CN108014113A - Application of butyrylamidodimethoxybenzo [ d ] aza-based quinazoline compound in preparation of drugs for treating cervical cancer - Google Patents

Abstract

The invention discloses a butyrylaminodimethoxybenzo[d]azepine The invention discloses an application of quinazoline compounds in the preparation of drugs for preventing or treating human cervical cancer, and the quinazoline compounds have a significant effect on inhibiting the Siha activity of human cervical cancer cell lines.

Description

Butyramide dimethoxy benzo [d] aza * base quinazoline compounds in the preparation Application of drugs in the treatment of cervical cancer

(1) Technical field

The present invention relates to a kind of butyramide dimethoxy benzo [d] azepine Application of quinazoline compounds in the preparation of medicines 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., quinazoline Some compounds have been listed 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, all belong to 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 kind of novel quinazoline compound - butyrylamino dimethoxy benzo [d] azepine The application of quinazoline compounds in the preparation of drugs for preventing or treating human cervical cancer, the compounds have a significant inhibitory rate to the human cervical cancer cell line Siha at a certain dose; and the preparation method of the compounds is simple and easy operation, the raw materials are easy to obtain, and the production cost is relatively low, which is suitable for industrial application.

In order to realize the above-mentioned purpose of the invention, the present invention adopts following technical scheme:

The invention provides a butyramide dimethoxy benzo [d] azepine shown in formula (I) The application of quinazoline compounds in the preparation of drugs for the prevention or treatment of tumors, especially the application in the preparation of drugs for the prevention or treatment of human cervical cancer:

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

The present invention provides a butyramide dimethoxy benzo [d] azepine represented by formula (I) The preparation method of quinazoline compound, described method is: (1) compound shown in formula (II) is mixed with the compound shown in formula (III), in organic solvent A, in the effect of basic catalyst B 25～120℃ for reaction (TLC tracking monitoring, developer 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 shown in 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-dimethylaniline, 4-dimethylaminopyridine, 4-pyrrolidinylpyridine or carbonic acid Sodium (preferably pyridine, diethylamine, triethylamine, N,N-dimethylaniline or 4-dimethylaminopyridine);

(2) Dissolve the compound represented by formula (IV) obtained in step (1) in organic solvent D, and react completely at 25 to 100°C under the action of reducing agent E (TLC tracking monitoring, developing agent is ethyl acetate/ Petroleum ether = 1:1 (v/v), preferably at 40-80°C for 0.5-12h), filter the reaction solution, and dry the concentrate after the filtrate is concentrated under reduced pressure (preferably vacuum-dried at 25°C) to obtain formula (Ⅴ) The compound shown; 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 One: 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, iron powder/acetic acid refers to The mixing of iron powder and acetic acid in any proportion, the palladium carbon/ammonium formate refers to the mixing of palladium carbon and ammonium formate in any proportion, and the palladium carbon/hydrazine hydrate is a mixture of palladium carbon and hydrazine hydrate in any proportion;

(3) Mix the compound shown in the formula (Ⅴ) obtained in step (2) with butyryl chloride or butyric anhydride, under the action of basic catalyst F, in organic solvent G, the reaction is complete at -10～50°C (TLC tracking monitoring, The developing agent is ethyl acetate/petroleum ether=1:1 (v/v), preferably -10～50°C for 3～12h), and the reaction solution is post-treated to obtain the compound shown in formula (I); The organic solvent G is one of the following: tetrahydrofuran, dichloromethane, chloroform, ethyl acetate, ether, acetonitrile, toluene or benzene; the basic catalyst F is one of the following: pyridine, diethylamine, triethylamine, Quinoline, N,N-xylidine, 4-dimethylaminopyridine, 4-pyrrolidinylpyridine or sodium carbonate;

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 basic catalyst B 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 based on the mass of the compound represented by formula (III).

Further, the method for separating and purifying the reaction solution in step (1) of the present invention is: after the reaction is complete, the reaction solution is evaporated to remove the solvent, and the concentrate is dissolved with an organic solvent C 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 on column chromatography silica gel (preferably 300 to 400 mesh coarse-pore (zcx.II) type column chromatography silica gel), mix well, evaporate the solvent, and dry to obtain a mixture of the concentrate and silica gel , put the mixture into a column, and then use a mixed solution of petroleum ether and ethyl acetate with a volume ratio of 1:0.1 to 10 as the eluent to collect the effluent containing the target component (preferably with ethyl acetate/petroleum ether=1: 3 (v/v) is developer tracking detection, collect target components, preferably collect components with Rf value of 0.5), concentrate under reduced pressure, and dry (preferably 50°C drying), to obtain the compound shown in 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), the reducing agent E is iron powder/concentrated hydrochloric acid or iron powder/acetic acid, the feed quality of the compound shown in formula (IV) and the iron powder, concentrated hydrochloric acid or acetic acid in reducing agent E The ratio is 1.0:1.0~3.0:0.2~1.0. The mass concentration of concentrated hydrochloric acid in the present invention is 36%-38%, and the acetic acid is glacial acetic acid.

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

Further, in step (2), the amount of the organic solvent D used is 10-50 mL/g based on 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 butyryl chloride or butyric anhydride, and the 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 based on the mass of the compound represented by formula (V).

Further, step (3) is carried out according to the following method: under the condition of -10～10°C, add the compound shown in formula (Ⅴ) and the organic solvent G solution of basic catalyst F or add the compound shown in formula (Ⅴ) and the base The organic solvent G solution of butyryl chloride or butyric anhydride is added dropwise to the catalyst F, and the dropwise reaction is carried out at -10 to 50° C. for 3 to 12 hours, and the resulting reaction solution is post-treated to obtain the compound shown in formula (I); Or the organic solvent volume consumption of butyric anhydride has no influence on the present invention, and the total consumption of described organic solvent G is 11～100mL/g by the mass of the compound shown in formula (V). The total consumption of organic solvent G refers to the total volume of the organic solvent G that dissolves the basic catalyst F and the compound shown in formula (Ⅴ) and the organic solvent G that dissolves butyryl chloride or butyric anhydride.

Further, the post-processing method of the reaction solution described in step (3) of the present invention is: filter the reaction solution, evaporate the filtrate to remove the solvent, take the concentrate and dissolve it with an organic solvent H to obtain a solution, and then add concentrated 1.0 to 2.0 times the weight of 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 a mixture of concentrate and silica gel. The mixture is packed into a column, and then a mixed solution of petroleum ether and ethyl acetate with a volume ratio of 1:0.1 to 10 is used as an eluent to collect the effluent containing the target component (preferably with ethyl acetate/petroleum ether=1:1( v/v) is a developing agent tracking detection, collecting target components, preferably collecting components with an Rf value of 0.5), concentrating under reduced pressure, drying (preferably drying at 50° C.), and obtaining the compound shown in formula (I); 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, named for the convenience of different organic solvents used in different steps, and the letters themselves have no meaning; the catalyst B, reducing agent E and catalyst F are Catalysts are named in order to distinguish the different catalysts used in different steps, and the letters themselves have no meaning.

The beneficial effect of the present invention is mainly embodied in that it provides the application of a novel quinazoline compound in the preparation of medicines for preventing or treating human cervical cancer, and the compound has significant inhibitory activity on human cervical cancer cell line Siha.

(4) Specific implementation methods

The present invention is further described in conjunction with specific examples, and the following examples illustrate the present invention, rather than limit the present invention in any way.

Compound (II) was prepared according to the method in the literature (Weinstock, J. et al. J. Med. Chem., 1986, 29(11), 2315-2325). 4-Chloro-6-nitroquinazoline (Ⅲ) was prepared 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 that the embodiment of the present invention uses: D5H5A, manufacturer: Shaanxi Ruike New Material Co., Ltd.

Example 1: Nitrobenzo[d]azepines The preparation of quinazoline (IV)

1.20 grams (5.73mmol) of 4-chloro-6-nitroquinazoline (Ⅲ) and 2.39 grams (6.87mmol) of compound (II), 3.62 grams (45.76mmol) of pyridine, and 12 milliliters of chloroform were added to 50 milliliters of the reaction In the bottle, heated to 40°C, followed by TLC detection (developing solvent: ethyl acetate/petroleum ether = 1:3 (v/v)), stirred for 10 hours, closed the reaction, evaporated the reaction solution to remove the solvent, and obtained the concentrate 10 milliliters of ethyl acetate was added to dissolve it to obtain a solution, and 3.0 grams of column chromatography silica gel (300-400 mesh column chromatography silica gel) was added to the solution, and after mixing, the solvent was evaporated to obtain a dry concentrate The mixture with silica gel, the mixture was packed into a column, and then the mixed solution of petroleum ether/ethyl acetate with a volume ratio of 1:10 was used as the eluent for elution, and 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 eluate is concentrated, dried at 50°C to obtain the compound shown in formula (IV) The light yellow solid product, yield 85.1%, melting point 164 ~ 166 ℃. ¹ H NMR (500MHz, CDCl ₃ ) δ: 3.32-3.38 (m, 1H), 3.63 (dt, J=3.4, 15.5Hz, 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 =9.1Hz, 1H), 8.48(dd, J=2.4, 9.2Hz, 1H), 8.71(s, 1H), 8.96(d, J=2.4Hz, 1H). IR (KBr, cm ^-1 ) ν: 2917, 2848, 1616, 1580, 1510, 1463, 1355, 1327, 1249, 1038, 847.

Example 2: Nitrobenzo[d]azepines The preparation of quinazoline (IV)

1.20 grams (5.73 mmol) of 4-chloro-6-nitroquinazoline (Ⅲ) and 1.59 grams (4.57 mmol) of compound (II), 1.67 grams (22.83 mmol) of diethylamine, and 60 milliliters of toluene were added to 100 milliliters successively. In a three-neck flask, heated to 100°C, followed by TLC detection (developing solvent is ethyl acetate/petroleum ether=1:3 (v/v)), stirred for 2 hours, closed the reaction, and evaporated the solvent from the reaction solution to obtain Add 20 ml of ethanol to the concentrate to dissolve it to obtain a solution, add 2.5 grams of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the solution, mix well, and evaporate the solvent to obtain a dry concentrate With the mixture of silica gel, the mixture is packed into a column, and then the mixed solution of petroleum ether/ethyl acetate with a volume ratio of 1:5 is used as the eluent for 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 compound shown in formula (IV) The light yellow solid product, yield 72.6%, melting point 164 ~ 166 ℃. ¹ H NMR and IR are the same as in Example 1.

Example 3: Nitrobenzo[d]azepines The preparation of quinazoline (IV)

1.20 grams (5.73 mmol) of 4-chloro-6-nitroquinazoline (Ⅲ) and 1.99 grams (5.72 mmol) of compound (II), 0.58 grams (5.73 mmol) of triethylamine, and 60 milliliters of ethanol were added to 100 milliliters of In a three-necked flask, heated to 60°C, followed by TLC detection (developing solvent is ethyl acetate/petroleum ether=1:3 (v/v)), stirred for 8 hours, closed the reaction, and evaporated the solvent from the reaction solution to obtain Add 20 ml of chloroform to the concentrate to dissolve it to obtain a solution, add 2.5 grams of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the solution, mix well, and evaporate the solvent to obtain a dry concentrate With the mixture of silica gel, the mixture is packed into a column, and then the mixed solution of petroleum ether/ethyl acetate with a volume ratio of 10:1 is used as the eluent for 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 compound shown in formula (IV) The light yellow solid product, yield 77.2%, melting point 164 ~ 166 ℃. ¹ H NMR and IR are the same as in Example 1.

Example 4: Nitrobenzo[d]azepines The preparation of quinazoline (IV)

1.20 grams (5.73mmol) of 4-chloro-6-nitroquinazoline (Ⅲ) and 2.20 grams (6.32mmol) of compound (II), 1.40 grams (11.46mmol) of 4-dimethylaminopyridine, and 60 milliliters of isoquinazoline were successively Add propanol to a 100 ml three-neck flask, stir at room temperature 25°C, track and detect with TLC (developing solvent is ethyl acetate/petroleum ether = 1:3 (v/v)), react for 12 hours, close the reaction, and evaporate the reaction liquid Solvent, add 20 milliliters of tetrahydrofuran to the obtained concentrate to dissolve it, obtain a solution, add 4.0 grams of column chromatography silica gel (300 ~ 400 mesh column chromatography silica gel) to the solution, after mixing, evaporate the solvent to obtain The mixture of the dried concentrate and silica gel was packed into a column, and then the mixed solution of petroleum ether/ethyl acetate with a volume ratio of 5:1 was used as the eluent for elution, and TLC tracking detection (developing solvent was ethyl acetate/ethyl acetate). Petroleum ether=1: 3 (v/v)), detect and collect the eluate (Rf value 0.5) containing the compound shown in formula (IV) according to TLC, the eluate collected is concentrated, and 50 ℃ of dryings obtain formula ( The light yellow solid product shown in IV) has a yield of 80.2% and a melting point of 164-166°C. ¹ HNMR and IR are the same as in Example 1.

Example 5: Nitrobenzo[d]azepines The preparation of quinazoline (IV)

1.20 grams (5.73mmol) of 4-chloro-6-nitroquinazoline (Ⅲ) and 1.79 grams (5.15mmol) of compound (II), 1.04 grams (8.58mmol) of N,N-xylidine, 12 ml Add N,N-dimethylformamide into a 50ml reaction flask, heat to 120°C, track and detect with TLC (developing solvent is ethyl acetate/petroleum ether=1:3 (v/v)), and stir for 0.5 hours , close the reaction, the reaction solution is evaporated to remove the solvent, and 20 milliliters of tetrahydrofuran is added to the obtained concentrate to dissolve it to obtain a solution, and 5.0 grams of column chromatography silica gel (300-400 mesh column chromatography silica gel) are added to the solution, and mixed After homogenization, the solvent was evaporated to obtain a mixture of the dry concentrate and silica gel, and the mixture was packed into a column, and then the mixed solution of petroleum ether/ethyl acetate with a volume ratio of 1:1 was used as the eluent for elution, and TLC tracking detection (developing agent is ethyl acetate/petroleum ether=1:3 (v/v)), detects and collects the eluate containing the compound shown in formula (IV) according to TLC (Rf value is 0.5), the eluate collected Concentrate and dry at 50°C to obtain a pale yellow solid product represented by formula (IV), with a yield of 89.6% and a melting point of 164-166°C. ¹ H NMR and IR are the same as in Example 1.

Example 6: Nitrobenzo[d]azepines The preparation of quinazoline (IV)

1.20 grams (5.73mmol) of 4-chloro-6-nitroquinazoline (Ⅲ) and 2.39 grams (6.87mmol) of compound (II), 3.62 grams (45.76mmol) of pyridine, and 20 milliliters of propanol were added to 50 milliliters of In the reaction flask, heated to 40°C, followed by TLC detection (developing solvent: ethyl acetate/petroleum ether = 1:3 (v/v)), stirred for 10 hours, closed the reaction, evaporated the reaction solution to remove the solvent, and concentrated 20 milliliters of ethyl acetate was added to the mixture to dissolve it to obtain a solution, and 3.5 grams of column chromatography silica gel (300-400 mesh column chromatography silica gel) was added to the solution, and after mixing, the solvent was evaporated to obtain dry concentrated The mixture of compound and silica gel, the mixture was packed into a column, and then the mixed solution of petroleum ether/ethyl acetate with a volume ratio of 1:1 was used as eluent, eluted, and 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 eluate is concentrated, and dried at 50°C to obtain the compound of formula (IV). The pale yellow solid product shown, the yield is 78.3%, and the melting point is 164-166°C. ¹ H NMR and IR are the same as in Example 1.

Example 7: Aminobenzo[d]azepines Preparation of Quinazoline (Ⅴ)

0.40 gram (0.77mmol) nitrobenzo [d] azepines prepared by the method of Example 1 were successively Quinazoline (IV), 0.40 gram (6.34mmol) ammonium formate, 0.04 gram 5%Pd/C, 4.0 milliliters of chloroform joins in the reaction flask, stirs at room temperature 25 ℃, TLC tracking detects (developing agent is ethyl acetate/ Petroleum ether = 1:1 (v/v)), reacted for 12 hours, filtered, concentrated the filtrate, and dried under vacuum at 25°C to obtain a light yellow solid product, aminobenzo[d]azepine Quinazoline (Ⅴ) with a yield of 98.2% and a melting point of 122-126°C. 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: Aminobenzo[d]azepines Preparation of Quinazoline (Ⅴ)

0.40 gram (0.77mmol) nitrobenzo[d]azepines prepared by the method of Example 2 were successively Quinazoline (IV), 1.20 gram (19.18mmol) 80wt% hydrazine hydrate, 0.20 gram 5%Pd/C, 20.0 milliliters of toluene join in the reaction bottle of 50 milliliters, be heated to 100 ℃, TLC follow-up detection (developing solvent Ethyl acetate/petroleum ether=1:1 (v/v)), stirred for 0.5 hours, cooled and filtered, the filtrate was concentrated, and dried in vacuum at 25°C to obtain a light yellow solid product, aminobenzo[d]azepine Quinazoline (V), yield 100.0%, melting point 122-126°C. ¹ H NMR and IR are the same as in Example 7.

Example 9: Aminobenzo[d]azepines Preparation of Quinazoline (Ⅴ)

0.40 gram (0.77mmol) nitrobenzo [d] azepines prepared by the method of Example 3 were successively Quinazoline (IV), 0.08 gram of concentrated hydrochloric acid (mass concentration 36～38%), 0.40 gram of iron powder, 20.0 milliliters of methyl alcohol join in the reaction bottle of 50 milliliters, be heated to 40 ℃, TLC follow-up detection (developing agent is Ethyl acetate/petroleum ether=1:1 (v/v)), stirred for 8 hours, cooled and filtered, the filtrate was concentrated, and vacuum-dried at 25°C to obtain a pale yellow solid product aminobenzo[d]azepine Quinazoline (Ⅴ) with a yield of 94.1% and a melting point of 122-126°C. ¹ HNMR and IR are the same as in Example 7.

Example 10: Aminobenzo[d]azepines Preparation of Quinazoline (Ⅴ)

0.40 gram (0.77mmol) nitrobenzo [d] azepines prepared by the method of embodiment 4 were successively Quinazoline (IV), 0.40 gram of acetic acid, 1.20 gram of iron powder, 20.0 milliliters of isopropanols join in the reaction bottle of 50 milliliters, be heated to 80 ℃, TLC tracking detection (developing agent is 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 a light yellow solid product aminobenzo[d]azepine Quinazoline (Ⅴ) with a yield of 97.5% and a melting point of 122-126°C. ¹ H NMR and IR are the same as in Example 7.

Example 11: Butyrylaminodimethoxybenzo[d]azepine The preparation of quinazoline (I)

0.27 grams (0.55 mmol) of aminobenz [d] azepines prepared by the method of Example 7 were successively Quinazoline (Ⅴ), 0.13 gram (1.64mmol) pyridine, 3 milliliters of tetrahydrofuran joins in the reaction flask, drips 0.469 gram (4.40mmol) butyryl chloride under the stirring condition of-10 ℃, dropwise finishes, and TLC tracking detects (developing The agent is 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 by adding 10 ml of ethyl acetate to obtain a solution, which was added to the solution Add 0.60 g of column chromatography silica gel (300-400 mesh column chromatography silica gel), mix well, and evaporate the solvent to obtain a mixture of dry concentrate and silica gel. Petroleum ether/ethyl acetate mixed solution is the eluent, elution, TLC follow-up detection (developing solvent is ethyl acetate/petroleum ether=1:1 (v/v)), according to TLC detection, collect the compound containing the compound of formula (I). The eluent of the compound shown (Rf value is 0.5), the collected solution is concentrated, and 50 ℃ of drying obtains the butyrylamino dimethoxy benzo [d] azepine shown in formula (I) Quinazoline is an off-white solid with a yield of 47.2% and a melting point of 216-217°C. ¹ H NMR (500MHz, CDCl ₃ )δ: 1.02(t, J=7.4Hz, 3H); 1.76-1.83(m, 2H); 2.41-2.51(m, 2H); 3.24-3.30(m, 1H), 3.54(dt,J=3.6,15.1Hz,1H),3.74(s,3H),3.81-3.82(m,7H),3.98-4.09(m,2H),4.66(dd,J=8.3,14.2Hz, 1H), 5.27(t, J=8.8Hz, 1H).6.67(s, 1H), 6.88(d, J=8.8Hz, 2H), 7.07(d, J=8.7Hz, 2H), 7.61(dd, J=2.0, 9.0Hz, 1H), 7.80(d, J=8.9Hz, 1H), 8.40(s, 1H), 8.53(s, 1H), 8.85(d, J=1.8Hz, 1H). HRMS-ESI m/z: 561.2265 [M+H] ⁺ . IR (KBr, cm ^-1 ) ν: 2960, 2933, 2870, 2835, 1692, 1562, 1523, 1511, 1488, 1463, 1349, 1250, 1035, 836.

Example 12: Butyrylaminodimethoxybenzo[d]azepine The preparation of quinazoline (I)

0.27 grams (0.55 mmol) of aminobenz [d] azepines prepared by the method of Example 8 were successively Quinazoline (Ⅴ), 0.04 gram (0.55 mmol) diethylamine, 10.0 milliliters of chloroform are added in the reaction bottle of 50 milliliters, 0.059 gram (0.55 mmol) butyryl chloride and 5.0 milliliters of chloroform are mixed dropwise under 10 ℃ of stirring conditions Solution, dropwise, TLC tracking detection (developing solvent: ethyl acetate/petroleum ether = 1:1 (v/v)), react at 10°C for 8 hours, filter, distill the filtrate to remove solvent, add 20 ml of ethanol to the concentrate Dissolve it to obtain a solution, add 0.26 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the solution, mix well, and evaporate the solvent to obtain a mixture of dry concentrate and silica gel. Pack the column, then use petroleum ether/ethyl acetate mixed solution with a volume ratio of 1:5 as eluent, elute, and TLC tracking detection (developing solvent is ethyl acetate/petroleum ether=1:1 (v/v) ), detect and collect the eluate containing the compound shown in formula (I) (Rf value is 0.5) according to TLC, collect liquid and concentrate, and dry at 50 ℃ to obtain the butyramide dimethoxy benzo shown in formula (I) [d] Aza Quinazoline is an off-white solid with a yield of 32.9% and a melting point of 216-217°C. ¹ H NMR and IR are the same as in Example 11.

Example 13: Butyrylaminodimethoxybenzo[d]azepine The preparation of quinazoline (I)

0.27 grams (0.55 mmol) of aminobenz [d] azepines prepared by the method of Example 9 were successively Quinazoline (Ⅴ), 0.111 gram (1.10mmol) triethylamine, 10.0 milliliters of ethyl acetate joins in the reaction bottle of 50 milliliters, drips 0.117 gram (1.10 mmol) butyryl chloride and 5.0 milliliters under stirring condition at 0 ℃ Ethyl acetate solution, dropwise, TLC follow-up detection (developer: ethyl acetate/petroleum ether = 1:1), react at 25°C for 6 hours, filter, distill off the solvent from the filtrate, add 20 ml of chloroform to the concentrate Dissolve to obtain a solution, add 0.30 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the solution, mix well, evaporate the solvent to obtain a mixture of dry concentrate and silica gel, and 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 to 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 shown in formula (I) (Rf value is 0.5), collect liquid concentration, 50 ℃ of drying obtain the butyrylamino dimethoxy benzo [d shown in formula (I) ] Aza Quinazoline is an off-white solid with a yield of 46.6% and a melting point of 216-217°C. ¹ H NMR and IR are the same as in Example 11.

Example 14: Butyrylaminodimethoxybenzo[d]azepine The preparation of quinazoline (I)

0.27 grams (0.55 mmol) of aminobenz [d] azepines prepared by the method of Example 10 were successively Quinazoline (Ⅴ), 0.067 gram (0.55mmol) 4-dimethylaminopyridine, 20.0 milliliters of toluene joins in the reaction flask of 50 milliliters, drips 0.348 gram (2.20mmol) butyric anhydride and 7.0 milliliters under 5 ℃ of stirring conditions A solution of one milliliter of toluene, after dropping, heated to 50°C, followed by TLC detection (developing solvent: ethyl acetate/petroleum ether = 1:1), reacted for 3 hours, filtered, the filtrate was distilled to remove the solvent, and the concentrate was added with 20 ml of tetrahydrofuran to dissolve It was dissolved to obtain a solution, and 0.40 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) was added to the solution, and after mixing, the solvent was evaporated to obtain a mixture of dry concentrate and silica gel, and the mixture was packed Column, then use petroleum ether/ethyl acetate mixed solution with a volume ratio of 5:1 as eluent, elute, TLC tracking detection (developing solvent is ethyl acetate/petroleum ether=1:1 (v/v)) According to TLC detection, collect the eluent containing the compound shown in formula (I) (Rf value is 0.5), collect the liquid and concentrate, dry at 50 DEG C to obtain the butyrylaminodimethoxybenzo[ d] aza Quinazoline is an off-white solid with a yield of 50.7% and a melting point of 216-217°C. ¹ H NMR and IR are the same as in Example 11.

Example 15: Butyrylaminodimethoxybenzo[d]azepine The preparation of quinazoline (I)

0.27 grams (0.55 mmol) of aminobenz [d] azepines prepared by the method of Example 7 were successively Quinazoline (Ⅴ), 0.213 gram (1.65mmol) quinoline, 15.0 milliliters of benzene join in the reaction bottle of 50 milliliters, under the stirring condition of-10 ℃, dropwise add 0.234 gram (2.20mmol) butyryl chloride and 5.0 milliliters of benzene Solution, dropwise, TLC follow-up detection (developing solvent: ethyl acetate/petroleum ether = 1:1), react at -10°C for 12 hours, filter, evaporate the filtrate to remove the solvent, add 20 ml of tetrahydrofuran to the concentrate to dissolve it, To obtain the solution, add 0.40 g of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the solution, mix well, evaporate the solvent to obtain a mixture of dry concentrate and silica gel, and pack the mixture into a column, then Use petroleum ether/ethyl acetate mixed solution with volume ratio of 1:1 as eluent, elute, TLC tracking detection (developing solvent is ethyl acetate/petroleum ether=1:1 (v/v)), according to TLC Detect and collect the eluent containing the compound represented by formula (I) (Rf value is 0.5), concentrate the collected liquid, and dry at 50°C to obtain butyrylaminodimethoxybenzo[d]nitrogen represented by formula (I) miscellaneous Quinazoline is an off-white solid with a yield of 51.4% and a melting point of 216-217°C. ¹ H NMR and IR are the same as in Example 11.

Example 16: Butyrylaminodimethoxybenzo[d]azepine The preparation of quinazoline (I)

0.27 grams (0.55 mmol) of aminobenz [d] azepines prepared by the method of Example 7 were successively Quinazoline (Ⅴ), 0.164 gram (1.10mmol) 4-pyrrolidinylpyridine, 15.0 milliliters of dichloromethane joins in the reaction bottle of 50 milliliters, drips 0.117 gram (1.10mmol) butyryl chloride under 10 ℃ of stirring conditions and 5.0 ml of dichloromethane solution, dropwise, TLC follow-up detection (developing solvent: ethyl acetate/petroleum ether = 1:1), react at 10°C for 8 hours, filter, distill the filtrate to remove solvent, add 20 ml of concentrate Dissolve it with ethanol to obtain a solution, add 0.50 gram of column chromatography silica gel (300-400 mesh column chromatography silica gel) to the solution, after mixing, evaporate the solvent to obtain a dry concentrate and a mixture of silica gel, The mixture was loaded 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 for elution, and TLC tracking detection (developing solvent was ethyl acetate/petroleum ether=1:1 (v/v )), detect and collect the eluate (Rf value is 0.5) containing the compound shown in formula (I) according to TLC, collect the liquid and concentrate, dry at 50 DEG C to obtain the butyramide dimethoxybenzene shown in formula (I) And[d]aza Quinazoline is an off-white solid with a yield of 43.7% and a melting point of 216-217°C. ¹ H NMR and IR are the same as in Example 11.

Example 17: In vitro test 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 Institute 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 culture medium: Each 1000 mL DMEM medium (Gibco) contains 800,000 units of penicillin, 1.0 g of streptomycin, and 10% inactivated fetal bovine serum.

②Cultivation of cells: Inoculate the tumor cells in the culture medium, culture them in a 37°C, 5% CO ₂ incubator, and passage them 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. 24 hours after inoculation, add 100 μg/mL, 10 μg/mL and 1 μg/mL samples diluted with culture medium respectively, 100 μL per well, add 3 wells for each concentration, culture in a 37°C, 5% _CO2 incubator, and after 72 hours 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 an oscillator to completely dissolve formazan, and use a microplate reader to perform colorimetry at a wavelength of 570 nm . The cells cultured in the medium containing no sample and the same concentration of DMSO under the same conditions were used as a control, and the IC ₅₀ of the sample on tumor cell growth was calculated.

The test results are shown in Table 1:

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

(2) According to Example 11, butyryl chloride is replaced with 3-methoxybenzoyl chloride or cinnamoyl chloride respectively, and other operations are the same as in Example 11, and quinazoline compounds (b) and (c) have been synthesized respectively. As follows:

The quinazoline compounds (b) and (c) prepared according to the above method have been tested for human cervical cancer cell line Siha biological activity, and the test results show that quinazoline compounds (b) and (c) are effective on human cervical cancer The inhibitory effects on cell line Siha were 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. Compounds (b) and (c) inhibit the growth of cancer cell line Siha

The in vitro test of the above anticancer activity showed that the other two compounds (b) and (c) with similar structures had 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 obviously better than compounds (b) and (c).

(3) According to embodiment 11, butyryl chloride is replaced with cyclohexyl methyl chloroformate, and other operations are the same as embodiment 11, and quinazoline compound (k) is synthesized, and the structure is as follows:

The quinazoline compounds (k) prepared according to the method described above have carried out the human cervical cancer cell line Siha biological activity test, test result shows that compound (k) is far inferior to compound (I) to the anticancer activity of human cervical cancer cell line Siha ). The specific results are shown in Table 3:

Table 3. The inhibitory effect of compound (k) on the growth of cancer cell line Siha

Claims (2)

1. Butyramide dimethoxy benzo [d] azepine shown in a kind of formula (I) Application of quinazoline compounds in the preparation of medicines for preventing or treating human cervical cancer: 2. The application according to claim 1, characterized in that: the drug is a drug that has the activity of inhibiting human cervical cancer cell line Siha.