Variant form of urate oxidase and use thereof

• US 8,293,228 B2
• Filed: 04/20/2012
• Issued: 10/23/2012
• Est. Priority Date: 04/11/2005
• Status: Active Grant

• Current Assignee: Horizon Therapeutics USA, Inc. (Amgen Inc.)
• Original Assignee: Savient Pharmaceuticals Incorporated (Amgen Inc.)
• Inventors: Hartman, Jacob, Mendelovitz, Simona
• Primary Examiner(s): NASHED, NASHAAT T
• Application Number:
  US13/452,151
• Publication Number:
  US 20120225046A1
• Time in Patent Office: 186 Days
• Field of Search: None
• US Class Current: 424/94.3
• 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 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/06 Antigout agents, e.g. antihyperuricemic or uricosuric agents
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• 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: C12Y 107/03003 Factor-independent urate hydroxylase (1.7.3.3), i.e. uricase

Abstract

The present invention relates to genetically modified proteins with uricolytic activity. More specifically, the invention relates to proteins comprising truncated urate oxidases and methods for producing them, including PEGylated proteins comprising truncated urate oxidases.

Claims

1. A pharmaceutical composition comprising:

(a) an isolated uricase comprising an amino acid sequence of SEQ ID NO. 7; and

(b) a pharmaceutically acceptable carrier;

wherein the uricase is conjugated with a polymer.

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

4. The pharmaceutical composition of any one of claims 1 and 2, wherein the polymer is selected from the group consisting of poly(ethylene glycol), dextran, polypropylene glycol, hydroxypropylmethyl cellulose, carboxymethylcellulose, polyvinyl pyrrolidone and polyvinyl alcohol.

5. The pharmaceutical composition of claim 4 comprising 2-12 polyethylene glycol moieties on each uricase subunit.

6. The pharmaceutical composition of claim 5 comprising 3-10 polyethylene glycol moieties on each uricase subunit.

7. The pharmaceutical composition of claim 4, wherein each polyethylene glycol moiety has a molecular weight of about 1 kD and 100 kD.

8. The pharmaceutical composition of claim 7, wherein each polyethylene glycol moiety has a molecular weight of about 1 kD and 50 kD.

9. The pharmaceutical composition of claim 8, wherein each polyethylene glycol moiety has a molecular weight of about 5 kD and 20 kD.

10. The pharmaceutical composition of claim 9, wherein each polyethylene glycol moiety has a molecular weight of about 10 kD.

2. A pharmaceutical composition comprising:

(a) a uricase comprising an amino acid sequence of SEQ ID NO. 8; and

(c) a pharmaceutically acceptable carrier

(b) wherein the uricase is conjugated with a polymer.

11. A method for preparing a pharmaceutical composition for reducing uric acid levels in a biological fluid of a subject in need thereof, comprising:

(a) preparing a conjugate of the uricase of SEQ ID NO;

7 with a polymer; and

(b) adding the uricase conjugate to a pharmaceutically acceptable carrier.

13. The method of claim 11, wherein the biological fluid is blood.

15. The method of claim 11, wherein the polymer is selected from the group consisting of poly(ethylene glycol), dextran, polypropylene glycol, hydroxypropylmethyl cellulose, carboxymethylcellulose, polyvinyl pyrrolidone and polyvinyl alcohol.

12. A method for preparing a pharmaceutical composition for reducing uric acid levels in a biological fluid of a subject in need thereof, comprising:

(a) preparing a conjugate of the uricase of SEQ ID NO;

8 with a polymer; and

(b) adding the uricase conjugate to a pharmaceutically acceptable carrier.

14. The method of claim 12, wherein the biological fluid is blood.

16. The method of claim 12, wherein the polymer is selected from the group consisting of poly(ethylene glycol), dextran, polypropylene glycol, hydroxypropylmethyl cellulose, carboxymethylcellulose, polyvinyl pyrrolidone and polyvinyl alcohol.