The eye possesses numerous physiological barriers that significantly limit the absorption of administered drugs, as only <5 % get through. These barriers restrict drug bioavailability, often necessitating multiple dosing, which can lead to poor patient compliance. The study goal is to efficiently incorporate Voriconazole (VRC), a widespectrum antifungal candidate with a short half-life, into stable, biocompatible, non-irritating bile salt nanoparticles (Bilosomes, BLOs) to enhance its ocular delivery. Bilosomes were prepared by the ethanol injection method. Experimental (33 ) Box–Behnken Design (BBD) was employed; the amount of bile salt, cholesterol, and span-60 were chosen as independent variables, while entrapment efficiency (EE%), cumulative VRC release (Q8h), particle size (PS), zeta potential (ZP), and polydispersity index (PDI) were the dependent variables. The formula of optimization showed EE% (59.04 ± 1.88 %), cumulative release after 8 h (81.35 ± 0.69 %), PS (222.53 ± 2.04 nm), ZP (− 72.53 ± 1.75 mV), and PDI (0.225 ± 0.03). It was chemically characterized using DSC and FTIR. Its spherical arrangement was confirmed using TEM. For ophthalmic application, the optimized formula was included in ocular inserts to extend ocular residence time while minimizing dosing frequency. The optimal VRC-loaded bilosomal ocular insert met the acceptable physicochemical criteria, was sterilized using UV radiation for conducting microbiological assay, in-vivo study, and pharmacokinetic analysis. For safety evaluation, an eye irritation study and histopathological examination were conducted. The results confirmed its effectiveness as evidenced by a significantly larger inhibition zone and 8.39-fold enhancement in VRC bioavailability compared to its suspension. These findings support the potential of bilosomes-loaded ocular insert as an effective ocular delivery of VRC.