Long-acting coagulation factors and methods of producing same
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.
1. A CTP-modified polypeptide consisting of a coagulation factor and three to five chorionic gonadotropin carboxy terminal peptides (CTPs) attached to the carboxy terminus of said coagulation factor, wherein optionally a signal peptide is attached to the amino terminus of said coagulation factor, wherein at least one CTP is optionally attached to the 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. The CTP-modified polypeptide of claim 1 , wherein the sequence of said CTP-modified Factor VII (FVII) or activated Factor VII (FVIIa) is set forth in SEQ ID NO: 46.
7. The CTP-modified polypeptide of claim 1 , wherein said polypeptide has a signal peptide.
8. The CTP-modified polypeptide of claim 7 , wherein said signal peptide is set forth in SEQ ID NO: 47.
9. The CTP-modified polypeptide of claim 7 , wherein the sequence of said CTP-modified FVII is set forth in SEQ ID No: 25.
10. A pharmaceutical composition comprising the CTP-modified polypeptide of claim 1 and a pharmaceutically acceptable carrier.
11. The CTP-modified polypeptide of claim 1 , wherein said activated FVII (FVIIa) polypeptide is in the form of a disulfide-linked two chain heterodimer, wherein said two chains comprise a light chain consisting of amino acids 1-152 and a heavy chain consisting of amino acids 153-490 of SEQ ID NO: 46.
12. A method of extending the biological half-life of a coagulation factor, comprising the step of attaching three to five 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 optionally a signal peptide is attached to the amino terminus of said coagulation factor, wherein at least one CTP is optionally attached to the coagulation factor via a linker, and wherein said coagulation factor is selected from Factor VII (FVII) and activated Factor VII (FVIIa).
13. The method of claim 12 , wherein the sequence of at least one CTP is selected from the group consisting of: SEQ ID NO: 1 and SEQ ID NO: 2.
14. The method of claim 12 , wherein at least one CTP is glycosylated.
15. The method of claim 12 , wherein at least one CTP is truncated.
16. The method of claim 12 , wherein said linker is a peptide bond.
17. The method of claim 12 , wherein the sequence of said CTP-modified Factor VII (FVII) or activated Factor VII (FVIIa) is set forth in SEQ ID NO: 46.
18. The method of claim 12 , wherein said polypeptide has a signal peptide.
19. The method of claim 18 , wherein said signal peptide is set forth in SEQ ID NO: 47.
20. The method of claim 18 , wherein the sequence of said CTP-modified FVII is set forth in SEQ ID No: 25.
21. The method of claim 12 , wherein said activated FVII (FVIIa) polypeptide is in the form of a disulfide-linked two chain heterodimer, wherein said two chains comprise a light chain consisting of amino acids 1-152 and a heavy chain consisting of amino acids 153-490 of SEQ ID NO: 46.
22. A method of improving the area under the curve (AUC) of a coagulation factor, comprising the step of attaching three to five chorionic gonadotropin carboxy terminal peptides (CTPs) to the carboxy terminus of said coagulation factor, thereby improving the AUC of said coagulation factor, wherein optionally a signal peptide is attached to the amino terminus of said coagulation factor, wherein at least one CTP is optionally attached to the coagulation factor via a linker, and wherein said coagulation factor is selected from Factor VII (FVII) and activated Factor VII (FVIIa).
23. The method of claim 22 , wherein the sequence of at least one CTP is selected from the group consisting of: SEQ ID NO: 1 and SEQ ID NO: 2.
24. The method of claim 22 , wherein at least one CTP is glycosylated.
25. The method of claim 22 , wherein at least one CTP is truncated.
26. The method of claim 22 , wherein said linker is a peptide bond.
27. The method of claim 22 , wherein the sequence of said CTP-modified Factor VII (FVII) or activated Factor VII (FVIIa) is set forth in SEQ ID NO: 46.
28. The method of claim 22 , wherein said polypeptide has a signal peptide.
29. The method of claim 28 , wherein said signal peptide is set forth in SEQ ID NO: 47.
30. The method of claim 29 , wherein the sequence of said CTP-modified FVII is set forth in SEQ ID No: 25.
31. The method of claim 22 , wherein said activated FVII (FVIIa) polypeptide is in the form of a disulfide-linked two chain heterodimer, wherein said two chains comprise a light chain consisting of amino acids 1-152 and a heavy chain consisting of amino acids 153-490 of SEQ ID NO: 46.
32. A method of reducing the dosing frequency of a coagulation factor, comprising the step of attaching three to five chorionic gonadotropin carboxy terminal peptides (CTPs) to the carboxy terminus of said coagulation factor, thereby reducing the dosing frequency of said coagulation factor, wherein optionally a signal peptide is attached to the amino terminus of said coagulation factor, wherein at least one CTP is optionally attached to the coagulation factor via a linker, and wherein said coagulation factor is selected from Factor VII (FVII) and activated Factor VII (FVIIa).
33. The method of claim 32 , wherein the sequence of at least one CTP is selected from the group consisting of: SEQ ID NO: 1 and SEQ ID NO: 2.
34. The method of claim 32 , wherein at least one CTP is glycosylated.
35. The method of claim 32 , wherein at least one CTP is truncated.
36. The method of claim 32 , wherein said linker is a peptide bond.
37. The method of claim 32 , wherein the sequence of said CTP-modified Factor VII (FVII) or activated Factor VII (FVIIa) is set forth in SEQ ID NO: 46.
38. The method of claim 32 , wherein said polypeptide has a signal peptide.
39. The method of claim 38 , wherein said signal peptide is set forth in SEQ ID NO: 47.
40. The method of claim 39 , wherein the sequence of said CTP-modified FVII is set forth in SEQ ID No: 25.
41. The method of claim 32 , wherein said activated FVII (FVIIa) polypeptide is in the form of a disulfide-linked two chain heterodimer, wherein said two chains comprise a light chain consisting of amino acids 1-152 and a heavy chain consisting of amino acids 153-490 of SEQ ID NO: 46.