Phytoconstituents of Gmelina Arborea as Potential Breast Cancer Inhibitors: LC-MS/MS, Docking, MD Simulations, and In Vitro Evaluation.

Breast cancer remains one of the leading causes of cancer-related mortality in women, emphasizing the urgent need for safer and more effective therapeutic agents. Natural phytochemicals, due to their structural diversity and pharmacological potential, are increasingly recognized as promising candidates in anticancer research. In this study, the anticancer activity of the n-hexane extract from Gmelina arborea leaves was evaluated through a multidisciplinary approach involving LC-MS/MS analysis, molecular docking, molecular dynamics (MD) simulation, and in vitro cytotoxicity assays. LC-MS/MS profiling identified 83 phytochemicals based on accurate mass measurements and fragmentation patterns. Among them, hesperetin-3',5,7-tri-glucuronide, emerged as a key compound due to its potential interaction with the epidermal growth factor receptor (EGFR). Molecular docking studies using EGFR crystal structures (1M17 and 1IVO) revealed strong binding affinities of -9.122 and -7.246 kJ/mol, respectively. MD simulations further validated the complex stability for compound 77. Additionally, the extract demonstrated cytotoxic activity against MCF-7, HepG2, and HEK-293 cell lines, with the strongest activity observed in MCF-7 breast cancer cells. These findings were further supported with a clonogenic assay on MCF-7 cells using IC50 concentration. Overall, the findings highlight Gmelina arborea as a promising source of anticancer agents.