Design,Synthesis And Anti-Tumor Activity Screening Of Novel 2-Aryl Benzimidazole Compounds

Objective:By analyzing the structure-activity relationship of compound CA-4,2-arylbenzimidazole derivatives containing CA-4 structure were designed and synthe sized,and their antitumor activities were studied in vitro.Methods:?1?benzimidaz ole ring was synthesized from 4-cl-5-methoxy and 3,4,5-trimethoxyacetaldehyde by methoxy substitution,iodation and cyclization.Two series of CA-4 derivatives were prepared by introducing arylacetylene group into Sonogashira reaction at 6-positio n.?2?Five kinds of tumor cells,A549,MCF-7,Caco-2,Eca-109 and SiHa,which are sensitive to CA-4,were selected.The inhibitory effect of the synthesized comp ounds on the proliferation of the four cell lines was determined by MTT method.?3?By using annexin v-apc/7-AAD apoptosis detection kit,cell cycle experiments and drug-induced apoptosis experiments were carried out on compound 6 and SiH a cells with the best inhibitory activity on tumor cells.?4?Western blot method w as used to study the inhibitory activity of intermediate compound 1 on PI3K signal ing pathway,and to analyze whether the compound can produce multi-target inhibit ion.Results:?1?sixteen benzimidazole derivatives were synthesized.Among them,compounds 1-8,5-methoxy-containing ring of benzimidazole,9-16,no 5-methoxy-co ntaining ring of benzimidazole,were all new compounds.The yield of the compou nds ranged from 24.7%to 83.6%.All the synthesized compounds were characterize d by 1H NMR and 13C NMR,and the new compounds were further confirmed by HRMS.?2?The compound has a certain value-added inhibition on five kinds of t umor cells,and has a strong anti proliferation effect on human cervical cancer SiH a cells and human breast cancer MCF-7 cells.Among them,the inhibitory activity of compound 1-8 on five tumor cells was significantly stronger than that of compo und 9-16,indicating that the presence or absence of methoxyl at the 5-position of benzimidazole had an important effect on the tumor proliferation inhibition of these compounds.The results of cell cycle experiments showed that when the concentra tion of compound 6 increased from 0.01 to 10?m,the percentage of cells in G2/M phase?cell aggregation rate?increased from 11.5 to 79.65%,which was dose-dependent.The experiment of promoting apoptosis showed that compound 6 could effectively induce SiHa cell apoptosis.When the concentration was 1?m,the prop ortion of cells in the pre and post apoptosis was 27.4%.When the concentration was 1?m,the proportion of cells in the pre and post apoptosis was 76.6%,whic h was concentration dependent.?3?Conclusion:16 benzimidazole derivatives were synthesized,16 of them were new compounds.The structure of the compound was confirmed by ¹H NMR,¹³C NMR and HRMS.The results of MTT assay showed that compound 6 and compound 1 had strong inhibitory effect on human lung canc er cell A549,human breast cancer cell MCF-7,human colon cancer cell Caco-2,h uman esophageal cancer cell Eca-109 and human cervical cancer cell SiHa in vitro,among which compound 6 had the strongest inhibitory activity on SiHa cell.Furt her cycle studies showed that compound 6 could block SiHa cells in G2/M phas e,and further apoptosis experiments showed that compound 6 had a good apoptosi s promoting effect on SiHa cells.