Preprint / Version 1

Withania somnifera extracts induced attenuation of HIV-1: a mechanistic approach to restrict viral infection

Authors

  • Pratiksha Jadaun ICMR-National AIDS Research Institute, Pune, 411026 MH India
  • R Harshithkumar ICMR-National AIDS Research Institute, Pune, 411026 MH India
  • Shraddha Gaikwad ICMR-National AIDS Research Institute, Pune, 411026 MH India
  • Chandrabhan Seniya School of Biosciences, Engineering and Technology, VIT Bhopal University, Bhopal, 466114 MP India
  • Swapnil Borse CCIH-Interdisciplinary School of Health Sciences, AYUSH-Center of Excellence, Savitribai Phule Pune University, Pune, 411007 MH India
  • Ashish Gawai Anuradha College of Pharmacy, Buldana, 443201 MH India
  • Preeti Chavan-Gautam CCIH-Interdisciplinary School of Health Sciences, AYUSH-Center of Excellence, Savitribai Phule Pune University, Pune, 411007 MH India
  • Girish Tillu CCIH-Interdisciplinary School of Health Sciences, AYUSH-Center of Excellence, Savitribai Phule Pune University, Pune, 411007 MH India
  • Anupam Mukherjee ICMR-National AIDS Research Institute, Pune, 411026 MH India

Keywords:

Anti-HIV-1, Withania somnifera, Ashwagandha, Ashwagandhanolide, Withanolide, Withacoagin, Withaferin, Molecular docking

Abstract

Several anti-retroviral drugs are available against Human immunodeficiency virus type-1, but have multiple adverse side effects. Hence, there is an incessant compulsion for effectual anti-retroviral agents with minimal or no intricacy. Traditionally, natural products have been the most successful source for the development of new medications. Withania somnifera, also known as Ashwagandha, is the utmost treasured medicinal plant used in Ayurveda, which holds the potential to give adaptogenic, immunomodulatory, and antiviral effects. However, its effect on HIV-1 replication at the cellular level has never been explored. Herein, we focused on the anti-HIV-1 activity and the probable mechanism of action of hydroalcoholic and aqueous extracts of Withania somnifera roots and its phytomolecules. Methods The cytotoxicity of the extracts was determined through MTT assay, while the in vitro anti-HIV-1 activity was assessed in TZM-bl cells against the HIV-1 strains of X4 and R5 subtypes. Results were confirmed in peripheral blood mononuclear cells, using the HIV-1 p24 antigen assay. Additionally, the mechanism of action was determined through the Time of Addition assay, which was further validated through the series of enzymatic assays, i.e. HIV-1 Integrase, Reverse transcriptase, and Protease assays. To explore the role of the identified active metabolites of Withania somnifera in antiretroviral activity, molecular docking analyses were performed against these key HIV-1 replication enzymes. Results The hydroalcoholic and aqueous extracts of Withania somnifera roots were found to be safer at the sub-cytotoxic concentrations and exhibited their ability to inhibit replication of two primary isolates of HIV-1 through cell-associated and cell-free assays, in dose-dependent kinetics. Several active phytomolecules found in Withania somnifera successfully established hydrogens bonds in the active binding pocket site residues responsible for the catalytic activity of HIV replication and therefore, signifying their role in the attenuation of HIV-1 infection as implied through the in silico molecular docking studies. Conclusions Our research identified both the hydroalcoholic and aqueous extracts of Withania somnifera roots as potent inhibitors of HIV-1 infection. The in silico analyses also indicated the key components of Withania somnifera with the highest binding affinity against the HIV-1 Integrase by 12-Deoxywithastramonolide and 27-Hydroxywithanone, HIV-1 Protease by Ashwagandhanolide and Withacoagin, and HIV-1 Reverse transcriptase by Ashwagandhanolide and Withanolide B, thereby showing possible mechanisms of HIV-1 extenuation. Overall, this study classified the role of Withania somnifera extracts and their active compounds as potential agents against HIV-1 infection. Supplementary Information The online version contains supplementary material available at 10.1186/s12985-023-02130-y. Keywords: Anti-HIV-1, Withania somnifera, Ashwagandha, Ashwagandhanolide, Withanolide, Withacoagin, Withaferin, Molecular docking

Downloads