Abstract: Amisulpride is practically insoluble in water and suffers from irregular and low bioavailability (48%). This could be due to low solubility and being a substrate for P-glycoprotein efflux. The aim of this study is to develop and statistically optimize Self-Emulsifying Drug Delivery System (SEDDS) formulation containing bio-enhancers and P-glycoprotein inhibitors components, for the improvement of dissolution and oral absorption of amisulpride; using Central Composite Rotatable Design (CCRD). Preliminary screening was carried out to determine amisulpride solubility in various oils and surfactants. Formulations were prepared using oil (Capryol-90®), two surfactants (Cremophor EL® and Labrasol®, “Smix”) and co-surfactant (Transcutol HP®). CCRD was applied for optimization. Oil percentage, Smix: co-surfactant ratio and Cremophor EL: Labrasol ratio in Smix, were selected as independent variables while mean droplet size, drug loading and light absorbance of diluted SEDDS as dependent variables. Second-order polynomial equations were fitted to data. Optimized formulation, containing 10% oil, 1.31 as Smix: co-surfactant ratio and 2 as ratio of Cremophor ®EL: Labrasol® in Smix was prepared according to software determined levels using desirability function and overlay plot. It provided drug loading of 50 mg mL-1 and released amisulpride completely within 15 min irrespective of type or pH of dissolution medium. Optimized SEDDS showed significant (p<0.01) increase in vivo bioavailability compared to drug suspension. CCRD could be considered as an efficient approach for the optimization of SEDDS. Also, the optimized SEDDS formulation demonstrated a great potential as a possible alternative to traditional oral formulations of amisulpride.