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 FAYI3,4, AYED A. DERA3,4, PRASANNA RAJAGOPALAN3,4,*
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 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
2M 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.
Keywords
Cite This Article
OTIFI, H. M., ALSHYARBA, M., FAYI, M. A., DERA, A. A., RAJAGOPALAN, P. (2021). Computational docking and
in vitro analysis identifies novel arylidene analogue FPMXY-14 against renal cancer cells by attenuating Akt.
Oncology Research, 29(3), 217–227.