Aim:The study aimed to develop and optimize a thermosensitive in-situ gel of clarithromycin hydrochloride for ocular drug delivery, enhancing bioavailability and sustained drug release. Background:Conventional eye drops suffer from rapid drug elimination due to tear turnover and blinking, leading to poor ocular retention and low bioavailability. Thermosensitive in-situ gels address this issue by transforming from sol to gel at ocular temperature, prolonging drug residence time and therapeutic efficacy. Materials and Methods:A Box-Behnken design was used to optimize the formulation by varying Poloxamer 407, Poloxamer 188, and Carbopol 934P concentrations. The formulations were evaluated for cgelation temperature, viscosity, in vitro drug release, mean release time (MRT), antimicrobial activity, clarity, gel strength, gelling capacity, isotonicity studies, UV, FTIR, DSC, Dissolutionandstability study. The optimized formulation was selected based on ideal gelation behavior and sustained release characteristics. Results:The optimized formulation exhibited a gelation temperature of 34.0°C, ensuring immediate gel formation upon administration. The in vitro drug release study demonstrated a sustained release of 90.5% over 24 hours with an MRT of 7.2 hours. The formulation showed superior antimicrobial activity against Staphylococcus aureus and Pseudomonas aeruginosa. Stability studies confirmed no significant changes in pH, viscosity, and drug content over three months. Conclusion:The optimized thermosensitive in-situ gel of clarithromycin hydrochloride offers enhanced ocular retention, sustained drug release, and improved patient compliance, making it a promising alternative to conventional eye drops.