The present research work was to formulate and evaluate Valacyclovir by using Box-Behnken Design, the current study intends to create and assess valacyclovir niosomes. Method: By employing the non-ionic surfactant Brij 72, the charge inducer diacetyl phosphate and cholesterol, stable Valacyclovir-loaded niosomes can be created. To study and optimize the primary effects, interaction effects and quadratic effects of the formulation ingredients on the performance of the niosomes, a total of 17 (VF1-VF17) formulations with the above concentrations was prepared using a three-factor, three-level Box-Behnken design. Results: Preformulation studies and drug excipients compatibility studies was done initially and results directed the further course of formulation. Various evaluation tests such as Zeta potential, Drug Content, Polydispersity Index, Entrapment Efficiency and Cumulative percent of drug released were performed to optimize the formulation. Most of the vesicles are spherical in shape, the size range of the vesicles fall in the narrow size range. A high % of valacyclovir can be encapsulated in the vesicles (72-86%) prepared by thin film hydration technique. Concentration of non-ionic surfactant and charge inducer might influence the drug release pattern of all formulation. In-vitro release of valacyclovir from niosomes was very slow and maintained prolong release, when compared to the release from pure valacyclovir solution. The Cumulative percent drug released of all prepared formulation shown (VF1-VF17) between 78.43% - 98.91%. Among the all formulations VF13 had shown 98.91% of drug release at 24 hrs. Conclusion: The niosomal formulation was stable, according to drug release experiments. Based upon all the evaluation tests, the formulation VF13 was considered as optimized. In all formulations, drug release was observed in all formulations indicating zero order release pattern.