Developing a formulation of Tinospora cordifolia, identifying its active components, and assessing its anticancer effects on breast cancer cell lines.

Tinospora cordifolia (Willd.) Miers is a medicinal plant recognised for its pharmacological potential. This work presents the development of an innovative nano-liposomal formulation and assesses its anticancer efficacy against breast cancer cell lines. A sustainable green extraction method was employed to isolate bioactive compounds from T. cordifolia, followed by the development of a nano-liposomal formulation. Particle size and morphology were assessed using field emission scanning electron microscopy (FESEM), revealing soft, globular vesicles with an average diameter of ~ 153 nm. GC-MS profiling identified 35 phytoconstituents subjected to molecular docking against topoisomerase IIα to predict anticancer potential. The biological activity of the formulation was validated through MTT assay for cell viability, scratch assay for cell migration, and apoptosis assays in MCF-7 and MDA MB 231 breast cancer cell lines. Immunocytochemistry was used to evaluate the expression of Bcl-2, cytochrome-C, and caspase-3. ROS generation was also quantified to confirm the mechanism of action. In silico analysis identified glucobrassicin as a potent topoisomerase IIα inhibitor (docking score: - 10.2655). The formulation exhibited dose-dependent cytotoxicity, inhibited cell migration, and induced apoptosis in both cell lines. ROS-mediated cell death was associated with increased cytochrome-C and caspase-3 expression and decreased Bcl-2 levels. This study underscores the value of integrating green nanotechnology, computational docking, and functional cell-based assays to identify and characterise the bioactive phytochemicals. The T. cordifolia-based liposomal formulation demonstrated promising anticancer activity and warrants further preclinical evaluation as a candidate for breast cancer therapy.