Long-acting coagulation factors and methods of producing same
Polypeptides comprising at least one carboxy-terminal peptide (CTP) of chorionic gonadotropin 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 Factor IX coagulation factor and three chorionic gonadotropin carboxy terminal peptides (CTPs) attached to the carboxy terminus of said coagulation factor, wherein said polypeptide does not include an N-terminal propeptide.
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 attached via a peptide bond.
5. The CTP-modified polypeptide of claim 1 , wherein the sequence of said CTP-modified coagulation factor is set forth in SEQ ID NO: 48.
6. The CTP-modified polypeptide of claim 1 , wherein said CTP-modified Factor IX is activated and comprises a disulfide-linked two chain heterodimer structure.
7. A pharmaceutical composition comprising the CTP-modified polypeptide of claim 1 , and a pharmaceutically acceptable carrier.
8. A method of extending the biological half-life of a Factor IX (FIX) coagulation factor, comprising the step of attaching three chorionic gonadotropin carboxy terminal peptides (CTP) to the carboxy terminus of said FIX coagulation factor, thereby extending the biological half-life of said coagulation factor, wherein said coagulation factor does not include an N-terminal propeptide.
9. The method of claim 8 , wherein the amino acid sequence of at least one CTP is 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 attached via a peptide bond.
12. The method of claim 8 , wherein the sequence of said CTP-modified FIX coagulation factor is set forth in SEQ ID NO: 48.
13. A method of improving the area under the curve (AUC) of a Factor IX (FIX) coagulation factor, comprising the step of attaching three chorionic gonadotropin carboxy terminal peptides (CTP) to the carboxy terminus of said FIX coagulation factor, thereby improving the AUC of said coagulation factor, wherein said coagulation factor does not include an N-terminal propeptide.
14. The method of claim 13 , wherein the amino acid sequence of at least one CTP is selected from the group consisting of: SEQ ID NO:1 and SEQ ID NO: 2.
15. The method of claim 13 , wherein at least one CTP is glycosylated.
16. The method of claim 13 , wherein at least one CTP is attached via a peptide bond.
17. The method of claim 13 , wherein the sequence of said CTP-modified FIX coagulation factor is as set forth in SEQ ID NO: 48.
18. A method of reducing the dosing frequency of a Factor IX (FIX) coagulation factor, comprising the step of attaching three chorionic gonadotropin carboxy terminal peptides (CTP) to the carboxy terminus of said FIX coagulation factor, thereby reducing the dosing frequency of said coagulation factor, wherein said coagulation factor does not include an N-terminal propeptide.
19. The method of claim 18 , wherein the amino acid sequence of at least one CTP is selected from the group consisting of: SEQ ID NO:1 and SEQ ID NO: 2.
20. The method of claim 18 , wherein at least one CTP is glycosylated.
21. The method of claim 18 , wherein at least one CTP is attached to said coagulation factor via a peptide bond.
22. The method of claim 18 , wherein the sequence of said CTP-modified FIX coagulation factor is set forth in SEQ ID NO: 48.
23. A method of treating hemophilia B in a subject, the method comprising the step of administering a CTP-modified coagulation factor polypeptide of claim 1 to said subject, thereby treating hemophilia B in said subject.
24. The method of claim 23 , wherein said subject is a human adult.
25. The method of claim 23 , wherein said subject is a human child.
26. The method of claim 23 , wherein said administering is via a subcutaneous route or via an intravenous route.