Variant forms of urate oxidase and use thereof

• US 9,670,467 B2
• Filed: 03/27/2015
• Issued: 06/06/2017
• Est. Priority Date: 04/11/2005
• Status: Active Grant

• Current Assignee: Horizon Therapeutics USA, Inc. (Amgen Inc.)
• Original Assignee: Horizon Pharma Rheumatology LLC (Amgen Inc.)
• Inventors: Hartman, Jacob, Mendelovitz, Simona
• Primary Examiner(s): Ha, Julie
• Application Number:
  US14/671,246
• Publication Number:
  US 20150197732A1
• Time in Patent Office: 802 Days
• Field of Search: 424 943, 424 944, 435188, 435191, 435 91, 435 89, 43525233, 4353201, 536 232, 530300, 530350
• CPC Class Codes: A61K 38/44 Oxidoreductases (1)
• 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: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 19/06 Antigout agents, e.g. antihyperuricemic or uricosuric agents
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 9/0046 with oxygen as acceptor (1.7.3)
• CPC Class Codes: C12N 9/0048 Uricase (1.7.3.3)
• CPC Class Codes: C12Y 107/03003 Factor-independent urate hydroxylase (1.7.3.3), i.e. uricase

Abstract

Genetically modified proteins with uricolytic activity are described. Proteins comprising truncated urate oxidases and methods for producing them, including PEGylated proteins comprising truncated urate oxidase are described.

Claims

1. A method of reducing uric acid levels in a subject in need thereof, comprising administering a polyethylene glycol-uricase conjugate to the subject wherein the polyethylene glycol-uricase conjugate comprises a uricase comprising a mammalian uricase amino acid sequence truncated at the amino terminus, or the carboxy terminus, or both the amino and carboxy termini by 1-6 amino acids, and further comprising an amino acid substitution with threonine at position 7 (D7T), an amino acid substitution with threonine at position 46 (S46T), an amino acid substitution with lysine at position 291 (R291K) and an amino acid substitution with serine at position 301 (T301S), said truncation and amino acid substitution being relative to a naturally occurring pig uricase having an amino acid sequence of SEQ ID NO:

11.

2. The method of claim 1, wherein the uric acid levels are reduced in the plasma of the subject.

3. The method of claim 1, wherein the uricase further comprises an amino terminal amino acid, wherein the amino terminal amino acid is alanine, glycine, proline, serine, or threonine.

4. The method of claim 3, wherein the amino terminal amino acid of the uricase is threonine.

5. The method of claim 4 wherein the amino acid sequence of the uricase comprises SEQ ID NO:

8.

6. The method of claim 1, wherein the uricase comprises porcine, bovine, ovine, or baboon liver uricase.

7. The method of claim 1, wherein each polyethylene glycol molecule has a molecular weight between 5 kD and 20 kD.

8. The method of claim 1, wherein each polyethylene glycol molecule has a molecular weight of about 10 kD.

9. The method of claim 1, wherein plasma uric acid levels are lowered to 6.8 mg/dl or less.

10. The method of claim 1, wherein plasma uric acid levels are lowered to 6.0 mg/dl or less.

11. The method of claim 1, wherein the subject has a plasma uric acid level of 6.0 mg/dl or less for at least 80% of a treatment period.

12. The method of claim 1, wherein the polyethylene glycol-uricase conjugate is administered every two weeks.

13. The method of claim 1, wherein the polyethylene glycol-uricase conjugate is administered at a dosage of 8 mg of uricase.

14. The method of claim 1, wherein the polyethylene glycol-uricase conjugate is administered at a dosage of 8 mg of uricase every 2 weeks.

15. The method of claim 1, wherein the polyethylene glycol-uricase conjugate is administered at a dosage of 8 mg of uricase every 4 weeks.

16. The method of claim 1, wherein the polyethylene glycol-uricase conjugate is administered by intravenous infusion.

17. The method of claim 1, wherein the polyethylene glycol-uricase conjugate is provided in a pharmaceutical composition comprising 8 milligrams of uricase per milliliter of solution.

18. The method of claim 1, wherein the polyethylene glycol-uricase conjugate comprising 8 milligrams of uricase is diluted into 250 milliliters of saline solution for infusion.

19. The method of claim 1, wherein the polyethylene glycol-uricase conjugate is administered over a 120 minute period.

20. The method of claim 1, wherein the polyethylene glycol-uricase conjugate is administered over a treatment period of 12 weeks, 24 weeks, 36 weeks 48 weeks, 3 months, 6 months, 8 months or 12 months.

21. The method of claim 1, further comprising administering at least one anti-inflammatory compound prior to administration of the polyethylene glycol-uricase conjugate.

22. The method of claim 21, wherein the at least one anti-inflammatory compound is selected from a corticosteroid, an antihistamine, and a Nonsteroidal Anti-Inflammatory Drug (NSAID).

23. The method of claim 22, wherein at least one corticosteroid, at least one antihistamine and at least one NSAID are administered.

24. The method of claim 22, wherein the corticosteroid is selected from betamethasone, budesonide, cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisone and triamcinolone.

25. The method of claim 24, wherein the corticosteroid is hydrocortisone and/or prednisone.

26. The method of claim 25, wherein hydrocortisone is administered at a dosage of 100 to 500 mg.

27. The method of claim 25, wherein prednisone is administered at a dosage of 5 to 50 mg.

28. The method of claim 22, wherein the NSAID is selected from ibuprofen, indomethacin, naproxen, aspirin, celecoxib and valdecoxib.

29. The method of claim 22, wherein the antihistamine is selected from azatadine, brompheniramine, cetirizine, chlorpheniramine, clemastine, cyproheptadine, desloratadine, dexchlorpheniramine, dimenhydrinate, diphenhydramine, doxylamine, fexofenadine, hydroxyzine, loratadine and phenindamine.

30. The method of claim 29, wherein the antihistamine is fexofenadine.

31. The method of claim 30, wherein the fexofenadine is administered at a dosage of 30 to 180 mg.

32. The method of claim 30, wherein the fexofenadine is administered 1 to 4 hours prior to intravenous infusion of the pharmaceutical composition.

33. The method of claim 1, further comprising administering acetaminophen prior to administration of the polyethylene glycol-uricase conjugate.

34. The method of claim 33, wherein the acetaminophen is administered at a dosage of 500 to 1500 mg.

35. The method of claim 33, wherein the acetaminophen is administered 1 to 4 hours prior to intravenous infusion of the pharmaceutical composition.

36. The method of claim 1, further comprising administering at least one corticosteroid, at least one antihistamine and acetaminophen prior to administration of the polyethylene glycol-uricase conjugate.

37. The method of claim 36, wherein the corticosteroid is hydrocortisone and/or prednisone, and the antihistamine is fexofenadine.

38. A method of reducing uric acid levels in a subject in need thereof, comprising administering a uricase to the subject, wherein the uricase comprises a mammalian uricase amino acid sequence truncated at the amino terminus, or the carboxy terminus, or both the amino and carboxy termini by 1-6 amino acids, and further comprises an amino acid substitution with threonine at position 7 (D7T), an amino acid substitution with threonine at position 46 (S46T), an amino acid substitution with lysine at position 291 (R291K) and an amino acid substitution with serine at position 301 (T301S), said truncation and amino acid substitution being relative to a naturally occurring pig uricase having an amino acid sequence of SEQ ID NO:

11.