ANTIBODY SPECIFICALLY BINDING TO GLP-1R AND FUSION PROTEIN THEREOF WITH GLP-1

• US 20160362498A1
• Filed: 08/01/2014
• Published: 12/15/2016
• Est. Priority Date: 08/13/2013
• Status: Active Grant

• Current Assignee: Gmax Biopharm LLC
• Original Assignee: Gmax Biopharm LLC
• Inventors: ZHANG, Cheng, JING, Shuqian, WANG, Xiaofeng, YAO, Chenjiang, ZHANG, Hua
• Granted Patent: US 10,059,773 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• 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. An antibody specifically binding to GLP-1R, comprising an amino acid sequence selected from:

(a) a light chain CDR3 sequence selected from;

light chain CDR3 sequences differing by no more than three amino acid additions, substitutions and/or deletions in total from one of L1-L13 light chain CDR3 sequences;

SEQ ID NO;

46 to SEQ ID NO;

53;

(b) a heavy chain CDR3 sequence selected from;

heavy chain CDR3 sequences differing by no more than four amino acid additions, substitutions and/or deletions in total from one of H1-H13 heavy chain CDR3 sequences;

SEQ ID NO;

20 to SEQ ID NO;

27; and

(c) a light chain CDR3 sequence from (a) and a heavy chain CDR3 sequence from (b).

2. The antibody of claim 1, further comprising one or more amino acid sequences selected from:

(a) a light chain CDR1 sequence selected from;

light chain CDR1 sequences differing by no more than three amino acid additions, substitutions and/or deletions from one of L1-L13 light chain CDR1 sequences;

SEQ ID NO;

28 to SEQ ID NO;

37;

(b) a light chain CDR2 sequence selected from;

light chain CDR2 sequences differing by no more than two amino acid additions, substitutions and/or deletions from one of L1-L13 light chain CDR2 sequences;

SEQ ID NO;

38 to SEQ ID NO;

45;

(c) a heavy chain CDR1 sequence selected from;

heavy chain CDR1 sequences differing by no more than two amino acid additions, substitutions and/or deletions from one of H1-H13 heavy chain CDR1 sequences;

SEQ ID NO;

6 to SEQ ID NO;

12; and

(d) a heavy chain CDR2 sequence selected from;

heavy chain CDR2 sequences differing by no more than three amino acid additions, substitutions and/or deletions from one of H1-H13 heavy chain sequences;

SEQ ID NO;

13 to SEQ ID NO;

19.

8. The antibody of claim 1, wherein the antibody is selected from murine antibodies, human antibodies, humanized antibodies, chimeric antibodies, monoclonal antibodies, polyclonal antibodies, recombinant antibodies, antigen-binding antibody fragments, single-chain antibodies, double-chain antibodies, triple-chain antibodies, tetra-chain antibodies, Fab fragments, F(fa′

)x fragments, domain antibodies, IgD antibodies, IgE antibodies, IgM antibodies, IgG1 antibodies, IgG2 antibodies, IgG3 antibodies, and IgG4 antibodies.

9. 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.

10. The GLP-1 fusion protein of claim 9, 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 hydroxyl residual terminal of the GLP-1 fusion protein polypeptide; and

N′

represents an amino residual terminal of the GLP-1 fusion protein polypeptide.

11. A polynucleotide encoding the GLP-1 fusion protein of claim 9.

12. A vector comprising the polynucleotide of claim 11.

13. A host cell comprising the vector of claim 12.

14. A pharmaceutical composition comprising the GLP-1 fusion protein of claim 9 and a pharmaceutically acceptable carrier.

15. A method of preventing or treating non-insulin-dependent diabetes in a subject, comprising administering to the subject the pharmaceutical composition of claim 14.

16. A method of preventing or treating obesity in a subject, comprising administering to the subject the pharmaceutical composition of claim 14.

17. A method of preventing or treating overweight in a subject, comprising administering to the subject the pharmaceutical composition of claim 14.

18. A method of preventing or treating overweight in a subject, comprising administering to the subject the pharmaceutical composition of claim 14.

19. A method of preventing or treating a disease responsive to GLP-1R stimulation in a subject, comprising administering to the subject the pharmaceutical composition of claim 14.

3. An antibody specifically binding to GLP-1R, comprising an amino acid sequence selected from:

(a) one or more light chain variable regions selected from;

i. light chain CDR1 sequences;

SEQ ID NO;

28 to SEQ ID NO;

37;

ii. light chain CDR2 sequences;

SEQ ID NO;

38 to SEQ ID NO;

45; and

iii. light chain CDR3 sequences;

SEQ ID NO;

46 to SEQ ID NO;

53;

(b) one or more heavy chain variable regions selected from;

i. heavy chain CDR1 sequences;

SEQ ID NO;

6 to SEQ ID NO;

12;

ii. heavy chain CDR2 sequences;

SEQ ID NO;

13 to SEQ ID NO;

19; and

iii. heavy chain CDR3 sequences;

SEQ ID NO;

20 to SEQ ID NO;

27; and

(c) a light chain variable region sequence from (a) and a heavy chain variable region sequence from (b).

4. An antibody specifically binding to GLP-1R, comprising an amino acid sequence selected from:

(a) a light chain variable domain sequence selected from;

i. an amino acid having a sequence at least 80% identical to one of L1-L13 light chain variable region sequences;

SEQ ID NO;

81, SEQ ID NO;

83, SEQ ID NO;

85, SEQ ID NO;

87, SEQ ID NO;

89, SEQ ID NO;

91, SEQ ID NO;

93, SEQ ID NO;

95, SEQ ID NO;

97, SEQ ID NO;

99, SEQ ID NO;

101, SEQ ID NO;

103, and SEQ ID NO;

105; and

ii. an amino acid sequence encoded by a polynucleotide sequence that is at least 80% identical to one of the polynucleotide sequences encoding L1-L13 light chain variable regions;

SEQ ID NO;

80, SEQ ID NO;

82, SEQ ID NO;

84, SEQ ID NO;

86, SEQ ID NO;

88, SEQ ID NO;

90, SEQ ID NO;

92, SEQ ID NO;

94, SEQ ID NO;

96, SEQ ID NO;

98, SEQ ID NO;

100, SEQ ID NO;

102, and SEQ ID NO;

104;

(b) a heavy chain variable domain sequence selected from;

i. an amino acid sequence at least 80% identical to one of H1-H13 heavy chain variable region sequences;

SEQ ID NO;

55, SEQ ID NO;

57, SEQ ID NO;

59, SEQ ID NO;

61, SEQ ID NO;

63, SEQ ID NO;

65, SEQ ID NO;

67, SEQ ID NO;

69, SEQ ID NO;

71, SEQ ID NO;

73, SEQ ID NO;

75, SEQ ID NO;

77, and SEQ ID NO;

79; and

ii. an amino acid sequence encoded by a polynucleotide sequence that is at least 80% identical to one of the polynucleotide sequences encoding H1-H13 heavy chain variable regions;

SEQ ID NO;

54, SEQ ID NO;

56, SEQ ID NO;

58, SEQ ID NO;

60, SEQ ID NO;

62, SEQ ID NO;

64, SEQ ID NO;

66, SEQ ID NO;

68, SEQ ID NO;

70, SEQ ID NO;

72, SEQ ID NO;

74, SEQ ID NO;

76, and SEQ ID NO;

78; and

(c) a light chain variable region sequence from (a) and a heavy chain variable region sequence from (b).

5. The antibody of claim 4, comprising an amino acid sequence selected from:

(a) a light chain variable region sequence selected from L1-L13 light chain variable region sequences;

SEQ ID NO;

81, SEQ ID NO;

83, SEQ ID NO;

85, SEQ ID NO;

87, SEQ ID NO;

89, SEQ ID NO;

91, SEQ ID NO;

93, SEQ ID NO;

95, SEQ ID NO;

97, SEQ ID NO;

99, SEQ ID NO;

101, SEQ ID NO;

103, and SEQ ID NO;

105;

(b) a heavy chain variable domain sequence selected from H1-H13 heavy chain variable region sequences;

SEQ ID NO;

55, SEQ ID NO;

57, SEQ ID NO;

59, SEQ ID NO;

61, SEQ ID NO;

63, SEQ ID NO;

65, SEQ ID NO;

67, SEQ ID NO;

69, SEQ ID NO;

71, SEQ ID NO;

73, SEQ ID NO;

75, SEQ ID NO;

77, and SEQ ID NO;

79; and

(c) a light chain variable region sequence from (a) and a heavy chain variable region sequence from (b).

6. The antibody of claim 5, wherein the antibody is an antibody from (c), a combination of a light chain variable region sequence from (a) and a heavy chain variable region sequence from (b), selecting from L1H1, L2H2, L3H3, L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L10H10, L11H11, L12H12, and L13H13.

7. The antibody of claim 6, 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.