In silico to In vivo development of a polyherbal against Haemonchus contortus

Anu Rahal; D K Sharma; Ashok Kumar; Nitika Sharma; Deen Dayal

doi:10.1016/j.heliyon.2022.e08789

In silico to In vivo development of a polyherbal against Haemonchus contortus

Heliyon. 2022 Jan 19;8(1):e08789. doi: 10.1016/j.heliyon.2022.e08789. eCollection 2022 Jan.

Authors

Anu Rahal¹, D K Sharma¹, Ashok Kumar¹, Nitika Sharma¹, Deen Dayal¹

Affiliation

¹ Division of Animal Health, ICAR-Central Institute for Research on Goats, Makhdoom, Farah, Mathura, 281122, UP, India.

Abstract

Haemonchus contortus is a major constraint in the development of small ruminant subsector due to significant production losses incurred by it. The present study explores the antiparasitic potential of three anthelmintic plants (Butea monosperma, Vitex negundo and Catharanthus roseus (L.) G.Don) against H. contortus taking albendazole as the standard. In silico molecular docking and pharmacokinetic prediction studies were conducted with known bioactive molecules of these plants (palasonin, vinblastine, vincristine, betulinic acid and ursolic acid) against Glutamate Dehydrogenase (GDH) and tubulin molecules of the parasite. Methanolic extracts of these herbs were fractionated (hexane, ethyl acetate, chloroform and methanol) and used in in vitro larvicidal studies. Based on the in vitro data, two herbal prototypes were developed and clinically tested. All the 5 ligand molecules showed better binding affnity for GDH and tubulin protein as compared with albendazole and shared similar binding site in the core of the GDH hexamer with slight variations. Albendazole approximately stacked against GLY190A residue, showing hydrophobic interactions with PRO157A and a Pi-cation electrostatic interaction with ARG390 along with four hydrogen bonds. Vincristine formed 2 pi-anionic electrostatic bonds with ASP158 of B and C subunits alongwith hydrogen bonding and hydrophobic interaction and an additional pi-anion electrostatic interaction at ASP158A for vinblastine. Albendazole bound to α-tubulin next to colchicine site whereas vinblastine is bound at the nearby laulimalide/peloruside site of the dimer. Betulinic acid showed lateral interaction between the H2-H3 loop of one alpha subunit and H10 of the adjacent alpha subunit of two tubulin dimers. Ursolic acid and palasonin bound at the intradimer N site of microtubulin involving the H1-H7 and H1-H2 zone, respectively. The in vitro studies demonstrated good dose dependent anthelmintic potential. Both the prototypes were quite efficacious in clearing the infection, keeping it to a minimal for more than 5 months, probably, through direct anthelmintic effect through GDH, tubulin depolymerization and uncoupling as well as indirectly through immunomodulation along with antioxidant and anti-inflammatory properties.

Keywords: Betulinic acid; Butea monosperma (Lam.) Kuntze; Catharanthus roseus; Haemonchus contortus; Palasonin; Ursolic acid; Vinblastine; Vincristine; Vitex negundo Linn..