Open Access

ARTICLE

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

HASSAN M. OTIFI¹, MISHARI ALSHYARBA², MAJED AL FAYI^3,4, AYED A. DERA^3,4, PRASANNA RAJAGOPALAN^3,4,*

1 Department of Pathology, College of Medicine, King Khalid University, Abha, Saudi Arabia
2 Department of Surgery, College of Medicine, King Khalid University, Abha, Saudi Arabia
3 Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Khalid University, Abha, Saudi Arabia
4 Central Research Laboratory, College of Applied Medical Sciences, King Khalid University, Abha, Saudi Arabia

* Corresponding Author: Prasanna Rajagopalan,

Oncology Research 2021, 29(3), 217-227. https://doi.org/10.32604/or.2022.03570

Received 26 April 2022; Accepted 07 July 2022; Issue published 01 August 2022

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 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 G₀/G₁, G₂M 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.

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