Paclitaxel (PTX), a cytotoxic drug, primarily acts on microtubules. At high concentrations, Cell cycle arrest is primarily caused during the G2/M phase. polyunsaturated fatty acid gamma linolenic acid (GLA), shows cytotoxic effects on different cancer cell types. In combination, it synergistically enhanced anti tumorigenic activity of chemotherapeutic agents. This investigation is focused on the formulation, optimization, and characterization of PTX-GLA liposomes and their efficacy in combating breast cancer. GLA primarily replaces lipid constituents within liposomes to enhance the anticancer efficacy of PTX-GLA liposome. It was prepared using the thin film hydration method and optimized through Box-Behnken design employing Design-Expert software. The PTX-GLA liposome was characterised on the basis of its size, shape, and surface characteristics using zeta potential analysis, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Fourier-transform infrared (FTIR) spectroscopy confirmed the compatibility of PTX-GLA-liposomes. The in-vitro drug release tests demonstrated that the PTX-GLA liposome exhibited sustained and regulated release kinetics over a duration of 48 hours, while maintaining excellent stability. The IC50 values of PTX liposomes, GLA liposomes, and PTX-GLA liposomes against MCF-7 cells were determined to be IC50 11.61 µM,12.31 µM and 11.21 µM, respectively. Cellular uptake investigations on MDA MB-231 cells demonstrated enhanced internalization of PTX-GLA liposomes compared to GLA liposomes. Additionally, the anti inflammatory properties of GLA may mitigate specific adverse effects associated with chemotherapy. The synergistic combination of GLA and Paclitaxel may enhance the overall efficacy of treatment through mechanisms such as anti inflammatory and pro-apoptotic actions.