoline was prepared according to Example 1. 6-nitroquinazoline was replaced with 4-chloroquinazoline, and other operations were the same as in Example 1, and the quinazoline compound (d) was synthesized, and the structure was as follows:

According to the above method, the prepared quinazoline compound (d) was tested for the biological activity of human lung cancer cell line A-549, and the test results showed that the quinazoline compound (d) had an anticancer effect on human lung cancer cell line A-549. The activity is far inferior to compound (I). The specific results are shown in Table 3:

Table 3. Inhibitory effect of compound (d) on the growth of cancer cell line A-549

(4) According to Example 11, pivaloyl chloride was replaced with benzoyl chloride, chloroacetyl chloride or isobutyryl chloride respectively, and other operations were the same as in Example 11, and quinazoline compounds (e), (h) and (j), the structure is as follows:

According to the above method, the prepared quinazoline compounds (e), (h) and (j) were tested for the biological activity of human lung cancer cell line A-549, and the test results showed that the quinazoline compounds (e), (h) ) and (j) have less anticancer activity than compound (I) on human lung cancer cell line A-549. The specific results are shown in Table 4:

Table 4. Inhibitory effect of compounds (e), (h) and (j) on the growth of cancer cell line A-549

Claims (2)

1. Pivalamidomethoxyphenyl benzo[d]azepine represented by formula (I)

Application of quinazoline compounds in the preparation of medicaments for preventing or treating human lung cancer;

2 . The application according to claim 1 , wherein the medicine is a medicine having the activity of inhibiting human lung cancer cell line A-549. 3 .