Humanized antibodies against monocyte chemotactric proteins
View Patent ↗The invention provides humanized antibodies that bind to a plurality of b-chemokines, particularly monocyte chemotactic proteins MCP-1, MCP-2 and MCP-3. The invention also provides therapeutic reagents and methods of treating disorders associated with detrimental MCP activity.
1. A humanized immunoglobulin or antigen-binding fragment thereof comprising a) heavy chain complementarity determining regions as set forth in SEQ ID NO:29, SEQ ID NO:30, and SEQ ID NO:31, and b) light chain complementarity determining regions as set forth in SEQ ID NO:32, SEQ ID NO:33, and SEQ ID NO:34.
2. The humanized immunoglobulin or antigen-binding fragment of claim 1 , wherein the heavy chain further comprises variable region framework residues L27, I29, and T73 (Kabat numbering convention) from the monoclonal antibody 11K2 heavy chain as set forth in SEQ ID NO:27.
3. The humanized immunoglobulin or antigen-binding fragment of claim 1 , wherein the heavy chain further comprises least one variable region framework residue selected from the group consisting of N28, K30, I48, and A67 (Kabat numbering convention) from the monoclonal antibody 11K2 heavy chain as set forth in SEQ ID NO:27.
4. The humanized immunoglobulin or antigen-binding fragment of claim 1 , wherein the heavy chain further comprises variable region framework residues N28, K30, I48, and A67 (Kabat numbering convention) from the monoclonal antibody 11K2 heavy chain as set forth in SEQ ID NO:27.
5. The humanized immunoglobulin or antigen-binding fragment of claim 1 , wherein the light chain further comprises at least one variable region framework residue from the monoclonal antibody 11K2 light chain set forth as SEQ ID NO: 28, wherein the residue is selected from the group consisting of S49 and Y71 (Kabat numbering convention), and wherein the remainder of the light chain is from a human immunoglobulin.
6. The humanized immunoglobulin or antigen-binding fragment of claim 1 , wherein the light chain further comprises variable region framework residues S49 and Y71 (Kabat numbering convention) from the monoclonal antibody 11K2 light chain as set forth in SEQ ID NO:28.
7. The humanized immunoglobulin or antigen-binding fragment of claim 6 , wherein the light chain further comprises variable region framework residue K69 (Kabat numbering convention) from the monoclonal antibody 11K2 light chain as set forth in SEQ ID NO:28.
8. The humanized immunoglobulin or antigen-binding fragment of claim 1 , wherein the heavy chain further comprises at least one variable region framework residue from the monoclonal antibody 11K2 heavy chain set forth as SEQ ID NO: 27, wherein the residue is selected from the group consisting of L27, N28, I29, K30, I48, A67, and T73 (Kabat numbering).
9. The humanized immunoglobulin or antigen-binding fragment of claim 8 , wherein the heavy chain comprises variable region framework residues L27, N28, I29, K30, I48, A67, and T73 (Kabat numbering convention) from the monoclonal antibody 11K2 heavy chain as set forth in SEQ ID NO:27.
10. The humanized immunoglobulin or antigen-binding fragment of claim 6 , wherein the heavy chain comprises at least one variable region framework residues L27, I29, and T73 (Kabat numbering convention) from the monoclonal antibody 11K2 heavy chain as set forth in SEQ ID NO:27.
11. The humanized immunoglobulin or antigen-binding fragment of claim 9 , wherein the light chain comprises at least one variable region framework residue from the monoclonal antibody 11K2 light chain set forth as SEQ ID NO: 28, wherein the residue is selected from the group consisting of S49, K69, and Y71 (Kabat numbering).
12. The humanized immunoglobulin or antigen-binding fragment of claim 1 , wherein the heavy chain comprises at least one variable region framework residue from the monoclonal antibody 11K2 heavy chain set forth as SEQ ID NO: 27, wherein the residue is selected from the group consisting of L27, N28, I29, K30, I48, A67 and T73 (Kabat numbering convention), and the light chain comprises at least one variable region framework residue from the monoclonal antibody 11K2 light chain set forth as SEQ ID NO: 28, wherein the residue is selected from the group consisting of S49, K69, and Y71 (Kabat numbering convention), wherein the remainder of the heavy and light chains are from a human immunoglobulin.
13. The humanized immunoglobulin or antigen-binding fragment of claim 12 , wherein the heavy chain comprises variable region framework residues L27, I29, and T73, and the light chain comprises variable region framework residues S49 and Y71.
14. The humanized immunoglobulin or antigen-binding fragment of claim 12 , wherein the heavy chain comprises variable region framework residues L27, I29, and T73, and the light chain comprises variable region framework residues S49, K69, and Y71.
15. The humanized immunoglobulin or antigen-binding fragment of claim 12 , wherein the heavy chain comprises variable region framework residues L27, N28, I29, K30, I48, A67 and T73, and the light chain comprises variable region framework residues S49 and Y71.
16. A humanized immunoglobulin or antigen-binding fragment comprising a) heavy chain complementary determining regions as set forth in SEQ ID NO: 29, SEQ ID NO: 30, and SEQ ID NO: 31, and variable region framework residues L27, N28, I29, K30, I48, A67, and T73 (Kabat numbering) from the monoclonal antibody 11K2 heavy chain set forth as SEQ ID NO: 27, and b) light chain complementary determining regions as set forth in SEQ ID NO: 32, SEQ ID NO: 33, and SEQ ID NO: 34, and variable region framework residues S49, K69, and Y71 (Kabat numbering) from the monoclonal antibody 11K2 light chain set forth as SEQ ID NO: 28, wherein the remainder of the heavy and light chains are from a human immunoglobulin.
17. The immunoglobulin or antigen-binding fragment of claim 16 , wherein the immunoglobulin is modified by reducing or eliminating at least one potential glycosylation site.
18. The immunoglobulin or antigen-binding fragment of claim 1 , which binds to MCP-1.
19. The immunoglobulin or antigen-binding fragment of claim 1 , which specifically binds to MCP-1 with a binding affinity of at least 10 −9 M.
20. The immunoglobulin or antigen-binding fragment of claim 1 , which specifically binds to MCP-1 with a binding affinity of at least 10 −10 M.
21. The immunoglobulin or antigen-binding fragment of claim 1 , which specifically binds to MCP-1 with a binding affinity of at least 10 −11 M.
22. The immunoglobulin or antigen-binding fragment of claim 1 , which further binds to MCP-2 with a binding affinity of at least 10 −7 M.
23. The immunoglobulin or antigen-binding fragment of claim 1 , which further binds to MCP-2 with a binding affinity of at least 10 −8 M.
24. The immunoglobulin or antigen-binding fragment of claim 1 , which further binds to MCP-2 with a binding affinity of at least 10 −9 M.
25. The immunoglobulin or antigen-binding fragment of claim 1 , which binds to MCP-2.
26. The immunoglobulin or antigen-binding fragment of claim 1 , which specifically binds to MCP-2 with a binding affinity of at least 10 −7 M.
27. The immunoglobulin or antigen-binding fragment of claim 1 , which specifically binds to MCP-2 with a binding affinity of at least 10 −8 M.
28. The immunoglobulin or antigen-binding fragment of claim 1 , which specifically binds to MCP-2 with a binding affinity of at least 10 −9 M.
29. The immunoglobulin or antigen-binding fragment of claim 1 , which binds to MCP-1 and MCP-2.
30. The immunoglobulin or antigen-binding fragment of claim 1 , which binds to an epitope within MCP-1, MCP-2, and MCP-3.
31. An antibody comprising the same heavy and light chain polypeptide sequences as an antibody produced by clone 3F2 (ATCC patent deposit designation PTA-5308).
32. An immunoglobulin or antigen-binding fragment of claim 1 , wherein the heavy chain isotype is gamma 1.
33. An immunoglobulin or antigen-binding fragment of claim 1 , wherein the antibody is modified by reducing or eliminating at least one potential glycosylation site.
34. An immunoglobulin or antigen-binding fragment of claim 1 , wherein the antibody is modified by conjugation to a carrier selected from polyethylene glycol and albumen.
35. An antibody or antigen-binding fragment of claim 1 , wherein the constant region of the antibody is modified to reduce at least one constant region-mediated biological effector function relative to an unmodified antibody.
36. A fragment of claim 1 , wherein the fragment is a Fab fragment.
37. An immunoglobulin or antigen-binding fragment of claim 1 , which inhibits MCP-induced chemotaxis.
38. An immunoglobulin or antigen-binding fragment of claim 37 , wherein the immunoglobulin or antigen-binding fragment inhibits MCP-1-induced chemotaxis, MCP-2-induced chemotaxis, or both MCP-1-induced and MCP-2-induced chemotaxis.
39. An immunoglobulin or antigen-binding fragment of claim 1 , which inhibits MCP-induced collagen expression.
40. An immunoglobulin or antigen-binding fragment of claim 39 , wherein the immunoglobulin or antigen-binding fragment inhibits MCP-1-induced collagen expression, MCP-2-induced collagen expression, or both MCP-1-induced and MCP-2-induced collagen expression.
41. An immunoglobulin or antigen-binding fragment of claim 1 , which inhibits MCP-1 induced angiogenesis.
42. An immunoglobulin or antigen-binding fragment of claim 41 , wherein the immunoglobulin or antigen-binding fragment inhibits MCP-1-induced angiogenesis, MCP-2-induced angiogenesis, or both MCP-1-induced and MCP-2-induced angiogenesis.
43. An immunoglobulin or antigen-binding fragment of claim 1 , which reduces inflammation in a subject.
44. The immunoglobulin or antigen-binding fragment of claim 43 , wherein the inflammation is associated with a disorder selected from the group consisting of arthritis, multiple sclerosis, cirrhosis, atherosclerosis, and breast carcinoma.
45. An immunoglobulin or antigen-binding fragment of claim 1 , which reduces fibrosis in a subject.
46. A pharmaceutical composition comprising the immunoglobulin or antigen-binding fragment of claim 1 and a pharmaceutical carrier.