Synthesis of end-labeled multivalent ligands for exploring cell-surface-receptor-ligand interactions.

BACKGROUND Ring-opening metathesis polymerization (ROMP) is a powerful synthetic method for generating unique materials. The functional group tolerance of ruthenium ROMP initiators allows the synthesis of a wide range of biologically active polymers. We generated multivalent ligands that inhibit cell surface L-selectin, a protein that mediates lymphocyte homing and leukocyte recruitment in inflammation. We hypothesized that these ligands function through specific, multivalent binding to L-selection. To examine this and to develop a general method for synthesizing multivalent materials with end-labels, we investigated functionalized enol ethers as capping agents in ruthenium-initiated ROMP. RESULTS We synthesized a bifunctional molecule that introduces a unique end group by terminating ruthenium-initiated ROMP reactions. This agent contains an enol ether at one end and a masked carboxylic acid at the other. We conjugated a fluorescein derivative to an end-capped neoglycopolymer that had previously been shown to inhibit L-selection function. We used fluorescence microscopy to visualize neoglycopolymer binding to cells displaying L-selectin. Our results suggest that the neoglycopolymers bind specifically to cell surface L-selectin through multivalent interactions. CONCLUSIONS Ruthenium-initiated ROMP can be used to generate biologically active, multivalent ligands terminated with a latent functional group. The functionalized polymers can be labeled with a variety of molecular tags, including fluorescent molecules, biotin, lipids or antibodies. The ability to conjugate reporter groups to ROMP polymers using this strategy has broad applications in the material and biological sciences.