Antibody polypeptides that antagonize CD40L
Antibody polypeptides that specifically bind human CD40L are provided. The antibody polypeptides do not activate platelets. The antibody polypeptides are useful in the treatment of diseases involving CD40L activation, such as graft-related diseases and autoimmune diseases. The antibody polypeptides may be domain antibodies (dAbs) comprising a single V H or V K domain. The half-life of the antibody polypeptides may be increased by modifying the antibody polypeptides to be dual specific reagents that can also bind human serum albumin (HSA) or another antigen.
1. An isolated antibody polypeptide comprising a first variable domain that specifically binds human CD40L, wherein CD40L comprises the amino acid sequence of SEQ ID NO: 1, wherein the first variable domain comprises
(a) a CDR1 region having a sequence Trp-X 1 -Leu-Met-Gly (SEQ ID NO: 2), wherein X 1 is Glu or Gln,
(b) a CDR2 region having a sequence Gly-Ile-Glu-Gly-Pro-Gly-Asp-Val-Thr-Tyr-Tyr-Ala-Asp-Ser-Val-Lys-Gly (SEQ ID NO: 3), and
(c) a CDR3 region having a sequence Lys-X 2 -Y 2 -Z 2 -Ser-Asp-Tyr (SEQ ID NO: 4), wherein X 2 is Asp or Glu, Y 2 is Ala or Ser, and Z 2 is Lys, Asn, or Arg.
2. The antibody polypeptide of claim 1 , wherein the first variable domain comprises the amino acid sequence of:
BMS2h-572-10 (SEQ ID NO: 225), BMS2h-572-11 (SEQ ID NO: 226), BMS2h-572-12 (SEQ ID NO: 227), BMS2h-572-14 (SEQ ID NO: 229), BMS2h-572-15 (SEQ ID NO: 230), BMS2h-572-22 (SEQ ID NO: 237), BMS2h-572-23 (SEQ ID NO: 238), BMS2h-572-6 (SEQ ID NO: 243), BMS2h-572-604 (SEQ ID NO: 247), BMS2h-572-607 (SEQ ID NO: 250), BMS2h-572-608 (SEQ ID NO: 251), BMS2h-572-609 (SEQ ID NO: 252), BMS2h-572-610 (SEQ ID NO: 253), BMS2h-572-611 (SEQ ID NO: 254), BMS2h-572-612 (SEQ ID NO: 255), BMS2h-572-613 (SEQ ID NO: 256), BMS2h-572-614 (SEQ ID NO: 257), BMS2h-572-616 (SEQ ID NO: 259), BMS2h-572-619 (SEQ ID NO: 262), BMS2h-572-630 (SEQ ID NO: 271), BMS2h-572-631 (SEQ ID NO: 272), BMS2h-572-632 (SEQ ID NO: 273), BMS2h-572-633 (SEQ ID NO: 274), BMS2h-572-634 (SEQ ID NO: 275), BMS2h-572-635 (SEQ ID NO: 276), BMS2h-572-8 (SEQ ID NO: 278), or BMS2h-572-9 (SEQ ID NO: 279).
3. The antibody polypeptide of claim 1 , wherein the first variable domain comprises the amino acid sequence of BMS2h-572-633 (SEQ ID NO: 274).
4. The antibody polypeptide of claim 1 , wherein the antibody polypeptide is a domain antibody (dAb).
5. The antibody polypeptide of claim 1 , wherein the antibody polypeptide is a fusion polypeptide comprising the first variable domain and an Fc domain.
6. The antibody polypeptide of claim 5 , wherein the fusion polypeptide comprises an IgG4 Fc domain.
7. The antibody polypeptide of claim 5 , wherein the fusion polypeptide comprises an IgG1 Fc domain.
8. The antibody polypeptide of claim 5 , wherein the fusion polypeptide comprises a CT-Long domain (amino acids 139 to 370 of SEQ ID NO: 1362).
9. The antibody polypeptide of claim 5 , wherein the fusion polypeptide comprises a CT-short domain (amino acids 138 to 362 of SEQ ID NO: 1363).
10. The antibody polypeptide of claim 5 , wherein the fusion polypeptide comprises a N297Q Long Fc domain (amino acids 139 to 370 of SEQ ID NO: 1364).
11. The antibody polypeptide of claim 5 , wherein the fusion polypeptide comprises a N297Q Short Fc domain (amino acids 138 to 362 of SEQ ID NO: 1365).
12. The antibody polypeptide of claim 1 , wherein the antibody polypeptide further comprises a second variable domain that specifically binds a second antigen, wherein the second antigen is an antigen other than human CD40L.
13. The antibody polypeptide of claim 12 , wherein the second antigen is a cluster of differentiation (CD) molecule or a Major Histocompatibility Complex (MHC) Class II molecule.
14. The antibody polypeptide of claim 12 , wherein the second antigen is serum albumin (SA).
15. A pharmaceutical composition comprising a therapeutically-effective amount of the antibody polypeptide of claim 1 and a pharmaceutically acceptable carrier.
16. The pharmaceutical composition of claim 15 , further comprising an immunosuppressive/immunomodulatory and/or anti-inflammatory agent.
17. A method of antagonizing CD40L activity in a patient with an immune disease in need of such treatment, comprising administering to the patient a therapeutically effective amount of the pharmaceutical composition of claim 15 .
18. The method of claim 17 , wherein the isolated antibody polypeptide is administered in combination with an immunosuppressive/immunomodulatory and/or anti-inflammatory agent.
19. The method of claim 17 , wherein the immune disease is an autoimmune disease or a graft-related disease.
20. The method of claim 17 , wherein the immune disease is a graft-related disease.
21. The method of claim 20 , wherein the graft-related disease comprises solid organ, tissue and/or cell transplant rejection.
22. The method of claim 20 , wherein the graft-related disease is graft versus host disease (GVHD).
23. The method of claim 20 , wherein the graft-related disease is an acute transplant rejection.
24. The method of claim 20 , wherein the graft-related disease is a chronic transplant rejection.
25. The method of claim 20 , wherein the isolated antibody polypeptide is co-administered with a CTLA4 mutant molecule.
26. The method of claim 25 , wherein the CTLA4 mutant molecule is L104EA29Y-Ig (belatacept).
27. The method of claim 17 , wherein the immune disease is selected from the group consisting of Addison's disease, allergies, ankylosing spondylitis, asthma, atherosclerosis, autoimmune diseases of the ear, autoimmune diseases of the eye, autoimmune hepatitis, autoimmune parotitis, colitis, coronary heart disease, Crohn's disease, diabetes, including Type 1 and/or Type 2 diabetes, epididymitis, glomerulonephritis, Graves' disease, Guillain-Barre syndrome, Hashimoto's disease, hemolytic anemia, idiopathic thrombocytopenic purpura, inflammatory bowel disease, immune response to recombinant drug products, systemic lupus erythematosus, male infertility, multiple sclerosis, myasthenia gravis, pemphigus, psoriasis, rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's syndrome, spondyloarthropathies, thyroiditis, transplant rejection, vasculitis, AIDS, atopic allergy, bronchial asthma, eczema, leprosy, schizophrenia, inherited depression, chronic fatigue syndrome, Alzheimer's disease, Parkinson's disease, myocardial infarction, stroke, autism, epilepsy, Arthus's phenomenon, anaphylaxis, alcohol addiction, and drug addiction.
28. The method of claim 17 , wherein the immune disease is selected from the group consisting of myasthenia gravis, idiopathic thrombocytopenic purpura, and systemic sclerosis.
29. An isolated antibody polypeptide comprising a first variable domain, wherein said antibody polypeptide specifically binds human CD40L, wherein CD40L comprises the amino acid sequence of SEQ ID NO: 1, wherein the antibody polypeptide competes with the binding of BMS2h-572-633, and wherein the antibody polypeptide inhibits binding of CD40L to CD40 with an EC50 of 100 pM to 100 nM.
30. The isolated antibody polypeptide of claim 29 , wherein the first variable domain comprises the amino acid sequence of one of the antibody polypeptides selected from the lineage group consisting of BMS2h-572, BMS2h-719, BMS2h-503, and BMS2h-116.
31. The isolated antibody polypeptide of claim 30 , wherein the first variable domain comprises an amino acid sequence at least 95% identical to BMS2h-572-6, BMS2h-572-608, BMS2h-572-614, BMS2h-572-619, BMS2h-572-633, BMS2h-572-634, BMS2h-572-635, BMS2h-719-2, BMS2h-719-202, BMS2h-719-203, BMS2h-719-213, BMS2h-719-214, BMS2h-719-215, BMS2h-719-218, BMS2h-719-225, BMS2h-503-1, BMS2h-503-2, BMS2h-116-1312, BMS2h-116-1313, or BMS2h-116-1320.
32. The isolated antibody polypeptide of claim 31 , wherein the first variable domain comprises the amino acid sequence of BMS2h-572-6, BMS2h-572-608, BMS2h-572-614, BMS2h-572-619, BMS2h-572-633, BMS2h-572-634, BMS2h-572-635, BMS2h-719-2, BMS2h-719-202, BMS2h-719-203, BMS2h-719-213, BMS2h-719-214, BMS2h-719-215, BMS2h-719-218, BMS2h-719-225, BMS2h-503-1, BMS2h-503-2, BMS2h-116-1312, BMS2h-116-1313, or BMS2h-116-1320.
33. The antibody polypeptide of claim 5 , wherein the amino acid sequence of the fusion polypeptide is set forth in SEQ ID NO: 1355.
34. The antibody polypeptide of claim 5 , wherein the fusion polypeptide comprises the first variable domain consisting of the amino acid sequence of BMS2h-572-633 (SEQ ID NO: 274) and an Fc domain consisting of the amino acid sequence of amino acids 139 to 370 of SEQ ID NO: 1362.
35. The antibody polypeptide of claim 34 , wherein the first variable domain and the Fc domain are linked by an amino acid sequence selected from the group consisting of: AS, AST, TVAAPS (SEQ ID NO: 13), TVA, ASTSGPS (SEQ ID NO: 14) and (GGGGS) n (SEQ ID NO: 12) wherein n is 1, 2, 3, 4, or 5.
36. The pharmaceutical composition of claim 15 , comprising the antibody polypeptide of claim 34 .
37. The pharmaceutical composition of claim 15 , comprising the antibody polypeptide of claim 35 .