The bioavailability of therapeutic agents from eye drops is usually limited due to corneal barrier functions and effective eye protective mechanisms. The current study aimed to enhance ocular bioavailability of dorzolamide hydrochloride, a potent antiglaucoma drug by preparing novel ophthalmic drug delivery system like soluble ophthalmic drug inserts (SODI). Various batches of dorzolamide hydrochloride Soluble Ocular Drug Insert were prepared using Polyvinyl Alcohol, Poloxamer 407 as polymer with Propylene Glycol as plasticizer by solvent casting method. The drug inserts were prepared by using 32 factorial designs (version 10). Solvent casting method was used to prepare drug inserts which were further evaluated for various parameters like drug interaction studies, physicochemical characteristics, in vitro release studies, transcorneal permeation study by using excised goat cornea. The optimized batch F2 showed 98.50 ± 0.05% drug release at the end of 6 h and has desired physicochemical properties. The gamma radiation sterilized batch F2 was subjected to sterility test as per Indian Pharmacopeia 2014 and accelerated stability studies as per International Conference on Harmonization stability testing of new drug substances and products guideline, ex vivo and Pharmacodynamic studies. The comparative pharmacokinetic result shows that Dorzolamide Hydrochloride SODI is more effective than the marketed formulation. The promising pharmacokinetic results and possibility of commercial applicability justifies the need of product in market.