Ribosomal Translation and Display Selection of Cyclic Glycopeptides through Genetic Reprogramming

A robust method for the ribosomal incorporation of glycosyl-amino acids through the flexizyme-mediated reprogramming of initiating AUG codons with glycosyl-dipeptides is described. This reprogramming strategy was applied to in vitro mRNA display for the de novo discovery of α-l-fucosylated cyclic peptide inhibitors of P-selectin with antiplatelet-monocyte aggregation activity, and β-d-N-acetylgalactosamine-containing cyclic glycopeptide agonists of the asialoglycoprotein receptor (ASGPR) with potent lysosome trafficking activity. This work provides a powerful platform for the de novo discovery of glycopeptide ligands and expands genetic reprogramming capabilities to large modifications previously intractable for ribosomal translation.