Tek antagonists comprising soluble tek extracellular binding domain

• US 6,413,932 B1
• Filed: 06/07/2000
• Issued: 07/02/2002
• Est. Priority Date: 06/07/1999
• Status: Expired due to Fees

• Current Assignee: Immunex Corporation (Amgen Inc.)
• Original Assignee: Immunex Corporation (Amgen Inc.)
• Inventors: Farnslow, William C. III, Borges, Luis G., Cerretti, Douglas P.
• Primary Examiner(s): Spector, Lorraine
• Assistant Examiner(s): Jiang, Dong
• Application Number:
  US09/590,656
• Time in Patent Office: 755 Days
• Field of Search: 530/350, 530/412, 530/402, 514/44, 514/2.21, 435/320.1, 435/440, 435/69.7, 435/183
• US Class Current: 514/7.6
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/71 for growth factors; for growth regulators
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence

Abstract

The present invention provides Tek antagonists and methods of inhibiting angiogenesis in a mammal by administering Tek antagonists. The methods are particularly useful in treating diseases or conditions mediated by angiogenesis, such as solid tumors and diseases or conditions characterized by ocular neovascularization.

Claims

1. A polypeptide comprising a fragment of the Tek extracellular domain shown as residues 19-745 of SEQ ID NO:

1 wherein the fragment lacks all or part of the region containing fibronectin type III (FNIII) motifs, wherein the fragment lacks at least the residues corresponding to residues 473-745 of SEQ ID NO;

1 and wherein the fragment retains the ability to bind at least one Tek ligand.

2. The polypeptide of claim 1, wherein the Tek ligand is angiopoietin-1, angiopoietin-2, or angiopoietin-4.

3. The polypeptide of claim 1, wherein said polypeptide has a higher binding affinity for a Tek ligand than does a polypeptide comprising full length Tek extracellular domain.

4. The polypeptide of claim 3, wherein said polypeptide has a higher binding affinity for angiopoietin-2 than does a polypeptide comprising full length Tek extracellular domain.

5. The polypeptide of claim 4 wherein the polypeptide has at least a five-fold higher binding affinity for angiopoietin-2.

6. The polypeptide of claim 4 wherein the polypeptide has at least a 21-fold higher binding affinity for angiopoietin-2.

7. The polypeptide of claim 6 further comprising an Fc polypeptide, a leucine zipper, or a peptide linker.

8. A composition comprising the polypeptide of claim 6 and a pharmaceutically acceptable carrier.

9. A soluble Tek multimer comprising at least one polypeptide according to claim 6.

10. The polypeptide of claim 1 further comprising an Fc polypeptide, a leucine zipper, or a peptide linker.

11. A composition comprising the polypeptide of claim 1 and a pharmaceutically acceptable carrier.

12. A soluble Tek multimer comprising at least one polypeptide according to claim 1.

13. The soluble Tek multimer of claim 12 wherein t multimer is a dimer or trimer.

14. A composition comprising the multimer of claim 12 and a pharmaceutically acceptable carrier.

15. The soluble Tek multimer of claim 6 wherein the multimer comprise an Fc polypeptide, a leucine zipper. peptide linker.

16. The soluble Tek multimeter of claim 15 comprising residues 23-704 of SEQ ID NO:

2.

17. The multimer of claim 15 wherein the multimer has at least a 21-fold higher binding affinity for angiopoietin-2 as compared to Tek745/Fc.

18. The soluble Tek multimeter of claim 12 comprising residues 23-472 of SEQ ID NO:

2.

19. The multimer of claim 18 wherein the multimer has at least a 21-fold higher binding affinity for angiopoietin-2 as compared to Tek745/Fc.

20. A composition comprising the multimer of claim 18 and a pharmaceutically acceptable carrier.

21. A polypeptide encoded by a nucleic acid selected from the group consisting of nucleic acids that are at least 90%, 95%, 98%, or 99% identical to a nucleic acid that encodes residues 23472 of SEQ ID NO:

2 or a fragment thereof, wherein the polypeptide is capable of binding at least one Tek ligand.

22. The polypeptide of claim 21 wherein the nucleic acid is at least 90% identical to a nucleic acid that encodes residues 23-472 of SEQ ID NO:

2 or a fragment thereof.

23. The polypeptide of claim 21 wherein the nucleic acid is at least 95% identical to a nucleic acid that encodes residues 23-472 of SEQ ID NO:

2 or a fragment thereof.

24. The polypeptide of claim 21 wherein the nucleic acid is at least 98% identical to a nucleic acid that encodes residues 23-472 of SEQ ID NO:

2 or a fragment thereof.

25. A polypeptide produced by a process comprising expressing a nucleic acid in a recombinant host cell under conditions that permit expression of the polypeptide, wherein the nucleic acid is selected from the group consisting of nucleic acids that are at least 90%, 95%, 98%, or 99% identical to a nucleic acid that encodes residues 23-472 of SEQ ID NO:

2 or a fragment thereof, and wherein the polypeptide is capable of binding at least one Tek ligand.

26. A polypeptide produced by a process comprising expressing a nucleic acid in a recombinant host cell under conditions that permit expression of the polypeptide, wherein the nucleic acid is selected from the group consisting of nucleic acids that are at least 90%, 95%, 98%, or 99% identical to a nucleic acid that encodes residues 23-472 of SEQ ID NO:

2 or a fragment thereof, optionally joined to a nucleic acid encoding a signal sequence, and wherein the polypeptide is capable of binding at least one Tek ligand.

27. A polypeptide comprising a fragment of Tek extracellular domain, wherein the fragment lacks all or part of the region containing fibronectin type III (FNIII) motifs, wherein the fragment retains the ability to bind at least one Tek ligand, and wherein the fragment of Tek extracellular domain is selected from the group consisting of:

(a) residues 23-472 of SEQ ID NO;

2 and fragments thereof that bind a Tek ligand;

(b) variants that are at least 90% identical to (a);

(c) variants that are at least 95% identical to (a);

(d) variants that are at least 98% identical to (a); and

(e) variants that are at least 99% identical to (a).

28. The polypeptide of claim 27 wherein the sequence of the Tek extracellular domain fragment is at least 95% identical to residues 23-472 of SEQ ID NO:

2 or to a fragment thereof that binds a Tek ligand.

29. The polypeptide of claim 27 wherein the sequence of the Tek extracellular domain fragment is at least 90% identical to residues 23-472 of SEQ ID NO:

2 or to a fragment thereof that binds a Tek ligand.

30. The polypeptide of claim 29 further comprising an Fc polypeptide, a leucine zipper, or a peptide linker.

31. A composition comprising the polypeptide of claim 29 and a pharmaceutically acceptable carrier.

32. A soluble Tek multimer comprising at least one polypeptide according to claim 29.

33. The polypeptide of claim 27 wherein the sequence of the Tek extracellular domain fragment is at least 98% identical to residues 23-472 of SEQ ID NO:

2 or to a fragment thereof that binds a Tek ligand.

34. The polypeptide of claim 33 further comprising an Fc polypeptide, a leucine zipper, or a peptide linker.

35. A composition comprising the polypeptide of claim 33 and a pharmaceutically acceptable carrier.

36. A soluble Tek multimer comprising at least one polypeptide according to claim 33.

37. The polypeptide of claim 27 wherein the sequence of the Tek extracellular domain fragment consists of residues 23-472 of SEQ ID NO:

2 or a fragment thereof that binds a Tek ligand.

38. A composition comprising the polypeptide of claim 37 and a pharmaceutically acceptable carrier.

39. The polypeptide of claim 27 comprising residues 23-704 of SEQ ID NO:

2.

40. The polypeptide of claim 27 further comprising an Fc polypeptide, a leucine zipper, or a peptide linker.

41. A composition comprising the polypeptide of claim 27 and a pharmaceutically acceptable carrier.

42. A soluble Tek multimer comprising at least one polypeptide according to claim 27.