Antibody specifically binding to GLP-1 R and fusion protein thereof with GLP-1
Disclosed in the present invention is an antibody specifically binding to GLP-1R and a fusion protein thereof with GLP-1. The fusion proteins can effectively bind to a human GLP-1R receptor and activate a receptor signaling pathway, thus are useful for treating diabetes, excessive weight, obesity and related disorders thereof.
1. An antibody specifically binding to GLP-1R, comprising:
(a) light chain CDR1 amino acid sequence: SEQ ID NO: 37;
(b) light chain CDR2 amino acid sequence: SEQ ID NO: 45;
(c) light chain CDR3 amino acid sequence: SEQ ID NO: 53;
(d) heavy chain CDR1 amino acid sequence: SEQ ID NO: 12;
(e) heavy chain CDR2 amino acid sequence: SEQ ID NO: 19; and
(f) heavy chain CDR3 amino acid sequence: SEQ ID NO: 27.
2. The antibody of claim 1 , wherein the antibody comprises light chain variable region sequence: SEQ ID NO: 105 and heavy chain variable region sequence: SEQ ID NO: 79.
3. The antibody of claim 2 , wherein the antibody further comprises an amino acid sequence selected from:
(a) light chain constant region amino acid sequence: SEQ ID NO 106;
(b) light chain constant region amino acid sequence: SEQ ID NO 107;
(c) heavy chain constant region amino acid sequence: SEQ ID NO 108;
(d) heavy chain constant region amino acid sequence: SEQ ID NO 109;
(e) light chain constant region amino acid sequence: SEQ ID NO 106 and heavy chain constant region amino acid sequence: SEQ ID NO 108;
(f) light chain constant region amino acid sequence: SEQ ID NO 107 and heavy chain constant region amino acid sequence: SEQ ID NO 108;
(g) light chain constant region amino acid sequence: SEQ ID NO 106 and heavy chain constant region amino acid sequence: SEQ ID NO 109; and
(h) light chain constant region amino acid sequence: SEQ ID NO 107 and heavy chain constant region amino acid sequence: SEQ ID NO 109.
4. A GLP-1 fusion protein comprising GLP-1 and the antibody of claim 1 , wherein the GLP-1 comprises an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 127.
5. The GLP-1 fusion protein of claim 4 , wherein the GLP-1 is fused with the light chain and/or heavy chain of the antibody in the form of N′-R1-L-R2-C′, N′-R2-L-R1-C′ or N′-R2-R1 r -C′,
wherein:
L is a peptide linker, comprising a full-length, partial, or repeated amino acid sequence selected from SEQ ID NO: 110, SEQ ID NO: 111, and SEQ ID NO: 112;
R1 is an amino acid sequence of GLP-1;
R1 r is a reverse amino acid sequence of GLP-1;
R2 is an amino acid sequence of the light chain or heavy chain of the antibody;
C′ represents a carboxyl residual terminal of the GLP-1 fusion protein polypeptide; and
N′ represents an amino residual terminal of the GLP-1 fusion protein polypeptide.
6. A pharmaceutical composition comprising the GLP-1 fusion protein of claim 4 and a pharmaceutically acceptable carrier.
7. The GLP-1 fusion protein of claim 4 , wherein the GLP-1 comprises amino acid sequence SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5.
8. The GLP-1 fusion protein of claim 4 , wherein the GLP-1 comprises amino acid sequence SEQ ID NO: 5.
9. The antibody of claim 1 , wherein the antibody is a monoclonal antibody.
10. The antibody of claim 1 , wherein the antibody is a humanized antibody.
11. A GLP-1 fusion protein comprising the antibody of claim 1 and GLP-1, wherein the GLP-1 comprises amino acid sequence SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5.
12. The GLP-1 fusion protein of claim 11 , wherein the GLP-1 comprises amino acid sequence SEQ ID NO: 5.
13. The GLP-1 fusion protein of claim 11 , wherein the GLP-1 is fused with a light chain of the antibody via a peptide linker.
14. The GLP-1 fusion protein of claim 11 , wherein the GLP-1 is fused with a light chain of the antibody in the form of N′-R1-L-R2-C′,
wherein:
L is a peptide linker comprising amino acid sequence SEQ ID NO: 110, SEQ ID NO: 111, or SEQ ID NO: 112;
R1 is the GLP-1;
R2 is a light chain of the antibody;
C′ represents a carboxyl residual terminal of the GLP-1 fusion protein polypeptide; and
N′ represents an amino residual terminal of the GLP-1 fusion protein polypeptide.
15. The GLP-1 fusion protein of claim 11 , wherein the peptide linker comprises amino acid sequence SEQ ID NO: 110, SEQ ID NO: 111, or SEQ ID NO: 112.
16. A pharmaceutical composition comprising the GLP-1 fusion protein of claim 11 and a pharmaceutically acceptable carrier.
17. A pharmaceutical composition comprising the antibody of claim 1 and a pharmaceutically acceptable carrier.