COMPOSITIONS FOR INHIBITING MASP-2 DEPENDENT COMPLEMENT ACTIVATION

• US 20120282263A1
• Filed: 05/04/2012
• Published: 11/08/2012
• Est. Priority Date: 05/04/2011
• Status: Active Grant

• Current Assignee: Omeros Corporation
• Original Assignee: Omeros Corporation
• Inventors: Dudler, Thomas, Gombotz, Wayne R., Tedford, Clark E., Kavlie, Anita, Hagemann, Urs Beat, Reiersen, Herald, Parent, James Brian, Kiprijanov, Sergej
• Granted Patent: US 9,011,860 B2
• Time in Patent Office: Days
• US Class Current: 424/142.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/53 Hinge
• CPC Class Codes: C07K 2317/54 F(ab')2
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• 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 2317/622 Single chain antibody (scFv)
• 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 9/6424 Serine endopeptidases (3.4.21)
• CPC Class Codes: C12Y 304/21104 Mannan-binding lectin-associated serine protease-2 (3.4.21.104)

Abstract

The present invention relates to anti-MASP-2 inhibitory antibodies and compositions comprising such antibodies for use in inhibiting the adverse effects of MASP-2 dependent complement activation.

Claims

1. An isolated human monoclonal antibody, or antigen binding fragment thereof, that binds to human MASP-2, comprising:

(i) a heavy chain variable region comprising CDR-H1, CDR-H2 and CDR-H3 sequences; and

(ii) a light chain variable region comprising CDR-L1, CDR-L2 and CDR-L3, wherein the heavy chain variable region CDR-H3 sequence comprises an amino acid sequence set forth as SEQ ID NO;

38 or SEQ ID NO;

90, and conservative sequence modifications thereof, wherein the light chain variable region CDR-L3 sequence comprises an amino acid sequence set forth as SEQ ID NO;

51 or SEQ ID NO;

94, and conservative sequence modifications thereof, and wherein the isolated antibody inhibits MASP-2 dependent complement activation.

2. The isolated antibody or antigen binding fragment of claim 1, wherein the heavy chain variable region CDR-H2 sequence comprises an amino acid sequence set forth as SEQ ID NO:

32 or 33, and conservative sequence modifications thereof.

3. The isolated antibody or antigen binding fragment of claim 2, wherein the heavy chain variable region CDR-H1 sequence comprises an amino acid sequence set forth as SEQ ID NO:

28 or SEQ ID NO;

29, and conservative modifications thereof.

4. The isolated antibody or antigen binding fragment of claim 3, wherein the light chain variable region CDR-L2 sequence comprises an amino acid sequence set forth as SEQ ID NO:

93 and conservative modifications thereof.

5. The isolated antibody or antigen binding fragment of claim 4, wherein the light chain variable region CDR-L1 sequence comprises an amino acid sequence set forth as SEQ ID NO:

91 or SEQ ID NO;

92 an conservative modifications thereof.

6. The isolated antibody or antigen binding fragment of claim 3, wherein the CDR-H1 of the heavy chain variable region comprises SEQ ID NO:

28.

7. The isolated antibody or antigen binding fragment of claim 2, wherein the CDR-H2 of the heavy chain variable region comprises SEQ ID NO:

32.

8. The isolated antibody or antigen binding fragment of claim 1, wherein the CDR-H3 of the heavy chain variable region comprises SEQ ID NO:

90.

9. The isolated antibody of claim 8, wherein the amino acid sequence set forth in SEQ ID NO:

90 contains an R at position 8.

10. The isolated antibody of claim 5, wherein the CDR-L1 of the light chain variable region comprises SEQ ID NO:

91.

11. The isolated antibody of claim 10, wherein the amino acid sequence set forth in SEQ ID NO:

91 contains an E at position 2.

12. The isolated antibody of claim 10, wherein the amino acid sequence set forth in SEQ ID NO:

91 contains a Y at position 8.

13. The isolated antibody of claim 4, wherein CDR-L2 of the light chain variable region comprises SEQ ID NO:

93, and wherein the amino acid sequence set forth in SEQ ID NO;

93 contains a K at position 2.

14. The isolated antibody of claim 4, wherein CDR-L2 of the light chain variable region comprises SEQ ID NO:

93, and wherein the amino acid sequence set forth in SEQ ID NO;

93 contains a Q at position 3.

15. The isolated antibody of claim 1, wherein the CDR-L3 of the light chain variable region comprises SEQ ID NO:

51.

16. The isolated antibody of claim 1, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of a Fab, a Fab′

fragment, a F(ab′

)2 fragment and a whole antibody.

17. The isolated antibody of claim 1, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of a single chain antibody, an ScFv, and a univalent antibody lacking a hinge region.

18. The isolated antibody of claim 1, wherein said antibody binds human MASP-2 with a K_D of 10 nM or less.

19. The antibody of claim 1, wherein said antibody inhibits C4 activation in an in vitro assay in 1% human serum at an IC₅₀ of 10 nM or less.

20. The antibody of claim 1, wherein said antibody inhibits C4 activation in 90% human serum with an IC₅₀ of 30 nM or less.

45. A nucleic acid molecule encoding the amino acid sequence of an anti-MASP-2 antibody, or fragment thereof, as set forth claim 1, 21, 41 or 43.

46. An expression cassette comprising a nucleic acid molecule encoding an anti-MASP-2 antibody of the invention according to claim 45.

47. A cell comprising at least one of the nucleic acid molecules encoding an anti-MASP-2 antibody of the invention according to claim 45 or claim 46.

48. A method of generating an isolated anti-MASP-2 antibody comprising culturing the cell of claim 47 under conditions allowing for expression of the nucleic acid molecules encoding the anti-MASP-2 antibody and isolating said anti-MASP-2 antibody.

51. A composition comprising the fully human monoclonal anti-MASP-2 antibody, or fragment thereof, of claim 1, 21, 41, 43 or 49 and a pharmaceutically acceptable excipient.

52. The composition of claim 51, wherein the composition is formulated for systemic delivery.

53. The composition of claim 52, wherein the composition is formulated for intra-arterial, intravenous, intracranial, intramuscular, inhalational, nasal or subcutaneous administration.

54. The composition of claim 52, wherein the composition is formulated for subcutaneous administration.

55. A method of inhibiting MASP-2 dependent complement activation in a human subject comprising administering a human monoclonal antibody of claim 1, 21, 41, 43 or 49 in an amount sufficient to inhibit MASP-2 dependent complement activation in said human subject.

21. An isolated human monoclonal antibody, or antigen binding fragment thereof, that binds human MASP-2, wherein the antibody comprises:

I) a) a heavy chain variable region comprising;

i) a heavy chain CDR-H1 comprising the amino acid sequence from 31-35 of SEQ ID NO;

20; and

ii) a heavy chain CDR-H2 comprising the amino acid sequence from 50-65 of SEQ ID NO;

20; and

iii) a heavy chain CDR-H3 comprising the amino acid sequence from 95-102 of either SEQ ID NO;

18 or SEQ ID NO;

20; and

b) a light chain variable region comprising;

i) a light chain CDR-L1 comprising the amino acid sequence from 24-34 of either SEQ ID NO;

22 or SEQ ID NO;

24; and

ii) a light chain CDR-L2 comprising the amino acid sequence from 50-56 of either SEQ ID NO;

22 or SEQ ID NO;

24; and

iii) a light chain CDR-L3 comprising the amino acid sequence from 89-97 of either SEQ ID NO;

22 or SEQ ID NO;

24;

or

II) a variant thereof, that is otherwise identical to said variable domains, except for up to a combined total of 10 amino acid substitutions within said CDR regions of said heavy chain and up to a combined total of 10 amino acid substitutions within said CDR regions of said light chain variable region,

wherein the antibody or variant thereof inhibits MASP-2 dependent complement activation.

22. The antibody of claim 21, wherein said antibody binds human MASP-2 with a K_D of 10 nM or less.

23. The antibody of claim 21, wherein said antibody binds an epitope in the CCP1 domain of MASP-2.

24. The antibody of claim 21, wherein said antibody inhibits C3b deposition in an in vitro assay in 1% human serum at an IC₅₀ of 10 nM or less.

25. The antibody of claim 21, wherein said antibody inhibits C3b deposition in 90% human serum with an IC₅₀ of 30 nM or less.

26. The antibody of claim 21, wherein the antibody is an antibody fragment selected from the group consisting of Fv, Fab, Fab′

, F(ab)₂ and F(ab′

)₂.

27. The antibody of claim 21, wherein the antibody is a single chain molecule.

28. The antibody of claim 21, wherein said antibody is an IgG2 molecule.

29. The antibody of claim 21, wherein said antibody is an IgG1 molecule.

30. The antibody of claim 21, wherein said antibody is an IgG4 molecule.

31. The antibody of claim 30, wherein the IgG4 molecule comprises a S228P mutation.

32. The antibody of claim 21, wherein the antibody does not substantially inhibit the classical pathway.

33. The antibody of claim 21, wherein the heavy chain variable region comprises SEQ ID NO:

20, or a variant thereof comprising at least 80% identity to SEQ ID NO;

20.

34. The antibody of claim 21, wherein the heavy chain variable region comprises SEQ ID NO:

18, or a variant thereof comprising at least 80% identity to SEQ ID NO;

18.

35. The antibody of claim 21, wherein the light chain variable region comprises SEQ ID NO:

24, or a variant thereof comprising at least 80% identity to SEQ ID NO;

24.

36. The antibody of claim 21, wherein the light chain variable region comprises SEQ ID NO:

22, or a variant thereof comprising at least 80% identity to SEQ ID NO;

22.

37. The antibody of claim 21, wherein said variant is otherwise identical to said heavy chain variable region except for up to a combined total of 1, 2, 3, 4, 5 or 6 amino acid substitutions within said CDR regions of said heavy chain variable region.

38. The antibody of claim 21, wherein said variant comprises an amino acid substitution at one or more positions selected from the group consisting of position 31, 32, 33, 34, 35, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 95, 96, 97, 98, 99, 100 or 102 of said heavy chain variable region.

39. The antibody of claim 21, wherein said variant is otherwise identical to said light chain variable region except for up to a combined total of 1, 2, 3, 4, 5, or 6 amino acid substitutions within said CDR regions of said light chain variable region.

40. The antibody of claim 21, wherein said variant comprises an amino acid substitution at one or more positions selected from the group consisting of position 25, 26, 27, 29, 31, 32, 33, 51, 52, 89, 92, 93, 95, 96 or 97 of said light chain variable region.

41. An isolated monoclonal antibody, or antigen-binding fragment thereof, that binds to human MASP-2, comprising a heavy chain variable region comprising any one of the amino acid sequences set forth in SEQ ID NO:

18, SEQ ID NO;

20 or SEQ ID NO;

21.

42. The isolated antibody of claim 41, further comprising a light chain variable region comprising an amino acid sequence having at least 95% identity to an amino acid sequence set forth in SEQ ID NO:

22, SEQ ID NO;

24, SEQ ID NO;

25 or SEQ ID NO;

27.

43. An isolated monoclonal antibody, or antigen-binding fragment thereof, that binds to human MASP-2, comprising a light chain variable region comprising any one of the amino acid sequences set forth in SEQ ID NO:

22, SEQ ID NO;

24, SEQ ID NO;

25 or SEQ ID NO;

27.

44. The isolated antibody of claim 43, further comprising a heavy chain variable region comprising an amino acid sequence having at least 95% identity to an amino acid sequence set forth in SEQ ID NO:

18, SEQ ID NO;

20 or SEQ ID NO;

21.

49. An isolated fully human monoclonal antibody or antigen-binding fragment thereof that dissociates from human MASP-2 with a K_D of 10 nM or less as determined by surface plasmon resonance and inhibits C4 activation on a mannan-coated substrate with an IC₅₀ of 10 nM or less in 1% serum.

50. The isolated human antibody of claim 49, wherein said antibody or antigen binding fragment thereof specifically recognizes at least part of an epitope recognized by a reference antibody comprising a heavy chain variable region as set forth in SEQ ID NO:

20 and a light chain variable region as set forth in SEQ ID NO;

24.

56. An article of manufacture comprising a unit dose of a human monoclonal anti-MASP-2 antibody suitable for therapeutic administration to a human subject, wherein the unit dose is the range of from 1 mg to 1000 mg.