Long-acting polypeptides and methods of producing and administering same
A polypeptide and polynucleotides comprising at least two carboxy-terminal peptides (CTP) of chorionic gonadotrophin attached to a non-human peptide-of-interest are disclosed. Pharmaceutical compositions comprising the non-human polypeptides and polynucleotides of the invention and methods of using both human and non-human polypeptides and polynucleotides are also disclosed.
1. A method of treating an erythropoietin (EPO)-associated condition, wherein said EPO-associated condition is anemia, said method comprising administering to a subject a therapeutically effective amount of a polypeptide consisting of a chorionic gonadotropin carboxy terminal peptide (CTP) modified erythropoietin (EPO), said CTP-modified EPO polypeptide consisting of one chorionic gonadotropin carboxy terminal peptide (CTP) attached to the amino terminus of said EPO, and two chorionic gonadotropin CTPs attached to the carboxy terminus of said EPO, or said CTP-modified EPO polypeptide consisting of one chorionic gonadotropin carboxy terminal peptide (CTP) attached to the amino terminus of said EPO, and two chorionic gonadotropin CTPs attached to the carboxy terminus of said EPO, and a signal peptide attached to the amino terminal CTP.
2. The method of claim 1 , wherein the sequence of at least one chorionic gonadotrophin carboxy terminal peptide consists of an amino acid sequence selected from the group consisting of SEQ ID NO:17 and SEQ ID NO:18.
3. The method of claim 1 , wherein at least one chorionic gonadotrophin carboxy terminal peptide is truncated.
4. The method of claim 1 , wherein said EPO is glycosylated.
5. The method of claim 1 , wherein said EPO is non-glycosylated.
6. The method of claim 1 , wherein at least one chorionic gonadotrophin carboxy terminal peptide is glycosylated.
7. The method of claim 1 , wherein the amino acid sequence of said CTP-modified EPO polypeptide is set forth in SEQ ID NO: 51.
8. The method of claim 1 , wherein the amino acid sequence of said CTP-modified EPO polypeptide is set forth in SEQ ID NO: 3.
9. The method of claim 1 , wherein the amino acid sequence of said signal peptide comprises the sequence set forth in SEQ ID NO: 19.
10. The method of claim 1 , wherein said anemia comprises tumor-associated anemia, aplastic anemia, autoimmune hemolytic anemia, bone marrow transplantation, Churg-Strauss syndrome, Diamond Blackfan anemia, Fanconi's anemia, Felty syndrome, graft versus host disease, hematopoietic stem cell transplantation, hemolytic uremic syndrome, myelodysplasic syndrome, nocturnal paroxysmal hemoglobinuria, osteomyelofibrosis, pancytopenia, pure red-cell aplasia, purpura Schoenlein-Henoch, sideroblastic anemia, refractory anemia with excess of blasts, rheumatoid arthritis, Shwachman syndrome, sickle cell disease, thalassemia major, thalassemia minor, or thrombocytopenic purpura.
11. A method of stimulating erythropoiesis, said method comprising administering to a subject a therapeutically effective amount of a polypeptide consisting of a chorionic gonadotropin carboxy terminal peptide (CTP) modified erythropoietin (EPO), said CTP-modified EPO consisting of one chorionic gonadotropin carboxy terminal peptide (CTP) attached to the amino terminus of said EPO, and two chorionic gonadotropin CTPs attached to the carboxy terminus of said EPO, and wherein the amino acid sequence of said CTP-modified EPO consists of the sequence set forth in SEQ ID NO: 51, or wherein said CTP-modified EPO polypeptide consist of a chorionic gonadotropin carboxy terminal peptide (CTP) modified erythropoietin (EPO), said CTP-modified EPO polypeptide consisting of one chorionic gonadotropin carboxy terminal peptide (CTP) attached to the amino terminus of said EPO, and two chorionic gonadotropin CTPs attached to the carboxy terminus of said EPO, and a signal peptide attached to the amino terminal CTP, and wherein the amino acid sequence of said CTP-modified EPO consists of the sequence set forth in SEQ ID NO: 3.
12. The method of claim 11 , wherein the amino acid sequence of said signal peptide comprises the sequence set forth in SEQ ID NO: 19.
13. The method of claim 11 , wherein said EPO is glycosylated.
14. The method of claim 11 , wherein said EPO is non-glycosylated.
15. The method of claim 11 , wherein at least one chorionic gonadotrophin carboxy terminal peptide is glycosylated.