Ribosomal Incorporation of N-Acetyl-3,5-Bis(Chloromethyl)benzylthio-L-Alanine for Developing an mRNA-Displayed Bicyclic Peptide Library Cyclized via 1,3,5-Tris(methyl)benzene

Abstract

We report a novel method for constructing an mRNA-displayed bicyclic peptide library cyclized through 1,3,5-tris(methyl)benzene by ribosomally incorporating N-acetyl-3,5-bis(chloromethyl)benzylthio-L-alanine, enabling the selection of bicyclic peptides against specific targets. Using this method, we successfully identified bicyclic peptides that bind to human Trop2 and VEGF165, providing a new strategy for selecting bicyclic peptides cyclized by trimethylbenzene.