Compositions and methods relating to glucagon receptor antibodies

• US 20090041784A1
• Filed: 09/20/2007
• Published: 02/12/2009
• Est. Priority Date: 09/20/2006
• Status: Active Grant

• Current Assignee: Amgen Inc.
• Original Assignee: Amgen Inc.
• Inventors: Hu, Shaw-Fen Sylvia, Boone, Thomas C., Yan, Hai, Lindberg, Richard A.
• Granted Patent: US 7,947,809 B2
• Time in Patent Office: Days
• US Class Current: 424/172.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07K 14/723 G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor
• CPC Class Codes: C07K 16/2869 against hormone receptors for antibodies against neuromediator receptors C07K16/286
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• 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 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

The present disclosure provides compositions and methods relating to antigen binding proteins, in particular, antibodies which specifically bind to the human glucagon receptor. The disclosure provides nucleic acids encoding such antigen binding proteins and antibodies and methods of making and using such antibodies including methods of treating and preventing type 2 diabetes and related disorders by administering such antibodies to a subject in need of such treatment.

Claims

1. An isolated antigen binding protein comprising an amino acid sequence selected from the group consisting of:

a. a light chain CDR3 comprising a sequence selected from the group consisting of;

i. a light chain CDR3 sequence that differs by no more than a total of three amino acid additions, substitutions, and/or deletions from a CDR3 sequence selected from the group consisting of the light chain CDR3 sequences of L1-L23, SEQ ID NOs;

72;

74, 76, 78, 80, 83, 85, 87, 89, 91, 93, 95, 97, 100;

ii. L Q X₂₁ N S X₂₂ P L T (SEQ ID NO;

     208); and

iii. Q A W D S X₂₃ T V X₂₄ (SEQ ID NO;

209);

b. a heavy chain CDR3 sequence comprising a sequence selected from the group consisting of;

i. a heavy chain CDR3 sequence that differs by no more than a total of four amino acid additions, substitutions, and/or deletions from a CDR3 sequence selected from the group consisting of the heavy chain CDR3 sequences of H1-H23, SEQ ID NOs;

165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199;

ii. E X₂₅ X₂₆ X₂₇ Y D I L T G Y X₂₈ X₂₉ Y Y G X₃₀ D V (SEQ ID NO;

     210); and

iii. X₃₁ G G G F D Y (SEQ ID NO;

211);

or

c. the light chain CDR3 sequence of (a) and the heavy chain CDR3 sequence of (b);

wherein,

X₂₁ is a histidine residue, or a glutamine residue,

X₂₂ is an asparagine residue, an aspartate residue, or a tyrosine residue,

X₂₃ is an asparagine residue or a serine residue,

X₂₄ is an isoleucine residue or a valine residue,

X₂₅ is a lysine residue, a glutamate residue, or a proline residue,

X₂₆ is an aspartate residue, a threonine residue, a glutamine residue, or a proline residue,

X₂₇ is a histidine residue or a tyrosine residue,

X₂₈ is an asparagine residue, a histidine residue, an aspartate residue, or a phenylalanine residue,

X₂₉ is a tyrosine residue, a histidine residue, or an asparagine residue,

X₃₀ is a leucine residue or a methionine residue,

X₃₁ is a leucine residue or a methionine residue,

and wherein said antigen binding protein specifically binds to the human glucagon receptor.

2. The isolated antigen binding protein of claim 1, comprising either:

a. a light chain CDR1 sequence selected from the group consisting of;

i. a light chain CDR1 that differs by no more than three amino acids additions, substitutions, and/or deletions from a CDR1 sequence of L1-L23, SEQ ID NOs;

10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38 and 41;

ii. R S X₁ Q S L L D X₂ X₃ D G T Y T L D (SEQ ID NO;

200);

iii. R A S Q X₄ I R N D X₅ G (SEQ ID NO;

201); and

iv. S G D K L G D K Y X₆ C (SEQ ID NO;

     202);

wherein

X₁ is a serine residue or a threonine residue,

X₂ is an arginine residue or a serine residue,

X₃ is an aspartate residue or an alanine residue,

X₄ is a glycine residue or an aspartate residue,

X₅ is a leucine residue or a phenylalanine residue,

X₆ is a valine residue or an alanine residue,

b. a light chain CDR2 sequence selected from the group consisting of;

i. a light chain CDR2 that differs by no more than two amino acid additions, substitutions, and/or deletions from a CDR2 sequence of L1-L23, SEQ ID NOs;

43, 45, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, and 70;

ii. A A S S L X₉ S (SEQ ID NO;

     204); and

iii. Q X₁₀ X₁₁ K R P S (SEQ ID NO;

     205);

wherein

X₉ is a glutamine residue or a glutamate residue,

X₁₀ is a serine residue or a threonine residue,

X₁₁ is a threonine residue, or a serine residue,

c. a heavy chain CDR1 sequence selected from the group consisting of;

i. a heavy chain CDR1 that differs by no more than two amino acid additions, substitutions, and/or deletions from a CDR1 sequence of H1-H23, SEQ ID NOs;

102, 104, 106, 108, 111, 113, 115, 117, 118, 120 and 122; and

ii. X₇ Y X₈ M H (SEQ ID NO;

     203);

wherein

X₇ is a serine residue or a threonine residue,

X₈ is a glycine residue or an aspartate residue; and

d. a heavy chain CDR2 selected from the group consisting of;

i. a heavy sequence that differs by no more than three amino acid additions, substitutions, and/or deletions from a CDR2 sequence of H1-H23, SEQ ID NOs;

124, 126, 128, 130, 132, 134, 136, 138, 140, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, and 163;

ii. X₁₂ I W X₁₃ D G S X₁₄ K Y Y X₁₅ D S V K G (SEQ ID NO;

     206);

iii. X₁₆ I S X₁₇ D G S X₁₈ K Y X₁₉ X₂₀ D S V K G (SEQ ID NO;

     207);

wherein

X₁₂ is a serine residue, a phenylalanine residue, a valine residue, or a glutamate residue,

X₁₃ is a tyrosine residue or an asparagine residue,

X₁₄ is an asparagine residue or a glutamate residue,

X₁₅ is a valine residue or an alanine residue,

X₁₆ is a valine residue or a phenylalanine residue,

X₁₇ is a histidine residue, an aspartate residue, or a tyrosine residue

X₁₈ is an aspartate residue, an asparagine residue, or a histidine residue,

X₁₉ is a tyrosine residue, or a serine residue,

X₂₀ is an alanine residue or a glycine residue,

e. the light chain CDR1 of (a) and the light chain CDR2 of (b);

f. the light chain CDR1 of (a) and the heavy chain CDR1 of (c);

g. the light chain CDR1 of (a) and the heavy chain CDR2 of (d);

h. the light chain CDR1 (b) and the heavy chain CDR1 of (c);

i. the heavy chain CDR1 of (c) and the heavy chain CDR2 of (d);

j. the light chain CDR2 of (b) and the heavy chain CDR2 of (d);

k. the light chain CDR1 of (a), the light chain CDR2 of (b), and the heavy chain CDR1 of (c);

l. the light chain CDR2 of (b), the heavy CDR1 of (c), and the heavy chain CDR2 of (d);

m. the light chain CDR1 of (a), the heavy chain CDR1 of (c), and the heavy chain CDR2 of (d);

or

n. the light chain CDR1 of (a), the light chain CDR2 of (b), the heavy chain CDR2 of (c), and the heavy chain CDR2 of (d),

wherein said antigen binding protein specifically binds to the human glucagon receptor.

3. The isolated antigen binding protein of claim 1, comprising either:

a. a light chain variable domain comprising;

i. a light chain CDR1 sequence selected from SEQ ID NOs;

10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, and 41;

ii. a light chain CDR2 sequence selected from SEQ ID NOs;

43, 45, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, and 70;

iii. a light chain CDR3 sequence selected from SEQ ID NOs;

72, 74, 76, 78, 80, 83, 85, 87, 89, 91, 93, 95, 97, and 100; and

b. a heavy chain variable domain comprising;

i. a heavy chain CDR1 sequence selected from SEQ ID NOs;

102, 104, 106, 108, 111, 113, 115, 117, 118, 120, and 122;

ii. a heavy chain CDR2 sequence selected from SEQ ID NOs;

124, 126, 128, 130, 132, 134, 136, 138, 140, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, and 163; and

iii. a heavy chain CDR3 sequence selected from SEQ ID NOs;

165, 167, 169, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, and 199;

or

c. the light chain variable domain of (a) and the heavy chain variable domain of (b), wherein the antigen binding protein specifically binds to human glucagon receptor.

4. The isolated antigen binding protein of claim 1 comprising either:

a. a light chain variable domain sequence selected from the group consisting of;

i. amino acids having a sequence at least 80% identical to a light chain variable domain sequence selected from L1-L23, SEQ ID NOs;

213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, and 257;

ii. a sequence of amino acids encoded by a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence encoding the light chain variable domain sequence of L1-L23, SEQ ID NOs;

212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, and 256;

iii. a sequence of amino acids encoded by a polynucleotide sequence that hybridizes under moderately stringent conditions to the complement of a polynucleotide consisting of a light chain variable domain sequence of L1-L23 of SEQ ID NOs;

212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, and 256;

b. a heavy chain variable domain sequence selected from the group consisting of;

i. a sequence of amino acids that is at least 80% identical to a heavy chain variable domain sequence of H1-H23 of SEQ ID NOs;

259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, and 303;

ii. a sequence of amino acids encoded by a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence encoding the heavy chain variable domain sequence of H1-H23, SEQ ID NOs;

258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 292, 294, 296, 298, 300, and 302;

iii. a sequence of amino acids encoded by a polynucleotide sequence that hybridizes under moderately stringent conditions to the complement of a polynucleotide consisting of a heavy chain variable domain sequence of H1-H23, SEQ ID NOs;

258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 292, 294, 296, 298, 300, and 302;

or

c. the light chain variable domain of (a) and the heavy chain variable domain of (b), wherein the antigen binding protein binds to the human glucagon receptor.

5. The isolated antigen binding protein of claim 4, comprising either:

a. a light chain variable domain sequence selected from the group consisting of;

L1-L23 of SEQ ID NOs;

213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241. 243, 245, 247, 249, 251, 253, 255, and 257;

b. a heavy chain variable domain sequence selected from the group consisting of;

H1-H23 of SEQ ID NOs;

259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, and 303;

or

c. the light chain variable domain of (a) and the heavy chain variable domain of (b), wherein the antigen binding protein specifically binds to the human glucagon receptor.

6. The antigen binding protein of claim 5, wherein the light chain variable domain and a heavy chain variable domain are selected from the group of combinations consisting of:

L1H1, L1H1, L2H2, L3H3, L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L10H10, L11H11, L12H12, L13H13, L14H14, L15H15, L16H16, L17H17, L18H18, L19H19, L20H20, L21H21, L22H22, and L23H23,

wherein the antigen binding protein specifically binds to the human glucagon receptor.

7. The antigen binding protein of claim 6, further comprising:

a. the light chain constant sequence of SEQ ID NO;

305;

b. the light chain constant sequence of SEQ ID NO;

307;

c. the heavy chain constant sequence of SEQ ID NO;

309;

d. the light chain constant sequence of SEQ ID NO;

305 and the heavy chain constant sequence of SEQ ID NO;

309, or

e. the light chain constant sequence of SEQ ID NO;

307 and the heavy chain constant sequence of SEQ ID NO;

309.

8. The isolated binding protein of claim 1, wherein the antigen binding protein is selected from the group consisting of a human antibody, a humanized antibody, chimeric antibody, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, an antigen-binding antibody fragment, a single chain antibody, a diabody, a triabody, a tetrabody, a Fab fragment, an F(fa′

)x fragment, a domain antibody, an IgD antibody, an IgE antibody, and IgM antibody, and IgG1 antibody, and IgG2 antibody, and IgG3 antibody, and IgG4 antibody, and IgG4 antibody having at least one mutation in the hinge region.

9. The antigen binding protein of claim 1, that, when bound to the human glucagon receptor:

a. binds to the human glucagon receptor with substantially the same Kd as a reference antibody;

b. inhibits glucagon stimulation of the human glucagon receptor with substantially the same IC₅₀ as said reference antibody;

or

c. cross-competes for binding with said reference antibody on the human glucagon receptor,

wherein said reference antibody comprises a combination of light chain and heavy chain variable domain sequences selected from the group consisting of L1H1, L2H2, L3H3, L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L11H11, L12H12, L13H13, L15H15, L21H21, and L22H22.

12. A pharmaceutical composition comprising the antigen binding protein of claim 1 in admixture with a pharmaceutically acceptable carrier thereof.

14. An isolated nucleic acid comprising a polynucleotide sequence encoding the light chain variable domain, the heavy chain variable domain, or both, of the antigen binding protein of claim 4.

15. The isolated nucleic acid of claim 14, wherein the sequence is selected from L1-L23, SEQ ID NOs:

212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, and 256;

H1-H23, SEQ ID NOs;

258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, and 302, or both.

16. An expression vector comprising the nucleic acid of claim 14.

17. An isolated cell comprising the nucleic acid of claim 14.

18. The isolated cell of claim 17, wherein the chromosome of the cell comprises the nucleic acid.

19. The isolated cell of claim 18, wherein the cell is a hybridoma.

20. The isolated cell of claim 17, wherein an expression vector comprises said nucleic acid.

21. A method of producing an antigen binding protein that specifically binds to the human glucagon receptor comprising incubating the host cell of claim 17 under conditions that allow it to express the antigen binding protein.

22. A method of preventing or treating a condition in a subject in need of such treatment comprising administering a therapeutically effective amount of the composition of claim 12 to the subject, wherein the condition is treatable by lowering blood glucose.

23. The method of claim 22, wherein the condition is selected from hyperglycemia, hyperinsulinemia, impaired fasting glucose, impaired glucose tolerance, dyslipodemia, and type 2 diabetes.

10. An isolated human antibody that, when bound to the human glucagon receptor:

a. binds to the human glucagon receptor with substantially the same Kd as a reference antibody;

b. inhibits glucagon stimulation of the human glucagon receptor with substantially the same IC₅₀ as said reference antibody;

or

c. cross-competes for binding with said reference antibody on human glucagon receptor,

wherein said reference antibody is selected from the group consisting of A-1, A-2, A-3, A-4, A-5, A-6, A-7, A-8, A-9, A-11, A-12, A-13, A-15, A-21, and A-22.

11. The isolated human antibody of claim 10, that, when bound to the human glucagon receptor:

a. specifically binds to amino acids Ser80 to Ser119 of the human glucagon receptor;

b. reduces glucagon signalling with an IC50 value of 90 nM or less;

c. lowers blood glucose in an animal model;

or

d. (a) and (b), or

e. (a), (b) and (c).

13. A pharmaceutical composition comprising the human antibody of claim 10 in admixture with a pharmaceutically acceptable carrier thereof.

24. A method of preventing or treating a condition in a subject in need of such treatment comprising administering a therapeutically effective amount of the composition of claim 13 to the subject, wherein the condition is treatable by lowering blood glucose.

25. The method of claim 24, wherein the condition is selected from hyperglycemia, hyperinsulinemia, impaired fasting glucose, impaired glucose tolerance, dyslipodemia, and type 2 diabetes.