Design,Synthesis And Antitumor Activity Of B-ring Arylurea Flavonoid Derivatives

Objective: A series of B-ring aryl urea flavonoid derivatives were designed and synthesized.We hopes to screen out anti-tumor drug candidates that simulta-neously inhibit VEGF,glycolysis,and tubulin’s multiple targets.Methods: 1.Synthesis: Using 3,4,5-trimethoxyphenol and p-nitrobenzaldehyde as the starting materials for the synthesis,the important intermediate 5,6,7-trimethoxy-4’-nitroflavone was synthesized.The synthesized flavonoids are reacted with anilines with different substituents to obtain a series of novel arylurea flavonoid derivatives.Purify all target products by silica gel column chromatography,and use1 H NMR to determine the number of hydrogen atoms,carbon spectrum(13C NMR)to determine the number of carbon atoms,and high resolution mass spectrometry(HRMS)to determine The relative molecular mass of the compound.2.Activity evaluation: The anti-tumor activity was detected by the MTT colorimetric method.Anti-proliferative activities of five tumor cells including human liver cancer cells(Hep G-2),human triple-negative breast cancer cells(MDA-MB-231),human colon cancer cells(HCT-116),Human gastric cancer cells(HGC-27)and human breast cancer cells(MCF-7)were detected of all compounds.At the same time,we also tested the compounds against normal liver cells(L02)and human aortic vascular smooth muscle cells.The cytotoxicity of(HA-VSMC)and human gastric mucosal cells(GES-1),combined with the results of the compound’s toxicity tests on tumor cells and normal cells,selected compound 6r for further study.The cell colony experiment was used to examine the effect of compound 6r on cell colony formation.The effect of compound 6r on tubulin was detected by immunofluorescence experiment.Western blotting was used to detect the effect of compound6 r on the expression levels of VEGF and glycolysis-related proteins PKM2,PFKM and HK-2.3.Computer-aided drug design: Computer-aided drug design: Use SYBYL software to dock the compound and protein 4ASD,and calculate the corresponding score through a series of steps.And use Py MOL software to process the docking diagram of 6r.Use MOE software to calculate the molecular descriptor of the compound and evaluate the possibility of the compound becoming a medicine.Results: 20 novel B-ring aryl urea flavonoid derivatives were synthesized and identified.The in vitro anti-tumor activity results showed that most of the compounds have good anti-tumor activity.Among them,the compound 6r(1-(4-fluoro-3-trifluor-omethyl)-Phenyl)-3-[4-(5,6,7-trimethoxy-4-oxo-4H-chromium-2-yl)-phenyl]-urea)is the most prominent.Compound 6r down-regulated the expression of VEGF,PKM2,PFKM,and HK-2 protein in HGC-27 cells and Hep G-2 cells.Computer simulation results show that compound 6r may be a selective inhibitor of VEGF,but more research work is needed to increase the possibility of 6r being a drug.Conclusion: Compound 6r has a good anti-tumor performance and can be used as an anti-tumor drug for further research.