Antitumor Activity Evaluation Of Podophyllotoxin And 5-FU Conjugate C069

Podophyllotoxin(PPT)shows strong cytotoxic activity against many tumor cell lines.Many anti-cancer drugs,such as VP-16,VM-26,etc,are been yielded through the Structural modifications to PPT.However,their therapeutic uses are often hindered by problems such as poor water-solubility and acquired drug-resistance.So our group want to develop and explore new compounds from natural products with efficient anticancer activities and low toxicity,we selected different linkers to combine DDPT and 5-FU to synthesize a series of novel hybrids.Compound(4′-O-demethyl-4-deoxypodophyllic 4-((6-(2-(5-fluorouracil-yl)acetamido)hexyl)amino)-4-oxobutanoate(C069)shows strong cytotoxic activity against four cancer cell lines(A549、HCT-8、HeLa、HepG2),in this study,the effects and mechanisms of C069 on anti-tumor,anti-metastasis and anti-angiogenesis were determined.We investigated the anti-tumor activities of C069 by MTT assay,western blot analysis,Transwell assay,CAM assay and ELISA,etc.The results suggested that C069 could inhibit the proliferation and colony forming ability of A549 cells,stop the cells in the G2/M phase;C069 could inhibit the migration and invasion of A549 cells and reduce the expression of MMP-9 and the phosphorylation process of ERK and P38,promote the expression of TIMP-1.These results suggested that C069 could make the the expression of MMP-9 and TIMP-1 abnormal may be through the inhibition of ERK and P38 phosphorylation,and then led the migration and invasion be restrained;C069 could inhibit the proliferation and tube formation of HUVECs.It also could disrupt the neovessel of HUVECs in vitro.In addition,C069 could reduce the neovascularization of CAM in vivo;VEGF,MMP-9,MMP-2 and the phosphorylation of β-catenin and Akt could be inhibited by C069,too.It is suggested that C069 could decrease the phosphorylation of Akt,thus the expression of MMP-9 and MMP-2 decrease and then prevent the tumor angiogenesis.In this project,we preliminary investigated the anti-tumor activity of C069,included the ability of anti-tumor proliferation,anti-tumor metastasis and anti-angiogenesis.These results may contribute to the exploitation and application of the derivatives of podophyllotoxin.