Novel inhibitors of vascular endothelial growth factor activity, their uses and processes for their production

• US 20030092604A1
• Filed: 03/20/2002
• Published: 05/15/2003
• Est. Priority Date: 05/07/1996
• Status: Active Grant

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Ferrara, Napoleone, Davis-Smyth, Terri Lynn, Presta, Leonard, Chen, Helen Hsifei
• Granted Patent: US 6,897,294 B2
• Time in Patent Office: Days
• US Class Current: 530/350
• 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: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 14/71 for growth factors; for growth regulators
• CPC Class Codes: C07K 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/31 fusions, other than Fc, for prolonged plasma life, e.g. albumin

Abstract

The present invention is directed to novel chimeric VEGF receptor proteins comprising amino acid sequences derived from the vascular endothelial growth factor (VEGF) receptors flt-1 and KDR, including the murine homologue to the human KDR receptor FLK-1, wherein said chimeric VEGF receptor proteins bind to VEGF and antagonize the endothelial cell proliferative and angiogenic activity thereof. The present invention is also directed to nucleic acids and expression vectors encoding these chimeric VEGF receptor proteins, host cells harboring such expression vectors, pharmaceutically acceptable compositions comprising such proteins, methods of preparing such proteins and to methods utilizing such proteins for the treatment of conditions associated with undesired vascularization.

Claims

1. A chimeric VEGF receptor protein being capable of binding to VEGF and thereby exerting an inhibitory effect thereon, wherein said chimeric VEGF receptor protein comprises immunoglobulin-like domains 1, 2 and 3 of the flt-1 and/or the KDR receptor or functional equivalents thereof.

2. The chimeric VEGF receptor protein according to claim 1 wherein immunoglobulin-like domains 1, 2 and 3 are all derived from the flt-1 receptor.

3. The chimeric VEGF receptor protein according to claim 1 wherein immunoglobulin-like domains 1, 2 and 3 are all derived from the KDR receptor.

4. The chimeric VEGF receptor protein according to claim 1 wherein immunoglobulin-like domains 1 and 3 are derived from the flt-1 receptor and immunoglobulin-like domain 2 is derived from the KDR receptor.

5. The chimeric VEGF receptor protein according to claim 1 wherein immunoglobulin-like domains 1 and 3 are derived from the KDR receptor and immunoglobulin-like domain 2 is derived from the flt-1 receptor.

6. The chimeric VEGF receptor protein according to claim 1 wherein immunoglobulin-like domains 1 and 2 are derived from the flt-1 receptor and immunoglobulin-like domain 3 is derived from the KDR receptor.

7. The chimeric VEGF receptor protein according to claim 1 wherein immunoglobulin-like domains 1 and 2 are derived from the KDR receptor and immunoglobulin-like domain 3 is derived from the flt-1 receptor.

8. The chimeric VEGF receptor protein according to claim 1 wherein immunoglobulin-like domain 1 is derived from the flt-1 receptor and immunoglobulin-like domains 2 and 3 are derived from the KDR receptor.

9. The chimeric VEGF receptor protein according to claim 1 wherein immunoglobulin-like domain 1 is derived from the KDR receptor and immunoglobulin-like domains 2 and 3 are derived from the flt-1 receptor.

10. The chimeric VEGF receptor protein according to claim 1 wherein said immunoglobulin-like domains 1, 2 and 3 are fused to the F_c portion of a human IgG immunoglobulin.

11. The chimeric VEGF receptor protein according to claim 1 which is soluble.

19. A nucleic acid encoding the chimeric VEGF receptor protein of claims 1 or 12.

20. A replicable expression vector capable in a transformant host cell of expressing the chimeric VEGF receptor protein of claims 1 or 12.

21. Host cells transformed with the replicable expression vector according to claim 20.

22. Host cells according to claim 21 which are CEN4 cells.

23. A composition of matter comprising a chimeric VEGF receptor protein according to claim 1 compounded with a pharmaceutically acceptable carrier.

24. A method of producing a chimeric VEGF receptor protein comprising the steps of introducing into a suitable expression system the expression vector of claim 20 and effecting the expression of said chimeric VEGF receptor protein.

25. A method of therapeutically treating a mammal for a condition associated with undesired vascularization, said method comprising administering to said mammal a therapeutically effective amount of a composition according to claim 23.

26. The method according to claim 25 wherein said condition associated with undesired vascularization includes the formation of a tumor.

27. The method according to claim 26 wherein said tumor is malignant.

28. A method of substantially inhibiting VEGF activity in a mammal comprising administering to said mammal a VEGF inhibiting amount of a chimeric VEGF receptor protein according to claims 1 or 12.

12. A chimeric VEGF receptor protein being capable of binding to VEGF and thereby exerting an inhibitory effect thereon, said chimeric VEGF receptor protein comprising the extracellular domain of the FLT4 receptor wherein immunoglobulin-like domain 2 of said FLT4 receptor is replaced by immunoglobulin-like domain 2 of the flt-1 or KDR receptor or a functional equivalent thereof and wherein one or more other immunoglobulin-like domains of said FLT4 receptor are or are not replaced by the corresponding immunoglobulin-like domain of the flt-1 or KDR receptor.

13. The chimeric VEGF receptor protein according to claim 12 having immunoglobulin-like domain 2 of the flt-1 receptor and immunoglobulin-like domains 1 and 3 to 7 of the FLT4 receptor.

14. The chimeric VEGF receptor protein according to claim 12 having immuinoglobulin-like domains 1 to 3 of the flt-1 receptor and immunoglobulin-like domains 4 to 7 of the FLT4 receptor.

15. The chimeric VEGF receptor protein according to claim 12 whose amino acid sequence is shown in FIG. 9.

16. The chimeric VEGF receptor protein according to claim 12 whose amino acid sequence is shown in FIG. 10.

17. The chimeric VEGF receptor protein according to claim 12 which is soluble.

18. The chimeric VEGF receptor protein according to claim 12 which is membrane-bound.