Humanized tau antibodies in a alzheimer's disease
The present invention is in the fields of biochemistry, molecular biology, and Alzheimer's disease diagnosis, prevention, and treatment Provided herein are humanized antibodies against human tau that are capable of discriminating between normal (healthy) and pathological (disease-associated) tau.
1. A humanized anti-tau antibody, or a tau-binding fragment thereof, wherein said antibody or binding fragment comprises:
a heavy chain variable region comprising CDR-H1, CDR-H2, and CDR-H3 of SEQ ID NOs. 1, 2, and 3, respectively, and a framework from human immunoglobulin M65092 (SEQ ID NO. 71);
a light chain variable region comprising CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NOs. 4, 5, and 6, respectively, and a framework from human immunoglobulin X72449 (SEQ ID NO. 65); and
heavy chain and light chain constant regions each from a human immunoglobulin; and wherein said heavy chain framework has been substituted at one or more positions selected from 9, 21, 27, 28, 30, 38, 48, 67, 68, 70, and 95; said light chain framework has either not been substituted or has been substituted at position 5; and wherein said positions are according to Kabat.
2. The antibody or binding fragment of claim 1 , wherein heavy chain position 9 is occupied by P, position 21 is occupied by P, position 27 is occupied by Y, position 28 is occupied by I, position 30 is occupied by T, position 38 is occupied by K, position 48 is occupied by I, position 67 is occupied by K, position 68 is occupied by A, position 70 is occupied by L, and/or position 95 is occupied by F.
3. The antibody or binding fragment of claim 1 , wherein light chain position 5 is occupied by S.
4. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region is selected from SEQ ID NOs. 13-25 (RHA through RHM); and the sequence of the light chain variable region is SEQ ID NO. 26 (RKA).
5. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region is selected from SEQ ID NOs. 13-25 (RHA through RHM); and the sequence of the light chain variable region is SEQ ID NO. 27 (RKB).
6. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 14 and the sequence of the light chain variable region comprises SEQ ID NO. 26.
7. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 15 and the sequence of the light chain variable region comprises SEQ ID NO. 26.
8. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 16 and the sequence of the light chain variable region comprises SEQ ID NO. 26.
9. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 17 and the sequence of the light chain variable region comprises SEQ ID NO. 26.
10. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 18 and the sequence of the light chain variable region comprises SEQ ID NO. 26.
11. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 19 and the sequence of the light chain variable region comprises SEQ ID NO. 26.
12. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 20 and the sequence of the light chain variable region comprises SEQ ID NO. 26.
13. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 21 and the sequence of the light chain variable region comprises SEQ ID NO. 26.
14. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 22 and the sequence of the light chain variable region comprises SEQ ID NO. 26.
15. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 23 and the sequence of the light chain variable region comprises SEQ ID NO. 26.
16. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 24 and the sequence of the light chain variable region comprises SEQ ID NO. 26.
17. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 25 and the sequence of the light chain variable region comprises SEQ ID NO. 26.
18. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 13 and the sequence of the light chain variable region comprises SEQ ID NO. 27.
19. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 14 and the sequence of the light chain variable region comprises SEQ ID NO. 27.
20. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 15 and the sequence of the light chain variable region comprises SEQ ID NO. 27.
21. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 16 and the sequence of the light chain variable region comprises SEQ ID NO. 27.
22. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 17 and the sequence of the light chain variable region comprises SEQ ID NO. 27.
23. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 18 and the sequence of the light chain variable region comprises SEQ ID NO. 27.
24. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 19 and the sequence of the light chain variable region comprises SEQ ID NO. 27.
25. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 20 and the sequence of the light chain variable region comprises SEQ ID NO. 27.
26. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 21 and the sequence of the light chain variable region comprises SEQ ID NO. 27.
27. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 22 and the sequence of the light chain variable region comprises SEQ ID NO. 27.
28. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 23 and the sequence of the light chain variable region comprises SEQ ID NO. 27.
29. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 24 and the sequence of the light chain variable region comprises SEQ ID NO. 27.
30. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 25 and the sequence of the light chain variable region comprises SEQ ID NO. 27.
31. The antibody or binding fragment of claim 1 wherein the antibody is of the IgG1 isotype.
32. The antibody or binding fragment of claim 1 wherein the antibody is of the IgG4 isotype.
33. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 14, the sequence of the light chain variable region comprises SEQ ID NO. 26, and the antibody is of the IgG1 isotype.
34. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 16, the sequence of the light chain variable region comprises SEQ ID NO. 26, and the antibody is of the IgG1 isotype.
35. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 17, the sequence of the light chain variable region comprises SEQ ID NO. 26, and the antibody is of the IgG1 isotype.
36. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 25, the sequence of the light chain variable region comprises SEQ ID NO. 26, and the antibody is of the IgG1 isotype.
37. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 16, the sequence of the light chain variable region comprises SEQ ID NO. 27, and the antibody is of the IgG1 isotype.
38. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 17, the sequence of the light chain variable region comprises SEQ ID NO. 27, and the antibody is of the IgG1 isotype.
39. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 16, the sequence of the light chain variable region comprises SEQ ID NO. 27, and the antibody is of the IgG4 isotype.
40. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region is comprises ID NO. 17, the sequence of the light chain variable region comprises SEQ ID NO. 27, and the antibody is of the IgG4 isotype.
41. An antibody, or a tau-binding fragment thereof, according to claim 1 wherein said antibody or binding fragment binds at least one epitope selected from the group consisting of HQPGGG (SEQ ID NO: 148), HVPGGG (SEQ ID NO: 149), and HKPGGG (SEQ ID NO: 150).
42. An immunoconjugate having the formula (A)-(L)-(C), wherein: (A) is an antibody or binding fragment thereof of claim 1 ; (L) is a linker; and (C) is an agent; and wherein said linker (L) links (A) to (C).
43. A nucleic acid molecule encoding any of the antibodies or binding fragments of claim 1 .
44. A vector comprising a nucleic acid according to claim 43 .
45. A host cell comprising a vector according to claim 44 .
46. A method of producing an antibody or tau-binding fragment thereof that binds to human tau comprising culturing the host cell of claim 45 so that the nucleic acid is expressed and the antibody, tau-binding fragment, heavy chain, or light chain thereof produced.
47. The antibody or binding fragment of claim 1 , wherein the sequence of the heavy chain variable region comprises SEQ ID NO. 13 and the sequence of the light chain variable region comprises SEQ ID NO. 26.
48. The antibody or binding fragment of claim 47 wherein the antibody is of the IgG1 isotype.
49. The antibody or binding fragment of claim 47 wherein the antibody is of the IgG4 isotype.
50. A pharmaceutical composition comprising the antibody or binding fragment according to claim 1 and a pharmaceutically acceptable carrier, diluent, excipient, or stabilizer.
51. A method of treating Alzheimer's Disease or another tauopathy in a subject having Alzheimer's Disease or another tauopathy, comprising administering to the subject a therapeutically effective amount of (1) a composition comprising an antibody or tau-binding fragment according to claim 1 , or (2) a composition according to claim 50 .
52. A method of promoting clearance of tau aggregates from the brain of a subject having, suspected of having, or being prone to have Alzheimer's Disease or another tauopathy, comprising administering to the subject a therapeutically effective amount of (1) an antibody or tau-binding fragment according to claim 1 , or (2) a composition according to claim 50 .
53. A method of slowing progression of AD or another tauopathy in a subject having Alzheimer's Disease or another tauopathy, comprising administering to the subject a therapeutically effective amount of (1) an antibody or tau-binding fragment according to claim 1 , or (2) a composition according to claim 50 .
54. A method of ameliorating the symptoms of AD or another tauopathy in a subject having Alzheimer's Disease or another tauopathy, comprising administering to the subject a therapeutically effective amount of (1) an antibody or tau-binding fragment according to claim 1 , or (2) a composition according to claim 50 .
55. A method of treating cognitive impairment in a subject having Alzheimer's Disease or another tauopathy, comprising administering to the subject a therapeutically effective amount of (1) an antibody or tau-binding fragment according to claim 1 , or (2) a composition according to claim 50 .
56. A humanized anti-tau antibody, or a tau-binding fragment thereof, wherein said antibody or binding fragment comprises:
a. a heavy chain variable region comprising CDR-H1, CDR-H2, and CDR-H3 of SEQ ID NOs. 1, 2, and 3, respectively, and a framework from human immunoglobulin M65092 (SEQ ID NO. 71); and
b. a light chain variable region comprising CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NOs. 4, 5, and 6, respectively, and a framework from human immunoglobulin X72449 (SEQ ID NO. 65); and
c. heavy chain and light chain constant regions from a human immunoglobulin.
57. An anti-tau antibody, or a tau-binding fragment thereof, wherein said antibody or binding fragment comprises:
a. a heavy chain having the amino acid sequence of any one of SEQ ID NO. 28-40 and 43-55; and
b. a light chain having the amino acid sequence of any one of SEQ ID NO. 57 and 58.
58. A method of evaluating a subject having, suspected of having, or being prone to have Alzheimer's Disease or another tauopathy, the method comprising the step of detecting binding of an antibody or tau-binding fragment of claim 1 to a component of a biological sample from the subject, wherein the detection of binding to the biological sample is indicative of Alzheimer's Disease or another tauopathy in the subject.