TARGETED BINDING AGENTS DIRECTED TO uPAR AND USES THEREOF

• US 20080152587A1
• Filed: 04/09/2007
• Published: 06/26/2008
• Est. Priority Date: 04/10/2006
• Status: Active Grant

• Current Assignee: MedImmune Ltd. (Astrazeneca PLC)
• Original Assignee: Amgen Fremont Inc. (Amgen Inc.), Astrazeneca AB (Astrazeneca PLC)
• Inventors: Emery, Stephen Charles, Zhou, Qing, Elvin, Paul
• Granted Patent: US 8,026,344 B2
• Time in Patent Office: Days
• US Class Current: 424/1.53
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 27/00 Drugs for disorders of the senses
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

Targeted binding agents directed to the antigen uPAR and uses of such antibodies are described. In particular, fully human monoclonal antibodies directed to the antigen uPAR. Nucleotide sequences encoding, and amino acid sequences comprising, heavy and light chain immunoglobulin molecules, particularly sequences corresponding to contiguous heavy and light chain sequences spanning the framework regions and/or complementarity determining regions (CDR'"'"'s), specifically from FR1 through FR4 or CDR1 through CDR3. Hybridomas or other cell lines expressing such immunoglobulin molecules and monoclonal antibodies.

Claims

1. An isolated targeted binding agent that specifically binds to urokinase-type plasminogen activator receptor (uPAR) and inhibits binding of urokinase-type plasminogen activator (uPA) to uPAR.

2. The targeted binding agent of claim 1, wherein said targeted binding agent inhibits greater than 90% of plasminogen activation on U937 cells.

3. The targeted binding agent of claim 1, wherein said targeted binding agent inhibits plasminogen activation on U937 cells with an IC₅₀ of less than 0.08 μ

g/ml.

4. The targeted binding agent of claim 1, wherein said targeted binding agent inhibits greater than 80% of uPAR-mediated cell adhesion of U937 cells to vitronectin at a concentration of 2 μ

g/ml.

5. The targeted binding agent of claim 1, wherein said targeted binding agent inhibits greater than 90% of cell invasion through Matrigel™

at a concentration of 10 μ

g/ml.

6. The targeted binding agent of claim 1, wherein said targeted binding agent inhibits tumor growth and metastasis in a mammal.

7. The targeted binding agent of claim 1, wherein said targeted binding agent binds uPAR with a K_d of less than 500 picomolar (pM).

8. The targeted binding agent of claim 1, wherein said targeted binding agent binds uPAR with a K_d of less than 400 picomolar (pM).

9. The targeted binding agent of claim 1, wherein said targeted binding agent binds uPAR with a K_d of less than 300 picomolar (pM).

10. The targeted binding agent of claim 1, wherein said targeted binding agent binds uPAR with a K_d of less than 200 picomolar (pM).

11. The targeted binding agent of claim 1, wherein said targeted binding agent is a monoclonal antibody.

12. A targeted binding agent according to claim 1, wherein said targeted binding agent is a monoclonal antibody selected from the group consisting of:

1.4.2, 1.41.1, 1.61.1, 1.99.1, 1.100.1, 1.113.2, 2.6.1, 2.19.2, 3.8.3, 3.176.2, 4.12.1, 4.13.1, 4.18.1, 4.19.1 and 4.50.1.

13. The targeted binding agent of claim 12, wherein said targeted binding agent is monoclonal antibody 2.19.2 (ATCC Accession Number PTA-7474).

14. The targeted binding agent of claim 12, wherein said targeted binding agent is monoclonal antibody 2.6.1 (ATCC Accession Number PTA-7475).

15. The targeted binding agent of claim 12, wherein said targeted binding agent is monoclonal antibody 4.18.1 (ATCC Accession Number PTA-7476).

17. A targeted binding agent, wherein said targeted binding agent is derivable from the monoclonal antibody of claim 12.

18. The targeted binding agent of claim 11, wherein said targeted binding agent is a fully human monoclonal antibody.

19. The targeted binding agent of claim 18, wherein said targeted binding agent is a binding fragment of a fully human monoclonal antibody.

20. The targeted binding agent of claim 19, wherein said binding fragment is selected from the group consisting of Fab, Fab′

, F(ab′

)₂, Fv and Dab.

21. The targeted binding agent of claim 1, wherein said targeted binding agent comprises a polypeptide having the sequence of SEQ ID NO.:

26.

22. The targeted binding agent of claim 21, wherein SEQ ID NO.:

26 has any one of the combinations of germline and non-germline residues indicated by each row of Table 15.

23. The targeted binding agent of claim 1, wherein said targeted binding agent comprises a polypeptide having the sequence of SEQ ID NO.:

28.

24. The targeted binding agent of claim 23, wherein SEQ ID NO.:

28 has any one of the combinations of germline and non-germline residues indicated by each row of Table 14.

25. The targeted binding agent of claim 1, wherein said targeted binding agent comprises a polypeptide having the sequence of SEQ ID NO.:

30.

26. The targeted binding agent of claim 25, wherein SEQ ID NO.:

30 has any one of the combinations of germline and non-germline residues indicated by each row of Table 13.

27. The targeted binding agent of claim 1, wherein said targeted binding agent comprises a polypeptide having the sequence of SEQ ID NO.:

32.

28. The targeted binding agent of claim 27, wherein SEQ ID NO.:

32 has any one of the combinations of germline and non-germline residues indicated by each row of Table 12.

29. The targeted binding agent of claim 1, wherein said targeted binding agent comprises a polypeptide having the sequence of SEQ ID NO.:

50.

30. The targeted binding agent of claim 29, wherein SEQ ID NO.:

50 has any one of the combinations of germline and non-germline residues indicated by each row of Table 17.

31. The targeted binding agent of claim 1, wherein said targeted binding agent comprises a polypeptide having the sequence of SEQ ID NO.:

52.

32. The targeted binding agent of claim 31, wherein SEQ ID NO.:

52 has any one of the combinations of germline and non-germline residues indicated by each row of Table 16.

39. A nucleic acid molecule encoding the targeted binding agent of claim 1.

40. A vector comprising the nucleic acid molecule of claim 39.

41. A host cell comprising the vector of claim 40.

42. A method of treating a malignant tumor in an animal, comprising:

selecting an animal in need of treatment for a malignant tumor; and

administering to said animal a therapeutically effective dose of the targeted binding agent of claim 1.

43. The method of claim 42, wherein said animal is human.

44. The method of claim 42, wherein said targeted binding agent is selected from the group consisting of 2.19.2 (ATCC Accession Number PTA-7474), 2.6.1 (ATCC Accession Number PTA-7475) and 4.18.1 (ATCC Accession Number PTA-7476).

45. The method of claim 41, wherein said malignant tumor is selected from the group consisting of:

melanoma, small cell lung cancer, non-small cell lung cancer, glioma, hepatocellular (liver) carcinoma, thyroid tumor, gastric (stomach) cancer, prostate cancer, breast cancer, ovarian cancer, bladder cancer, lung cancer, glioblastoma, endometrial cancer, kidney cancer, colon cancer, pancreatic cancer, esophageal carcinoma, head and neck cancers, mesothelioma, sarcomas, biliary (cholangiocarcinoma), small bowel adenocarcinoma, pediatric malignancies and epidermoid carcinoma.

46. A method of treating uPAR induced disease-related cell adhesion and/or invasion, comprising:

selecting an animal in need of treatment for disease-related cell adhesion and/or invasion; and

administering to said animal a therapeutically effective dose of the targeted binding agent of claim 1.

47. The method of claim 46, wherein said animal is human.

48. The method of claim 46, wherein said targeted binding agent is a fully human monoclonal antibody.

49. The method of claim 46, wherein said disease-related cell adhesion and/or invasion is tumor metastasis.

50. The method of claim 46, wherein said targeted binding agent is selected from the group consisting of 2.19.2 (ATCC Accession Number PTA-7474), 2.6.1 (ATCC Accession Number PTA-7475) and 4.18.1 (ATCC Accession Number PTA-7476).

51. A method of treating a non-neoplastic disease, comprising:

selecting an animal in need of treatment for a non-neoplastic disease; and

administering to said animal a therapeutically effective dose of the targeted binding agent of claim 1.

52. The method of claim 51, wherein said animal is human.

53. The method of claim 51, wherein said targeted binding agent is a fully human monoclonal antibody.

54. The method of claim 51, wherein said targeted binding agent is selected from the group consisting of 2.19.2 (ATCC Accession Number PTA-7474), 2.6.1 (ATCC Accession Number PTA-7475) and 4.18.1 (ATCC Accession Number PTA-7476).

55. The method of claim 51, wherein the non-neoplastic disease is selected from the group consisting of arthritis, atherosclerosis and allergic conjunctivitis.

56. A conjugate comprising the targeted binding agent of claim 1 or a binding fragment thereof, and a therapeutic agent.

57. The conjugate of claim 56, wherein the therapeutic agent is a toxin.

58. The conjugate of claim 56, wherein the therapeutic agent is a radioisotope.

59. The conjugate of claim 56, wherein the therapeutic agent is a pharmaceutical composition.

60. The targeted binding agent of claim 21, derived from a germline sequence with VH3-53, D6-19 and JH6B domains, wherein one or more residues has been mutated to yield the corresponding germline residue at that position.

61. The targeted binding agent of claim 23, derived from a germline sequence with A30 and JK4 domains, wherein one or more residues has been mutated to yield the corresponding germline residue at that position.

62. The targeted binding agent of claim 25, derived from a germline sequence with VH3-33, D2-21 and JH4B domains, wherein one or more residues has been mutated to yield the corresponding germline residue at that position.

63. The targeted binding agent of claim 27, derived from a germline sequence with B3 and JK1 domains, wherein one or more residues has been mutated to yield the corresponding germline residue at that position.

64. The targeted binding agent of claim 29, derived from a germline sequence with VH3-53, D6-19 and JH4B domains, wherein one or more residues has been mutated to yield the corresponding germline residue at that position.

65. The targeted binding agent of claim 31, derived from a germline sequence with L2 and JK5 domains, wherein one or more residues has been mutated to yield the corresponding germline residue at that position.

16. A targeted binding agent, wherein said targeted binding agent is a monoclonal antibody produced by the hybridoma selected from ATCC Accession Number PTA-7474, ATCC Accession Number PTA-7475 and ATCC Accession Number PTA-7476.

33. A targeted binding agent which cross-competes with any one of fully human monoclonal antibodies 2.19.2, 2.6.1 and 4.18.1 for binding to uPAR.

34. A targeted binding agent that binds to the same epitope on uPAR as any one of fully human monoclonal antibodies 2.19.2, 2.6.1 and 4.18.1.

35. A targeted binding agent comprising an amino acid sequence comprising

a) any one of a CDR1, CDR2 or CDR3 sequence as shown in Table 18 or Table 19;

b) a CDR1, CDR2 and CDR3 sequence as shown in Table 18;

c) a CDR1, CDR2 and CDR3 sequence as shown in Table 19;

or

d) a CDR1, CDR2 and CDR3 sequence as shown in Table 18 and a

CDR1, CDR2 and CDR3 sequence as shown in Table 19.

36. A targeted binding agent according to claim 35, wherein said targeted binding agent is an antibody.

37. A targeted binding agent according to claim 36, wherein said targeted binding agent is a fully human monoclonal antibody.

38. A targeted binding agent according to claim 37, wherein said targeted binding agent is a binding fragment of a fully human monoclonal antibody.