Identification of Gallagyldilacton as a Mutation-Biased EGFR Inhibitor from Punica granatum Peel via Integrative Computational Analysis

Abstract Non-small cell lung cancer (NSCLC) accounts for the majority of lung cancer cases worldwide and remains a leading cause of cancer-related mortality. Aberrant activation of the epidermal growth factor receptor (EGFR), driven by overexpression or mutation, represents a central oncogenic mechanism in NSCLC and a primary target of tyrosine kinase inhibitors (TKIs). Despite substantial clinical success, the emergence of resistance-conferring EGFR mutations continues to limit the long-term efficacy of existing therapies. In this study, we employed an integrative in silico framework combining network pharmacology, molecular docking, molecular dynamics simulations, and MM-PBSA analyses to investigate the therapeutic potential of phytochemicals derived from Punica granatum peel against NSCLC-associated molecular targets. Network analysis identified EGFR, AKT1, and matrix metalloproteinases (MMPs) as key nodes within NSCLC-related signaling pathways. Among the screened compounds, gallagyldilacton and naringenin emerged as prominent candidates with distinct target interaction profiles. Gallagyldilacton exhibited computationally stable and energetically favorable binding to multiple mutant EGFR variants, including T790M and L858R, while displaying comparatively weaker affinity toward wild-type EGFR, consistent with a mutation-biased inhibitory profile. In contrast, naringenin demonstrated preferential interactions with AKT1 and select MMPs, consistent with potential modulation of proliferative and metastatic signaling pathways. Comparative analyses with approved TKIs revealed that gallagyldilacton achieves binding stability and interaction patterns comparable to established inhibitors for specific EGFR mutants. Collectively, these findings support the potential of Punica granatum peel phytochemicals as sources of selective, multitarget modulators in NSCLC and highlight gallagyldilacton as a compound exhibiting mutation-biased EGFR interactions and a candidate for further experimental investigation.