Poster Presentation 9th Modern Solid Phase Peptide Synthesis & Its Applications Symposium 2023

The chemo-enzymatic synthesis of glycosylated semaglutide analogue (#107)

Feng Lin 1 , Chaitra Chandrashekar 1 , Sam van Dun 1 , Anna Koijen 2 , Leendert J. van den Bos 2 , Mohammed Akhter Hossain 1 , John D Wade 1
  1. Florey Neuroscience Institute, University of Melbourne, Melbourne, VIC 3010, Australia
  2. EnzyTag B.V., 6361 HK , Nuth, Netherlands

Semaglutide is one of Glucagon-like peptide-1 analogues, as a GLP-1 receptor agonist, used for the treatment of type 2 diabetes. The glycosylation of GLP-1 analogues, not only increased the half-life time of glycosylated exenatide in vitro serum [1], also reduced the fibrillation and aggregation of glycosylated glucagon analogue in an aqueous solution [2]. In order to enhance the yield of synthesis and improve the stability of semaglutide, the modified residue Lys20-(C18diacid-ꝩ-Glu-OEG-OEG) was replaced by Asn20-(disialyloligosacharide), we used our highly optimized Fmoc-SPPS methods to prepare each of Semaglutide (1-11)-Ocam-Leu amide and Semaglutide (12-31)-Asn20-(disialyloligosacharide), follow by enzymatic ligation of those fragments with Omniligase-1 [3], after RP-HPLC purification, glycosylated Semaglutide analogue was obtained in good overall yield. The bioassay result shown that its biological activity was similar with Semaglutide.

  1. Chaitra Chandrashekar, Yuji Nishiuchi, Barbara Fam White et al. (2023). Glycosylation improves the proteolytic stability of exenatide. Bioconjugate Chemistry 34, 6, 1014-1018.
  2. Mengjie Liu, Peishen Zhao, Md Hemayet Uddin et al. (2021). Chemical Synthesis and Characterization of a Nonfibrillating Glycoglucagon. Bioconjugate Chemistry 32, 10, 2148–2153.
  3. Jan pawlas, Timo Nuijens, Jonas Persson et al. (2019). Sustainable, cost-efficient manufacturing of therapeutic peptides using chemo-enzymatic peptide synthesis. Green Chemistry, 21, 6451-6467.