Targeting Yes-Associated Protein (YAP) in Breast Cancer: In Silico Molecular Dynamics, Luminescence-Based In Vitro, and In Vivo Validation of Rauvolfia tetraphylla-Derived Inhibitors

The study aims to elucidate the pharmacological mechanism of Rauvolfia tetraphylla against breast cancer through a comprehensive, multi-faceted approach. This includes molecular docking, molecular dynamics, and experimental validation. Initial screening via ADME analysis and network pharmacology identified key compounds and potential targets. Protein-protein interaction (PPI) network analysis pinpointed Yes-associated protein-1 (YAP) as a crucial target. Molecular docking revealed that three compounds-ajmaline, reserpine, and serpentine-exhibited strong binding affinities with YAP, with scores of -6.5 to -6.7 kcal/mol. Molecular dynamics simulations were conducted to assess the stability of these interactions further. Experimental validation showed R. tetraphylla inhibited breast cancer cell proliferation, with an IC50 of 348.69 μg/mL, while demonstrating cytoprotective effects on Vero cells (IC50: 1056.23 μg/mL). Migration assays indicated an 88.5% reduction in cell migration, and increased ROS levels signaled elevated stress in cancer cells. Apoptosis was confirmed by AO/EtBr staining. In vivo validation in a DMBA-induced mouse model confirmed significant tumor growth inhibition, supported by changes in YAP expression and histopathological analysis. These findings highlight R. tetraphylla as a promising therapeutic candidate against breast cancer, offering insights into its mechanisms and potential for future drug development and clinical applications.