This study is devoted to developing amphiphilic block polymers based on phenylborate ester, which can self-assemble to form nanoparticles, as a glucose-sensitive drug carrier. Poly(ethylene glycol)-block-poly[(2-phenylboronic esters-1,3-dioxane-5-ethyl) methylacrylate] (MPEG5000-block-PBDEMA) was fabricated with MPEG5000-Br as a macroinitiator via atom transfer radical polymerization (ATRP). Using the solvent evaporation method, these block polymers can disperse in aqueous milieu to self-assemble into micellar aggregates with a spherical core-shell structure. Zeta potential and fluorescence techniques analysis showed a good purification effect, high encapsulation efficiency, and loading capacity of fluorescein isothiocyanate (FITC)-insulin-loaded polymeric micelles under optimal conditions. The in vitro insulin release profiles revealed definite glucose-responsive behavior of the polymeric micelles at pH 7.4 and 37 °C, depending on the environmental glucose concentration and the chemical composition of the block polymers. Further, circular dichroism spectroscopy demonstrated that the overall tertiary structure of the released insulin was in great agreement with standard insulin. (1)H NMR results of the polymeric micelles during glucose-responsive process supposed one possible insulin release mechanism via the polymer polarity transition from amphiphilic to double hydrophilic. The analysis of L929 mouse fibroblast cells viability suggested that the polymeric micelles from MPEG5000-block-PPBDEMA had low cell toxicity. The block polymers containing phenylborate ester that responded to changes in the glucose concentration at neutral pH are being aimed for use in self-regulated insulin delivery.