Synthesis of Degradable Glycopolymers and Programming of Interfacial Hydrophobic Patches for Optimal TLR4-Mediated Immunity

Synthetic glycopolymers are potent immunomodulators, yet the role of programmable interfacial cues in engaging Toll-like receptors (TLRs) remains poorly defined. Here, we investigate the self-assembly behavior of degradable amphiphilic glycopolymers synthesized via radical ring-opening copolymerization. We demonstrate that dendritic cell (DC) maturation exhibits a pronounced nonlinear dependence on backbone hydrophobicity, with an MDO molar fraction between 30 and 40% identified as the “sweet spot” for maximizing DC maturation markers (CD80 + /CD86 +, MHC II, and CCR7). By integrating ANS fluorescence assays, multiscale DPD simulations, and molecular docking, we confirm that this dependence is primarily dictated by the formation of surface “hydrophobic patches.” Specifically, a surface hydrophobic patch content of ∼29% (exemplified by the PGM-4 formulation) provides the ideal interface for TLR4-associated signaling by optimizing the balance between hydrophobic anchoring and multivalent glycan recognition, as evidenced by inhibition studies with the TLR4-selective antagonist TAK-242. Furthermore, these nanoparticles maintain sufficient structural stability in the short term while exhibiting enzyme-responsive backbone degradation over a longer duration. This study establishes a quantitative framework for designing “patchy” glyco-nanoparticles, where the synergy between hydrophobic domains and glycan clusters dictates potent adjuvant activity.