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 method of producing a CTP-modified coagulation Factor VII (FVII) polypeptide, comprising the step of attaching three to five chorionic gonadotrophin carboxy terminal peptides (CTPs) to the carboxy terminus of a coagulation FVII polypeptide, thereby producing a CTP-modified FVII polypeptide.
2. The method of claim 1 , wherein at least one CTP consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
3. The method of claim 1 , wherein at least one CTP is glycosylated.
4. The method of claim 1 , wherein at least one CTP is truncated.
5. The method of claim 1 , wherein at least one CTP is attached to said coagulation Factor VII polypeptide via a linker.
6. The method of claim 5 , wherein said linker is a peptide bond.
7. The method of claim 1 , wherein the sequence of said CTP-modified Factor VII (FVII) is selected from the group consisting of SEQ ID NO: 25 and SEQ ID NO: 46.
8. The method of claim 1 , wherein said CTP-modified coagulation Factor VII has a signal peptide.
9. The method of claim 8 , wherein said signal peptide is set forth in SEQ ID NO: 47.
10. The method of claim 1 , wherein said coagulation FVII polypeptide is an activated coagulation Factor VII (FVIIa) polypeptide.
11. The method of claim 10 , wherein the sequence of said CTP-modified activated coagulation Factor VII is set forth in SEQ ID NO. 46.
12. The method of claim 10 , wherein said activated coagulation FVII polypeptide is in the form of a disulfide-linked two chain heterodimer.
13. The method of claim 12 , 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.
14. The method of claim 1 , wherein said step of attaching comprises attaching three CTPs to the carboxy terminus of said FVII polypeptide.