LC-MS Profiling and Anti-Inflammatory Activity of Saurauia fasciculata Bark: Network Pharmacology, Molecular Docking, MD Simulation, and in-Vitro Validation

Abstract

Background: Saurauia fasciculata has been traditionally utilized in Nepalese medicine for various therapeutic applications. This study aimed to characterize the phytochemical profile and evaluate the anti-inflammatory potential of S. fasciculata bark extract using an integrated in-silico and in-vitro approach. Methods: Methanolic bark extract was prepared by sequential maceration and analyzed using Fourier transform infrared spectroscopy (FT-IR) for functional group characterization and liquid chromatography–mass spectrometry (LC–MS/MS) for phytochemical profiling. Network pharmacology analysis was performed to predict inflammation-related targets. Key protein targets (PIK3R1 and PIK3CA) were subjected to molecular docking (AutoDock Vina v1.2.0) with identified phytoconstituents, followed by 100 ns molecular dynamics simulations using Schrödinger Desmond (release 2022-3). Post-simulation analysis was performed to validate the complex stability. Pharmacokinetic properties of the top phytoconstituents were evaluated in silico. Anti-inflammatory potential of the crude extract was validated in vitro using the protein denaturation assay, with aspirin as the standard. Results: LC–MS analysis revealed multiple bioactive constituents, including Equol, 2′-methoxy-6,8-dimethyl flavone, Quercetin, Retusin, and Corosolic acid. Network pharmacology highlighted PIK3R1 and PIK3CA as pivotal inflammation-related targets. Molecular docking showed that Equol had the highest binding affinity toward PIK3-δ (6PYR; −9.0 kcal/mol) and PIK3-α (8EXL; −9.1 kcal/mol). MD simulations confirmed stable protein–ligand interactions, and PCA and MM/GBSA analyses supported strong binding stability and energetics. In vitro, the extract inhibited protein denaturation in a dose-dependent manner, with an IC50of 112.10 ± 0.57 µg/mL compared to 75.33 ± 0.73 µg/mL for aspirin, suggesting moderate anti-inflammatory activity. Conclusion: This study provides preliminary evidence supporting the anti-inflammatory potential of S. fasciculata bark extract, with Equol identified as a promising candidate for further investigation.