Modified factor VII or factor VIIa polypeptides

Patent metadata

Field	Value
Publication number	US-9493757-B2
Application number	US-201414326786-A
Country	US
Kind code	B2
Filing date	Jul 9, 2014
Priority date	Apr 28, 2000
Publication date	Nov 15, 2016
Grant date	Nov 15, 2016

Abstract

The invention provides vitamin K-dependent polypeptides with enhanced membrane binding affinity. These polypeptides can be used to modulate clot formation in mammals. Methods of modulating clot formation in mammals are also described.

First claim

What is claimed is:

1. A Factor VII or Factor VIIa polypeptide comprising a modified GLA domain that enhances membrane binding affinity of said polypeptide relative to a corresponding native Factor VII or Factor VIIa polypeptide, said modified GLA domain comprising at least one amino acid substitution at position 33, 34, or 35, wherein amino acid positions of the Factor VII or Factor VIIa polypeptide are numbered according to SEQ ID NO:3.

2. The polypeptide of claim 1 , wherein a hydrophobic amino acid residue is substituted at position 33.

3. The polypeptide of claim 2 , wherein a phenylalanine, leucine or isoleucine residue is substituted at position 33.

4. The polypeptide of claim 1 , wherein a hydrophobic amino acid residue is substituted at position 34.

5. The polypeptide of claim 4 , wherein a phenylalanine, leucine or isoleucine residue is substituted at position 34.

6. The polypeptide of claim 1 , wherein an aspartic acid or glutamic acid residue is substituted at position 34.

7. The polypeptide of claim 6 , wherein a glutamic acid residue is substituted at position 34.

8. The polypeptide of claim 1 , wherein a hydrophobic amino acid residue is substituted at position 35.

9. The polypeptide of claim 8 , wherein a phenylalanine, leucine or isoleucine residue is substituted at position 35.

10. The polypeptide of claim 1 , further comprising an amino acid substitution at position 10 or 11.

11. The polypeptide of claim 10 , wherein a glutamine, asparagine, glutamic acid, or aspartic acid residue is substituted at position 10.

12. The polypeptide of claim 10 , wherein a glutamine residue is substituted at position 10.

13. The polypeptide of claim 1 , further comprising an amino acid substitution at position 32.

14. The polypeptide of claim 13 , wherein a glutamic acid residue is substituted at position 32.

15. The polypeptide of claim 1 , further comprising an amino acid substitution at position 28.

16. The polypeptide of claim 15 , wherein a phenylalanine or a glutamic acid residue is substituted at position 28.

17. The polypeptide of claim 16 , wherein a phenylalanine residue is substituted at position 28.

18. The polypeptide of claim 1 , further comprising an insertion at position 4.

19. The polypeptide of claim 18 , wherein a tyrosine or glycine residue is inserted at position 4.

20. The polypeptide of claim 19 , wherein a tyrosine residue is inserted at position 4.

21. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and an effective amount of a Factor VII or Factor VIIa polypeptide, wherein said Factor VII or Factor VIIa polypeptide comprises a modified GLA domain that enhances membrane binding affinity of said polypeptide relative to a corresponding native Factor VII or Factor VIIa polypeptide, said modified GLA domain comprising at least one amino acid substitution at position 33, 34, or 35, wherein amino acid positions of the Factor VII or Factor VIIa polypeptide are numbered according to SEQ ID NO:3.

22. The pharmaceutical composition of claim 21 , wherein the Factor VII or Factor VIIa polypeptide further comprises a glutamine residue substituted at position 10 and a glutamic acid residue substituted at position 32.

23. A method of increasing clot formation in a mammal comprising administering an amount of a Factor VII or Factor VIIa polypeptide effective to increase clot formation in said mammal, wherein said Factor VII or Factor VIIa polypeptide comprises a modified GLA domain that enhances membrane binding affinity of said polypeptide relative to a corresponding native Factor VII or Factor VIIa polypeptide, said modified GLA domain comprising at least one amino acid substitution at position 33, 34, or 35, wherein amino acid positions of the Factor VII or Factor VIIa polypeptide are numbered according to SEQ ID NO:3.

24. The method of claim 23 , wherein the mammal has a bleeding disorder.

Assignees

• Univ Minnesota

Inventors

• Nelsestuen Gary L

Classifications

• C12N9/64 Primary

  derived from animal tissue · CPC title

• C12N9/647

  Blood coagulation factors not provided for in a preceding group or according to more than one of the proceeding groups · CPC title

• C12Y304/21069

  Protein C activated (3.4.21.69) · CPC title

• C07K14/745 Primary

  Blood coagulation or fibrinolysis factors · CPC title

• C12Y304/21021

  Coagulation factor VIIa (3.4.21.21) · CPC title