Cancer remains a major health challenge worldwide, with breast cancer being one of the most frequently diagnosed malignancies. Conventional chemotherapy is often associated with severe adverse effects and poor selectivity, which limits its clinical success. Herbal medicines have gained increasing attention due to their safety and therapeutic potential. Withania somnifera is a well-known medicinal plant possessing significant anticancer activity attributed to the presence of withanolides. However, its application is restricted by poor solubility and bioavailability. The present study was aimed to develop a PLGA-based nanoformulation of Withania somnifera extract and evaluate its apoptotic activity against MCF-7 breast cancer cell line. The nanoformulation was prepared using nanoprecipitation technique and characterized for particle size, zeta potential and entrapment efficiency. In-vitro cytotoxicity was evaluated using MTT assay. The nanoformulated extract exhibited enhanced cytotoxic and apoptotic activity compared to the crude extract. The findings suggest that PLGA-based nanoformulation of Withania somnifera may offer a potential nano-herbal strategy for further investigation in breast cancer management.