The aim of this research is undertaken to formulate thermo sensitive drug delivery system of Clonidine Hydrochloride for glaucoma therapy in in-situ form to overcome the problem of rapid precorneal elimination, poor bioavailability and nasolachrymal drainage exhibited by conventional ocular formulation. Thermoreversible ophthalmic drop were prepared using cold method by mixing Poloxamer 407 as thermosensitive polymer, HPMC K15M as viscosity enhancing agent, antiglaucoma drug Clonidine Hydrochloride and Benzalkonium Chloride as preservative. The in-situ gels were evaluated for gelation temperature, drug content, bioadhesive strength, viscosity and in-vitro release. A 22 factorial design employed for optimization of Clonidine Hydrochloride gels with Poloxamer 407 amount (% X1) and polymer (% HPMC K15M, X2) as the prime selected independent variable, which were varied at 2 different levels (low and high). The effect of formulation variables on response variable were statistically evaluated by using trial version of Design Expert® SOFTWAR (version 126.96.36.199). The result revealed that as the increase of viscosifying polymer HPMC K 15 M concentration, decrease in gelation temperature. pH of all formulation were found to be within the range of 6 to 6.8. The increase in level of HPMC K15M, the mucoadhesive strength increases. This developed formulation had optimum viscosity. The optimized formulation shows the controlled drug release. This study further demonstrates that administration of Clonidine Hydrochloride in the form of ocular gel is a pleasant, safe and effective.