VON WILLEBRAND FACTOR (VWF)-CONTAINING PREPARATIONS, AND METHODS, KITS, AND USES RELATED THERETO

• US 20120289468A1
• Filed: 11/12/2010
• Published: 11/15/2012
• Est. Priority Date: 11/13/2009
• Status: Active Grant

• Current Assignee: Grifols Therapeutics LLC (Grifols SA)
• Original Assignee: Grifols Therapeutics Incorporated (Grifols SA)
• Inventors: Barnett, Thomas
• Granted Patent: US 8,759,293 B2
• Time in Patent Office: Days
• US Class Current: 514/14.1
• 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: A61P 7/04 Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: C07K 14/755 Factors VIII , e.g. factor VIII C (AHF), factor VIII Ag (VWF)

Abstract

The present invention relates to methods, compositions and kits for preparing FVIII and employing same. Also provided are vWF polypeptides and nucleic acid molecules encoding same.

Claims

1. A polypeptide comprising a first amino acid sequence present in a vWF polypeptide and a second amino acid sequence heterologous to the first, wherein the polypeptide is capable of binding a FVIII, with the proviso that the polypeptide lacks vWF domain A1, A2, A3, D4, B1, B2, B3, C1, C2, CK, or a combination thereof.

2. The polypeptide of claim 1, wherein the first amino acid sequence is as set forth in SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

33, SEQ ID NO;

34, or SEQ ID NO;

35.

3. The polypeptide of claim 1, wherein the polypeptide comprises the sequence set forth in SEQ ID NO:

17, SEQ ID NO;

18, SEQ ID NO;

19, SEQ ID NO;

20, SEQ ID NO;

21, SEQ ID NO;

22, SEQ ID NO;

36, SEQ ID NO;

38 or SEQ ID NO;

39

4. The polypeptide of claim 1, wherein the polypeptide is capable of forming a dimer.

5. The polypeptide of claim 1, wherein the second amino acid sequence corresponds to an immunoglobulin Fc.

6. The polypeptide of claim 1, wherein the second amino acid sequence has the sequence set forth in SEQ ID NO:

16.

7. A composition comprising the polypeptide of claim 1.

8. A protein complex comprising the polypeptide of claim 1 and the FVIII.

9. A composition comprising the protein complex of claim 8.

10. A nucleotide sequence encoding the polypeptide of claim 1.

11. The nucleotide sequence of claim 10, wherein the nucleotide sequence is set forth in SEQ ID NO:

23, SEQ ID NO;

24, SEQ ID NO;

25, SEQ ID NO;

26, SEQ ID NO;

27, SEQ ID NO;

28, SEQ ID NO;

37, SEQ ID NO;

42, or SEQ ID NO;

43.

12. An expression vector comprising the nucleotide sequence of claim 10.

13. A cell expressing the polypeptide of claim 1.

14. A cell expressing the protein complex of claim 8.

15. A method for preparing the protein complex of claim 8, the method comprising contacting the polypeptide with the FVIII.

16. The method of claim 15, wherein the contacting comprises recombinantly expressing the polypeptide in a cell that expresses the FVIII.

17. A method for preparing a FVIII, the method comprising:

contacting the FVIII with the polypeptide of claim 1 to form a protein complex comprising the polypeptide and the FVIII.

18. The method of claim 17 further comprising selectively adhering the complex to a resin comprising a binding partner having affinity for a domain defined by the second amino acid sequence.

19. The method of claim 18, wherein the second amino acid sequence corresponds to an immunoglobulin Fc.

20. The method of claim 19, wherein the binding partner is a protein A or a protein G.

21. A method for enhancing a plasma pharmacokinetic property of a FVIII, the method comprising administering to a subject a composition comprising the protein complex of claim 8.

22. The method of claim 21, wherein the property is extended plasma half-life.

23. A composition comprising the protein complex of claim 8, and a pharmaceutically acceptable carrier.

24. A method for treating a blood condition, the method comprising administering the composition of claim 23 to a subject.

25. The method of claim 24, wherein the condition is hemophilia.