Pharmacodynamices And Mechanism Of Novelantitumor Tetrazanbigen Derivative T-1019

ObjectThe purpose of this study is to explore the in vitro antitumor activities and mechanism of T-1019Methods1.Crystal violet staining,colony formation assay and CCK-8 assay were used to detect the anti-proliferation effects of T-1019 on Hep G2 and A549 cell lines.2.T-1019-induced cell apoptosis was measured by flow cytometry assay.3.The effects of T-1019 on PPARγand its related signaling pathways were detected by transactivation assay,TR-FRET binding assay and western blot（WB）assay.4.Oil red O staining was utilized to explore the effect of T-1019 on lipid droplets accumulation.5.The relationship between anti-tumor effect of T-1019 and PPARγwas investigated by combined with PPARγagonist or antagonist.6.Finally,the binding patterns of T-1019,TNBG with PPARγ-LBD were predicted through molecular docking.Results1.T-1019 strongly inhibited proliferation of Hep G2 and A549,and the IC₅₀values were 0.70μM and 0.55μM respectively.2.T-1019 could induce apoptosis of Hep G2 and A549.3.According to WB results,T-1019 could upregulate the levels of PPARγand PTEN,downregulate the phosphorylation of AKT4.Oil red O staining results demonstrated that T-1019 could induce lipid droplets accumulation.5.The antitumor activity of T-1019 could be reversed by PPARγinhibitor T0070907.This result suggested the antitumor effect of T-1019may be mediated by PPARγ.6.Results of Transactivation assay and TR-FRET binding assay suggested that T-1019 is a potential PPARγagonist.7.Molecular docking demonstrated that T-1019 bound to PPARγ-LBD in a similar mode to leading compound TNBG and positive control PPARγpartial agonist GSK1997132B.ConclusionIn this work,T-1019 was found to inhibit the proliferation in Hep G2and A549.It is a potential PPARγpartial agonist with antitumor activity.