Variant forms of urate oxidase and use thereof

• US 9,926,538 B2
• Filed: 07/13/2017
• Issued: 03/27/2018
• 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:
  US15/649,488
• Publication Number:
  US 20170313995A1
• Time in Patent Office: 257 Days
• Field of Search: 4353201, 435191, 43525233, 435188, 4352523, 435 89, 4352542, 435325, 435348, 435 691, 435 9, 435 911, 424 944, 424 943, 530350, 536 232, 536 231
• 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 polypeptide comprising the amino acid sequence of SEQ ID NO:

8.

2. The polypeptide of claim 1 conjugated to polyethylene glycol (PEG).

3. The polypeptide of claim 1, wherein the PEG is monomethoxy PEG (mPEG).

4. The polypeptide of claim 3, wherein the mPEG has a molecular weight between 5 kDa and 20 kDa.

5. The polypeptide of claim 4, wherein the mPEG has a molecular weight of about 10 kDa.

6. The polypeptide of claim 5, wherein the mPEG is covalently, linked to a lysine residue of the polypeptide.

7. The polypeptide of claim 5, comprising 2-12 mPEG molecules attached to said polypeptide.

8. A conjugate comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:

8 linked to polyethylene glycol (PEG).

9. The conjugate of claim 8, wherein the PEG is monomethoxy PEG (mPEG).

10. The conjugate of claim 9, wherein the mPEG has a molecular weight between 5 kDa and 20 kDa.

11. The conjugate of claim 10, wherein the mPEG has a molecular weight of about 10 kDa.

12. The conjugate of claim 11, wherein 2-12 mPEG molecules are linked to the polypeptide.

13. A pharmaceutical composition comprising i) a conjugate comprising a polypeptide having the amino acid sequence of SEQ ID NO:

8 linked to polyethylene glycol (PEG); and

ii) phosphate buffered saline.

14. The pharmaceutical composition of claim 13, wherein the PEG is monomethoxy PEG (mPEG).

15. The pharmaceutical composition of claim 14, wherein the mPEG has a molecular weight between 5 kDa and 20 kDa.

16. The pharmaceutical composition of claim 15, wherein the mPEG has a molecular weight of about 10 kDa.

17. The pharmaceutical composition of claim 16, wherein 2-12 mPEG molecules are linked to the polypeptide.

18. The pharmaceutical composition of claim 17, comprising 8 mg of the polypeptide.

19. The pharmaceutical composition of claim 18, wherein said conjugate consists of the amino acid sequence of SEQ ID NO:

8 linked to PEG.

20. A uricase comprising the amino acid sequence of SEQ ID NO:

8.

21. The uricase of claim 20, wherein the uricase comprises a tetramer.

22. The uricase of claim 20, conjugated to polyethylene glycol (PEG).

23. The uricase of claim 22, wherein the PEG is monomethoxy PEG (mPEG).

24. The uricase of claim 23, wherein the mPEG has a molecular weight between 5 kDa and 20 kDa.

25. The uricase of claim 24, wherein the mPEG has a molecular weight of about 10 kDa.

26. The unease of claim 25, wherein the mPEG is covalently attached to a lysine residue of the unease.

27. The unease of claim 23, comprising 2-12 mPEG molecules per unease monomer.

28. The unease of claim 22, wherein the unease is formulated in a pharmaceutical composition comprising phosphate buffered saline.

29. The uricase of claim 28, wherein the pharmaceutical composition comprises 8 mg of the uricase.