Integrated in vitro and in silico evaluation of the anti-inflammatory potential of Breynia vitis-idaea leaf extract using HR-LCMS profiling, network pharmacology and molecular docking study

This study investigates the anti-inflammatory potential of Breynia vitis-idaea (Phyllanthaceae) methanolic leaf extract using an integrated in silico and in vitro approach. HR-LCMS performed phytochemical profiling, followed by SwissADME predictions of ADME and drug-likeness. Potential molecular targets were identified by SwissTargetPrediction and analyzed by network pharmacology, protein-protein interaction (PPI), Gene ontology (GO), and KEGG pathway enrichment. Suggested ligand-target interactions were then evaluated through molecular docking and molecular dynamics (MD) simulation. Finally, anti-inflammatory activity was evaluated in vitro by egg albumin denaturation, bovine serum albumin (BSA) denaturation, and human red blood cell (HRBC) membrane stabilization assays. HR-LCMS identified 22 phytoconstituents, 07 of which exhibited good pharmacokinetic and drug-likeness features. Network pharmacology identified 381 shared targets between B. vitis-idaea phytoconstituents and inflammatory genes, with SRC emerging as the primary hub. Enrichment analyses (GO/KEGG) mapped SRC-associated targets to membrane rafts, where they govern oxidative stress and inflammation via kinase and transcriptional activities across TRP channel, Th17 differentiation, and AGE-RAGE signalling pathways. Molecular docking revealed strong SRC binding, peaking with 6-(2-Hydroxyethyl)-5,6-dihydrosanguinarine (−8.9 kcal/mol). A 100ns MD simulation confirmed this, demonstrating a highly stable SRC-ligand complex with persistent interactions and minimal active-site volatility. Validating these computational findings, the extract exhibited dose-dependent anti-inflammatory activity with a maximum inhibition of egg albumin and BSA denaturation as 68.6 ± 1.9% and 65.7 ± 2.1%, respectively, and 72.5±2.0% HRBC membrane stabilization at 200μg/mL. 6-(2-hydroxyethyl)-5,6-dihydrosanguinarine emerges as a highly promising anti-inflammatory lead targeting SRC-mediated pathways; however, advancing this candidate will require bioassay-guided isolation, detailed pharmacokinetic profiling, and in vivo validation.