Liquid formulation of long-acting human growth hormone conjugate

• US 9,061,072 B2
• Filed: 07/14/2011
• Issued: 06/23/2015
• Est. Priority Date: 07/14/2010
• Status: Active Grant

• Current Assignee: Hanmi Science Co., Ltd
• Original Assignee: Hanmi Science Co., Ltd
• Inventors: Hong, Sung Hee, Lee, Byung Sun, Lee, Jae Min, Bae, Sung Min, Kwon, Se Chang, Im, Dae Seong
• Primary Examiner(s): STOICA, ELLY GERALD
• Application Number:
  US13/809,783
• Publication Number:
  US 20130115231A1
• Time in Patent Office: 1,440 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 38/27 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: A61K 47/12 Carboxylic acids; Salts or anhydrides thereof
• CPC Class Codes: A61K 47/26 Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/68 the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/08 Solutions composition of solutions A61K47/00
• CPC Class Codes: A61P 5/06 of the anterior pituitary hormones, e.g. TSH, ACTH, FSH, LH, PRL, GH
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Abstract

Disclosed is a liquid formulation of long-acting human growth hormone (hGH) conjugate, free of albumin, which can guarantee the stability of the long-acting hGH conjugate when stored over a long period of time, wherein the long-acting human growth hormone conjugate includes a human growth hormone linked to an immunoglobulin Fc region, and has a prolonged in vivo stability compared to the native form. The liquid formulation of hGH conjugate including a pH 5.0˜6.0 buffer, a sugar alcohol, a salt and a non-ionic surfactant is free of human serum albumin and other hazardous factors which are potentially contaminated with viruses, and can provide excellent storage stability customized for a long-acting hGH conjugate composed of an hGH polypeptide and an immunoglobulin Fc region which has higher molecular weight and in vivo durability, compared to the native.

Claims

1. A liquid formulation of a long-acting human growth hormone conjugate, comprising

a pharmaceutically effective amount of the long-acting human growth hormone conjugate in which the human growth hormone and an immunoglobulin Fc region are linked to each other via a polyethylene glycol;

and

an albumin-free stabilizer, said stabilizer consisting essentially of a buffer of pH 5.0-6.0, a sugar alcohol, a polysorbate as a non-ionic surfactant and sodium chloride as a tonicity agent.

2. The liquid formulation of the long-acting human growth hormone conjugate of claim 1, wherein the sugar alcohol is selected from the group consisting of mannitol, sorbitol and a combination thereof.

3. The liquid formulation of the long-acting human growth hormone conjugate of claim 1, wherein the sugar alcohol is used in an amount of from 1 to 10% (w/v) based the total volume of the formulation.

4. The liquid formulation of the long-acting human growth hormone conjugate of claim 1, wherein the buffer is a citrate or phosphate buffer.

5. The liquid formulation of the long-acting human growth hormone conjugate of claim 1, wherein the buffer ranges in pH from 5.2 to 6.0.

6. The liquid formulation of the long-acting human growth hormone conjugate of claim 1, wherein the buffer ranges in pH from 5.2 to 5.5.

7. The liquid formulation of the long-acting human growth hormone conjugate of claim 1, wherein the sodium chloride is used in a concentration of from 5 to 200 mM.

8. The liquid formulation of the long-acting human growth hormone conjugate of claim 1, wherein the polysorbate is polysorbate 80.

9. The liquid formulation of the long-acting human growth hormone conjugate of claim 1, wherein the polysorbate is used in an amount of from 0.001 to 0.05% (w/v) based on the total volume of the formulation.

11. The liquid formulation of the long-acting human growth hormone conjugate of claim 1, wherein the human growth hormone and the immunoglobulin Fc region are linked to each other via a polyethylene glycol, and the stabilizer consisting essentially of a citrate buffer with a pH of 5.2 to 6.0, 1-10% (w/v) mannitol, 0.001-0.05% polysorbate 80, and 5-200 mM NaCl.

12. The liquid formulation of the long-acting human growth hormone conjugate of claim 1, wherein the human growth hormone has an amino acid sequence identical to that of a native human growth hormone.

13. The liquid formulation of the long-acting human growth hormone conjugate of claim 1, wherein the immunoglobulin Fc region is selected from the group consisting of an Fc region of IgG, IgA, IgD, IgE, and IgM.

14. The liquid formulation of the long-acting human growth hormone conjugate of claim 13, wherein the immunoglobulin Fc region is a dimer or multimer of a single stranded immunoglobulin having domains of the same origin.

15. The liquid formulation of the long-acting human growth hormone conjugate of claim 13, wherein the immunoglobulin Fc region is an IgG4 Fc region.

16. The liquid formulation of the long-acting human growth hormone conjugate of claim 15, wherein the immunoglobulin Fc region is an aglycosylated human IgG4 Fc region.

17. The liquid formulation of the long-acting human growth hormone conjugate of claim 1, wherein the long-acting human growth hormone conjugate comprises a human growth hormone and an immunoglobulin Fc region with a linkage via a non-peptidyl polymer.

18. The liquid formulation of the long-acting human growth hormone conjugate of claim 17, wherein the non-peptidyl polymer is selected from the group consisting of polyethylene glycol, polypropylene glycol, copolymers of ethylene glycol and propylene glycol, polyoxyethylated polyols, polyvinyl alcohol, polysaccharides, dextran, polyvinyl ethyl ether, polylactic acid, polylactic-glycolic acid, lipid polymers, chitins, hyaluronic acid, and a combination thereof.

19. A method for producing the liquid formulation of the long-acting human growth hormone conjugate as in one of claims 1-6, 7-13, and 14-18, comprising:

combining the long-acting human growth hormone conjugate with a stabilizer, said stabilizer consisting essentially of a buffer with a pH of 5.0-6.0, a sugar alcohol, a polysorbate as a non-ionic surfactant and sodium chloride as a tonicity agent to give the liquid formulation comprising the long-acting human growth hormone conjugate and the stabilizer.

20. A method for stabilizing a long-acting human growth hormone conjugate according to claim 1 in a liquid formulation comprising the long-acting human growth hormone conjugate, said method comprising:

combining the long-acting human growth hormone conjugate with a stabilizer, said stabilizer consisting essentially of a buffer with a pH of 5.0-6.0, a sugar alcohol, a polysorbate as a non-ionic surfactant and sodium chloride as a tonicity agent to obtain the liquid formulation comprising the long-acting human growth hormone conjugate and the stabilizer, said liquid formulation retains the activity of the long-acting human growth hormone conjugate at a level of 90% or greater of the original activity after storage at 40±

2°

C. for one weeks.

21. The liquid formulation of the long-acting human growth hormone conjugate of claim 1, wherein the polysorbate is used in an amount of from 0.001 to 0.01% (w/v) based on the total volume of the formulation.

10. A liquid formulation of a long-acting human growth hormone conjugate comprising

a pharmaceutically effective amount of the long-acting human growth hormone conjugate in which the human growth hormone and an immunoglobulin Fc region are linked to each other via a polyethylene glycol;

and

an albumin-free stabilizer, said stabilizer consisting essentially of a buffer of pH 5.0-6.0, a sugar alcohol, a polysorbate as a non-ionic surfactant, sodium chloride as a tonicity agent, and one or more ingredient selected from the group consisting of a sugar, a polyalcohol and an amino acid.