Polyphenol-based therapeutics for glioblastoma: validation from In-vitro cell viability assay and integrated onco-omics computational analysis.

Glioblastoma (GBM) is one of the most aggressive brain tumors, with poor therapeutic outcomes due to its complex molecular profile. This study investigated the multi-target potential of three natural polyphenols-Ferulic acid, Morin, and Mangiferin-through an integrative computational strategy combining network pharmacology, onco-omics, molecular docking, molecular dynamics (MD) simulations, and density functional theory (DFT). Cross-referencing polyphenol-associated targets with GBM-related genes identified 13 common targets (e.g., PTGS2, EGFR, ESR1, MMP9). Protein-protein interaction analysis showed significant connectivity (p = 1.54 × 10⁻⁸), highlighting their relevance in GBM. Gene Ontology and KEGG enrichment revealed roles in proliferation, apoptosis, and migration, with enrichment in PI3K-Akt and MAPK signaling pathways. Molecular docking confirmed stable binding, with Mangiferin showing the strongest affinities: -11.0 kcal/mol (6ESM), -8.2 kcal/mol (5UGC), -7.5 kcal/mol (5UFW), and -9.1 kcal/mol (5IKT). MD simulations revealed the 5IKT-Mangiferin complex to be the most stable, with a favorable binding free energy (-32.0 ± 4.4 kcal/mol), while PCA and free energy landscapes supported reduced conformational variability. DFT results further supported favorable electronic properties. In vitro assays showed dose-dependent cytotoxicity, with IC₅₀ values of 9.43 µM (Morin), 4.65 µM (Mangiferin), and 6.22 µM (5-FU). Collectively, Mangiferin emerged as the lead candidate with superior binding stability and multi-target potential against GBM. This integrated framework provides mechanistic insights supporting polyphenol-based therapeutic development and warrants further in vivo validation.