Stable formulations of a hyaluronan-degrading enzyme

• US 9,993,529 B2
• Filed: 06/15/2012
• Issued: 06/12/2018
• Est. Priority Date: 06/17/2011
• Status: Active Grant

• Current Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.)
• Original Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.)
• Inventors: Yang, Tzung-Horng, Vaughn, Daniel Edward, Labarre, Michael James, Caster, Christopher L., Nicol, Francois, Kim, Donghyun
• Primary Examiner(s): Mondesi, Robert B
• Assistant Examiner(s): Epstein, Todd M
• Application Number:
  US13/507,262
• Publication Number:
  US 20130022588A1
• Time in Patent Office: 2,188 Days
• Field of Search: None
• CPC Class Codes: A61K 38/28 Insulins
• CPC Class Codes: A61K 38/47 acting on glycosyl compounds (3.2), e.g. cellulases, lactases
• CPC Class Codes: A61K 47/02 Inorganic compounds
• 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: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07K 14/62 Insulins
• CPC Class Codes: C07K 5/06 Dipeptides
• CPC Class Codes: C07K 5/06086 with the first amino acid being basic
• CPC Class Codes: C12N 9/2402 hydrolysing O- and S- glycosyl compounds (3.2.1)
• CPC Class Codes: C12Y 302/01035 Hyaluronoglucosaminidase (3.2.1.35), i.e. hyaluronidase

Abstract

Provided are compositions that are stable formulations of a hyaluronan-degrading enzyme or are stable co-formulations of a fast-acting insulin and a hyaluronan degrading enzyme, including a recombinant human PH20 (rHuPH20).

Claims

1. A composition, comprising:

a therapeutically effective amount of a soluble PH20 polypeptide, wherein the soluble PH20 polypeptide is active at neutral pH; and

lysyl lysine (Lys-Lys) at a concentration that is about 10 mM to about 200 mM, inclusive, whereby the soluble PH20 polypeptide in the composition, when incubated at a temperature of 37°

C. to 42°

C., inclusive, exhibits greater hyaluronidase activity than a composition that is identical, except that the composition does not contain the Lys-Lys.

2. The composition of claim 1, wherein the concentration of soluble PH20 polypeptide is at least 10 U/mL.

3. The composition of claim 1, wherein the Lys-Lys concentration is 50 mM to 150 mM, inclusive.

4. The composition of claim 1, wherein the amount of a soluble PH20 polypeptide is between 10 U/mL to 5000 U/mL, inclusive.

5. The composition of claim 1, wherein the pH of the composition is between about 6.5 to 8.0, inclusive.

6. The composition of claim 1, comprising a stabilizing agent, an antioxidant, a tonicity modifier to maintain the osmolality at between about 245 mOsm/kg to 500 mOsm/kg, inclusive, or a buffering agent.

7. The composition of claim 6, wherein the composition comprises a stabilizing agent and the stabilizing agent is selected from among an amino acid, an amino acid derivative, an amine, a sugar, a polyol, a salt and a surfactant.

8. The composition of claim 5, that includes a stabilizing agent selected from among an amino acid, an amino acid derivative, an amine, a sugar, a polyol, a salt and a surfactant.

9. The composition of claim 7, wherein the stabilizing agent is a surfactant and the amount of surfactant, as a % of mass concentration (w/v) in the formulation, is between 0.0005% to 1.0%, inclusive.

10. The composition of claim 9, wherein the surfactant is selected from among a polypropylene glycol, polyethylene glycol, glycerin, sorbitol, poloxamer and polysorbate.

11. The composition of claim 6, wherein the composition comprises an antioxidant.

12. The composition of claim 11, wherein the antioxidant is selected from among cysteine, tryptophan and methionine.

13. The composition of claim 12, wherein the antioxidant is methionine.

14. The composition of claim 11, wherein the antioxidant is at a concentration from between 5 mM to 50 mM, inclusive.

15. The composition of claim 3, comprising a tonicity modifier to maintain the osmolality of between 245 mOsm/kg to 500 mOsm/kg, inclusive.

16. The composition of claim 6, wherein the composition comprises a tonicity modifier and the tonicity modifier is selected from among glycerin, salt, amino acids, polyalcohols and trehalose.

17. The composition of claim 15, wherein the tonicity modifier is NaCl, wherein the concentration of NaCl is between about 20 mM to 200 mM, inclusive.

18. The composition of claim 16, wherein the tonicity modifier is NaCl at a concentration between about 20 mM to 200 mM, inclusive.

19. The composition of claim 18, wherein the concentration of NaCl is less than 150 mM.

20. The composition of claim 6, wherein the composition comprises a buffering agent and the buffering agent is selected from among Tris, histidine, phosphate and citrate.

21. The composition of claim 20, wherein the buffering agent is a phosphate that is sodium phosphate.

22. The composition claim 20, wherein the concentration of the buffering agent is between 1 mM to 100 mM, inclusive.

23. The composition of claim 1, wherein the soluble PH20 polypeptide is selected from ovine PH20, bovine PH20 and a soluble human PH20, wherein the soluble human PH20 lacks all or a portion of the glycosylphosphatidylinositol (GPI) anchor attachment sequence.

24. The composition of claim 23, wherein the soluble PH20 polypeptide is a soluble human PH20 that consists of the sequence of amino acids set forth in any of SEQ ID NOS:

4-9, 47-48, 234-246, 248-254, and 267-273, or a sequence of amino acids that exhibits at least 95% sequence identity to any of SEQ ID NOS;

4-9, 47-48, 234-246, 248-254, and 267-273.

25. The composition of claim 23, wherein the soluble PH20 polypeptide is a soluble human PH20 that consists of the sequence of amino acids set forth in any of SEQ ID NOS:

4-9.

26. The composition of claim 1, wherein the concentration of Lys-Lys is 10 mM to 100 mM, inclusive.

27. The composition of claim 1, wherein:

the pH of the composition is between 6.5 to 7.2, inclusive;

and the composition comprises;

a soluble PH20 polypeptide at a concentration that is between about 100 U/mL to 500 U/mL, inclusive;

NaCl at a concentration less than 140 mM NaCl;

a surfactant that is polysorbate 80 at a percentage (%) of mass concentration (w/v) of between 0.01% to 0.05%, inclusive;

methionine at a concentration that is between 5 mM to 20 mM, inclusive; and

sodium phosphate at a concentration that is between 5 mM to 50 mM, inclusive.

28. The composition of claim 27, wherein the soluble PH20 polypeptide is selected from among ovine PH20, bovine PH20 and a soluble human PH20, wherein the soluble human PH20 lacks all or a portion of the glycosylphosphatidylinositol (GPI) anchor attachment sequence.

29. The composition of claim 1, wherein the soluble PH20 polypeptide in the composition, when incubated at a temperature of 38°

C. to 42°

C., exhibits at least 50% more hyaluronidase activity than a composition that is identical, except that the composition does not contain the Lys-Lys.

30. The composition of claim 1, further comprising a fast-acting insulin.

31. The composition of claim 30, wherein the concentration of Lys-Lys is 30 mM to 120 mM, inclusive.

32. The composition of claim 30, wherein the fast-acting insulin is a fast-acting insulin analog.

33. The composition of claim 32, wherein the insulin analog is selected from among insulin aspart, insulin lispro and insulin glulisine.

34. The composition of claim 33, wherein the soluble PH20 polypeptide is selected from among ovine PH20, bovine PH20 and a soluble human PH20, wherein the soluble human PH20 lacks all or a portion of the glycosylphosphatidylinositol (GPI) anchor attachment sequence.

35. The composition of claim 30, wherein the concentration of fast-acting insulin is between 10 U/mL to 1000 U/mL, inclusive.

36. The composition of claim 30, wherein:

the fast-acting insulin is an insulin analog that is glulisine and the concentration of Lys-Lys is 50 to 105 mM;

or

the fast-acting insulin is an insulin analog that is insulin aspart or insulin lispro and the concentration of Lys-Lys is 80 to 100 mM.

37. The composition of claim 30 that is formulated as a single dosage or as a multiple dosage for direct administration, wherein if the composition is for multiple dosage administration, the composition comprises an anti-microbially effective amount of a preservative or mixture of preservatives.

38. The composition of claim 37, wherein:

the composition is formulated for multiple dosage administration; and

the preservative(s) in the formulation comprises one or more of a phenolic preservative(s), a non-phenolic preservative(s) or a phenolic preservative(s) and a non-phenolic preservative(s).

39. The composition of claim 38, wherein the preservative(s) is(are) selected from among phenol, m-cresol, methylparaben, benzyl alcohol, thimerosal, benzalkonium chloride, 4-chloro-1-butanol, chlorhexidine dihydrochloride, chlorhexidine digluconate, L-phenylalanine, EDTA, bronopol, phenylmercuric acetate, glycerol, imidurea, chlorhexidine, sodium dehydroacetate, o-cresol, p-cresol, chlorocresol, cetrimide, benzethonium chloride, ethylparaben, propylparaben, butylparaben and any combinations thereof.

40. The composition of claim 39, wherein the preservative(s) is(are) phenol, m-cresol or phenol and m-cresol.

41. The composition of claim 40, wherein the preservatives are phenol and m-cresol and the amount as a % of mass concentration (w/v) in the formulation is between 0.1% to 0.25% phenol and between 0.05% to 0.2% m-cresol, inclusive.

42. The composition of claim 38, wherein the total amount of the one or more preservative agents as a percentage (%) of mass concentration (w/v) in the formulation is or is between 0.1% and 0.4%, inclusive.

43. The composition of claim 30, wherein:

the pH of the composition is between 6.8 to 7.4, inclusive; and

the composition comprises;

a soluble PH20 polypeptide in an amount between 100 U/mL to 1000 U/mL, inclusive;

a fast-acting insulin analog that is insulin glulisine is an amount between 10 U/mL to 1000 U/mL, inclusive;

Lys-Lys at a concentration between 50 mM to 105 mM, inclusive;

NaCl at a concentration of less than 100 mM;

a surfactant that is polysorbate 20 at a percentage (%) of mass concentration (w/v) of between 0.0005% to 0.005%, inclusive;

methionine at a concentration between 5 mM to 20 mM, inclusive; and

a preservative(s) that comprises phenol at a percentage (%) of mass concentration (w/v) of between 0.1% to 0.25% and m-cresol at a % w/v of between 0.05% to 0.2%, each inclusive.

44. The composition of claim 30, wherein:

the pH of the composition is between 6.8 to 7.4, inclusive; and

the composition comprises;

a soluble PH20 polypeptide in an amount between 100 U/mL to 1000 U/mL, inclusive;

a fast-acting insulin analog that is insulin aspart or insulin lispro is an amount between 10 U/mL to 1000 U/mL, inclusive;

Lys-Lys at a concentration between 80 mM to 100 mM, inclusive;

NaCl at a concentration of less than 30 mM;

a surfactant that is polysorbate 20 at a percentage (%) of mass concentration (w/v) of between 0.0005% to 0.005%, inclusive;

methionine at a concentration between 5 mM to 20 mM, inclusive; and

a preservative(s) that comprises phenol at a percentage (%) of mass concentration (w/v) of between 0.1% to 0.25% and m-cresol at a % w/v of 0.05% to 0.2%, each inclusive.

45. A syringe or vial, comprising the composition of claim 1.

46. A syringe or vial, comprising the composition of claim 30.

47. A closed loop system, comprising the composition of claim 37.

48. An insulin pump, comprising the composition of claim 37.

49. An insulin pen, comprising the composition of claim 30.

55. A method of reducing a symptom of diabetes in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the composition of claim 30.

50. A composition, comprising:

a soluble PH20 polypeptide at a concentration of between 10 U/mL to 5000 U/mL, inclusive, wherein the soluble PH20 polypeptide is active at neutral pH;

lysyl lysine (Lys-Lys) at a concentration of about 10 mM to 200 mM, inclusive, wherein the pH of the composition is between about 6.5 to 8.0, inclusive.

51. The composition of claim 50, comprising NaCl at a concentration between about 20 mM to 200 mM, inclusive.

52. The composition of claim 51, comprising lysyl lysine (Lys-Lys) at a concentration of about 50 mM to 150 mM, inclusive.

53. The composition of claim 50, wherein the soluble PH20 polypeptide is a soluble human PH20 that consists of the sequence of amino acids set forth in any of SEQ ID NOS:

4-9, 47-48, 234-246, 248-254, and 267-273, or a sequence of amino acids that exhibits at least 95% sequence identity to any of SEQ ID NOS;

4-9, 47-48, 234-246, 248-254, and 267-273.

54. The composition of claim 50, further comprising a fast-acting insulin.