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 Factor IX (FIX) polypeptide consisting of a FIX polypeptide and three chorionic gonadotropin carboxy terminal peptides (CTPs) attached to the carboxy terminus of said CTP-modified FIX polypeptide, wherein at least one CTP is optionally attached to said FIX polypeptide via a linker.
2. The CTP-modified FIX polypeptide of claim 1 , wherein at least one CTP is encoded by an amino acid sequence selected from the group consisting of: SEQ ID NO: 1 and SEQ ID NO: 2.
3. The CTP-modified FIX polypeptide of claim 1 , wherein at least one CTP is glycosylated.
4. The CTP-modified FIX polypeptide of claim 1 , wherein at least one CTP is truncated.
5. The CTP-modified FIX polypeptide of claim 1 , wherein said linker is a peptide bond.
6. A pharmaceutical composition comprising the CTP-modified FIX polypeptide of claim 1 .
7. A method of treating hemophilia B in a subject comprising administering the CTP-modified Factor IX (FIX) polypeptide of claim 1 to a subject.
8. A method of extending the biological half-life of a Factor IX (FIX) polypeptide, comprising the step of attaching three chorionic gonadotrophin carboxy terminal peptides (CTPs) to the carboxy terminus of said FIX polypeptide, wherein at least one CTP is optionally attached to said FIX polypeptide via a linker, thereby extending the biological half-life of said FIX polypeptide.
9. The method of claim 8 , wherein at least one CTP is encoded by an amino acid sequence selected from the group consisting of: SEQ ID NO: 1 and SEQ ID NO: 2.
10. The method of claim 8 , wherein at least one CTP is glycosylated.
11. The method of claim 8 , wherein at least one CTP is truncated.
12. The method of claim 8 , wherein said linker is a peptide bond.
13. A method of producing a CTP-modified Factor IX (FIX) polypeptide, comprising the step of attaching three chorionic gonadotrophin carboxy terminal peptides (CTPs) to the carboxy terminus of said FIX polypeptide, wherein at least one CTP is optionally attached to said FIX polypeptide via a linker, thereby producing a CTP-modified FIX polypeptide.
14. The method of claim 13 , wherein the sequence of said CTP-modified FIX polypeptide is the sequence set forth in SEQ ID NO: 31.
15. The method of claim 13 , wherein at least one CTP is encoded by an amino acid sequence selected from the group consisting of: SEQ ID NO: 1 and SEQ ID NO: 2.
16. The method of claim 13 , wherein at least one CTP is glycosylated.
17. The method of claim 13 , wherein at least one CTP is truncated.
18. The method of claim 13 , wherein said linker is a peptide bond.
19. A method of improving the area under the curve (AUC), reducing the dosing frequency, or reducing the clearance rate of a Factor IX (FIX) polypeptide, comprising the step of attaching three chorionic gonadotrophin carboxy terminal peptides (CTPs) to the carboxy terminus of said FIX polypeptide, wherein at least one CTP is optionally attached to said FIX polypeptide via a linker, thereby improving the area under the curve (AUC), reducing the dosing frequency, or reducing the clearance rate of said FIX polypeptide.
20. The method of claim 19 , wherein at least one CTP is encoded by an amino acid sequence selected from the group consisting of: SEQ ID NO: 1 and SEQ ID NO: 2.
21. The method of claim 19 , wherein at least one CTP is glycosylated.
22. The method of claim 19 , wherein at least one CTP is truncated.
23. The method of claim 19 , wherein said linker is a peptide bond.