Methylphenidate, a CNS stimulant drug, has poor oral bioavailability due to extensive first-pass effect. In this study, Methylphenidate loaded transfersomes based in situ gel (MPH-TFs Gel) was prepared for brain targeting through intranasal (IN) drug delivery. Methylphenidate transfersomes (MPH-TFs) were prepared using the thin film hydration method. Box-Behnken Design expert software was utilized for the optimization by assessing the effect of independent variables phospholipon 90-G, Tween-80, and Methylphenidate (MPH) on particle size (PS), zeta potential (ZP), Polydispersity index (PDI), and Entrapment Efficiency (EE). The observed mean particle size of the optimized MPH-TFs was found to be 139.7 ± 3.75 nm, mono dispersion with PDI (0.208 ± 0.002), negatively charged (−27.9 ± 0.287 mV) and had an excellent entrapment efficiency (86 ± 3 %). Fourier-Transform Infrared (FTIR) spectra have depicted the compatibility of the components. X-r Diffraction analysis showed the conversion of crystalline structure methylphenidate to amorphous. The optimized MPH-TFs were then subsequently incorporated into an already optimized MPH-TFs Gel. MPH-TFs Gel was then characterized for its physicochemical properties, in vitro and in vivo studies. In vitro release and ex vivo permeation analysis showed sustained release and improved permeation of the MPH-TFs Gel, respectively. Additionally, histopathological studies of nasal membrane affirmed the safety of MPH-TFs Gel after intranasal application. In vivo pharmacokinetic study demonstrated improved brain bioavailability of MPH-TFs Gel compared to orally administered Methylphenidate solution (MPH-Sol).