Humanized antibodies that recognize alpha-synuclein
The present application discloses humanized 9E4 antibodies. The antibodies bind to human alpha synuclein and can be used for immunotherapy of Lewy body disease.
1. A method of treating a patient having or at risk of a synucleinopathy, comprising administering to the patient an effective regime of an antibody that binds alpha synuclein, the antibody comprising a humanized heavy chain comprising the three Kabat CDRs of SEQ ID NO:11 and a humanized light chain comprising the three Kabat CDRs of SEQ ID NO:4, provided that position L36 (Kabat numbering) is occupied by F and/or position L83 (Kabat numbering) is occupied by L and/or position H73 (Kabat numbering) is occupied by D, and/or position H93 (Kabat numbering) is occupied by S; wherein a patient at risk of a synucleinopathy has a risk factor conferring a statistically significant risk of developing the synucleinopathy compared with subjects lacking the risk factor.
2. The method of claim 1 , wherein the patient has REM sleep behavior disorder (RBD).
3. A method of detecting Lewy bodies in a patient having or at risk of a Lewy body disease, comprising administering to the patient an effective amount of an antibody that binds alpha synuclein, the antibody comprising a humanized heavy chain comprising the three Kabat CDRs of SEQ ID NO:11 and a humanized light chain comprising the three Kabat CDRs of SEQ ID NO:4, provided that position L36 (Kabat numbering) is occupied by F and/or position L83 (Kabat numbering) is occupied by L and/or position H73 (Kabat numbering) is occupied by D, and/or position H93 (Kabat numbering) is occupied by S; wherein a patient at risk of a synucleinopathy has a risk factor conferring a statistically significant risk of developing the synucleinopathy compared with subjects lacking the risk factor; and detecting bound antibody in the patient.
4. A method of reducing Lewy body formation in a patient having or at risk of a Lewy body disease, comprising administering to the patient an effective amount of an antibody that binds alpha synuclein, the antibody comprising a humanized heavy chain comprising the three Kabat CDRs of SEQ ID NO:11 and a humanized light chain comprising the three Kabat CDRs of SEQ ID NO:4, provided that position L36 (Kabat numbering) is occupied by F and/or position L83 (Kabat numbering) is occupied by L and/or position H73 (Kabat numbering) is occupied by D, and/or position H93 (Kabat numbering) is occupied by S; wherein a patient at risk of a synucleinopathy has a risk factor conferring a statistically significant risk of developing the synucleinopathy compared with subjects lacking the risk factor.
5. A method of inhibiting synuclein aggregation or reducing Lewy bodies or synuclein aggregates in a patient having or at risk of a Lewy body disease, comprising administering to the patient an effective amount of an antibody that binds alpha synuclein, the antibody comprising a humanized heavy chain comprising the three Kabat CDRs of SEQ ID NO:11 and a humanized light chain comprising the three Kabat CDRs of SEQ ID NO:4, provided that position L36 (Kabat numbering) is occupied by F and/or position L83 (Kabat numbering) is occupied by L and/or position H73 (Kabat numbering) is occupied by D, and/or position H93 (Kabat numbering) is occupied by S; wherein a patient at risk of a synucleinopathy has a risk factor conferring a statistically significant risk of developing the synucleinopathy compared with subjects lacking the risk factor.
6. The method of any of claims 1 - 3 , and 4 - 5 , wherein the synucleinopathy or Lewy body disease is Parkinson's disease.
7. The method of claim 1 , wherein the antibody comprises a mature heavy chain variable region having an amino acid sequence designated SEQ ID NO:10 and a mature light chain variable region having an amino acid sequence designated SEQ ID NO:5.
8. The method of claim 1 , wherein position L36 (Kabat numbering) is occupied by F and position L83 (Kabat numbering) is occupied by L.
9. The method of claim 1 , wherein position L36 (Kabat numbering) is occupied by F.
10. The method of claim 1 , wherein position L83 (Kabat numbering) is occupied by L.
11. The method of claim 1 , wherein position H73 (Kabat numbering) is occupied by D.
12. The method of claim 1 , wherein the antibody comprises a mature heavy chain variable region having an amino acid sequence of SEQ ID NO:11 and a mature light chain variable region having an amino acid sequence of SEQ ID NO:4.
13. The method of claim 1 , wherein the mature heavy chain variable region has an amino acid sequence designated SEQ ID NO:10.
14. The method of claim 1 , wherein the mature light chain variable region has an amino acid sequence designated SEQ ID NO: 5.
15. The method of claim 1 , wherein position H73 (Kabat numbering) is occupied by D and position H93 (Kabat numbering) is occupied by A.
16. The method of claim 1 , wherein the mature light chain variable region has an amino acid sequence of SEQ ID NO:3, 4, or 5, and the mature heavy chain variable region has an amino acid sequence of SEQ ID NO:8, 9, or 10.
17. The method of claim 16 , wherein the antibody further comprises a heavy chain constant region having the amino acid sequence of SEQ ID NO:32.
18. The method of claim 16 , wherein the antibody further comprises a light chain constant region having the amino acid sequence of SEQ ID NO:13.
19. The method of claim 1 , wherein the antibody further comprises a heavy chain constant region of human IgG1 isotype.
20. The method of claim 1 , wherein the antibody is a Fab fragment.
21. The method of claim 3 , wherein position L36 (Kabat numbering) is occupied by F and position L83 (Kabat numbering) is occupied by L.
22. The method of claim 3 , wherein position L36 (Kabat numbering) is occupied by F.
23. The method of claim 3 , wherein position L83 (Kabat numbering) is occupied by L.
24. The method of claim 3 , wherein position H73 (Kabat numbering) is occupied by D.
25. The method of claim 3 , wherein the antibody comprises a mature heavy chain variable region having an amino acid sequence of SEQ ID NO:11 and a mature light chain variable region having an amino acid sequence of SEQ ID NO:4.
26. The method of claim 3 , wherein the mature heavy chain variable region has an amino acid sequence designated SEQ ID NO:10 and the mature light chain variable region has an amino acid sequence designated SEQ ID NO: 5.
27. The method of claim 3 , wherein position H73 (Kabat numbering) is occupied by D and position H93 (Kabat numbering) is occupied by A.
28. The method of claim 3 , wherein the mature light chain variable region has an amino acid sequence of SEQ ID NO:3, 4, or 5, and the mature heavy chain variable region has an amino acid sequence of SEQ ID NO:8, 9, or 10.
29. The method of claim 28 , wherein the antibody further comprises a heavy chain constant region having the amino acid sequence of SEQ ID NO:32.
30. The method of claim 28 , wherein the antibody further comprises a light chain constant region having the amino acid sequence of SEQ ID NO:13.
31. The method of claim 3 , wherein the antibody further comprises a heavy chain constant region of human IgG1 isotype.
32. The method of claim 3 , wherein the antibody is a Fab fragment.
33. The method of claim 4 , wherein the antibody comprises a mature heavy chain variable region having an amino acid sequence designated SEQ ID NO:10 and a mature light chain variable region having an amino acid sequence designated SEQ ID NO:5.
34. The method of claim 4 , wherein position L36 (Kabat numbering) is occupied by F and position L83 (Kabat numbering) is occupied by L.
35. The method of claim 4 , wherein position L36 (Kabat numbering) is occupied by F.
36. The method of claim 4 , wherein position L83 (Kabat numbering) is occupied by L.
37. The method of claim 4 , wherein position H73 (Kabat numbering) is occupied by D.
38. The method of claim 4 , wherein the antibody comprises a mature heavy chain variable region having an amino acid sequence of SEQ ID NO:11 and a mature light chain variable region having an amino acid sequence of SEQ ID NO:4.
39. The method of claim 4 , wherein the mature heavy chain variable region has an amino acid sequence designated SEQ ID NO:10 and the mature light chain variable region has an amino acid sequence designated SEQ ID NO: 5.
40. The method of claim 4 , wherein position H73 (Kabat numbering) is occupied by D and position H93 (Kabat numbering) is occupied by A.
41. The method of claim 4 , wherein the mature light chain variable region has an amino acid sequence of SEQ ID NO:3, 4, or 5, and the mature heavy chain variable region has an amino acid sequence of SEQ ID NO:8, 9, or 10.
42. The method of claim 41 , wherein the antibody further comprises a heavy chain constant region having the amino acid sequence of SEQ ID NO:32.
43. The method of claim 41 , wherein the antibody further comprises a light chain constant region having the amino acid sequence of SEQ ID NO:13.
44. The method of claim 4 , wherein the antibody further comprises a heavy chain constant region of human IgG1 isotype.
45. The method of claim 4 , wherein the antibody is a Fab fragment.
46. The method of claim 5 , wherein the antibody further comprises a mature heavy chain variable region having an amino acid sequence designated SEQ ID NO:10 and a mature light chain variable region having an amino acid sequence designated SEQ ID NO:5.
47. The method of claim 5 , wherein position L36 (Kabat numbering) is occupied by F and position L83 (Kabat numbering) is occupied by L.
48. The method of claim 5 , wherein position L36 (Kabat numbering) is occupied by F.
49. The method of claim 5 , wherein position L83 (Kabat numbering) is occupied by L.
50. The method of claim 5 , wherein position H73 (Kabat numbering) is occupied by D.
51. The method of claim 5 , wherein the antibody further comprises a mature heavy chain variable region having an amino acid sequence of SEQ ID NO:11 and a mature light chain variable region having an amino acid sequence of SEQ ID NO:4.
52. The method of claim 5 , wherein the mature heavy chain variable region has an amino acid sequence designated SEQ ID NO:10.
53. The method of claim 5 , wherein the mature light chain variable region has an amino acid sequence designated SEQ ID NO: 5.
54. The method of claim 5 , wherein position H73 (Kabat numbering) is occupied by D and position H93 (Kabat numbering) is occupied by A.
55. The method of claim 5 , wherein the mature light chain variable region has an amino acid sequence of SEQ ID NO:3, 4, or 5, and the mature heavy chain variable region has an amino acid sequence of SEQ ID NO:8, 9, or 10.
56. The method of claim 5 , wherein the antibody further comprises a heavy chain constant region having the amino acid sequence of SEQ ID NO:32.
57. The method of claim 55 , wherein the antibody further comprises a light chain constant region having the amino acid sequence of SEQ ID NO:13.
58. The method of claim 5 , wherein the antibody further comprises a heavy chain constant region of human IgG1 isotype.
59. The method of claim 5 , wherein the antibody is a Fab fragment.