CN108047206B - Pivaloyl amino benzo [d] azepine * base quinazoline compounds and preparation and application - Google Patents
Pivaloyl amino benzo [d] azepine * base quinazoline compounds and preparation and application Download PDFInfo
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- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
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Abstract
The invention discloses a kind of pivaloyl amino benzo [d] azepinesBase quinazoline compounds and preparation and application.Pivaloyl amino benzo [d] azepine provided by the inventionBase quinazoline compounds all have significant inhibitory activity to MCF-7 cell strainHJ2mm, human lung cancer cell lines A-549, people in loop strain HL-60 or human cervical carcinoma cell lines Siha, are expected to be applied in preparation prevention or treatment human breast carcinoma, human lung cancer, human leukemia or the drug of human cervical carcinoma.The present invention provides pivaloyl amino benzo [d] azepines
Description
(1) technical field
The present invention relates to a kind of quinazoline compounds and its application, in particular to a kind of pivaloyl amino benzo [d] nitrogen
It is miscellaneousBase quinazoline compounds and preparation method thereof and the compound are in preparation prevention or the drug for the treatment of tumor disease
In application.
(2) background technique
Quinazoline compounds have many preferable bioactivity, have a wide range of applications in field of medicaments, and especially one
The quinazoline derivative of a little special constructions has apparent antiviral activity, antibacterial activity, anti-tumor activity etc., quinazoline ditosylate salt
Compound has had listed some kinds as anti-tumor drug.Such as the Gefitinib for being used to treat lung cancer of listing
(Gefitinib) and Tarceva (Erlotinib), and the Lapatinib (Lapatinib) for treating breast cancer, they
Belong to quinazoline compounds.Novel quinazoline compounds and its bioactivity also common document report (refering to 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).Certainly majority quinazoline
Class compound does not simultaneously have anti-tumor activity.
(3) summary of the invention
The purpose of the present invention is to provide a kind of novel quinazoline quinoline class compound-pivaloyl amino with anticancer activity
Benzo [d] azepineBase quinazoline compounds and its preparation method and application, such compound is under doses to people's lung
Cancer cell line A-549, MCF-7 cell strainHJ2mm, people in loop strain HL-60 or human cervical carcinoma cell lines
Siha Zhuo has significant inhibiting rate；And such compounds process for production thereof is easy, easily operated, raw material is easy to get, and production cost
It is lower, it is suitable for industrial applications.
For achieving the above object, the present invention adopts the following technical scheme:
In a first aspect, the present invention provides pivaloyl amino benzo [d] azepines shown in a kind of formula (I)Base quinazoline ditosylate salt
Compound,
Second aspect, the present invention provide pivaloyl amino benzo [d] azepine shown in a kind of formula (I)Base quinazoline ditosylate salt
Close the preparation method of object, the method are as follows: (1) compound shown in formula (II) is mixed with compound shown in formula (III), had
In solvent A, under the action of basic catalyst B, 25~120 DEG C reacted (TLC tracking and monitoring, solvent be acetic acid second
Ester/petroleum ether=1:3 (v/v), preferably 40~100 DEG C 0.5~12h of reaction), after fully reacting, reaction solution is isolated and purified, is made
Obtain compound shown in formula (IV)；The organic solvent A is selected from one of following: chloroform, toluene, methanol, ethyl alcohol, propyl alcohol, isopropanol,
Acetonitrile or N,N-dimethylformamide；The basic catalyst B is selected from one of following: pyridine, diethylamine, triethylamine, quinoline,
N, N- dimethylaniline, 4-dimethylaminopyridine, 4- pyrollidinopyridine or sodium carbonate (preferably pyridine, diethylamine, triethylamine, N,
N- dimethylaniline or 4-dimethylaminopyridine)；
(2) formula (IV) compound represented under reducing agent E effect, has been reacted in organic solvent D at 25~100 DEG C
(TLC tracking and monitoring, solvent are ethyl acetate/petroleum ether=1:1 (v/v), preferably 40~80 DEG C 0.5~12h of reaction) entirely, instead
Liquid is answered to filter, the concentrate after filtrate decompression concentration is dry (preferably 25 DEG C vacuum drying), and formula (V) compound represented is made；
The organic solvent D is one of following: chloroform, toluene, methanol, ethyl alcohol, propyl alcohol, isopropanol, acetonitrile or N, N- dimethyl formyl
Amine；The reducing agent E is one of following: iron powder/concentrated hydrochloric acid, iron powder/acetic acid, palladium carbon/ammonium formate or palladium carbon/hydrazine hydrate；It is described
Iron powder/concentrated hydrochloric acid refers to that the mixing of iron powder and concentrated hydrochloric acid arbitrary proportion, iron powder/acetic acid refer to the mixed of iron powder and acetic acid arbitrary proportion
It closes, the palladium carbon/ammonium formate refers to the mixing of palladium carbon Yu ammonium formate arbitrary proportion, and the palladium carbon/hydrazine hydrate is palladium carbon and hydration
The mixture of hydrazine arbitrary proportion；
(3) compound shown in formula (V) is mixed with pivaloyl chloride or pivalic acid acid anhydride, under basic catalyst F effect, in
In organic solvent G, -10~50 DEG C of fully reactings (TLC tracking and monitoring, solvent are ethyl acetate/petroleum ether=1:1 (v/v),
It is preferred that -10~50 DEG C of 3~12h of reaction), reaction solution is post-treated, and formula (I) compound represented is made；The base catalysis
Agent F is one of following: pyridine, diethylamine, triethylamine, quinoline, N, N- dimethylaniline, 4-dimethylaminopyridine, 4- pyrrolidinyl
Pyridine or sodium carbonate；The organic solvent G is one of following: tetrahydrofuran, methylene chloride, chloroform, ethyl acetate, ether, second
Nitrile, toluene or benzene.
Further, in step (1), compound shown in the formula (III) and compound, basic catalyst B shown in formula (II)
The ratio between the amount of substance of feeding intake is 1.0 ﹕, 0.8~1.2 ﹕ 1.0~8.0.
Further, in step (1), the dosage of the organic solvent A is calculated as 10 with the quality of compound shown in formula (III)~
50mL/g。
Further, the method that reaction solution described in step (1) of the present invention isolates and purifies are as follows: after fully reacting, by reaction solution
Solvent is evaporated off, concentrate is taken to be dissolved with organic solvent C, obtain lysate, then into lysate be added concentrate 1.0~
After mixing, solvent is evaporated off in the column chromatography silica gel of 2.0 times of weight, dry, the mixture of concentrate and silica gel is obtained, by mixture
Fill column, then using volume ratio for 1:0.1~10 petroleum ether and ethyl acetate mixture as eluant, eluent, collect contain target components
Efflux (preferably with ethyl acetate/petroleum ether=1:3 (v/v) be solvent tracing detection, collect target components, preferably receive
Collect the component that Rf value is 0.5), it is concentrated under reduced pressure, dry (preferably 50 DEG C of dryings) obtain formula (IV) compound represented；It is described to have
Solvent C is one of following: ethyl alcohol, chloroform, tetrahydrofuran or ethyl acetate.The organic solvent C dosage is residual can dissolve
Stay object.
Further, in step (2), when the reducing agent E is iron powder/concentrated hydrochloric acid or iron powder/acetic acid, formula (IV) institute
The mass ratio that feeds intake of the iron powder in compound and reducing agent E, concentrated hydrochloric acid or the acetic acid that show is 1.0 ﹕, 1.0~3.0 ﹕ 0.2~1.0.
In the present invention, concentrated hydrochloric acid mass concentration is 36%~38%, and acetic acid uses glacial acetic acid.
Further, in step (2), when the reducing agent E is palladium carbon/ammonium formate or palladium carbon/hydrazine hydrate, formula (IV) institute
The compound that shows and the mass ratio that feeds intake of palladium carbon in reducing agent E, ammonium formate or hydrazine hydrate be 1.0 ﹕, 0.1~0.5 ﹕ 1.0~
3.0.In the present invention be applicable in palladium carbon in palladium mass loading amount be 2~10%, preferably 5%, hydrazine hydrate mass concentration be 40~
80%, preferably 80%.
Further, in step (2), the dosage of the organic solvent D is calculated as 10 with the quality of formula (IV) compound represented
~50mL/g.
Further, in step (3), compound shown in the formula (V) and pivaloyl chloride or pivalic acid acid anhydride, base catalysis
The ratio between amount for the substance that feeds intake of agent F is 1 ﹕, 1.0~8.0 ﹕ 1.0~3.0.
Further, in step (3), the dosage of the organic solvent G is calculated as 11 with the quality of compound shown in formula (V)~
100mL/g。
Further, the specific recommendation step of the present invention (3) carries out as follows: under the conditions of -10~10 DEG C, toward formula
(V) in the organic solvent G solution of compound shown in and basic catalyst F or toward compound and base catalysis shown in formula (V)
The organic solvent G solution of pivaloyl chloride or pivalic acid acid anhydride is added dropwise in agent F, drop finishes, and -10~50 DEG C are reacted 3~12 hours, and gained is anti-
Answer liquid is post-treated to obtain compound shown in formula (I)；The organic solvent volume dosage of pivaloyl chloride or pivalic acid acid anhydride is dissolved to this
Invention does not influence, and total dosage of the organic solvent G is calculated as 11~100mL/g with the quality of compound shown in formula (V).It is organic
Total dosage of solvent G refer to dissolution basic catalyst F and formula (V) shown in compound organic solvent G and dissolution pivaloyl chloride or
The total volume of pivalic acid acid anhydride organic solvent G.
Further, the method for step (3) the of the present invention reaction solution post-processing are as follows: after fully reacting, reaction solution is filtered,
Filtrate steaming removal solvent takes concentrate to be dissolved with organic solvent H, obtains lysate, concentrate is then added into lysate
After mixing, solvent is evaporated off in the column chromatography silica gel of 1.0~2.0 times of weight, dry, is obtained the mixture of concentrate and silica gel, will be mixed
Close object fill column, then using volume ratio for 1:0.1~10 petroleum ether and ethyl acetate mixture as eluant, eluent, collect contain target
The efflux (preferably with ethyl acetate/petroleum ether=1:1 (v/v) for solvent tracing detection, collecting target components) of component, subtracts
Pressure concentration, dry (preferably 50 DEG C of dryings) obtain formula (I) compound represented；The organic solvent H is one of following: ethyl alcohol,
Chloroform, tetrahydrofuran or ethyl acetate.The organic solvent H dosage is with being capable of dissolution residual substance.
Organic solvent A of the present invention, C, D, G and H are organic solvent, organic used in different step for the ease of distinguishing
Solvent is different and names, and letter itself does not have meaning；The catalyst B, reducing agent E and catalyst F are catalyst, in order to just
It is named in differentiation different step used catalyst difference, letter itself does not have meaning.
The third aspect, the invention further relates to pivaloyl amino benzo [d] azepines shown in formula (I)Base quinazoline compounds
Application in preparation prevention or tumor, particularly useful for making answering in prevention or treatment human breast carcinoma drug
With.
Preferably, the drug is with the inhibition active drug of MCF-7 cell strainHJ2mm.Provided by the inventionization
Closing object (I) has preferable inhibitory effect to MCF-7 cell strainHJ2mm.
Pivaloyl amino benzo [d] azepine shown in formula (I) of the present inventionBase quinazoline compounds are also to human lung cancer
Cell strain A-549, people in loop strain HL-60 or human cervical carcinoma cell lines Siha have significant inhibiting effect,
It can be applied in the drug of preparation prevention or treatment human lung cancer, human leukemia or human cervical carcinoma.
The beneficial effects are mainly reflected as follows: (1) of the present invention pivaloyl amino benzo [d] azepineBase quinoline
The anticancer activity that oxazoline compound (I) has had is expected in the drug for being applied to preparation prevention or treatment tumor disease, especially
It is the application in human breast carcinoma, human lung cancer, human leukemia or human cervical carcinoma's drug；(2) pivaloyl aminobenzene provided by the invention
And [d] azepineThe preparation method of base quinazoline compounds (I), simple easily operated, raw material is easy to get, and production cost compared with
It is low, it is suitable for practical.
(4) specific embodiment
The present invention is further described in conjunction with specific embodiments, embodiment below illustrate it is of the invention, rather than
It limit the invention in any way.
Compound (II) prepare reference literature (Weinstock, J.et al.J.Med.Chem., 1986,29 (11),
Method 2315-2325) is prepared.The chloro- 6- nitro-quinazoline (III) of 4- prepares reference literature (Fernandes, C.et
Al.Bioorg.Med.Chem., 2007,15 (12), 3974-3980) method be prepared.
Palladium carbon (Pd/C) model D5H5A that the embodiment of the present invention uses, is purchased from Shaanxi Ruike New Materials Co., Ltd..
Embodiment 1: nitro benzo [d] azepineThe preparation of base quinazoline (IV)
Successively by the chloro- 6- nitro-quinazoline (III) of 1.20 grams of (5.73mmol) 4- and 2.39 grams of (6.87mmol) compounds
(II), 3.62 grams of (45.76mmol) pyridines, 12 milliliters of chloroforms are added in 50 milliliters of reaction flask, are heated to 40 DEG C, TLC tracking
Detection (solvent is ethyl acetate/petroleum ether=1:3 (v/v)), is stirred to react 10 hours, closes reaction, reaction solution is evaporated off molten
10 milliliters of ethyl acetate are added in obtained concentrate and are dissolved, obtains lysate, 3.0 grams of columns is added into lysate for agent
Chromatographic silica gel (300~400 mesh column chromatography silica gel), after mixing, is evaporated off solvent, obtains the mixture of dry concentrate and silica gel,
Mixture is filled into column, then using volume ratio for 1:10 petrol ether/ethyl acetate mixed solution as eluant, eluent, elution, TLC tracking
Detection (solvent is ethyl acetate/petroleum ether=1:3 (v/v)), is collected according to TLC detection containing formula (IV) compound represented
Eluent (Rf value is 0.5), collection liquid concentration, 50 DEG C are dried to obtain faint yellow solid product shown in formula (IV), yield
85.1%, 164~166 DEG C of fusing point.1H NMR(500MHz,CDCl3) 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。
Embodiment 2: nitro benzo [d] azepineThe preparation of base quinazoline (IV)
Successively by the chloro- 6- nitro-quinazoline (III) of 1.20 grams of (5.73mmol) 4- and 1.59 grams of (4.57mmol) compounds
(II), 1.67 grams of (22.83mmol) diethylamine, 60 milliliters of toluene are added in 100 milliliters of three-necked flask, are heated to 100 DEG C,
TLC tracing detection (solvent is ethyl acetate/petroleum ether=1:3 (v/v)), is stirred to react 2 hours, closes reaction, reaction solution
Solvent is evaporated off, 20 milliliters of ethyl alcohol are added in obtained concentrate and are dissolved, lysate is obtained, 2.5 grams are added into lysate
Column chromatography silica gel (300~400 mesh column chromatography silica gel), after mixing, is evaporated off solvent, obtains the mixing of dry concentrate and silica gel
Mixture is filled column by object, then using volume ratio for 1:5 petrol ether/ethyl acetate mixed solution as eluant, eluent, elution, TLC with
Track detection (solvent is ethyl acetate/petroleum ether=1:3 (v/v)), collects according to TLC detection and contains formula (IV) compound represented
Eluent (Rf value be 0.5), collection liquid concentration, 50 DEG C are dried to obtain faint yellow solid product shown in formula (IV), yield
72.6%, 164~166 DEG C of fusing point.1H NMR and IR is the same as embodiment 1.
Embodiment 3: nitro benzo [d] azepineThe preparation of base quinazoline (IV)
Successively by the chloro- 6- nitro-quinazoline (III) of 1.20 grams of (5.73mmol) 4- and 1.99 grams of (5.72mmol) compounds
(II), 0.58 gram of (5.73mmol) triethylamine, 60 milliliters of ethyl alcohol are added in 100 milliliters of three-necked flask, are heated to 60 DEG C, TLC
Tracing detection (solvent is ethyl acetate/petroleum ether=1:3 (v/v)), is stirred to react 8 hours, closes reaction, reaction solution is evaporated off
20 milliliters of chloroforms are added in obtained concentrate and are dissolved, obtains lysate, 2.5 grams of column layers is added into lysate for solvent
It analyses silica gel (300~400 mesh column chromatography silica gel), after mixing, solvent is evaporated off, obtains the mixture of dry concentrate and silica gel, it will
Mixture fill column, then using volume ratio for 10:1 petrol ether/ethyl acetate mixed solution as eluant, eluent, elution, TLC tracking inspection
(solvent is ethyl acetate/petroleum ether=1:3 (v/v)) is surveyed, is detected according to TLC and collects washing for (IV) compound represented Han formula
De- liquid (Rf value is 0.5), collection liquid concentration, 50 DEG C are dried to obtain faint yellow solid product shown in formula (IV), yield 77.2%,
164~166 DEG C of fusing point.1H NMR and IR is the same as embodiment 1.
Embodiment 4: nitro benzo [d] azepineThe preparation of base quinazoline (IV)
Successively by the chloro- 6- nitro-quinazoline (III) of 1.20 grams of (5.73mmol) 4- and 2.20 grams of (6.32mmol) compounds
(II), 1.40 grams of (11.46mmol) 4-dimethylaminopyridine, 60 milliliters of isopropanols are added in 100 milliliters of three-necked flask, room temperature
25 DEG C of stirrings, TLC tracing detection (solvent is ethyl acetate/petroleum ether=1:3 (v/v)), react 12 hours, close reaction,
Solvent is evaporated off in reaction solution, and 20 milliliters of tetrahydrofurans are added in obtained concentrate and are dissolved, lysate are obtained, into lysate
4.0 grams of column chromatography silica gels (300~400 mesh column chromatography silica gel) is added, after mixing, solvent is evaporated off, obtains dry concentrate and silicon
Mixture is filled column by the mixture of glue, then using volume ratio for 5:1 petrol ether/ethyl acetate mixed solution as eluant, eluent, wash
De-, TLC tracing detection (solvent is ethyl acetate/petroleum ether=1:3 (v/v)) is collected according to TLC detection containing shown in formula (IV)
Compound eluent (Rf value be 0.5), collection liquid concentration, 50 DEG C are dried to obtain the production of faint yellow solid shown in formula (IV)
Object, yield 80.2%, 164~166 DEG C of fusing point.1H NMR and IR is the same as embodiment 1.
Embodiment 5: nitro benzo [d] azepineThe preparation of base quinazoline (IV)
Successively by the chloro- 6- nitro-quinazoline (III) of 1.20 grams of (5.73mmol) 4- and 1.79 grams of (5.15mmol) compounds
(II), 1.04 grams of (8.58mmol) N, N- dimethylanilines, 12 milliliters of n,N-Dimethylformamide are added in 50 milliliters of reaction flask,
120 DEG C are heated to, TLC tracing detection (solvent is ethyl acetate/petroleum ether=1:3 (v/v)) is stirred to react 0.5 hour, closes
Reaction is closed, solvent is evaporated off in reaction solution, and 20 milliliters of tetrahydrofurans are added in obtained concentrate and are dissolved, lysate is obtained, to
5.0 grams of column chromatography silica gels (300~400 mesh column chromatography silica gel) are added in lysate, after mixing, solvent is evaporated off, obtain dry dense
Mixture is filled column by the mixture of contracting object and silica gel, is then for the petrol ether/ethyl acetate mixed solution of 1:1 with volume ratio
Eluant, eluent, elution, TLC tracing detection (solvent is ethyl acetate/petroleum ether=1:3 (v/v)) are collected according to TLC detection and are contained
The eluent (Rf value be 0.5) of formula (IV) compound represented, collection liquid concentration, 50 DEG C be dried to obtain it is yellowish shown in formula (IV)
Color solid product, yield 89.6%, 164~166 DEG C of fusing point.1HNMR and IR is the same as embodiment 1.
Embodiment 6: nitro benzo [d] azepineThe preparation of base quinazoline (IV)
Successively by the chloro- 6- nitro-quinazoline (III) of 1.20 grams of (5.73mmol) 4- and 2.39 grams of (6.87mmol) compounds
(II), 3.62 grams of (45.76mmol) pyridines, 20 milliliters of propyl alcohol are added in 50 milliliters of reaction flask, are heated to 40 DEG C, TLC tracking
Detection (solvent is ethyl acetate/petroleum ether=1:3 (v/v)), is stirred to react 10 hours, closes reaction, reaction solution is evaporated off molten
20 milliliters of ethyl acetate are added in obtained concentrate and are dissolved, obtains lysate, 3.5 grams of columns is added into lysate for agent
Chromatographic silica gel (300~400 mesh column chromatography silica gel), after mixing, is evaporated off solvent, obtains the mixture of dry concentrate and silica gel,
Mixture is filled into column, then using volume ratio for 1:1 petrol ether/ethyl acetate mixed solution as eluant, eluent, elution, TLC tracking
Detection (solvent is ethyl acetate/petroleum ether=1:3 (v/v)), is collected according to TLC detection containing formula (IV) compound represented
Eluent (Rf value is 0.5), collection liquid concentration, 50 DEG C are dried to obtain faint yellow solid product shown in formula (IV), yield
78.3%, 164~166 DEG C of fusing point.1H NMR and IR is the same as embodiment 1.
Embodiment 7: amino benzo [d] azepineThe preparation of base quinazoline (V)
0.40 gram of (0.77mmol) nitro benzo [d] azepine successively prepared by 1 method of embodimentBase quinazoline (IV),
0.40 gram of (6.34mmol) ammonium formate, 0.04 gram of 5%Pd/C, 4.0 milliliters of chloroforms are added in reaction flask, 25 DEG C of room temperature stirrings,
TLC tracing detection (solvent is ethyl acetate/petroleum ether=1:1 (v/v)), reacts 12 hours, filtering, filtrate concentration, 25 DEG C
Vacuum drying obtains faint yellow solid product amino benzo [d] azepineBase quinazoline (V), yield 98.2%, fusing point 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。
Embodiment 8: amino benzo [d] azepineThe preparation of base quinazoline (V)
0.40 gram of (0.77mmol) nitro benzo [d] azepine successively prepared by 2 method of embodimentBase quinazoline (IV),
1.20 grams of (19.18mmol) 80wt% hydrazine hydrates, 0.20 gram of 5%Pd/C, 20.0 milliliters of toluene are added to 50 milliliters of reaction flask
In, 100 DEG C are heated to, it is small to be stirred to react 0.5 for TLC tracing detection (solvent is ethyl acetate/petroleum ether=1:1 (v/v))
When, cold filtration, filtrate is concentrated, and 25 DEG C of vacuum drying obtain faint yellow solid product amino benzo [d] azepineBase quinazoline
(V), yield 100.0%, 122~126 DEG C of fusing point.1H NMR and IR is the same as embodiment 7.
Embodiment 9: amino benzo [d] azepineThe preparation of base quinazoline (V)
0.40 gram of (0.77mmol) nitro benzo [d] azepine successively prepared by 3 method of embodimentBase quinazoline (IV),
0.08 gram of concentrated hydrochloric acid (mass concentration 36~38%), 0.40 gram of iron powder, 20.0 ml methanols are added in 50 milliliters of reaction flask,
40 DEG C are heated to, TLC tracing detection (solvent is ethyl acetate/petroleum ether=1:1 (v/v)) is stirred to react 8 hours, cooling
Filtering, filtrate concentration, 25 DEG C of vacuum drying obtain faint yellow solid product amino benzo [d] azepineBase quinazoline (V) is received
Rate 94.1%, 122~126 DEG C of fusing point.1HNMR and IR is the same as embodiment 7.
Embodiment 10: amino benzo [d] azepineThe preparation of base quinazoline (V)
0.40 gram of (0.77mmol) nitro benzo [d] azepine successively prepared by 4 method of embodimentBase quinazoline (IV),
0.40 gram of acetic acid, 1.20 grams of iron powders, 20.0 milliliters of isopropanols are added in 50 milliliters of reaction flask, are heated to 80 DEG C, TLC tracking
Detection (solvent is ethyl acetate/petroleum ether=1:1 (v/v)), is stirred to react 3 hours, cold filtration, filtrate concentration, 25 DEG C
Vacuum drying obtains faint yellow solid product amino benzo [d] azepineBase quinazoline (V), yield 97.5%, fusing point 122~
126℃。1H NMR and IR is the same as embodiment 7.
Embodiment 11: pivaloyl amino benzo [d] azepineThe preparation of base quinazoline (I)
0.27 gram of (0.55mmol) amino benzo [d] azepine successively prepared by 7 method of embodimentBase quinazoline (V),
0.13 gram of (1.64mmol) pyridine, 3 milliliters of tetrahydrofurans are added in reaction flask, and 0.531 gram is added dropwise under -10 DEG C of stirring conditions
(4.40mmol) pivaloyl chloride, drop finish, TLC tracing detection (solvent is ethyl acetate/petroleum ether=1:1), under the conditions of -10 DEG C
Reaction 12 hours, filtering, filtrate steaming removal solvent, concentrate are added 10 milliliters of ethyl acetate and are dissolved, and obtain lysate, Xiang Rong
It solves and 0.60 gram of column chromatography silica gel (300~400 mesh column chromatography silica gel) is added in liquid, after mixing, solvent is evaporated off, obtains dry concentration
Mixture is filled column by the mixture of object and silica gel, and then the petrol ether/ethyl acetate mixed solution with volume ratio for 1:10 is to wash
De- agent, elution, TLC tracing detection (solvent is ethyl acetate/petroleum ether=1:1 (v/v)) are collected according to TLC detection and contain formula
(I) eluent (Rf value is 0.5) of compound represented, collection liquid concentration, 50 DEG C are dried to obtain pivaloyl ammonia shown in formula (I)
Base benzo [d] azepineBase quinazoline pale solid, yield 64.2%, 131~133 DEG C of fusing point.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。
Embodiment 12: pivaloyl amino benzo [d] azepineThe preparation of base quinazoline (I)
0.27 gram of (0.55mmol) amino benzo [d] azepine successively prepared by 8 method of embodimentBase quinazoline (V),
0.04 gram of (0.55mmol) diethylamine, 10.0 milliliters of chloroforms are added in 50 milliliters of reaction flask, are added dropwise under 10 DEG C of stirring conditions
0.066 gram of (0.55mmol) pivaloyl chloride and 5.0 milliliters of chloroform mixed solutions, drop finish, and (solvent is acetic acid to TLC tracing detection
Ethyl ester/petroleum ether=1:1 (v/v)), it reacts 8 hours under the conditions of 10 DEG C, filters, filtrate steaming removal solvent, concentrate is added 20 milliliters
Ethyl alcohol is dissolved, and lysate is obtained, and 0.26 gram of column chromatography silica gel (300~400 mesh column chromatography silica gel) is added into lysate,
After mixing, solvent is evaporated off, obtains the mixture of dry concentrate and silica gel, mixture is filled into column, is then 1:5's with volume ratio
Petrol ether/ethyl acetate mixed solution is eluant, eluent, and elution, (solvent is ethyl acetate/petroleum ether=1:1 to TLC tracing detection
(v/v)) eluent (Rf value is 0.5) containing formula (I) compound represented, is collected according to TLC detection, collection liquid is concentrated, and 50 DEG C
It is dried to obtain pivaloyl amino benzo [d] azepine shown in formula (I)Base quinazoline pale solid, yield 74.7%, fusing point
131~133 DEG C.1H NMR and IR is same
Embodiment 11.
Embodiment 13: pivaloyl amino benzo [d] azepineThe preparation of base quinazoline (I)
0.27 gram of (0.55mmol) amino benzo [d] azepine successively prepared by 9 method of embodimentBase quinazoline (V),
0.111 gram of (1.10mmol) triethylamine, 10.0 milliliters of ethyl acetate are added in 50 milliliters of reaction flask, under 0 DEG C of stirring condition
0.133 gram of (1.10mmol) pivaloyl chloride and 5.0 milliliters of ethyl acetate solutions are added dropwise, drop finishes, and (solvent is TLC tracing detection
Ethyl acetate/petroleum ether=1:1), it reacts 6 hours under the conditions of 25 DEG C, filters, filtrate steaming removal solvent, concentrate is added 20 milliliters
Chloroform is dissolved, and lysate is obtained, and 0.30 gram of column chromatography silica gel (300~400 mesh column chromatography silica gel) is added into lysate,
After mixing, solvent is evaporated off, obtains the mixture of dry concentrate and silica gel, mixture is filled into column, is then 10:1 with volume ratio
Petrol ether/ethyl acetate mixed solution be eluant, eluent, elution, TLC tracing detection (solvent be ethyl acetate/petroleum ether=
1:1 (v/v)), the eluent (Rf value is 0.5) containing formula (I) compound represented is collected according to TLC detection, collection liquid is concentrated, and 50
DEG C it is dried to obtain pivaloyl amino benzo [d] azepine shown in formula (I)Base quinazoline pale solid, yield 50.5% melt
131~133 DEG C of point.1H NMR and IR is the same as embodiment 11.
Embodiment 14: pivaloyl amino benzo [d] azepineThe preparation of base quinazoline (I)
0.27 gram of (0.55mmol) amino benzo [d] azepine successively prepared by 10 method of embodimentBase quinazoline
(V), 0.067 gram of (0.55mmol) 4-dimethylaminopyridine, 20.0 milliliters of toluene are added in 50 milliliters of reaction flask, and 5 DEG C are stirred
The solution of 0.410 gram of (2.20mmol) pivalic acid acid anhydride and 7.0 milliliters of toluene is added dropwise under the conditions of mixing, drop finishes, and is heated to 50 DEG C, TLC
Tracing detection (solvent is ethyl acetate/petroleum ether=1:1), reacts 3 hours, filtering, filtrate steaming removal solvent, concentrate is added
20 milliliters of tetrahydrofurans are dissolved, and lysate is obtained, and 0.40 gram of column chromatography silica gel (300~400 mesh column is added into lysate
Chromatographic silica gel), after mixing, solvent is evaporated off, obtains the mixture of dry concentrate and silica gel, mixture is filled into column, then with body
Product than be 5:1 petrol ether/ethyl acetate mixed solution be eluant, eluent, elution, TLC tracing detection (solvent for ethyl acetate/
Petroleum ether=1:1 (v/v)), the eluent (Rf value is 0.5) containing formula (I) compound represented is collected according to TLC detection, is collected
Liquid concentration, 50 DEG C are dried to obtain pivaloyl amino benzo [d] azepine shown in formula (I)Base quinazoline pale solid, yield
57.4% 131~133 DEG C of fusing point.1H NMR and IR is the same as embodiment 11.
Embodiment 15: pivaloyl amino benzo [d] azepineThe preparation of base quinazoline (I)
0.27 gram of (0.55mmol) amino benzo [d] azepine successively prepared by 7 method of embodimentBase quinazoline (V),
0.213 gram of (1.65mmol) quinoline, 15.0 milliliters of benzene are added in 50 milliliters of reaction flask, are added dropwise under -10 DEG C of stirring conditions
The solution of 0.265 gram of (2.20mmol) pivaloyl chloride and 5.0 milliliters of benzene, drop finish, TLC tracing detection (solvent be ethyl acetate/
Petroleum ether=1:1), it reacts 12 hours under the conditions of -10 DEG C, filters, filtrate steaming removal solvent, 20 milliliters of tetrahydrofurans are added in concentrate
It is dissolved, obtains lysate, 0.40 gram of column chromatography silica gel (300~400 mesh column chromatography silica gel) is added into lysate, mixed
Afterwards, solvent is evaporated off, obtains the mixture of dry concentrate and silica gel, mixture is filled into column, is then the petroleum of 1:1 with volume ratio
Ether/ethyl acetate mixture is eluant, eluent, and elution, (solvent is ethyl acetate/petroleum ether=1:1 (v/ to TLC tracing detection
V)), the eluent (Rf value is 0.5) containing formula (I) compound represented, collection liquid concentration, 50 DEG C of dryings are collected according to TLC detection
Obtain pivaloyl amino benzo [d] azepine shown in formula (I)Base quinazoline pale solid, yield 70.9%, fusing point 131~
133℃。1H NMR and IR is the same as embodiment 11.
Embodiment 16: pivaloyl amino benzo [d] azepineThe preparation of base quinazoline (I)
0.27 gram of (0.55mmol) amino benzo [d] azepine successively prepared by 7 method of embodimentBase quinazoline (V),
0.164 gram of (1.10mmol) 4- pyrollidinopyridine, 15.0 milliliters of methylene chloride are added in 50 milliliters of reaction flask, and 10 DEG C are stirred
0.133 gram of (1.10mmol) pivaloyl chloride and 5.0 milliliters of dichloromethane solutions are added dropwise under the conditions of mixing, drop finishes, TLC tracing detection
(solvent is ethyl acetate/petroleum ether=1:1), is reacted 8 hours under the conditions of 10 DEG C, is filtered, and filtrate steaming removal solvent, concentrate adds
Enter 20 milliliters of ethyl alcohol to be dissolved, obtain lysate, 0.50 gram of column chromatography silica gel (300~400 mesh column layer is added into lysate
Analyse silica gel), after mixing, solvent is evaporated off, obtains the mixture of dry concentrate and silica gel, mixture is filled into column, then with volume
Than being eluant, eluent for the petrol ether/ethyl acetate mixed solution of 10:1, elution, TLC tracing detection (solvent be ethyl acetate/
Petroleum ether=1:1 (v/v)), the eluent (Rf value is 0.5) containing formula (I) compound represented is collected according to TLC detection, is collected
Liquid concentration, 50 DEG C are dried to obtain pivaloyl amino benzo [d] azepine shown in formula (I)Base quinazoline pale solid, yield
68.6%, 131~133 DEG C of fusing point.1H NMR and IR is the same as embodiment 11.
Embodiment 17: anticancer activity testing in vitro
(1) compound obtained (I), (IV) and (V) MCF-7 cell strainHJ2mm biological activity test has been subjected to.
Test method: tetrazolium reduction method (mtt assay).
Cell strain: MCF-7 cell strainHJ2mm.Above-mentioned tumor cell line is purchased from Chinese Academy of Sciences Shanghai school of life and health sciences
Cell bank.
Experimental procedure is as follows:
(a) preparation of sample: for solvable sample, every 1mg is dissolved with 40 μ L DMSO, takes 2 μ L dilute with 1000 μ L culture mediums
It releases, makes 100 μ g/mL of concentration, then with culture solution serial dilution to using concentration.
(b) culture of cell
1. the preparation of culture medium: containing 800,000 units of Penicillin, 1.0g strepto- in every 1000mL DMEM culture medium (Gibco)
Element, 10% inactivated fetal bovine serum.
2. the culture of cell: by tumor cell inoculation in culture medium, setting 37 DEG C, 5%CO2It is cultivated in incubator, 3~5d
Passage.
3. measuring sample to the inhibiting effect of growth of tumour cell
10th generation cell EDTA- pancreatin digestive juice is digested, and is diluted to 1 × 10 with culture medium6/ mL is added to 96 holes
In tissue culture plate, every 100 μ L of hole sets 37 DEG C, 5%CO2It is cultivated in incubator.After inoculation for 24 hours, it is separately added into dilute with culture medium
100 μ g/mL, the 10 μ g/mL and 1 μ g/mL sample released, every 100 μ L of hole, each concentration add 3 holes, set 37 DEG C, 5%CO2Incubator
MTT, the every 10 μ L of hole of 5mg/mL are added after 72h in cell culture well for middle culture, set 37 DEG C of incubations 3h, addition DMSO, every hole
150 μ L, are vibrated with oscillator, and Shi formazan is completely dissolved, with microplate reader under 570nm wavelength colorimetric.To be free of under similarity condition
Sample, culture medium culture containing same concentration DMSO cell as control, calculate sample to the IC of growth of tumour cell50。
The results are shown in Table 1 for test:
The inhibiting effect that 1. compound of table (I), (IV) and (V) grows cancer cell line MCF-7
(2) according to embodiment 11, pivaloyl chloride is used to 4- iodobenzoyl chloride, 3- methoxy benzoyl chloride or cinnamoyl respectively
Chloro replaces, other operations have been respectively synthesized quinazoline compounds (a) with embodiment 11, and (b) and (c), structure are as follows:
According to the above method by quinazoline compounds (a) obtained, (b) and (c) has carried out Breast cancer lines
MCF-7 biological activity test, test result show quinazoline compounds (a), and (b) and (c) is to MCF-7 cell strainHJ2mm
Inhibitory effect is unobvious, compound (a), and (b) and (c) can not show a candle to chemical combination to the anticancer activity of MCF-7 cell strainHJ2mm
Object (I).Concrete outcome is as shown in table 2:
The inhibiting effect that 2. compound (a) of table, (b) and (c) grow cancer cell line MCF-7
Above-mentioned anticancer activity testing in vitro experiment shows: the similar compound (a) of other 3 structures, (b) and (c) is to people
The inhibiting effect of breast cancer cell line mcf-7 growth is unobvious.Compound (I) grows MCF-7 cell strainHJ2mm
Inhibiting effect is significant, hence it is evident that is better than compound (a), (b) and (c).
(3) method of reference literature (Rao, G.-W.et al.ChemMedChem, 2013,8 (6), 928-933) is prepared into
To 4- chloro-quinazoline, further according to embodiment 1, the chloro- 6- nitro-quinazoline of 4- is replaced with 4- chloro-quinazoline, other operations are the same as implementation
Example 1 has synthesized quinazoline compounds (d), and structure is as follows:
Quinazoline compounds (d) obtained MCF-7 cell strainHJ2mm biology has been carried out according to the above method to live
Property test, test result shows that quinazoline compounds (d) can not show a candle to chemical combination to the anticancer activity of MCF-7 cell strainHJ2mm
Object (I).Concrete outcome is as shown in table 3:
The inhibiting effect that 3. compound (d) of table grows cancer cell line MCF-7
(4) according to embodiment 11, pivaloyl chloride is used to chlorobenzoyl chloride, butyl chloride, propionyl chloride, chloracetyl chloride or isobutyl respectively
Acyl chlorides replaces, other operations have been respectively synthesized quinazoline compounds (e), (f), (g), (h) and (j), structure with embodiment 11
It is as follows:
Quinazoline compounds (e) obtained, (f), (g), (h) and (j) human breast carcinoma has been subjected to according to the above method
Cell strain MCF-7 biological activity test, test result show quinazoline compounds (e), (f), (g), (h) with (j) to human milk
The anticancer activity of adenocarcinoma cell strain MCF-7 is not so good as compound (I).Concrete outcome is as shown in table 4:
The inhibiting effect that 4. compound (e) of table, (f), (g), (h) and (j) grow cancer cell line MCF-7
Embodiment 18: anticancer activity testing in vitro
(1) compound obtained (I) and (IV) human lung cancer cell lines A-549 biological activity test has been subjected to.
Test method: tetrazolium reduction method (mtt assay).
Cell strain: human lung cancer cell lines A-549.Above-mentioned tumor cell line is thin purchased from Chinese Academy of Sciences Shanghai school of life and health sciences
Born of the same parents library.
Experimental procedure is as follows:
(a) preparation of sample: for solvable sample, every 1mg is dissolved with 40 μ L DMSO, takes 2 μ L dilute with 1000 μ L culture mediums
It releases, makes 100 μ g/mL of concentration, then with culture solution serial dilution to using concentration.
(b) culture of cell
1. the preparation of culture medium: containing 800,000 units of Penicillin, 1.0g strepto- in every 1000mL DMEM culture medium (Gibco)
Element, 10% inactivated fetal bovine serum.
2. the culture of cell: by tumor cell inoculation in culture medium, setting 37 DEG C, 5%CO2It is cultivated in incubator, 3~5d
Passage.
3. measuring sample to the inhibiting effect of growth of tumour cell
2nd generation cell EDTA- pancreatin digestive juice is digested, and is diluted to 1 × 10 with culture medium6It is thin to be added to 96 holes by/mL
In born of the same parents' culture plate, every 100 μ L of hole sets 37 DEG C, 5%CO2It is cultivated in incubator.After inoculation for 24 hours, it is separately added into and is diluted with culture medium
100 μ g/mL, 10 μ g/mL and 1 μ g/mL sample, every 100 μ L of hole, each concentration adds 3 holes, sets 37 DEG C, 5%CO2In incubator
It cultivates, MTT, the every 10 μ L of hole of 5mg/mL is added after 72h in cell culture well, set 37 DEG C of incubation 3h, DMSO, every hole is added
150 μ L, are vibrated with oscillator, and Shi formazan is completely dissolved, with microplate reader under 570nm wavelength colorimetric.To be free of under similarity condition
Sample, the cell of the culture medium culture containing same concentration DMSO calculate sample to the IC of growth of tumour cell as control50。
The results are shown in Table 5 for test:
The inhibiting effect that 5. compound of table (I) and (IV) grow cancer cell line A-549
(2) according to embodiment 11, pivaloyl chloride is used to 4- iodobenzoyl chloride, 3- methoxy benzoyl chloride or cinnamoyl respectively
Chloro replaces, other operations have been respectively synthesized quinazoline compounds (a) with embodiment 11, and (b) and (c), structure are as follows:
According to the above method by quinazoline compounds (a) obtained, (b) and (c) has carried out human lung cancer cell lines A-549
Biological activity test, test result show quinazoline compounds (a), and (b) and (c) inhibits to imitate to human lung cancer cell lines A-549
Fruit is unobvious, compound (a), and (b) and (c) can not show a candle to compound (I) to the anticancer activity of human lung cancer cell lines A-549.Tool
The results are shown in Table 6 for body:
The inhibiting effect that 6. compound (a) of table, (b) and (c) grow cancer cell line A-549
Above-mentioned anticancer activity testing in vitro experiment shows: the similar compound (a) of other 3 structures, (b) and (c) is to people
The inhibiting effect of lung cancer cell line A-549 growth is unobvious.The inhibition that compound (I) grows human lung cancer cell lines A-549
Effect is significant, hence it is evident that is better than compound (a), (b) and (c).
(3) method of reference literature (Rao, G.-W.et al.ChemMedChem, 2013,8 (6), 928-933) is prepared into
To 4- chloro-quinazoline, further according to embodiment 1, the chloro- 6- nitro-quinazoline of 4- is replaced with 4- chloro-quinazoline, other operations are the same as implementation
Example 1 has synthesized quinazoline compounds (d), and structure is as follows:
Quinazoline compounds (d) obtained human lung cancer cell lines A-549 bioactivity has been subjected to according to the above method
Test, test result show that quinazoline compounds (d) can not show a candle to compound to the anticancer activity of human lung cancer cell lines A-549
(Ⅰ).Concrete outcome is as shown in table 7:
The inhibiting effect that 7. compound (d) of table grows cancer cell line A-549
(4) according to embodiment 11, pivaloyl chloride is replaced with chlorobenzoyl chloride, chloracetyl chloride or isobutyryl chloride respectively, other
Operation has been respectively synthesized quinazoline compounds (e), (h) and (j) with embodiment 11, and structure is as follows:
Quinazoline compounds (e) obtained, (h) and (j) human lung cancer cell lines A-549 has been subjected to according to the above method
Biological activity test, test result show the anticancer of quinazoline compounds (e), (h) with (j) to human lung cancer cell lines A-549
Activity is not so good as compound (I).Concrete outcome is as shown in table 8:
The inhibiting effect that 8. compound (e) of table, (h) and (j) grow cancer cell line A-549
Embodiment 19: anticancer activity testing in vitro
(1) compound obtained (I) people in loop strain HL-60 biological activity test has been subjected to.
Test method: tetrazolium reduction method (mtt assay).
Cell strain: people in loop strain HL-60.Above-mentioned tumor cell line is purchased from Chinese Academy of Sciences Shanghai life
Academy of sciences's cell bank.
Experimental procedure is as follows:
(a) preparation of sample: for solvable sample, every 1mg is dissolved with 40 μ L DMSO, takes 2 μ L dilute with 1000 μ L culture mediums
It releases, makes 100 μ g/mL of concentration, then with culture solution serial dilution to using concentration.
(b) culture of cell
1. the preparation of culture medium: containing 800,000 units of Penicillin, 1.0g strepto- in every 1000mL DMEM culture medium (Gibco)
Element, 10% inactivated fetal bovine serum.
2. the culture of cell: by tumor cell inoculation in culture medium, setting 37 DEG C, 5%CO2It is cultivated in incubator, 3~5d
Passage.
3. measuring sample to the inhibiting effect of growth of tumour cell
2nd generation cell EDTA- pancreatin digestive juice is digested, and is diluted to 1 × 10 with culture medium6It is thin to be added to 96 holes by/mL
In born of the same parents' culture plate, every 100 μ L of hole sets 37 DEG C, 5%CO2It is cultivated in incubator.After inoculation for 24 hours, it is separately added into and is diluted with culture medium
100 μ g/mL, 10 μ g/mL and 1 μ g/mL sample, every 100 μ L of hole, each concentration adds 3 holes, sets 37 DEG C, 5%CO2In incubator
It cultivates, MTT, the every 10 μ L of hole of 5mg/mL is added after 72h in cell culture well, set 37 DEG C of incubation 3h, DMSO, every hole is added
150 μ L, are vibrated with oscillator, and Shi formazan is completely dissolved, with microplate reader under 570nm wavelength colorimetric.To be free of under similarity condition
Sample, the cell of the culture medium culture containing same concentration DMSO calculate sample to the IC of growth of tumour cell as control50。
The results are shown in Table 9 for test:
The inhibiting effect that 9. compound of table (I) grows cancer cell line HL-60
(2) according to embodiment 11, pivaloyl chloride is replaced with 3- methoxy benzoyl chloride or cinnamoyl chloride respectively, other behaviour
Make to be respectively synthesized quinazoline compounds (b) and (c), structure is as follows with embodiment 11:
Quinazoline compounds (b) obtained and (c) people in loop strain has been subjected to according to the above method
HL-60 biological activity test, test result show quinazoline compounds (b) and (c) to people in loop strain HL-
60 inhibitory effects are unobvious, and compound (b) and (c) can not show a candle to the anticancer activity of people in loop strain HL-60
Compound (I).Concrete outcome is as shown in table 10:
The inhibiting effect that 10. compound (b) of table and (c) grow cancer cell line HL-60
Above-mentioned anticancer activity testing in vitro experiment shows: the similar compound (b) of other 2 structures and (c) are to the early children of people
The inhibiting effect of grain leukemia cell line HL-60 growth is unobvious.Compound (I) is to people in loop strain HL-
The inhibiting effect of 60 growths is significant, hence it is evident that better than compound (b) and (c).
(3) according to embodiment 11, pivaloyl chloride is replaced with cyclohexyl methyl chloro-formate or propionyl chloride respectively, other behaviour
Make to be respectively synthesized quinazoline compounds (k) and (g), structure is as follows with embodiment 11:
Quinazoline compounds (k) obtained and (g) people in loop strain has been subjected to according to the above method
HL-60 biological activity test, test result show quinazoline compounds (k) and (g) to people in loop strain HL-
60 anticancer activity is not so good as compound (I).Concrete outcome is as shown in table 11:
The inhibiting effect that 11. compound (k) of table and (g) grow cancer cell line HL-60
Embodiment 20: anticancer activity testing in vitro
(1) compound obtained (I) human cervical carcinoma cell lines Siha biological activity test has been subjected to.
Test method: tetrazolium reduction method (mtt assay).
Cell strain: human cervical carcinoma cell lines Siha.Above-mentioned tumor cell line is purchased from Chinese Academy of Sciences Shanghai school of life and health sciences
Cell bank.
Experimental procedure is as follows:
(a) preparation of sample: for solvable sample, every 1mg is dissolved with 40 μ L DMSO, takes 2 μ L dilute with 1000 μ L culture mediums
It releases, makes 100 μ g/mL of concentration, then with culture solution serial dilution to using concentration.
(b) culture of cell
1. the preparation of culture medium: containing 800,000 units of Penicillin, 1.0g strepto- in every 1000mL DMEM culture medium (Gibco)
Element, 10% inactivated fetal bovine serum.
2. the culture of cell: by tumor cell inoculation in culture medium, setting 37 DEG C, 5%CO2It is cultivated in incubator, 3~5d
Passage.
3. measuring sample to the inhibiting effect of growth of tumour cell
2nd generation cell EDTA- pancreatin digestive juice is digested, and is diluted to 1 × 10 with culture medium6It is thin to be added to 96 holes by/mL
In born of the same parents' culture plate, every 100 μ L of hole sets 37 DEG C, 5%CO2It is cultivated in incubator.After inoculation for 24 hours, it is separately added into and is diluted with culture medium
100 μ g/mL, 10 μ g/mL and 1 μ g/mL sample, every 100 μ L of hole, each concentration adds 3 holes, sets 37 DEG C, 5%CO2In incubator
It cultivates, MTT, the every 10 μ L of hole of 5mg/mL is added after 72h in cell culture well, set 37 DEG C of incubation 3h, DMSO, every hole is added
150 μ L, are vibrated with oscillator, and Shi formazan is completely dissolved, with microplate reader under 570nm wavelength colorimetric.To be free of under similarity condition
Sample, the cell of the culture medium culture containing same concentration DMSO calculate sample to the IC of growth of tumour cell as control50.It surveys
The result of examination is as shown in table 12:
The inhibiting effect that 12. compound of table (I) grows cancer cell line Siha
(2) according to embodiment 11, pivaloyl chloride is replaced with 3- methoxy benzoyl chloride or cinnamoyl chloride respectively, other behaviour
Make to be respectively synthesized quinazoline compounds (b) and (c), structure is as follows with embodiment 11:
Quinazoline compounds (b) obtained and (c) human cervical carcinoma cell lines Siha life has been subjected to according to the above method
Object active testing, test result show quinazoline compounds (b) and (c) to human cervical carcinoma cell lines Siha inhibitory effect not
Obviously, compound (b) and (c) can not show a candle to compound (I) to the anticancer activity of human cervical carcinoma cell lines Siha.Concrete outcome such as table
Shown in 13:
The inhibiting effect that 13. compound (b) of table and (c) grow cancer cell line Siha
Above-mentioned anticancer activity testing in vitro experiment shows: the similar compound (b) of other 2 structures and (c) are to people's uterine neck
The inhibiting effect of cancer cell line Siha growth is unobvious.Compound (I) makees the inhibition that human cervical carcinoma cell lines Siha is grown
With significant, hence it is evident that better than compound (b) and (c).
(3) according to embodiment 11, pivaloyl chloride is replaced with butyl chloride or cyclohexyl methyl chloro-formate respectively, other behaviour
Make to be respectively synthesized quinazoline compounds (f) and (k), structure is as follows with embodiment 11:
Quinazoline compounds (f) obtained and (k) human cervical carcinoma cell lines Siha life has been subjected to according to the above method
Object active testing, test result show that compound (f) and (k) can not show a candle to chemical combination to the anticancer activity of human cervical carcinoma cell lines Siha
Object (I).Concrete outcome is as shown in table 14:
The inhibiting effect that 14. compound (f) of table and (k) grow cancer cell line Siha
Claims (10)
1. pivaloyl amino benzo [d] azepine shown in a kind of formula (I)Base quinazoline compounds:
2. pivaloyl amino benzo [d] azepine shown in a kind of formula as described in claim 1 (I)Base quinazoline compounds
Preparation method, it is characterised in that the method are as follows:
(1) compound shown in formula (II) is mixed with compound shown in formula (III), in organic solvent A, in basic catalyst B's
Under effect, 25~120 DEG C are reacted, and after fully reacting, reaction solution is isolated and purified, and compound shown in formula (IV) is made；Institute
Organic solvent A is stated selected from one of following: chloroform, toluene, methanol, ethyl alcohol, propyl alcohol, isopropanol, acetonitrile or N, N- dimethyl formyl
Amine；The basic catalyst B is selected from one of following: pyridine, diethylamine, triethylamine, quinoline, N, N- dimethylaniline, 4- diformazan
Aminopyridine, 4- pyrollidinopyridine or sodium carbonate；
(2) formula (IV) compound represented obtained by step (1) is dissolved in organic solvent D, under reducing agent E effect, 25
~100 DEG C of fully reactings, reaction solution filtering, the concentrate after filtrate decompression concentration is dry, and formula (V) compound represented is made；
The organic solvent D is one of following: chloroform, toluene, methanol, ethyl alcohol, propyl alcohol, isopropanol, acetonitrile or N, N- dimethyl formyl
Amine；The reducing agent E is one of following: iron powder/concentrated hydrochloric acid, iron powder/acetic acid, palladium carbon/ammonium formate or palladium carbon/hydrazine hydrate；It is described
Iron powder/concentrated hydrochloric acid refers to that the mixing of iron powder and concentrated hydrochloric acid arbitrary proportion, iron powder/acetic acid refer to the mixed of iron powder and acetic acid arbitrary proportion
It closes, the palladium carbon/ammonium formate refers to the mixing of palladium carbon Yu ammonium formate arbitrary proportion, and the palladium carbon/hydrazine hydrate is palladium carbon and hydration
The mixture of hydrazine arbitrary proportion；
(3) compound shown in formula (V) is mixed with pivaloyl chloride or pivalic acid acid anhydride, under basic catalyst F effect, Yu Youji
In solvent G, -10~50 DEG C of fully reactings, reaction solution is post-treated, and formula (I) compound represented is made；The base catalysis
Agent F is one of following: pyridine, diethylamine, triethylamine, quinoline, N, N- dimethylaniline, 4-dimethylaminopyridine, 4- pyrrolidinyl
Pyridine or sodium carbonate；The organic solvent G is one of following: tetrahydrofuran, methylene chloride, chloroform, ethyl acetate, ether, second
Nitrile, toluene or benzene；
3. method according to claim 2, it is characterised in that: compound shown in formula (III) described in step (1) and formula (II)
The ratio between amount for the substance that feeds intake of shown compound, basic catalyst B is 1.0 ﹕, 0.8~1.2 ﹕ 1.0~8.0；The organic solvent A
Dosage 10~50mL/g is calculated as with the quality of compound shown in formula (III).
4. method according to claim 2, it is characterised in that: reaction solution described in step (1) isolates and purifies in step (1)
Method are as follows: after fully reacting, solvent is evaporated off in reaction solution, concentrate is taken to be dissolved with organic solvent C, obtains lysate, so
The column chromatography silica gel of 1.0~2.0 times of weight of concentrate is added in backward lysate, after mixing, solvent is evaporated off, it is dry, it obtains dense
Mixture is filled column, is then mixed for the petroleum ether of 1:0.1~10 with ethyl acetate with volume ratio by the mixture of contracting object and silica gel
Solution is eluant, eluent, collects the efflux containing target components, is concentrated under reduced pressure, dry, obtains formula (IV) compound represented；It is described
Organic solvent C is one of following: ethyl alcohol, chloroform, tetrahydrofuran or ethyl acetate.
5. method according to claim 2, it is characterised in that: in step (2), when the reducing agent E is iron powder/concentrated hydrochloric acid
Or iron powder/acetic acid, formula (IV) compound represented and the mass ratio that feeds intake of iron powder, concentrated hydrochloric acid or acetic acid in reducing agent E are
1.0 ﹕, 1.0~3.0 ﹕ 0.2~1.0；When the reducing agent E is palladium carbon/ammonium formate or palladium carbon/hydrazine hydrate, shown in formula (IV)
The mass ratio that feeds intake of palladium carbon, ammonium formate or hydrazine hydrate in compound and reducing agent E is 1.0 ﹕, 0.1~0.5 ﹕ 1.0~3.0.
6. method according to claim 2, it is characterised in that: step (3) carries out as follows: in -10~10 DEG C of items
Under part, into the organic solvent G solution of compound and basic catalyst F shown in formula (V) or toward compound shown in formula (V) and
The organic solvent G solution of pivaloyl chloride or pivalic acid acid anhydride is added dropwise in basic catalyst F, drop finishes, and -10~50 DEG C of reactions 3~12 are small
When, gained reaction solution is post-treated to obtain compound shown in formula (I)；Total dosage of the organic solvent G is with formula (V) shownization
The quality for closing object is calculated as 11~100mL/g.
7. method according to claim 2, it is characterised in that: compound shown in formula (V) described in step (3) and spy penta
The ratio between amount for the substance that feeds intake of acyl chlorides or pivalic acid acid anhydride, basic catalyst F is 1 ﹕, 1.0~8.0 ﹕ 1.0~3.0；It is described organic molten
The dosage of agent G is calculated as 11~100mL/g with the quality of compound shown in formula (V).
8. method according to claim 2, it is characterised in that: the method for step (3) the reaction solution post-processing are as follows: will react
Liquid filtering, filtrate steaming removal solvent take concentrate to be dissolved with organic solvent H, obtain lysate, are then added into lysate
After mixing, solvent is evaporated off in the column chromatography silica gel of 1.0~2.0 times of weight of concentrate, dry, obtains the mixing of concentrate and silica gel
Mixture is filled column by object, then using volume ratio for 1:0.1~10 petroleum ether and ethyl acetate mixture as eluant, eluent, receive
Collect the efflux containing target components, is concentrated under reduced pressure, it is dry, obtain formula (I) compound represented；The organic solvent H is following
One of: ethyl alcohol, chloroform, tetrahydrofuran or ethyl acetate.
9. pivaloyl amino benzo [d] azepine shown in formula (I) as described in claim 1Base quinazoline compounds are being made
It is standby to prevent or treat the application in human breast carcinoma drug.
10. application as claimed in claim 9, it is characterised in that the drug is with inhibition MCF-7 cell strainHJ2mm
Active drug.
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