The present study focused on the formulation, development, and characterization of a novel miconazole-based glycerosomal formulation to enhance topical antifungal drug delivery. Pre- formulation studies confirmed the purity and identity of miconazole through organoleptic properties, melting point analysis (176 °C), solubility profiling, UV spectrophotometry (λmax = 273 nm, R² = 0.9992), and FTIR analysis, which verified the presence of characteristic functional groups. Five glycerosomal formulations (F1–F5) were prepared and evaluated. All formulations exhibited stable organoleptic properties with no phase separation. Particle size ranged from 120.41 nm (F2) to 208.7 nm (F5), with F2 showing the smallest vesicles, while polydispersity index (18.6–32.6%) indicated uniform distribution, especially in F3. Zeta potential values suggested good stability, with F2 showing the highest negative potential (– 35.4 mV). Entrapment efficiency varied across formulations, with F2 demonstrating the highest drug loading at 96.52%. Scanning Electron Microscopy confirmed spherical, smooth vesicles in the optimized formulation. Overall, formulation F2 exhibited superior physicochemical stability, high entrapment efficiency, and favorable vesicle characteristics, establishing it as the most effective carrier system for topical antifungal therapy. These findings provide a foundation for further in vitro, ex vivo, and clinical investigations.