Glycosylated Form of Antigenic GLP-1 Analogue
Disclosed is a glucagon-like peptide-1 (GLP-1) analogue which is obtained by ameliorating a highly antigenic GLP-1 analogue so that the GLP-1 analogue has reduced antigenicity without being lowered in the blood glucose suppressing activity. Specifically disclosed is a glycosylated form of an antigenic GLP-1 analogue, which has GLP-1 activity and is obtained by substituting at least one amino acid of an antigenic GLP-1 analogue with a glycosylated amino acid.
1 . An oligosaccharide chain added form of antigenic glucagon-like peptide-1 (GLP-1) analogue having GLP-1 activity, wherein at least one amino acid of the antigenic GLP-1 analogue is substituted with an oligosaccharide chain added amino acid.
2 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 1 , wherein said oligosaccharide chain added amino acid is an oligosaccharide chain added Asn or an oligosaccharide chain added Cys.
3 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 1 , wherein the oligosaccharide chain and the amino acid are bound via a linker in said oligosaccharide chain added amino acid.
4 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 1 , wherein said oligosaccharide chain is an oligosaccharide chain consisting of 4 or more sugars.
5 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 1 , wherein said oligosaccharide chain is a double-stranded complex oligosaccharide chain.
6 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 5 , wherein said oligosaccharide chain is an oligosaccharide chain selected from disialooligosaccharide chain, monosialooligosaccharide chain, asialooligosaccharide chain, diGlucNAc oligosaccharide chain, and dimannose oligosaccharide chain.
7 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 5 , wherein said oligosaccharide chain is an oligosaccharide chain represented by:
wherein R 1 and R 2 are the same or different, and indicate:
and Ac indicates an acetyl group.
8 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 1 , wherein said antigenic GLP-1 analogue is exendin-4.
9 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 8 , wherein exendin-4 has an amino acid sequence of SEQ ID NO:2 and the site substituted with an oligosaccharide chain added amino acid is position 30 in the amino acid sequence set forth in SEQ ID NO: 2.
10 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 8 ,
wherein exendin-4 has an amino acid sequence of SEQ ID NO:2 and in the amino acid sequence set forth in SEQ ID NO: 2, Gly at position 30 is substituted with oligosaccharide chain added Cys
wherein said oligosaccharide chain is the disialooligosaccharide chain shown by the following formula:
and
wherein said oligosaccharide chain and said Cys are bound via a linker in said oligosaccharide chain added Cys.
11 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 1 , wherein said antigenic GLP-1 analogue is liraglutide.
12 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 11 , wherein said site substituted with an oligosaccharide chain added amino acid is position 30 in the amino acid sequence set forth in SEQ ID NO: 3.
13 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 11 ,
wherein in the peptide consisting of the amino acid sequence set forth in SEQ ID NO: 3, Arg at position 30 is substituted to oligosaccharide chain added Cys,
wherein said oligosaccharide chain is the disialooligosaccharide chain represented by the following formula:
or the asialooligosaccharide chain represented by the following formula:
and
wherein said oligosaccharide chain and said Cys are bound via a linker in said oligosaccharide chain added Cys.
14 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 1 , wherein said oligosaccharide chain is substantially homogeneous.
15 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 1 , wherein said oligosaccharide chain is 99% or more homogeneous.
16 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 1 , wherein the antigenicity is half or less relative to an antigenic GLP-1 analogue without any oligosaccharide chains added thereto.
17 . The oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 1 , wherein the antigenicity is reduced compared to the antigenic GLP-1 analogue without any oligosaccharide chains added thereto, and GLP-1 activity is elevated compared to natural form GLP-1.
18 . A pharmaceutical composition comprising the oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 1 as an active ingredient.
19 . The pharmaceutical composition according to claim 18 for therapy or prevention of a GLP-1-related disease.
20 . The pharmaceutical composition according to claim 19 , wherein said GLP-1-related disease is diabetes.
21 . A method of treating or preventing a GLP-1-related disease in a subject, comprising administering to said subject an effective amount of the oligosaccharide chain added form of antigenic GLP-1 analogue according to claim 1 .