The formulation development of Glibenclamide (GBC) loaded nanostructured lipid carriers (NLCs) using Box-Behnken design is the focus of this study. GBC is a widely used antidiabetic drug, and the utilization of NLCs as carriers can enhance its bioavailability, stability, and targeted delivery. The Box-Behnken design, a statistical experimental design, is employed to optimize the formulation variables for achieving desirable NLC characteristics. In this study, NLCs were prepared using a solvent injection method. The independent variables considered for optimization were the concentration of solid lipid Glycerylmonostearate, concentration of liquid lipid (Caprol PGE-860, and the concentration of surfactant Pluronic F127. The dependent variables evaluated were entrapment efficiency and particle size. The Box-Behnken design allowed the investigation of 17 experimental runs, including both factorial and axial points. The obtained data were analyzed using response surface methodology to establish mathematical models correlating the independent and dependent variables. The models were validated, and the optimized formulation was determined using desirability function. The particle size of GBC-loaded NLCs for formulation F1 to F17 was found to be between 86.74 to 126.65nm respectively The Entrapment efficiency of GBC-loaded NLCs for formulation F1 to F17 was found to be between 60.74 to 73.32 percentages respectively. The Zeta Potential value of formulations OF1, OF2, and OF3 were found to be -35.65, -41.38 and -40.65 mV respectively.In comparison to all formulations, formulation OF3 showed sustain drug release from formulation up to 24 hrs. The formulation showed 13.25, 18.95, 22.23, 36.65, 48.85, 59.98, 69.89, 86.65 and 98.85 percentage after 0.5, 1, 2, 4, 6, 8, 10, 12 and 24 hrs. respectively. This study highlights the successful utilization of the BoxBehnken design in the formulation development of GBC-loaded NLCs. The optimized formulation exhibited desirable characteristics, indicating its potential as a delivery system for GBC in the treatment of diabetes. Further in vitro and in vivo studies are warranted to evaluate its pharmacokinetic and therapeutic efficacy