PEG-urate oxidase conjugates and use thereof

• US 7,723,089 B2
• Filed: 04/19/2001
• Issued: 05/25/2010
• Est. Priority Date: 08/06/1998
• Status: Expired due to Fees

• Current Assignee: Horizon Therapeutics USA, Inc. (Amgen Inc.), Duke University
• Original Assignee: Duke University, Mountain View Pharmaceuticals, Inc.
• Inventors: Sherman, Merry R., Saifer, Mark G. P., Kelly, Susan J., Williams, L. David, Hershfield, Michael S.
• Primary Examiner(s): Saidha; Tekchand
• Application Number:
  US09/839,946
• Publication Number:
  US 20030166249A1
• Time in Patent Office: 3,323 Days
• Field of Search: 424/94.4, 435/189, 435/191, 435/252.3, 435/320.1, 435/190, 435/440, 530/350, 536/23.2
• US Class Current: 435/190
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• 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: C12N 9/0046 with oxygen as acceptor (1.7.3)
• CPC Class Codes: C12N 9/0048 Uricase (1.7.3.3)
• CPC Class Codes: C12N 9/0093 acting on CH or CH2 groups (1.17)
• CPC Class Codes: C12N 9/96 Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
• CPC Class Codes: C12Y 107/03003 Factor-independent urate hydroxylase (1.7.3.3), i.e. uricase

Abstract

A naturally occurring or recombinant urate oxidase (uricase) covalently coupled to poly(ethylene glycol) or poly(ethylene oxide) (both referred to as PEG), wherein an average of 2 to 10 strands of PEG are conjugated to each uricase subunit and the PEG has an average molecular weight between about 5 kDa and 100 kDa. The resulting PEG-uricase conjugates are substantially non-immunogenic and retain at least 75% of the uricolytic activity of the unmodified enzyme.

Claims

1. An isolated recombinant tetrameric mammalian uricase, wherein said uricase is in a substantially tetrameric form, wherein less than 10% of said uricase is in a non-tetrameric aggregated form, and wherein said uricase is chimeric.

2. The isolated tetrameric uricase of claim 1, wherein said chimeric uricase contains portions of porcine liver and baboon liver uricases.

3. The isolated tetrameric uricase of claim 2, wherein said chimeric uricase is pig-baboon chimeric uricease.

6. The isolated tetrameric uricase of claim 1, wherein said uricase comprises an amino terminus and a carboxyl terminus, and wherein the uricase is truncated at one terminus or both termini.

4. An isolated recombinant tetrameric mammalian uricase, wherein said uricase is recombinant porcine uricase containing lysine in place of arginine at residue number 291 in SEQ ID NO:

1 and serine in place of threonine at residue number 301 in SEQ ID NO;

1.

7. The isolated tetrameric uricase of claim 4, wherein said uricase comprises an amino terminus and a carboxyl terminus, and wherein the uricase is truncated at one terminus or both termini.

5. An isolated recombinant tetrameric mammalian uricase, wherein said uricase has the sequence of baboon liver uricase in which tyrosine 97 in SEQ ID NO:

2 has been replaced by histidine.

8. The isolated tetrameric uricase of claim 5 wherein said uricase comprises an amino terminus and a carboxyl terminus, and wherein the uricase is truncated at one terminus or both termini.