<p>Prednisolone acetate (PA) is a corticosteroid which has been known for potent anti-inflammatory and immunosuppressive properties. For optimizing the potential benefits of PA and dealing with challenges associated with its formulation, the novel drug delivery carrier-based topical formulation can be employed to target the drug at the site of inflammation. The objective of the research is to develop and optimize the PA-loaded NLC system and its incorporation into in situ gel to improve ocular retention, and extend drug release. In this study, Prednisolone acetate was successfully incorporated into nanostructured lipid carriers (NLC) by employing high-speed homogenization for ocular delivery. The optimized NLC formulation exhibited high encapsulation efficiency, smaller particle size, and acceptable polydispersity index. For improving ocular retention, NLC was transformed into an in situ gel developed using sodium alginate and hydroxypropyl methylcellulose. The formulation showed super gelation characteristics with high viscosity and strong gel network which is vital for its retention on the ocular region. Further, the adhesiveness value of 29.98&#xa0;N/mm indicated the exceptional mucoadhesive properties. The in vitro release studies indicated that NLC-based in situ gel sustained the drug release for prolonged period showing 61.58 and 98.2% of drug release after 9 and 24&#xa0;h, respectively. The release kinetic studies showed that optimized formulation followed Hixson-Crowell model, suggesting a surface area-dependent drug release profile. These findings suggest that NLC-incorporated in in situ gel systems act as a promising alternative drug delivery platform for ocular administration by enhancing precorneal residence time and facilitating sustained drug release at the target site.</p>