Humanized antibody molecules specific for IL-31

• US 8,470,979 B2
• Filed: 12/08/2008
• Issued: 06/25/2013
• Est. Priority Date: 12/07/2007
• Status: Active Grant

• Current Assignee: Zymogenetics Incorporated (Bristol-Myers Squibb Company)
• Original Assignee: Zymogenetics Incorporated (Bristol-Myers Squibb Company), Merck Serono SA (Merck & Co., Inc.)
• Inventors: Beckmann, Roland, Bondensgaard, Kent
• Primary Examiner(s): KEMMERER, ELIZABETH
• Application Number:
  US12/329,820
• Publication Number:
  US 20090208494A1
• Time in Patent Office: 1,660 Days
• Field of Search: None
• US Class Current: 530/388.23
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/04 Antipruritics
• CPC Class Codes: A61P 17/14 for baldness or alopecia
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/244 Interleukins [IL]
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/51 Complete heavy chain or Fd fragment, i.e. VH + CH1
• CPC Class Codes: C07K 2317/515 Complete light chain, i.e. VL + CL
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• 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
• CPC Class Codes: C12N 15/63 Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
• CPC Class Codes: C12N 5/10 Cells modified by introduction of foreign genetic material

Abstract

The invention provides humanized mouse anti-human IL-31 antibodies and antibody fragments that are capable of binding IL-31 and thereby neutralizing, inhibiting, limiting, or reducing the proinflammatory or pro-pruritic effects of IL-31.

Claims

1. An isolated antibody or antibody fragment that binds to human IL-31, comprising

a) a humanized heavy chain variable domain comprising CDR1, CDR2, and CDR3 consisting of amino acid sequences SEQ ID NO:

1, 2 and 3, respectively or consisting of amino acid sequences SEQ ID NO;

1, 4 and 3, respectively; and

b) a humanized light chain variable domain comprising CDR1, CDR2, and CDR3 consisting of amino acid sequences of SEQ ID NO;

5, 6 and 7, respectively.

2. The antibody or antibody fragment according to claim 1, wherein

a) said humanized heavy chain variable domain comprises framework regions FR1, FR2, FR3 and FR4 and wherein said humanized heavy chain variable domain is selected from the group consisting of:

1) a heavy chain variable domain wherein the FR1 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

8, the FR2 sequence has at least 90% identity to SEQ ID NO;

9, the FR3 sequence has at least 90% identity to SEQ ID NO;

10 and the FR4 sequence has at least 90% identity to SEQ ID NO;

11 ;

2) a heavy chain variable domain wherein the FR1 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

12, the FR2 sequence has at least 90% identity to SEQ ID NO;

13, the FR3 sequence has at least 90% identity to SEQ ID NO;

14 and the FR4 sequence has at least 90% identity to SEQ ID NO;

15 ;

3) a heavy chain variable domain wherein the FR1 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

12, the FR2 sequence has at least 90% identity to SEQ ID NO;

13, the FR3 sequence has at least 90% identity to SEQ ID NO;

16 and the FR4 sequence has at least 90% identity to SEQ ID NO;

15 ;

wherein the amino acid at position 29 in SEQ ID NO;

8 or SEQ ID NO;

12 is leucine and the amino acid at position 32 SEQ ID NO;

14 or SEQ ID NO;

16 is phenylalanine; and

b) said humanized light chain variable domain comprises framework regions FR5, FR6, FR7, and FR8, wherein said humanized heavy chain variable region is selected from the group consisting of;

1) a light chain variable domain wherein the FR5 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

17, the FR6 sequence has at least 90% identity to SEQ ID NO;

18, the FR7 sequence has at least 90% identity to SEQ ID NO;

19 and the FR8 sequence has at least 90% identity to SEQ ID NO;

20; and

2) a light chain variable domain wherein the FR5 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

17, the FR6 sequence has at least 90% identity to SEQ ID NO;

18, the FR7 sequence has at least 90% identity to SEQ ID NO;

21 and the FR8 sequence has at least 90% identity to SEQ ID NO;

20, and

wherein the amino acid at position 15 in SEQ ID NO;

19 or SEQ ID NO;

21 is tyrosine.

3. The antibody or antibody fragment according to claim 2, wherein the FR1 sequence consists of the amino acid sequence of SEQ ID NO:

8, the FR2 sequence consists of the amino acid sequence of SEQ ID NO;

9, the FR3 sequence consists of the amino acid sequence of SEQ ID NO;

10 and the FR4 sequence consists of the amino acid sequence of SEQ ID NO;

11 and wherein the FR5 sequence consists of the amino acid sequence of SEQ ID NO;

17, the FR6 sequence consists of the amino acid sequence of SEQ ID NO;

18, the FR7 sequence consists of the amino acid sequence of SEQ ID NO;

19 and the FR8 sequence consists of the amino acid sequence of SEQ ID NO;

20.

4. The antibody or antibody fragment according to claim 2, wherein the FR1 sequence consists of the amino acid sequence of SEQ ID NO:

12, the FR2 sequence consists of the amino acid sequence of SEQ ID NO;

13, the FR3 sequence consists of the amino acid sequence of SEQ ID NO;

14 and the FR4 sequence consists of the amino acid sequence of SEQ ID NO;

15 and wherein the FR5 sequence consists of the amino acid sequence of SEQ ID NO;

17, the FR6 sequence consists of the amino acid sequence of SEQ ID NO;

18, the FR7 sequence consists of the amino acid sequence of SEQ ID NO;

19 and the FR8 sequence consists of the amino acid sequence of SEQ ID NO;

20.

5. The antibody or antibody fragment according to claim 2, wherein the FR1 sequence consists of the amino acid sequence of SEQ ID NO:

12, the FR2 sequence consists of the amino acid sequence of SEQ ID NO;

13, the FR3 sequence consists of the amino acid sequence of SEQ ID NO;

14 and the FR4 sequence consists of the amino acid sequence of SEQ ID NO;

15 and wherein the FR5 sequence consists of the amino acid sequence of SEQ ID NO;

17, the FR6 sequence consists of the amino acid sequence of SEQ ID NO;

18, the FR7 sequence consists of the amino acid sequence of SEQ ID NO;

21 and the FR8 sequence consists of the amino acid sequence of SEQ ID NO;

20.

6. The antibody or antibody fragment according to claim 2, wherein the amino acid at position 8 in SEQ ID NO:

14 or SEQ ID NO;

16 is lysine.

7. The antibody or antibody fragment according to claim 1, wherein the antibody comprises a heavy chain immunoglobulin constant domain selected from the group consisting of:

a) a human IgG1 constant domain;

b) a human IgG2 constant domain;

c) a human IgG3 constant domain;

d) a human IgG4 constant domain;

e) a human IgM constant domain;

f) a human IgE constant domain; and

g) a human IgA constant domain.

8. The antibody fragment according to claim 1, wherein said fragment is selected from the group consisting of:

Fab, Fab′

, F(ab′

)2, Fv fragments, scFv;

diabodies;

linear antibodies;

single-chain antibody molecules;

monobodies; and

multispecific antibodies formed from antibody fragments.

9. The antibody or antibody fragment according to claim 1, wherein said antibody or antibody fragment comprises PEG.

10. A pharmaceutical composition comprising the antibody or antibody fragment of claim 1.

23. The antibody or antibody fragment according to claim 2, wherein the FR1 sequence consists of the amino acid sequence of SEQ ID NO:

12, the FR2 sequence consists of the amino acid sequence of SEQ ID NO;

13, the FR3 sequence consists of the amino acid sequence of SEQ ID NO;

16 and the FR4 sequence consists of the amino acid sequence of SEQ ID NO;

15, and wherein the FR5 sequence consists of the amino acid sequence of SEQ ID NO;

17, the FR6 sequence consists of the amino acid sequence of SEQ ID NO;

18, the FR7 sequence consists of the amino acid sequence of SEQ ID NO;

21 and the FR8 sequence consists of the amino acid sequence of SEQ ID NO;

20.

24. The antibody or antibody fragment according to claim 1, wherein the humanized heavy chain variable domain has at least 90% sequence identity to the amino acid sequence of SEQ ID NO:

47 and the humanized light chain variable domain has at least 90% sequence identity to the amino acid sequence of SEQ ID NO;

46.

25. The antibody or antibody fragment according to claim 24, wherein the antibody comprises a heavy chain immunoglobulin constant domain selected from the group consisting of:

a) a human IgG1 constant domain;

b) a human IgG2 constant domain;

c) a human IgG3 constant domain;

d) a human IgG4 constant domain;

e) a human IgM constant domain;

f) a human IgE constant domain; and

g) a human IgA constant domain.

26. The antibody fragment according to claim 24, wherein said fragment is selected from the group consisting of:

Fab, Fab′

, F(ab′

)2, Fv fragments, scFv;

diabodies;

linear antibodies;

single-chain antibody molecules;

monobodies; and

multispecific antibodies formed from antibody fragments.

27. The antibody or antibody fragment according to claim 24, wherein said antibody or antibody fragment comprises PEG.

28. A pharmaceutical composition comprising the antibody or antibody fragment of claim 24.

29. The antibody or antibody fragment according to claim 1, wherein the humanized heavy chain variable domain has at least 90% sequence identity to the amino acid sequence of SEQ ID NO:

44 and the humanized light chain variable domain has at least 90% sequence identity to the amino acid sequence of SEQ ID NO;

26.

30. The antibody or antibody fragment according to claim 29, wherein the antibody comprises a heavy chain immunoglobulin constant domain selected from the group consisting of:

a) a human IgG1 constant domain;

b) a human IgG2 constant domain;

c) a human IgG3 constant domain;

d) a human IgG4 constant domain;

e) a human IgM constant domain;

f) a human IgE constant domain; and

g) a human IgA constant domain.

31. The antibody fragment according to claim 29, wherein said fragment is selected from the group consisting of:

Fab, Fab′

, F(ab′

)2, Fv fragments, scFv;

diabodies;

linear antibodies;

single-chain antibody molecules;

monobodies; and

multispecific antibodies formed from antibody fragments.

32. The antibody or antibody fragment according to claim 29, wherein said antibody or antibody fragment comprises PEG.

33. A pharmaceutical composition comprising the antibody or antibody fragment of claim 29.

39. The antibody or antibody fragment according to claim 2, wherein the FR1 sequence consists of the amino acid sequence of SEQ ID NO:

8, the FR2 sequence consists of the amino acid sequence of SEQ ID NO;

9, the FR3 sequence consists of the amino acid sequence of SEQ ID NO;

10, and the FR4 sequence consists of the amino acid sequence of SEQ ID NO;

11, and wherein the FR5 sequence consists of the amino acid sequence of SEQ ID NO;

17, the FR6 sequence consists of the amino acid sequence of SEQ ID NO;

18, the FR7 sequence consists of the amino acid sequence of SEQ ID NO;

21, and the FR8 sequence consists of the amino acid sequence of SEQ ID NO;

20.

40. The antibody or antibody fragment according to claim 2, wherein the FR1 sequence consists of the amino acid sequence of SEQ ID NO:

12, the FR2 sequence consists of the amino acid sequence of SEQ ID NO;

13, the FR3 sequence consists of the amino acid sequence of SEQ ID NO;

16, and the FR4 sequence consists of the amino acid sequence of SEQ ID NO;

15, and wherein the FR5 sequence consists of the amino acid sequence of SEQ ID NO;

17, the FR6 sequence consists of the amino acid sequence of SEQ ID NO;

18, the FR7 sequence consists of the amino acid sequence of SEQ ID NO;

19, and the FR8 sequence consists of the amino acid sequence of SEQ ID NO;

20.

11. An isolated antibody comprising a light chain comprising the amino acid sequence of SEQ ID NO:

46 and a heavy chain comprising the amino acid sequence of SEQ ID NO;

47.

12. An isolated antibody or antibody fragment that binds to human IL-31, comprising:

a) a humanized heavy chain variable domain comprising a CDR1 consisting of the amino acid sequence of SEQ ID NO;

1, a CDR3 consisting of the amino acid sequence of SEQ ID NO;

3, and a CDR2 consisting of the amino acid sequence of AIYPGDGDTRYSXaa1Xaa2FXaa3G (SEQ ID NO;

     22) wherein Xaa1 is glutamine or proline, Xaa2 is serine or lysine and Xaa3 is glutamine or lysine and wherein said humanized heavy chain variable domain is selected from the group consisting of;

1) a heavy chain variable domain wherein the FR1 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

8, the FR2 sequence has at least 90% identity to SEQ ID NO;

9, the FR3 sequence has at least 90% identity to SEQ ID NO;

10 and the FR4 sequence has at least 90% identity to SEQ ID NO;

11 ;

2) a heavy chain variable domain wherein the FR1 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

12, the FR2 sequence has at least 90% identity to SEQ ID NO;

13, the FR3 sequence has at least 90% identity to SEQ ID NO;

14 and the FR4 sequence has at least 90% identity to SEQ ID NO;

15; and

3) a heavy chain variable domain wherein the FR1 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

12, the FR2 sequence has at least 90% identity to SEQ ID NO;

13, the FR3 sequence has at least 90% identity to SEQ ID NO;

16 and the FR4 sequence has at least 90% identity to SEQ ID NO;

15 ;

and wherein the amino acid at position 29 in SEQ ID NO;

8 or SEQ ID NO;

12 is leucine and amino acid at position 32 in SEQ ID NO;

14 or SEQ ID NO;

16 is phenylalanine; and

b) a humanized light chain variable domain comprising a CDR1 consisting of the amino acid sequence of SEQ ID NO;

5, a CDR2 consisting of the amino acid sequence of SEQ ID NO;

6, and a CDR3 consisting of the amino acid sequence of SEQ ID NO;

7, and wherein said humanized heavy chain variable domain is selected from the group consisting of;

1) a light chain variable domain wherein the FR5 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

17, the FR6 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

18, the FR7 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

19, and the FR8 sequence has at least 90% sequence identity to the amino acid sequence of SEQ ID NO;

20; and

2) a light chain variable domain wherein the FR5 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

17, the FR6 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

18, the FR7 sequence has at least 90% identity to the amino acid sequence of SEQ ID NO;

21, and the FR8 sequence has at least 90% sequence identity to the amino acid sequence of SEQ ID NO;

20;

wherein the amino acid at position 15 in SEQ ID NO;

19 or SEQ ID NO;

21 is tyrosine.

13. The antibody or antibody fragment according to claim 12, wherein Xaa1, Xaa2, and Xaa3 of SEQ ID NO:

22 are selected from the group consisting of;

a) Xaa1 glutamine, Xaa2 is lysine, and Xaa3 is lysine;

b) Xaa1 is proline, Xaa2 is serine, and Xaa3 is glutamine; and

c) Xaa1 is glutamine, Xaa2 is lysine, and Xaa3 is glutamine.

14. The antibody or antibody fragment according to claim 12, wherein the amino acid at position 8 in SEQ ID NO:

14 or SEQ ID NO;

16 is leucine.

15. The antibody or antibody fragment according to claim 12, wherein the antibody comprises a heavy chain immunoglobulin constant domain selected from the group consisting of:

a) a human IgG1 constant domain;

b) a human IgG2 constant domain;

c) a human IgG3 constant domain;

d) a human IgG4 constant domain;

e) a human IgM constant domain;

f) a human IgE constant domain; and

g) a human IgA constant domain.

16. The antibody fragment according to claim 12, wherein said fragment is selected from the group consisting of:

Fab, Fab′

, F(ab′

)2, Fv fragments, scFv;

diabodies;

linear antibodies;

single-chain antibody molecules;

monobodies; and

multispecific antibodies formed from antibody fragments.

17. The antibody or antibody fragment according to claim 12, wherein said antibody or antibody fragment comprises PEG.

18. A pharmaceutical composition comprising the antibody or antibody fragment of claim 12.

19. An isolated antibody or antibody fragment that binds to human IL-31, comprising a humanized heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:

44 and a humanized light chain variable domain comprising the amino acid sequence of SEQ ID NO;

26.

20. The antibody or antibody fragment according to claim 19, wherein the antibody comprises a heavy chain immunoglobulin constant domain selected from the group consisting of:

a) a human IgG1 constant domain;

b) a human IgG2 constant domain;

c) a human IgG3 constant domain;

d) a human IgG4 constant domain;

e) a human IgM constant domain;

f) a human IgE constant domain; and

g) a human IgA constant domain.

21. The antibody fragment according to claim 19, wherein said fragment is selected from the group consisting of:

Fab, Fab′

, F(ab′

)2, Fv fragments, scFv;

diabodies;

linear antibodies;

single-chain antibody molecules;

monobodies; and

multispecific antibodies formed from antibody fragments.

22. The antibody or antibody fragment according to claim 19, wherein said antibody or antibody fragment comprises PEG.

34. An isolated antibody or antibody fragment that binds to human IL-31, comprising

a) a humanized heavy chain variable domain comprising:

i) a FR1 sequence comprising the amino acid sequence of SEQ ID NO;

8 or of SEQ ID NO;

12;

ii) a variable heavy domain CDR1 sequence comprising the amino acid sequence of SEQ ID NO;

1;

iii) a FR2 sequence comprising the amino acid sequence of SEQ ID NO;

9 or of SEQ ID NO;

13;

iv) a variable heavy domain CDR2 sequence comprising the amino acid sequence of SEQ ID NO;

2 or of SEQ ID NO;

4;

v) a FR3 sequence comprising the amino acid sequence of SEQ ID NO;

10, of SEQ ID NO;

14, or of SEQ ID NO;

16;

vi) a variable heavy domain CDR3 sequence comprising the amino acid sequence of SEQ ID NO;

3; and

vii) a FR4 sequence comprising the amino acid sequence of SEQ ID NO;

11 or of SEQ ID NO;

15; and

b) a humanized light chain variable domain comprising;

i) a FR5 sequence comprising the amino acid sequence of SEQ ID NO;

17;

ii) a variable light domain CDR1 sequence comprising the amino acid sequence of SEQ ID NO;

5;

iii) a FR6 sequence comprising the amino acid sequence of SEQ ID NO;

18;

iv) a variable light domain CDR2 sequence comprising the amino acid sequence of SEQ ID NO;

6;

v) a FR7 sequence comprising the amino acid sequence of SEQ ID NO;

19 or of SEQ ID NO;

21;

vi) a variable light domain CDR3 sequence comprising the amino acid sequence of SEQ ID NO;

7; and

vii) a FR8 sequence comprising the amino acid sequence of SEQ ID NO;

20.

35. The antibody or antibody fragment according to claim 34, wherein the antibody comprises a heavy chain immunoglobulin constant domain selected from the group consisting of:

a) a human IgG1 constant domain;

b) a human IgG2 constant domain;

c) a human IgG3 constant domain;

d) a human IgG4 constant domain;

e) a human IgM constant domain;

f) a human IgE constant domain; and

g) a human IgA constant domain.

36. The antibody fragment according to claim 34, wherein said fragment is selected from the group consisting of:

Fab, Fab′

, F(ab′

)2, Fv fragments, scFv;

diabodies;

linear antibodies;

single-chain antibody molecules;

monobodies; and

multispecific antibodies formed from antibody fragments.

37. The antibody or antibody fragment according to claim 34, wherein said antibody or antibody fragment comprises PEG.

38. A pharmaceutical composition comprising the antibody or antibody fragment of claim 34.