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

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.