Objective <p>Breast cancer is the most frequently diagnosed cancer among women, posing significant health risks. This study investigates niosome nanoparticles as a delivery system for Cyclophosphamide (CYC) and Sodium Oxamate (SO) to target apoptotic pathways in MDA-MB-231 breast cancer cells.</p> Methods <p>Lipid-based niosomes were prepared using the thin-film hydration method and characterized for size, zeta potential, and Fourier-transform infrared spectroscopy (FTIR) profiles. MDA-MB-231 cells were treated with CYC and SO-loaded niosomes, and cell viability was assessed using the MTT assay. Flow cytometry with annexin V/PI labeling evaluated apoptosis, while real-time PCR and Western blotting analyzed levels of NRF2 and key apoptotic proteins.</p> Results <p>Niosomes exhibited favorable properties with a mean particle size of 87.98 nm and a zeta potential of − 7.44 mV. Treatment significantly reduced cell viability compared to controls, indicating cytotoxic effects. Flow cytometric analysis revealed a substantial increase in apoptotic cells post-treatment. Western blot analysis showed elevated NRF2 protein levels and increased expression of caspase-3 and Bax, confirming activation of apoptotic pathways.</p> Conclusions <p>The co-delivery of CYC and SO via niosomes effectively targets apoptotic pathways in MDA-MB-231 breast cancer cells. The enhanced cytotoxicity and induction of apoptosis suggest that this drug delivery system may serve as an effective strategy for managing breast cancer.</p> Graphical Abstract <p></p>