CN104230826B - 2 fluoro quinazoline cyclics and preparation method thereof and pharmaceutical usage - Google Patents

2 fluoro quinazoline cyclics and preparation method thereof and pharmaceutical usage Download PDF

Info

Publication number
CN104230826B
CN104230826B CN201310227812.5A CN201310227812A CN104230826B CN 104230826 B CN104230826 B CN 104230826B CN 201310227812 A CN201310227812 A CN 201310227812A CN 104230826 B CN104230826 B CN 104230826B
Authority
CN
China
Prior art keywords
milligrams
alkyl
carbon atom
milliliters
benzyloxy
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CN201310227812.5A
Other languages
Chinese (zh)
Other versions
CN104230826A (en
Inventor
张倩
丁健
晋建文
谢华
董孟杰
童林江
刘振峰
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Fudan University
Shanghai Institute of Materia Medica of CAS
Original Assignee
Fudan University
Shanghai Institute of Materia Medica of CAS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Fudan University, Shanghai Institute of Materia Medica of CAS filed Critical Fudan University
Priority to CN201310227812.5A priority Critical patent/CN104230826B/en
Publication of CN104230826A publication Critical patent/CN104230826A/en
Application granted granted Critical
Publication of CN104230826B publication Critical patent/CN104230826B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/70—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings condensed with carbocyclic rings or ring systems
    • C07D239/72—Quinazolines; Hydrogenated quinazolines
    • C07D239/95—Quinazolines; Hydrogenated quinazolines with hetero atoms directly attached in positions 2 and 4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/056—Ortho-condensed systems with two or more oxygen atoms as ring hetero atoms in the oxygen-containing ring

Abstract

The invention belongs to organic chemistry and pharmaceutical synthesis field, and in particular to 2 fluoro quinazoline cyclics and preparation method thereof and pharmaceutical usage, the compound are prepared by the Jing halogen exchange reactions of 2 chloros, 2 bromos or 2 iodo quinazoline derivants；Such compound has significant receptor tyrosine kinase inhibitory activity, can be used for the medicine for preparing treatment tumour and the high expression relevant diseases of EGFR.

Description

2- fluoro quinazoline cyclics and preparation method thereof and pharmaceutical usage
Technical field
The invention belongs to organic chemistry and pharmaceutical synthesis field, and in particular to 2- fluoro quinazoline cyclics and its preparation Method and pharmaceutical usage, such compound have significant receptor tyrosine kinase inhibitory activity, can be used to prepare treatment tumour Medicine.
Background technology
Prior art discloses receptor tyrosine kinase (RTK) is that γ-phosphoric acid is transferred to albumen junket ammonia on class catalysis ATP Kinases on sour residue, can be catalyzed various substrate protein white matter tyrosine residue phosphorylations, have in cell growth, propagation, differentiation Play an important role.Research display, when this kind of kinases is over-expressed or mutation causes abnormal enzymatic activity, i.e., can cause cell Uncontrollable fast-growth, some kinases such as EGFR, VEGFR, FGFR, PDGFR, c-erbB-2, c-src, that what is waited is different Often expression is usually related to the generation of malignant tumour.Therefore those skilled in the art think appropriately to suppress the work of receptor tyrosine kinase Property be one of the effective way for treating tumour.
At present, the receptor tyrosine kinase inhibitors with quinazoline ring structure for having listed mainly have Gefitinib, Erlotinib, Lapatinib and Conmana etc., the architectural feature of these medicines are 4- aniline quinazoline ring derivatives, quinoline Oxazoline ring 2- positions atom is hydrogen atom.
The compound patent for being related to the receptor tyrosine kinase inhibitors with quinazoline ring mainly includes：
EP520722 is related to a class EGFR tyrosine kinase inhibitor 4- aniline quinazoline derivatives, and such compound has Antitumor activity：
Wherein, RaFor hydrogen atom, trifluoromethyl or nitro；RbFor hydrogen atom, trifluoromethyl or nitro；N is 1. EP635498 is related to a class EGFR tyrosine kinase inhibitor 4- aniline quinazoline derivatives, and such compound has antitumor work Property：
Wherein, R1Including the alkoxyl of hydroxyl, amino or 1-4 carbon atom, R2Including hydrogen atom, hydroxyl or halogen original Son, R3For halogen atom, n is 1,2 or 3.EP635507 is related to the tricyclic derivatives of a class EGFR tyrosine kinase inhibitor：
Wherein, R1And R2Commutable five yuan or hexatomic ring are constituted, the ring system at least contains a hetero atom；R3Including Hydrogen atom, hydroxyl or halogen atom, n are 1,2 or 3.CN1305860（Conmana patent）It is related to a class as tyrosine The quinazoline derivant for condensing of kinase inhibitor：
Wherein A is 7 to 18 yuan of rings, R1It is：Hydrogen, halogen replace or unsubstituted C1-8Alkyl, C2-8Thiazolinyl, C2-8Alkynyl, virtue Base, heteroaryl and heterocyclic radical, C1-8Alkanoyl, C1-8Alkoxycarbonyl, C1-8Alkyl sulphinyl, C1-8Alkyl sulphonyl, fragrant sulphur Acyl group, cyano group, nitro, hydroxyl, amino, carboxyl, oxo, carbamyl, C1-8Alkoxyl, C2-8Alkenyloxy group, C2-8Alkynyloxy group, C1-8 Alkylthio group, N- (C1-8Alkyl) carbamyl, bis--(C of N, N-1-8Alkyl) carbamyl, C1-8Alkanoyloxy, C1-8Alkyl amide, C3-8Alkynyl amide base, N- (C1-8Alkyl) sulfamoyl, bis--(C of N, N-1-8Alkyl) sulfamoyl；M is 0 to 3 integer；X is NR2, CHR3, O or S；The R2And R3H or C can be respectively1-8Alkyl；R be substituted or unsubstituted aryl, heterocyclic base, aryl- C1-3Alkyl and aryl-C3-7Cycloalkyl.
Present inventor intends providing a class the new 2- fluoro quinolines with notable receptor tyrosine kinase inhibitory activity Oxazoline ring derivatives, for preparing the medicine of tumour and associated receptor EGFR-TK abnormal diseases.And such chemical combination The preparation method of thing, this kind of compound have significant receptor tyrosine kinase inhibitory activity, can.
The content of the invention
It is an object of the invention to provide the new 2- fluoro quinazoline rings with notable receptor tyrosine kinase inhibitory activity Derivative compound, it is fluorine atom that its architectural feature is quinazoline ring 2- positions, and 4- positions, 6- positions and 7- positions have different replacements Base.
The 2- fluoro quinazoline cyclics of the compound the being related to structural formula as shown in Equation 1 of the present invention；
Formula 1
Wherein, X is NH, or O；
R1And R2For identical or different groups, hydrogen, amino, hydroxylamino, hydroxyl, urea groups, N- (C are respectively selected from1-8Alkyl) Amido, bis--(C of N, N-1-8Alkyl) amido, pyrrolidin-1-yl, piperidin-1-yl, piperazine -1- bases, morpholine -4- bases are thio Quinoline -4- bases, the alkyl-substituted piperazine -1- bases of 4-, halogen replacement alkylamino radical, hydroxyl alkylamino radical, alkoxyl alkylamino radical, what carboxyl replaced Alkylamino radical, the alkylamino radical that amino replaces；Amide groups containing 1-8 carbon atom, the 2- acrylamide bases containing 3-8 carbon atom, propylene Amide groups, the 2- acrylamide bases containing 3-8 carbon atom that amino replaces, bis--(C of N, N-1-8Alkyl) amido replace containing 3-8 The 2- acrylamide bases of carbon atom, benzamido, sulfoamido, benzene sulfonamido, halogenated alkyl amide base, hydroxyl is for alkane acid amides Base, alcoxyl alkyl amide, amino alkyl amide, alkylamino radical alkyl amide, di alkylamino group alkyl amide；Containing 1-8 carbon atom Oxyl, the alkoxyl containing 1-8 carbon atom, the cycloalkyl oxy of 3-8 carbon atom, halogen substituted alkoxy, what hydroxyl replaced Alkoxyl, the alkoxyl that alkoxyl replaces, the alkoxyl that amino replaces, alkylamino alkoxyl, di alkylamino group alkoxyl, piperidines- 1- base alkoxyls, morpholinyl -1- alkoxyls, piperazine -1- base alkoxyls, the alkyl-substituted piperazine -1- bases alkoxyls of 4-, pyrroles Alkane -1- base alkoxyls, trifluoromethoxy；The alkyl of 1-8 carbon atom, halohydrocarbyl, hydroxyl is for alkyl, alkoxyalkyl, hydroxyl Base alkoxyalkyl, alkyloxy-alkoxy alkyl, acyloxyalkyl, the alkyl that amino replaces, alkylamino radical alkyl, di alkylamino group hydrocarbon Base, piperidyl alkyl, morpholinyl alkyl, piperazinyl alkyl, the alkyl-substituted piperazinyl alkyl of 4-, pyrrolidinyl alkyl, the above The alkyl is the alkyl containing 1-8 carbon atom, alkylene or alkynes base；R1And R2Cyclization can be linked, composition includes 6 to 16 Yuan of rings, ring include hetero atom or without hetero atom；
R3-R5For identical or different groups, hydrogen is respectively selected from, fluorine, chlorine, bromine, nitro, hydroxyl, amino, carboxyl, 1-4 are individual Carbon atom hydrocarbyl radical, acetenyl, trifluoromethyl, trifluoromethoxy；Alkoxyl containing 1-8 carbon atom, the cycloalkanes of 3-8 carbon atom Base epoxide, aryl epoxide, heteroaromatic epoxide, heteroaromatic methoxyl group, halogenated phenoxy, a fluorophenoxy, a chlorophenoxy, Bromobenzene epoxide, phenyl polyhalide epoxide, trifluoromethyl substituent phenoxy, benzyloxy, halobenzyloxy, a fluorine benzyloxy, a benzyl chloride Epoxide, a bromo-benzyloxy, polyhalo benzyloxy, trifluoromethyl substituted benzyloxy, pyridine epoxide, haloperidid epoxide, pyridine -2- Methoxyl group, substituted pyridines -2- methoxyl groups；Formamido, carbamido group, the hydroxylamine base formamido of 1-4 carbon atom；Virtue Carbamoyl amido, fragrant miscellaneous carbamoyl amido, all kinds of substituted aryl carbamoyl amidos, all kinds of replacement heteroaromatic carbamoyls Amido, the amide groups of 1-4 carbon atom, benzamido, sulfoamido, benzene sulfonamido, halogenated alkyl amide base, hydroxyl is for alkane Amide groups, alcoxyl alkyl amide, amino alkyl amide, alkylamino radical alkyl amide, di alkylamino group alkyl amide are former containing 1-4 carbon The acyl-oxygen amino of son, containing 1-4 carbon atom alkoxyamino.
Preferably,
2- fluoro quinazoline cyclics shown in the formula 1 of the present invention,
Its architectural feature is：X is NH, or O；
R1And R2For identical or different groups, hydrogen is respectively selected from, amino, hydroxylamino, hydroxyl, urea groups are former containing 1-8 carbon The amide groups of son, the 2- acrylamide bases containing 3-8 carbon atom, acrylamido, the 2- alkene containing 3-8 carbon atom that amino replaces Amide groups, bis--(C of N, N-1-8Alkyl) amido replace the 2- acrylamide bases containing 3-8 carbon atom, sulfoamido, benzsulfamide Base, halogenated alkyl amide base, hydroxyl is for alkyl amide, alcoxyl alkyl amide, amino alkyl amide, alkylamino radical alkyl amide, dioxane Amidocyanogen amido, the oxyl containing 1-8 carbon atom, the alkoxyl containing 1-8 carbon atom, the cycloalkyl of 3-8 carbon atom Epoxide, halogen substituted alkoxy, hydroxyl substituted alkoxy, the alkoxyl that alkoxyl replaces, the alkoxyl that amino replaces, alkylamino radical alkane Epoxide, di alkylamino group alkoxyl, piperidin-1-yl alkoxyl, morpholinyl -1- alkoxyls, piperazine -1- base alkoxyls, 4- alkyl take Piperazine -1- base the alkoxyls in generation, pyrrolidin-1-yl alkoxyl, trifluoromethoxy；R1And R2Cyclization, including 6 to 16 yuan can be linked Ring, containing hetero atom or without hetero atom；
R3-R5For identical or different groups, hydrogen, fluorine, chlorine, bromine, nitro, hydroxyl, amino, 1-4 carbon atom are respectively selected from Alkyl, acetenyl, trifluoromethyl, trifluoromethoxy；Alkoxyl containing 1-8 carbon atom, the cycloalkyloxy group of 3-8 carbon atom Base, aryl epoxide, heteroaromatic epoxide, heteroaromatic methoxyl group, halogenated phenoxy, a fluorophenoxy, a chlorophenoxy, a bromobenzene Epoxide, phenyl polyhalide epoxide, trifluoromethyl substituent phenoxy, benzyloxy, halobenzyloxy, a fluorine benzyloxy, m-chloro benzyloxy, Between bromo-benzyloxy, polyhalo benzyloxy, trifluoromethyl substituted benzyloxy, pyridine epoxide, haloperidid epoxide, pyridine -2- methoxies Base, substituted pyridines -2- methoxyl groups, the amide groups of 1-4 carbon atom, formamido, carbamido group, 1-4 carbon atom Hydroxylamine base formamido；Aryl amine formamido, fragrant miscellaneous carbamoyl amido, all kinds of substituted aryl carbamoyl amidos are all kinds of Replace heteroaromatic amido formamido, sulfoamido, benzene sulfonamido.
A further object of the present invention is to provide the preparation method of above-claimed cpd, passes through 2- chloros, 2- bromines in the present invention The halogen exchange reaction of generation or 2- iodo quinazoline derivants prepares the compound；
Specifically, compound shown in formula 1 is prepared by method shown in formula 2：
Formula 2
2- chloros obtained in choosing, 2- bromos or 2- iodos quinazoline derivant are raw material；Fluorine anion（F-）It is selected from By sodium fluoride, potassium fluoride, cesium fluoride, tetrabutyl ammonium fluoride, benzyl trimethyl ammonium fluoride or the generation of tetraethyl ammonium fluoride reagent； Catalyst is selected from six oxygen -1,10- diaza-bicyclos [8,8,8] hexacosanes of 4,7,13,16,21,24-（K2.2.2, it is as follows）Deng Crown ether compound, solvent selected from acetone, acetonitrile, DMF（DMF）Or dimethyl sulfoxide (DMSO)（DMSO）Deng.
A further object of the present invention is to provide the pharmaceutical usage of above-claimed cpd, and especially the compound is used as acceptor Purposes of the tyrosine kinase inhibitor in antineoplastic is prepared,
In the present invention, the 2- fluoro quinazoline compounds of obtained formula 1, have carried out external EGFR enzymatic activitys and lung respectively The sieve test of cancer A431 cell line model drug, biological activity determination result show that described compound can significantly inhibit EGFR enzyme activity Property and tumor cell line growth, with antitumor activity, can be used to prepare the medicine of tumour, especially the high expression of EGFR Anti-tumor medicine.
Specific embodiment
In present invention experiment, the hydrogen nuclear magnetic resonance spectrum of compound (1H NMR) by Bruker-DPX (400 MHz) nuclear magnetic resonance spectrometer Determine；Mass spectrum（ESI-MS）Determined by Agilent G1946D mass spectrographs；Fusing point is determined with SGW X-4 micro-meldometers, temperature Meter is not corrected；Six oxygen -1,10- diaza-bicyclos [8,8,8] hexacosanes of 4,7,13,16,21,24-（K2.2.2, it is as follows）For moral State's ABX reagents；Other reagents are commercial reagent；Application on human skin squamous cell carcinoma A431 cells, human lung cancer PC9 cell, human lung cancer The cell lines such as A549 cells, MCF-7 Human Breast Cancer Cells and human hepatoma HepG2 cell are purchased from Unite States Standard biology product collection Center（ATCC）；Endothelial growth factor receptor（EGFR）EGFR-TK is institute of materia medica's oncology pharmacology country of the Chinese Academy of Sciences Key lab is expressed using insect baculovirus expression system, is swashed with the activated intracellular that Ni-NTA posts affinity purification is obtained Enzyme area protein tyrosine kinase, and meet requirement of experiment after testing, -70 °C of packing are preserved.
Embodiment 1 prepares the fluoro- 4- anilino-s -6,7- dimethoxyquinazolines of 2-
220 milligrams of (3.8 mMs) KF are added in 50 milliliters of three-necked bottles（Potassium fluoride, it is as follows）With 150 milligram (0.38 MM) chloro- 4- anilino- -6 of 2-, 7- dimethoxyquinazolines, nitrogen protection is lower to add 145 milligrams of catalyst K2.2.2（4, 7,13,16,21,24- six oxygen -1,10- diaza-bicyclos [8,8,8] hexacosane are as follows）(0.38 mM) and 2 milliliters DMSO（Dimethyl sulfoxide (DMSO), it is as follows）, 140 DEG C of reactions are warming up to, after 48 hours, stop reaction, after reactant liquor is cooled to room temperature, will Which is added into 40 milliliters of frozen water, separates out brown solid, and filtration drying obtains 177 milligrams of brown solid, column chromatography for separation（Chloroform: Petroleum ether:Methyl alcohol=40:10:1 wash-out）, obtain 34 milligrams of target product（Yield 29.4%）；ESI-MS m/z: 300 (M+H+); 1H NMR (CDCl3, 400 MHz) δ : 7.69 (2H, d, J = 7.93Hz, 1’-H, 5’-H), 7.45 (1H, brs, -NH), 7.42 (2H, dd, J 1 = 7.53 Hz, J 2 = 8.32 Hz, 2’-H, 4’-H), 7.20 (1H, dd, J 1= 7.14 Hz, J 2= 6.34 Hz, 3’-H), 7.15 (1H, s, 8-H), 7.04 (1H, s, 5- H), 4.00 (6H, s, 2-OCH3)。
Embodiment 2 prepares fluoro- 4- (3- the bromanilines) -6,7- dimethoxyquinazolines of 2-
Add 220 milligrams of KF (3.8 mMs) and 149 milligrams of (0.38 mM) 2- chloro- in 50 milliliters of three-necked bottles 4- (2- bromanilines) -6,7- dimethoxyquinazolines, nitrogen protection is lower to add 145 milligrams of K2.2.2 and 2 milliliter of DMSO, heats up React to 140 DEG C, stop reaction after 48 hours, after reactant liquor is cooled to room temperature, be added into into 40 milliliters of frozen water, separate out palm fibre Color solid, filtration drying obtain brown solid, column chromatography for separation（Chloroform:Petroleum ether:Methyl alcohol=40:10:1 wash-out）, obtain target product 36 milligrams of thing（Yield 25%）；
ESI-MS m/z : 378 (M+H+), 380 (M+2+H+); 1H-NMR (CDCl3, 400 MHz) δ : 7.88 (1H, t, J = 1.76 Hz, 2’-H), 7.71 (1H, dt, J 1 = 7.82 Hz, J 2 = 1.76 Hz, 6’-H), 7.32 (1H, brs, -NH-), 7.30 (1H, dd,J 1 = 7.82 Hz, J 2 = 7.44 Hz, 5’-H), 7.26 (1H, d, J = 7.44 Hz, 4’-H), 7.16 (1H, s, 8-H), 6.99 (1H, s, 5-H), 4.02 (3H, s, -OCH3), 4.01 (3H, s, -OCH3)。
Embodiment 3 prepares fluoro- 4- (3- the acetylenylanilines) -6,7- dimethoxyquinazolines of 2-
Add 220 milligrams of KF (3.8 mMs) and 128 milligrams of (0.38 mM) 2- chloro- in 50 milliliters of three-necked bottles 4- (3- acetylenylanilines) -6,7- dimethoxyquinazolines, nitrogen protection is lower to add 145 milligrams of catalyst K2.2.2(0.38 milli Mole) and 2 milliliters of DMSO, be warming up to 140 DEG C of reactions, after 48 hours, stop reaction, after reactant liquor is cooled to room temperature, be added into In 40 milliliters of frozen water, brown solid is separated out, filtration drying obtains brown solid, column chromatography for separation（Chloroform:Petroleum ether:Methyl alcohol= 40:10:1 wash-out）, obtain 37 milligrams of target product（Yield 30%）；
ESI-MS m/z : 324 (M+H+); 1H-NMR (CDCl3, 400 MHz) δ : 7.78 (1H, d, J = 8.21 Hz, 6’-H), 7.75 (1H, s, 2’-H), 7.40 (1H, brs, -NH-), 7.36 (1H, dd,J 1 = 7.82 Hz, J 2 = 7.83 Hz, 5’-H), 7.29 (1H, d, J= 7.83 Hz, 4’-H), 7.14 (1H, s, 8-H), 7.01 (1H, s, 5-H), 4.01 (3H, s, -OCH3), 3.99 (3H, s, -OCH3), 3.09 (1H, s, C≡CH)。
Embodiment 4 prepares fluoro- 4- [(3- aniline formamidos) the aniline] -6,7- dimethoxyquinazolines of 2-
Add 220 milligrams of KF (3.8 mMs) and 171 milligrams of (0.38 mM) 2- fluoro- in 50 milliliters of three-necked bottles 4- [(3- aniline formamidos) aniline] -6,7- dimethoxyquinazolines, nitrogen protection is lower to add 145 milligrams of catalyst K2.2.2 (0.38 mM) and 2 milliliters of DMSO, are warming up to 140 DEG C of reactions, stop reaction after 48 hours, after reactant liquor is cooled to room temperature, will Which is added into 40 milliliters of frozen water, separates out brown solid, and filtration drying obtains brown solid, column chromatography for separation（Chloroform:Petroleum ether: Methyl alcohol=40:10:1 wash-out）, obtain 26 milligrams of target product（Yield 16%）；
ESI-MS m/z : 434 (M+H+); 1H-NMR (CDCl3, 400 MHz) δ : 9.26 (2H, brs, - NHCONH-), 7.61-7.85 (5H, m, 2”-6”-H), 7.78 (1H, d, J = 8.21 Hz, 6’-H), 7.75 (1H, s, 2’-H), 7.40 (1H, brs, -NH-), 7.36 (1H, dd,J 1 = 7.82 Hz, J 2 = 7.83 Hz, 5’-H), 7.29 (1H, d, J= 7.83 Hz, 4’-H), 7.14 (1H, s, 8-H), 7.01 (1H, s, 5- H), 4.01 (3H, s, -OCH3), 3.99 (3H, s, -OCH3).
The fluoro- 4- of 5 2- of embodiment [(3- m-chloroaniline formamidos) aniline] -6,7- dimethoxyquinazolines
Add 220 milligrams of KF (3.8 mMs) and 184 milligrams of (0.38 mM) 2- fluoro- in 50 milliliters of three-necked bottles 4- [(3- m-chloroaniline formamidos) aniline] -6,7- dimethoxyquinazolines, nitrogen protection is lower to add 145 milligrams of catalyst K2.2.2(0.38 mM) and 2 milliliters of DMSO, are warming up to 140 DEG C of reactions, stop reaction after 48 hours, and reactant liquor is cooled to room temperature Afterwards, it is added into into 40 milliliters of frozen water, separates out brown solid, filtration drying obtains brown solid, column chromatography for separation（Chloroform:Stone Oily ether:Methyl alcohol=40:10:1 wash-out）, obtain 41 milligrams of target product（Yield 23%）；
ESI-MS m/z: 468 (M+H+); 1H-NMR (CDCl3, 400 MHz) δ : 9.22 (2H, brs, - NHCONH-), 7.81-8.05 (4H, m, 2”, 4”-6”-H), 7.78 (1H, d, J = 8.21 Hz, 6’-H), 7.75 (1H, s, 2’-H), 7.40 (1H, brs, -NH-), 7.36 (1H, dd,J 1 = 7.82 Hz, J 2 = 7.83 Hz, 5’-H), 7.29 (1H, d, J= 7.83 Hz, 4’-H), 7.14 (1H, s, 8-H), 7.01 (1H, s, 5-H), 4.01 (3H, s, -OCH3), 3.99 (3H, s, -OCH3)。
Embodiment 6 prepares fluoro- 4- (3- ethynyl phenyls) -6,7- two (2- methoxy ethoxies) quinazolines of 2-
Add 220 milligrams of KF (3.8 mMs) and 163 milligrams of (0.38 mM) 2- chloro- in 50 milliliters of three-necked bottles Bis--(2- methoxy ethoxies) quinazolines of 4- (3- acetylenylanilines) -6,7-, nitrogen protection is lower to add 145 milligrams of catalyst K2.2.2 (0.38 mM) and 2 milliliters of DMSO, is warming up to 140 DEG C of reactions, stops reaction after 48 hours, and reactant liquor is cooled to room Wen Hou, is added into into 40 milliliters of frozen water, separates out brown solid, and filtration drying obtains brown solid, column chromatography for separation（Chloroform: Petroleum ether:Methyl alcohol=40:10:1 wash-out）, obtain 31 milligrams of target product（Yield 19.8%）；
ESI-MS m/z: 434 (M+Na+); 1H-NMR (CDCl3, 400 MHz) δ : 7.82 (1H, s, NH), 7.79 (1H, d, J = 3.20 Hz, 4’-H), 7.51 (1H, s, 2’-H), 7.38 (1H, t, J = 7.60 Hz, 5’-H), 7.31 (1H, d, J = 7.60 Hz, 6’-H), 7.22 (1H, s, 5-H), 7.13 (1H, s, 8-H), 4.24-4.30 (4H, m, 2×-OCH2CH2OCH3), 3.83-3.85 (4H, m, 2×-OCH2CH2OCH3), 3.47 (6H, s, 2×-OCH3), 3.11 (1H, s, -C≡CH)。
Embodiment 7 prepares fluoro- 4- (3- bromanilines) -6,7- two (2- methoxy ethoxies) quinazolines of 2-
Add 220 milligrams of KF (3.8 mMs) and 183 milligrams of (0.38 mM) 2- chloro- in 50 milliliters of three-necked bottles Bis--(2- methoxy ethoxies) quinazolines of 4- (3- bromanilines) -6,7-, nitrogen protection is lower to add 145 milligrams of catalyst K2.2.2 (0.38 mM) and 2 milliliters of DMSO, are warming up to 140 DEG C of reactions.Stop reaction after 48 hours, after reactant liquor is cooled to room temperature, will Which is added into 40 milliliters of frozen water, separates out brown solid, and filtration drying obtains brown solid.Column chromatography for separation（Chloroform:Petroleum ether: Methyl alcohol=40:10:1 wash-out）, obtain 27 milligrams of target product（Yield 15%）.ESI-MS m/z: 466 (M+H+), 468 (M+ 2+H+); 1H-NMR (CDCl3, 400 MHz) δ : 7.90 (1H, s, 2’-H), 7.72 (1H, d, J = 7.44 Hz, 6’-H), 7.60 (1H, s, 4’-H), 7.29 (1H, t,J = 8.14 Hz, 5’-H), 7.21 (1H, s, 5-H), 7.09 (1H, s, 8-H), 4.21-4.29 (4H, m, 2×-OCH2CH2OCH3), 3.82-3.84 (4H, m, 2×-OCH2CH2OCH3), 3.35 (6H, s, 2×-OCH3).
Embodiment 8 prepares fluoro- two-(2- methoxy ethoxies) quinazolines of 4- (the fluoro- 4- bromanilines of 2-) -6,7- of 2-
Add 220 milligrams of KF (3.8 mMs) and 190 milligrams of (0.38 mM) 2- chloro- in 50 milliliters of three-necked bottles 4- (the fluoro- 4- bromanilines of 2-) -6,7- bis- (2- methoxy ethoxies) quinazoline, nitrogen protection is lower to add 145 milligrams of catalyst K2.2.2(0.38 mM) and 2 milliliters of DMSO, are warming up to 140 DEG C of reactions.Stop reaction after 48 hours, reactant liquor is cooled to room temperature Afterwards, it is added into into 40 milliliters of frozen water, separates out brown solid, filtration drying obtains brown solid.Column chromatography for separation（Chloroform:Stone Oily ether:Methyl alcohol=40:10:1 wash-out）, obtain 29 milligrams of target product（Yield 16%）.ESI-MS m/z: 484 (M+H+), 486 (M+2+H+); 1H-NMR (CDCl3, 400 MHz) δ : 8.10 (1H, s, -NH), 7.91 (1H, s, 5- H), 7.81 (1H, s, 3’-H), 7.57-7.71 (2H, m,5’-H, 6’-H), 6.95 (1H, s, 8-H), 4.28-4.30 (4H, m, 2×-OCH2CH2OCH3), 3.71-3.76 (4H, m, 2×-OCH2CH2OCH3), 3.48 (6H, s, 2×-OCH3).
The fluoro- 4- of the preparation 2- of embodiment 9 [the chloro- 4- of 3- (2- pyridomethoxies) aniline) two-(2- methoxyl group ethoxies of -6,7- Base) quinazoline
Add 220 milligrams of KF (3.8 mMs) and 207 milligrams of (0.38 mM) 2- chloro- in 50 milliliters of three-necked bottles 4- [the chloro- 4- of 3- (2- pyridomethoxies) aniline] -6,7- bis- (2- methoxy ethoxies) quinazoline, nitrogen protection is lower to add 145 Milligram catalyst K2.2.2(0.38 mM) and 2 milliliters of DMSO, are warming up to 140 DEG C of reactions.Stop reaction, reaction after 48 hours After liquid is cooled to room temperature, it is added into into 40 milliliters of frozen water, separates out brown solid, filtration drying obtains brown solid.Column chromatography point From（Chloroform:Petroleum ether:Methyl alcohol=40:10:1 wash-out）, obtain 30 milligrams of target product（Yield 15%）.ESI-MS m/z: 529 (M+H+); 1H-NMR (CDCl3, 400 MHz) δ : 8.59 (1H, d, J = 4.70 Hz, 3”-H), 7.75 (2H, t,J= 2.74 Hz, 3-H, 4’’-H), 7,65 (1H, d, J = 7.82 Hz, 6’’-H), 7.54 (1H, m, 5’’-H), 7.47 (1H, s,2’-H), 7.19 (1H, s, 5-H), 7.14 (1H, d, J = 2.73 Hz, 6’- H), 7.10 (1H, s,8-H), 7.00 (1H, d, J = 9.00 Hz, 5’-H) , 5.28 (2H, s, -CH2-), 4.21-4.27 (4H, m, 2×-OCH2CH2OCH3), 3.81-3.84 (4H, m, 2×-OCH2CH2OCH3), 3.46 (6H, s, 2×-OCH3).
Embodiment 10 prepares fluoro- 4- [the chloro- 4- of 3- (3- 4-trifluoromethylphenopendants) aniline] two (the 2- methoxyl group second of -6,7- of 2- Epoxide) quinazoline
Add 220 milligrams of KF (3.8 mMs) and 227 milligrams of (0.38 mM) 2- chloro- in 50 milliliters of three-necked bottles 4- [the chloro- 4- of 3- (3- 4-trifluoromethylphenopendants) aniline] -6,7- bis- (2- methoxy ethoxies) quinazoline, nitrogen protection is lower to be added 145 milligrams of catalyst K2.2.2(0.38 mM) and 2 milliliters of DMSO, are warming up to 140 DEG C of reactions.Stop reaction after 48 hours, After reactant liquor is cooled to room temperature, it is added into into 40 milliliters of frozen water, separates out brown solid, filtration drying obtains brown solid.Post layer Analysis is separated（Chloroform:Petroleum ether:Methyl alcohol=40:10:1 wash-out）, obtain 44 milligrams of target product（Yield 20%）.ESI-MS m/z : 582 (M+H+); 1H-NMR (DMSO-d 6, 400 MHz) δ : 10.22 (1H, s, -NH), 8.17 (1H, s, 2’’-H), 8.04 (1H, s, 2’-H), 7.86 (1H, d,J = 9.00 Hz, 4’’-H), 8.51 (1H, t, J = 8.02 Hz, 5’’-H), 7.47 (1H, d, J = 9.00 Hz, 6’’-H), 7.35 (1H, d,J = 9.00 Hz, 6’-H), 7.25 (1H, d,J = 8.61 Hz, 5’-H) , 7,21 (1H, s, 8-H), 7.16 (1H, s, 5-H), 4.27-4.29 (4H, m, 2×-OCH2CH2OCH3), 3.71-3.76 (4H, m, 2×-OCH2CH2OCH3), 3.34 (6H, s, 2×-OCH3).
Embodiment 11 prepares fluoro- 4- [the chloro- 4- of 3- (3- chlorophenoxies) the aniline] -6,7- two (2- methoxy ethoxies) of 2- Quinazoline
Add 220 milligrams of KF (3.8 mMs) and 208 milligrams of (0.38 mM) 2- chloro- in 50 milliliters of three-necked bottles 4- [the chloro- 4- of 3- (3-- chlorophenoxies) aniline] -6,7- bis- (2- methoxy ethoxies) quinazoline, nitrogen protection is lower to add 145 millis Gram catalyst K2.2.2(0.38 mM) and 2 milliliters of DMSO, are warming up to 140 DEG C of reactions.Stop reaction, reactant liquor after 48 hours After being cooled to room temperature, it is added into into 40 milliliters of frozen water, separates out brown solid, filtration drying obtains brown solid.Column chromatography for separation （Chloroform:Petroleum ether:Methyl alcohol=40:10:1 wash-out）, obtain 39 milligrams of target product（Yield 18%）.ESI-MS m/z: 548 (M +H+); 1H-NMR (CDCl3, 400 MHz) δ : 7.87 (1H, s, 2’-H), 7.68 (1H, dd,J 1 = 9.39 Hz,J 2 = 8.61 Hz, 5’’-H), 7,54 (1H, s, -NH), 7.46 (1H, s, 8-H), 7.23 (1H, d, J = 2.35 Hz, 2’’-H), 7.17 (1H, d, J = 9.39 Hz, 6’-H), 7.12 (1H, d, J = 7.82 Hz, 6’’-H), 7.08 (1H, d, J = 9.39 Hz, 5’-H), 6.95 (1H, s, 5-H), 6.87 (1H, d, J = 8.61 Hz, 4’’-H), 4.23-4.31 (4H, m, 2×-OCH2CH2OCH3), 3.83-3.85 (4H, m, 2×- OCH2CH2OCH3), 3.47 (6H, s, 2×-OCH3).
Embodiment 12 prepares fluoro- 4- (3- the acetylenylanilines) -6- acrylamido quinazolines of 2-
Add 220 milligrams of KF (3.8 mMs) and 132 milligrams of (0.38 mM) 2- chloro- in 50 milliliters of three-necked bottles 4- (3- acetylenylanilines) -6- acrylamido quinazolines, nitrogen protection is lower to add 145 milligrams of catalyst K2.2.2(0.38 milli Mole) and 2 milliliters of DMSO, it is warming up to 140 DEG C of reactions.After 48 hours stop reaction, after reactant liquor is cooled to room temperature, be added into In 40 milliliters of frozen water, brown solid is separated out, filtration drying obtains brown solid.Column chromatography for separation（Chloroform:Petroleum ether:Methyl alcohol= 40:10:1 wash-out）, obtain 37 milligrams of target product（Yield 29%）.ESI-MS m/z: 333 (M+H+);1H-NMR (DMSO-d 6, 400 MHz) δ : 10.55 (1H, s, -NH- ), 10.40 (1H, s, -NH- ), 8.88 (1H, d, J = 1.96 Hz, 5-H), 7.87 (2H, dd, J 1 = 6.26 Hz, J 2 = 3.92 Hz, 7-H, 2’-H), 7.79 (1H, d, J = 8.60 Hz, 8-H), 7.68 (1H, d, J = 9.00 Hz, 6’-H), 7.43 (1H, t, J = 8.22 Hz, 5’-H), 7.28 (1H, d, J = 7.49 Hz, 4’-H), 6.49~6.57 (1H, m, -CH), 6.32 (1H, dd, J 1 = 17.21 Hz, J 2 = 1.96 Hz, CH2=CH), 5.82 (1H, dd, J 1 = 10.17 Hz,J 2 = 1.96 Hz, CH2=CH). 3.10 (1H, s, -C≡CH).
Embodiment 13 prepares fluoro- 4- (3- the bromanilines) -6- acrylamido quinazolines of 2-
220 milligrams of KF (3.8 mMs) and 154 milligrams of (0.38 mM) chloro- 4- of 2- are added in 50 milliliters of three-necked bottles (3- bromanilines) -6- acrylamido quinazolines, nitrogen protection is lower to add 145 milligrams of catalyst K2.2.2(0.38 mM) and 2 milliliters of DMSO, are warming up to 140 DEG C of reactions.Stop reaction after 48 hours, after reactant liquor is cooled to room temperature, be added into 40 milliliters In frozen water, brown solid is separated out, filtration drying obtains brown solid.Column chromatography for separation（Chloroform:Petroleum ether:Methyl alcohol=40:10:1 Wash-out）, obtain 35 milligrams of target product（Yield 24%）.ESI-MS m/z: 389 ([M+H+2]), 387 ([M+H]);1H- NMR (DMSO-d 6, 400 MHz) δ : 10.47 (1H, s, -NH-), 10.35 (1H, s, -NH-), 8.88 (1H, d, J = 9.00 Hz, 7-H), 8.03 (1H, s, 2’-H), 7.83 (1H, dd, J 1 = 9.00 Hz,J 2 = 1.95 Hz, 7-H), 7.78 (1H, d, J = 7.04 Hz, 8-H), 7.71 (1H, d, J = 8.61 Hz, 6’-H), 7.35~7.39 (2H, m, 5’-H, 4’-H), 6.46~6.57 (1H, m, -CH), 6.32 (1H, dd,J 1 = 18.78 Hz, J 2 = 1.57 Hz, CH2=CH), 5.82 (1H, dd, J 1 = 12.13 Hz, J 2 = 1.57 Hz, CH2=CH).
Embodiment 14 prepares fluoro- 4- (the chloro- 4- fluoroanilines of the 3-) -6- acrylamido -7- methoxyquinazoline hydrochlorides of 2-
Add 220 milligrams of KF (3.8 mMs) and 155 milligrams of (0.38 mM) 2- chloro- in 50 milliliters of three-necked bottles 4- (the chloro- 4- fluoroanilines of 3-) -6- acrylamido -7- methoxyquinazoline hydrochlorides, nitrogen protection is lower to add 145 milligrams of catalyst K2.2.2 (0.38 mM) and 2 milliliters of DMSO, are warming up to 140 DEG C of reactions.Stop reaction after 48 hours, after reactant liquor is cooled to room temperature, will Which is added into 40 milliliters of frozen water, separates out brown solid, and filtration drying obtains brown solid.Column chromatography for separation（Chloroform:Petroleum ether: Methyl alcohol=40:10:1 wash-out）, obtain 33 milligrams of target product（Yield 22%）.ESI-MS m/z : 391 (M+H+); 1H-NMR (CDCl3, 400 MHz) δ : 10.12 (1H, brs, -NH), 8.20 (1H, s, 8-H), 7.78 (1H, s, 5- H), 7.28 (1H, d, J = 8.21 Hz, 6’-H), 7.75 (1H, s, 2’-H), 7.40 (1H, brs, - NH-), 7.36 (1H, dd,J=7.82 Hz, 5 '-H), 6.48-5.50 (m × 3,3H, alkene hydrogen), 3.99 (3H, s, -OCH3).
Embodiment 15 prepares fluoro- 4- (the chloro- 4- fluoroanilines of 3-) -6- [(4- dimethylaminos) but-2-enamides the base] -7- of 2- Methoxyquinazoline hydrochloride
220 milligrams of KF (3.8 mMs) and 176 milligrams of (0.38 mM) chloro- 4- of 2- are added in 50 milliliters of three-necked bottles (the chloro- 4- fluoroanilines of 3-) -6- [(4- dimethylaminos) but-2-enamides base] -7- methoxyquinazoline hydrochlorides, nitrogen protection is lower to be added 145 milligrams of catalyst K2.2.2(0.38 mM) and 2 milliliters of DMSO, are warming up to 140 DEG C of reactions.Stop reaction after 48 hours, instead After answering liquid to be cooled to room temperature, it is added into into 40 milliliters of frozen water, separates out brown solid, filtration drying obtains brown solid.Column chromatography Separate（Chloroform:Petroleum ether:Methyl alcohol=40:10:1 wash-out）, obtain 31 milligrams of target product（Yield 18%）.ESI-MS m/z: 448 (M+H+); 1H-NMR (CDCl3, 400 MHz) δ : 10.02 (1H, brs, -NH), 7.20 (1H, s, 8- H), 8.17 (1H, s, 5-H), 7.28 (1H, d, J = 8.22 Hz, 6’-H), 7.40 (1H, brs, -NH-), 7.36 (1H, dd,J=8.22 Hz, 5 '-H), the 6.75 (- H of 1H, s, 2 '), 6.78,6.40 (m, 2H, alkene Hydrogen), 3.99 (3H, s ,-OCH3), 3.02 (m, 2H, NCH2), 2.76 (s, 6H, NCH3).
Embodiment 16 prepares fluoro- 4- (the chloro- 4- bromanilines of 2-) -6- methoxyl group -7- [(1- methyl piperidine -4- bases) methoxies of 2- Base] quinazoline
220 milligrams of KF (3.8 mMs) and 193 milligrams of (0.38 mM) chloro- 4- of 2- are added in 50 milliliters of three-necked bottles (the chloro- 4- bromanilines of 2-) -6- methoxyl group -7- [(1- methyl piperidine -4- bases) methoxyl group] quinazoline, nitrogen protection is lower to add 145 Milligram catalyst K2.2.2(0.38 mM) and 2 milliliters of DMSO, are warming up to 140 DEG C of reactions.Stop reaction, reaction after 48 hours After liquid is cooled to room temperature, it is added into into 40 milliliters of frozen water, separates out brown solid, filtration drying obtains brown solid.Column chromatography point From（Chloroform:Petroleum ether:Methyl alcohol=40:10:1 wash-out）, obtain 43 milligrams of target product（Yield 23%）.ESI-MS m/z: 493 (M+H+); 1H-NMR (CDCl3, 400 MHz) δ : 7.48 (1H, s, 8-H), 7.30 (1H, brs, -NH-), 7.14 (1H, s, 5-H), 7.10 (1H, s, 3’-H), 7.06 (1H, d, J = 8.22 Hz, 5’-H), 6.86 (1H, d,J = 8.22 Hz, 6’-H), 3.96 (2H, m, -OCH2), 3.85 (3H, s, -OCH3), 2.25 (3H, s, -NCH3), 1.30-2.49 (m, 9H, H on piperidine ring).
Embodiment 17 prepares fluoro- 4- (the chloro- 4- fluoroanilines of 3-) -6- [(morpholinyl) the propoxyl group] -7- methoxyl group quinoline azoles of 2- Quinoline
220 milligrams of KF (3.8 mMs) and 182 milligrams of (0.38 mM) chloro- 4- of 2- are added in 50 milliliters of three-necked bottles (the chloro- 4- fluoroanilines of 3-) -6- [(morpholinyl) propoxyl group] -7- methoxyquinazoline hydrochlorides, 145 milligrams of the lower addition of nitrogen protection are urged Agent K2.2.2(0.38 mM) and 2 milliliters of DMSO, are warming up to 140 DEG C of reactions.Stop reaction after 48 hours, reactant liquor is cooled to After room temperature, it is added into into 40 milliliters of frozen water, separates out brown solid, filtration drying obtains brown solid.Column chromatography for separation（Chlorine It is imitative:Petroleum ether:Methyl alcohol=40:10:1 wash-out）, obtain 55 milligrams of target product（Yield 31%）.ESI-MS m/z: 465 (M+H+); 1H-NMR (CDCl3, 400 MHz) δ : 7.46 (1H, s, 8-H), 7.38 (1H, brs, -NH-), 7.26 (1H, d,J = 8.22 Hz, 6’-H), 7.24 (1H, s, 5-H), 7.06 (1H, d, J = 8.22 Hz, 5’- H), 6.80 (1H, s, 3’-H), 4.05 (2H, m, -OCH2), 3.85 (3H, s, -OCH3), 3.45-2.36 (10H, m, -NCH2, -OCH2), 1.50 (m, 2H, CH2).
Embodiment 18 prepares 12 ring of fluoro- 4- (3- acetylene aniline) four oxygen of -1,4,7,10- of 2- simultaneously [2,3-g] quinazoline
220 milligrams of KF (3.8 mMs) and 162 milligrams of (0.38 mM) chloro- 4- of 2- are added in 50 milliliters of three-necked bottles (3- acetylene aniline)-Isosorbide-5-Nitrae, 7,10- tetra- oxygen, 12 ring simultaneously [2,3-g] quinazoline, nitrogen protection is lower to add 145 milligrams of catalyst K2.2.2(0.38 mM) and 2 milliliters of DMSO, are warming up to 140 DEG C of reactions.Stop reaction after 48 hours, reactant liquor is cooled to room temperature Afterwards, it is added into into 40 milliliters of frozen water, separates out brown solid, filtration drying obtains brown solid.Column chromatography for separation（Chloroform:Stone Oily ether:Methyl alcohol=40:10:1 wash-out）, obtain 33 milligrams of target product（Yield 21%）.ESI-MS m/z: 410 (M+H+); 1H- NMR (CDCl3, 400 MHz) δ : 7.78 (1H, d,J = 8.22 Hz, 6’-H ), 7.45 (1H, s, 8-H), 7.38 (1H, brs, -NH-), 7.26 (), 7.26 (1H, s, 5-H), 7.06 (1H, dd, J = 8.22, 7.86 Hz, 5’-H), 6.80 (1H, d, J = 7.86 Hz, 5’-H), 4.15 (4H, m, -OCH2×2), 3.84 (4H, m, -OCH2×2), 3.45 (4H, m, -OCH2×2), 2.50 (1H, s, CH), 3.10 (1H, s, -C≡ CH).
19 biological activity determination of embodiment
1. cell proliferation experiment detection method is adopted：
Sulphonyl rhodamine B（Sulforhodamine B, SRB）Method：Difference of the certain amount in exponential phase is swollen Oncocyte is inoculated in 96 well culture plates, after 24 h cell attachments of culture, adds the test-compound of the present invention of variable concentrations, often Individual concentration sets three wells, and sets the DMSO solution control of respective concentration and acellular zeroing hole, 72 h of treated with medicaments cell Afterwards, incline nutrient solution, adds the 10% of 100 μ L ice precoolings solution of trichloroacetic acid to fix cell, uses distilled water after 4 °C of placement 1h Washing 5 times, spontaneously dries in air, is subsequently adding 100 mL SRB（4 mg/mL）（Sigma, St Louis, MO, USA）It is molten Liquid, dyes 15 min, removes dyeing liquor in room temperature, washed 5 times with 1 % glacial acetic acid, be air-dried, and is eventually adding 150 μ L, 10 mM Tris solution (pH 10.5), wavelengthtunable declines orifice plate ELIASA (VERSAmax, Molecular Device Corporation, Sunnyvale, CA, USA) OD values are determined under 515 nm wavelength, medicine is calculated to thin with following equation The inhibiting rate of intracellular growth：Inhibiting rate（%）=（OD compares-OD dosings）/ OD controls × 100%.
2. kinase assay experimental technique is adopted：Enzyme linked immunosorbent assay（ELISA）：Enzyme reaction substrate Poly (Glu, Tyr)4:120 μ g/mL are diluted to the PBS without potassium ion, coated elisa plate puts 37 °C overnight, during reaction, use is initially charged per hole The 80 μ L of ATP solution (final concentration of 5 μM of ATP) of reaction buffer dilution, are subsequently adding the test-compound of gradient concentration Or DMSO solution（Negative control）10 μ L, are finally separately added into the 10 μ L of EGFR-TK of reaction buffer dilution to start Reaction, puts 37 °C of shaking tables and reacts 1 h, after reaction terminates, T-PBS board-washings three times.Then 100 μ L PY99 antibody are added per hole （Antibody is diluted with the T-PBS containing BSA 5mg/ml）, 37 °C of shaking tables 0.5 h of reaction.T-PBS board-washings three times, add horseradish peroxide Compound enzyme marks the 100 mL/ holes of IgG of sheep anti mouse（Antibody is diluted with the T-PBS containing BSA 5mg/mL）, 37 °C of shaking table reactions 0.5 h.T-PBS board-washings three times, are subsequently adding the 100 mL/ holes of OPD nitrite ions of 2mg/mL, and 25 °C of lucifuges react 1-10 min. Add 2 M H2SO450 mL/ holes stopped reactions, are declined orifice plate ELIASA VERSAmax readings with wavelengthtunable, and wavelength is 492 nm.The inhibiting rate of sample is tried to achieve by following equation：
Experiment is repeated 3 times the above, averages.
Active determination in vitro result shows, the compound of the present invention can significantly inhibit EGFR enzymatic activitys and tumor cell line Growth, with antitumor activity, described compound can be used for the medicine for preparing tumour, and especially the high expression of EGFR is swollen Knurl medicine.
Table 1 is the initial in vitro Activity Results of embodiment compound.
Table 1
Embodiment 10μM is to EGFR tyrosine kinase activities inhibiting rate mean value (%) 10μM is to A431 cell lines proliferation inhibition rate mean value (%)
Erlotinib 85.9 45.1
Embodiment 1 2.3 17.0
Embodiment 2 79.8 51.8
Embodiment 3 74.0 61.7
Embodiment 4 77.6 68.9
Embodiment 5 86.1 72.3
Embodiment 6 45.0 49.1
Embodiment 7 62.7 68.6
Embodiment 8 67.8 53.2
Embodiment 9 84.7 51.4
Embodiment 10 75.9 72.3
Embodiment 11 72.6 55.6
Embodiment 12 89.4 71.3
Embodiment 13 89.3 74.6
Embodiment 14 80.2 77.3
Embodiment 15 86.9 68.5
Embodiment 16 84.2 59.4
Embodiment 17 75.2 58.5
Embodiment 18 83.1 61.2
20 reference examples of embodiment：
1.2- bromines quinazoline derivant and 2- iodine quinazoline derivants：In the class formation, carbon-bromine key and carbon-iodine bond have Stronger reactivity, compound are very unstable, without druggability；
2.2- chloroquinazoline derivatives are with reference to the synthesis of 3 route of formula：
Formula 3
The biologically active check experiment Activity Results of 3.2- Fluquinconazoles quinoline derivant and 2- chloroquinazoline derivatives show, 2- The anti-tumour cell proliferative activity of Fluquinconazole quinoline derivant is significantly higher than corresponding 2- chloroquinazoline derivatives；
Control compound is selected from：
Table 2 is the anti-tumour cell proliferative activity of part 2- chlorine and 2- Fluquinconazole quinoline derivants.
Table 2

Claims (1)

1. a kind of preparation method of 2- fluoro quinazoline compounds, it is characterised in that by route shown in formula 2,
X is NH or O；
R1And R2For identical or different groups, hydrogen, N- (C are respectively selected from1-8Alkyl) amido, bis--(C of N, N-1-8Alkyl) amido, 2- acrylamide bases containing 3-8 carbon atom, the 2- acrylamide bases containing 3-8 carbon atom that amino replaces, the alkane of 1-8 carbon atom Epoxide, piperidin-1-yl alkoxyl, morpholinyl -1- alkoxyls, the alkyl of 1-8 carbon atom, halohydrocarbyl, alkyloxy-alkoxy Alkyl；
R3-R5For identical or different groups, hydrogen, fluorine, chlorine, bromine, acetenyl are respectively selected from；Alkoxyl containing 1-8 carbon atom, Between fluorophenoxy, a chlorophenoxy, a bromobenzene epoxide, m-trifluoromethyl phenoxy group, benzyloxy, a fluorine benzyloxy, m-chloro benzyloxy Base, a bromo-benzyloxy, m-trifluoromethyl benzyloxy, pyridine -2- methoxyl groups；
Wherein, with 2- chloro quinazoline derivatives derivatives as initiation material；With sodium fluoride, potassium fluoride, cesium fluoride, tetrabutyl ammonium fluoride, Benzyl trimethyl ammonium fluoride or tetraethyl ammonium fluoride are that fluorination reagent generates fluorine anion；Catalyst is selected from 4,7,13,16, 21,24- six oxygen -1,10- diaza-bicyclos [8,8,8] hexacosane crown ether compound, solvent selected from acetone, acetonitrile, N, N- bis- NMF or dimethyl sulfoxide (DMSO).
CN201310227812.5A 2013-06-08 2013-06-08 2 fluoro quinazoline cyclics and preparation method thereof and pharmaceutical usage Expired - Fee Related CN104230826B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201310227812.5A CN104230826B (en) 2013-06-08 2013-06-08 2 fluoro quinazoline cyclics and preparation method thereof and pharmaceutical usage

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201310227812.5A CN104230826B (en) 2013-06-08 2013-06-08 2 fluoro quinazoline cyclics and preparation method thereof and pharmaceutical usage

Publications (2)

Publication Number Publication Date
CN104230826A CN104230826A (en) 2014-12-24
CN104230826B true CN104230826B (en) 2017-03-29

Family

ID=52219834

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201310227812.5A Expired - Fee Related CN104230826B (en) 2013-06-08 2013-06-08 2 fluoro quinazoline cyclics and preparation method thereof and pharmaceutical usage

Country Status (1)

Country Link
CN (1) CN104230826B (en)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105777656B (en) * 2014-12-25 2018-02-02 深圳海王医药科技研究院有限公司 Nabumetone replaces the beta crystal and preparation method and the pharmaceutical composition containing it of Buddhist nun's tosilate
CN105777655B (en) * 2014-12-25 2018-02-02 深圳海王医药科技研究院有限公司 Nabumetone replaces the alpha-crystal form and preparation method and the pharmaceutical composition containing it of Buddhist nun's tosilate
CN105384745A (en) * 2015-07-27 2016-03-09 北京师范大学 Crown ether cyclic quinazoline compound, preparation method therefor and application thereof in preparing tumor therapy and imaging drug
CN107501283B (en) * 2017-09-13 2020-06-02 北京师范大学 Preparation of substituted arylmethyl hetero-substituted anilino ethylene glycol ether cycloquinazoline and application of tumor treatment drug
CN110372666B (en) * 2018-04-13 2022-11-08 华东理工大学 Quinazoline compound as EGFR (epidermal growth factor receptor) triple mutation inhibitor and application thereof
CN110862398B (en) * 2018-08-27 2021-04-06 北京赛特明强医药科技有限公司 Urea substituted aromatic ring-linked dioxane quinazoline or quinoline compound, composition and application thereof

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NZ243082A (en) * 1991-06-28 1995-02-24 Ici Plc 4-anilino-quinazoline derivatives; pharmaceutical compositions, preparatory processes, and use thereof
US7078409B2 (en) * 2002-03-28 2006-07-18 Beta Pharma, Inc. Fused quinazoline derivatives useful as tyrosine kinase inhibitors
US8575339B2 (en) * 2011-07-05 2013-11-05 Xueheng Cheng Derivatives of erlotinib

Also Published As

Publication number Publication date
CN104230826A (en) 2014-12-24

Similar Documents

Publication Publication Date Title
CN104230826B (en) 2 fluoro quinazoline cyclics and preparation method thereof and pharmaceutical usage
US6608048B2 (en) Tricyclic protein kinase inhibitors
Bold et al. New anilinophthalazines as potent and orally well absorbed inhibitors of the VEGF receptor tyrosine kinases useful as antagonists of tumor-driven angiogenesis
TWI233437B (en) Substituted 3-cyanoquinolines
KR100832593B1 (en) Quinazoline derivatives as an signal trnasduction inhibitor and method for the preparation thereof
Chang et al. Development of a series of novel 4-anlinoquinazoline derivatives possessing quinazoline skeleton: Design, synthesis, EGFR kinase inhibitory efficacy, and evaluation of anticancer activities in vitro
EP3299369A1 (en) Pyrido-azaheterecydic compound and preparation method and use thereof
Mghwary et al. Novel thienopyrimidine derivatives as dual EGFR and VEGFR-2 inhibitors: design, synthesis, anticancer activity and effect on cell cycle profile
Wang et al. Design, synthesis, biological evaluation and molecular docking study of novel thieno [3, 2-d] pyrimidine derivatives as potent FAK inhibitors
EP1268487B1 (en) Tricyclic protein kinase inhibitors
CN106029646A (en) Pyrimidine-2,4-diamine derivative and pharmaceutical anticancer composition containing same as active ingredient
Zhou et al. Design, synthesis and pharmacological evaluation of 6, 7-disubstituted-4-phenoxyquinoline derivatives as potential antitumor agents
CN102875570A (en) Quinazoline derivative, and preparation method and application thereof
Yin et al. Design, synthesis and biological activities of novel oxazolo [4, 5-g] quinazolin-2 (1H)-one derivatives as EGFR inhibitors
Tang et al. Synthesis and antiproliferative activity of 6, 7-disubstituted-4-phenoxyquinoline derivatives bearing the 2-oxo-4-chloro-1, 2-dihydroquinoline-3-carboxamide moiety
Zhang et al. Discovery of a potent dual EGFR/HER-2 inhibitor L-2 (selatinib) for the treatment of cancer
Nan et al. Design, synthesis and evaluation of sulfonylurea-containing 4-phenoxyquinolines as highly selective c-Met kinase inhibitors
Elbadawi et al. 2-Arylquinolines as novel anticancer agents with dual EGFR/FAK kinase inhibitory activity: synthesis, biological evaluation, and molecular modelling insights
Upadhyay et al. Evaluation of Pyrano [3, 2 C] quinoline analogues as anticancer agents
Lu et al. Design and synthesis of novel benzo [d] oxazol-2 (3H)-one derivatives bearing 7-substituted-4-enthoxyquinoline moieties as c-Met kinase inhibitors
Ismail et al. Synthesis, antimicrobial activity and molecular modeling study of substituted 5-aryl-pyrimido [5, 4-c] quinoline-2, 4-diones
CN111356687B (en) Pyrimidine-containing tri-substituted imidazole compound and application thereof
AU2020288706A1 (en) Benzo[h][1,6] naphthyridin-2(1H)-ones as BMX inhibitors, for use against cancer
CN104230922A (en) Five-membered heterocycle-pyridine compound as well as preparation method and application thereof
Ghannam et al. Imidazo [4, 5-b] phenazines as dual topoisomerase I/IIα inhibitors: design, synthesis, biological evaluation and molecular docking

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20170329

Termination date: 20200608

CF01 Termination of patent right due to non-payment of annual fee