Bioactive Profiling and Evaluation of Anti-Proliferative Potential of Salvadora persica Bark Extract in Triple-Negative Breast Cancer Cells: An In Vitro and Computational Analysis

Background: Salvadora persica (S. persica), commonly known as ‘Miswak,’ has been used in ethnotraditional applications since ancient times. This study was formulated to examine bioactive phenolics and flavonoids from the hydroethanolic extract of S. persica bark, anticancer activity, and in silico binding interaction analysis with key therapeutic targets of triple-negative breast cancer (TNBC) cells. Methods: UHPLC was used to identify the phytochemicals in S. persica bark extract. Cell death was analyzed by MTT assay in TNBC MDA-MB-231 and MDA-MB-468 cells. Moreover, cellular apoptosis, ROS generation, MMP, and cell cycle checkpoints were also carried out. AutoDock Tools 1.5.7 and PyRx 0.8 tools were used for molecular binding interaction analysis. Results: Phytochemical analysis revealed the presence of total phenolic and total flavonoid content of 26.90 ± 0.46 μg GAE/mg and 54.51 ± 0.42 μg QE/mg of bark extract, respectively. UHPLC analysis confirmed the presence of fumaric acid, chlorogenic acid, rutin, and quercetin in the extract. S. persica significantly reduced cell viability of MDA-MB-231 and MDA-MB-468 cells with an IC50 value of 144 and 128 μg/mL, respectively. S. persica extract elevated ROS generation, loss of MMP, late apoptosis induction, and G2/M-phase cell cycle arrest, while it did not show any significant effect against normal kidney Vero cells. Molecular docking studies revealed that rutin showed strong binding affinity towards EGFR with B.E. = −9.8 and −9.5 Kcal/mol; FGFR1 with B.E. = −7.4 and −7.5 Kcal/mol; FGFR4 with B.E. = −7.5 and −7.9 Kcal/mol; and csGRP78 with B.E. = −9.0 and −9.3 Kcal/mol, using Autodock Vina and PyRx tools, respectively. SwissADME and drug-likeness analysis confirmed acceptable drug-like characteristics and favorable pharmacokinetic profiles of the identified molecules. Conclusions: This study highlighted the potential of phytochemicals from S. persica bark as promising compounds for the development of novel anticancer therapeutics.