Anti-fibril antibodies

Abstract

The invention relates to an isolated monoclonal antibody or antigen binding portion thereof which specifically binds to amyloid fibrils and effectively promotes clearance of extracellular amyloid deposits from the tissues in vivo, as well as humanised antibodies having the same properties.

Description

Abstract This invention relates to an isolated monoclonal antibody or its antigen-binding portion thereof, which specifically binds to amyloid fibrils and effectively promotes the clearance of extracellular amyloid deposits in tissues in vivo, as well as humanized antibodies with the same properties.

Claims

Claims:

1. An isolated monoclonal antibody or antigen binding portion thereof which specifically binds to at least 3 different types of amyloid fibrils, selected from ALK, ALA, ATTR wild type, ATTR variant, AA, AApoAl, ALys, AP2m and AFib amyloid fibrils.

2. An isolated monoclonal antibody or antigen binding portion thereof according to claim 1 , which binds to at least 4, 5, 6, 7, 8 or 9 different types of amyloid fibrils, selected from ALK, ALA, ATTR wild type, ATTR variant, AA, AApoAl, ALys, AP2m and AFib amyloid fibrils.

3. An isolated monoclonal antibody or antigen binding portion thereof which specifically binds to amyloid fibrils and wherein each of CDRH1 , CDRH2 and CDRH3 in the variable domain of the heavy chain has(a) a sequence identity of at least 90% with SEQ ID No 6, SEQ ID No. 7 and SEQ ID NO. 8 respectively; or(b) optionally a single amino acid change with respect to SEQ ID No 6, SEQ ID No. 7 and SEQ ID NO. 8 respectively.

4. An isolated monoclonal antibody or antigen binding portion thereof which specifically binds to amyloid fibrils and wherein each of CDRL1, CDRL2 and CDRL3 in the variable domain of the light chain has(a) a sequence identity of at least 90% with SEQ ID No 9, SEQ ID No. 10 and SEQ ID NO. 11 respectively; or(b) optionally a single amino acid change with respect to SEQ ID No 9, SEQ ID No. 10 and SEQ ID NO. 11 respectively.

5. An isolated monoclonal antibody or antigen binding portion thereof which specifically binds to amyloid fibrils wherein(a) each of CDRH1 , CDRH2 and CDRH3 in the variable domain of the heavy chain has a sequence identity of at least 90% with SEQ ID No 6, SEQ ID No. 7 and SEQ ID NO. 8 respectively and CDRL1, CDRL2 and CDRL3 in the variable domain of the light chain have a sequence identity of at least 90% with SEQ ID No 9, SEQ ID No. 10 and SEQ ID NO. 11 respectively; or(b) each of CDRH1 , CDRH2 and CDRH3 in the variable domain of the heavy chain optionally has a single amino acid change with respect to SEQ ID No 6, SEQ ID No. 7 and SEQ ID NO. 8 respectively, and CDRL1, CDRL2 and CDRL3 in the variable domain of thelight chain optionally have a single amino acid change with respect to SEQ ID No 9, SEQ ID No. 10 and SEQ ID NO. 11 respectively.

6. An isolated monoclonal antibody or antigen binding portion thereof which specifically binds to amyloid fibrils wherein the heavy chain variable region has a sequence identity of at least 70% with SEQ ID No. 2.

7. An isolated monoclonal antibody or antigen binding portion thereof which specifically binds to amyloid fibrils wherein the light chain variable region has a sequence identity of at least 70% with SEQ ID No. 4.

8. An isolated monoclonal antibody or antigen binding portion thereof which specifically binds to amyloid fibrils wherein the heavy chain variable region has a sequence identity of at least 70% with SEQ ID No. 2 and the light chain variable region has a sequence identity of at least 70% with SEQ ID No. 4.

9. An isolated monoclonal antibody or antigen-binding fragment thereof according to any preceding claim, which(a) binds to an epitope present in the amyloid fibril but does not bind to the native peptide from which the fibril is formed; and / or(b) does not bind to Ap fibrils present in the CNS and / or the brain in patients with Alzheimer’s disease; and / or(c) does not compete with SAP for binding to fibrils.

10. An isolated monoclonal antibody or antigen-binding fragment thereof according to claim 9, wherein the epitope is present in at least 3 amyloid fibrils selected from ALK, ALA, ATTR wild type, ATTR variant, AA, AApoAl, ALys, AP2m and AFib amyloid fibrils.

11. An isolated monoclonal antibody or antigen-binding fragment thereof according to claim 9 or claim 10, wherein the epitope comprises the C-terminus of the fibril.

12. An isolated monoclonal antibody or antigen-binding fragment thereof according to any one of claims 9 to 11 , wherein the epitope comprises at least one charged amino acid.

13. An isolated monoclonal antibody or antigen-binding fragment thereof according to any one of claims 9 to 12, wherein the epitope comprises a C-terminal carboxyl group.

14. An isolated monoclonal antibody or antigen-binding fragment thereof according to any one of claims 11 to 13, wherein a citrate ion and / or a malonate ion can be located within the antibody binding site when examined by x-ray crystallography.

15. The isolated monoclonal antibody or antigen binding portion thereof according to any preceding claim wherein the antibody is selected from an IgG, IgA, or an antigen binding antibody fragment selected from an antibody single variable domain polypeptide, dAb, FAb, F(ab’)2, an scFv, an Fv, a VHH domain (such as a Nanobody® or other camelized immunoglobulin domain) or a disulfide-bonded Fv, a human antibody, a chimeric antibody preferably containing a human variable region, a humanized antibody, a bispecific antibody or a single chain antibody.

16. The isolated monoclonal antibody according to any preceding claim wherein the Fc region of the antibody is derived from either the mouse lgG2 or human lgG1 isotype.

17. The isolated monoclonal antibody according to claim 15 or claim 16, which is a humanised antibody.

18. A humanised monoclonal antibody or antigen binding portion thereof according to claim 17, wherein CDRH1 , CDRH2 and CDRH3 in the variable domain of the heavy chain have a sequence identity of at least 90% with SEQ ID No 6, SEQ ID No. 7 and SEQ ID No. 8 respectively.

19. A humanised monoclonal antibody or antigen binding portion thereof according to claim 17 or claim 18, wherein CDRL1, CDRL2 and CDRL3 in the variable domain of the light chain have a sequence identity of at least 90% with SEQ ID No 9, SEQ ID No. 10 and SEQ ID No. 11 respectively.

20. A humanised monoclonal antibody or antigen binding portion thereof according to claim 19 or claim 20, wherein CDRH1 , CDRH2 and CDRH3 in the variable domain of the heavy chain have a sequence identity of at least 90% with SEQ ID No 6, SEQ ID No. 7 and SEQ ID NO. 8 respectively and CDRL1, CDRL2 and CDRL3 in the variable domain of the light chain have a sequence identity of at least 90% with SEQ ID No 9, SEQ ID No. 10 and SEQ ID No. 11 respectively.

21. A humanised monoclonal antibody according to any one of claims 19 to 21 , wherein the human framework regions are derived from antibody genes selected from SEQ ID Nos. 12 and 13.

22. A humanised monoclonal antibody according to any one of claims 18 to 21 , wherein the framework residues mutated to match murine residues include resides selected from VL residues I2, L39, A40, Q44, A49, V101 , N66, T85 and F87, and residues selected from VH residues M39, A80, L55, I66, V25, D85, E69, A45, P46, G47 and K48.

23. A humanised monoclonal antibody according to any one of claims 18 to 22, wherein the VH region comprises the mutations L55K and I66K.

24. A humanised monoclonal antibody according to any one of claims 18 to 23, which has a combination of light and heavy chain framework mutations comprising light chain framework mutations consisting of I2K, A40Y, N66K, T85N and F87Y and heavy chain framework mutations consisting of L55K, I66K, V25A, D85N and E69P.

25. A humanised monoclonal antibody according to any one of claims 1 to 8, which has a combination of light and heavy chain framework mutations comprising light chain framework mutations consisting of I2K, L39M, A40Y, N66K, T85N and F87Y and heavy chain framework mutations consisting of M39I, A80T, L55K, I66K, E69P, A45R, P46T, G47E and K48Q.

26. A humanised monoclonal antibody according to any one of claims 18 to 23, which has a combination of light and heavy chain framework mutations comprising light chain framework mutations consisting of I2K, A40Y, N66K, T85N and F87Y and heavy chain framework mutations consisting of M39I, A80T, L55K, I66K, V25A, D85N and E69P.

27. A humanised monoclonal antibody according to any one of claims 18 to 23 which has a combination of light and heavy chain framework mutations comprising light chain framework mutations consisting of I2K, L39M, A40Y and N66K and heavy chain framework mutations consisting of L55K, I66K, and E69P.

28. A humanised monoclonal antibody according to any one of claims 18 to 24, which has a combination of light and heavy chain framework mutations comprising light chain framework mutations consisting of A40Y and heavy chain framework mutations consisting of L55K, I66K, V25A, D85N and E69P.

29. A humanised monoclonal antibody according to any one of claims 18 to 23, which has a combination of light and heavy chain framework mutations comprising light chain framework mutations consisting of A40Y and heavy chain framework mutations consisting of L55K and I66K.

30. A humanised monoclonal antibody according to claim 17, wherein the VL domain has a sequence identity of at least 90% with SEQ ID No. 14 and the VH domain has a sequence identity of at least 90% with SEQ ID No. 15.

31. A humanised monoclonal antibody according to claim 17, wherein the VL domain has a sequence identity of at least 90% with SEQ ID No. 16 and the VH domain has a sequenceidentity of at least 90% with SEQ ID No. 17.

32. A humanised monoclonal antibody according to claim 17, wherein the VL domain has a sequence identity of at least 90% with SEQ ID No. 18 and the VH domain has a sequence identity of at least 90% with SEQ ID No. 19.

33. A humanised monoclonal antibody according to claim 17, wherein the VL domain has a sequence identity of at least 90% with SEQ ID No. 20 and the VH domain has a sequence identity of at least 90% with SEQ ID No. 21.

34. A humanised monoclonal antibody according to claim 17, wherein the VL domain has a sequence identity of at least 90% with SEQ ID No. 22 and the VH domain has a sequence identity of at least 90% with SEQ ID No. 23.

35. A humanised monoclonal antibody according to claim 17, wherein the VL domain has a sequence identity of at least 90% with SEQ ID No. 24 and the VH domain has a sequence identity of at least 90% with SEQ ID No. 25.

36. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody may have a sequence identity of at least 90% with SEQ ID No. 26 and the VH domain may have a sequence identity of at least 90% with SEQ ID No. 27.

37. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 28 and the VH domain has a sequence identity of at least 90% with SEQ ID No. 29.

38. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 30 and the VH domain has a sequence identity of at least 90% with SEQ ID No. 31.

39. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 32 and the VH domain has a sequence identity of at least 90% with SEQ ID No. 33.

40. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 34 and the VH domain has a sequence identity of at least 90% with SEQ ID No.41 . A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 36 and the VH domain has a sequence identity of at least 90% with SEQ ID No.37.

42. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 38 and the VH domain has a sequence identity of at least 90% with SEQ ID No.39.

43. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 40 and the VH domain has a sequence identity of at least 90% with SEQ ID No.41.

44. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 42 and the VH domain has a sequence identity of at least 90% with SEQ ID No.43.

45. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 44 and the VH domain has a sequence identity of at least 90% with SEQ ID No.45.

46. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 46 and the VH domain has a sequence identity of at least 90% with SEQ ID No.47.

47. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 48 and the VH domain has a sequence identity of at least 90% with SEQ ID No.49.

48. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 50 and the VH domain has a sequence identity of at least 90% with SEQ ID No.51.

49. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 52 and the VH domain has a sequence identity of at least 90% with SEQ ID No.53.

50. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 54 and the VH domain has a sequence identity of at least 90% with SEQ ID No.55.51 . A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 56 and the VH domain has a sequence identity of at least 90% with SEQ ID No.57.

52. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 58 and the VH domain has a sequence identity of at least 90% with SEQ ID No.59.

53. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 60 and the VH domain has a sequence identity of at least 90% with SEQ ID No.61.

54. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 62 and the VH domain has a sequence identity of at least 90% with SEQ ID No.63.

55. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 64 and the VH domain has a sequence identity of at least 90% with SEQ ID No.65.

56. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 66 and the VH domain has a sequence identity of at least 90% with SEQ ID No.67.

57. A humanised monoclonal antibody according to claim 17, wherein the VL domain ofthe isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 68 and the VH domain has a sequence identity of at least 90% with SEQ ID No. 69.

58. A humanised monoclonal antibody according to claim 17, wherein the VL domain of the isolated humanised monoclonal antibody has a sequence identity of at least 90% with SEQ ID No. 70 and the VH domain has a sequence identity of at least 90% with SEQ ID No. 71.

59. A humanised monoclonal antibody or antigen-binding fragment thereof according to any one of claims 18 to 58, which binds to an epitope on amyloid fibrils, wherein the epitope comprises the C-terminus of the fibril.

60. A humanised monoclonal antibody or antigen-binding fragment thereof according to claim 59, wherein the epitope comprises at least one charged amino acid.61 . A humanised monoclonal antibody or antigen-binding fragment thereof according to claim 59 or claim 60, wherein the epitope comprises a C-terminal carboxyl group.

62. A humanised monoclonal antibody or antigen-binding fragment thereof according to any one of claims 59 to 61 , wherein a citrate ion and / or a malonate ion can be located in the antibody binding site when examined by x-ray crystallography.

63. The humanised monoclonal antibody or antigen binding portion thereof according to any one of claims 17 to 62, wherein the antibody is selected from an IgG, IgA, or an antigen binding antibody fragment selected from an antibody single variable domain polypeptide, dAb, FAb, F(ab’)2, an scFv, an Fv, a HH domain (such as a Nanobody® or other camelized immunoglobulin domain) or a disulfide-bonded Fv.

64. The humanised monoclonal antibody according to any one of claims 17 to 63, wherein the Fc region of the antibody is derived from a human lgG1 isotype.

65. The humanised monoclonal antibody or antigen binding portion thereof according to claim 66, wherein the antibody is of hlgG 1 isotype.

66. An isolated monoclonal antibody or a humanised antibody or antigen binding portion thereof according to any preceding claim, which also effectively promotes regression of systemic murine AA amyloid deposits when administered parenterally to mice with experimentally induced systemic AA amyloidosis.

67. An isolated monoclonal antibody or a humanised antibody or antigen binding portion according to any preceding claim, the in vivo efficacy of which is complement dependent.

68. An isolated monoclonal antibody or a humanised antibody or antigen binding portion thereof according to any preceding claim, the in vivo efficacy of which is Fey receptor binding dependent.

69. An isolated monoclonal antibody or a humanised antibody according to any preceding claim, for use in the treatment of disease.

70. An isolated monoclonal antibody or a humanised antibody for use according to claim 22, wherein the disease is amyloidosis.

71. A method for removing amyloid deposits from the tissues in a subject suffering from systemic amyloidosis, comprising administering at least one isolated monoclonal antibody or a humanised antibody according to any one of claims 1 to 70.

72. A pharmaceutical composition comprising an isolated monoclonal antibody or a humanised antibody according to any one of claims 1 to 70 for use in the treatment of systemic amyloidosis.

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