HR-LCMS-guided metabolomics and in silico prioritization of antisnake venom agents from Cissus repanda: A sustainable discovery approach

Abstract: Snakebite envenomation is a neglected tropical disease with a substantial public health burden, particularly in Africa and Asia. Among venom components, phospholipase A2 (PLA2) enzymes play a central role in venom-induced tissue damage and inflammation. In this study, we report a non-animal, sustainability-aligned workflow for evaluating the antisnake venom potential of the methanol extract of Cissus repanda using enzyme-based in vitro assays, HR-LCMS-guided metabolomic profiling, and in silico analysis. The methanol extract was assessed for inhibitory activity against PLA2 enzymes from Naja nigricollis and Echis ocellatus venoms using an in vitro acidimetric assay, avoiding the use of animal models and reducing biological resource burden. The extract demonstrated significant PLA2 inhibition, with maximum and minimum inhibition values of 85.4% and 42.7% at concentrations of 1.25 and 10.0 mg/mL, respectively. High-resolution liquid chromatography–mass spectrometry (HR-LCMS) enabled efficient identification of eleven putative bioactive metabolites. To further minimize experimental redundancy and chemical waste, these compounds were prioritized using molecular docking, yielding binding affinities ranging from −6.8 to −5.2 kcal/mol. Three compounds with the most favorable binding poses were selected for post-docking analyses, and in silico ADMET evaluation to assess pharmacokinetic and safety-related properties. This integrated approach demonstrated how HR-LCMS-guided metabolomics and computational screening can reduce solvent use, experimental waste, and ethical burden in natural product–based antivenom research. The findings support Cissus repanda as a promising renewable source of antisnake venom leads and highlight a green chemistry–oriented framework for sustainable bioactive compound discovery