Methods for the treatment of thrombosis

• US 20080058256A1
• Filed: 02/06/2007
• Published: 03/06/2008
• Est. Priority Date: 10/01/1999
• Status: Abandoned Application

• Current Assignee: Amgen Inc.
• Original Assignee: Amgen Inc.
• Inventors: Mann, Michael, Li, Huimin, Boone, Thomas
• Application Number:
  US11/702,972
• Publication Number:
  US 20080058256A1
• Time in Patent Office: Days
• US Class Current: 514/12
• CPC Class Codes: A61K 38/1703 from vertebrates
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: C12N 9/6418 from snakes
• CPC Class Codes: C12N 9/6489 Metalloendopeptidases (3.4.24)

Abstract

A fibrinolytically active metalloproteinase polypeptide (called “novel acting thrombolytic”) which is useful for blood clot lysis in vivo and methods and materials for its production by recombinant expression are described.

Claims

1. A thrombolytically active variant of SEQ ID NO:

5, wherein the sequence of amino acid residues Gln-Gln-Arg at positions 1-3 of SEQ ID NO;

5 is substituted with an amino acid that facilitates kex-2 cleavage when the amino acid occurs on the C-terminal side of the kex-2 hydrolysis site, and the remainder of the amino acid sequence of SEQ ID NO;

5 is unchanged.

2. A fusion polypeptide comprising the thrombolytically active variant of claim 1 fused to a heterologous polypeptide sequence.

3. A composition comprising the thrombolytically active variant of claim 1, and one or more components selected from a pharmaceutically acceptable diluent, a pharmaceutically acceptable preservative, a pharmaceutically acceptable solubilizer, a pharmaceutically acceptable emulsifier, a pharmaceutically acceptable adjuvant, or a pharmaceutically acceptable carrier.

4. A method for treating thrombosis in a mammal comprising administering locally to a clot in a blood vessel of the mammal a thrombolytically effective amount of the composition of claim 3.

5. The method of claim 4, wherein the composition is administered as two or more doses.

6. The method of claim 4, wherein the mammal is a human.

7. A method for lysing a blood clot comprising contacting the blood clot with a thrombolytically effective amount of the composition of claim 3.

8. The method of claim 7, wherein the blood clot is contacted in vivo.

9. The method of claim 8, wherein the blood clot is contacted in vivo in a human.

10. The method of claim 7, wherein the blood clot is contacted in vitro.

11. A nucleic acid molecule comprising a coding sequence encoding the thrombolytically active variant of claim 1.

12. An expression vector comprising the nucleic acid molecule of claim 11, operatively linked to expression regulatory elements.

13. A host cell comprising the expression vector of claim 12.

14. The host cell of claim 13, wherein the cell is a yeast cell.

15. The host cell of claim 14, wherein the yeast is Pichia pastoris.

16. A method for producing a recombinant polypeptide comprising providing a population of host cells according to claim 13 and causing expression of the polypeptide.

17. A method for producing a recombinant polypeptide comprising providing a population of host cells according to claim 14 and causing expression of the polypeptide.

18. A method for producing a recombinant polypeptide comprising providing a population of host cells according to claim 15 and causing expression of the polypeptide.