Variant forms of urate oxidase and use thereof

• US 8,541,205 B2
• Filed: 05/01/2012
• Issued: 09/24/2013
• 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): Ha, Julie
• Application Number:
  US13/461,170
• Publication Number:
  US 20120309085A1
• Time in Patent Office: 511 Days
• Field of Search: None
• US Class Current: 435/91.1
• 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. An isolated nucleic acid comprising a nucleic acid sequence which encodes (a) 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 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;

(b) the uricase of (a) further comprising an amino terminal amino acid, wherein the amino terminal amino acid is alanine, glycine, proline, serine, or threonine;

(c) the uricase of (b) wherein the amino terminal amino acid is threonine;

(d) the uricase of (a) consisting of the amino acid sequence of SEQ ID NO. 8;

(e) the uricase of (a) wherein the N-terminal amino acid is methionine;

or

(f) the uricase of (e) comprising the amino acid sequence of SEQ ID NO. 7.

2. The isolated nucleic acid of claim 1, wherein the nucleic acid sequence is operatively linked to a heterologous promoter.

3. The nucleic acid of claim 2, wherein the promoter is the osmB promoter.

4. A nucleic acid vector comprising the nucleic acid of claim 2.

5. A host cell comprising the vector of claim 4.

6. An isolated nucleic acid comprising a nucleic acid sequence encoding a uricase comprising the amino acid sequence of SEQ ID NO. 7 or SEQ ID NO. 8.

7. The isolated nucleic acid of claim 6, wherein the nucleic acid sequence comprises SEQ ID NO. 9 or SEQ ID NO. 10.

8. The isolated nucleic acid of claim 6, wherein the nucleic acid sequence is operatively linked to a heterologous promoter.

9. The nucleic acid of claim 8, wherein the promoter is the osmB promoter.