Computational docking and in vitro analysis identifies novel arylidene analogue FPMXY-14 against renal cancer cells by attenuating Akt

Abstract

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 GI 50 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 IC 50 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.

Targeting Prolyl 4-Hydroxylase Subunit Beta (P4HB) in Cancer: New Roads to Travel

“…Consistent with our pan-cancer analysis, our previous studies [8,27] found that P4HB expression was higher in tumor samples than in normal samples and P4HB downregulation could significantly inhibit the cell proliferation of six PRAD cell lines, including LNCap, C4-2, C4-2B, PC3, DU145 and 22RV1 cells. In addition, renal cancer is also a common urinary tumor which ranks the 9th most common type of cancer in males and accounts for 4.2% of novel diagnoses [28][29][30]. Several studies used bioinformatic analysis to indicate that P4HB was significantly associated with prognosis for KIRC and KIRP patients [31][32][33][34][35][36], which was consistent with our finding in this study.…”

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“…The assay was carried out as described elsewhere [21] using kit method as per the manufacturer's instructions. Brie y, HUVEC cells at a concentration of 1x10 5 were grown on the provided migration inserts until con uent.…”