Neutral active soluble truncated PH20 polypeptides and uses thereof

• US 9,284,543 B2
• Filed: 02/21/2012
• Issued: 03/15/2016
• Est. Priority Date: 12/09/2008
• Status: Active Grant

• Current Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.), Zriha Hlavin Industries Ltd.
• Original Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.)
• Inventors: Wei, Ge, Selvam, Krishnasamy Panneer, Bookbinder, Louis, Frost, Gregory I.
• Primary Examiner(s): Swope, Sheridan
• Application Number:
  US13/385,527
• Publication Number:
  US 20120213767A1
• Time in Patent Office: 1,484 Days
• US Class Current: 1/1
• 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: A61K 47/61 the organic macromolecular compound being a polysaccharide or a derivative thereof
• CPC Class Codes: A61K 47/6435 the peptide or protein in the drug conjugate being a connective tissue peptide, e.g. collagen, fibronectin or gelatin
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/08 of the prostate
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 Ophthalmic 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 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 33/04 Amoebicides
• CPC Class Codes: A61P 33/06 Antimalarials
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: C12N 9/2408 : acting on alpha -1,4-glucosidic bonds
• CPC Class Codes: C12N 9/2474 : Hyaluronoglucosaminidase (3.2.1.35), i.e. hyaluronidase
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Soluble PH20 polypeptides are provided, including extended soluble PH20 polypeptides, and uses thereof. Also provided are other C-terminally truncated PH20 polypeptides and partially deglycosylated PH20 polypeptides and uses thereof.

Claims

1. A truncated soluble PH20 hyaluronidase, wherein:

the truncated PH20 hyaluronidase is neutral active;

is soluble; and

consists of any sequence of amino acid residues within SEQ NO;

107 that terminates at residue 469, 470 or 471 of SEQ ID NO;

107.

3. The truncated soluble PH20 hyaluronidase of claim 1 that is N-glycosylated.

4. The truncated soluble PH20 hyaluronidase of claim 3, wherein the N-glycosylated polypeptide comprises an N-acetylglucosamine moiety linked to each of at least three asparagine (N) residues selected from among amino acid residues N82, N166, N235, N254, N368 and N393 of SEQ ID NO:

107.

5. The truncated soluble PH20 hyaluronidase of claim 4, wherein the three asparagine residues are amino acid residues 235, 368 and 393 of SEQ ID NO:

107.

8. The truncated soluble PH20 hyaluronidase of claim 1 that is modified by modification selected from among sialation, albumination, farnesylation, carboxylation, hydroxylation and phosphorylation.

9. The truncated soluble PH20 hyaluronidase of claim 1 that is modified by a polymer.

10. The truncated soluble PH20 hyaluronidase of claim 9, wherein the polymer is dextran or PEG.

11. The truncated soluble PH20 hyaluronidase of claim 1 that is substantially purified.

12. A conjugate, comprising a conjugated moiety linked directly or via a linker to the C-terminus or N-terminus of the truncated soluble PH20 hyaluronidase of claim 1.

13. The conjugate of claim 12, wherein the truncated soluble PH20 hyaluronidase is conjugated to a moiety selected from among a multimerization domain, toxin, detectable label or drug.

14. The conjugate of claim 13, wherein the truncated soluble PH20 hyaluronidase is conjugated to an Fc domain.

15. A composition, comprising the truncated soluble PH20 hyaluronidase polypeptide of claim 1.

16. A composition, comprising a truncated soluble PH20 hyaluronidase polypeptide of claim 10.

21. A composition, comprising the truncated soluble PH20 hyaluronidase polypeptide of claim 1.

2. A truncated soluble PH20 hyaluronidase that is selected from among a polypeptide that contains an amino acid substitution or amino acid substitutions in the sequence of amino acids set forth as amino acids 36-469, 36-470 or 36-471 of SEQ ID NO:

107 that has at least 95% sequence identity with the sequence of amino acids set forth as amino acids 36-469, 36-470 or 36-471 of SEQ ID NO;

107, wherein;

the amino acid sequence of the amino acid-substituted polypeptide differs from the sequence of amino acids set forth as amino acids 36-469, 36-470 or 36-471 of SEQ ID NO;

107 only by amino acid substitutions;

the polypeptide is soluble; and

the polypeptide has hyaluronidase activity.

6. The truncated soluble PH20 hyaluronidase of claim 2 that is a polypeptide that contains an amino acid substitution or amino acid substitutions in the sequence of amino acids set forth as amino acids 36-469, 36-470 or 36-471 of SEQ ID NO:

107, whereby the amino acid-substituted polypeptide consists of a sequence of amino acids that has at least 98% sequence identity with the corresponding sequence of amino acids set forth as amino acids 36-469, 36-470 or 36-471 of SEQ ID NO;

107.

17. The truncated soluble PH20 hyaluronidase of claim 2 that is N-glycosylated.

18. The truncated soluble PH20 hyaluronidase of claim 17, wherein the N-glycosylated polypeptide comprises an N-acetylglucosamine moiety linked to each of at least three asparagine (N) residues selected from among amino acid residues N82, N166, N235, N254, N368 and N393 of SEQ ID NO:

107.

19. The truncated soluble PH20 hyaluronidase of claim 2 that is modified by a polymer.

20. The truncated soluble PH20 hyaluronidase of claim 19, wherein the polymer is dextran or PEG.

7. A truncated soluble PH20 hyaluronidase that consists of the sequence of amino acids set forth as amino acids 36-469, 36-470 or 36-471 of SEQ ID NO:

107.

22. A composition, comprising the truncated soluble PH20 hyaluronidase polypeptide of claim 7.