Isolation and structure elucidation of a steroidal moiety from Withania somnifera and in silico evaluation of antimalarial efficacy against artemisinin resistance Plasmodium falciparum kelch 13 protein

Abstract

According to the 2021 Malaria report, 241 million clinical episodes with 627000 deaths penalty was estimated across the worldwide. However, mutation in the propeller domain of Plasmodium falciparum kelch 13 protein resulted in longer parasite clearance time following an artemisinin-based treatment and had a greater survival rate of ring-stage parasites even after a brief exposure to a high dose of artesunate. Clinical manifestations become more complex and worse with the emerging trend of drug resistance against artemisinin derivatives and the poor effectiveness of malaria vaccination drive. Steroidal lactone (withanolide) moiety (C-28) isolated from methanolic leaf extract Withania somnifera show a greater affinity towards Pfkelch 13 protein in comparison to the artemisinin derivatives (artesunate, artemether). The isolated compound was characterized to be withaferin A with a percentage yield of 29.01% w/w in chloroform fraction, 1.75% w/w in methanolic extract, and 0.29% w/w in raw leaf powder. Structure-based analysis shows that withaferin A (docking score −8.253, −9.802) has a higher affinity for two distinct binding pockets I and II of the Plasmodium falciparum kelch 13 protein than artesunate (docking score −4.470, −3.656). Further, Gibbs binding free energy signifies thermodynamic stability of the docked complex of withaferin A (-43.25, −43.76 Kcal/mol) in comparison to artesunate docked complex (-8.49, −5.75 Kcal/mol). The pharmacokinetic profile of withaferin A shows more drug-likeness characteristics without violating Jorgensen’s rule of three, and Lipinski's rule of five. Hence above experimental findings suggest withaferin A could be a suitable therapeutic adjunct for preclinical evaluation of antimalarial potentiality in artemisinin-resistant malaria.

HIGHLIGHTs

• Malaria is a life-threatening parasitic disease caused by Plasmodium species.

• The emerging trend of artemisinin resistance and severe side effects (CNS and cardiotoxicity) are the potential challenges faced by antimalarial therapeutics.

• Artemisinin-mimic potentiality (ROS-mediated antiparasitic activity) of withaferin A shows a strong affinity towards artemisinin resistance Plasmodium falciparum kelch 13 protein.

• The pharmacokinetic profiling of the withaferin A signifies its drug-likeness characteristics without violating Jorgensen’s rule of three, and Lipinski's rule of five.

• Based on molecular docking and pharmacokinetic profiling, withaferin A could be a suitable therapeutic adjunct for preclinical investigation of antimalarial potentiality in artemisinin-resistant malaria.

Communicated by Ramaswamy H. Sarma

Graphical Abstract

Keywords:

• Withania somnifera
• isolation
• withaferin A
• Plasmodium falciparum kelch 13 protein
• molecular docking

Acknowledgments

The authors acknowledge the help of the Department of Biotechnology (DBT), Govt. India. The authors also thank Dr. Prajwal Nandekar, Senior Scientist II, Schrödinger, for providing the technical training on the Schrodinger suit.

Disclosure statement

The authors declare no conflict of interest.

Additional information

Funding

The research was funded by the DBT Builder project with order no BT/INF/22/SP45078/2022.