Chemoenzymatic synthesis of glycosylated glucagon-like peptide 1: Effect of glycosylation on proteolytic resistance and in vivo blood glucose-lowering activity

Taichi Ueda, Kazuyoshi Tomita, Yoshihide Notsu, Takaomi Ito, Masataka Fumoto, Tomoaki Takakura, Hirofumi Nagatome, Akio Takimoto, Shin Ichi Mihara, Hiroko Togame, Keiko Kawamoto, Takanori Iwasaki, Kenji Asakura, Takeo Oshima, Kohji Hanasaki, Shin Ichiro Nishimura, Hirosato Kondo

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Glucagon-like peptide 1 (7-36) amide (GLP-1) has been attracting considerable attention as a therapeutic agent for the treatment of type 2 diabetes. In this study, we applied a glycoengineering strategy to GLP-1 to improve its proteolytic stability and in vivo blood glucose-lowering activity. Glycosylated analogues with A/-acetylglucosamine (GlcNAc), A/-acetyllactosamine (LacNAc), and α2,6-sialyl N-acetyl-lactosamine (sialyl LacNAc) were prepared by chemoenzymatic approaches. We assessed the receptor binding affinity and cAMP production activity in vitro, the proteolytic resistance against dipeptidyl peptidase-IV (DPP-IV) and neutral endopeptidase (NEP) 24.11, and the blood glucose-lowering activity in diabetic db/db mice. Addition of sialyl LacNAc to GLP-1 greatly improved stability against DPP-IV and NEP 24.11 as compared to the native type. Also, the sialyl LacNAc moiety extended the blood glucose-lowering activity in vivo. Kinetic analysis of the degradation reactions suggested that the sialic acid component played an important role in decreasing the affinity of peptide to DPP-IV. In addition, the stability of GLP-1 against both DPP-IV and NEP24.11 incrementally improved with an increase in the content of sialyl LacNAc in the peptide. The di- and triglycosylated analogues with sialyl LacNAc showed greatly prolonged blood glucose-lowering activity of up to 5 h after administration (100 nmol/kg), although native GLP-1 showed only a brief duration. This study is the first attempt to thoroughly examine the effect of glycosylation on proteolytic resistance by using synthetic glycopeptides having homogeneous glycoforms. This information should be useful for the design of glycosylated analogues of other bioactive peptides as desirable pharmaceuticals.

Original languageEnglish
Pages (from-to)6237-6245
Number of pages9
JournalJournal of the American Chemical Society
Volume131
Issue number17
DOIs
Publication statusPublished - May 6 2009
Externally publishedYes

Fingerprint

Glycosylation
Glucagon-Like Peptide 1
Dipeptidyl Peptidase 4
Peptides
Glucose
Blood Glucose
Blood
Neprilysin
Medical problems
Amides
Acetylglucosamine
Drug products
Glycopeptides
N-Acetylneuraminic Acid
Type 2 Diabetes Mellitus
Degradation
Kinetics
Acids
Peptide Hydrolases
Pharmaceutical Preparations

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Chemoenzymatic synthesis of glycosylated glucagon-like peptide 1 : Effect of glycosylation on proteolytic resistance and in vivo blood glucose-lowering activity. / Ueda, Taichi; Tomita, Kazuyoshi; Notsu, Yoshihide; Ito, Takaomi; Fumoto, Masataka; Takakura, Tomoaki; Nagatome, Hirofumi; Takimoto, Akio; Mihara, Shin Ichi; Togame, Hiroko; Kawamoto, Keiko; Iwasaki, Takanori; Asakura, Kenji; Oshima, Takeo; Hanasaki, Kohji; Nishimura, Shin Ichiro; Kondo, Hirosato.

In: Journal of the American Chemical Society, Vol. 131, No. 17, 06.05.2009, p. 6237-6245.

Research output: Contribution to journalArticle

Ueda, T, Tomita, K, Notsu, Y, Ito, T, Fumoto, M, Takakura, T, Nagatome, H, Takimoto, A, Mihara, SI, Togame, H, Kawamoto, K, Iwasaki, T, Asakura, K, Oshima, T, Hanasaki, K, Nishimura, SI & Kondo, H 2009, 'Chemoenzymatic synthesis of glycosylated glucagon-like peptide 1: Effect of glycosylation on proteolytic resistance and in vivo blood glucose-lowering activity', Journal of the American Chemical Society, vol. 131, no. 17, pp. 6237-6245. https://doi.org/10.1021/ja900261g
Ueda, Taichi ; Tomita, Kazuyoshi ; Notsu, Yoshihide ; Ito, Takaomi ; Fumoto, Masataka ; Takakura, Tomoaki ; Nagatome, Hirofumi ; Takimoto, Akio ; Mihara, Shin Ichi ; Togame, Hiroko ; Kawamoto, Keiko ; Iwasaki, Takanori ; Asakura, Kenji ; Oshima, Takeo ; Hanasaki, Kohji ; Nishimura, Shin Ichiro ; Kondo, Hirosato. / Chemoenzymatic synthesis of glycosylated glucagon-like peptide 1 : Effect of glycosylation on proteolytic resistance and in vivo blood glucose-lowering activity. In: Journal of the American Chemical Society. 2009 ; Vol. 131, No. 17. pp. 6237-6245.
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