HR-LCMS guided Computational and Experimental Investigation of the Antihyperglycemic Potential of Dryopteris sparsa (L.) Sw. through phytochemical profiling, Network Pharmacology, Molecular docking and Molecular Dynamics Simulation

Dryoprteris sparsa (L.) Sw., a native fern to Eastern Himalayas has been used in traditional medicine; however, its antidiabetic potential and molecular mechanisms remain poorly understood. This study integrated computational and experimental approaches to investigate potential bioactive phytoconstituents and their putative multitarget mechanism relevant to type 2 diabetes mellitus (T2DM) Methods: The methanolic extract (FN4) of D. sparsa selected based on comparative in vitro activity was profiled using high-resolution liquid chromatography-mass spectrometry (HR-LCMS). Putatively annotated phytoconstituents were subjected to drug-likeness and ADME screening, followed by target prediction, network pharmacology, protein-protein interaction, Gene Ontology and KEGG pathway enrichment analyses. Molecular docking was perfomed against a-amylase, a glucosidase and interleukin-1b, followed by 100-ns molecular dynamics (MD) simulations of the prioritized compound. Experimental evaluation included enzyme inhibition assays and an oral glucose tolerance test (OGTT). Results: HR-LCMS putative annotated 31 phytoconstituents. Network pharmacology identified 839 overlapping predicted targets with IL1b, NFKB1, HIF1A, EGFR, CXCL8, ESR1, GNB1 and GNG2 emerging as hub proteins. Enrichment analyses implicated PI3K-Akt, NF-kB, AGE-RAGE, HIF-1, IL-17, Toll-like receptor and chemokine signaling pathways. Galangin was prioritized based on its favorable multi-target docking, pharmacokinetic and drug-likeness profiles and exhibited comparatively greater stability with ILβ during MD simulation. FN4 showed α-amylase (IC50=40.73±0.58μg/ml) and α-glucosidase (IC50= 36.68±0.07μg/ml) inhibition and produced a 20.69% reduction in glucose AUC at the higher dose during OGTT Conclusion: These finding support the antihyperglycemic potential of D. sparsa and identify galangin as a putative multitarget candidate for further investigation.