Unravelling Structure-function Interactions Between Fluorinated Heparan Sulfate Mimetics and Signaling Proteins

Abstract

Fluorinated carbohydrates are emerging scaffolds in glycobiology, enabling the elucidation of the individual hydroxyl groups roles of a carbohydrate in protein binding and drug discovery. Herein, we report the divergent strategy to synthesize seven heparan sulfate (HS) mimetics featuring a fluorine atom at the C3 position of the glucuronic acid residue, with the objective of modulating structure-function relationships. The sensitivity of fluorine signals to sulfation patterns was confirmed via¹⁹F-NMR, while ³J_HH coupling and NOE data demonstrated that the glucuronic acid residue retained its ⁴C_1​ conformation. Glycan microarray analysis and SPR binding studies revealed that a single hydroxyl-to-fluorine substitution in HS mimetics retains the binding of N-acetylated HS sequences for several growth factors and chemokines. Remarkably, GlcNAc6S-GlcA(3F) and GlcNS6S3S-GlcA(3F) exhibited binding properties comparable to those of highly N-sulfated native HS ligands. These findings provide valuable insights for development of novel therapeutic agents targeting morphogens and cell signalling pathways.