Objective To prepare poloxamer 407-based in situ gel and to discover the relationship between temperature and concentration of polaxamer 407.Methods The pilocarpine-loaded in situ gel was prepared based on poloxamer 407,and the best concentration of poloxamer 407 was chosen according to the phase transfer temperature.Results Phase transfer temperature of gel with 18~20% polaxamer 407 was below human temperature,and the pH≈7.The in vitro release experiment showed that the drug release velocity of 18%,19%,20% in situ gels were 0.462,0.393,and 0.294 mg·min-1.Conclusion Dissolution of 18~20% poloxamer solution was suitable for ophthalmic drug delivery as the thermosensitive gel.