Enzymatic Glycosylation of Triazole-Linked GlcNAc/Glc-Peptides: Synthesis, Stability and Anti-HIV Activity of Triazole-Linked HIV-1 gp41 Glycopeptide C34 Analogues

• Published in: Chembiochem : a European journal of chemical biology 2009-05, Vol.10 (7), p.1234-1242
• Main Authors: Huang, Wei, Groothuys, Stan, Heredia, Alonso, Kuijpers, Brian H.M, Rutjes, Floris P.J.T, van Delft, Floris L, Wang, Lai-Xi
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Anti-HIV Agents - chemistry; Anti-HIV Agents - metabolism; Anti-HIV Agents - pharmacology; antiviral agents; Arthrobacter; Arthrobacter - enzymology; Biocatalysis; Carbohydrate Sequence; chemoenzymatic synthesis; Drug Stability; Glycopeptides - biosynthesis; Glycopeptides - chemistry; Glycopeptides - pharmacology; glycoproteins; Glycosylation; HIV Envelope Protein gp41 - chemistry; Human immunodeficiency virus; Human immunodeficiency virus 1; Inhibitory Concentration 50; Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase - metabolism; Molecular Sequence Data; protease resistance; Substrate Specificity; Triazoles - chemistry
• ISSN: 1439-4227; 1439-7633
• DOI: 10.1002/cbic.200800741

Long-lasting sweet proteins: The chemoenzymatic synthesis of a triazole (T)-linked glycosylated C34 fragment from HIV-1 gp41 is described. The glycopeptide shows high solubility, excellent fusion inhibition, and as shown in the graph, promising protease resistance.Endoglycosidase-catalyzed transglycosylation of triazole-linked glucose (Glc) and N-acetylglucosamine (GlcNAc)-containing dipeptides and polypeptides was achieved by using synthetic sugar oxazoline as the donor substrate. It was found that both N- and C-linked Glc/GlcNAc-containing triazole derivatives were effective substrates for endo-β-N-acetylglucosaminidase from Arthrobacter (Endo-A) for transglycosylation; this demonstrates a broad acceptor substrate specificity for Endo-A. This chemoenzymatic method was successfully used for the synthesis of a novel triazole-linked C34 glycopeptide derived from the HIV-1 envelope glycoprotein, gp41. We found that the synthetic C34 glycopeptide possesses potent anti-HIV activity with an IC₅₀ of 21 nM. The triazole-linked C34 glycopeptide demonstrated a much enhanced stability against protease- and glycoamidase-catalyzed digestion; this shows the protective effects of glycosylation and the stability of the triazole linkage. These favorable properties suggest that the triazole-linked C34 glycopeptide might be valuable for further development as an anti-HIV drug candidate.