Design,Synthesis And Anti-tumor Activity Of Flavone Derivatives Inhibiting Tumor Vascular And Glycolysis

Objective Designing and synthesizing a flavone derivative that can disrupt tumor vascular and inhibit glycolysis,so as to achieve excellent anti-tumor activity.Methods 1.Synthesis and characterization of target compounds:The3’,4’,5’-trimethoxy-7-hydroxyflavone,3’,4’,5’-trimethoxy-5,7-dihydroxyflavoneor5,6,7-trimethoxy-4’-hydroxyflavone were obtained using resorcinol,phloroglucinol or 3,4,5-trimethoxyphenol as raw materials.Four series of flavone derivatives were synthesized from fully synthetic flavonoids or chrysin with salicylic acid derivatives or benzimidazole derivatives.All the desired products were purified by silica gel column chromatography and confirmed by ¹H NMR and/or ¹³C NMR and/or MS.2.Anti-tumor activity evaluation of the target compound in vitro:The anti-proliferative activity of the target compound against different tumor cell lines and the cytotoxicity to normal cells were determined by MTT colorimetry.The compounds 7g,10v,18 and 5h were screened out to further determine its anti-tumor activity.Cell colony assay and wound healing assay were used to detect the effects of compounds on single cell proliferation and cell migration.Flow cytometry was used to detect the effect of compounds on tumor cell apoptosis and cycle.The effects of compounds 7g and 10v on the expression ofβ-tubulin and glycolytic proteins HIF-1α,HK-2 and PFK were detected by western blot analysis.3.Evaluation of anti-tumor activity of compounds 7g and 18 in vivo:Establish a mouse model of murine gastric cancer xenograft or a nude mouse model of human gastric cancer xenografts,and establish Drug groups（high-dose group,medium--dose group,low-dose group）,saline group,and 5-Fu control group were established and administered by intraperitoneal injection.Tumor volumes and body weights were monitored every other day to examine the effect of compounds on tumor suppression and the normal growth weight of mice or nude mice.The tumors of human gastric cancer-bearing nude mice were sectioned,and HE staining and immunohistochemistry were used to detect the effects of compound 7g on the expression ofβ-tubulin and glycolytic proteins HIF-1α.Results 92 novel flavone derivatives were successfully synthesized and identified.Most of flavone derivatives have anti-tumor activity through MTT colorimetry.Among them,compounds 7g and 18 showed the best antitumor activity against human gastric cancer cell line MGC-803.Compound 10v exhibited the best antitumor activity against human colorectal cancer cell line HCT-116.Compounds 18 showed the best antitumor activity against murine gastric cancer cell line MFC,and compound 5h showed the best antitumor activity against human gastric cancer cell line HGC-27.What’s more,compounds 7g and 10v down-regulated the expression ofβ-tubulin and glycolytic related proteins HIF-1α,HK-2 and PFK.In addition,the in vivo anti-tumor activity evaluation results showed that the compound 7g showed significant anti-tumor activity in both tumor-bearing mice and tumor-bearing nude mice,while there was no significant decrease in body weight compared to the saline group.Compound 18 showed good anti-tumor activity in tumor-bearing mice but was more toxic.Conclusion The pharmacophore combination principle was an effective strategy to design new anti-tumor drug candidates.Flavone derivatives 7g,10v,18 and 5h are a promising anti-tumor agent candidate,deserving further optimization and evaluation.