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Diabetes mellitus is an incurable metabolic disease characterized by a change in blood glucose level. Luteolin (LT) is a bioactive flavonoid and reported for potential antidiabetic activity. LT loaded chitosan (CH)—polylactic co-glycolic acid (PLGA) nanoparticles (NPs) were prepared by emulsification—evaporation process and further optimized by Box-Behnken design using CH (A), PLGA (B), and polyvinyl alcohol (C) as formulation variables and their effect evaluated on particle size (Y₁), drug loading (Y₂), and entrapment efficiency (Y₃). CH-PLGA-LT-NPs showed a particle size of 273.43 ± 5.24 nm, DL of 15.25 ± 2.43%, and EE of 65.28 ± 1.76%. The zeta potential was found to be positive with spherical surface morphology. CH-PLGA-LT-NPs exhibited sustained drug release (68.23 ± 6.17%) than LT-dispersion (29.66 ± 4.2%). The permeation study and pharmacokinetic study results revealed 3.19 fold …