CN104817493A - Aromatic heterocyclic amide substituted diarylurea compound, preparation method and application thereof - Google Patents
Aromatic heterocyclic amide substituted diarylurea compound, preparation method and application thereof Download PDFInfo
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- CN104817493A CN104817493A CN201510107596.XA CN201510107596A CN104817493A CN 104817493 A CN104817493 A CN 104817493A CN 201510107596 A CN201510107596 A CN 201510107596A CN 104817493 A CN104817493 A CN 104817493A
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- QBDJFVUTUGUABP-UHFFFAOYSA-N CC(C)(C)C(Nc(nc1)ccc1-c(cc1)ccc1NC(Nc1cc(C(F)(F)F)cc(C(F)(F)F)c1)=O)=O Chemical compound CC(C)(C)C(Nc(nc1)ccc1-c(cc1)ccc1NC(Nc1cc(C(F)(F)F)cc(C(F)(F)F)c1)=O)=O QBDJFVUTUGUABP-UHFFFAOYSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/72—Nitrogen atoms
- C07D213/75—Amino or imino radicals, acylated by carboxylic or carbonic acids, or by sulfur or nitrogen analogues thereof, e.g. carbamates
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D213/81—Amides; Imides
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Abstract
The invention provides an aromatic heterocyclic amide substituted diarylurea compound, a preparation method and application thereof. The structural formula of the compound is shown as the specification, wherein Ar is a pyridine heterocycle containing pivaloyl amino or methylamine carbonyl, R1 and R2 are trifluoromethyl, trifluoromethoxy or halogen, R2 can also be alkoxy with a tertiary amine group substituted end and a carbon atom number of 2-3, and the alkoxy is connected to the para-position of urea through the O atom. The compound can be prepared by three-step organic synthesis reaction, and the method has the advantages of easy operation of the reaction process, easily available raw materials, mild reaction conditions, cheap reagents and the like, and is suitable for large-scale production by pharmaceutical enterprises. The compound has good inhibitory activity on VEGFR-2 kinase, also can inhibit the proliferation activity of tumor cells, and can be used for preparation of antitumor drugs and VEGFR-2 kinase activity inhibition drugs, thus having good application prospects and scientific research value.
Description
Technical field
The present invention relates to biomedicine technical field, relate to a kind of antineoplastic compound, be specifically related to di-aryl urea compounds of a kind of fragrant heterocycleamide replacement and its preparation method and application.
Background technology
Malignant tumour as one of larger public health problem in the whole world, the greatly health of harm humans, and will first killer of the new millennium mankind be become.Malignant tumour is no longer the serious disease of advanced industrial country, and developing country is faced with larger Disease Spectrum.
Chemotherapy, as one of important means for the treatment of tumour, has had huge development and progress at nearly 30 years, has obtained large quantities of clinical antitumor agents with different mechanism of action.But antitumour drug also exists many untoward reactions, such as alopecia, vomiting, produces bone marrow depression, and produce resistance etc. fast, these all cause chemicals cannot reach the result for the treatment of of expection.The research and development of therefore new antitumor drug are one of the focus and difficulties of current pharmaceutical field.
Summary of the invention
Di-aryl urea compounds that the object of the present invention is to provide a kind of fragrant heterocycleamide to replace and its preparation method and application, this compound embodies good anti-tumor activity in vitro, can be applied to the preparation of antitumor drug.
For achieving the above object, the technical solution used in the present invention is:
The di-aryl urea compounds that fragrant heterocycleamide replaces, its structural formula is as follows:
Wherein, Ar is the pyridine heterocycle containing pivaloyl amido or methylaminocarbonyl, R
1for trifluoromethyl, trifluoromethoxy or halogen, R
2the carbonatoms replaced by tertiary amine groups for trifluoromethyl, trifluoromethoxy, halogen or end is the alkoxyl group of 2 ~ 3, the carbonatoms that wherein end is replaced by tertiary amine groups be 2 ~ 3 alkoxyl group be connected to the contraposition of urea by O atom.
What be connected with atom N in described tertiary amine groups is straight chained alkyl, cycloalkyl or epoxy alkyl.
Described tertiary amine groups is dimethylamino, morpholinyl or piperidyl.
A preparation method for the di-aryl urea compounds that fragrant heterocycleamide replaces, comprises the following steps:
1) 5-bromo-2-amido pyridine obtains N-(5-bromopyridine-2-base)-2,2-dimethylpropionamide by substitution reaction;
2) N-(5-bromopyridine-2-base)-2,2-dimethylpropionamide and p-aminophenyl borate hydrochlorate obtain N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide by Suzuki linked reaction;
3) N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide obtain by condensation reaction the di-aryl urea compounds that fragrant heterocycleamide replaces with containing disubstituted aniline.(R
1, R
2introduce by containing the substituting group on phenyl ring in disubstituted aniline, R
2can be linked to containing R by Williamson's etherification reaction
1aniline phenyl ring on, thus obtain containing disubstituted aniline)
A preparation method for the di-aryl urea compounds that fragrant heterocycleamide replaces, comprises the following steps:
1) 5-bromine-2-methylpyridine obtains 5-bromopyridine-2-carboxylic acid through oxidizing reaction, and then 5-bromopyridine-2-carboxylic acid is successively through substitution reaction and aminating reaction, the obtained bromo-pyridine carboxamide of N-methyl-5-;
Or 4-bromine-2-methylpyridine obtains 4-bromopyridine-2-carboxylic acid through oxidizing reaction, and then 4-bromopyridine-2-carboxylic acid is successively through substitution reaction and aminating reaction, the obtained bromo-pyridine carboxamide of N-methyl-4-;
2) the bromo-pyridine carboxamide of N-methyl-5-and p-aminophenyl borate hydrochlorate obtain 5-(4-aminophenyl)-N-picoline-2-methane amide by Suzuki linked reaction;
Or the bromo-pyridine carboxamide of N-methyl-4-and p-aminophenyl borate hydrochlorate obtain 4-(4-aminophenyl)-N-picoline-2-methane amide by Suzuki linked reaction;
3) 5-(4-aminophenyl)-N-picoline-2-methane amide obtains by condensation reaction the di-aryl urea compounds that fragrant heterocycleamide replaces with containing disubstituted aniline;
Or 4-(4-aminophenyl)-N-picoline-2-methane amide obtains by condensation reaction the di-aryl urea compounds that fragrant heterocycleamide replaces with containing disubstituted aniline.
The di-aryl urea compounds that virtue heterocycleamide replaces suppresses the application in VEGFR-2 kinase activity medicine in preparation.
The di-aryl urea compounds that virtue heterocycleamide replaces is preparing the application in antitumor drug.
The medicine that described antitumor drug is is target spot with VEGFR-2 kinases, comprises the medicine of anti-lung cancer, anti-liver cancer, anti-cancer of the stomach, leukemia, anti-neural cancer, inhibitor against colon carcinoma cells, anti-breast cancer, anti-cervical cancer and anti-carcinoma of the pancreas.
Described antitumor drug is the medicine of inhibition tumor cell proliferation activity.
Described antitumor drug is the medicine suppressing lung cell A549 and liver cancer cell SMCC-7721 proliferation activity.
Relative to prior art, beneficial effect of the present invention is:
The di-aryl urea compounds that fragrant heterocycleamide provided by the invention replaces, be a kind of novel compound with anti-tumor activity obtained through chemosynthesis, it has good inhibit activities to VEGFR-2 kinases, and can the proliferation activity of inhibition tumor cell.Scientific research has found that the generation of the generation of blood vessel and tumour, development and migration have substantial connection, suppress the formation of new vessel can the effectively growth of Tumor suppression and migration, many somatomedins all participate in the generation of new vessel, and wherein VEGFR-2 is the strongest known positive regulatory factor.The di-aryl urea compounds that fragrant heterocycleamide provided by the invention replaces is by suppressing the kinase whose activity of VEGFR-2, block the signal path of its induction, thus can the hyperplasia of inhibition tumor cell and migration, therefore the di-aryl urea compounds that fragrant heterocycleamide provided by the invention replaces can be used for antitumor drug and suppresses the preparation of VEGFR-2 kinase activity medicine, has a good application prospect and scientific research value.
The preparation method of the di-aryl urea compounds that fragrant heterocycleamide provided by the invention replaces, target compound can be obtained by the conventional organic synthesis of three steps, there is the advantages such as reaction process is simple to operate, raw material is easy to get, reaction conditions gentle, agents useful for same is cheap, be suitable for the scale operation manufacture of pharmacy corporation.
In addition, the di-aryl urea compounds that fragrant heterocycleamide provided by the invention replaces, to its cell-proliferation activity of inhibiting tumour cells comprising Non-small cell lung carcinoma cell (A549) and liver cancer cell (SMCC-7721), the preparation of antitumor drug can be can be applicable to.
Accompanying drawing explanation
Fig. 1 is the synthetic route chart of the di-aryl urea compounds that fragrant heterocycleamide provided by the invention replaces;
Wherein compound 1 is the bromo-PA of 5-, compound 2 is N-(5-bromopyridine-2-base)-2, 2-dimethylpropionamide, compound 3 is 5-bromine-2-methylpyridine, compound 4 is 5-bromopyridine-2-carboxylic acid, compound is 5 is 5-bromopyridine-2-carbonyl chlorine, compound 6 is the bromo-pyridine carboxamide of N-methyl-5-, compound 7 is 4-bromine-2-methylpyridine, compound 8 is 4-bromopyridine-2-carboxylic acid, compound 9 is 4-bromopyridine-2-carbonyl chlorine, compound 10 is the bromo-pyridine carboxamide of N-methyl-4-, compound 11 is p-aminophenyl borate hydrochlorate, compound 12 is N-[5-(4-aminophenyl) pyridine-2-base]-2, 2-dimethylpropionamide, compound 13 is 5-(4-aminophenyl)-N-picoline-2-methane amide, compound 14 is 4-(4-aminophenyl)-N-picoline-2-methane amide, compound 15 is containing disubstituted aniline, compound 16, 17 and 18 di-aryl urea compounds replaced for the fragrant heterocycleamide with anti-tumor activity.
What mark in figure is specially: a:(CH
3)
3cOCl, Et
3n, DCM, rt; B:KMnO
4, HCl; C:SOCl
2; D:CH
3nH
2, THF; E:dioxane, Pd (pddf) Cl
2, K
2cO
3, 100 DEG C; F:BTC, Et
3n, DCM.
Embodiment
The invention provides the di-aryl urea compounds that a kind of fragrant heterocycleamide with anti-tumor activity replaces, the di-aryl urea compounds that this fragrant heterocycleamide replaces has anti-tumor activity, can be applied to the preparation of antitumor drug.
Be described in detail the present invention below in conjunction with drawings and Examples, the explanation of the invention is not limited.
The di-aryl urea compounds that the fragrant heterocycleamide with anti-tumor activity provided by the invention replaces, its chemical structural formula is:
Wherein, Ar is the pyridine heterocycle containing pivaloyl amido or methylaminocarbonyl, R
1for trifluoromethyl, trifluoromethoxy or halogen, R
2the carbonatoms replaced by tertiary amine groups for trifluoromethyl, trifluoromethoxy, halogen or end is the alkoxyl group of 2 ~ 3, the carbonatoms that wherein end is replaced by tertiary amine groups be 2 ~ 3 alkoxyl group be connected to the contraposition of urea by O atom.Tertiary amine groups is dimethylamino, morpholinyl or piperidyl.
The preparation and method for screening active ingredients with the di-aryl urea compounds that antitumor drug candidate virtue heterocycleamide replaces is described in detail below in conjunction with the synthetic route shown in Fig. 1 and concrete synthesis example.
Embodiment 1
In the structural formula of this compound, Ar is the pyridine heterocycle containing pivaloyl amido, R
1for trifluoromethyl, R
2for bromine, by following steps preparation (see Fig. 1):
1) N-(5-bromopyridine-2-base)-2,2-dimethylpropionamide (compound 2) are prepared by 5-bromo-2-amido pyridine (compound 1)
Be dissolved in anhydrous methylene chloride by bromo-for 8.6g 5-2-amido pyridine (compound 1), add 25ml anhydrous triethylamine, ice bath stirs 30min, and then slowly drip the anhydrous methylene chloride solution of pivaloyl chloride 12ml, after dripping, room temperature reaction spends the night.With dichloromethane extraction reaction solution 2 ~ 3 times, then extraction liquid is washed with water 2 times, finally use saturated sodium bicarbonate solution, saturated common salt water washing successively, anhydrous sodium sulfate drying, be spin-dried for and obtain residue, be separated with chromatography column and obtain solid N-(5-bromopyridine-2-base)-2,2-dimethylpropionamide (compound 2) 7.3g, productive rate 90%;
2) by N-(5-bromopyridine-2-base)-2,2-dimethylpropionamide (compound 2) and p-aminophenyl borate hydrochlorate (compound 11) prepare N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide (compound 12) by Suzuki linked reaction
By N-(5-bromopyridine-2-base)-2,2-dimethylpropionamide (compound 2) 1.3g (5.3mmol), p-aminophenyl borate hydrochlorate (compound 11) 1g (5.8mmol), Anhydrous potassium carbonate 2.2g (15.8mmol), catalyst P d (pddf) Cl
20.6g (0.51mmol) is dissolved in Isosorbide-5-Nitrae-dioxane: in water=90mL:30mL, nitrogen protection, spends the night in 100 DEG C of reactions.Be cooled to room temperature, suction filtration, 1,4-dioxane washing leaching cake, collect filtrate, be spin-dried for obtain residue, be separated (eluting solvent is by sherwood oil: ethyl acetate=3:1) through chromatography column and obtain solid N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide (compound 12) 1.4g, productive rate about 75%;
Physico-chemical property: mp:159 ~ 161 DEG C, MS (EI) [M]+: m/z=269
Hydrogen spectrum nuclear magnetic resonance data:
1h NMR (400MHz, CDCl
3) δ 8.43 (d, J=2.2Hz, 1H), 8.34 (d, J=8.7Hz, 2H), 7.91 (m, 1H), 7.41 – 7.37 (m, 2H), 6.81 – 6.78 (m, 2H), 1.37 (s, 10H).
3) N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide (compound 12) and 5-bromo-3-5-trifluoromethylaniline (compound 15) prepare N-(5-(4-(3-(the bromo-5-of 3-(trifluoromethyl) phenyl) urea groups) phenyl) pyridine-2-base) pivalyl amine (compound 16) by condensation reaction
Under condition of ice bath, heavily steam methylene dichloride with 15mL two for 0.09g (0.3mmol) (trichloromethyl) carbonic ether (BTC) is dissolved and stir 5min, slowly drip the dichloromethane solution of the bromo-3-5-trifluoromethylaniline of 0.16g (0.7mmol) 5-(compound 15) again, dropwise and stir 15min, the dichloromethane solution 10mL dripping 0.12mL (0.89mmol) triethylamine is continued in turbid solution, dropwise rear continuation and stir 15min, then in reaction soln, drip N-[5-(4-aminophenyl) pyridine-2-base]-2, the dichloromethane solution 10mL of 2-dimethylpropionamide (compound 12) 0.2g (0.74mmol) and 0.12mL (0.89mmol) triethylamine, dropwise rear continuation and stir 20min, reaction solution is used saturated sodium bicarbonate solution successively, saturated common salt water washing, anhydrous sodium sulfate drying, be spin-dried for and obtain residue, be separated with chromatography column and obtain solid N-(5-(4-(3-(the bromo-5-of 3-(trifluoromethyl) phenyl) urea groups) phenyl) pyridine-2-base) pivalyl amine (compound 16) 0.16g, productive rate 37%,
Gained compound structure is as follows:
Physico-chemical property: mp:140 ~ 142 DEG C, MS (ESI) [M+H]+: m/z=535
Hydrogen spectrum nuclear magnetic resonance data is:
1h NMR (400MHz, CDCl
3) δ 8.42 (s, 1H), 8.15 (d, J=8.7Hz, 1H), 7.95 (s, 1H), 7.66 (s, 1H), 7.44 (d, J=6.6Hz, 1H), 7.37 (d, J=8.6Hz, 2H), 7.32 (d, J=8.7Hz, 2H), 1.43 (s, 9H).
Embodiment 2
In the structural formula of this compound, Ar is the pyridine heterocycle containing pivaloyl amido, R
1for trifluoromethoxy, R
2for bromine.
Step 1) ~ 2) with step 1 in embodiment 1) ~ 2) identical, namely N-(5-bromopyridine-2-base)-2 is prepared by 5-bromo-2-amido pyridine (compound 1), 2-dimethylpropionamide (compound 2), N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide (compound 12) are prepared by Suzuki linked reaction again with p-aminophenyl borate hydrochlorate (compound 11).
3) by N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide (compound 12) and 4-bromo-2-trifluoro-methoxyaniline (compound 15) prepare N-(5-(4-(3-(the bromo-2-of 4-(trifluoromethoxy) phenyl) urea groups) phenyl) pyridine-2-base) pivalyl amine (compound 16) by condensation reaction, and concrete operation steps is:
Under condition of ice bath, heavily steam methylene dichloride with 15mL two for 0.13g (0.44mmol) (trichloromethyl) carbonic ether (BTC) is dissolved and stir 5min, slowly drip the dichloromethane solution of the bromo-2-trifluoro-methoxyaniline of 0.28g (1.1mmol) 4-again, dropwise and stir 15min, the dichloromethane solution 10mL dripping 0.2mL (1.32mmol) triethylamine is continued in turbid solution, dropwise rear continuation and stir 15min, then in reaction soln, drip N-[5-(4-aminophenyl) pyridine-2-base]-2, the dichloromethane solution 10mL of 2-dimethylpropionamide 0.3g (1.1mmol) and 0.2mL (1.32mmol) triethylamine, dropwise rear continuation and stir 20min, reaction solution is used saturated sodium bicarbonate solution successively, saturated common salt water washing, anhydrous sodium sulfate drying, be spin-dried for and obtain residue, be separated with chromatography column and obtain solid N-(5-(4-(3-(the bromo-2-of 4-(trifluoromethoxy) phenyl) urea groups) phenyl) pyridine-2-base) pivalyl amine (compound 16) 0.17g, productive rate is about 35%,
Gained compound structure is as follows:
Physico-chemical property: mp:202 ~ 204 DEG C, MS (ESI) [M+H]+: m/z=553
Nuclear magnetic resonance data is:
1h NMR (400MHz, CDCl
3) δ 8.43 (s, 1H), 8.37 (d, J=8.7Hz, 1H), 8.25 (d, J=8.8Hz, 1H), 7.92 (d, J=6.8Hz, 1H), 7.50 (q, J=7.3Hz, 4H), 7.44 (m, 1H), 7.41 (s, 1H), 1.39 (s, 9H).
Embodiment 3
In the structural formula of this compound, Ar is the pyridine heterocycle containing pivaloyl amido, R
1, R
2for chlorine.
Step 1) ~ 2) with step 1 in embodiment 1) ~ 2) identical, namely N-(5-bromopyridine-2-base)-2 is prepared by 5-bromo-2-amido pyridine (compound 1), 2-dimethylpropionamide (compound 2), N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide (compound 12) are prepared by Suzuki linked reaction again with p-aminophenyl borate hydrochlorate (compound 11).
3) by N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide (compound 12) and 3,4-dichlorphenamide bulk powder (compound 15) prepares N-(5-(4-(3-(3 by condensation reaction, 4-dichlorophenyl) urea groups) phenyl) pyridine-2-base) pivalyl amine (compound 16), concrete operation steps is:
Under condition of ice bath, heavily steam methylene dichloride with 15mL two for 0.18g (0.6mmol) (trichloromethyl) carbonic ether (BTC) is dissolved and stir 5min, slowly drip 0.24g (1.5mmol) 3 again, the dichloromethane solution of 4-dichlorphenamide bulk powder, dropwise and stir 15min, the dichloromethane solution 10mL dripping 0.25mL (1.8mmol) triethylamine is continued in turbid solution, dropwise rear continuation and stir 15min, then in reaction soln, drip N-[5-(4-aminophenyl) pyridine-2-base]-2, the dichloromethane solution 10mL of 2-dimethylpropionamide 0.3g (1.1mmol) and 0.25mL (1.8mmol) triethylamine, dropwise rear continuation and stir 20min, reaction solution is used saturated sodium bicarbonate solution successively, saturated common salt water washing, anhydrous sodium sulfate drying, be spin-dried for and obtain residue, be separated with chromatography column and obtain solid N-(5-(4-(3-(3, 4-dichlorophenyl) urea groups) phenyl) pyridine-2-base) pivalyl amine (compound 16) 0.16g, productive rate is about 32%,
Gained compound structure is as follows:
Physico-chemical property: mp:220 ~ 223 DEG C, MS (ESI) [M+H]+: m/z=459
Nuclear magnetic resonance data is:
1h NMR (400MHz, DMSO) δ 9.87 (s, 1H), 9.04 (s, 1H), 8.96 (s, 1H), 8.63 (d, J=1.8Hz, 1H), 8.13 (d, J=8.7Hz, 1H), 8.08 (d, J=2.4Hz, 1H), 7.90 (d, J=2.5Hz, 1H), 7.67 (d, J=8.7Hz, 2H), 7.57 (d, J=8.7Hz, 2H), 7.54 (d, J=8.8Hz, 1H), 7.35 (m, 1H), 1.26 (s, 9H).
Embodiment 4
In the structural formula of this compound, Ar is the pyridine heterocycle containing pivaloyl amido, R
1, R
2for trifluoromethyl.
Step 1) ~ 2) with step 1 in embodiment 1) ~ 2) identical, namely N-(5-bromopyridine-2-base)-2 is prepared by 5-bromo-2-amido pyridine (compound 1), 2-dimethylpropionamide (compound 2), N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide (compound 12) are prepared by Suzuki linked reaction again with p-aminophenyl borate hydrochlorate (compound 11).
3) by N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide (compound 12) and 3,5-bis-5-trifluoromethylaniline (compound 15) prepares N-(5-(4-(3-(3 by condensation reaction, 5-bis-(trifluoromethyl) phenyl) urea groups) phenyl) pyridine-2-base) pivalyl amine (compound 16), concrete operation steps is:
Under condition of ice bath, heavily steam methylene dichloride with 15mL two for 0.18g (0.6mmol) (trichloromethyl) carbonic ether (BTC) is dissolved and stir 5min, slowly drip 0.34g (1.5mmol) 3 again, the dichloromethane solution of 5-bis-5-trifluoromethylaniline, dropwise and stir 15min, the dichloromethane solution 10mL dripping 0.25mL (1.8mmol) triethylamine is continued in turbid solution, dropwise rear continuation and stir 15min, then in reaction soln, drip N-[5-(4-aminophenyl) pyridine-2-base]-2, the dichloromethane solution 10mL of 2-dimethylpropionamide 0.3g (1.1mmol) and 0.25mL (1.8mmol) triethylamine, dropwise rear continuation and stir 20min, reaction solution is used saturated sodium bicarbonate solution successively, saturated common salt water washing, anhydrous sodium sulfate drying, be spin-dried for and obtain residue, be separated with chromatography column and obtain solid N-(5-(4-(3-(3, 5-bis-(trifluoromethyl) phenyl) urea groups) phenyl) pyridine-2-base) pivalyl amine (compound 16) 0.14g, productive rate is about 24%,
Gained compound structure is as follows:
Physico-chemical property: mp:210 ~ 213 DEG C, MS (ESI) [M+H]+: m/z=524
Nuclear magnetic resonance data is:
1h NMR (400MHz, CDCl
3) δ 8.37 (s, 1H), 8.23 (s, 2H), 8.03 (s, 2H), 7.69 (d, J=8.9Hz, 2H), 7.55 (s, 1H), 7.36 (s, 4H), 1.45 (s, 9H).
Embodiment 5
In the structural formula of this compound, Ar is the pyridine heterocycle containing pivaloyl amido, R
1for fluorine, R
2for the alkoxyl group containing morpholinyl.
Step 1) ~ 2) with step 1 in embodiment 1) ~ 2) identical, namely N-(5-bromopyridine-2-base)-2 is prepared by 5-bromo-2-amido pyridine (compound 1), 2-dimethylpropionamide (compound 2), N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide (compound 12) are prepared by Suzuki linked reaction again with p-aminophenyl borate hydrochlorate (compound 11).
3) by N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide (compound 12) and the fluoro-4-of 3-(3-(morpholine-4-base) propoxy-) aniline (compound 15) prepare target compound (compound 16) by condensation reaction, and specific embodiment is:
Under condition of ice bath, heavily steam methylene dichloride with 15mL two for 0.18g (0.6mmol) (trichloromethyl) carbonic ether (BTC) is dissolved and stir 5min, slowly drip the dichloromethane solution of the fluoro-4-of 0.4g (1.5mmol) 3-(3-(morpholine-4-base) propoxy-) aniline again, dropwise and stir 15min, the dichloromethane solution 10mL dripping 0.25mL (1.8mmol) triethylamine is continued in turbid solution, dropwise rear continuation and stir 15min, then in reaction soln, drip N-[5-(4-aminophenyl) pyridine-2-base]-2, the dichloromethane solution 10mL of 2-dimethylpropionamide 0.3g (1.1mmol) and 0.25mL (1.8mmol) triethylamine, dropwise rear continuation and stir 20min, reaction solution is used saturated sodium bicarbonate solution successively, saturated common salt water washing, anhydrous sodium sulfate drying, be spin-dried for and obtain residue, be separated with chromatography column and obtain solid 0.14g (compound 16), productive rate is about 25.5%,
Gained compound structure is as follows:
Physico-chemical property: mp:193 ~ 196 DEG C, MS (ESI) [M+H]+: m/z=550
Nuclear magnetic resonance data is:
1h NMR (400MHz, DMSO) δ 9.87 (s, 1H), 8.81 (s, 1H), 8.71 (s, 1H), 8.63 (d, J=1.9Hz, 1H), 8.12 (d, J=8.7Hz, 1H), 8.06 (m, 1H), 7.65 (d, J=8.7Hz, 2H), 7.56 (d, J=8.7Hz, 2H), 7.51 – 7.46 (m, 1H), 7.10 (s, 1H), 7.08 (s, 1H), 4.04 (t, J=6.4Hz, 2H), 3.60 – 3.56 (m, 4H), 2.42 (t, J=7.2Hz, 2H), 2.36 (s, 4H), 1.91 – 1.83 (m, 2H), 1.26 (s, 9H).
Embodiment 6
In the structural formula of this compound, Ar is the pyridine heterocycle containing methylaminocarbonyl, R
1for trifluoromethyl, R
2for chlorine, by following steps preparation (see Fig. 1):
1) 5-bromopyridine-2-carboxylic acid (compound 4) is prepared by 5-bromine-2-methylpyridine (compound 3) by oxidizing reaction.
In the 250ml three-necked flask that agitator, thermometer and prolong are housed, add water and the 6g 5-bromine-2-methylpyridine of 150ml, treat warming-in-water to 80 DEG C, add the KMnO of 24.8g in three batches
4, every minor tick 1h, isothermal reaction 3-4h after adding.After having reacted, suction filtration, obtains clear soln, with the pH to 4-5 of concentrated hydrochloric acid regulator solution, have a small amount of white solid to separate out, suction filtration gained filter cake is 5-bromopyridine-2-carboxylic acid (compound 4), after the aqueous solution is spin-dried for by decompression, add ethanol in proper amount, now also adularescent solid is separated out, and discards, again carry out suction filtration, filtrate be spin-dried for and namely can obtain whole products, finally obtain about 3g solid 5-bromopyridine-2-carboxylic acid (compound 4), productive rate is about 39%.
2) the bromo-pyridine carboxamide of N-methyl-5-(compound 6) is prepared by 5-bromopyridine-2-carboxylic acid (compound 4) by replacement and ammonification two-step reaction.
At N
2provide protection under, by 20mL SOCl
2be added drop-wise in 3g 5-bromopyridine-2-carboxylic acid (compound 4), after dropwising, treat that solution return is to clarification, reaction 2h.After reaction terminates, decompression is spin-dried for removing SOCl
2, the residue after being spin-dried for is dissolved in anhydrous tetrahydro furan, under the condition of ice bath, this anhydrous tetrahydrofuran solution is added drop-wise in 10mL methylamine solution, after dropwising, removes ice bath, make reaction react 2h at room temperature.After reaction terminates, water is added in above-mentioned solution, with ethyl acetate solution extraction (50mL × 3), with pressure reducing and steaming solution after anhydrous sodium sulfate drying, resistates chromatography column is separated, and (eluent is sherwood oil by volume: ethyl acetate=3:1) obtains the bromo-pyridine carboxamide of solid N-methyl-5-(compound 6) 1.67g, productive rate 60%;
3) 5-(4-aminophenyl)-N-picoline-2-methane amide (compound 13) is prepared by the bromo-pyridine carboxamide of N-methyl-5-(compound 6) and p-aminophenyl borate hydrochlorate (compound 11) by Suzuki linked reaction
The bromo-pyridine carboxamide of N-methyl-5-(compound 6) 1.67g (7.8mmol), p-aminophenyl borate hydrochlorate 1.5g (8.6mmol), Anhydrous potassium carbonate 3.2g (23.3mmol), catalyst P d (pddf) Cl 20.9g (0.78mmol) are dissolved in 1; 4-dioxane: in water=120mL:40mL; nitrogen protection, spends the night in 100 DEG C of reactions.Be cooled to room temperature, suction filtration, 1,4-dioxane washing leaching cake, collect filtrate, be spin-dried for obtain residue, be separated (eluting solvent is by sherwood oil: ethyl acetate=6:1) through chromatography column and obtain solid 5-(4-aminophenyl)-N-picoline-2-methane amide (compound 13) 1.73g, productive rate about 98%;
4) 5-(4-aminophenyl)-N-picoline-2-methane amide (compound 13) and 4-chloro-3-5-trifluoromethylaniline (compound 15) prepare N-methyl-5-(4-(3-(the chloro-3-of 4-(trifluoromethyl) phenyl) urea groups) phenyl) picolinamide (compound 17) by condensation reaction
Under condition of ice bath, heavily steam methylene dichloride with 15mL two for 0.5g (1.7mmol) (trichloromethyl) carbonic ether (BTC) is dissolved and stir 5min, slowly drip the dichloromethane solution of the chloro-3-5-trifluoromethylaniline of 0.84g (4.3mmol) 4-again, dropwise and stir 15min, the dichloromethane solution 10mL dripping 0.7mL (5.2mmol) triethylamine is continued in turbid solution, dropwise rear continuation and stir 15min, then in reaction soln, drip the dichloromethane solution 10mL of 5-(4-aminophenyl)-N-picoline-2-methane amide (compound 13) 0.84g (3.7mmol) and 0.7mL (5.2mmol) triethylamine, dropwise rear continuation and stir 20min, reaction solution is used saturated sodium bicarbonate solution successively, saturated common salt water washing, anhydrous sodium sulfate drying, be spin-dried for and obtain residue, be separated with chromatography column and obtain solid N-methyl-5-(4-(3-(the chloro-3-of 4-(trifluoromethyl) phenyl) urea groups) phenyl) picolinamide (compound 17) 0.36g, productive rate 19%,
Gained compound structure is as follows:
Physico-chemical property: mp:224 ~ 225 DEG C, MS (ESI) [M+H]+: m/z=449
Nuclear magnetic resonance data is:
1h NMR (400MHz, DMSO) δ 9.32 (s, 1H), 9.16 (s, 1H), 8.92 (d, J=2.1Hz, 1H), 8.81 – 8.75 (m, 1H), 8.23-8.25 (m, 1H), 8.13 (d, J=2.3Hz, 1H), 8.07 (d, J=8.1Hz, 1H), 7.78 (d, J=8.7Hz, 2H), 7.67 – 7.64 (m, 2H), 7.63 (d, J=2.9Hz, 1H), 2.85 (d, J=4.8Hz, 3H).
Embodiment 7
In the structural formula of this compound, Ar is the pyridine heterocycle of methylaminocarbonyl, R
1, R
2for fluorine.
Step 1) ~ 3) with step 1 in embodiment 6) ~ 3) identical, namely 5-bromopyridine-2-carboxylic acid (compound 4) is prepared by 5-bromine-2-methylpyridine (compound 3), then the bromo-pyridine carboxamide of N-methyl-5-(compound 6) is prepared by 5-bromopyridine-2-carboxylic acid (compound 4) by replacement and ammonification two-step reaction, the bromo-pyridine carboxamide of N-methyl-5-(compound 6) and p-aminophenyl borate hydrochlorate (compound 11) prepare 5-(4-aminophenyl)-N-picoline-2-methane amide (compound 13) by Suzuki linked reaction.
4) 5-(4-aminophenyl)-N-picoline-2-methane amide (compound 13) and 3,4-difluoroaniline (compound 15) prepares N-methyl-5-(4-(3-(3,4-difluorophenyl) urea groups) phenyl) picolinamide (compound 17) by condensation reaction
Under condition of ice bath, heavily steam methylene dichloride with 15mL two for 0.3g (1mmol) (trichloromethyl) carbonic ether (BTC) is dissolved and stir 5min, slowly drip 0.3g (2.5mmol) 3 again, the dichloromethane solution of 4-difluoroaniline, dropwise and stir 15min, the dichloromethane solution 10mL dripping 0.4mL (3mmol) triethylamine is continued in turbid solution, dropwise rear continuation and stir 15min, then in reaction soln, drip the dichloromethane solution 10mL of 5-(4-aminophenyl)-N-picoline-2-methane amide (compound 13) 0.3g (1.32mmol) and 0.4mL (3mmol) triethylamine, dropwise rear continuation and stir 20min, reaction solution is used saturated sodium bicarbonate solution successively, saturated common salt water washing, anhydrous sodium sulfate drying, be spin-dried for and obtain residue, be separated with chromatography column and obtain solid N-methyl-5-(4-(3-(3, 4-difluorophenyl) urea groups) phenyl) picolinamide (compound 17) 0.12g, productive rate 18%,
Gained compound structure is as follows:
Physico-chemical property: mp:240 ~ 244 DEG C, MS (ESI) [M+H]+: m/z=382
Nuclear magnetic resonance data is:
1h NMR (400MHz, DMSO) δ 8.99 (d, J=5.1Hz, 2H), 8.92 (d, J=2.2Hz, 1H), 8.79 (d, J=4.9Hz, 1H), 8.24 (m, 1H), 8.07 (d, J=8.2Hz, 1H), 7.80 – 7.76 (m, 2H), 7.72 – 7.66 (m, 1H), 7.63 (t, J=5.5Hz, 2H), 7.37 (d, J=10.5Hz, 1H), 7.18 – 7.13 (m, 1H), 2.85 (d, J=4.8Hz, 3H).
Embodiment 8
In the structural formula of this compound, Ar is the pyridine heterocycle of methylaminocarbonyl, R
1for trifluoromethyl, R
2for chlorine, by following steps preparation (see Fig. 1):
1) 4-bromopyridine-2-carboxylic acid (compound 8) is prepared by 4-bromine-2-methylpyridine (compound 7) by oxidizing reaction.
In the 250mL three-necked flask that agitator, thermometer and prolong are housed, add water and the 4g 4-bromine-2-methylpyridine of 100ml, treat warming-in-water to 80 DEG C, add the KMnO of 16.6g in three batches
4, every minor tick 1h, isothermal reaction 3-4h after adding.After having reacted, suction filtration, obtains clear soln, with the pH to 4-5 of concentrated hydrochloric acid regulator solution, have a small amount of white solid to separate out, suction filtration gained filter cake is 4-bromopyridine-2-carboxylic acid (compound 8), after the aqueous solution is spin-dried for by decompression, add ethanol in proper amount, now also adularescent solid is separated out, and discards, again carry out suction filtration, filtrate be spin-dried for and namely can obtain whole products, finally obtain about 1.7g solid 4-bromopyridine-2-carboxylic acid (compound 8), productive rate is about 33%.
2) the bromo-pyridine carboxamide of N-methyl-4-(compound 10) is prepared by 4-bromopyridine-2-carboxylic acid (compound 8) by replacement and ammonification two-step reaction.
At N
2provide protection under, by 15mL SOCl
2be added drop-wise in 1.7g 4-bromopyridine-2-carboxylic acid, after dropwising, treat that solution return is to clarification, reaction 2h.After reaction terminates, decompression is spin-dried for removing SOCl
2, the residue after being spin-dried for is dissolved in anhydrous tetrahydro furan, under the condition of ice bath, this anhydrous tetrahydrofuran solution is added drop-wise in 8mL methylamine solution, after dropwising, removes ice bath, make reaction react 2h at room temperature.After reaction terminates, water is added in above-mentioned solution, with ethyl acetate solution extraction (50mL × 3), with pressure reducing and steaming solution after anhydrous sodium sulfate drying, resistates chromatography column is separated, and (eluent is sherwood oil by volume: ethyl acetate=3:1) obtains the bromo-pyridine carboxamide of solid N-methyl-4-(compound 10) 0.87g, productive rate 52%;
3) 4-(4-aminophenyl)-N-picoline-2-methane amide (compound 14) is prepared by the bromo-pyridine carboxamide of N-methyl-4-(compound 10) and p-aminophenyl borate hydrochlorate (compound 11) by Suzuki linked reaction
The bromo-pyridine carboxamide of N-methyl-4-(compound 10) 0.87g (4mmol), p-aminophenyl borate hydrochlorate 0.84g (4.86mmol), Anhydrous potassium carbonate 1.7g (12mmol), catalyst P d (pddf) Cl 20.5g (0.41mmol) are dissolved in 1; 4-dioxane: in water=90mL:30mL; nitrogen protection, spends the night in 100 DEG C of reactions.Be cooled to room temperature, suction filtration, 1,4-dioxane washing leaching cake, collect filtrate, be spin-dried for obtain residue, be separated (eluting solvent is by sherwood oil: ethyl acetate=6:1) through chromatography column and obtain solid 4-(4-aminophenyl)-N-picoline-2-methane amide (compound 14) 0.96g, productive rate about 98%;
4) 4-(4-aminophenyl)-N-picoline-2-methane amide (compound 14) and 4-chloro-3-5-trifluoromethylaniline (compound 15) prepare N-methyl-4-(4-(3-(the chloro-3-of 4-(trifluoromethyl) phenyl) urea groups) phenyl) picolinamide (compound 18) by condensation reaction
Under condition of ice bath, heavily steam methylene dichloride with 15mL two for 0.4g (1.4mmol) (trichloromethyl) carbonic ether (BTC) is dissolved and stir 5min, slowly drip the dichloromethane solution of the chloro-3-5-trifluoromethylaniline of 0.68g (3.5mmol) 4-again, dropwise and stir 15min, the dichloromethane solution 10mL dripping 0.6mL (4.2mmol) triethylamine is continued in turbid solution, dropwise rear continuation and stir 15min, then in reaction soln, drip the dichloromethane solution 10mL of 4-(4-aminophenyl)-N-picoline-2-methane amide (compound 14) 0.66g (3mmol) and 0.6mL (4.2mmol) triethylamine, dropwise rear continuation and stir 20min, reaction solution is used saturated sodium bicarbonate solution successively, saturated common salt water washing, anhydrous sodium sulfate drying, be spin-dried for and obtain residue, be separated with chromatography column and obtain solid N-methyl-4-(4-(3-(the chloro-3-of 4-(trifluoromethyl) phenyl) urea groups) phenyl) picolinamide (compound 18) 0.07g, productive rate 5%,
Gained compound structure is as follows:
Physico-chemical property: mp:275 ~ 279 DEG C, MS (ESI) [M+H]+: m/z=449
Nuclear magnetic resonance data is:
1h NMR (400MHz, DMSO) δ 9.26 (s, 1H), 9.15 (s, 1H), 8.85 – 8.81 (m, 1H), 8.65 (d, J=5.1Hz, 1H), 8.27 (d, J=1.4Hz, 1H), 8.14 (d, J=2.3Hz, 1H), 7.90 (m, 1H), 7.85 (d, J=8.7Hz, 2H), 7.67 (s, 1H), 7.65 (s, 2H), 2.85 (d, J=4.8Hz, 3H).
Embodiment 9
In the structural formula of this compound, Ar is the pyridine heterocycle of methylaminocarbonyl, R
1, R
2for trifluoromethyl.
Step 1) ~ 3) with step 1 in embodiment 8) ~ 3) identical, namely 4-bromopyridine-2-carboxylic acid (compound 8) is prepared by 4-bromine-2-methylpyridine (compound 7), then the bromo-pyridine carboxamide of N-methyl-4-(compound 10) is prepared by 4-bromopyridine-2-carboxylic acid (compound 8) by replacement and ammonification two-step reaction, the bromo-pyridine carboxamide of N-methyl-4-(compound 10) and p-aminophenyl borate hydrochlorate (compound 11) prepare 4-(4-aminophenyl)-N-picoline-2-methane amide (compound 14) by Suzuki linked reaction.
4) 4-(4-aminophenyl)-N-picoline-2-methane amide (compound 14) and 3,5-bis-5-trifluoromethylaniline (compound 15) prepares N-methyl-4-(4-(3-(two (trifluoromethyl) phenyl of 3,5-) urea groups) phenyl) picolinamide (compound 18) by condensation reaction
Under condition of ice bath, heavily steam methylene dichloride with 15mL two for 0.3g (1mmol) (trichloromethyl) carbonic ether (BTC) is dissolved and stir 5min, slowly drip 0.3g (1.3mmol) 3 again, the dichloromethane solution of 5-bis-5-trifluoromethylaniline, dropwise and stir 15min, the dichloromethane solution 10mL dripping 0.4mL (3mmol) triethylamine is continued in turbid solution, dropwise rear continuation and stir 15min, then in reaction soln, drip the dichloromethane solution 10mL of 4-(4-aminophenyl)-N-picoline-2-methane amide (compound 14) 0.3g (1.32mmol) and 0.4mL (3mmol) triethylamine, dropwise rear continuation and stir 20min, reaction solution is used saturated sodium bicarbonate solution successively, saturated common salt water washing, anhydrous sodium sulfate drying, be spin-dried for and obtain residue, be separated with chromatography column and obtain solid N-methyl-4-(4-(3-(3, two (trifluoromethyl) phenyl of 5-) urea groups) phenyl) picolinamide (compound 18) 0.1g, productive rate 16%,
Gained compound structure is as follows:
Physico-chemical property: mp:270 ~ 272 DEG C, MS (ESI) [M+H]+: m/z=482
Nuclear magnetic resonance data is:
1h NMR (400MHz, DMSO) δ 9.50 (s, 1H), 9.31 (s, 1H), 8.84 (d, J=4.9Hz, 1H), 8.66 (d, J=5.2Hz, 1H), 8.28 (d, J=1.4Hz, 1H), 8.17 (s, 2H), 7.91 (m, 1H), 7.86 (d, J=8.8Hz, 2H), 7.70 – 7.67 (m, 3H), 2.86 (d, J=4.9Hz, 3H).
The generation of blood vessel and the generation of tumour, development and migration have substantial connection, suppress the formation of new vessel can the growth of effective Tumor suppression.Many somatomedins all participate in the generation of new vessel, and wherein VEGFR-2 is the strongest known positive regulatory factor.By suppressing VEGFR-2 kinase whose activity, block the signal path of its induction, can the hyperplasia of inhibition tumor cell and migration, and then reach the object for the treatment of tumour.
Anti-tumor activity experiment is carried out to the di-aryl urea compounds that fragrant heterocycleamide provided by the invention replaces below.
1, the di-aryl urea compounds that fragrant heterocycleamide replaces screens the kinase whose inhibit activities of VEGFR-2
Kinases VEGFR-2 and substrate A bltide is purchased from Signal-Chem company, and select the ADP-GlobTM Kinase Assays detection kit of Promega company to detect the Inhibiting enzyme activity of target compound, working method illustrates according to test kit carries out.
By ATP (10mM) buffer (2 ×) (Tris 80mM, MgCl
220mM, BSA 0.2mg/mL, DTT2mM) dilute buffer (2 ×) solution that 40 times are mixed with ATP (250 μMs); The mixing solutions ATP solution of 250 μMs and Abltide liquor capacity 1:1 being hybridly prepared into ATP (125 μMs)-Abltide (0.5 μ g/ μ l) is for subsequent use; VEGFR-2 kinase solution buffer (1 ×) (Tris 40mM, MgCl
210mM, BSA 0.1mg/mL, DTT 1mM) dilute buffer (1 ×) solution for standby that 66 times are mixed with VEGFR-2 (1.5ng/ μ l); Target compound and positive control drug (Sorafinib) buffer (1 ×) are mixed with 6 × 10 respectively
-5mol/L, 6 × 10
-6mol/L, 6 × 10
-7mol/L, 6 × 10
-8mol/L, 6 × 10
-9mol/L, 6 × 10
-10the sample solution of mol/L concentration gradient, on 384 orifice plates, every hole adds the mixing solutions of 2 μ L ATP-Abltide successively, 1 μ L sample solution, 2 μ L enzyme solution; Blank well adds the mixing solutions of 3 μ L damping fluids and 2 μ L ATP-Abltide; Control wells adds the mixing solutions of 2 μ L ATP-Abltide, 1 μ L damping fluid, and 2 μ L enzyme solution, finish, hatch 60min at 30 DEG C; Add ADP-Glo reagent 5 μ L, at 25 DEG C, hatch 40min; Add Kinase detection reagent 10 μ L, at 25 DEG C, hatch 30min.Adopt the chemoluminescence module of the multi-functional microplate reader of PerkinElmer to measure the luminous value in every hole, calculate target compound to the inhibiting rate of VEGFR-2 and IC
50.
The di-aryl urea compounds that fragrant heterocycleamide provided by the invention replaces is specifically as shown in table 1 to the result of VEGFR-2 kinase inhibitory activity:
The di-aryl urea compounds that the fragrant heterocycleamide of table 1 replaces is to the kinase whose IC50 of VEGFR-2
Can find out that the di-aryl urea compounds that fragrant heterocycleamide provided by the invention replaces has the good kinase whose activity of suppression VEGFR-2 by table 1, part of compounds is suitable with positive control drug BAY 43-9006 to the kinase whose suppression of VEGFR-2, even has some activity to be higher than positive control.Illustrate that the di-aryl urea compounds that fragrant heterocycleamide provided by the invention replaces can suppress to apply in the medicine of VEGFR-2 kinase activity in preparation.
2, the antitumor activity screening of the di-aryl urea compounds of fragrant heterocycleamide replacement
The di-aryl urea compounds that employing mtt assay checks fragrant heterocycleamide to replace is to the growth inhibitory activity of tumour cell:
The di-aryl urea compounds that fragrant heterocycleamide provided by the invention replaces has antitumor action, vitro inhibition proliferation activity is had to tumour cell, at lung carcinoma cell (A549), there is in liver cancer cell (SMCC-7721) clone the proliferation activity of inhibition tumor cell, may be used for the treatment to these cancers; Compared with positive drug BAY 43-9006 (Sorafinib), individual compound shows higher inhibition tumor cell proliferation activity.
To be in the lung carcinoma cell (A549) of logarithmic phase, liver cancer cell (SMCC-7721), the trysinization with 0.25% 3 ~ 5 minutes, being diluted to concentration after piping and druming is evenly 1 × 10
4~ 2 × 10
4the single cell suspension of individual/mL, is parallelly inoculated in 96 well culture plates, and every hole inoculation volume is 180 μ L; In 37 DEG C, 5%CO
2cultivate 24 hours in incubator; With the aqueous solution containing 0.25%DMSO (N, N-dimethyl sulfoxide (DMSO)) for negative control, take BAY 43-9006 as positive control, the di-aryl urea compounds (1 × 10 that the fragrant heterocycleamide that testing sample adds 4 different concns replaces
-4mol/L; 2 × 10
-5mol/L; 4 × 10
-6mol/L; 8 × 10
-7mol/L), each concentration establishes 3 multiple holes, continues cultivation 48 hours; Then every hole adds the MTT working fluid 22 μ L of 5mg/mL, mixing, 37 DEG C of incubators are hatched after 4 hours and are taken out, nutrient solution is abandoned in careful suction, every hole adds 150 μ L DMSO, vibration 10min, and euzymelinked immunosorbent assay (ELISA) detector measures 490nm place, each hole ultraviolet absorption value (OD value), then calculate cell inhibitory rate, and obtain IC50 value according to inhibiting rate; The calculation formula of cell inhibitory rate is:
Inhibiting rate %=(positive controls OD value-medication group OD value)/negative control cell OD value × 100%
Detected result shows: compared with negative control group, and the di-aryl urea compounds that fragrant heterocycleamide of the present invention replaces has In-vitro Inhibitory Effect in various degree to above-mentioned 2 kinds of tumour cells, and concrete outcome is as shown in table 2.
The di-aryl urea compounds that the fragrant heterocycleamide of table 2. replaces is to the IC50 (μM) of different cell strain
The suppression of result display section compound on tumor cell is suitable with positive control drug BAY 43-9006, even has some activity to be higher than positive control.Illustrate that the proliferation inhibiting effect of di-aryl urea compounds to tumour cell (lung cell A549 and liver cancer cell SMCC-7721) that fragrant heterocycleamide provided by the invention replaces is obvious, can apply preparing in antitumor drug.
And the structure of the di-aryl urea compounds that the higher fragrant heterocycleamide of above-mentioned activity replaces is specifically as shown in table 3:
The structural formula of the di-aryl urea compounds that the fragrant heterocycleamide that table 3 activity is higher replaces
Because VEGFR-2 kinases is to the growth of tumour cell, and the di-aryl urea compounds that fragrant heterocycleamide provided by the invention replaces is to the kinase whose restraining effect of VEGFR-2, therefore the di-aryl urea compounds that the fragrant heterocycleamide with anti-tumor activity provided by the invention replaces can be applied in the antitumor drug being target spot with VEGFR-2 kinases, not only comprise anti-lung cancer and the medicines resistant to liver cancer of above-mentioned citing, in like manner, the medicines such as anti-cancer of the stomach, leukemia, anti-neural cancer, inhibitor against colon carcinoma cells, anti-breast cancer, anti-cervical cancer and anti-carcinoma of the pancreas can also be comprised.
Claims (10)
1. a di-aryl urea compounds for fragrant heterocycleamide replacement, it is characterized in that, its structural formula is as follows:
Wherein, Ar is the pyridine heterocycle containing pivaloyl amido or methylaminocarbonyl, R
1for trifluoromethyl, trifluoromethoxy or halogen, R
2the carbonatoms replaced by tertiary amine groups for trifluoromethyl, trifluoromethoxy, halogen or end is the alkoxyl group of 2 ~ 3, the carbonatoms that wherein end is replaced by tertiary amine groups be 2 ~ 3 alkoxyl group be connected to the contraposition of urea by O atom.
2. the di-aryl urea compounds of fragrant heterocycleamide replacement according to claim 1, is characterized in that: what be connected with atom N in described tertiary amine groups is straight chained alkyl, cycloalkyl or epoxy alkyl.
3. the di-aryl urea compounds of fragrant heterocycleamide replacement according to claim 1, is characterized in that: described tertiary amine groups is dimethylamino, morpholinyl or piperidyl.
4. a preparation method for the di-aryl urea compounds of fragrant heterocycleamide replacement, is characterized in that, comprise the following steps:
1) 5-bromo-2-amido pyridine obtains N-(5-bromopyridine-2-base)-2,2-dimethylpropionamide by substitution reaction;
2) N-(5-bromopyridine-2-base)-2,2-dimethylpropionamide and p-aminophenyl borate hydrochlorate obtain N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide by Suzuki linked reaction;
3) N-[5-(4-aminophenyl) pyridine-2-base]-2,2-dimethylpropionamide obtain by condensation reaction the di-aryl urea compounds that fragrant heterocycleamide replaces with containing disubstituted aniline.
5. a preparation method for the di-aryl urea compounds of fragrant heterocycleamide replacement, is characterized in that, comprise the following steps:
1) 5-bromine-2-methylpyridine obtains 5-bromopyridine-2-carboxylic acid through oxidizing reaction, and then 5-bromopyridine-2-carboxylic acid is successively through substitution reaction and aminating reaction, the obtained bromo-pyridine carboxamide of N-methyl-5-;
Or 4-bromine-2-methylpyridine obtains 4-bromopyridine-2-carboxylic acid through oxidizing reaction, and then 4-bromopyridine-2-carboxylic acid is successively through substitution reaction and aminating reaction, the obtained bromo-pyridine carboxamide of N-methyl-4-;
2) the bromo-pyridine carboxamide of N-methyl-5-and p-aminophenyl borate hydrochlorate obtain 5-(4-aminophenyl)-N-picoline-2-methane amide by Suzuki linked reaction;
Or the bromo-pyridine carboxamide of N-methyl-4-and p-aminophenyl borate hydrochlorate obtain 4-(4-aminophenyl)-N-picoline-2-methane amide by Suzuki linked reaction;
3) 5-(4-aminophenyl)-N-picoline-2-methane amide obtains by condensation reaction the di-aryl urea compounds that fragrant heterocycleamide replaces with containing disubstituted aniline;
Or 4-(4-aminophenyl)-N-picoline-2-methane amide obtains by condensation reaction the di-aryl urea compounds that fragrant heterocycleamide replaces with containing disubstituted aniline.
6. the di-aryl urea compounds that the fragrant heterocycleamide in claim 1-3 described in any one replaces suppresses the application in VEGFR-2 kinase activity medicine in preparation.
7. the di-aryl urea compounds that the fragrant heterocycleamide in claim 1-3 described in any one replaces is preparing the application in antitumor drug.
8. apply as claimed in claim 7, it is characterized in that: the medicine that described antitumor drug is is target spot with VEGFR-2 kinases, comprise the medicine of anti-lung cancer, anti-liver cancer, anti-cancer of the stomach, leukemia, anti-neural cancer, inhibitor against colon carcinoma cells, anti-breast cancer, anti-cervical cancer and anti-carcinoma of the pancreas.
9. apply as claimed in claim 7, it is characterized in that: described antitumor drug is the medicine of inhibition tumor cell proliferation activity.
10. apply as claimed in claim 9, it is characterized in that: described antitumor drug is the medicine suppressing lung cell A549 and liver cancer cell SMCC-7721 proliferation activity.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105924387A (en) * | 2016-04-28 | 2016-09-07 | 西安交通大学 | Diarylurea compound with antitumor activity, and preparation method and application thereof |
| CN106236751A (en) * | 2016-07-28 | 2016-12-21 | 浙江工业大学 | The application in preparing cancer therapy drug of a kind of benzamide derivatives containing urea bridge |
| CN106748989A (en) * | 2016-11-14 | 2017-05-31 | 西安交通大学 | A kind of diaryl urea compound with antitumor activity and its preparation method and application |
| CN106748991A (en) * | 2016-11-14 | 2017-05-31 | 西安交通大学 | A kind of double aryl carbamide compounds with antitumor activity and its preparation method and application |
| CN106748990A (en) * | 2016-11-14 | 2017-05-31 | 西安交通大学 | A kind of substituted bisarylurea compound with antitumor activity and its preparation method and application |
| CN109503553A (en) * | 2018-12-05 | 2019-03-22 | 西安交通大学 | A kind of light Affinity Probes molecule and preparation method thereof based on VEGFR-2 inhibitor B14 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999032436A1 (en) * | 1997-12-22 | 1999-07-01 | Bayer Corporation | Inhibition of raf kinase using symmetrical and unsymmetrical substituted diphenyl ureas |
| CN103288684A (en) * | 2013-05-03 | 2013-09-11 | 西安交通大学 | Biphenyl carbamide compound with antineoplastic activity and preparation method thereof |
| CN103936631A (en) * | 2014-04-14 | 2014-07-23 | 西安交通大学 | Oximido diphenyl urea compound as well as preparation method and application thereof |
-
2015
- 2015-03-11 CN CN201510107596.XA patent/CN104817493A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999032436A1 (en) * | 1997-12-22 | 1999-07-01 | Bayer Corporation | Inhibition of raf kinase using symmetrical and unsymmetrical substituted diphenyl ureas |
| CN103288684A (en) * | 2013-05-03 | 2013-09-11 | 西安交通大学 | Biphenyl carbamide compound with antineoplastic activity and preparation method thereof |
| CN103936631A (en) * | 2014-04-14 | 2014-07-23 | 西安交通大学 | Oximido diphenyl urea compound as well as preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 段晓男等: "4-氨基嘧啶取代的双芳基脲类化合物的合成及抗肿瘤细胞增殖活性", 《中国药物化学杂志》 * |
Cited By (9)
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| CN105924387A (en) * | 2016-04-28 | 2016-09-07 | 西安交通大学 | Diarylurea compound with antitumor activity, and preparation method and application thereof |
| CN106236751A (en) * | 2016-07-28 | 2016-12-21 | 浙江工业大学 | The application in preparing cancer therapy drug of a kind of benzamide derivatives containing urea bridge |
| CN106748989A (en) * | 2016-11-14 | 2017-05-31 | 西安交通大学 | A kind of diaryl urea compound with antitumor activity and its preparation method and application |
| CN106748991A (en) * | 2016-11-14 | 2017-05-31 | 西安交通大学 | A kind of double aryl carbamide compounds with antitumor activity and its preparation method and application |
| CN106748990A (en) * | 2016-11-14 | 2017-05-31 | 西安交通大学 | A kind of substituted bisarylurea compound with antitumor activity and its preparation method and application |
| CN106748991B (en) * | 2016-11-14 | 2020-03-17 | 西安交通大学 | Diaryl urea compound with anti-tumor activity and preparation method and application thereof |
| CN106748989B (en) * | 2016-11-14 | 2020-03-17 | 西安交通大学 | Diaryl urea compound with anti-tumor activity and preparation method and application thereof |
| CN109503553A (en) * | 2018-12-05 | 2019-03-22 | 西安交通大学 | A kind of light Affinity Probes molecule and preparation method thereof based on VEGFR-2 inhibitor B14 |
| CN109503553B (en) * | 2018-12-05 | 2020-08-28 | 西安交通大学 | Light affinity probe molecule based on VEGFR-2 inhibitor B14 and preparation method thereof |
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