CN103664991A - Thiophene [2, 3-d] pyrimidine derivative as well as preparation method and use thereof - Google Patents
Thiophene [2, 3-d] pyrimidine derivative as well as preparation method and use thereof Download PDFInfo
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- CN103664991A CN103664991A CN201210349508.3A CN201210349508A CN103664991A CN 103664991 A CN103664991 A CN 103664991A CN 201210349508 A CN201210349508 A CN 201210349508A CN 103664991 A CN103664991 A CN 103664991A
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- 0 Cc1c[s]c2ncnc(OCC(O[C@@](*)C3)=CC3c3ccc(C)cc3)c12 Chemical compound Cc1c[s]c2ncnc(OCC(O[C@@](*)C3)=CC3c3ccc(C)cc3)c12 0.000 description 3
- CSLNJYQGBWVCBR-UHFFFAOYSA-N CC(C)(C)c1cc(C(Cl)=NCN2)c2[s]1 Chemical compound CC(C)(C)c1cc(C(Cl)=NCN2)c2[s]1 CSLNJYQGBWVCBR-UHFFFAOYSA-N 0.000 description 1
- VNTXXAMIQFTECR-UHFFFAOYSA-N CCc([o]c(COc1c(c(C)c[s]2)c2ncn1)c1)c1-c(cc1)ccc1OC Chemical compound CCc([o]c(COc1c(c(C)c[s]2)c2ncn1)c1)c1-c(cc1)ccc1OC VNTXXAMIQFTECR-UHFFFAOYSA-N 0.000 description 1
- SNHXPOZHZQCRTD-UHFFFAOYSA-N CCc([o]c(COc1ncnc2c1c(C)c[s]2)c1)c1-c(cc1)ccc1F Chemical compound CCc([o]c(COc1ncnc2c1c(C)c[s]2)c1)c1-c(cc1)ccc1F SNHXPOZHZQCRTD-UHFFFAOYSA-N 0.000 description 1
- XRHPZMRUAYJHJC-UHFFFAOYSA-N Cc1c[s]c2c1c(OCc1cc(-c3ccccc3Cl)n[o]1)ncn2 Chemical compound Cc1c[s]c2c1c(OCc1cc(-c3ccccc3Cl)n[o]1)ncn2 XRHPZMRUAYJHJC-UHFFFAOYSA-N 0.000 description 1
- VTWKXBJHBHYJBI-VURMDHGXSA-N O/N=C\c1ccccc1 Chemical compound O/N=C\c1ccccc1 VTWKXBJHBHYJBI-VURMDHGXSA-N 0.000 description 1
- CITYOBPAADIHAD-UHFFFAOYSA-N OCc1cc(-c2ccccc2)n[o]1 Chemical compound OCc1cc(-c2ccccc2)n[o]1 CITYOBPAADIHAD-UHFFFAOYSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
- C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
- C07D495/04—Ortho-condensed systems
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Abstract
The invention provides a thiophene [2, 3-d] pyrimidine derivative containing isoxazole heterocycle shown by formula (I) or a pharmaceutically accepted salt thereof, wherein R1 and R2 are respectively selected from hydrogen, C1-6 alkyl, C1-6 alkoxy, halogenated C1-6 alkyl, halogenated C1-6 alkoxy, aryl optionally substituted by R<7> or heteroaryl optionally substituted by R<8>; or R1, R2 and carbon atoms connected with R1 and R2 form 4-membered to 6-membered carbocycle or heterocycle; Z is -NR5-, C(R6)2, -S- or -O-; R3 is selected from hydrogen, halogen, C1-6 alkyl, C1-6 alkoxy, halogenated C1-6 alkyl or halogenated C1-6 alkoxy; n is an integer from 0 to 5; R4 is selected from hydrogen, C1-6 alkyl, c1-6 alkoxy, halogenated C1-6 alkyl, aryl optionally substituted by R<9> or heteroaryl optionally substituted by R<10>. The invention further provides a preparation method and medical use of the compound represented by the formula (I) as well as the pharmaceutically accepted salt thereof; the compound can act as a medicine or a lead compound for treating tumor, cancer and other diseases.
Description
Technical field
The present invention relates to the thiophene [2 of the Han isoxazole heterocycle of a class formation novelty, 3-d] pyrimidine derivatives and containing the medical composition and its use of this analog derivative, particularly: such thiophene [2,3-d] pyrimidine derivatives or its pharmaceutical composition have inhibition colorectal cancer cell lines (HCT-116) and human lung carcinoma cell line (A549), can be used as medicine or the lead compound of the diseases such as the treatment tumour relevant with Tyrosylprotein kinase, cancer.
Background technology
Urogastron (Epidermal Growth Factor, EGF) with EGF-R ELISA (Epidermal Growth Factor Receptor, EGFR) in conjunction with the activity that can activate Tyrosylprotein kinase, and therefore activate the reaction that causes cell proliferation.The overexpression of EGFR and increased activity all will cause uncontrollable cell fission.
Epidermal growth factor recipient tyrosine kinase (EGFR-TK) is the protein tyrosine kinase of finding the earliest, be distributed in widely on the cytolemma of each tissue of human body, it crosses and expresses in most of tumours (as: bladder cancer, nonsmall-cell lung cancer, ovarian cancer, mammary cancer, cancer of the stomach, esophagus cancer etc.), the intracellular region of EGFR has the binding site of Triphosaden (ATP), EGFR inhibitor can emulatively combine with ATP-binding site, thereby the phosphorylation that suppresses EGFR, the conduction of blocking-up downstream signal, and then the growth of inhibition tumor cell, differentiation and transfer.The EGFR acceptor of take is one of the very active field of studying in current cancer therapy as target spot carries out targeting therapy on tumor, has also obtained significant curative effect in clinical study.
Patent WO2009/104027 discloses a series of thiophene [2,3-d] pyrimidine derivates and has had inhibition tyrosine kinase activity; The 4-bit strip that patent WO2009/104026 discloses thiophene [2,3-d] pyrimidine has some thiophene [2,3-d] pyrimidine derivatives of phenylamino replacement to have antitumour activity.Above-mentioned document is incorporated herein by reference.
The present invention is based on patent WO2009/104027 and patent WO2009/104026 introduces thiophene [2 Jiang isoxazole heterocycle, 3-d] pyrimidine parent nucleus, the thiophene [2 that has synthesized a series of Han isoxazole heterocycles, 3-d] pyrimidine derivatives, vitro inhibition colorectal cancer cell lines (HCT-116) and human lung carcinoma cell line (A549) activity show that this compounds is 1 * 10
-4under M concentration, there is stronger colorectal cancer cell lines (HCT-116) and human lung carcinoma cell line (A549) activity of pressing down.Can be used as candidate compound or lead compound anticancer, tumour medicine.
Summary of the invention
The object of the invention is to, thiophene [2, the 3-d] pyrimidine compound suc as formula the Han isoxazole heterocycle shown in (I) is provided.Show through activity research, using this compound and there is as activeconstituents colorectal cancer cell lines (HCT-116) and human lung carcinoma cell line (A549) activity of pressing down.The present invention is achieved through the following technical solutions:
Thiophene [2, the 3-d] pyrimidine derivatives of the Han isoxazole heterocycle shown in a kind of formula (I), or its pharmacy acceptable salt, or solvate:
Wherein: R
1and R
2can be identical or different, be selected from independently of one another hydrogen, C
1-6alkyl, C
1-6alkoxyl group, halo C
1-6alkyl, halo C
1-6the C that alkoxyl group or hydroxyl replace
1-6alkyl, optionally by R
7the aryl replacing or optionally by R
8the heteroaryl replacing;
Or R
1and R
2coupled carbon atom forms 4 to 6 yuan of carbocyclic rings or heterocycles together, and described carbocyclic ring or heterocycle are optionally by R
11replace; Described heterocycle contains at least 1 and is selected from N, the heteroatoms of O or S;
Z is-NR
5-, C (R
6)
2,-S-or-O-, wherein R
5for hydrogen or C
1-6alkyl, R
6identical or different, be selected from hydrogen, C
1-6alkyl, halo C
1-6the C that alkyl or hydroxyl replace
1-6alkyl;
R
3be selected from hydrogen, halogen, C
1-6alkyl, C
1-6alkoxyl group, halo C
1-6alkyl, halo C
1-6the C that alkoxyl group or hydroxyl replace
1-6alkyl, the integer that n is 0 ~ 5;
R
4be selected from hydrogen, C
1-6alkyl, C
1-6alkoxyl group, halo C
1-6alkyl or halo C
1-6alkoxyl group, optionally by R
9the aryl replacing or optionally by R
10the heteroaryl replacing;
R
7, R
8, R
9, R
10or R
11be selected from independently of one another hydrogen, hydroxyl, sulfydryl, cyano group, amino, C
1-6alkylamino, two (C
1-6alkyl) amino, nitro, halogen, C
1-6alkyl, C
1-6alkoxyl group, C
1-6alkylthio, carboxyl, halo C
1-6alkyl, halo C
1-6alkoxyl group, halo C
1-6alkylthio, the C that hydroxyl replaces
1-6alkyl.
According to the preferred technical solution of the present invention, in formula (I):
R
1and R
2can be identical or different, be selected from independently of one another hydrogen, C
1-6alkyl, C
1-6alkoxyl group, halo C
1-6alkyl, halo C
1-6alkoxyl group, aryl or by R
7the aryl replacing, or
R
1and R
2coupled carbon atom forms 4 to 6 yuan of carbocyclic rings together, and described carbocyclic ring is optionally by hydrogen, C
1-6alkyl, halogen, nitro or amino replacement;
Z is-NR
5-, C (R
6)
2,-S-or-O-, wherein R
5for hydrogen or C
1-3alkyl, R
6identical or different, be selected from hydrogen, C
1-3alkyl, or the C that replaces of hydroxyl
1-3alkyl;
R
3be selected from hydrogen, halogen, C
1-6alkyl, C
1-6alkoxyl group or halo C
1-6alkyl or halo C
1-6alkoxyl group, the integer that n is 0-5;
R
4be selected from hydrogen, C
1-6alkyl, C
1-6alkoxyl group, halo C
1-6alkyl, aryl or by R
9the aryl replacing.
According to the preferred technical solution of the present invention, in formula (I):
R
1and R
2can be identical or different, be selected from independently of one another hydrogen, C
1-6alkyl, C
1-6alkoxyl group, halo C
1-6alkyl, halo C
1-6alkoxyl group, phenyl or by R
7the phenyl replacing, or
Or R
1and R
2coupled carbon atom forms 4 to 6 yuan of carbocyclic rings together;
R
4be selected from hydrogen, C
1-6alkyl, C
1-6alkoxyl group, halo C
1-6alkyl, phenyl or by R
9the phenyl replacing.
According to preferred version of the present invention, R
1and R
2be selected from independently of one another hydrogen, C
1-3alkyl, phenyl or by R
7the phenyl replacing, more preferably hydrogen, methyl, ethyl, propyl group, sec.-propyl, butyl, isobutyl-, the tertiary butyl or phenyl;
Z is-NH-, CH
2or-O-;
R
3be selected from hydrogen, fluorine, chlorine, bromine, methyl, methoxyl group or trifluoromethyl, n is preferably 1-4, more preferably 2-3.
R
4be selected from hydrogen, C
1-6alkyl, C
1-6alkoxyl group or halo C
1-6alkyl, phenyl or substituted-phenyl.
According to more preferably scheme of the present invention, R
1and R
2can be identical or different, be selected from independently of one another C
1-6alkyl, phenyl or by R
7the phenyl replacing, more preferably hydrogen, methyl, ethyl, the tertiary butyl, phenyl;
Z is-NH-or-O-,
R
3preferably ortho position or the contraposition of isoxazole ring, more preferably 4-fluorine, 4-chlorine, 2-chlorine, 4-bromine, 2,4-dichloro, 4-methyl, 4-methoxyl group, hydrogen, 4-trifluoromethyl or 2,4-dimethoxy;
R
4be selected from hydrogen, methyl or phenyl.
According to the present invention, more preferably R
1be selected from hydrogen, methyl, phenyl; R
2be selected from hydrogen, methyl, ethyl or the tertiary butyl.
According to the preferred technical solution of the present invention, wherein:
R
7, R
8, R
9or R
10be selected from independently of one another hydrogen, hydroxyl, cyano group, amino, C
1-6alkylamino, two (C
1-6alkyl) amino, halogen, C
1-6alkyl, C
1-6alkoxyl group, C
1-6alkylthio, carboxyl, halo C
1-6alkyl, halo C
1-6alkoxyl group or for C
1-6alkylthio.
Further, R
7, R
8, R
9or R
10be selected from independently of one another hydrogen, hydroxyl, sulfydryl, cyano group, amino, halogen, C
1-6alkyl, C
1-6alkoxyl group, C
1-6alkylthio, carboxyl.Further, R
7, R
8, R
9or R
10be selected from independently of one another hydrogen, hydroxyl, cyano group, amino, halogen, C
1-6alkyl, C
1-6alkoxyl group, C
1-6alkylthio or carboxyl.
According to the present invention, thiophene [2, the 3-d] pyrimidine compound shown in described formula (I) is selected from following any compound:
Thiophene [2, the 3-d] pyrimidine compound of the Han isoxazole heterocycle shown in formula (I), can select respectively to form pharmacy acceptable salt with pharmaceutically acceptable acid.Wherein term " pharmacy acceptable salt " includes but not limited to the salt forming with mineral acid, example hydrochloric acid salt, phosphoric acid salt, diphosphate, hydrobromate, vitriol,-sulfinate, nitrate and class thereof are saloid, preferably salt hydrochlorate, nitrate, vitriol or phosphoric acid salt.Also comprise the salt forming with organic acid, as lactic acid salt, oxalate, malate, maleate, fumarate, tartrate, succinate, Citrate trianion, sulfonate, tosilate, 2-isethionate, benzoate, salicylate, stearate, trifluoroacetic acid or amino acid; Alkanoate is as formate, acetate etc., or HOOC-(CH
2) n-COOH wherein n be the salt of 0-4, and class is saloid.Preferred organic acid salt is selected from formate, acetate, oxalate, Citrate trianion, fumarate, maleate, malate, lactic acid salt, tartrate, tosilate, trifluoroacetate or amino acid salts.Similarly, pharmaceutically acceptable positively charged ion includes but not limited to sodium, potassium, calcium, aluminium, lithium and ammonium.
Term " solvate " comprises hydrate and alcohol adduct.
The present invention also provides a kind of pharmaceutical composition, it comprises the thiophene [2 shown in aforesaid formula (I), 3-d] pyrimidine compound, or its pharmacy acceptable salt or solvate, and at least one pharmaceutically acceptable, inertia, nontoxic vehicle or carrier or thinner.
Pharmaceutical composition according to described, is characterized in that, described pharmaceutical composition also comprises the acceptable subsidiary material of one or more pharmacy that are selected from weighting agent, disintegrating agent, lubricant, glidant, effervescent, correctives, sanitas and coating material.
The present invention also provides a kind of pharmaceutical preparation, it comprises the thiophene [2 shown in aforesaid formula (I), 3-d] pyrimidine compound, or its pharmacy acceptable salt or solvate, and at least one pharmaceutically acceptable, inertia, nontoxic vehicle or carrier or thinner.
According to pharmaceutical preparation of the present invention, it is characterized in that, described preparation is preferably solid orally ingestible, liquid oral medicine or injection.
According to pharmaceutical preparation of the present invention, described preparation is selected from tablet, dispersible tablet, enteric coated tablet, chewable tablet, orally disintegrating tablet, capsule, granule, oral solution, injection liquid drugs injection, injection freeze-dried powder, infusion solutions or primary infusion.
The present invention also provides thiophene [2,3-d] pyrimidine compound or its pharmacy acceptable salt shown in the formula (I) of a kind of claim 1-4 as medicine, especially a kind of medicine that is effective to treat lesion/cancer disease.
The present invention also provides as the thiophene [2 shown in the formula of the aforementioned any one of medicine (I), 3-d] pyrimidine compound or its pharmacy acceptable salt or solvate, especially a kind of being used for the treatment of for the medicine that suppresses EGFR transition expression and/or the effective tumour of hyperactivity.
The present invention also provides thiophene [2, the 3-d] pyrimidine compound shown in a kind of formula (I) of aforementioned any one or its pharmacy acceptable salt or solvate in the application for the preparation of in antitumor or cancer drug.
According to application of the present invention, wherein said tumour or cancer are selected from: bladder cancer, nonsmall-cell lung cancer, ovarian cancer, mammary cancer, cancer of the stomach, esophagus cancer, lung cancer, head and neck cancer, colorectal carcinoma, pharynx cancer and carcinoma of the pancreas etc., more preferably nonsmall-cell lung cancer.
According to the present invention, also provide thiophene [2, the 3-d] pyrimidine compound shown in the formula (I) of any one in a kind of claim 1-8 and/or pharmacy acceptable salt to suppress the application in the transition expression of EGFR and/or the inhibitor of hyperactivity in preparation.
The present invention also provides thiophene [2, the 3-d] pyrimidine compound shown in a kind of formula (I) of aforementioned any one and/or pharmacy acceptable salt or solvate in the application for the preparation of pressing down in the medicine of colorectal cancer cell lines (HCT-116) and human lung carcinoma cell line (A549) activity.
The present invention also provides the preparation method of thiophene [2, the 3-d] pyrimidine compound of the Han isoxazole heterocycle shown in a kind of formula (I), it is characterized in that, described method comprises the steps:
Shown in formula II 5, the chloro-thiophene [2 of 6-bis-replacement-4-, 3-d] 3-substituted-phenyl-5-methylol-isoxazoles shown in pyrimidine and formula III or 3-substituted-phenyl-5-aminomethyl-isoxazoles shown in formula IV, in dry organic solvent and alkaline acid binding agent system, reaction prepares the compound shown in formula (I-1) or formula (I-2):
Meanwhile, for Z, be other substituent formula (I) compound, for example R
5for other substituent-NR
5, C (R
6)
2, during S, can adopt (II) and-NH(R
5) or Cl-C (R
6)
2or 3-substituted-phenyl-5-sulfydryl isoxazole prepared by corresponding linked reaction, wherein 3-substituted-phenyl-5-sulfydryl isoxazole be take propine mercaptan as raw material, the building-up process preparation of employing formula (III).
According to the present invention, described temperature of reaction be-20 ℃ to reflux conditions, preferably room temperature is to reflux conditions.
According to the present invention, described organic solvent is aromatic hydrocarbons, halohydrocarbon, C
1-C
6lower alcohols, tetrahydrofuran (THF) or dimethyl sulfoxide (DMSO) (DMF).Preferably, described solvent is selected from benzene,toluene,xylene, methylene dichloride, chloroform, Virahol, tetrahydrofuran (THF) or DMF, more preferably Virahol.
According to the present invention, described alkaline acid binding agent is organic bases or mineral alkali, and described organic bases is selected from triethylamine, tripropyl amine, DMAP, potassium tert.-butoxide etc.; Described mineral alkali is selected from salt of wormwood, sodium hydride, sodium carbonate etc.Preferred acid binding agent is triethylamine.
According to the present invention, intermediate 3-substituted-phenyl 5-methylol-isoxazoles of described formula (III) are prepared by following method:
(1) substituted benzaldehyde and azanol or hydroxylamine hydrochloride are in methanol/water system, and under the catalysis of sodium carbonate, reaction generates corresponding benzaldoxime;
(2) benzaldoxime step (1) being made and propiolic alcohol form formula (III) isoxazole compound by 1,3-Dipolar Cycloaddition under the effect of N-bromo-succinimide (NCS) and alkaline acid binding agent.
According to the present invention, preferably, described alkaline acid binding agent is selected from organic bases or mineral alkali, and described organic bases is selected from triethylamine, tripropyl amine, DMAP, DMF, N-methylmorpholine etc.; Described mineral alkali is selected from salt of wormwood, sodium hydride, sodium carbonate etc.Preferred alkaline acid binding agent is triethylamine.
Described temperature of reaction is 0 to spend to reflux temperature, is preferably room temperature to reflux temperature.
According to the present invention, intermediate 3-substituted-phenyl 5-aminomethyl-isoxazoles of described formula (IV) are prepared by following method:
(3) formula (III) isoxazole compound (preferably obtaining by preceding method) reacts with SULPHURYL CHLORIDE and obtains formula V compound;
According to the preferred technical solution of the present invention, in step (3), described SULPHURYL CHLORIDE is selected from: methylsulfonyl chloride, benzene sulfonyl chloride, substituted phenylsulfonyl chloride (as halogeno-benzene SULPHURYL CHLORIDE, alkylbenzene sulfonyl chloride) etc., more preferably methylsulfonyl chloride.Described temperature of reaction is spent to reflux temperature for-5, and preferably room temperature is to reflux temperature.Described reaction solvent is selected from benzene, toluene, halogenated aryl hydrocarbon, halogenated alkane (as chloroform or methylene dichloride), tetrahydrofuran (THF), acetonitrile and ionic liquid.More preferably, described reaction back flow reaction in methylene dichloride system.
(4) formula V compound and reaction of sodium azide, preferably, in DMF system, react acquisition formula (VI) compound at 60 ℃ of temperature;
(5) formula (VI) compound and ammonium chloride and zinc powder, iron powder or palladium carbon catalytic reduction preparation formula (IV) compound, preferred described catalyzer carries out catalysis under mineral acid condition, is preferably hydrochloric acid or sulfuric acid, and described catalyzer is preferably zinc powder and ammonium chloride.
In step (5), preferred reaction solvent is water or organic solvent (for example alcohols, halogenated hydrocarbon, aromatic hydrocarbons etc.) or its mixture, is preferably the reaction system of second alcohol and water.
Preferred synthetic route is as follows:
More preferably, described preparation method is as shown in following flow process:
Meanwhile, for Z, be other substituent formulas (III) or (IV) compound, for example R
5for other substituent-NR
5, C (R
6)
2, during S, can adopt corresponding proyl chlorine, the preparation of propine mercaptan.
According to the present invention, 5 shown in described formula II, the 6-chloro-thiophene of bis-replacement-4-[2,3-d] pyrimidine is prepared by following process:
(1) by 3-amino 4,5-bis-substituted thiophenes-2-methyl-formiate (VII) and FORMAMIDINE ACETATE form the dibasic Thienopyrimidine ketone compound of 5,6-(VIII) under the effect of reagent 1, preferably, described temperature of reaction is 0 to spend to reflux temperature, and more preferably room temperature is to reflux temperature;
(2) compound (VIII) forms 5 shown in formula II under reagent 2 effect, the 6-chloro-thiophene of bis-replacement-4-[2,3-d] pyrimidine, and preferably, described temperature of reaction is 0 to spend to reflux temperature, more preferably room temperature is to reflux temperature.
Concrete visible following reaction formula
Wherein, Reagent 1 is selected from: benzene, toluene, ethylene glycol monomethyl ether, glycol dimethyl ether, ethylene glycol monoethyl ether, ethylene glycol diethyl ether, ethanol, 1,2-ethylene dichloride and ionic liquid etc.; Reagent 2 is selected from: thionyl chloride, and phosphorus oxychloride, phosphorus pentachloride chlorine replaces reagent etc.
According to the present invention, the preparation method of thiophene [2, the 3-d] pyrimidine compound of the Han isoxazole heterocycle shown in a kind of formula (I) is provided, it is characterized in that, described method comprises the steps:
(1) substituted benzaldehyde and azanol or hydroxylamine hydrochloride are in methanol/water system, and under the catalysis of sodium carbonate, preferably room temperature is reacted the corresponding benzaldoxime of generation to reflux temperature;
(2) benzaldoxime step (1) being made and propiolic alcohol form formula (III) isoxazole compound by 1,3-Dipolar Cycloaddition under the effect of N-bromo-succinimide (NCS) and triethylamine;
(3) (III) isoxazole compound reacts with methylsulfonyl chloride formula, preferably in methylene dichloride system, obtain formula V compound under room temperature;
(4) formula V compound and reaction of sodium azide, preferably, in DMF system, react acquisition formula (VI) compound at 60 ℃ of temperature;
(5) formula (VI) compound is under zinc powder and ammonium chloride catalysis, the preparation formula that refluxes in ethanol/water system (IV) compound;
(6) 3-amino 4, and 5-bis-substituted thiophenes-2-methyl-formiate (VII) and FORMAMIDINE ACETATE form the dibasic Thienopyrimidine ketone compound of 5,6-(VIII) under the effect of reagent 1;
(7) compound (VIII) forms 5 shown in formula II under reagent 2 effects, the 6-chloro-thiophene of bis-replacement-4-[2,3-d] pyrimidine,
(8) shown in formula II 5, the chloro-thiophene [2 of 6-bis-replacement-4-, 3-d] 3-substituted-phenyl-5-methylol-isoxazoles shown in pyrimidine and formula III or 3-substituted-phenyl-5-aminomethyl-isoxazoles shown in formula IV, in dry organic solvent and alkaline acid binding agent system, reaction prepares the compound shown in formula (I-1) or formula (I-2).
If needed, can form pharmacy acceptable salt or the solvate of formula (I) compound.
According to the present invention, described formula I compound, includes but not limited to: their optical isomer, racemic modification and composition thereof.
According to the present invention, described halogen or halogen atom are selected from fluorine, chlorine or bromine.
According to the present invention, described alkyl is straight or branched alkyl, as C
1-6alkyl, preferably C
1-3alkyl, includes but not limited to methyl, ethyl, propyl group, sec.-propyl, butyl, isobutyl-, the tertiary butyl, n-pentyl, isopentyl, neo-pentyl, n-hexyl etc.
According to the present invention, 4 to 6 yuan of described carbocyclic rings are selected from cyclobutyl, cyclopentyl, cyclohexyl.Be preferably cyclopentyl or cyclohexyl.
According to the present invention, described contain at least one and be selected from N, O, the heteroatomic heterocycle of S is selected from tetrahydrofuran (THF), tetramethylene sulfide, azetidine, pyrrolidyl, acridine, Pyrrolidine, 1,3-thiazoline, 1,3-dihydro-oxazole, piperidines, piperazine, morpholine, thiomorpholine, thiazine, is preferably piperazinyl, morpholinyl or piperidyl.
According to the present invention, described aryl is monocycle or Bicyclic alkyl, for example C
6-14aryl, described aryl is preferably phenyl or naphthyl, more preferably phenyl.
According to the present invention, described heteroaryl is the assorted aryl radical of monocycle or dicyclo, wherein containing heteroatomic ring, be preferably 5 Yuans or 6 Yuans heteroaryls, it contains at least one, preferably have 1-4 N, the heteroatomic aromatic nucleus of S or O, described heteroaryl is preferably pyridyl, pyrimidyl, quinolyl, isoquinolyl, quinazolyl, quinoxalinyl, indyl, pyrryl, thienyl, furyl, benzofuryl, benzothienyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrazolyl, furazan base, thiadiazolyl group, tetrazyl etc., pyrryl more preferably, thienyl, furyl, indyl, benzofuryl.
Term " significant quantity " refers to: described at least one compound and/or at least one pharmacy acceptable salt for can be effectively " treatment ' individual a kind of disease or uncomfortable consumption.If during cancer, significant quantity can reduce the number of cancer or tumour cell; Dwindle the size of tumour; Suppress or stop tumour cell to the intrusion of periphery organ.For example: tumour spreads in soft tissue or bone; Suppress or stop the transfer of tumour; Suppress or stop the growth of tumour; Alleviate to a certain extent one or more symptoms relevant to cancer; Reduce M & M; Improve the quality of living; Or the combination of above-mentioned effect.Significant quantity can be by suppressing EGFR activity, to reduce the consumption of disease symptoms.For cancer therapy, in body, the effect of experiment can be by assessment as survival time, progression of disease time (Time to Disease Progression, TTP), reactivity (response Rates, RR) sustained reaction phase and/or quality of life are measured.Professional recognizes, significant quantity can be along with the approach of administration, the consumption of vehicle and changing with share of other drug.
Term " significant quantity " also can refer to such an extent that be described at least one compound and/or overexpression and/or the hyperactivity effective dosage of its at least one pharmacy acceptable salt to inhibition EGFR.
Compound of the present invention all has antitumor, antitumour activity, especially has Chinese People's Anti-Japanese Military and Political College colon-cancer cell pearl (HCT-116) activity and anti-human lung cell A549 active.Compound of the present invention is 1 * 10
-4under M concentration, all have and suppress active, the activity of embodiment compound can reach more than 50%, preferably more than 60%, more preferably more than 80%.Wherein most compounds has stronger inhibition activity to colorectal cancer cells pearl (HCT-116) and human lung cancer cell A549 simultaneously.For example compound S-101 are 1 * 10
-4under M concentration, to the inhibiting rate of colorectal cancer cells pearl (HCT-116), be 68.8%, and be 88% to human lung cancer cell A549's inhibiting rate, compound S-3 are 1 * 10
-4under M concentration, to the inhibiting rate of colorectal cancer cells pearl (HCT-116), be 70%, and be 86.4% to human lung cancer cell A549's inhibiting rate, compound S-83 are 1 * 10
-4under M concentration to human lung cancer cell A549's inhibiting rate up to 89.4%.Thereby compound of the present invention can be used as treating drug candidate or the lead compound of tumour, cancer.
Concrete embodiment
Below in conjunction with embodiment, the present invention is further illustrated.It should be noted that, embodiment compound is not limiting the scope of the invention, and any improvement of making on basis of the present invention and variation are all without prejudice to spirit of the present invention.
Wherein, the building-up process of intermediate and target compound is all with the representative explanation in embodiment, the same representation compound of building-up process of remaining intermediate and target compound.
Instrument and reagent
AVANCE III nuclear magnetic resonance analyser (400MHz, DMSO-d
6tMS is interior mark), ion trap liquid matter is used in conjunction instrument (DECAX-30000LCQ Deca XP), Shimadzu FTIR-8400S (Japanese Shimadzu company produce), XT5 numeral shows that (Beijing tech electric light instrument plant manufactures micro melting point apparatus, temperature is not calibrated), the wavelengthtunable orifice plate microplate reader (Molecular Devies SPECTRAMAX190) that declines.Chemical reagent is commercially available analytical pure or chemically pure reagent, RPMI1640 is purchased from Gibco company, sulphonyl rhodamine B (SulforodamineB, SRB) is purchased from Sigma company, and trichoroacetic acid(TCA) (TCA), acetic acid and Tris base unbuffer are domestic analytical reagent.
Synthesizing of embodiment 1 formula (III) intermediate 3-substituted-phenyl-5-methylol-isoxazoles and formula (IV) intermediate 3-substituted-phenyl 5-aminomethyl-isoxazoles
Wherein with R
3for H is as example:
(1) benzaldoxime is synthetic
10.0mmol dissolution of benzaldehyde is at 30mL 30%CH
3oH and H
2in O solution, add in the triangular flask that magnetic stirring apparatus is housed, under stirring, add 10.0mmol oxammonium hydrochloride, slowly add the sodium carbonate of the dry porphyrize of 5.0mmol after waiting oxammonium hydrochloride to dissolve.Room temperature reaction, after TLC detection reaction completes, system removes under reduced pressure after methyl alcohol, adds 30mL H
2o, methylene dichloride (3 * 30mL) extraction, merges organic layer, anhydrous sodium sulfate drying organic layer. and desolventizing obtains benzaldoxime crude product, yield 86.2%.This crude product directly carries out the next step without separation and purification.
(2) 3-phenyl-5-methylol-isoxazoles
10.0mmol benzaldoxime and the dry methylene dichloride of 30mL are added in 250mL single necked round bottom flask, under stirring, add 1.60g (12.0mmol) N-neoprene imide (NCS), slightly be heated to after the whole dissolvings of NCS, drip 0.56g (10.0mmol) 2-propine-1-alcohol, slowly be added dropwise to subsequently 20mL containing the dichloromethane solution of 10.1g (10.0mmol) triethylamine, after adding rear system backflow .TLC detection reaction and completing, mother liquor washing, anhydrous sodium sulfate drying, the separated (V of post
sherwood oil: V
ethyl acetate5:1-2:1) obtain 3-phenyl-5-methylol-isoxazoles, yield 76.8%.
(3) 3-phenyl-5-aminomethyl-isoxazoles
10.0mmol 5-methylol-3-phenyl-isoxazoles and the dry methylene dichloride of 30mL are added in 250mL single necked round bottom flask, ice bath adds the 20mL dichloromethane solution containing 1.01g (10.0mmol) triethylamine in system under stirring, then the 5mL dichloromethane solution that is dissolved with 1.37g (12.0mmol) methylsulfonyl chloride (MsCl) is slowly added dropwise in system, after ice bath reaction 2h, room temperature reaction.After TLC detection reaction completes, mother liquor washing, 5% sodium hydrogen carbonate solution is washed, washing, anhydrous sodium sulfate drying, decompression desolventizing obtains crude product 5-methylsulfonic acid-3-phenyl-isoxazole-5-methyl alcohol ester, yield 68.0%.Crude product needn't directly carry out the next step by purifying.
5.0mmol 5-methylsulfonic acid-3-phenyl-isoxazoles-5-methyl alcohol ester is dissolved in the DMF that 20mL is dry, add 0.34g (5.20mmol) sodiumazide, after stirring at room is dissolved, being placed in 45 ℃ of-50 ℃ of oil baths reacts, after TLC Indicator Reaction completes, filter, ether for filter cake (2 * 30mL) washing, merge organic layer, in organic layer, add 100mL water, with ether (5 * 30mL), extract, merge organic layer, organic layer washing 2 times, anhydrous sodium sulfate drying, after desolventizing, obtain crude product 3-phenyl-5-azido-methyl-isoxazoles, yield 90%, crude product directly carries out reduction reaction below.
5.0mmol 5-azido-methyl-3-phenyl-isoxazoles are dissolved in the mixing solutions of 80mL ethanol and 20mL water, by the NH of the zinc powder of 0.17g (2.6mmol) and 0.28g (5.2mmol)
4cl adds system, backflow 1h, and vacuum is sloughed ethanol, to system, add 20mL water, the sodium hydroxide solution with 20% is adjusted to pH value to 12, in system, adds 50mL DCM, after stirring, filter, filter residue is in the water dissolution with a small amount of, filter, merge the filtrate of twice, separate organic layer, organic layer, washing, anhydrous sodium sulfate drying, vacuum desolvation agent, the separated (V of residue post
methylene dichloride: V
methyl alcohol, 10:1) obtain product 3-phenyl-5-aminomethyl-isoxazoles, yield 75%. light yellow solids, m.p:39-40 ℃,
1h-NMR (400MHz, CDCl
3, TMS), δ ppm:1.60(s, 2H, NH
2), 3.91 (s, 2H, CH
2), 6.40 (s, 1H), 7.39 (m, 2H, Ar-H), 7.76 (m, 2H, Ar-H).
Embodiment 2
Synthetic (with the routine explanation of synthesizing of the chloro-thiophene of the 6-tertiary butyl-4-[2,3-d] pyrimidine) of intermediate (II):
2.13g (10mmol) 3-amino-5-tertiary butyl-thiophene-2-carboxylic acid methyl esters is added in 250mL round bottom single necked round bottom flask, then add 100mL ethylene glycol monoethyl ether and 2.14g (20mmol) FORMAMIDINE ACETATE, system is back flow reaction under agitation, after TLC detection reaction completes, system is cooling, then underpressure distillation to ethylene glycol monoethyl ether has 30mL, system refrigerates a few hours, there is a large amount of solids to separate out, filter, the ether washing several of refrigeration for filter cake, is directly used in the next step after filter cake vacuum-drying.
Taking 5 grams of above-mentioned crude products adds in 100mL round bottom single necked round bottom flask, slowly drip the phosphorus oxychloride solution 20mL of 3 DMF, after system back flow reaction 6h, remove under reduced pressure after excessive phosphorus oxychloride, mix silica gel after cooling, post separated (V (sherwood oil): V (ethyl acetate)=4:1-1:1) obtains the chloro-thiophene [2 of the target compound 6-tertiary butyl-4-, 3-d] pyrimidine, light yellow solid, 1H-NMR (400MHz, CDCl3, TMS), δ ppm:1.43(s, 9H, 3CH3), 7.19 (s, 1H), 8.84 (s, 1H).
Embodiment 3
5-methyl-4-[(3-phenyl-isoxazole-5-bases)-methoxyl group-]-thiophene [2,3-d] pyrimidine synthetic
By the chloro-thiophene [2 of the 4-of 0.184g (1mmol), 3-d] pyrimidine is dissolved in the Virahol that 5mL is dry, under stirring, the 5mL aqueous isopropanol that is dissolved with 0.175g (1mmol) 5-methylol-3-phenyl-isoxazoles is slowly added dropwise to reaction system, then add the new triethylamine steaming of 0.101g (1mmol), after system stirring at room 30min, 60 ℃ of reactions, after TLC detection reaction completes, reaction solution vacuum concentration, the separated V of the direct post of residue
(sherwood oil): V
(ethyl acetate)=9:1-4:1) obtain target compound 5-methyl-4-[(3-phenyl-isoxazole-5-bases)-methoxyl group-]-thiophene [2,3-d] pyrimidine (following table S-1).Remaining compound is according to 5-methyl-4-[(3-phenyl-isoxazole-5-bases)-methoxyl group-] building-up process of-thiophene [2,3-d] pyrimidine is synthetic.Its structure is all by IR,
1h NMR, the analytical procedures such as ESI-MS characterize.The Physical Constants of the compound optimizing and spectroscopic data describe with the form of list:
The structure of compound, code name and MS data are as shown in the table:
The preferred compound of table 1-
States of matter and the IR data of table 2-table 1 compound
| Numbering | States of matter | IR/cm -1 |
| S-1 | White solid | 3103,1567,1546,1442,1309,1026,770 |
| S-2 | White solid | 3107,1567,1548,1457,1444,1312,1028,817 |
| S-3 | White solid | 3107,1567,1548,1457,1444,1312,1028,821 |
| S-4 | White solid | 3107,1567,1548,1457,1444,1312,1028,836 |
| S-5 | White solid | 3107,1567,1548,1457,1444,1312,1028,829 |
| S-6 | White solid | 3107,1567,1548,1457,1444,1312,1028,817 |
| S-7 | White solid | 3107,1567,1548,1457,1444,1312,1028,845 |
| S-8 | White solid | 3107,1567,1548,1457,1444,1312,1028,847 |
| S-9 | White solid | 3107,1567,1548,1457,1444,1312,1028,817 |
| S-10 | White solid | 3107,1567,1548,1457,1444,1312,1028,816 |
| S-11 | White solid | 3237,3103,1567,1546,1442,1309,1026,770 |
| S-12 | White solid | 3230,3107,1567,1548,1457,1444,1312,1028,817 |
| S-13 | White solid | 3368,3107,1567,1548,1457,1444,1312,1028,821 |
| S-14 | White solid | 3228,3107,1567,1548,1457,1444,1312,1028,836 |
| S-15 | White solid | 3231,3107,1567,1548,1457,1444,1312,1028,829 |
| S-16 | White solid | 3238,3107,1567,1548,1457,1444,1312,1028,817 |
| S-17 | White solid | 3210,3107,1567,1548,1457,1444,1312,1028,845 |
| S-18 | White solid | 3218,3107,1567,1548,1457,1444,1312,1028,847 |
| S-19 | White solid | 3228,3107,1567,1548,1457,1444,1312,1028,817 |
| S-20 | White solid | 3218,3107,1567,1548,1457,1444,1312,1028,816 |
| S-21 | White solid | 3107,1567,1548,1457,1444,1312,1028,756 |
| S-22 | White solid | 3105,1567,1545,1457,1441,1312,1028,817 |
| S-23 | White solid | 3107,1567,1545,1457,1440,1312,1028,820 |
| S-24 | White solid | 3107,1565,1548,1457,1444,1312,1028,815 |
| S-25 | White solid | 3107,1567,1548,1457,1444,1312,1028,830 |
| S-26 | White solid | 3107,1566,1548,1456,1443,1312,1028,835 |
| S-27 | White solid | 3107,1567,1548,1457,1444,1312,1028,817 |
| S-28 | White solid | 3107,1567,1548,1457,1444,1312,1028,817 |
| S-29 | White solid | 3107,1567,1548,1457,1444,1312,1028,817 |
| S-30 | White solid | 3107,1567,1548,1457,1444,1312,1028,817 |
| S-31 | White solid | 3237,3103,1567,1546,1442,1309,1026,770 |
| S-32 | White solid | 3237,3103,1567,1546,1442,1309,1026,770 |
| S-33 | White solid | 3237,3103,1567,1546,1442,1309,1026,770 |
| S-34 | White solid | 3237,3103,1567,1546,1442,1309,1026,770 |
| S-35 | White solid | 3237,3103,1567,1546,1442,1309,1026,770 |
| S-36 | White solid | 3237,3103,1567,1546,1442,1309,1026,770 |
| S-37 | White solid | 3237,3103,1567,1546,1442,1309,1026,770 |
| S-38 | White solid | 3237,3103,1567,1546,1442,1309,1026,770 |
| S-39 | White solid | 3237,3103,1567,1546,1442,1309,1026,770 |
| S-40 | White solid | 3237,3103,1567,1546,1442,1309,1026,770 |
| S-41 | White solid | 3103,1567,1546,1442,1309,1026,770 |
| S-42 | White solid | 3103,1567,1546,1442,1309,1026,812 |
| S-43 | White solid | 3103,1567,1546,1442,1309,1026,835 |
| S-44 | White solid | 3103,1567,1546,1442,1309,1026,850 |
| S-45 | White solid | 3103,1567,1546,1442,1309,1026,832 |
| S-46 | White solid | 3103,1567,1546,1442,1309,1026,867 |
| S-47 | White solid | 3103,1567,1546,1442,1309,1026,856 |
| S-48 | White solid | 3103,1567,1546,1442,1309,1026,834 |
| S-49 | White solid | 3103,1567,1546,1442,1309,1026,865 |
| S-50 | White solid | 3103,1567,1546,1442,1309,1026,823 |
| S-51 | White solid | 3237,3103,1567,1546,1442,1309,1026,770 |
| S-52 | White solid | 3237,3103,1567,1546,1442,1309,1026,815 |
| S-53 | White solid | 3237,3103,1567,1546,1442,1309,1026,834 |
| S-54 | White solid | 3237,3103,1567,1546,1442,1309,1026,850 |
| S-55 | White solid | 3237,3103,1567,1546,1442,1309,1026,867 |
| S-56 | White solid | 3237,3103,1567,1546,1442,1309,1026,834 |
| S-57 | White solid | 3237,3103,1567,1546,1442,1309,1026,821 |
| S-58 | White solid | 3237,3103,1567,1546,1442,1309,1026,845 |
| S-59 | White solid | 3237,3103,1567,1546,1442,1309,1026,860 |
| S-60 | White solid | 3237,3103,1567,1546,1442,1309,1026,835 |
| S-61 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,775 |
| S-62 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,810 |
| S-63 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,823 |
| S-64 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,835 |
| S-65 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,840 |
| S-66 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,860 |
| S-67 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,851 |
| S-68 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,875 |
| S-69 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,835 |
| S-70 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,832 |
| S-71 | White solid | 3237,3103,1567,1546,1442,1309,1026,775 |
| S-72 | White solid | 3237,3103,1567,1546,1442,1309,1026,821 |
| S-73 | White solid | 3237,3103,1567,1546,1442,1309,1026,835 |
| S-74 | White solid | 3237,3103,1567,1546,1442,1309,1026,845 |
| S-75 | White solid | 3237,3103,1567,1546,1442,1309,1026,835 |
| S-76 | White solid | 3237,3103,1567,1546,1442,1309,1026,851 |
| S-77 | White solid | 3237,3103,1567,1546,1442,1309,1026,823 |
| S-78 | White solid | 3237,3103,1567,1546,1442,1309,1026,853 |
| S-79 | White solid | 3237,3103,1567,1546,1442,1309,1026,835 |
| S-80 | White solid | 3237,3103,1567,1546,1442,1309,1026,843 |
| S-81 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,775 |
| S-82 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,845 |
| S-83 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,845 |
| S-84 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,845 |
| S-85 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,845 |
| S-86 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,845 |
| S-87 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,845 |
| S-88 | White solid | 3111,1571,1548,1512,1461,1429,1372,1322,1036,846 |
| S-89 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,845 |
| S-90 | White solid | 3109,1571,1549,1510,1461,1431,1363,1322,1036,827 |
| S-91 | White solid | 3237,3109,1571,1549,1461,1431,1363,1322,1036,775 |
| S-92 | White solid | 3230,3109,1571,1549,1461,1431,1363,1322,1036,845 |
| S-93 | White solid | 3368,3109,1571,1549,1461,1431,1363,1322,1036,845 |
| S-94 | White solid | 3228,3109,1571,1549,1461,1431,1363,1322,1036,845 |
| S-95 | White solid | 3231,3109,1571,1549,1461,1431,1363,1322,1036,845 |
| S-96 | White solid | 3238,3109,1571,1549,1461,1431,1363,1322,1036,845 |
| S-97 | White solid | 3210,3109,1571,1549,1461,1431,1363,1322,1036,845 |
| S-98 | White solid | 3218,3111,1571,1548,1461,1429,1372,1322,1036,,846 |
| S-99 | White solid | 3228,3109,1571,1549,1461,1431,1363,1322,1036,845 |
| S-100 | White solid | 3218,3109,1571,1549,1461,1431,1363,1322,1036,827 |
| S-101 | White solid | 3103,1567,1546,1442,1309,1026,770 |
| S-102 | White solid | 3107,1567,1548,1457,1444,1312,1028,817 |
| S-103 | White solid | 3107,1567,1548,1457,1444,1312,1028,821 |
| S-104 | White solid | 3107,1567,1548,1457,1444,1312,1028,836 |
| S-105 | White solid | 3107,1567,1548,1457,1444,1312,1028,829 |
| S-106 | White solid | 3107,1567,1548,1457,1444,1312,1028,817 |
| S-107 | White solid | 3107,1567,1548,1457,1444,1312,1028,845 |
| S-108 | White solid | 3107,1567,1548,1457,1444,1312,1028,847 |
| S-109 | White solid | 3107,1567,1548,1457,1444,1312,1028,817 |
| S-110 | White solid | 3107,1567,1548,1457,1444,1312,1028,816 |
| S-111 | White solid | 3237,3103,1567,1546,1442,1309,1026,770 |
| S-112 | White solid | 3230,3107,1567,1548,1457,1444,1312,1028,817 |
| S-113 | White solid | 3368,3107,1567,1548,1457,1444,1312,1028,821 |
| S-114 | White solid | 3228,3107,1567,1548,1457,1444,1312,1028,836 |
| S-115 | White solid | 3231,3107,1567,1548,1457,1444,1312,1028,829 |
| S-116 | White solid | 3238,3107,1567,1548,1457,1444,1312,1028,817 |
| S-117 | White solid | 3210,3107,1567,1548,1457,1444,1312,1028,845 |
| S-118 | White solid | 3218,3107,1567,1548,1457,1444,1312,1028,847 |
| S-119 | White solid | 3228,3107,1567,1548,1457,1444,1312,1028,817 |
| S-120 | White solid | 3218,3107,1567,1548,1457,1444,1312,1028,816 |
| S-121 | White solid | 3109,1561,1531,1510,1455,1434,1386,1311,1028,826 |
| S-122 | White solid | 3124,2957,1613,1571,1562,1460,1396,1311,1075,1058,888 |
| S-123 | White solid | 3124,2957,1613,1571,1562,1460,1396,1311,1075,1058,888 |
| S-124 | White solid | 3124,2957,1613,1571,1562,1460,1432,1311,1075,1058,888 |
| S-125 | White solid | 3124,2957,1613,1571,1562,1460,1432,1311,1075,1058,888 |
| S-126 | White solid | 3124,2957,1613,1571,1562,1460,1396,1311,1075,1058,888 |
| S-127 | White solid | 3124,2957,1613,1571,1562,1460,1396,1311,1075,1058,888 |
| S-128 | White solid | 3124,2957,1613,1571,1562,1460,1432,1311,1075,1058,888 |
| S-129 | White solid | 3124,2957,1613,1571,1562,1460,1432,1311,1075,1058,888 |
| S-130 | White solid | 3124,2957,1613,1571,1562,1460,1432,1311,1075,1058,888 |
| S-131 | White solid | 3237,3109,1561,1531,1510,1455,1434,1386,1028,826 |
| S-132 | White solid | 3230,3124,2957,1613,1571,1562,1460,1396,1311,1075,888 |
| S-133 | White solid | 3368,3124,2957,1613,1571,1562,1460,1432,1396,888 |
| S-134 | White solid | 3228,3124,2957,1613,1571,1562,1460,1432,1396,888 |
| S-135 | White solid | 3231,3124,2957,1613,1571,1562,1460,1432,1396,1075 |
| S-136 | White solid | 3238,3124,2957,1613,1571,1562,1460,1432,1396,1075 |
| S-137 | White solid | 3210,3124,2957,1613,1571,1562,1460,1432,1396,1311 |
| S-138 | White solid | 3218,3124,2957,1613,1571,1562,1460,1432,1396,1075 |
| S-139 | White solid | 3228,3124,2957,1613,1571,1562,1460,1432,1396,1311,1057 |
| S-140 | White solid | 3218,3124,2957,1613,1571,1562,1460,1432,1396,1311 |
Compound in table 3-table 1
1hNMR data
The preparation of embodiment 4 the compounds of this invention pharmacy acceptable salts
With 5-methyl-4-[(3-phenyl-isoxazole-5-base)-methoxyl group-] hydrochloride of-thiophene [2,3-d] pyrimidine and the preparation of acetate illustrate it, the preparation of the salt of all the other compounds is with this process.
(1) 5-methyl-4-[(3-phenyl-isoxazole-5-bases)-methoxyl group-] preparation of-thiophene [2,3-d] pyrimidine hydrochloride
By 5-methyl-4-[(3-phenyl-isoxazole-5-bases of 0.5mmol)-methoxyl group-]-thiophene [2,3-d] pyrimidine adds 20mL (V:V, 1:1) in 5% hydrochloric acid soln and methanol mixed solution, after low-grade fever stirs it is dissolved, the crystallization of room temperature slow evaporation obtains 5-methyl-4-[(3-phenyl-isoxazole-5-bases)-methoxyl group-]-thiophene [2,3-d] pyrimidine hydrochloride, white solid, yield: 72%.
(2) 5-methyl-4-[(3-phenyl-isoxazole-5-bases)-methoxyl group-] preparation of-thiophene [2,3-d] pyrimidine acetate:
By 0.5mmol 5-methyl-4-[(3-phenyl-isoxazole-5-bases)-methoxyl group-]-thiophene [2,3-d] pyrimidine adds in the 50mL single necked round bottom flask that fills 10mL dry methylene chloride, under stirring, add 2mL glacial acetic acid, 30 ℃-40 ℃ are stirred 1-2h, and cooling rear refrigeration crystallization is filtered, after vacuum-drying, obtain 5-methyl-4-[(3-phenyl-isoxazole-5-bases)-methoxyl group-]-thiophene [2,3-d] pyrimidine acetate, colorless solid, yield: 58%.
Embodiment 5
Adopt srb assay to carry out anti-colorectal cancer cell lines (HCT-116) and human lung carcinoma cell line (A549) screening active ingredients, screening process reference (Li M.H.; Miao Z.H.; Tan W.F.et al.Clin.Cancer Res.2004,10 (24): 8266-8274).
Concrete experimental procedure is as follows:
(1) according to the growth velocity of tumour cell, the colorectal cancer cell lines in logarithmic phase (HCT-116) is inoculated in to 96 well culture plates, adherent growth is after 24 hours, by 1 * 10
-4the medicine of M adds, and its concentration is established 3 multiple holes, and establishes physiological saline contrast and the acellular zeroing hole of respective concentration, and lesion/cancer cell is cultivated 72 hours under 37 ℃, 5%CO2 condition; (2) take out culture plate, 10% cold nitrilotriacetic (TCA) solution fixed cell for every hole, places 1 hour for 4 ℃; (3) abandon stationary liquid, use distilled water wash 5 times, seasoning in air; (4) then add the SRB solution of being prepared by 1% glacial acetic acid, room temperature dyeing 15 minutes; (5) remove supernatant liquor, the acetum washing with 1% 5 times, dry air; (6) finally add Tris solution, on dull and stereotyped oscillator, shake 5 minutes.Absorbance (the A declining under orifice plate microplate reader survey 560nm wavelength with wavelengthtunable
560).Adopt following formula to calculate enzyme inhibition rate.
Record the compound or its salt shown in formula (I) 1 * 10
-4the activity that suppresses colorectal cancer cells pearl (HCT-116) and human lung cancer cell A549 under M concentration.Experimental result sees the following form 5 and table 6.
In table 5-formula (I), part embodiment compound suppresses colorectal cancer cells pearl (HCT-116) active testing result
| Compound number | Inhibiting rate (%) | Compound number | Inhibiting rate (%) |
| S-3 | 70.0 | S-101 | 68.8 |
| S-83 | 69.6 | S-102 | 62.9 |
| S-87 | 72.7 | S-103 | 71.1 |
| S-88 | 60.9 | S-107 | 72.9 |
In table 6-formula (I), part embodiment compound suppresses human lung cancer cell A549's active testing result
| Compound number | Inhibiting rate (%) | Compound number | Inhibiting rate (%) |
| S-1 | 85.0 | S-8 | 85.9 |
| S-2 | 80.0 | S-83 | 89.4 |
| S-3 | 86.4 | S-101 | 88.0 |
| S-5 | 80.8 | S-106 | 71.6 |
| S-6 | 75.6 | S-121 | 54.1 |
Claims (10)
1. thiophene [2,3-d] pyrimidine derivatives or its pharmacy acceptable salt or the solvate shown in a formula (I),
Wherein:
R
1and R
2can be identical or different, be selected from independently of one another hydrogen, C
1-6alkyl, C
1-6alkoxyl group, halo C
1-6alkyl, halo C
1-6the C that alkoxyl group or hydroxyl replace
1-6alkyl, optionally by R
7the aryl replacing or optionally by R
8the heteroaryl replacing;
Or R
1and R
2coupled carbon atom forms 4 to 6 yuan of carbocyclic rings or heterocycles together, and described carbocyclic ring or heterocycle are optionally by R
11replace; Described heterocycle contains at least 1 and is selected from N, the heteroatoms of O or S;
Z is-NR
5-, C (R
6)
2,-S-or-O-, wherein R
5for hydrogen or C
1-6alkyl, R
6identical or different, be selected from hydrogen, C
1-6alkyl, halo C
1-6alkyl or the C that replaces of hydroxyl
1-6alkyl;
R
3be selected from hydrogen, halogen, C
1-6alkyl, C
1-6alkoxyl group, halo C
1-6alkyl, halo C
1-6the C that alkoxyl group or hydroxyl replace
1-6alkyl, the integer that n is 0 ~ 5;
R
4be selected from hydrogen, C
1-6alkyl, C
1-6alkoxyl group, halo C
1-6alkyl or halo C
1-6alkoxyl group, optionally by R
9the aryl replacing or optionally by R
10the heteroaryl replacing;
R
7, R
8, R
9, R
10or R
11be selected from independently of one another hydrogen, hydroxyl, sulfydryl, cyano group, amino, C
1-6alkylamino, two (C
1-6alkyl) amino, nitro, halogen, C
1-6alkyl, C
1-6alkoxyl group, C
1-6alkylthio, carboxyl, halo C
1-6alkyl, halo C
1-6alkoxyl group or for C
1-6alkylthio.
2. according to the thiophene of claim 1 [2,3-d] pyrimidine derivatives or its pharmacy acceptable salt or solvate, wherein
R
1and R
2can be identical or different, be selected from independently of one another hydrogen, C
1-3alkyl, C
1-6alkoxyl group, halo C
1-6alkyl, halo C
1-6alkoxyl group, phenyl or by R
7the phenyl replacing, or
R
1and R
2coupled carbon atom forms 4 to 6 yuan of carbocyclic rings together, and described carbocyclic ring is optionally by hydrogen, C
1-6alkyl, halogen, nitro or amino replacement;
R
4be selected from hydrogen, C
1-6alkyl, C
1-6alkoxyl group, halo C
1-6alkyl, phenyl or by R
9the phenyl replacing.
3. according to the thiophene of claim 1 or 2 [2,3-d] pyrimidine derivatives or its pharmacy acceptable salt or solvate, wherein
R
1and R
2be selected from independently of one another hydrogen, C
1-3alkyl, phenyl or by R
7the phenyl replacing, more preferably hydrogen, methyl, ethyl, propyl group, sec.-propyl, butyl, isobutyl-, the tertiary butyl or phenyl;
Z is-NH-, CH
2or-O-;
R
3be selected from hydrogen, fluorine, chlorine, bromine, methyl, methoxyl group or trifluoromethyl, n is preferably 1-4, more preferably 2-3.
R
4be selected from hydrogen, C
1-6alkyl, C
1-6alkoxyl group or halo C
1-6alkyl, phenyl or substituted-phenyl.
Also more preferably, R
1and R
2can be identical or different, be selected from independently of one another C
1-6alkyl, phenyl or by R
7the phenyl replacing, more preferably hydrogen, methyl, ethyl, the tertiary butyl, phenyl, more preferably R
1be selected from hydrogen, methyl, phenyl; R
2be selected from hydrogen, methyl, ethyl or the tertiary butyl;
Z is-NH-or-O-, R
3preferably ortho position or the contraposition of isoxazole ring, more preferably 4-fluorine, 4-chlorine, 2-chlorine, 4-bromine, 2,4-dichloro, 4-methyl, 4-methoxyl group, hydrogen, 4-trifluoromethyl or 2,4-dimethoxy;
R
4be selected from hydrogen, methyl or phenyl.
4. according to the thiophene of claim 1-3 any one [2,3-d] pyrimidine derivatives or its pharmacy acceptable salt, wherein said compound is selected from:
5. a pharmaceutical composition, it comprises the thiophene [2 shown in claim 1-4 any one formula (I), 3-d] pyrimidine compound, or its pharmacy acceptable salt, and at least one pharmaceutically acceptable, inertia, nontoxic vehicle or carrier or thinner.Preferably, described pharmaceutical composition also comprises the acceptable subsidiary material of one or more pharmacy that are selected from weighting agent, disintegrating agent, lubricant, glidant, effervescent, correctives, sanitas and coating material.
6. pharmaceutical composition according to claim 5, is characterized in that, described pharmaceutical composition is dosage form, is preferably solid orally ingestible, liquid oral medicine or injection.Preferably, described preparation is selected from tablet, dispersible tablet, enteric coated tablet, chewable tablet, orally disintegrating tablet, capsule, granule, oral solution, injection liquid drugs injection, injection freeze-dried powder, infusion solutions or primary infusion.
7. as thiophene [2,3-d] pyrimidine compound or its pharmacy acceptable salt shown in the formula (I) of the claim 1-4 of medicine, especially a kind of medicine that is used for the treatment of lesion/cancer disease.
8. thiophene [2, the 3-d] pyrimidine compound shown in the formula (I) of a claim 1-4 any one or its pharmacy acceptable salt are in the application for the preparation of in antitumor or cancer drug.Preferably described tumour or cancer is selected from: bladder cancer, nonsmall-cell lung cancer, ovarian cancer, mammary cancer, cancer of the stomach, esophagus cancer, lung cancer, head and neck cancer, colorectal carcinoma, pharynx cancer and carcinoma of the pancreas etc., more preferably nonsmall-cell lung cancer.
9. in a claim 1-4, thiophene [2, the 3-d] pyrimidine compound shown in the formula (I) of any one and/or pharmacy acceptable salt suppress the application in the transition expression of EGFR and/or the inhibitor of hyperactivity in preparation.
10. the preparation method of thiophene [2, the 3-d] pyrimidine compound shown in the formula (I) of a claim 1-4 any one, is characterized in that: described method comprises the steps:
2,5, the 6-chloro-thiophene of tri-replacement-4-[2,3-d] pyrimidine (formula II) and 3-substituted-phenyl-5-methylol-isoxazoles (formula III) or 3-substituted-phenyl-5-aminomethyl-isoxazoles (formula VI) are raw material, reaction preparation in dry organic solvent and alkaline acid binding agent system:
If needed, form pharmacy acceptable salt or the solvate of formula I compound.
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| EP2835372A1 (en) * | 2012-09-19 | 2015-02-11 | Fujian Institute Of Research On The Structure Of Matter, Chinese Academy Of Sciences | Thiophene [2, 3-d]pyrimidine derivative, and preparation method and use thereof |
| CN105037385A (en) * | 2015-08-18 | 2015-11-11 | 沈阳药科大学 | Piperazine-structure-containing tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidine compounds and application thereof |
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