AU2013203262B2

AU2013203262B2 - Humanized anti-CXCR5 antibodies, derivatives thereof and their uses

Info

Publication number: AU2013203262B2
Application number: AU2013203262A
Authority: AU
Inventors: Elizabeth Allen; Nicolas Baurin; Beatrice Cameron; Renata Lee; Vincent Mikol; Thomas Oligino; Norman Ruetsch
Original assignee: Sanofi SA
Current assignee: Sanofi SA
Priority date: 2007-08-29
Filing date: 2013-04-09
Publication date: 2016-07-07
Anticipated expiration: 2028-08-27
Also published as: AU2013203261B2; AU2013203265A1; AU2013203264B2; AU2013203261A1; AU2013203262A1; AU2013203264A1; AU2013203265B2

Abstract

The present invention relates to humanized antibodies that specifically bind to CXCR5 and can, for example, inhibit CXCR5 function. The invention also includes uses of the antibodies to treat or prevent CXCR5 related diseases or disorders.

Description

THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS: 1. An isolated antibody or fragment thereof that competitively inhibits the binding of an anti-CXCR5 antibody or fragment thereof to the extracellular domain of human CXCR5, wherein the anti-CXCR5 antibody or fragment thereof comprises: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 11, and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 12; (b) the amino acid sequences of RSSKSLLHSSGKTYLY, RMSNLAS, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY: (c) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15, and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 16; (d) the amino acid sequences of RSSKSLLHSSGKTYLY, RLSNLAS, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY; (e) the amino acid sequences of RSSKSLLHSSGKTYLY, RLSSNLAS, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY; (f) a variable light chain (VL) comprising the amino acid sequence of SEQ ID NO: 17, SEQ ID NO: 19, or SEQ ID NO: 21, and a variable heavy chain (Vh) comprising the amino acid sequence of SEQ ID NO: 23; (g) a variable light chain comprising the amino acid sequence of SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 33 or SEQ ID NO: 34; (h) the amino acid sequences of RSSKSLLHSSGKTYLY, RMSNLA, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY; (i) the amino acid sequences of RSSKSLLHSSGKTYLY, RLSNLA, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY; (j) the amino acid sequences of RSSKSLLHSSGKTYLY, RLSSLA, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY; (k) a variable light chain comprising the amino acid sequence of SEQ ID NO: 35, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 37; (l) a variable light chain comprising the amino acid sequence of SEQ ID NO: 39, SEQ ID NO: 41, or SEQ ID NO: 43, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 45 or SEQ ID NO: 47; (m) a variable light chain comprising the amino acid sequence of SEQ ID NO: 55, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 56 or SEQ ID NO: 57; or (n) the amino acid sequences of RSSKSLLHSSGKTYLYW, RMSNLA, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY; wherein the antibody or fragment thereof has a Kd less than 2.16 x 10"11 M. 2. An isolated antibody or fragment thereof that competitively inhibits the binding of an anti-CXCR5 antibody or fragment thereof to the extracellular domain of human CXCR5, wherein the anti-CXCR5 antibody or fragment thereof comprises a variable light chain comprising the amino acid sequence of SEQ ID NO: 32, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 33, wherein the antibody or fragment thereof has a KD less than 2.16 x 10'11 M. 3. An isolated antibody or fragment thereof that competitively inhibits the binding of an anti-CXCR5 antibody or fragment thereof to the extracellular domain of human CXCR5, wherein the anti-CXCR5 antibody or fragment thereof comprises the amino acid sequences of RSSKSLLHSSGKTYLY, RLSSLA, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY, wherein the antibody or fragment thereof has a KD less than 2.16 x 10"11 M. 4. The antibody or fragment thereof of any one of claims 1-3, wherein the antibody or fragment thereof further comprises one or more constant region domains. 5. The antibody or fragment thereof of any one of claims 1-3, wherein the antibody or fragment thereof further comprises a CHI, CH2, CH3, CL, or combinations thereof. 6. The antibody or fragment thereof of claim 4, wherein the one or more constant region domains are from an IgG antibody. 7. The antibody or fragment thereof of claim 6, wherein the IgG antibody is an IgG4 antibody. 8. The antibody or fragment thereof of any one of claims 1-3, wherein the antibody or fragment thereof is a single chain Fv antibody. 9. A pharmaceutical composition comprising a therapeutically effective amount of the antibody or fragment thereof of any one of claims 1-8 and a pharmaceutically acceptable carrier. 10. An isolated nucleic acid molecule encoding the antibody or fragment thereof of any one of claims 1-8. 11. A vector comprising the nucleic acid molecule of claim 10. 12. An isolated host cell comprising the vector of claim 11, excluding cells capable of generating a human being. 13. A method of treating a patient having a CXCR5 disease or condition, the method comprising administering to the patient a therapeutically effective amount of the antibody or fragment thereof of any one of claims 1-8 or of a pharmaceutical composition of claim 9. 14. The method of claim 13, wherein the CXCR5 disease or condition is selected from the group consisting of pancreatic cancer, colon cancer, bladder cancer, T cell leukemia, B cell leukemia, lupus, Sjoren's Syndrome, myasthenia gravis, multiple sclerosis, ulcerative colitis, rheumatoid arthritis, psoriatic arthritis, a chronic inflammatory disease, and a graft undergoing rejection. 15. A method of treating a patient having a disorder comprising CXCR5 positive cells, the method comprising administering to the patient a therapeutically effective amount of the antibody or fragment thereof of any one of claims 1-8 or of a pharmaceutical composition of claim 9. 16. The method of claim 15, wherein the disorder comprising CXCR5 positive cells is selected from the group consisting of pancreatic cancer, colon cancer, bladder cancer, T cell leukemia, B cell leukemia, lupus, Sjoren's Syndrome, myasthenia gravis, multiple sclerosis, ulcerative colitis, rheumatoid arthritis, psoriatic arthritis, a chronic inflammatory disease, and a graft undergoing rejection. 17. The method of claim 15, wherein at least one of the CXCR5 positive cells is a B cell. 18. The method of claim 15, wherein at least one of the CXCR5 positive cells is activated by CXCL13. 19. The method of claim 15, wherein at least one of the CXCR5 positive cells is a T cell. 20. The use of the antibody or fragment thereof of any one of claims 1-8 in the manufacture of a medicament for the treatment of a CXCR5 disease or condition in a patient. 21. The use according to claim 20, wherein the CXCR5 disease or condition is selected from the group consisting of pancreatic cancer, colon cancer, bladder cancer, T cell leukemia, B cell leukemia, lupus, Sjoren's Syndrome, myasthenia gravis, multiple sclerosis, ulcerative colitis, rheumatoid arthritis, psoriatic arthritis, a chronic inflammatory disease, and a graft undergoing rejection. 22. The use of the antibody or fragment thereof of any one of claims 1-8 in the manufacture of a medicament for the treatment of a disorder comprising CXCR5 positive cells. 23. The use according to claim 22, wherein the disorder comprising CXCR5 positive cells is selected from the group consisting of pancreatic cancer, colon cancer, bladder cancer, T cell leukemia, B cell leukemia, lupus, Sjoren's Syndrome, myasthenia gravis, multiple sclerosis, ulcerative colitis, rheumatoid arthritis, psoriatic arthritis, a chronic inflammatory disease, and a graft undergoing rejection. 24. The use according to claim 22, wherein at least one of the CXCR5 positive cells is a B cell. 25. The use according to claim 22, wherein at least one of the CXCR5 positive cells is activated by CXCL13. 26. The use according to claim 22, wherein at least one of the CXCR5 positive cells is a T cell. 27. A method of manufacturing the antibody or fragment thereof of any one of claims 1-8, the method comprising expressing the vector of claim 11 in a suitable host cell. 28. A kit comprising the antibody or fragment thereof of any one of claims 1-8 and instructions for use. 29. The antibody or fragment thereof according to any one of claims 1-8, the composition according to claim 9, the isolated nucleic acid according to claim 10, the vector according to claim 11, the isolated host cell according to claim 12, the method according to any one of claims 13 to 19 or 27, the use according to any one of claims 20 to 26, or the kit according to claim 28, substantially as hereinbefore described.

SANOFI

WATERMARK PATENT AND TRADE MARKS ATTORNEYS P32965AU02

Claims

THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS:

1. An isolated antibody or fragment thereof that competitively inhibits the binding of an anti-CXCR5 antibody or fragment thereof to the extracellular domain of human CXCR5, wherein the anti-CXCR5 antibody or fragment thereof comprises: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 11, and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 12; (b) the amino acid sequences of RSSKSLLHSSGKTYLY, RMSNLAS, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY: (c) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15, and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 16; (d) the amino acid sequences of RSSKSLLHSSGKTYLY, RLSNLAS, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY; (e) the amino acid sequences of RSSKSLLHSSGKTYLY, RLSSNLAS, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY; (f) a variable light chain (VL) comprising the amino acid sequence of SEQ ID NO: 17, SEQ ID NO: 19, or SEQ ID NO: 21, and a variable heavy chain (Vh) comprising the amino acid sequence of SEQ ID NO: 23; (g) a variable light chain comprising the amino acid sequence of SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 33 or SEQ ID NO: 34; (h) the amino acid sequences of RSSKSLLHSSGKTYLY, RMSNLA, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY; (i) the amino acid sequences of RSSKSLLHSSGKTYLY, RLSNLA, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY; (j) the amino acid sequences of RSSKSLLHSSGKTYLY, RLSSLA, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY; (k) a variable light chain comprising the amino acid sequence of SEQ ID NO: 35, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 37; (l) a variable light chain comprising the amino acid sequence of SEQ ID NO: 39, SEQ ID NO: 41, or SEQ ID NO: 43, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 45 or SEQ ID NO: 47; (m) a variable light chain comprising the amino acid sequence of SEQ ID NO: 55, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 56 or SEQ ID NO: 57; or (n) the amino acid sequences of RSSKSLLHSSGKTYLYW, RMSNLA, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY; wherein the antibody or fragment thereof has a Kd less than 2.16 x 10"11 M.
2. An isolated antibody or fragment thereof that competitively inhibits the binding of an anti-CXCR5 antibody or fragment thereof to the extracellular domain of human CXCR5, wherein the anti-CXCR5 antibody or fragment thereof comprises a variable light chain comprising the amino acid sequence of SEQ ID NO: 32, and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 33, wherein the antibody or fragment thereof has a KD less than 2.16 x 10'11 M.
3. An isolated antibody or fragment thereof that competitively inhibits the binding of an anti-CXCR5 antibody or fragment thereof to the extracellular domain of human CXCR5, wherein the anti-CXCR5 antibody or fragment thereof comprises the amino acid sequences of RSSKSLLHSSGKTYLY, RLSSLA, MQHLEYPYT, GFSLIDYGVN, VIWGDGTTY, and IVY, wherein the antibody or fragment thereof has a KD less than 2.16 x 10"11 M.
4. The antibody or fragment thereof of any one of claims 1-3, wherein the antibody or fragment thereof further comprises one or more constant region domains.
5. The antibody or fragment thereof of any one of claims 1-3, wherein the antibody or fragment thereof further comprises a CHI, CH2, CH3, CL, or combinations thereof.
6. The antibody or fragment thereof of claim 4, wherein the one or more constant region domains are from an IgG antibody.
7. The antibody or fragment thereof of claim 6, wherein the IgG antibody is an IgG4 antibody.
8. The antibody or fragment thereof of any one of claims 1-3, wherein the antibody or fragment thereof is a single chain Fv antibody.
9. A pharmaceutical composition comprising a therapeutically effective amount of the antibody or fragment thereof of any one of claims 1-8 and a pharmaceutically acceptable carrier.
10. An isolated nucleic acid molecule encoding the antibody or fragment thereof of any one of claims 1-8.
11. A vector comprising the nucleic acid molecule of claim 10.
12. An isolated host cell comprising the vector of claim 11, excluding cells capable of generating a human being.
13. A method of treating a patient having a CXCR5 disease or condition, the method comprising administering to the patient a therapeutically effective amount of the antibody or fragment thereof of any one of claims 1-8 or of a pharmaceutical composition of claim 9.
14. The method of claim 13, wherein the CXCR5 disease or condition is selected from the group consisting of pancreatic cancer, colon cancer, bladder cancer, T cell leukemia, B cell leukemia, lupus, Sjoren's Syndrome, myasthenia gravis, multiple sclerosis, ulcerative colitis, rheumatoid arthritis, psoriatic arthritis, a chronic inflammatory disease, and a graft undergoing rejection.
15. A method of treating a patient having a disorder comprising CXCR5 positive cells, the method comprising administering to the patient a therapeutically effective amount of the antibody or fragment thereof of any one of claims 1-8 or of a pharmaceutical composition of claim 9.
16. The method of claim 15, wherein the disorder comprising CXCR5 positive cells is selected from the group consisting of pancreatic cancer, colon cancer, bladder cancer, T cell leukemia, B cell leukemia, lupus, Sjoren's Syndrome, myasthenia gravis, multiple sclerosis, ulcerative colitis, rheumatoid arthritis, psoriatic arthritis, a chronic inflammatory disease, and a graft undergoing rejection.
17. The method of claim 15, wherein at least one of the CXCR5 positive cells is a B cell.
18. The method of claim 15, wherein at least one of the CXCR5 positive cells is activated by CXCL13.
19. The method of claim 15, wherein at least one of the CXCR5 positive cells is a T cell.
20. The use of the antibody or fragment thereof of any one of claims 1-8 in the manufacture of a medicament for the treatment of a CXCR5 disease or condition in a patient.
21. The use according to claim 20, wherein the CXCR5 disease or condition is selected from the group consisting of pancreatic cancer, colon cancer, bladder cancer, T cell leukemia, B cell leukemia, lupus, Sjoren's Syndrome, myasthenia gravis, multiple sclerosis, ulcerative colitis, rheumatoid arthritis, psoriatic arthritis, a chronic inflammatory disease, and a graft undergoing rejection.
22. The use of the antibody or fragment thereof of any one of claims 1-8 in the manufacture of a medicament for the treatment of a disorder comprising CXCR5 positive cells.
23. The use according to claim 22, wherein the disorder comprising CXCR5 positive cells is selected from the group consisting of pancreatic cancer, colon cancer, bladder cancer, T cell leukemia, B cell leukemia, lupus, Sjoren's Syndrome, myasthenia gravis, multiple sclerosis, ulcerative colitis, rheumatoid arthritis, psoriatic arthritis, a chronic inflammatory disease, and a graft undergoing rejection.
24. The use according to claim 22, wherein at least one of the CXCR5 positive cells is a B cell.
25. The use according to claim 22, wherein at least one of the CXCR5 positive cells is activated by CXCL13.
26. The use according to claim 22, wherein at least one of the CXCR5 positive cells is a T cell.
27. A method of manufacturing the antibody or fragment thereof of any one of claims 1-8, the method comprising expressing the vector of claim 11 in a suitable host cell.
28. A kit comprising the antibody or fragment thereof of any one of claims 1-8 and instructions for use.
29. The antibody or fragment thereof according to any one of claims 1-8, the composition according to claim 9, the isolated nucleic acid according to claim 10, the vector according to claim 11, the isolated host cell according to claim 12, the method according to any one of claims 13 to 19 or 27, the use according to any one of claims 20 to 26, or the kit according to claim 28, substantially as hereinbefore described. SANOFI WATERMARK PATENT AND TRADE MARKS ATTORNEYS P32965AU02