CN108245520B - Application of acetamidoquinazoline compounds in the preparation of medicaments for the treatment of lung cancer - Google Patents

Abstract

The invention discloses the application of an acetamidoquinazoline compound in the preparation of a drug for preventing or treating human lung cancer. The invention provides a novel quinazole with good anticancer (especially human lung cancer) activity The quinazoline compound is expected to be used in the preparation of medicines for preventing or treating human lung cancer; the preparation method of the acetamidoquinazoline compound provided by the present invention is simple and easy to operate, the raw materials are readily available, and the production cost is low, and is suitable for practical use.

Description

Application of acetamidoquinazoline compounds in the preparation of medicaments for the treatment of lung cancer

(1) Technical field

The invention relates to the application of an acetamidoquinazoline compound in the preparation of a medicament for preventing or treating tumor diseases.

(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

The object of the present invention is to provide a new type of quinazoline compound-acetamidoquinazoline compound in the preparation of medicine for preventing or treating human lung cancer, and the compound has a significant effect on human lung cancer cell line A-549 under a certain dose. In addition, the preparation method of this type of compound is simple, easy to operate, easy to obtain raw materials, and low in production cost, and is suitable for industrial application.

The technical scheme adopted in the present invention is:

The invention provides an application of an acetamidoquinazoline compound represented by formula (I) in the preparation of a medicine for preventing or treating human lung cancer,

Further, it is preferred that the drug is a drug that has the activity of inhibiting human lung cancer cell line A-549.

The preparation method of the acetamidoquinazoline compound represented by the formula (I) of the present invention is as follows: (1) mixing the compound represented by the formula (II) and the compound represented by the formula (III), in an organic solvent A, Under the action of the basic catalyst B, the reaction is carried out at 25～120°C (TLC tracking monitoring, the developing solvent is ethyl acetate/petroleum ether=1:3 (v/v), preferably 40～100°C for 0.5～12h), After the reaction is complete, the reaction solution is separated and purified to obtain a 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; the basic catalyst B is selected from one of the following: pyridine, diethylamine, triethylamine, quinoline, N,N-xylidine, 4-dimethylaminopyridine, 4 -pyrrolidinopyridine or sodium carbonate (preferably pyridine, diethylamine, triethylamine, N,N-dimethylaminopyridine or 4-dimethylaminopyridine); the compound represented by the formula (III) and the formula (II) The ratio of the amount of the compound shown to the basic catalyst B is 1.0:0.8～1.2:1.0～8.0, and the amount of the organic solvent A is 10～50mL/g in terms of the mass of the compound shown in formula (III) ;

(2) The compound represented by formula (IV) is in organic solvent D, under the action of reducing agent E, at 25～100 ℃, the reaction is complete (TLC tracking monitoring, 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 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 organic solvent D is one of the following: chloroform, toluene, methanol, ethanol, propylene alcohol, isopropanol, acetonitrile or N,N-dimethylformamide; when the reducing agent E is iron powder/concentrated hydrochloric acid or iron powder/acetic acid, the compound represented by formula (IV) and the reducing agent E The mass ratio of iron powder, concentrated hydrochloric acid or acetic acid is 1.0:1.0～3.0:0.2～1.0; When described reducing agent E is palladium carbon/ammonium formate or palladium carbon/hydrazine hydrate, formula (IV) The mass ratio of the compound shown and the palladium carbon, ammonium formate or hydrazine hydrate in the reducing agent E is 1.0:0.1～0.5:1.0～3.0; the consumption of the organic solvent D is the quality of the compound shown in formula (IV) Calculated as 10～50mL/g; the concentrated hydrochloric acid mass concentration of the present invention is 36%～38%, and the acetic acid adopts glacial acetic acid; the mass loading of palladium in the applicable palladium carbon in the present invention is 2～10%, preferably 5%, hydrated The mass concentration of hydrazine is 40-80%, preferably 80%;

(3) Mix the compound represented by formula (V) with acetyl chloride or acetic anhydride, under the action of basic catalyst F, in organic solvent G, the reaction is complete at -10 ~ 50 ℃ (TLC tracking monitoring, developing agent is ethyl acetate Ester/petroleum ether=1:1 (v/v), preferably -10～50℃ for 3～12h), the reaction solution is separated and purified 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 One: tetrahydrofuran, dichloromethane, chloroform, ethyl acetate, ether, acetonitrile, toluene or benzene; the ratio of the amount of the feed material of the compound shown in the described formula (V) and acetyl chloride or acetic anhydride, basic catalyst F The ratio is 1:1.0-8.0:1.0-3.0, and 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 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. Column chromatography silica gel (preferably 300-400 mesh coarse-pore (zcx.II) type column chromatography silica gel) is added with 1.0 to 2.0 times the weight of the concentrate, and after mixing, the solvent is evaporated and dried to obtain a mixture of the concentrate and silica gel , the mixture is packed into a column, and then the mixed solution of petroleum ether and ethyl acetate with a volume ratio of 1:0.1 to 10 is used as the eluent, and the effluent containing the target component is collected (preferably with ethyl acetate/petroleum ether=1: 3 (v/v) is the tracking detection of the developing agent, collecting target components, preferably collecting components with an Rf value of 0.5), concentrating under reduced pressure, and drying (preferably drying 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.

The specific recommended step (3) of the present invention is carried out according to the following method: under the condition of -10～10℃, the compound represented by formula (V) and the organic solvent G solution of basic catalyst F or the compound represented by formula (V) and The organic solvent G solution of acetyl chloride or acetic anhydride is added dropwise to the basic catalyst F, and the dropping is completed, and the reaction is carried out at -10 to 50 ° C for 3 to 12 hours, filtered, the filtrate is evaporated to remove the solvent, and the concentrate is obtained by column chromatography to obtain the formula (I). The volumetric dosage of the organic solvent for dissolving acetyl chloride or acetic anhydride has no effect on the present invention, the total dosage of the organic solvent G is 11-100 mL/g in terms of the mass of the compound shown in formula (V), and the total dosage of the organic solvent G is 11 to 100 mL/g. It refers to the total volume of the organic solvent for dissolving the catalyst and the compound represented by the formula (V) and the organic solvent for dissolving acetyl chloride or acetic 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 the concentrate into the liquid column chromatography silica gel (preferably 300 to 400 mesh coarse pore (zcx.II) type column chromatography silica gel), after mixing, evaporate the solvent and dry to obtain the concentrate and silica gel The mixture, the mixture is packed into a column, then the mixed solution of petroleum ether and ethyl acetate with a volume ratio of 1:0.1 to 10 is used as the eluent, and the effluent containing the target component is collected (preferably with ethyl acetate/petroleum ether= 1:1 (v/v) is the tracking detection of the developing agent, collect the target components, preferably collect the components with an Rf value of 0.5), concentrate under reduced pressure, dry (preferably dry at 50 ° C), and obtain the formula (I) shown in compound; the organic solvent H is one of the following: ethanol, chloroform, tetrahydrofuran or ethyl acetate; the amount of the organic solvent H is enough 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.

The acetamidoquinazoline (I) of the present invention has a significant inhibitory rate on human lung cancer cell line MCF-7, and can be used for preparing medicines for preventing or treating human lung cancer.

The beneficial effects of the present invention are mainly reflected in: (1) novel quinazoline compounds are provided, which have significant inhibitory activity against human lung cancer and are expected to be used in the preparation of medicines for preventing or treating human lung cancer; (2) the present invention provides The preparation method of the acetamidoquinazoline compound (I) is simple and easy to operate, the raw materials are readily available, and the production cost is low, which is suitable for practical use.

(4) Specific implementations

The present invention is further described below in conjunction with specific embodiment, but the protection scope of the present invention is not limited to this:

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 eluate is concentrated, and dried at 50 °C to obtain the eluate shown in formula (IV) The light yellow solid product was obtained, the yield was 85.1%, and the melting point was 164-166 °C. 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 eluate is concentrated, and dried at 50 °C to obtain the eluate shown in formula (IV) The light yellow solid product was obtained, the yield was 72.6%, and the melting point was 164-166 °C. 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 eluate is concentrated, and dried at 50 °C to obtain the eluate shown in formula (IV) The light yellow solid product was obtained, the yield was 77.2%, and the melting point was 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), the collected eluate is concentrated, and dried at 50 ° C to obtain the formula ( The light yellow solid product shown in IV) has a yield of 80.2% and a melting point of 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)), according to TLC detection, collect the eluate containing the compound represented by formula (IV) (Rf value is 0.5), the collected eluate Concentrate and dry at 50°C to obtain a pale yellow solid product represented by formula (IV), the yield is 89.6%, and the melting point is 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 represented by formula (IV) (Rf value is 0.5), concentrate the collected eluate, and dry at 50 °C to obtain the compound represented by formula (IV). The light yellow solid product was shown, the yield was 78.3%, and the melting point was 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 are the same

Example 7.

Example 11: Preparation of acetamidoquinazoline (I)

0.27 g (0.55 mmol) of 6-aminoquinazoline (V) prepared by the method in Example 7, 0.13 g (1.64 mmol) of pyridine, and 3 ml of tetrahydrofuran were successively added to the reaction flask, and 0.345 g of tetrahydrofuran was added dropwise under stirring at -10 °C. g (4.39mmol) acetyl 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 with 10 It was dissolved in milliliter 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 dissolving solution, and after mixing, the solvent was evaporated to obtain a mixture of dry concentrate and silica gel. mixture, the mixture is packed into a column, and then take 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=1:1 ( v/v)), collect the eluate containing the compound shown in formula (I) according to TLC detection (Rf value is 0.5), the collected eluate is concentrated, and dried at 50°C to obtain the acetamido group shown in formula (I). Quinazoline white solid, yield 67.6%, melting point 190～194℃. ¹ H NMR (500 MHz, CDCl ₃ ) δ: 2.27 (s, 3H), 3.26-3.33 (m, 1H), 3.52 (dt, J=15.3, 3.6 Hz, 1H), 3.75 (s, 3H), 3.79- 3.84(m,7H),3.95-4.07(m,2H),4.65(dd,J=7.9,14.1Hz,1H),5.24(t,J=8.7Hz,1H),6.65(s,1H),6.88 (d, J=8.7Hz, 2H), 7.08 (d, J=8.7Hz, 2H), 7.48 (d, J=8.9Hz, 1H), 7.65 (s, 1H), 7.81 (d, J=8.9Hz) , 1H), 8.57(s, 1H), 8.70(s, 1H). IR(KBr,cm ^-1 )ν: 3274, 2936, 1682, 1562, 1525, 1509, 1450, 1350, 1245, 1034,837.

Example 12: Preparation of acetamidoquinazoline (I)

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. 0.043 g (0.55 mmol) of acetyl 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)), and the reaction was carried out at 10°C. 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, 0.26 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 1:5 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, collect the eluate containing the compound shown in formula (I) (Rf value is 0.5), the collected eluate is concentrated, 50 After drying at °C, acetamidoquinazoline represented by formula (I) was obtained as a white solid with a yield of 82.3% and a melting point of 190-194 °C. ¹ H NMR and IR were the same as in Example 11.

Example 13: Preparation of acetamidoquinazoline (I)

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. 0.086 g (1.10 mmol) of acetyl 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 for 6 hours at 25°C. , filtered, the filtrate was evaporated to remove the solvent, the concentrate was dissolved in 20 ml of chloroform 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 Solvent, obtain the mixture of dry concentrate and silica gel, pack the mixture into column, then take the petroleum ether/ethyl acetate mixed solution whose volume ratio is 10:1 as eluent, elute, and TLC tracking detection (developing solvent is acetic acid) Ethyl ester/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), the collected eluate is concentrated and dried at 50°C The acetamidoquinazoline represented by the formula (I) was obtained as a white solid with a yield of 88.7% and a melting point of 190-194°C. ¹ HNMR and IR were the same as in Example 11.

Example 14: Preparation of acetamidoquinazoline (I)

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 0.224 g (2.19 mmol) of acetic 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), 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, after mixing, evaporated Remove the solvent to obtain a mixture of dry concentrate and silica gel, pack the mixture into a column, then use the petroleum ether/ethyl acetate mixed solution with a volume ratio of 5:1 as the eluent, elute, 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 represented by formula (I) (Rf value is 0.5), the collected eluate is concentrated, 50 ℃ After drying, the acetamidoquinazoline represented by the formula (I) was obtained as a white solid with a yield of 74.2% and a melting point of 190-194°C. ¹ H NMR and IR were the same as in Example 11.

Example 15: Preparation of acetamidoquinazoline (I)

0.27 g (0.55 mmol) of 6-aminoquinazoline (V), 0.213 g (1.65 mmol) of quinoline, and 15.0 ml of benzene prepared by the method of Example 10 were successively added to a 50-ml reaction flask, and stirred at -10°C. A solution of 0.173 g (2.20 mmol) of acetyl 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), reacted at -10°C for 12 hours, and filtered. , the filtrate was evaporated to remove the solvent, the concentrate was dissolved by adding 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, The mixture of dry concentrate and silica gel was obtained, and the mixture was packed into a column, and then took the petroleum ether/ethyl acetate mixed solution with a volume ratio of 1:1 as the eluent, elution, and TLC tracking 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), the collected eluate is concentrated, and dried at 50°C to obtain the formula The acetamidoquinazoline shown in (I) was a white solid, the yield was 62.5%, and the melting point was 190-194°C. ¹ H NMR and IR were the same as in Example 11.

Example 16: Preparation of acetamidoquinazoline (I)

0.27 g (0.55 mmol) of 6-aminoquinazoline (V) prepared by the method of Example 9, 0.164 g (1.10 mmol) of 4-pyrrolidinopyridine, and 15.0 ml of dichloromethane were successively added to a 50 ml reaction flask , 0.086 g (1.10 mmol) of acetyl chloride and 5.0 ml of dichloromethane solution were added dropwise under stirring at 10 °C, and the dropping was completed, TLC tracking detection (developing solvent was ethyl acetate/petroleum ether=1:1), under 10 °C React for 8 hours, filter, evaporate the solvent from the filtrate, add 20 ml of ethanol to the concentrate to dissolve it to obtain a dissolving solution, add 0.50 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the dissolving solution, and mix well. , 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 10:1 as the eluent, elution, and TLC tracking detection (develop The agent 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 eluate is concentrated, After drying at 50°C, the acetamidoquinazoline represented by formula (I) was obtained as a white solid with a yield of 79.8% and a melting point of 190-194°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.

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

Cell line: human lung cancer cell line A-549, purchased from the Cell Bank of Shanghai Academy of Biological Sciences, Chinese Academy of Sciences.

The experimental steps are as follows:

(a) Preparation of samples: For soluble samples, dissolve with 40 μL DMSO per 1 mg, 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 10th 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, add 100 μg/mL, 10 μg/mL and 1 μg/mL samples diluted with culture medium, 100 μL per well, add 3 wells for each concentration, and incubate at 37°C in a 5% CO ₂ incubator for 72 h. 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 lung cancer cell line A-549

(2) according to embodiment 11, acetyl chloride is replaced with 4-iodobenzoyl chloride, 3-methoxybenzoyl chloride and cinnamoyl chloride respectively, and other operations are the same as in embodiment 11, respectively synthesizing quinazoline compounds (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) have no obvious inhibitory effect on human lung cancer cell line A-549, and the anticancer activities of compounds (a), (b) and (c) on human lung cancer cell line A-549 are far less than that of acetamidoquinazoline. (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, the acetyl chloride was replaced with benzoyl chloride, butyryl chloride, propionyl chloride or chloroacetyl chloride respectively, and other operations were the same as in Example 11, and quinazoline compounds (e) and (f) were synthesized respectively. , (g) and (h), the structures are as follows:

According to the above method, the prepared quinazoline compounds (e), (f), (g) and (h) were tested for biological activity of human lung cancer cell line A-549, and the test results showed that the quinazoline compounds (e) ), (f), (g) and (h) had 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), (f), (g) and (h) on the growth of cancer cell line A-549

Claims (2)

1. an application of an acetamidoquinazoline compound shown in formula (I) in the preparation of a medicine 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 the human lung cancer cell line A-549. 3 .