Sustainable, cost-efficient manufacturing of therapeutic peptides using chemo-enzymatic peptide synthesis (CEPS)
Using a large scale biocatalytic manufacture of the antidiabetic exenatide as an example, we report that chemo-enzymatic peptide synthesis (CEPS) constitutes an efficient method for sustainable manufacturing of therapeutic peptides. […]
Using a large scale biocatalytic manufacture of the antidiabetic exenatide as an example, we report that chemo-enzymatic peptide synthesis (CEPS) constitutes an efficient method for sustainable manufacturing of therapeutic peptides. The H-22-39-NH2 and H-1-21-O-Cam-L-NH2 exenatide fragments requisite for the enzymatic ligations were synthesized by solid-phase peptide synthesis (SPPS) methods and in ligations of these fragments catalyzed by omniligase-1, a broad specificity ligase engineered from subtilisin BPN’, we found that at pH 9.1/37 °C a substantial formation of deamidation impurities occurs. Consequently, a more robust ligation protocol (pH 7.5, rt, 10% MeCN as co-solvent) was developed and used in efficient scale-up ligations of the two fragments both as crude peptides as well as in their purified forms. Further, the aromatic 4-hydroxymethylbenzoic acid (HMBA) was evaluated as a more robust alternative for the carboxamidomethyl (O-Cam linker), which exhibited limited stability during some steps of the elevated temperature (ET) SPPS of the H-1-21-O-Cam-L-NH2 fragment.
J. Pawlas, T. Nuijens, J. Persson, T. Svensson, M. Schmidt, A. Toplak, M. Nilsson and J. H. Rasmussen, Green Chem., 2019, 21, 6451,
DOI: 10.1039/c9gc03600h