Tek antagonists

• US 20030162712A1
• Filed: 02/03/2003
• Published: 08/28/2003
• Est. Priority Date: 06/07/1999
• Status: Active Grant

• Current Assignee: Immunex Corporation (Amgen Inc.)
• Original Assignee: Immunex Corporation (Amgen Inc.)
• Inventors: Borges, Luis G., Fanslow, William C. III, Cerretti, Douglas P.
• Granted Patent: US 7,067,475 B2
• Time in Patent Office: Days
• US Class Current: 514/12
• 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: 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 Tek extracellular domain, wherein the fragment lacks all or part of the region containing fibronectin type III (FNIII) motifs and wherein the polypeptide retains the ability to bind at least one Tek ligand.

2. The polypeptide of claim 1, wherein the fragment lacks at least residues 473-745 of the Tek extracellular domain.

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

4. 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.

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

6. The polypeptide of claim 1 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 70% identical to (a);

(c) variants that are at least 80% identical to (a);

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

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

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

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

7. A nucleic acid encoding a polypeptide according to claim 1.

8. A nucleic acid selected from the group consisting of:

(a) nucleic acids that encode a polypeptide according to claim 6(a);

(b) nucleic acids that are at least 70%, 80%, 90%, 95%, 98%, or 99% identical to a nucleic acid of (a), and which encode a polypeptide capable of binding at least one Tek ligand; and

(c) nucleic acids that hybridize at moderate stringency to a nucleic acid of (a), and which encode a polypeptide capable of binding at least one Tek ligand.

9. A polypeptide encoded by the nucleic acid of claim 8.

10. The nucleic acid of claim 7, further encoding a signal peptide sequence.

11. The nucleic acid of claim 10 that encodes SEQ ID NO:

2.

12. A polypeptide produced by a process comprising expressing a nucleic acid according claim 8 in a recombinant host cell under conditions that permit expression of the polypeptide.

13. A polypeptide produced by a process comprising expressing a nucleic acid according to claim 10 in a recombinant host cell under conditions that permit expression of the polypeptide and collecting the polypeptide secreted from the host cell.

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

15. The soluble Tek multimer of claim 14 wherein the multimer is a dimer or trimer.

16. The soluble Tek multimer of claim 14 wherein the multimer comprises an Fc polypeptide or a leucine zipper.

17. The soluble Tek multimer of claim 14 comprising residues 23-472 of SEQ ID NO:

2.

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

2.

19. An antibody or antibody fragment that binds specifically to a polypeptide according to claim 1.

20. An antibody or antibody fragment that is capable of competitively inhibiting the binding of a Tek ligand to a polypeptide according to claim 1.

21. An antibody or antibody fragment that is capable of competitively inhibiting the binding of angiopoietin-2 to the soluble Tek multimer of claim 18.

22. The antibody or antibody fragment of claim 20, selected from the group consisting of monoclonal antibodies, humanized antibodies, transgenic antibodies, and human antibodies.

23. A method of inhibiting angiogenesis in a mammal in need of such treatment, comprising administering to the mammal an inhibition-effective amount of a polypeptide according to claim 1.

28. A method of inhibiting angiogenesis in a mammal in need of such treatment, comprising administering to the mammal an inhibition-effective amount of a soluble Tek multimer according to claim 16.

30. A method of inhibiting angiogenesis in a mammal in need of such treatment, comprising administering to the mammal an inhibition-effective amount of a soluble Tek multimer according to claim 18.

31. A method of inhibiting angiogenesis in a mammal in need of such treatment, comprising administering to the mammal an inhibition-effective amount of an antibody or antibody fragment according to claim 20.

42. A method of making a Tek antagonist comprising generating an antibody that binds specifically to a polypeptide according to claim 1.

43. A hybridoma or recombinant cell line that produces a monoclonal antibody according to claim 22.

24. A method of inhibiting angiogenesis in a mammal in need of such treatment, comprising administering to the mammal an inhibition-effective amount of a soluble Tek multimer.

25. The method of claim 24 wherein the multimer is a dimer or trimer.

26. The method of claim 24 wherein the soluble Tek multimer comprises an Fc polypeptide or a leucine zipper.

27. The method of claim 24 wherein the Tek is human Tek.

29. The method of claim 24 wherein the soluble Tek multimer comprises a polypeptide having a sequence selected from the group consisting of residues 23-977 of SEQ ID NO:

1, residues 23-745 of SEQ ID NO;

1, residues 23-704 of SEQ ID NO;

2, and residues 23-472 of SEQ ID NO;

2.

32. A method of inhibiting angiogenesis in a mammal in need of such treatment, comprising administering to the mammal an inhibition-effective amount of a compound selected from the group consisting of:

(a) 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 and wherein the polypeptide retains the ability to bind at least one Tek ligand;

(b) a soluble Tek multimer; and

(c) an antibody or antibody fragment that is capable of competitively inhibiting the binding of a Tek ligand to the polypeptide of (a);

wherein the polypeptide, soluble Tek multimer, antibody, or antibody fragment is administered in a composition comprising a pharmaceutically acceptable carrier.

33. The method of claim 32 wherein the mammal has a disease or condition mediated by angiogenesis.

34. The method of claim 33 wherein the disease or condition is characterized by ocular neovascularization.

35. The method of claim 33 wherein the disease or condition is a solid tumor.

36. The method of claim 32 wherein the method further comprises treating the mammal with a second chemotherapeutic agent.

37. The method of claim 32 wherein the method further comprises treating the mammal with radiation.

38. The method of claim 36 wherein the second chemotherapeutic agent is selected from the group consisting of alkylating agents, antimetabolites, vinca alkaloids and other plant-derived chemotherapeutics, nitrosoureas, antitumor antibiotics, antitumor enzymes, topoisomerase inhibitors, platinum analogs, adrenocortical suppressants, hormones, hormone agonists, hormone antagonists, antibodies, immunotherapeutics, blood cell factors, radiotherapeutics, and biological response modifiers.

39. The method of claim 36 wherein the second chemotherapeutic agent is selected from the group consisting of cisplatin, cyclophosphamide, mechloretamine, melphalan, bleomycin, carboplatin, fluorouracil, 5-fluorodeoxyuridine, methotrexate, taxol, asparaginase, vincristine, and vinblastine, lymphokines and cytokines such as interleukins, interferons (including alpha., beta, or delta), and TNF, chlorambucil, busulfan, carmustine, lomustine, semustine, streptozocin, dacarbazine, cytarabine, mercaptopurine, thioguanine, vindesine, etoposide, teniposide, dactinomycin, daunorubicin, doxorubicin, bleomycin, plicamycin, mitomycin, L-asparaginase, hydroxyurea, methylhydrazine, mitotane, tamoxifen, and fluoxymesterone.

40. The method of claim 36 wherein the second chemotherapeutic agent is selected from the group consisting of Flt3 ligand, CD40 ligand, interleukin-2, interleukin-12, 4-1BB ligand, anti-4-1BB antibodies, TNF antagonists and TNF receptor antagonists including TNFR/Fc, TWEAK antagonists and TWEAK-R antagonists including TWEAK-R/Fc, TRAIL, CD148 agonists, VEGF antagonists including anti-VEGF antibodies, and VEGF receptor antagonists.

41. A method of inhibiting the binding of a Tek ligand to Tek in a mammal in need of such treatment, comprising administering to the mammal an inhibition-effective amount of compound selected from the group consisting of:

(a) 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 and wherein the polypeptide retains the ability to bind at least one Tek ligand;

(b) a soluble Tek multimer; and

(c) an antibody or antibody fragment that is capable of competitively inhibiting the binding of a Tek ligand to the polypeptide of (a).