Hydrophobicity as a tool for programming sequential mesophase transitions of enzyme-responsive polymeric amphiphiles

The ability of polymeric assemblies to undergo programmable cascades of mesophase transitions is prevalent in many systems in nature, where structural and functional features are tightly bound to maximize activity. In this study, we have examined the ability to program the mesophase transition rates of co-assembled enzyme-responsive polymeric micelles, through fine adjustments of the hydrophobicity of their amphiphilic components. We have utilized the different reactivities of di- and tri-block amphiphiles toward enzymatic degradation as a tool for programming formulations to undergo sequential enzymatically induced transitions from micelles to hydrogels and finally to dissolved polymers. By varying the aliphatic end-groups of PEG-dendron di-block and tri-block amphiphiles, we could demonstrate the remarkable impact of minor modifications to the di-block amphiphiles' structure and hydrophobicity on the transition rates between the different mesophases, ranging from a few hours to a week. Additionally, the study reveals how altering the relative hydrophobicity of its amphiphilic components influences the formulation ratio and enzymatic selectivity, as well as the stability and degradation rate of the resulting hydrogels. The findings underscore the importance of molecular architecture and hydrophobicity as key parameters in the design of programmable enzyme-responsive polymeric assemblies, offering insights into the ability to precisely control multi-step mesophase transitions for tailored functionality.