Long-acting coagulation factors and methods of producing same

• US 9,458,444 B2
• Filed: 02/05/2013
• Issued: 10/04/2016
• Est. Priority Date: 02/03/2006
• Status: Active Grant

• Current Assignee: Opko Biologics Ltd. (Opko Health, Inc.)
• Original Assignee: Opko Biologics Ltd. (Opko Health, Inc.)
• Inventors: Fima, Udi Eyal, Hart, Gili
• Primary Examiner(s): Kemmerer, Elizabeth C
• Assistant Examiner(s): Deberry, Regina M
• Application Number:
  US13/759,860
• Publication Number:
  US 20130243747A1
• Time in Patent Office: 1,337 Days
• Field of Search: 530/384, 530/398, 530/402, 514/9.7, 514/13.7, 514/14.3
• US Class Current: 1/1
• 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: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: C07K 14/505 Erythropoietin [EPO]
• CPC Class Codes: C07K 14/59 Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g.hCG [human chorionic gonadotropin]; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
• CPC Class Codes: C07K 14/745 Blood coagulation or fibrinolysis factors
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C07K 2319/31 fusions, other than Fc, for prolonged plasma life, e.g. albumin
• CPC Class Codes: C07K 2319/91 containing a motif for glycosylation
• CPC Class Codes: C12N 9/64 derived from animal tissue
• CPC Class Codes: C12N 9/6437 Coagulation factor VIIa (3.4.21.21)
• CPC Class Codes: C12N 9/644 Coagulation factor IXa (3.4.21.22)
• CPC Class Codes: C12Y 304/21021 Coagulation factor VIIa (3.4.21.21)

Abstract

Polypeptides comprising at least one carboxy-terminal peptide (CTP) of chorionic gonadotrophin attached to the carboxy terminus but not to the amino terminus of a coagulation factor and polynucleotides encoding the same are disclosed. Pharmaceutical compositions comprising the polypeptides and polynucleotides of the invention and methods of using and producing same are also disclosed.

Claims

1. A CTP-modified polypeptide consisting of a coagulation factor and three chorionic gonadotropin carboxy terminal peptides (CTPs) attached to the carboxy terminus of said coagulation factor, wherein at least one CTP is optionally attached to said coagulation factor via a linker, and wherein said coagulation factor is selected from Factor VII (FVII) and activated Factor VII (FVIIa).

2. The CTP-modified polypeptide of claim 1, wherein the sequence of at least one CTP is selected from the group consisting of:

SEQ ID NO;

1 and SEQ ID NO;

2.

3. The CTP-modified polypeptide of claim 1, wherein at least one CTP is glycosylated.

4. The CTP-modified polypeptide of claim 1, wherein at least one CTP is truncated.

5. The CTP-modified polypeptide of claim 1, wherein said linker is a peptide bond.

6. A pharmaceutical composition comprising the CTP-modified polypeptide of claim 1 and a pharmaceutically acceptable carrier.

7. The CTP-modified polypeptide of claim 1, wherein the sequence of said CTP-modified coagulation factor is set forth in SEQ ID NO:

25.

8. The CTP-modified polypeptide of claim 1, wherein the sequence of said CTP-modified coagulation factor is set forth in SEQ ID NO:

46.

9. A method of extending the biological half-life of a coagulation factor, comprising the step of attaching three chorionic gonadotropin carboxy terminal peptides (CTPs) to the carboxy terminus of said coagulation factor, thereby extending the biological half-life of said coagulation factor, wherein at least one CTP is optionally attached to said coagulation factor via a linker, and wherein said coagulation factor is selected from Factor VII (FVII) and activated Factor VII (FVIIa).

10. The method of claim 9, wherein the sequence of at least one CTP is selected from the group consisting of:

SEQ ID NO;

1 and SEQ ID NO;

2.

11. The method of claim 9, wherein at least one CTP is glycosylated.

12. The method of claim 9, wherein at least one CTP is truncated.

13. The method of claim 9, wherein said linker is a peptide bond.

14. The method of claim 9, wherein the sequence of said CTP-modified coagulation factor is set forth in SEQ ID NO:

25.

15. The method of claim 9, wherein the sequence of said CTP-modified coagulation factor is set forth in SEQ ID NO:

46.

16. A method of improving the area under the curve (AUC) of a coagulation factor, comprising the step of attaching three chorionic gonadotropin carboxy terminal peptides (CTPs) to the carboxy terminus of said coagulation factor, thereby improving the AUC of said coagulation factor, wherein at least one CTP is optionally attached to said coagulation factor via a linker, and wherein said coagulation factor is selected from Factor VII (FVII) and activated Factor VII (FVIIa).

17. The method of claim 16, wherein the sequence of at least one CTP is selected from the group consisting of:

SEQ ID NO;

1 and SEQ ID NO;

2.

18. The method of claim 16, wherein at least one CTP is glycosylated.

19. The method of claim 16, wherein at least one CTP is truncated.

20. The method of claim 16, wherein said linker is a peptide bond.

21. The method of claim 16, wherein the sequence of said CTP-modified coagulation factor is set forth in SEQ ID NO:

25.

22. The method of claim 16, wherein the sequence of said CTP-modified coagulation factor is set forth in SEQ ID NO:

46.

23. A method of reducing the dosing frequency of a coagulation factor, comprising the step of attaching three chorionic gonadotropin carboxy terminal peptides (CTPs) to the carboxy terminus of said coagulation factor, thereby reducing the dosing frequency of said coagulation factor, wherein at least one CTP is optionally attached to said coagulation factor via a linker, and wherein said coagulation factor is selected from Factor VII (FVII) and activated Factor VII (FVIIa).

24. The method of claim 23, wherein the sequence of at least one CTP is selected from the group consisting of:

SEQ ID NO;

1 and SEQ ID NO;

2.

25. The method of claim 23, wherein at least one CTP is glycosylated.

26. The method of claim 23, wherein at least one CTP is truncated.

27. The method of claim 23, wherein said linker is a peptide bond.

28. The method of claim 23, wherein the sequence of said CTP-modified coagulation factor is set forth in SEQ ID NO:

25.

29. The method of claim 23, wherein the sequence of said CTP-modified coagulation factor is set forth in SEQ ID NO:

46.