Long-acting polypeptides and methods of producing same

• US 9,896,494 B2
• Filed: 11/21/2014
• Issued: 02/20/2018
• Est. Priority Date: 02/03/2006
• Status: Active Grant

• Current Assignee: Opko Biologics Ltd. (Opko Health, Inc.)
• Original Assignee: Opko Biologics Ltd. (Opko Health, Inc.)
• Inventors: Fares, Fuad, Fima, Udi Eyal
• Primary Examiner(s): Kemmerer, Elizabeth C
• Assistant Examiner(s): Deberry, Regina M
• Application Number:
  US14/550,346
• Publication Number:
  US 20150152155A1
• Time in Patent Office: 1,187 Days
• Field of Search: None
• CPC Class Codes: A61K 38/19 Cytokines; Lymphokines; Interferons
• CPC Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/505 Erythropoietin [EPO]
• CPC Class Codes: C07K 14/555 Interferons [IFN]
• CPC Class Codes: C07K 14/59 Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g.hCG [human chorionic gonadotropin]; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
• CPC Class Codes: C07K 14/61 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/31 fusions, other than Fc, for prolonged plasma life, e.g. albumin
• CPC Class Codes: C07K 2319/35 containing a fusion for enhanced stability/folding during expression, e.g. fusions with chaperones or thioredoxin
• CPC Class Codes: C07K 2319/91 containing a motif for glycosylation

Abstract

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.

Claims

1. A method of extending C_max of a cytokine comprising the step of attaching one chorionic gonadotropin carboxy terminal peptide (CTP) to the amino terminus of said cytokine and two chorionic gonadotropin carboxy terminal peptides to the carboxy terminus of said cytokine, thereby extending C_max of said cytokine.

2. The method 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 method of claim 1, wherein said at least one CTP is glycosylated.

4. The method of claim 1, wherein said at least one CTP is truncated.

5. The method of claim 1, wherein at least one CTP is optionally attached to said cytokine via a linker.

6. The method of claim 5, wherein the linker is a peptide bond.

7. The method of claim 1, wherein said attaching produces a CTP-modified cytokine polypeptide, wherein when said polypeptide is a precursor polypeptide said polypeptide consists of a cytokine attached to a single chorionic gonadotropin carboxy terminal peptide (CTP) at the amino terminus of said cytokine, and two chorionic gonadotropin carboxy terminal peptides (CTP) attached to the carboxy terminus of said cytokine, and a signal peptide attached to the amino terminus of said one CTP, and when said polypeptide is a mature polypeptide, said polypeptide consists of a cytokine attached to a single chorionic gonadotropin carboxy terminal peptide (CTP) at the amino terminus of said cytokine, and two chorionic gonadotropin carboxy terminal peptides (CTP) attached to the carboxy terminus of said cytokine and said polypeptide lacks a signal peptide.

8. A pharmaceutical composition comprising a buffer, a tonicity agent, and a polypeptide consisting of a cytokine attached to a single chorionic gonadotropin carboxy terminal peptide (CTP) at the amino terminus of said cytokine, and two chorionic gonadotropin carboxy terminal peptides (CTP) attached to the carboxy terminus of said cytokine.

9. The pharmaceutical composition of claim 8, wherein said buffer is citrate.

10. The pharmaceutical composition of claim 8, wherein said tonicity agent is NaCl.

11. The pharmaceutical composition of claim 8, wherein said composition is at a pH of about 4-7.2.

12. The pharmaceutical composition 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.

13. The pharmaceutical composition of claim 8, wherein said at least one CTP is glycosylated.

14. The pharmaceutical composition of claim 8, wherein said at least one CTP is truncated.

15. The pharmaceutical composition of claim 8, wherein at least one CTP is optionally attached to said cytokine via a linker.

16. The pharmaceutical composition of claim 15, wherein the linker is a peptide bond.

17. The pharmaceutical composition of claim 8, wherein when said polypeptide is a precursor polypeptide, said polypeptide consists of a cytokine attached to a single chorionic gonadotropin carboxy terminal peptide (CTP) at the amino terminus of said cytokine, and two chorionic gonadotropin carboxy terminal peptides (CTP) attached to the carboxy terminus of said cytokine, and a signal peptide attached to the amino terminus of said one CTP, and when said polypeptide is a mature polypeptide, said polypeptide consists of a cytokine attached to a single chorionic gonadotropin carboxy terminal peptide (CTP) at the amino terminus of said cytokine, and two chorionic gonadotropin carboxy terminal peptides (CTP) attached to the carboxy terminus of said cytokine and said polypeptide lacks a signal peptide.

18. The pharmaceutical composition of claim 8, wherein said composition is a liquid.

19. The pharmaceutical composition of claim 8, wherein said composition is administered once weekly or once bi-weekly.

20. The pharmaceutical composition of claim 8, wherein said composition is formulated in a form suitable for an intramuscular (IM) injection, a subcutaneous (SC) injection, or an intravenous (IV) injection.