Antibodies to TNF-α and use thereof
View Patent ↗The present invention is directed to antibodies and fragments thereof having binding specificity for TNF-α. Another embodiment of this invention relates to the antibodies described herein, and binding fragments thereof, comprising the sequences of the V H , V L and CDR polypeptides described herein, and the polynucleotides encoding them. The invention also contemplates conjugates of anti-TNF-α antibodies and binding fragments thereof conjugated to one or more functional or detectable moieties. The invention also contemplates methods of making said anti-TNF-α antibodies and binding fragments thereof. Embodiments of the invention also pertain to the use of anti-TNF-α antibodies, and binding fragments thereof, for the diagnosis, assessment and treatment of diseases and disorders associated with TNF-α.
1. An anti-human TNF-α antibody or antigen-binding fragment thereof which specifically binds to human TNF-α, wherein said antibody or antigen-binding fragment comprises the V H CDR polypeptide sequences of SEQ ID NO: 7, 8, and 9 and the V L CDR polypeptide sequences of SEQ ID NO: 4, 5, and 6.
2. The antibody or antigen-binding fragment thereof of claim 1 , wherein said antibody or fragment is aglycosylated.
3. The antibody of claim 1 , wherein said antibody or antigen-binding fragment comprises an Fc region that has been modified to alter effector function, half-life, proteolysis, and/or glycosylation.
4. The antibody of claim 1 , wherein said antibody or antigen-binding fragment is a human, humanized, single chain, or chimeric antibody.
5. The antibody of claim 4 , wherein said antibody or fragment is a humanized antibody derived from a rabbit (parent) anti-human TNF-α antibody.
6. antibody or antigen-binding fragment of claim 1 , wherein said antibody or antigen-binding fragment is directly or indirectly attached to a detectable label or therapeutic agent.
7. The antibody of claim 1 , wherein said antibody or antigen-binding fragment comprises the V H polypeptide sequence of SEQ ID NO: 3 and the V L polypeptide of SEQ ID NO: 2 wherein one or more of the framework residues (FR residues) in said V H or V L polypeptides have been substituted with another amino acid residue resulting in an anti-TNF-α antibody that retains specific binding to human TNF-α.
8. The antibody of claim 7 , wherein said antibody is humanized.
9. The antibody of claim 7 , wherein said antibody is chimeric.
10. The antibody of claim 9 , wherein said chimeric antibody comprises a human F c .
11. The antibody of claim 10 , wherein said human F c is derived from IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, IgG7, IgG8, IgG9, IgG10, IgG11, IgG12, IgG13, IgG14, IgG15, IgG16, IgG17, IgG18 or IgG19.
12. The antibody of claim 7 , wherein said antibody inhibits the association of TNF-α with TNF-R and/or multimers thereof and/or antagonizes the biological effects thereof.
13. The antibody of claim 12 , wherein said antibody binds to TNF-α with a dissociation constant (K D ) of less than or equal to 5×10 −7 , 10 −7 , 5×10 −8 , 10 −8 , 5×10 −9 , 10 −9 , 5×10 −10 , 10 −10 , 5×10 −11 , 10 −11 , 5×10 −2 , 10 −12 , 5×10 13 , or 10 −13 .
14. The antibody of claim 13 , wherein said antibody binds to TNF-α with a dissociation constant (K D ) of less than or equal to 5×10 −10 .
15. The antibody of claim 12 , wherein said antibody binds to TNF-αc with an off-rate (K off ) of less than or equal to 10 −4 S −1 , 5×10 −5 S −1 , 10 −5 S −1 , 5×10 −6 S −1 , 10 −6 S −1 , 5×10 −7 S −1 , or 10 −7 S −1 .
16. The antibody of claim 7 , wherein said antibody inhibits the production of TNF-α/TNF-R complexes and/or the production of TNF-α/TNF-R multimers.
17. The antibody of claim 16 , wherein the TNF-R is p75 TNF-R.
18. The antibody of claim 7 , wherein said fragment is selected from an F ab fragment, an F ab′ fragment, or an F (ab′)2 fragment.
19. The antibody of claim 7 , wherein said antibody further comprises an effector moiety.
20. The antibody of claim 19 , wherein the effector moiety is a detectable moiety or a functional moiety.
21. The antibody of claim 20 , wherein said detectable moiety is a fluorescent dye, an enzyme, a substrate, a bioluminescent material, a radioactive material, or a chemiluminescent material.
22. The antibody of claim 20 , wherein said functional moiety is streptavidin, avidin, biotin, a cytotoxin, a cytotoxic agent, or a radioactive material.
23. An anti-TNF-α antibody or antigen-binding fragment thereof according to claim 1 that binds human TNF-α, wherein said antibody or antigen-binding fragment comprises V H and V L polypeptides which each are at least 90% identical to the V H chain of the polypeptide of SEQ ID NO: 3 and the V L chain of the polypeptide sequence of SEQ ID NO: 2.
24. An isolated anti-TNF-α antibody or antibody fragment thereof according to claim 1 , comprising the V H polypeptide sequence of SEQ ID NO: 3 or the V L polypeptide sequence of SEQ ID NO: 2.
25. The antibody of claim 2 , wherein said antibody is humanized.
26. The antibody of claim 2 , wherein said antibody is chimeric.
27. The antibody of claim 26 , wherein said chimeric antibody comprises a human F c .
28. The antibody of claim 1 , which comprises a single chain antibody.
29. The antibody of claim 27 , wherein said human F c is derived from human IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, IgG7, IgG8, IgG9, IgG10, IgG11, IgG12, IgG13, IgG14, IgG15, IgG16, IgG17, IgG18 or IgG19.
30. The anti-TNF-α antibody of claim 23 , wherein said antibody comprises a V L polypeptide sequence having at least 95% or greater homology to SEQ ID NO: 2 or a V H polypeptide sequence having at least 95% or greater homology to SEQ ID NO: 3.
31. The antibody of claim 23 , wherein said antibody binds to TNF-α with a dissociation constant (K D ) of less than or equal to 5×10 −7 , 10 −7 , 5×10 −8 , 10 −8 , 5×10 −9 , 10 −9 , 5×10 −10 , 10 −10 , 5×10 −11 , 10 −11 , 5×10 −12 , 10 −12 , 5×10 13 , or 10 −13 .
32. The antibody of claim 23 , wherein said antibody binds to TNF-α with a dissociation constant (K D ) of less than or equal to 5×10 −10 .
33. The antibody of claim 23 , wherein said antibody binds to TNF-α with an off-rate (k off ) of less than or equal to 10 −4 S −1 , 5×10 −5 S − 1, 10 −5 S −1 , 5×10 −6 S −1 , 10 −6 S −1 , 5×10 −7 S −1 , or 10 −7 S −1 .
34. The antibody of claim 23 , wherein said antibody inhibits the association of TNF-α with TNF-R and/or multimers thereof and/or antagonizes the biological effects thereof.
35. The antibody of claim 34 , wherein the TNF-R is p55 TNF-R or p75 TNF-R.
36. The antibody of claim 23 , wherein said fragment is selected from an F ab fragment, an F ab′ fragment, or an F (ab′)2 fragment.
37. The antibody of claim 23 , wherein said antibody further comprises an effector moiety.
38. The antibody of claim 37 , wherein the effector moiety is a detectable moiety or a functional moiety.
39. The antibody of claim 38 , wherein said detectable moiety is a fluorescent dye, an enzyme, a substrate, a bioluminescent material, a radioactive material, or a chemiluminescent material.
40. The antibody of claim 38 , wherein said functional moiety is streptavidin, avidin, biotin, a cytotoxin, a cytotoxic agent, or a radioactive material.
41. A pharmaceutical or diagnostic composition comprising the TNF-α antibody or antigen-binding fragment of claim 1 and a pharmaceutically acceptable carrier.
42. The pharmaceutical or diagnostic composition of claim 41 , further comprising at least one stabilizer.
43. The pharmaceutical or diagnostic composition of claim 41 , wherein said composition is lyophilized.
44. The antibody or antigen-binding fragment thereof of claim 1 , wherein said antibody is a humanized or chimeric antibody or antibody fragment.
45. A composition comprising the antibody or antigen-binding fragment of claim 44 and a pharmaceutically or diagnostically acceptable carrier or excipient.
46. The antibody of claim 23 , wherein said V L polypeptide comprises the CDR polypeptide sequences of SEQ ID NO: 4, 5, or 6; and wherein said V H polypeptide comprises the CDR polypeptide sequences of SEQ ID NO: 7, 8, or 9.