PEG-urate oxidase conjugates and use thereof

• US 9,885,024 B2
• Filed: 07/22/2015
• Issued: 02/06/2018
• Est. Priority Date: 08/06/1998
• Status: Expired due to Fees

• Current Assignee: Horizon Therapeutics USA, Inc. (Amgen Inc.), Duke University
• Original Assignee: Horizon Pharma Rheumatology LLC (Amgen Inc.), Duke University
• Inventors: Williams, L. David, Hershfield, Michael S., Kelly, Susan J., Saifer, Mark G. P., Sherman, Merry R.
• Primary Examiner(s): Saidha, Tekchand
• Application Number:
  US14/806,494
• Publication Number:
  US 20160160188A1
• Time in Patent Office: 930 Days
• Field of Search: 424 944
• 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. A method for lowering elevated uric acid levels in a body fluid or tissue of a mammal, comprising the step of administering to said mammal an effective uric acid-lowering amount of a PEGylated uricase (PEG-uricase), said PEG-uricase comprising a purified uricase comprising at least two subunits in which each subunit is covalently linked to up to 12 strands of PEG, wherein each molecule of PEG has a molecular weight between about 5 kDa and 100 kDa wherein the PEG-uricase retains at least about 75% of the uricolytic activity of unconjugated uricase.

2. The method of claim 1, wherein each subunit of the PEG-uricase is covalently linked to an average of 2 to 10 strands of PEG.

3. The method of claim 1, wherein the administering step is selected from the group consisting of injections by intravenous, intradermal, subcutaneous, intramuscular and intraperitoneal routes or inhalation of an aerosolized formulation.

4. The method of claim 1, wherein said elevated uric acid levels are associated with a condition selected from the group consisting of gout, tophi, renal insufficiency, organ transplantation and malignant disease.

5. The method of claim 1, wherein said effective uric acid-lowering amount of PEG-uricase is administered at approximately weekly intervals.

6. The method of claim 1, wherein the uricase of the PEG-uricase is a mammalian uricase.

7. The method of claim 6, wherein the uricase of the PEG-uricase is a porcine uricase, a bovine uricase or an ovine uricase, or a combination thereof.

8. The method of claim 1, wherein the uricase of the PEG-uricase is recombinant.

9. The method of claim 8, wherein the uricase of the PEG-uricase comprises an amino acid sequence of porcine, bovine, ovine or baboon uricase, or a combination thereof.

10. The method of claim 1, wherein the uricase of the PEG-uricase is chimeric.

11. The method of claim 10, wherein the chimeric uricase of the PEG-uricase contains portions of porcine and baboon uricase.

12. The method of claim 1, wherein the uricase of the PEG-uricase is a fungal or microbial uricase.

13. The method of claim 12, wherein the fungal or microbial uricase is isolated from Aspergillus flavus, Arthrobacter globiformis or Candida utilis, or is a recombinant enzyme having substantially the sequence of one of those uricases.

14. The method of claim 1, wherein greater than 90% of the uricase of the PEG-uricase is in tetrameric form.

15. The method of claim 1, wherein the PEG of the PEG-uricase has an average molecular weight between about 10 kDa and 60 kDa.

16. The method of claim 1, wherein the PEG-uricase is in a pharmaceutical composition comprising a pharmaceutically acceptable carrier.

17. The method of claim 16, wherein said pharmaceutical composition is stabilized by lyophilization and dissolves upon reconstitution to provide solutions suitable for parenteral administration.

18. The method of claim 1, wherein the PEG-uricase is a conjugate of uricase that retains at least about 75% of the uricolytic activity of unconjugated uricase and is substantially reduced in immunogenicity, comprising a purified uricase containing no more than about 10% non-tetrameric aggregated uricase, said purified uricase comprising subunits in which each subunit of the uricase is covalently linked to an average of 2 to 10 strands of PEG, wherein each molecule of PEG has a molecular weight between about 5 kDa and 100 kDa.

19. The method of claim 1, wherein the PEG-uricase comprises 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.

20. The method of claim 1, wherein the PEG-uricase is a conjugate of uricase that retains uricolytic activity and has a prolonged half-life in vivo compared to an unconjugated uricase, said conjugate comprising a purified recombinant mammalian uricase conjugated to PEG, wherein greater than 90% of said uricase is in a tetrameric form, and wherein said PEG has an average molecular weight of about 10 kDa to about 60 kDa.

21. The method of claim 1, wherein the PEG-uricase is a conjugate of uricase that retains uricolytic activity and has a prolonged half-life in vivo compared to an unconjugated uricase, said conjugate comprising a purified recombinant uricase conjugated to PEG, wherein greater than 90% of said uricase is in a tetrameric form, wherein said uricase contains a portion of at least one of a porcine, bovine, ovine or baboon liver uricase, and wherein said PEG has an average molecular weight of about 10 kDa to about 60 kDa.