CN104876878A - 5-aryl phenol-2 alkyl substituted urea benzimidazole compound and applications thereof - Google Patents

5-aryl phenol-2 alkyl substituted urea benzimidazole compound and applications thereof Download PDF

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CN104876878A
CN104876878A CN201410069823.XA CN201410069823A CN104876878A CN 104876878 A CN104876878 A CN 104876878A CN 201410069823 A CN201410069823 A CN 201410069823A CN 104876878 A CN104876878 A CN 104876878A
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benzimidazolyl
radicals
phenoxy group
phenyl
alkyl substituted
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CN104876878B (en
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丁克
程辉敏
李迎君
谈理
王雪聪
张章
庄晓曦
龙活尤
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Guangzhou Institute of Biomedicine and Health of CAS
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Guangzhou Institute of Biomedicine and Health of CAS
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D235/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings
    • C07D235/02—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings condensed with carbocyclic rings or ring systems
    • C07D235/04—Benzimidazoles; Hydrogenated benzimidazoles
    • C07D235/24—Benzimidazoles; Hydrogenated benzimidazoles 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 in position 2
    • C07D235/30—Nitrogen atoms not forming part of a nitro radical

Abstract

The invention discloses a kind of -2 alkyl substituted urea benzimidazoles compound of 5- aryl phenol and its application, the compound has formula (I) structure. - 2 alkyl substituted urea benzimidazoles compound of 5- aryl phenol of the present invention can significantly inhibit the proliferation of kinds of tumor cells in vitro, and the retardance for forming the micro-pipe of tumour cell with the cell cycle G2/M phase shows dose dependent; New medicament selection is provided for clinical treatment tumour simultaneously.

Description

5-aryl phenol-2 alkyl substituted urea benzimidazoles compound and application thereof
Technical field
The present invention relates to chemical medicine, particularly relate to a kind of 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound and application thereof.
Background technology
Tumour, especially malignant tumour are the major diseases of serious threat human health and life security.According to statistics, malignant tumour caused the whole world 8,200,000 people's death in 2012; And it is predicted, in following 20 years, the annual cases of cancer in the whole world is by by 1,400 ten thousand to rise to 2,200 ten thousand (Globocan2012, IARC) in 2012.In China, tumour is one of most important lethal cause of disease equally.Therefore, finding and find new effective antitumour medicine is the important problem of Present Global.
Microtubule inhibitors is important antitumor drug, which includes microtubule stabilizer and microtubule destabilizer.If microtubule inhibitors drug main is by plant origin, as vinealeucoblastine(VLB) analogue and paclitaxel analogs.These medicines all have complicated chemical structure, poor solubility, and resistance problems.The small molecules microtubule inhibitors of development of new, is expected to the deficiency solving existing medicine.
The people such as ErnestLacey find (BiochemicalPharmacology, 34,7,1073-1077; BiochemicalPharmacology, 34,19,3603-3605) benzimidazoles compound has the activity better suppressing tubulin and the growth suppressing associated tumor cells.
Summary of the invention
Based on this, be necessary to provide a kind of anti-tumor activity and good 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound of solubleness.
A kind of 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound, has formula I structure:
Wherein, Ar is naphthyl or phenyl, and described phenyl is optional from fluorine for appointing by 0,1,2 or 3, chlorine, bromine, iodine, methyl, ethyl, sec.-propyl, hydroxyl, methoxyl group, oxyethyl group, trifluoromethyl, cyano group, alkynyl, phenyl ring, and benzyloxy substituents replaces;
R is methyl or ethyl.
Wherein in some embodiments, when described Ar is phenyl, the 2nd, 6 of substituting group not substituted-phenyl
Wherein in some embodiments, described Ar is phenyl, and described phenyl is optional from fluorine for appointing by 0,1,2 or 3, chlorine, bromine, iodine, methyl, ethyl, sec.-propyl, hydroxyl, methoxyl group, oxyethyl group, trifluoromethyl, cyano group, alkynyl, phenyl ring, and benzyloxy substituents replaces;
Described R is methyl.
Wherein in some embodiments, described phenyl is selected from chlorine, sec.-propyl for appointing, and hydroxyl substituent replaces.
Wherein in some embodiments, described compound is selected from:
1-(5-phenoxy group-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-phenoxy group-1H-benzimidazolyl-2 radicals)-3-ethyl carbamide;
1-(5-(2-chlorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-chlorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(4-chlorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(2-methoxyphenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-methoxyphenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(4-methoxyphenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3,4-dimethoxy phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3,5-dimethoxy phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3,4,5-trimethoxy phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-fluorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(5-fluorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-bromine phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-methylphenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-4-trifluoromethylphenopendant)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-ethyl phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-sec.-propyl phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-cyano-benzene oxygen)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-phenyl biphenylyloxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(4-phenyl biphenylyloxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-ethynyl phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(naphthalene 1-oxygen base)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(4-hydroxyphenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(4-ethoxy phenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(4-benzyloxy phenoxyl)-1H-benzimidazolyl-2 radicals)-3-MU.
Another object of the present invention is to the application that a kind of 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound is provided.
A kind of 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound or its pharmacy acceptable salt are preparing the application in prevention and therapy tumour medicine.
Wherein in some embodiments, described tumour comprises liver cancer, leukemia, lung cancer, cervical cancer, colorectal carcinoma, cancer of the stomach, the esophageal carcinoma, ovarian cancer, mammary cancer, melanoma, sarcoma or carcinoma of the pancreas.
Another object of the present invention is to the composition that a kind of Prevention and tumor are provided.
A composition for Prevention and tumor, its activeconstituents is 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound described in claim 1-4 or its pharmacy acceptable salt.
Another object of the present invention is to the application that a kind of 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound is provided.
A kind of 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound or its pharmacy acceptable salt suppress the application in the retarding agent of the inhibitor of tubulin polymerization, mitotic inhibitor, chemotherapy, radiotherapeutic sensitizer, cell death inducing inductor, arresting cell cycle in preparation.
The present invention has following beneficial effect compared to prior art:
(1) 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound that the present invention relates to significantly can suppress the propagation of kinds of tumor cells in vitro, is formed and the retardance of cell cycle G2/M phase presents dose-dependently to the microtubule of tumour cell.
(2) 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound that the present invention relates to, for clinical treatment tumour provides new medicament selection, provide new selection for more effective, safer medicine further, contribute to solving the clinical treatment problems such as the resistance be on the rise.
(3) 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound that the present invention relates to, can be as medicine as follows.
(a), for the preparation of pharmaceutical composition, described pharmaceutical composition be used for inhibition tumor cell growth or treatment tumour;
(b), for the preparation of suppressing the inhibitor of tubulin polymerization;
(c), for the preparation of mitotic inhibitor;
(d), for the preparation of chemotherapy, radiotherapeutic sensitizer;
(e), for the preparation of cell death inducing inductor;
(f), retarding agent for the preparation of arresting cell cycle.
(4) 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound that the present invention relates to, raw material sources are wide, and cost is low.
Accompanying drawing explanation
Compound described in the embodiment 19 that Fig. 1 is different concns dosing 8 as a child afterwards on tumour cell NCI460 micro-tubular structure impact experimental result schematic diagram;
Compound described in the embodiment 19 that Fig. 2 is different concns in dosing 24 as a child afterwards to the experimental result schematic diagram of tumour cell NCI460 Cell cycle influences;
Compound described in the embodiment 1 that Fig. 3 is different concns in dosing 8 as a child afterwards to the experimental result schematic diagram of HepG2 cellular microtubules structure influence;
Compound described in the embodiment 1 that Fig. 4 is different concns in dosing 24 as a child afterwards to the experimental result schematic diagram of HepG2 Cell cycle influences.
Embodiment
The invention provides a kind of preparation method of 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound,
Comprise the steps, as shown in (II):
(1) be that raw material and the fluoro-2-N-methyl-p-nitroaniline of 5-react and generate 5-aryl-2-oil of mirbane amine compound under the effect of Anhydrous potassium carbonate with aryl phenol;
(2) step (1) products therefrom 5-aryl-2-oil of mirbane amine compound is in the nitroreduction condition 10% palladium carbon of routine, hydrogen (or iron powder, zinc powder reduction) reduction, generates 5-aryl-1,2-bis-amido benzene class compound;
(3) in step (2) gained reduzate 5-aryl-2-oil of mirbane amine compound, add 1.3-bis-acid methyl-2-sulfo-isourea, ring closure reaction generates 5-aryl benzimidazoles compound;
(4) step (3) products therefrom 5-aryl benzimidazoles compound reacts under microwave condition with alkylamine again, generates 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound.
Specifically will by following examples further instruction
Embodiment 1
1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
The synthesis of step 12-nitro-5-phenoxybenzamine
The phenol of 0.9mL (10mmol) is dissolved in 10mLN, in dinethylformamide, adds 1.4g (10mmol) Anhydrous potassium carbonate, 100 DEG C are stirred 1h.Add the fluoro-2-N-methyl-p-nitroaniline of 0.8g (5mmol) 5-subsequently, 100 DEG C are reacted about 6h, TLC detecting and tracking reaction, find that raw material reacts completely.Be cooled to after room temperature in impouring frozen water, separate out precipitation, obtain 1.1g crude product 2-nitro-5-phenoxybenzamine with Büchner funnel suction filtration, yield is 98.5%, and thick product is directly used in next step reaction without being further purified.
The synthesis of step 24-phenoxy group-1,2-phenylenediamine
1.1g (4.9mmol) 2-nitro-5-phenoxybenzamine that previous step is synthesized is dissolved in 10mL ethanol, adds 0.03g (0.24mmol) palladium carbon, argon replaces 3 times, hydrogen exchange 3 times, normal-temperature reaction is about 3h, is spin-dried for ethanol, add 20mL water, repeatedly extract by ethyl acetate, merge organic phase, dry, be spin-dried for solvent, column chromatography obtains 0.74g product 4-phenoxy group-1,2-phenylenediamine, and yield is 75.4%.
The synthesis of step 3 methyl-5-benzyloxy-1H-benzimidazole-2-carbamates
By 0.74g (3.7mmol) the 4-phenoxy group-1 that previous step is synthesized, 2-phenylenediamine is dissolved in 10mL ethanol, adds 1.2g (5.6mmol) 1,3-bis-carboxymethyl-2-methyl-2-sulfo-isourea, be heated to 95 DEG C of back flow reaction spend the night, reaction produces precipitation.Be cooled to room temperature, obtain 0.68g product methyl-5-benzyloxy-1H-benzimidazole-2-carbamates with Büchner funnel suction filtration, yield is 65.0%.1HNMR（400MHz，DMSO-d6）δ7.40(d,J=8.8Hz,1H),7.36-7.31(m,2H),7.08-7.04(m,2H),6.94-6.92(d,J=8.0Hz,2H),6.81-6.79(dd,J=8.8,2.0Hz,1H),3.74(s,3H)。
Step 41-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
Methyl-5-benzyloxy-1H-the benzimidazole-2-carbamates taking 0.6g (2.1mmol) adds in 5mL acetonitrile, 0.6mL (4.2mmol) 33% methylamine alcohol solution, 140 DEG C, microwave reaction 2h, in impouring frozen water, separate out precipitation, decompress filter obtains 0.37g product, and yield is 62.5%. 1HNMR（400MHz，DMSO-d6）δ11.51(s,1H),9.96(s,1H),7.36-7.31(m,2H),7.16(s,1H),7.06-7.02(m,2H),6.91(d,J=8.0Hz,2H),6.75(dd,J=8.4,1.6Hz,1H),2.74(d,J=4.4Hz,3H)。HRMS(ESI)，[M+H] +：283.1192。
Embodiment 2
1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-ethyl carbamide
Methyl-5-benzyloxy-1H-the benzimidazole-2-carbamates taking 0.6g (2.1mmol) adds in 5mL acetonitrile, 0.54mL (4.2mmol) 70% ethylamine solution, 140 DEG C, microwave reaction 2h, in impouring frozen water, separate out precipitation, decompress filter obtains 0.32g product 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals)-3-ethyl carbamide, yield is 51.0%. 1HNMR（400MHz，DMSO-d6）δ11.53(s,1H),9.85(s,1H),7.36-7.31(m,3H),7.20(s,1H),7.06-7.03(m,2H),6.91(d,J=8.4Hz,2H),6.75(d,J=8.4Hz,1H),3.22-3.18(m,2H),1.10(t,J=7.2Hz,3H)。HRMS(ESI)，[M+H] +：297.1347。
Embodiment 3
1-(5-(2-chlorophenoxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 2-chlorophenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 8.5%.Wherein the method for nitroreduction adopts iron powder reducing, and concrete operations are as follows: by 1.4g(24.6mmol) iron powder adds in 2mL water, instillation concentrated hydrochloric acid 0.5mL, 100 DEG C of backflow 1h.Add the 2-nitro-5-(2-chlorophenoxy be dissolved in 6mL ethanol) aniline 1.3g (4.9mmol), reaction is spent the night.Suction filtration while hot, filtrate adjusts pH to neutral with saturated solution of sodium bicarbonate, and have Precipitation, decompress filter, filtrate is spin-dried for solvent, and column chromatography obtains 1.0g product 1-(5-(2-chlorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU, yield is 87.2%. 1HNMR（400MHz，DMSO-d6）δ11.52(s,1H),9.97(s,1H),7.56(dd,J=8.0,2.0Hz,1H),7.36(d,J=8.0Hz,1H),7.30-7.26(m,1H),7.13-7.10(m,2H),6.97(s,1H),6.90(d,J=8.0Hz,2H),3.72(dd,J=8.4,2.0Hz,1H),2.74(d,J=4.4Hz,3H)。HRMS(ESI)，[M+H] +：317.0802。
Embodiment 4
1-(5-(3-chlorophenoxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 3-chlorophenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 30.0%.Wherein the method for nitroreduction adopts iron powder reducing, and concrete operations are as follows: by 1.2g(20.8mmol) iron powder adds in 2mL water, instillation concentrated hydrochloric acid 0.4mL, 100 DEG C of backflow 1h.Add the 2-nitro-5-(3-chlorophenoxy be dissolved in 6mL ethanol) aniline 1.1g(4.1mmol), reaction is spent the night.Suction filtration while hot, filtrate adjusts pH to neutral with saturated solution of sodium bicarbonate, and have Precipitation, decompress filter, filtrate is spin-dried for solvent, and column chromatography obtains 0.65g product 1-(5-(3-chlorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU, yield is 68.1%. 1HNMR（400MHz，DMSO-d6）δ7.38-7.32(m,2H),7.25(s,1H),7.10-7.06(m,2H),6.91-6.87(m,2H),6.77(dd,J=8.4,2.0Hz,1H),2.74(d,J=4.4Hz,3H)。HRMS(ESI)，[M+H] +：317.0803。
Embodiment 5
1-(5-(4-chlorophenoxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 4-chlorophenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 29.5%.Wherein the method for nitroreduction adopts iron powder reducing, and concrete operations are as follows: by 1.2g(21.2mmol) iron powder adds in 2mL water, instillation concentrated hydrochloric acid 0.5mL, 100 DEG C of backflow 1h.Add the 2-nitro-5-(4-chlorophenoxy be dissolved in 6mL ethanol) aniline 1.1g(4.1mmol), reaction is spent the night.Suction filtration while hot, filtrate adjusts pH to neutral with saturated solution of sodium bicarbonate, and have Precipitation, decompress filter, filtrate is spin-dried for solvent, and column chromatography obtains 0.6g product 1-(5-(4-chlorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU, yield is 62.7%. 1HNMR（400MHz，DMSO-d6）δ11.53(s,1H),9.97(s,1H),7.36(m,3H),7.13(s,1H),7.03(s,1H),6.93(d,J=8.8Hz,2H),6.76(dd,J=8.4,2.0Hz,1H),2.74(d,J=4.4Hz,3H)。HRMS(ESI)，[M+H] +：317.0801。
Embodiment 6
(1-(5-(2-methoxyphenoxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 2-methoxyphenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 14.4%. 1HNMR（400MHz，DMSO-d6）δ11.39(s,1H),9.90(s,1H),7.28(d,J=8.4Hz,1H),7.20-7.09(m,3H),6.93-6.87(m,3H),6.62(dd,J=8.4,2.4Hz,1H),3.76(s,3H),2.73(d,J=3.2Hz,3H)。HRMS(ESI)，[M+H] +：313.1296。
Embodiment 7
1-(5-(3-methoxyphenoxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 3-methoxyphenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 17.9%. 1HNMR（400MHz，DMSO-d6）δ11.51(s,1H),9.98(s,1H),7.35(d,J=8.4Hz,1H),7.21(t,J=4.0Hz,1H),7.15(s,1H),7.03(s,1H),6.76(dd,J=8.8,2.0Hz,1H),6.28(dd,J=8.4,2.0Hz,1H),6.49-6.44(m,2H),3.71(s,3H),2.74(d,J=4.8Hz,3H)。HRMS(ESI)，[M+H] +：313.1299。
Embodiment 8
1-(5-(4-methoxyphenoxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 4-methoxyphenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 20.5%. 1HNMR（400MHz，DMSO-d6）δ11.43(s,1H),9.91(s,1H),7.30(d,J=8.8Hz,1H),7.17(s,1H),6.91(s,5H),6.70(d,J=3.6Hz,1H),3.72(s,3H),2.74(d,J=4.4Hz,3H)。HRMS(ESI)，[M+H] +：313.1297。
Embodiment 9
1-(5-(3,4-dimethoxy phenoxy group)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 3,4-syringol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 21.7%. 1HNMR（400MHz，DMSO-d6）δ11.42(s,1H),9.91(s,1H),7.31(d,J=8.4Hz,1H),7.18(s,1H),6.95(s,1H),6.89(d,J=8.4Hz,2H),6.73-6.68(m,2H),6.40(dd,J=8.8,2.8Hz,1H),3.70(d,J=4.0Hz,6H),2.73(d,J=4.4Hz,3H)。HRMS(ESI)，[M+H] +：343.1403。
Embodiment 10
1-(5-(3,5-dimethoxy phenoxy group)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 3,5-syringol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 15.8%. 1HNMR（400MHz，DMSO-d6）δ11.52(s,1H),9.96(s,1H),7.34(d,J=8.0Hz,1H),7.15(s,1H),7.04(s,1H),6.75(d,J=8.0Hz,1H),6.20(s,1H),6.05(s,1H),3.68(s,6H),2.74(d,J=3.2Hz,3H)。HRMS(ESI)，[M+H] +：343.1402。
Embodiment 11
1-(5-(3,4,5-trimethoxy phenoxy group)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 3,4,5-trimethoxy phenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 31.5%. 1HNMR（400MHz，DMSO-d6）δ11.47(s,1H),9.94(s,1H),7.33(d,J=7.6Hz,1H),7.16(s,1H),7.00(s,1H),6.75(d,J=8.4Hz,1H),6.27(s,2H),3.67(s,6H),3.61(s,3H),2.73(d,J=4.0Hz,3H)。HRMS(ESI)，[M+H] +：373.1509。
Embodiment 12
1-(5-(3-fluorophenoxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 3-fluorophenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 17.1%.Wherein the method for nitroreduction adopts zinc powder reduction, and concrete operations are as follows: by 0.8g(3.2mmol) 2-nitro-5-(3-fluorophenoxy) aniline is dissolved in 10mL ethanol, adds 1.2g(16.0mmol) zinc powder; argon shield; slow instillation saturated ammonium chloride solution 8mL, normal-temperature reaction 6h
Decompress filter, filtrate is spin-dried for solvent, and column chromatography obtains product 1-(5-(3-fluorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU, yield is 93.2%. 1HNMR（400MHz，DMSO-d6）δ11.57(s,1H),9.98(s,1H),7.38-7.31(m,2H),7.12(s,1H),7.07(s,1H),6.87(t,J=8.0Hz,1H),6.78(d,J=8.4Hz,1H),6.73(d,J=8.0Hz,2H),2.74(d,J=3.2Hz,3H)。MS(ESI):m/z300.1[M+H] +。
Embodiment 13
1-(5-(5-fluorophenoxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 5-fluorophenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 20.5%.Wherein the method for nitroreduction adopts zinc powder reduction; concrete operations are as follows: by 1.0g(4.0mmol) 2-nitro-5-(4-fluorophenoxy) aniline is dissolved in 10mL ethanol; add 1.3g(20.0mmol) zinc powder; argon shield, slowly instillation saturated ammonium chloride solution 7mL, normal-temperature reaction 4h; decompress filter; filtrate is spin-dried for solvent, and column chromatography obtains product 1-(5-(5-fluorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU, yield is 95.2%. 1HNMR（400MHz，DMSO-d6）δ11.48(s,1H),9.95(s,1H),7.34(d,J=8.4Hz,1H),7.16(m,3H),6.97(m,3H),6.74(d,J=8.4Hz,1H),2.74(d,J=2.8Hz,3H)。MS(ESI):m/z300.1[M+H] +。
Embodiment 14
1-(5-(3-bromine phenoxy group)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 3-bromophenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 38.7%.Wherein the method for nitroreduction adopts zinc powder reduction; concrete operations are as follows: by 1.3g(4mmol) 2-nitro-5-(3-bromine phenoxy group) aniline is dissolved in 10mL methyl alcohol; add 1.3g(20mmol) zinc powder; argon shield, slowly instillation saturated ammonium chloride solution 8mL, normal-temperature reaction 2h; decompress filter; filtrate is spin-dried for solvent, and column chromatography obtains product 1-(5-(3-bromine phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU, yield is 71.7%.1HNMR（400MHz，DMSO-d6）δ7.65(s,1H),7.35(d,J=8.4Hz,1H),7.28(t,J=8.0Hz,1H),7.21(d,J=8.0Hz,1H),7.04(s,1H),6.93(dd,J=6.4,1.6Hz,1H),6.73(dd,J=6.4,2.0Hz,1H),2.75(d,J=4.4Hz,3H)。MS(ESI):m/z360.0[M+H] +。
Embodiment 15
1-(5-(3-methylphenoxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 3-methylphenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 24.4%.1HNMR（400MHz，DMSO-d6）δ11.50(s,1H),9.94(s,1H),7.34(d,J=8.4Hz,1H),7.22-7.16(m,2H),6.70(s,1H),6.86(d,J=7.6Hz,1H),6.75-6.70(m,3H),2.74(d,J=4.4Hz,3H),2.25(s,1H)。HRMS(ESI)，[M+H] +：
Embodiment 16
1-(5-(3-4-trifluoromethylphenopendant)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 3-trifloro methyl phenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 25.7%.1HNMR（400MHz，DMSO-d6）δ11.60(s,1H),9.99(s,1H),7.56(t,J=8.0Hz,1H),7.39(d,J=7.6Hz,1H),7.22-7.10(m,4H),6.81(d,J=8.4Hz,1H),2.74(d,J=3.2Hz,3H)。HRMS(ESI)，[M+H] +：351.1067。
Embodiment 17
1-(5-(3-ethyl phenoxy group)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 3-ethylphenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 7.8%.1HNMR（400MHz，DMSO-d6）δ11.49(s,1H),9.94s,1H),7.34(d,J=8.4Hz,1H),7.22(t,J=8.0Hz,1H),7.15(s,1H),7.00(s,1H),6.89(d,J=7.2Hz,1H),6.78(s,1H),6.74(d,J=8.4Hz,1H),6.70(d,J=8.4Hz,1H),2.74(d,J=2.4Hz,3H),2.55(q,J=7.2Hz,2H),1.14(t,J=7.2Hz,3H)。MS(ESI):m/z310.1[M+H] +
Embodiment 18
1-(5-(3-sec.-propyl phenoxy group)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 3-isopropyl-phenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 18.5%.1HNMR（400MHz，DMSO-d6）δ11.50(s,1H),9.94(s,1H),7.34(d,J=8.0Hz,1H),7.24-7.20(m,1H),7.16(s,1H),7.00(s,1H),6.93(d,J=7.6Hz,1H),6.74(d,J=8.4Hz,2H),6.67(d,J=8.0Hz,1H),2.87-2.80(m,1H),2.74(d,J=4.0Hz,3H),1.16(d,J=6.8Hz,6H)。HRMS(ESI)，[M+H] +：325.1660。
Embodiment 19
1-(5-(3-cyano-benzene oxygen)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 3-cyanophenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 24.8%.Wherein the method for nitroreduction adopts zinc powder reduction; concrete operations are as follows: by 1.0g(4.1mmol) 2-nitro-5-(3-cyano-benzene oxygen) aniline is dissolved in 10mL ethanol; add 1.4g(20.5mmol) zinc powder; argon shield, slowly instillation saturated ammonium chloride solution 6mL, normal-temperature reaction 4h; decompress filter; filtrate is spin-dried for solvent, and column chromatography obtains product 1-(5-(3-cyano-benzene oxygen)-1H-benzimidazolyl-2 radicals)-3-MU, yield is 90.9%.1HNMR（400MHz，DMSO-d6）δ11.60(s,1H),9.99(s,1H),7.55-7.51(m,2H),7.39-7.35(m,2H),7.24(d,J=6.8Hz,1H),7.18-7.03(m,2H),6.79(d,J=8.0Hz,1H),2.74(d,J=3.6Hz,3H)。HRMS(ESI)，[M+H] +：308.1144。
Embodiment 20
1-(5-(3-phenyl biphenylyloxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
The synthesis of step 13-phenyl xenol
By 0.54mL(5mmol) m-bromoacetophenone and 0.61g(5mmol) phenylo boric acid is dissolved in 5mL1; in 4-dioxane, add 0.5mL water, add 2.1g (15mmol) Anhydrous potassium carbonate and 0.2g (0.15mmol) tetra-triphenylphosphine palladium; argon shield, 70 DEG C of reaction 8h.Adding dilute hydrochloric acid adjusts pH to neutral, and repeatedly extract by ethyl acetate, merge organic phase, dry, be spin-dried for solvent, column chromatography obtains product 3-phenyl xenol 0.32g, productive rate 75.2%.
Step 21-(5-(3-phenyl biphenylyloxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 3-phenyl xenol for substrate, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 31.1%.1HNMR（400MHz，DMSO-d6）δ7.73(s,1H),7.44(d,J=7.2Hz,2H),7.41-7.32(m,6H),7.18(t,J=6.0Hz,1H),7.06(d,J=2.4Hz,1H),6.90(dd,J=8.0,2.0Hz,1H),6.76(dd,J=8.4,2.4Hz,1H),2.74(d,J=4.4Hz,3H)。HRMS(ESI)，[M+H] +：359.1500。
Embodiment 21
31-(5-(4-phenyl biphenylyloxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 4-phenyl xenol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 6.7%.1HNMR（400MHz，DMSO-d6）δ11.53(s,1H),9.96(s,1H),7.63-7.60(m,4H),7.43(t,J=7.6Hz,2H),7.37(d,J=8.0Hz,1H),7.32(t,J=7.2Hz,1H),7.15(s,1H),7.07(s,1H),6.99(d,J=8.8Hz,2H),6.80(dd,J=8.4,2.0Hz,1H),2.74(d,J=4.8Hz,3H)。MS(ESI):m/z359.2[M+H] +
Embodiment 22
1-(5-(3-ethynyl phenoxy group)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
The bromo-phenoxy group of step 12-nitro-5-(3-) synthesis of aniline
With 3-bromophenol for starting raw material, step is with the synthesis of 2-nitro-5--phenoxybenzamine, and yield is 67.3%.
Step 22-nitro-5-(3-trimethylsilyl acetylene base-phenoxy group) synthesis of aniline
The bromo-phenoxy group of 0.47g (1.5mmol) 2-nitro-5-(3-by previous step is synthesized) aniline and 0.23g (2.25mmol) trimethylsilyl acetylene be dissolved in 5mL acetonitrile; add 0.052g (0.075mmol) two triphenyl phosphorus palladium chloride, 0.014g (0.075mmol) cuprous iodide and 0.5mL (3mmol) N; N-diisopropylethylamine; argon shield, 70 DEG C of reaction 8h.Be cooled to room temperature, decompress filter, filtrate is spin-dried for, and adds 20mL water, repeatedly extracts by ethyl acetate, merges organic phase, and dry, be spin-dried for solvent, column chromatography obtains 0.4g product, and yield is 81.8%.
Step 32-nitro-5-(3-ethynyl-phenoxy group) synthesis of aniline
2-nitro-5-(3-trimethylsilyl acetylene base-phenoxy group by previous step is synthesized) aniline is dissolved in 10mL methyl alcohol, adds 0.34g (2.46mmol) Anhydrous potassium carbonate, room temperature reaction 2h.Be spin-dried for solvent, add 20mL water, repeatedly extract by ethyl acetate, merge organic phase, dry, be spin-dried for solvent, column chromatography obtains product 0.28g, and yield is 89.6%.
Step 44-(3-ethynyl-phenoxy group) synthesis of-1,2-phenylenediamine
2-nitro-5-(3-ethynyl-phenoxy group by previous step is synthesized) aniline is dissolved in 10mL methyl alcohol, adds 0.36g (5.5mmol) zinc powder, argon shield, slowly instillation 1.7mL saturated ammonium chloride solution, room temperature reaction 2h.Decompress filter, filtrate is spin-dried for, and uses 20mL acetic acid ethyl dissolution, elimination insolubles, and filtrate is spin-dried for, and column chromatography obtains product 0.23g, and yield is 93.3%.
Step 51-(5-(3-ethynyl phenoxy group)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 4-(3-ethynyl-phenoxy group)-1,2-phenylenediamine is for substrate, cyclization, amination steps are with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, two step total recoverys are 9.8%.1HNMR（400MHz，DMSO-d6）δ11.55(s,1H),9.98(s,1H),7.38-7.32(m,2H),7.15(d,J=7.6Hz,2H),7.05(s,1H),6.98(d,J=8.4Hz,1H),6.91(s,1H),6.77(d,J=7.2Hz,1H),4.18(s,1H),2.73(d,J=4.4Hz,3H)。MS(ESI):m/z306.1[M+H] +
Embodiment 23
1-(5-(naphthalene 1-oxygen base)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
Take naphthyl alcohol as starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 28.3%.1HNMR（400MHz，DMSO-d6）δ11.50(s,1H),9.95(s,1H),8.21(d,J=7.6Hz,1H),7.96(d,J=7.2Hz,1H),7.63(d,J=8.4Hz,1H),7.53-7.59(m,2H),7.42-7.37(m,2H),7.16(s,1H),7.04(s,1H),6.84(dd,J=8.4,2.0Hz,1H),6.79(d,J=7.6Hz,1H),2.73(d,J=4.4Hz,3H)。HRMS(ESI)，[M+H] +：333.1346。
Embodiment 24
1-(5-(4-hydroxyphenoxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
Take Resorcinol as starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 3.6%.1HNMR（400MHz，DMSO-d6）δ11.38(s,1H),9.90(s,1H),9.19(s,1H),7.28(d,J=8.4Hz,1H),7.19(s,1H),6.88(s,1H),6.81(d,J=8.8Hz,2H),6.73(d,J=8.8Hz,2H),6.67(dd,J=8.8,2.2Hz,2H),2.73(d,J=4.4Hz,3H)。MS(ESI):m/z298.1[M+H] +
Embodiment 25
1-(5-(4-ethoxy phenoxy)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 4-thanatol for starting raw material, step is with 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals) synthesis of-3-MU, total recovery is 30.6%.1HNMR（400MHz，DMSO-d6）δ11.42(s,1H),9.92(s,1H),7.30(d,J=8.8Hz,1H),7.18(s,1H),6.90(s,5H),6.70(dd,J=8.4,2.0Hz,1H),3.98(d,J=6.8Hz,2H),2.73(d,J=4.4Hz,3H),1.31(t,J=6.8Hz,3H)。MS(ESI):m/z326.1[M+H] +
Embodiment 26
1-(5-(4-benzyloxy phenoxyl)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU;
Step 12-nitro-5-(4-hydroxyphenoxy) synthesis of aniline
5.77g (52.5mmol) Resorcinol is dissolved in 30mLN, in dinethylformamide, add 7.24g (52.5mmol) Anhydrous potassium carbonate, 100 DEG C of reaction 30min, add the fluoro-2-N-methyl-p-nitroaniline of 2.8g (17.5mmol) 5-, reaction is spent the night.Be cooled to room temperature, in impouring frozen water, separate out solid, decompress filter, dry 2.1g product, yield is 48.8%.
Step 25-(4-benzyloxyphenoxy) synthesis of-2-N-methyl-p-nitroaniline
Get 0.1g (0.41mmol) 2-nitro-5-(4-hydroxyphenoxy of previous step synthesis) aniline is dissolved in 5mLN, in dinethylformamide, add 0.12g (0.82mmol) Anhydrous potassium carbonate and 0.11g (0.62mmol) bromobenzyl, room temperature reaction spends the night.Add 30mL water, repeatedly extract by ethyl acetate, merge organic phase, dry, be spin-dried for solvent and obtain 0.31g product, yield is 91.9%.
Step 31-(5-(4-benzyloxy phenoxyl)-1H-benzimidazolyl-2 radicals) synthesis of-3-MU
With 5-(4-benzyloxyphenoxy)-2-N-methyl-p-nitroaniline is substrate, nitroreduction, cyclization, amination steps 1-(5-(2-chlorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU, three step total recoverys are 26.6%. 1HNMR（400MHz，DMSO-d6）δ11.42(s,1H),9.91(s,1H),7.44-7.30(m,6H),7.18(br,1H),7.00(d,J=5.2Hz,2H),6.91(d,J=5.2Hz,2H),6.71(d,J=8.8Hz,1H),5.06(s,2H),2.74(d,J=4.4Hz,3H)。MS(ESI):m/z388.2[M+H] +
Embodiment 27
Present embodiments provide the compounds of this invention Proliferation of Tumor Cells In Vitro inhibit activities to test
Experimental technique
1) counting after digesting with different tumour cell Colo205 and NCI-H460 respectively, according to 1000-2000/hole, plants in 96 orifice plates, and each concentration establishes 6 multiple holes.
Go down to posterity after cell recovery and be no less than twice no more than 15 times.Observation of cell state before kind of plate, use cell state good, grow to about 6 seventy percent full, just to have changed liquid recently cell.
2) cultivate after 24 hours in cell culture incubator, 100 μ L compound working fluids are added, continue cultivation 72 hours.
Compound DMSO is made into the mother liquor of 10mol/l.10 μ L mother liquors are joined in 90 μ LDMSO, use turbula shaker mixing, stepwise dilution 5 times, is mixed with DMSO storage liquid successively, be positioned over 4 DEG C stand-by.Again mix before using and according to 1:500, storage liquid joined as working fluid in perfect medium, using turbula shaker mixing.Working fluid needs matching while using, causes separating out to prevent long-time placement.
3) suck substratum, add 100 μ L containing MTT(0.5mg/ml) substratum, place 4 hours in incubator.Substratum is removed clean, adds 100 μ LDMSO, concussion mixing.
The preparation of MTT: 1.25gMTT joins in 250mlPBS, and mixing, 4 DEG C keep in Dark Place.
4) porous plate microplate reader is used to detect the light absorption value of 570nm or 490nm.
5) the OD value prism software analysis detected also calculates the IC of each compound 50value.
Target compound determination of activity result is as shown in the table
Meanwhile, embodiment Isosorbide-5-Nitrae, 18, the IC50 of the compound on tumor cell K562 described in 19 is less than 50nM; 100nM is less than to the IC50 of tumour cell A431; 10 μMs are less than to the IC50 of tumour cell HepG2; 2 μMs are less than to the IC50 of tumour cell Hela; 100nM is less than to the IC50 of tumour cell MDA-MB-435S.Concrete data are as follows:
As can be seen from activity data above, the chemical combination in general formula of the present invention has the activity of very strong extracorporeal suppression tumor cell.And substituting group on phenyl ring and substituent electrical position etc. are little to the activity influence of compound, the compound that wherein the non-ortho position of phenyl ring replaces has higher anti-tumor activity.
Embodiment 28
The present embodiment is the compound 1-(5-(3-cyano-benzene oxygen described in the embodiment of the present invention 19)-1H-benzimidazolyl-2 radicals)-3-MU is to the experiment of tumour cell NCI460 micro-tubular structure and Cell cycle influences:
1, cell cycle analysis
(1) drug treating cell: the cell getting suitable concn is inoculated in 6 orifice plates, cell confluency degree is waited to add the cyano-benzene oxygen of compound 1-(5-(3-described in different concns embodiment 19 about 60% after 24h)-1H-benzimidazolyl-2 radicals)-3-MU medicine, continues to cultivate 24-48h.
(2) collecting cell: at predetermined time point collecting cell.Be transferred in 1.5EP pipe after cell being used trysinization, centrifugal 10 minutes of room temperature 1500rpm.More than twice is washed, centrifugal 10 minutes of 1500rpm with PBS.
First add the PBS of 300 μ L4 DEG C precoolings, after resuspended mixing, add the dehydrated alcohol 700 μ L of-20 DEG C of precoolings again, with the mixing of rifle head, put into-20 DEG C of refrigerators and fix more than 24 hours.
The centrifugal 10min of 3000rpm.PBS is used to wash 2 times, the centrifugal 10min of 3000rpm.Carry out resuspended (needing to ensure that cell concn is at about 10-6) with the PBS of about 200ul
Add RNase to the final concentration 50 μ g/ml of 50mg/ml, cell is placed in 37 DEG C of digestion 30min.
(3) staining analysis: add the PI dyestuff of preparation to final concentration 10 μ g/ml, room temperature lucifuge dyeing 30min.FCM analysis is carried out in 1h.Flow cytometer: FACSCalibur, BD.
Carry out data analysis.
2, immunofluorescence
(1) drug treating cell: the cell of suitable concn is inoculated in 6 orifice plates, waits cell confluency degree to add the medicine of different concns about 30%, continues to cultivate 6-24h after 24h.
(2) cell is fixed: in every hole, add the paraformaldehyde stationary liquid 300 μ L of 4%, ambient temperatare puts 30 minutes with fixed cell.
(3) cell-permeant: every hole adds 0.2%TritonX-100 penetrating liquid 300 μ L, and ambient temperatare puts 10 minutes to make cell membrane penetration.
(4) antigen blockade: normal goat serum (NormalGoatSerum, NGS) the 20 μ L of 3% is as confining liquid, and off-period, room temperature was spent the night for 1 hour or 4 DEG C, to remove the impact of intracellular non-specific binding.
(5) primary antibodie is hatched: the primary antibodie (a-tubulin) of immunofluorescence uses confining liquid (3%NGS) to dilute, Dilution ratio is determined (ordinary antibody generally dilutes with 1:50 endogenous antigen) according to antibody titer and born of the same parents' endoantigen abundance, incubated at room temperature 1.5 hours.
(6) two anti-hatch: the two anti-different choice goat antirabbit according to primary antibodie or goat anti-mouse antibodies, and select coupled FITC(green fluorescence according to actual requirement) or TRITC(red fluorescence); Use confining liquid (3%NGS) dilution, Dilution ratio is 1:100 ~ 1:200.Incubated at room temperature 1 hour.
(7) the dilution proportion DAPI stoste of core: 1 × PBS according to 1:4000 is contaminated, after sucking the washingbuffer in 24 orifice bores, the DAPI staining fluid of 200 μ L is added fast in every hole, after 5-6 second, absorb staining fluid after all adding at once and rapidly in every hole, add 500 μ L1 × PBS, clean twice according to step 3, note lucifuge.
(8) mounting: 80% glycerine dripping 2 μ L in darkroom on slide glass, tips upside down on cover glass on glycerine, slightly dries in the air and one to preserve after the meeting; Fluorescent microscope or laser confocal microscope detect.
As can be seen from Figure 1, do not adding in the compound situation described in embodiment 19, tumour cell NCI460 micro-tubular structure presents uniform sequential elongate, threadlike structure (shown in Figure 1A) outside nucleus, when the compound concentration described in embodiment 19 is from shown in 25nM(Figure 1B) be increased to shown in 75nM(Fig. 1 C), shown in 100nM(Fig. 1 D) time NCI460 micro-tubular structure occur that the filamentary texture of abnormal elongated ordered distribution disappears (shown in Figure 1B, C, D), cellular form does not occur simultaneously.This result proves that the compounds of this invention can affect the performance of the tubulin normal function of tumour cell in dose-dependent mode.
As can be seen from Figure 2, adding in the compound situation described in various dose embodiment 19, the cell that tumour cell NCI460 is in G0/G1 phase and S phase reduces gradually, and the cell being in the G2/M phase increases (shown in Fig. 2 A, B, C, D) gradually.This result proves that tumour cell can be arrested in the G2/M phase in dose-dependent mode by the compounds of this invention, thus the running affecting tumour cell normal cycle finally causes necrocytosis.
Embodiment 29
The present embodiment is the compound 1-(5-phenoxy group-1H-benzimidazolyl-2 radicals described in the embodiment of the present invention 1)-3-MU is on the experiment of the micro-tubular structure of HepG2 cell and the impact in cycle, and concrete experimental procedure is with reference to embodiment 28.
As can be seen from Figure 3, do not adding in the compound situation described in embodiment 1, HepG2 cellular microtubules structure presents uniform sequential elongate, threadlike structure (shown in Fig. 3 A) outside nucleus, when the compound concentration described in embodiment 1 is from shown in 0.04uM(Fig. 3 B) be increased to shown in 0.2uM(Fig. 3 C), shown in 1uM(Fig. 3 D) time HepG2 cellular microtubules structure occur that the filamentary texture of abnormal elongated ordered distribution disappears (shown in Fig. 3 B, C, D), cellular form does not occur simultaneously.This result proves that the compounds of this invention can affect the performance of the tubulin normal function of HepG2 cell in dose-dependent mode.
As can be seen from Figure 4, adding in the compound situation described in various dose embodiment 1, the cell that HepG2 cell is in G0/G1 phase and S phase reduces gradually, and the cell being in the G2/M phase increases (shown in Fig. 4 A, B, C, D) gradually.This result proves that tumour cell can be arrested in the G2/M phase in dose-dependent mode by the compounds of this invention, thus the running affecting tumour cell normal cycle finally causes necrocytosis.
In sum, 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound that the present invention relates to significantly can suppress the propagation of kinds of tumor cells in vitro, is formed and the retardance of cell cycle G2/M phase presents dose-dependently to the microtubule of tumour cell; Therefore the compound that the present invention relates to can be used for pharmaceutical compositions, described pharmaceutical composition is used for inhibition tumor cell growth or treatment tumour, for clinical treatment tumour provides new medicament selection, provide new selection for more effective, safer medicine further, contribute to solving the clinical treatment problems such as the resistance be on the rise.
The above embodiment only have expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but therefore can not be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.

Claims (9)

1. 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound, is characterized in that having formula I structure:
Wherein, Ar is naphthyl or phenyl, and described phenyl is optional from fluorine for appointing by 0,1,2 or 3, chlorine, bromine, iodine, methyl, ethyl, sec.-propyl, hydroxyl, methoxyl group, oxyethyl group, trifluoromethyl, cyano group, alkynyl, phenyl ring, and benzyloxy substituents replaces;
R is methyl or ethyl.
2. 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound according to claim 1, is characterized in that, when described Ar is phenyl, and the 2nd, 6 of substituting group not substituted-phenyl.
3. 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound according to claim 1 and 2, it is characterized in that, described Ar is phenyl, and described phenyl is for appointing by 0,1,2 or 3 optional from fluorine, chlorine, bromine, iodine, methyl, ethyl, sec.-propyl, hydroxyl, methoxyl group, oxyethyl group, trifluoromethyl, cyano group, alkynyl, phenyl ring, benzyloxy substituents replaces;
Described R is methyl.
4. 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound according to claim 1 and 2, is characterized in that, described phenyl is optional from chlorine, and sec.-propyl, hydroxyl substituent replaces.
5. 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound according to claim 1, it is characterized in that, described compound is selected from:
1-(5-phenoxy group-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-phenoxy group-1H-benzimidazolyl-2 radicals)-3-ethyl carbamide;
1-(5-(2-chlorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-chlorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(4-chlorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(2-methoxyphenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-methoxyphenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(4-methoxyphenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3,4-dimethoxy phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3,5-dimethoxy phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3,4,5-trimethoxy phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-fluorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(5-fluorophenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-bromine phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-methylphenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-4-trifluoromethylphenopendant)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-ethyl phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-sec.-propyl phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-cyano-benzene oxygen)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-phenyl biphenylyloxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(4-phenyl biphenylyloxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(3-ethynyl phenoxy group)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(naphthalene 1-oxygen base)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(4-hydroxyphenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(4-ethoxy phenoxy)-1H-benzimidazolyl-2 radicals)-3-MU;
1-(5-(4-benzyloxy phenoxyl)-1H-benzimidazolyl-2 radicals)-3-MU.
6. 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound described in Claims 1 to 5 any one or its pharmacy acceptable salt are preparing the application in prevention and therapy tumour medicine.
7. application according to claim 6, is characterized in that, described tumour comprises liver cancer, leukemia, lung cancer, cervical cancer, colorectal carcinoma, cancer of the stomach, the esophageal carcinoma, ovarian cancer, mammary cancer, melanoma, sarcoma or carcinoma of the pancreas.
8. a composition for Prevention and tumor, is characterized in that, its activeconstituents is 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound described in claim 1-4 or its pharmacy acceptable salt.
9. 5-aryl phenol-2 alkyl substituted urea benzimidazoles compound described in Claims 1 to 5 any one or its pharmacy acceptable salt suppress the application in the retarding agent of the inhibitor of tubulin polymerization, mitotic inhibitor, chemotherapy, radiotherapeutic sensitizer, cell death inducing inductor, arresting cell cycle in preparation.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20220051569A (en) * 2020-10-19 2022-04-26 고려대학교 산학협력단 Benzimidazole derivatives or its pharmaceutically acceptable salts and use thereof
CN115594667A (en) * 2022-10-18 2023-01-13 北京京佑奇康科技有限公司（Cn） Benzimidazole compound and preparation method and application thereof
CN115594667B (en) * 2022-10-18 2024-05-10 北京京佑奇康科技有限公司 Benzimidazole compound, and preparation method and application thereof

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1481368A (en) * 2000-12-15 2004-03-10 ��̩��˹ҩ��ɷ����޹�˾ Gyrase imhibitors and uses thereof
CN101538263A (en) * 2002-06-13 2009-09-23 沃泰克斯药物股份有限公司 2-ureido-6-heteroaryl-3H-benzoimidazole-4-carboxylic acid derivatives and related compounds as gyrase and/or topoisomerase IV inhibitors for the treatment of bacterial infections
CN101827827A (en) * 2007-10-19 2010-09-08 赛诺菲-安万特 6-aryl/heteroalkyloxy benzothiazole and benzimidazole derivatives, method for preparing same, application thereof as drugs, pharmaceutical compositions and novel use in particular as C-MET inhibitors

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1481368A (en) * 2000-12-15 2004-03-10 ��̩��˹ҩ��ɷ����޹�˾ Gyrase imhibitors and uses thereof
CN101538263A (en) * 2002-06-13 2009-09-23 沃泰克斯药物股份有限公司 2-ureido-6-heteroaryl-3H-benzoimidazole-4-carboxylic acid derivatives and related compounds as gyrase and/or topoisomerase IV inhibitors for the treatment of bacterial infections
CN101827827A (en) * 2007-10-19 2010-09-08 赛诺菲-安万特 6-aryl/heteroalkyloxy benzothiazole and benzimidazole derivatives, method for preparing same, application thereof as drugs, pharmaceutical compositions and novel use in particular as C-MET inhibitors

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
ERNEST LACEY，等: "STRUCTURE-ACTIVITY RELATIONSHIPS OF BENZIMIDAZOLE CARBAMATES AS INHIBITORS OF MAMMALIAN TUBULIN, IN VITRO", 《BIOCHEMICAL PHARMACOLOGY》 *
JOSEPH E. DRUMM，等: "Facile preparation of fused ring azolylureas", 《TETRAHEDRON LETTERS》 *
PAUL S. CHARIFSON，等: "Novel Dual-Targeting Benzimidazole Urea Inhibitors of DNA Gyrase and Topoisomerase IV Possessing Potent Antibacterial Activity: Intelligent Design and Evolution through the Judicious Use of Structure-Guided Design and Stucture-Activity Relationships", 《J. MED. CHEM.》 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20220051569A (en) * 2020-10-19 2022-04-26 고려대학교 산학협력단 Benzimidazole derivatives or its pharmaceutically acceptable salts and use thereof
KR102516896B1 (en) * 2020-10-19 2023-04-03 고려대학교 산학협력단 Benzimidazole derivatives or its pharmaceutically acceptable salts and use thereof
CN115594667A (en) * 2022-10-18 2023-01-13 北京京佑奇康科技有限公司（Cn） Benzimidazole compound and preparation method and application thereof
CN115594667B (en) * 2022-10-18 2024-05-10 北京京佑奇康科技有限公司 Benzimidazole compound, and preparation method and application thereof

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