The present study was planned to investigate the effect of polymer composites HPMC K15 M and poloxamer 407 on developed dorzolamide hydrochloride in situ gel. Formulations were prepared based on 32 factorial design with concentrations of Poloxamer 407 and HPMC K15 M as independent variables. Gelation temperature, viscosity study at 37 °C and % cumulative drug release upto 8 h were considered as dependent variables. Polymer composites showed linear model with gelation temperature and quadratic model with viscosity study at 37 °C and % cumulative drug release up to 8 h. Optimized formulation successfully sustained release of drug upto 5 h in ex vivo goat corneal permeability study. Comparative in vivo study in normotensive rabbits showed that optimized formulation sustained therapeutic effect upto 8 h with 31.22 ± 3.65% reduction in intraocular pressure and marketed formulation showed immediate release effect with 18.22 ± 4.42% reduction in intraocular pressure upto 2-3 h.