LC-HRMS-based phytochemical profiling of the ethanolic extract of Moringa oleifera seeds against toxic hepatotoxicity supported by in silico network design, molecular docking and simulation studies

Toxic hepatitis or drug-induced liver injury (DILI) has emerged as an escalating health concern over the past few years and is driven by the hepatic metabolism of xenobiotic compounds. The present study evaluated the hepatoprotective efficacy of Moringa oleifera seed extract (MOSE) using an integrated strategy that encompassed HRMS-based profiling, ADMET screening, Metascape-based network pharmacology, molecular docking via SMINA, and molecular dynamics (MD) simulations performed through GROMACS. LC-HRMS profiling identified 3,085 putative metabolites, of which 14 were selected based on their relative abundance. Rigorous ADMET screening resulted in three shortlisted compounds, namely, trans-3-indoleacrylic acid, (R)-prunasin, and 5-(4-morpholinyl)pyrazolo[1,5-a]quinazoline-3-carbonitrile, which were investigated in detail. Network pharmacology analysis revealed five topological hub genes, HRAS, PIK3CA, MMP9, LGALS3, and NFE2L2, as core regulators of extract-mediated protection against DILI. The identified targets from the similarity ensemble approach (SEA) were significantly enriched in essential biological pathways, including the apoptosis and chemical carcinogenesis-ROS pathways as revealed by Metascape. Molecular docking analysis with SMINA indicated strong binding efficacy, especially for (R)-prunasin toward HRAS (-9.20 kcal/mol). A 100 ns GROMACS MD simulation corroborated the robustness of the interaction stability under physiological conditions. These results suggest that Moringa oleifera seeds confer hepatoprotective effects through the coordinated modulation of multiple targets that support cell survival networks, activate antioxidants, and inhibit apoptosis. This study offers a molecular-level rationale for advancing standardized MOSE-based therapies for liver injury via integrative systems pharmacology approaches.