Long-acting oxyntomodulin variants and methods of producing same
This invention is directed to a chorionic gonadotrophin carboxy terminal peptide (CTP) modified dual GLP-1/glucagon receptor agonist, and methods of producing and using the same. In one embodiment, the present invention provides a CTP-modified polypeptide comprising a dual GLP-1/glucagon receptor agonist and at least one chorionic gonadotrophin carboxy terminal peptide (CTP) attached to the amino terminus or carboxy terminus of the agonist.
1. A CTP-modified polypeptide comprising a dual GLP-1/glucagon receptor agonist and one to five chorionic gonadotropin carboxy terminal peptides (CTP) attached to the amino terminus or carboxy terminus of said agonist, wherein said CTP is from the beta-subunit of human chorionic gonadotropin and comprises amino acid positions 1-10 of SEQ ID NO:3.
2. The CTP-modified polypeptide of claim 1 , wherein said GLP-1/glucagon receptor agonist is oxyntomodulin.
3. The CTP-modified polypeptide of claim 1 , 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.
4. The CTP-modified polypeptide of claim 1 , wherein at least one CTP is glycosylated or truncated.
5. The CTP-modified polypeptide of claim 1 , wherein at least one CTP is attached to said agonist polypeptide via a linker.
6. The CTP-modified polypeptide of claim 5 , wherein said linker is a peptide bond.
7. The CTP-modified polypeptide of claim 1 , wherein two, three, four or five chorionic gonadotropin carboxy terminal peptides are attached to the carboxy terminus of said agonist.
8. The CTP-modified polypeptide of claim 1 , wherein three chorionic gonadotropin carboxy terminal peptides are attached to said agonist, one CTP on the amino terminus and two CTP on the carboxy terminus of said agonist.
9. The CTP-modified polypeptide of claim 1 , wherein two chorionic gonadotropin carboxy terminal peptides are attached to said agonist, one CTP on the carboxy terminus and one CTP on the amino terminus of said agonist.
10. The CTP-modified polypeptide of claim 1 , wherein two chorionic gonadotropin carboxy terminal peptides are attached to said agonist on the amino terminus of said agonist.
11. The CTP-modified polypeptide of claim 1 , wherein said CTP comprises the amino acid sequence set forth in SEQ ID NO: 3.
12. The CTP-modified polypeptide of claim 1 , wherein said polypeptide lacks a signal peptide.
13. The CTP-modified polypeptide of claim 1 , wherein the amino acid sequence of said CTP-modified polypeptide comprises SEQ ID NO: 9, 15,25, 27,29, amino acids 4-122 of SEQ ID NO: 21, or amino acids 1-119 of SEQ ID NO: 23.
14. The CTP-modified polypeptide of claim 13 , wherein said polypeptide is the mature form of said CTP-modified polypeptide and wherein said mature CTP-polypeptide lacks a signal peptide.
15. A pharmaceutical composition comprising the CTP-modified polypeptide of 8.
16. The pharmaceutical composition of claim 15 , wherein said GLP 1/glucagon receptor agonist is oxyntomodulin (OXM).
17. A method of producing a CTP-modified polypeptide comprising a dual GLP-1/glucagon receptor agonist and one to five chorionic gonadotropin carboxy terminal peptide (CTP) attached to said agonist, the method comprising the step of attaching one to five chorionic gonadotropin carboxy terminal peptides to the amino terminus or carboxy terminus of said agonist, wherein said CTP is from the beta-subunit of human chorionic gonadotropin and comprises amino acid positions 1-10 of SEQ ID NO:3.
18. The method of claim 17 , wherein said GLP-1/glucagon receptor agonist is oxyntomodulin.
19. The method of claim 17 , wherein two, three, four, or five chorionic gonadotropin carboxy terminal peptides are attached to the carboxy terminus of said agonist.
20. The method of claim 17 , wherein three chorionic gonadotropin carboxy terminal peptides are attached to said agonist, one CTP on the amino terminus and two CTP on the carboxy terminus of said agonist.
21. The method of claim 17 , wherein two chorionic gonadotropin carboxy terminal peptides are attached to said agonist, one CTP on the carboxy terminus and one CTP on the amino terminus of said agonist.
22. The method of claim 17 , wherein two chorionic gonadotropin carboxy terminal peptides are attached to said agonist on the amino terminus of said agonist.
23. The method of claim 17 , wherein the amino acid sequence of said one to five CTP is selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO:2.
24. The method of claim 17 , wherein at least one CTP is glycosylated or is truncated.
25. The method of claim 17 , wherein at least one CTP is attached to said agonist via a linker.
26. The method of claim 25 , wherein said linker is a peptide bond.
27. The method of claim 17 , wherein producing said CTP-modified polypeptide extends the biological half-life of said agonist.
28. The method of claim 17 , wherein producing said CTP-modified polypeptide improves the area under the curve (AUC) of said agonist.
29. The method of claim 17 , wherein producing said CTP-modified polypeptide reduces the dosing frequency of said agonist.
30. The method of claim 17 , wherein said CTP comprises the amino acid sequence set forth in SEQ ID NO: 3.
31. The method of claim 17 , wherein said CTP-modified polypeptide lacks a signal peptide.
32. The method of claim 17 , wherein the amino acid sequence of said CTP-modified polypeptide comprises SEQ ID NO: 9, 15, 25, 27, 29, amino acids 4-122 of SEQ ID NO: 21, or amino acids 1-119 of SEQ ID NO: 23.
33. The method of claim 32 , wherein said polypeptide is the mature form of said CTP-modified polypeptide and wherein said mature CTP-polypeptide lacks a signal peptide.