Long-acting polypeptides and methods of producing same
A polypeptide and polynucleotides encoding same comprising one carboxy-terminal peptide (CTP) of chorionic gonadotrophin attached to an amino terminus of a cytokine and two carboxy-terminal peptides (CTP) of chorionic gonadotrophin attached to a carboxy terminus of a cytokine are disclosed. Pharmaceutical compositions comprising the polypeptide and polynucleotides of the invention and methods of using same are also disclosed.
1. A polypeptide consisting of a cytokine, one chorionic gonadotrophin carboxy terminal peptide (CTP) attached to the amino terminus of said cytokine, and two chorionic gonadotrophin carboxy terminal peptides attached to the carboxy terminus of said cytokine.
2. The polypeptide of claim 1 , wherein the sequence of at least one CTP consists of an amino acid sequence selected from the group consisting of: SEQ ID NO: 17 and SEQ ID NO: 18.
3. The polypeptide of claim 1 , wherein at least one CTP is glycosylated.
4. The polypeptide of claim 1 , wherein at least one CTP is truncated.
5. The polypeptide of claim 1 , wherein at least one CTP is attached to said cytokine via a linker.
6. The polypeptide of claim 5 , wherein said linker is a peptide bond.
7. A pharmaceutical composition comprising the polypeptide of claim 1 and a pharmaceutically acceptable carrier.
8. A polypeptide consisting of a cytokine, one chorionic gonadotrophin carboxy terminal peptide (CTP) attached to the amino terminus of said cytokine, two chorionic gonadotrophin carboxy terminal peptides attached to the carboxy terminus of said cytokine, and a signal peptide attached to the amino terminus of said one CTP.
9. The polypeptide of claim 8 , wherein the sequence of said signal peptide comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 19 and SEQ ID NO: 64.
10. The polypeptide of claim 8 , wherein the sequence of at least one CTP consists of an amino acid sequence selected from the group consisting of: SEQ ID NO: 17 and SEQ ID NO: 18.
11. The polypeptide of claim 8 , wherein at least one CTP is glycosylated.
12. The polypeptide of claim 8 , wherein at least one CTP is truncated.
13. The polypeptide of claim 8 , wherein at least one CTP is attached to said cytokine via a linker.
14. The polypeptide of claim 13 , wherein said linker is a peptide bond.
15. A method of extending the biological half life of a cytokine, consisting of the step of attaching one chorionic gonadotrophin carboxy terminal peptide (CTP) to the amino terminus of said cytokine and two chorionic gonadotrophin carboxy terminal peptides to the carboxy terminus of said cytokine, thereby extending the biological half life of a cytokine.
16. The method of claim 15 , wherein the sequence of at least one CTP consists of an amino acid sequence selected from the group consisting of: SEQ ID NO: 17 and SEQ ID NO: 18.
17. The method of claim 15 , wherein at least one CTP is glycosylated.
18. The method of claim 15 , wherein at least one CTP is truncated.
19. The method of claim 15 , wherein at least one CTP is attached to said cytokine via a linker.
20. A method of extending the biological half life of a cytokine, consisting of the step of attaching one chorionic gonadotrophin carboxy terminal peptide (CTP) to the amino terminus of said cytokine, two chorionic gonadotrophin carboxy terminal peptides to the carboxy terminus of said cytokine and a signal peptide to the amino terminus of said one CTP, thereby extending the biological half life of a cytokine.
21. The method of claim 20 , wherein the amino acid sequence of at least one CTP consists of an amino acid sequence selected from the group consisting of: SEQ ID NO: 17 and SEQ ID NO: 18.
22. The method of claim 20 , wherein the sequence of said signal peptide comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 19 and SEQ ID NO: 64.
23. The method of claim 20 , wherein at least one CTP is glycosylated.
24. The method of claim 20 , wherein at least one CTP is truncated.
25. The method of claim 20 , wherein at least one CTP is attached to said cytokine via a linker.