Improve Stability of Bioactive Peptides by Enzymatic Modular Synthesis of Peptides with O-Linked Sialyl Lewis x

Abstract

Glycosylation is an effective solution for peptide drug modification to overcome limitations such as instability under physiological conditions and lack of receptor selectivity. Disclosed herein is a facile enzymatic modular assembly strategy for the (semi)­preparative-scale synthesis of active glycopeptides. Sialyl Lewis x (sLex) oligosaccharides with E-selectin-targeting activity can be conjugated with peptides through multistep enzymatic reactions. The antiangiogenic peptide ES2 and anticancer peptide citropin 1.1 were successfully modified without loss of their biological activities. The half-lives of the glycopeptides were approximately 64-fold (ES2-sLex) and 28-fold (citropin-sLex) higher than that of the unmodified peptides in human serum.

Advances in biosynthesis of peptide drugs: Technology and industrialization

“…[66] Sialyl Lewis x (sLex) tetrasaccharide is an oligosaccharide structural unit at the surface of tumor cells and is able to bind to E-selectin on endothelial cells, so sLex-modified peptide drugs can be delivered to endothelial cells or nearby tumor cells. [67,68] Zheng et al [69] established an enzymatic modular assembly strategy to achieve sLex modification of peptide drugs, and synthesized gly-…”

“…[ 67,68 ] Zheng et al. [ 69 ] established an enzymatic modular assembly strategy to achieve sLex modification of peptide drugs, and synthesized glycopeptides in preparative or semi‐preparative scale. The glycosylation strategy links the shortest peptide sequence recognized by the O‐glycosyltransferase ncOGT (VPVSRA) to sLex by a multistep enzyme modularity reaction at the N‐terminal, C‐terminal or middle of the two active peptides to be modified.…”

Spore-Encapsulating Glycosyltransferase Catalysis Tandem Reactions: Facile Chemoenzymatic Synthesis of Complex Human Glycans

“…Although Sia-Gal moieties containing glycans have been demonstrated to play important roles in living cells, available methods to completely understand their precise functions and explore their medicinal application remain insufficient due to the lack of structurally defined substrates. , Hence, it is of great importance to develop methodologies to access these galactosylated and sialyl-galactosylated glycans in their pure forms. Chemical methods to synthesize Sia-Gal capped glycans are generally time-consuming and suffer from low yield given the presence of sialic acids, for which stereoselective introductions are challenging. , Recently, the development of enzymatic glycosylation reactions, which typically provide sufficient stereoselectivity, has overcome the above issue in synthesizing complex glycan structures. − Moreover, mammalian and microbial glycosyltransferases, such as α2,3-sialyltransferase 1 from Pasteurella multocida ( Pm 2,3ST1) and α2,6-sialyltransferase from Photobacterium damselae ( Pd 2,6ST), have been identified and readily expressed in Escherichia coli (E. coli), allowing the installation of corresponding glycosidic linkages. − Nevertheless, few methods have been reported to facilitate the preparation of these enzymes and purification of products from enzymatic reaction systems.…”