Design,Systhesis And Biol Ogical Evaluation Of Flavonoid Derivatives As Inhibitors Of Tubulin And HIF-1?

Objective: Designing and synthesizing a flavonoid compound that can inhibit tumor vascularity and inhibit glycolysis of tumor cells by inhibiting tubulin and HIF-1?,so as to achieve excellent antitumor activity.Methods: The 3',4',5'-trimethoxy-7-OH-flavonoids were obtained through four steps using resorcinol,chloroacetonitrile and 3,4,5-trimethoxybenzaldehyde as raw materials.A series of benzimidazole derivatives were connected with 1,2-dibromoethane,1,3-dibromopropane,and 1,4-dibromobutane,respectively,and the products were purified by silica gel column chromatography to obtain 24 novel 3',4',5'-trimethoxyflavonoid benzimidazole derivatives.Their structures were characterized by 1H NMR,13 C NMR,and MS.The anti-proliferative activities of those 3',4',5'-trimethoxyflavonoid benzimidazole derivatives were tested by MTT colorimetric assay on human gastric cancer cells(MGC-803),human breast cancer cells(MCF-7),human hepatoma cells(Hep G-2)and murine gastric cancer cells(MFC).The effects of compound 15 on the apoptosis and cell cycle of MFC cells were determined by flow cytometry.In vivo antitumor activity of compound 15 was achieved in gastric-bearing mice and gastric-bearing nude mice.Solid tumors of tumor-bearing nude mice were sectioned and treated with a kit for detection of HIF-1? and tubulin to determine the effect of compound 15 on the expression of HIF-1? and tubulin.The computer-assisted drug design software SYBYL was used to simulate the binding ability of compound 15 to tubulin and give a fitting score.Results: A series of 3',4',5'-trimethoxyflavone benzimidazole derivatives were successfully synthesized and identified.In vitro antiproliferation assay results showed compound 15 namely 7-(3-(2-chloro-1H-benzo[d]imidazol-1-yl)-pro-poxy)-2-(3,4,5-trimethoxy ethane)Phenylphenyl)-4H-chromene-4-one exhibits significant antiproliferative activity against the above four tumor cells with IC50 values of 20.47 ± 2.07,43.42 ± 3.56,35.45 ± 2.03 ?M,and 23.47 ± 3.59 ?M,respectively.The results of the effects of compound 15 on cell cycle and apoptosis indicated that compound 15 was able to arrest MFC cell cycle at G1 phase and promoted cell apoptosis.Both effects were dose-dependent.In vivo anti-tumor results showed that compound 15 exhibited significant anti-tumor activity,while there was no significant decrease in body weight compared to saline group.Immunohistochemistry results showed that compound 15 can inhibit the expression of HIF-1? and tubulin.The results of molecular docking showed that compound 15 can form hydrogen bonds with the amino acid residues LYS-25 and THR-179 of tubulin,indicating that compound 15 has a certain binding ability to tubulin.Conclusion: Compound 15,namely 7-(3-(2-chloro-1H-benzo[d]imidazol-1-yl)propoxy)-2-(3,4,5-trimethoxyphenyl)-4H-Chrzen-4-one is a potential antitumor active compound.