Antibody specifically binding to GLP-1R and fusion protein thereof with GLP-1

• US 10,253,103 B2
• Filed: 08/13/2018
• Issued: 04/09/2019
• Est. Priority Date: 08/13/2013
• Status: Active Grant

• Current Assignee: Gmax Biopharm LLC
• Original Assignee: Gmax Biopharm LLC
• Inventors: Zhang, Cheng, Jing, Shuqian, Zhang, Hua, Wang, Xiaofeng, Yao, Chenjiang
• Primary Examiner(s): Li, Ruixiang
• Application Number:
  US16/102,552
• Publication Number:
  US 20180346585A1
• Time in Patent Office: 239 Days
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/605 Glucagons
• CPC Class Codes: C07K 16/2869 against hormone receptors for antibodies against neuromediator receptors C07K16/286
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/75 Agonist effect on antigen
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/31 fusions, other than Fc, for prolonged plasma life, e.g. albumin

Abstract

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.

Claims

1. A method of treating non-insulin-dependent diabetes in a subject, comprising administering to the subject a therapeutically effective amount of a GLP-1 fusion protein comprising a GLP-1 and an antibody specifically binding to GLP-1R, wherein the antibody comprises:

heavy chain CDR1 amino acid sequence;

SEQ ID NO;

12, heavy chain CDR2 amino acid sequence;

SEQ ID NO;

19, heavy chain CDR3 amino acid sequence;

SEQ ID NO;

27, light chain CDR1 amino acid sequence;

SEQ ID NO;

37, light chain CDR2 amino acid sequence;

SEQ ID NO;

45, and light chain CDR3 amino acid sequence;

SEQ ID NO;

53.

2. The method of claim 1, wherein the antibody comprises light chain variable region sequence:

SEQ ID NO;

101 or SEQ ID NO;

105; and

heavy chain variable region sequence;

SEQ ID NO;

75 or SEQ ID NO;

79.

3. The method of claim 1, wherein the antibody comprises a light chain variable region sequence encoded by polynucleotide sequence:

SEQ ID NO;

100 or SEQ ID NO;

104; and

a heavy chain variable region sequence encoded by polynucleotide sequence;

SEQ ID NO;

74 or SEQ ID NO;

78.

4. The method of claim 1, wherein the antibody comprises light chain variable region sequence:

SEQ ID NO;

101; and

heavy chain variable region sequence;

SEQ ID NO;

75.

5. The method of claim 1, wherein the antibody comprises a light chain variable region sequence encoded by polynucleotide sequence:

SEQ ID NO;

100; and

a heavy chain variable region sequence encoded by polynucleotide sequence;

SEQ ID NO;

74.

6. The method 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.

7. The method of claim 1, wherein the antibody comprises a light chain variable region sequence encoded by polynucleotide sequence:

SEQ ID NO;

104; and

a heavy chain variable region sequence encoded by polynucleotide sequence;

SEQ ID NO;

78.

8. The method of claim 1, 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.

9. The method of claim 1, wherein the antibody is a humanized antibody.

10. The method 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.

11. The method of claim 1, wherein the GLP-1 comprises amino acid sequence SEQ ID NO:

3, SEQ ID NO;

4, or SEQ ID NO;

5.

12. The method of claim 1, wherein the GLP-1 comprises amino acid sequence SEQ ID NO:

5.

13. The method of claim 1, wherein the GLP-1 is fused with a light chain of the antibody via a peptide linker.

14. The method of claim 13, wherein the peptide linker comprises amino acid sequence SEQ ID NO:

110, SEQ ID NO;

111, or SEQ ID NO;

112.

15. The method of claim 1, 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-R1r-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;

R1r 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.

16. A method of treating obesity in a subject, comprising administering to the subject a therapeutically effective amount of a GLP-1 fusion protein comprising a GLP-1 and an antibody specifically binding to GLP-1R, wherein the antibody comprises:

heavy chain CDR1 amino acid sequence;

SEQ ID NO;

12, heavy chain CDR2 amino acid sequence;

SEQ ID NO;

19, heavy chain CDR3 amino acid sequence;

SEQ ID NO;

27, light chain CDR1 amino acid sequence;

SEQ ID NO;

37, light chain CDR2 amino acid sequence;

SEQ ID NO;

45, and light chain CDR3 amino acid sequence;

SEQ ID NO;

53.

18. The method of claim 16, wherein the antibody comprises light chain variable region sequence:

SEQ ID NO;

101 or SEQ ID NO;

105; and

heavy chain variable region sequence;

SEQ ID NO;

75 or SEQ ID NO;

79.

19. The method of claim 16, wherein the antibody comprises a light chain variable region sequence encoded by polynucleotide sequence:

SEQ ID NO;

100 or SEQ ID NO;

104; and

a heavy chain variable region sequence encoded by polynucleotide sequence;

SEQ ID NO;

74 or SEQ ID NO;

78.

20. The method of claim 16, wherein the antibody comprises light chain variable region sequence:

SEQ ID NO;

101; and

heavy chain variable region sequence;

SEQ ID NO;

75.

21. The method of claim 16, wherein the antibody comprises a light chain variable region sequence encoded by polynucleotide sequence:

SEQ ID NO;

100; and

a heavy chain variable region sequence encoded by polynucleotide sequence;

SEQ ID NO;

74.

22. The method of claim 16, wherein the antibody comprises light chain variable region sequence:

SEQ ID NO;

105; and

heavy chain variable region sequence;

SEQ ID NO;

79.

23. The method of claim 16, wherein the antibody comprises a light chain variable region sequence encoded by polynucleotide sequence:

SEQ ID NO;

104; and

a heavy chain variable region sequence encoded by polynucleotide sequence;

SEQ ID NO;

78.

24. The method of claim 16, 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.

25. The method of claim 16, wherein the antibody is a humanized antibody.

26. The method of claim 16, 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.

27. The method of claim 16, wherein the GLP-1 comprises amino acid sequence SEQ ID NO:

3, SEQ ID NO;

4, or SEQ ID NO;

5.

28. The method of claim 16, wherein the GLP-1 comprises amino acid sequence SEQ ID NO:

5.

29. The method of claim 16, wherein the GLP-1 is fused with a light chain of the antibody via a peptide linker.

30. The method of claim 29, wherein the peptide linker comprises amino acid sequence SEQ ID NO:

110, SEQ ID NO;

111, or SEQ ID NO;

112.

31. The method of claim 16, 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-R1r-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;

R1r 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.

17. A method of treating overweight in a subject, comprising administering to the subject a therapeutically effective amount of a GLP-1 fusion protein comprising a GLP-1 and an antibody specifically binding to GLP-1R, wherein the antibody comprises:

heavy chain CDR1 amino acid sequence;

SEQ ID NO;

12, heavy chain CDR2 amino acid sequence;

SEQ ID NO;

19, heavy chain CDR3 amino acid sequence;

SEQ ID NO;

27, light chain CDR1 amino acid sequence;

SEQ ID NO;

37, light chain CDR2 amino acid sequence;

SEQ ID NO;

45, and light chain CDR3 amino acid sequence;

SEQ ID NO;

53.