Withanone from Withania somnifera Attenuates SARS-CoV-2 RBD and Host ACE2 Interactions to Rescue Spike Protein Induced Pathologies in Humanized Zebrafish Model

Abstract

SARS-CoV-2 engages human ACE2 through its spike (S) protein receptor binding domain (RBD) to enter the host cell. Recent computational studies have reported that withanone and withaferin A, phytochemicals found in Withania somnifera, target viral main protease (M Pro) and host transmembrane TMPRSS2, and glucose related protein 78 (GRP78), respectively, implicating their potential as viral entry inhibitors. Absence of specific treatment against SARS-CoV-2 infection has encouraged exploration of phytochemicals as potential antivirals. Aim: This study aimed at in silico exploration, along with in vitro and in vivo validation of antiviral efficacy of the phytochemical withanone. Methods: Through molecular docking, molecular dynamic (MD) simulation and electrostatic energy calculation the plausible biochemical interactions between withanone and the ACE2-RBD complex were investigated. These in silico observations were biochemically validated by ELISA-based assays. Withanone-enriched extract from W. somnifera was tested for its ability to ameliorate clinically relevant pathological features, modelled in humanized zebrafish through SARS-CoV-2 recombinant spike (S) protein induction. Results: Withanone bound efficiently at the interacting interface of the ACE2-RBD complex and destabilized it energetically. The electrostatic component of binding free energies of the complex was significantly decreased. The two intrachain salt bridge interactions (K31-E35) and the interchain long-range ion-pair (K31-E484), at the ACE2-RBD interface were completely abolished by withanone, in the 50 ns simulation. In vitro binding assay experimentally validated that withanone efficiently inhibited (IC 50 =0.33 ng/mL) the interaction between ACE2 and RBD, in a dose-dependent manner. A withanone-enriched extract, without any co-extracted withaferin A, was prepared from W. somnifera leaves. This enriched extract was found to be efficient in ameliorating human-like pathological responses induced in humanized zebrafish by SARS-CoV-2 recombinant spike (S) protein. Conclusion: In conclusion, this study provided experimental validation for computational insight into the potential of withanone as a potent inhibitor of SARS-CoV-2 coronavirus entry into the host cells.

Identification of Novel Gene Signatures using Next-Generation Sequencing Data from COVID-19 Infection Models: Focus on Neuro-COVID and Potential Therapeutics

“… David et al, 2021 in their MedRixv preprint showed that a common variant of TMPRSS2 protects against COVID-19. In silico screening of several phytochemicals identified that Withanone, one of the constituents of ashwagandha, showed a potential inhibition of ACE2 ( Balkrishna et al, 2021 ). Additionally, Ghosh et al (2021) used molecular dynamic simulations and pharmacophore modeling approaches to predict the highly potent small-molecule derivative of withaferin A that potentially inhibits SARS-CoV-2 protease (Mpro), a favorable future therapeutic against COVID-19.…”

Influence of Patient-Reported Treatment Satisfaction on Psychological Health and Quality of Life Among Patients Receiving Divya-Swasari-Coronil-Kit Against COVID-19: Findings from a Cross-Sectional “SATISFACTION COVID” Survey

“…The stress subscale was assessed from questions 1, 6, 8, 11, 12, 14, and 18. The total score of stress subscale was subdivided into normal (0−10), mild (11)(12)(13)(14)(15)(16)(17)(18), moderate (19)(20)(21)(22)(23)(24)(25)(26), severe (27)(28)(29)(30)(31)(32)(33)(34), and extremely severe stress (35)(36)(37)(38)(39)(40)(41)(42). The DASS-21 has been demonstrated as valid and reliable for assessing mental health in previous COVID-19 related researches in the Indian population.…”

“…These formulations have been shown to suppress inflammatory storm, modulate immune function and reduce lung injury caused by SARS-CoV-2 S-protein under in-vitro and in-vivo experimental conditions. [12][13][14][15] The fundamental research and clinical study with its constituents showed that Divya-Swasari-Coronil-Kit promising bidirectional regulation in the symptoms associated with SARS-CoV-2 infection. Results from a doubleblind, randomized, controlled trial showed expedite virological clearance and concomitantly reduce the risk of viral dissemination.…”

Free Energy Landscapes and Residue Network Analysis for Six SARS-CoV-2 Targets in Complex with Plausible Phytochemical Inhibitors from Withania Somnifera: 1 µS Molecular Dynamics Simulations

“…Further, withanolides from Withania somnifera, showed anti-viral activity for Human Papilloma Virus, influenza viruses, herpes simplex virus, sindbis virus, parainfuenze-3 virus, human cytomegalovirus, and dengue virus type-2 (Amoros et al, 1994;Cai et al, 2015;Kwon et al, 2020;Latheef et al, 2017;Lyu et al, 2005;Munagala et al, 2011;Serkedjieva et al, 1992;Zandi et al, 2011). Withanone from Withania somnifera was also observed to interact with main protease (Mpro) and spike protein of SARS-CoV-2 (Kumar et al, 2020;Tripathi et al, 2020;Balkrishna et al, 2021).…”