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Sertaconazole nitrate is a novel broad spectrum antifungal drug with poor aqueous solubility which acted as a barrier against its clinical efficacy. Therefore, the aim of this work was to develop Sertaconazole nitrate loaded transfersomes using thin-film hydration method, based on a 23 factorial design. Different formulation variables were examined, namely, ratio of phospholipid to drug, ratio of phospholipid to Brij®L4 and method of controlling size of vesicles, and their effects on the entrapment efficiency (EE%), particle size (PS), polydispersity index (PDI), and zeta potential (ZP) were evaluated. Based on the desirability factor (0.898), the optimized formulation (F5) was selected by Design expert software for further investigations. The optimized formulation was imaged by TEM which revealed nanosizedunilamellar vesicles. Moreover, F5 improved the drug permeation and retention in the ex vivo permeation studies when compared to the STZ-suspension. Overall, the obtained results suggest that the fabricated formulation could be a promising vehicle for the dermal delivery of Sertaconazole nitrate.