Effect of Brønsted Acids on the Activation of Mixed Anhydride/Mixed Carbonic Anhydride and C‐Terminal‐Free N‐Methylated Peptide Synthesis in a Micro‐Flow Reactor

Abstract

AbstractAmidations employing mixed (carbonic) anhydrides have long been favoured in peptide synthesis because of their cost‐effectiveness and less waste generation. Despite their long history, no study has compared the effects of additives on the activation of mixed anhydrides and carbonic anhydrides. In this study, we investigated the amidation of mixed (carbonic) anhydride in the presence of a base and/or Brønsted acids. The use of NMI⋅HCl significantly improved the conversion of the mixed carbonic anhydride, while expediting nucleophilic attacks on the desired carbonyl group. In contrast, in the case of mixed anhydrides, neither the conversion nor the desired nucleophilic attack improved significantly. We developed a C‐terminus‐free N‐methylated peptide synthesis method using mixed carbonic anhydrides in a micro‐flow reactor. Fourteen N‐alkylated peptides were synthesized in moderate to high yields (55–99 %) without severe racemization (<1 %). Additionally, a significant enhancement in the amidation between mixed carbonic anhydrides and bis‐TMS‐protected N‐methyl amino acids with the inclusion of NMI⋅HCl was observed for the first time. In addition, we observed unexpected C‐terminal epimerization of the C‐terminus‐free N‐methyl peptides.