Peptides binding to vascular endothelial growth factor

• US 20090035317A1
• Filed: 01/14/2008
• Published: 02/05/2009
• Est. Priority Date: 01/12/2007
• Status: Abandoned Application

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Kenrick, Sophia, Daugherty, Patrick S.
• Application Number:
  US12/008,828
• Publication Number:
  US 20090035317A1
• Time in Patent Office: Days
• US Class Current: 514/1.100
• 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 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/001 by chemical synthesis
• CPC Class Codes: C07K 14/52 Cytokines; Lymphokines; Interferons
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: G01N 2333/49 Platelet-derived growth factor [PDGF]
• CPC Class Codes: G01N 2500/04 Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)
• CPC Class Codes: G01N 33/74 involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors

Abstract

The invention relates to peptides that bind to vascular endothelial growth factor (VEGF). The invention further relates to methods of screening for VEGF-binding peptides, methods of using VEGF-binding peptides to detect the presence of VEGF in a biological sample, and methods of using VEGF-binding peptides to modulate VEGF activity and angiogenesis, and as components in a therapeutic or prophylactic composition.

Claims

3. A peptide comprising a W-E/D-W-E/D motif that binds to vascular endothelial growth factor (VEGF).

1. The peptide according to claim 3, comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:

1-30 and 32-51.

2. The peptide of claim 1 consisting of an amino acid sequence selected from the group consisting of SEQ ID NOS:

1-30 and 32-51.

4. The peptide of claim 3, wherein said peptide binds to VEGF with a dissociation constant (K_D) of less than 10⁻

6 M.

5. The peptide of claim 4, wherein said peptide binds to VEGF with a dissociation constant (K_D) of less than 10⁻

8 M.

6. The peptide of claim 5, wherein said peptide binds to VEGF with a dissociation constant (K_D) of less than 10⁻

10 M.

7. The peptide of claim 6, wherein said peptide binds to VEGF with a dissociation constant (K_D) of less than 10⁻

12 M.

8. The peptide of claim 3 comprising a sequence selected from the group consisting of SEQ ID NOS:

28-30.

9. The peptide of claim 3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:

1-30 and 32-51.

10. The peptide of claim 3 comprising the amino acid sequence of SEQ ID NO:

18.

11. The peptide of claim 3 comprising the amino acid sequence of SEQ ID NO:

24.

12. The peptide of claim 3, wherein the peptide is a bidentate peptide comprising a first VEGF-binding peptide sequence and a second VEGF-binding peptide sequence.

13. The peptide of claim 12, wherein the first peptide sequence comprises a sequence selected from the group consisting of SEQ ID NOS:

1-31, and the second peptide sequence comprises a sequence selected from the group consisting of SEQ ID NOS;

42-51.

14. The peptide of claim 13, wherein the bidentate peptide comprises a sequence selected from the group consisting of SEQ ID NOS:

32-41.

15. The peptide of claim 12, wherein the first and second VEGF-binding peptides are joined by a linker of two or more residues.

16. The peptide of claim 15, wherein the linker comprises the sequence of SEQ ID NO:

52.

17. A composition comprising at least one peptide according to claim 3.

18. The composition of claim 17, further comprising a pharmaceutically acceptable excipient.

19. The composition of claim 17, further comprising a vascular endothelial growth factor (VEGF)/angiogenesis inhibitor.

20. The composition of claim 19, wherein the VEGF/angiogenesis inhibitor is selected from the group consisting of an anti-VEGF/VEGFR antibody, a VEGF receptor tyrosine kinase inhibitor, a VEGF/VEGFR binding aptamer, an antisense inhibitor of VEGF/VEGFR expression, an anti-VEGF/VEGFR ribozyme, an anti-VEGF/VEGFR siRNA, and an endogenous VEGF/angiogenesis inhibitor.

21. The composition of claim 17, further comprising an anti-cancer agent.

22. The composition of claim 17, further comprising an anti-sepsis agent.

26. A method of treating a disorder selected from the group consisting of an angiogenesis disorder, cancer, and sepsis, the method comprising administering to a subject a therapeutically effective amount of the composition of claim 17.

27. The method of claim 26, wherein the peptide is administered prophylactically.

28. The method of claim 26, wherein administration of the peptide delays the onset of one or more symptoms of the disorder.

29. The method of claim 26, wherein administration of the peptide inhibits angiogenesis.

30. The method of claim 26, wherein administration of the peptide inhibits tumor growth.

31. The method of claim 26, wherein administration of the peptide inhibits infection.

32. A method for modulating angiogenesis in a subject comprising administering a therapeutically effective amount of the composition of claim 17.

33. The method of claim 32, wherein the composition inhibits angiogenesis in the subject.

34. The method of claim 32, wherein the composition is administered intermittently.

35. The method of claim 33, wherein the method further comprises administering to the subject a therapeutically effective amount of a vascular endothelial growth factor (VEGF)/angiogenesis inhibitor.

36. The method of claim 35, wherein the VEGF/angiogenesis inhibitor is selected from the group consisting of an anti-VEGF/VEGFR antibody, a VEGF receptor tyrosine kinase inhibitor, a VEGF/VEGFR binding aptamer, an antisense inhibitor of VEGF/VEGFR expression, an anti-VEGF/VEGFR ribozyme, an anti-VEGF/VEGFR siRNA, and an endogenous VEGF/angiogenesis inhibitor.

37. A method for inhibiting VEGF activity in a subject comprising administering a therapeutically effective amount of the composition of claim 17.

38. The method of claim 37, wherein the subject has cancer.

39. The method of claim 38, wherein the method further comprises administering to the subject a therapeutically effective amount of an anti-cancer agent.

40. The method claim 37, wherein the subject has sepsis.

41. The method of claim 40, wherein the method further comprises administering to the subject a therapeutically effective amount of an anti-sepsis agent.

42. The method of claim 41, wherein the composition is administered intermittently.

43. A method for detecting the presence of VEGF in a biological sample, the method comprising:

a) exposing the biological sample suspected of containing VEGF to the peptide of claim 3; and

b) detecting the presence or absence of the peptide bound to the VEGF, if any, in the sample.

44. The method of claim 43, wherein the peptide comprises a detectable label.

45. The method of claim 43, wherein the biological sample is obtained from a subject who has an angiogenesis disorder, cancer, or sepsis.

46. The method of claim 45, further comprising comparing the amount of VEGF in the biological sample from the subject who has an angiogenesis disorder, cancer or sepsis to the amount of VEGF in a corresponding biological sample from a normal subject.

47. The method of claim 43, further comprising collecting a plurality of biological samples from a subject at different time points and comparing the amount of VEGF in each biological sample to determine if the amount of VEGF is increasing or decreasing in the subject over time.

48. A complex comprising the peptide of claim 3 and VEGF.

49. The complex of claim 48 comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:

1-30 and 32-51.

23. A method of identifying a peptide comprising a W-E/D-W-E/D motif that binds to VEGF, the method comprising:

a) providing a library of peptides wherein each peptide in the library comprises the W-E/D-W-E/D motif; and

b) screening the library for peptides that bind to VEGF.

24. The method of claim 23, wherein the library is screened for peptides that bind to VEGF by sequential magnetic-activated cell sorting (MACS).

25. The method of claim 24, further comprising screening the library for peptides that bind to VEGF by fluorescence-activated cell sorting (FACS).