Targeted therapies are gaining global attention to tackle Renal Cancer(RC).This study aims to screen FPMXY-14(novel arylidene analogue)for Akt inhibition by computational and in vitro methods.FPMXY-14 was subjected to...Targeted therapies are gaining global attention to tackle Renal Cancer(RC).This study aims to screen FPMXY-14(novel arylidene analogue)for Akt inhibition by computational and in vitro methods.FPMXY-14 was subjected to proton NMR analysis and Mass spectrum analysis.Vero,HEK-293,Caki-1,and A498 cell lines were used.Akt enzyme inhibition was studied with the fluorescent-based kit assay.Modeller 9.19,Schrodinger 2018-1,LigPrep module,and Glide docking were used in computational analysis.The nuclear status was assessed by PI/Hoechst-333258 staining,cell cycle,and apoptosis assays were performed using flow cytometry.Scratch wound and migrations assays were performed.Western blotting was applied to study key signalling proteins.FPMXY-14 selectively inhibited kidney cancer cell proliferation with GI50 values of 77.5 nM and 101.40 nM in Caki-1 cells and A-498 cells,respectively.The compound dose-dependently inhibited Akt enzyme with an IC50 value of 148.5 nM and bound efficiently at the allosteric pocking of the Akt when computationally analyzed.FPMXY-14 caused nuclear condensation/fragmentation,increased the sub G_(0)/G_(1),G_(2)M populations,and induced early,late phase apoptosis in both cells when compared to controls.Treatment of the compound inhibited wound healing and migration of tumor cells,while proteins like Bcl-2,Bax,and caspase 3 were also altered.FPMXY-14 effectively inhibited the phosphorylation of Akt in these cancer cells,while total Akt was unaltered.FPMXY-14 exhibited anti-proliferative and anti-metastatic activities in kidney cancer cells by attenuating the Akt enzyme.Further pre-clinical research on animals with a detailed pathway elucidation is recommended.展开更多
基金supported by the Deanship of Scientific Research at King Khalid University,Abha,Saudi Arabia,for funding this work through the General Research Project under Grant No. (G.R.P.1/39/39).
文摘Targeted therapies are gaining global attention to tackle Renal Cancer(RC).This study aims to screen FPMXY-14(novel arylidene analogue)for Akt inhibition by computational and in vitro methods.FPMXY-14 was subjected to proton NMR analysis and Mass spectrum analysis.Vero,HEK-293,Caki-1,and A498 cell lines were used.Akt enzyme inhibition was studied with the fluorescent-based kit assay.Modeller 9.19,Schrodinger 2018-1,LigPrep module,and Glide docking were used in computational analysis.The nuclear status was assessed by PI/Hoechst-333258 staining,cell cycle,and apoptosis assays were performed using flow cytometry.Scratch wound and migrations assays were performed.Western blotting was applied to study key signalling proteins.FPMXY-14 selectively inhibited kidney cancer cell proliferation with GI50 values of 77.5 nM and 101.40 nM in Caki-1 cells and A-498 cells,respectively.The compound dose-dependently inhibited Akt enzyme with an IC50 value of 148.5 nM and bound efficiently at the allosteric pocking of the Akt when computationally analyzed.FPMXY-14 caused nuclear condensation/fragmentation,increased the sub G_(0)/G_(1),G_(2)M populations,and induced early,late phase apoptosis in both cells when compared to controls.Treatment of the compound inhibited wound healing and migration of tumor cells,while proteins like Bcl-2,Bax,and caspase 3 were also altered.FPMXY-14 effectively inhibited the phosphorylation of Akt in these cancer cells,while total Akt was unaltered.FPMXY-14 exhibited anti-proliferative and anti-metastatic activities in kidney cancer cells by attenuating the Akt enzyme.Further pre-clinical research on animals with a detailed pathway elucidation is recommended.
