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

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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.

元の言語英語
ページ(範囲)6237-6245
ページ数9
ジャーナルJournal of the American Chemical Society
131
発行部数17
DOI
出版物ステータス出版済み - 5 6 2009
外部発表Yes

All Science Journal Classification (ASJC) codes

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

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    Ueda, T., Tomita, K., Notsu, Y., Ito, T., Fumoto, M., Takakura, T., Nagatome, H., Takimoto, A., Mihara, S. I., Togame, H., Kawamoto, K., Iwasaki, T., Asakura, K., Oshima, T., Hanasaki, K., Nishimura, S. I., & 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, 131(17), 6237-6245. https://doi.org/10.1021/ja900261g