Soluble hyaluronidase glycoprotein (sHASEGP), process for preparing the same, uses and pharmaceutical compositions comprising thereof

• US 20090214505A1
• Filed: 04/16/2009
• Published: 08/27/2009
• Est. Priority Date: 03/05/2003
• Status: Active Grant

• Current Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.)
• Original Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.)
• Inventors: Frost, Gregory I., Kundu, Anirban, Bookbinder, Louis H.
• Granted Patent: US 8,431,124 B2
• Time in Patent Office: Days
• US Class Current: 424/94.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• 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 38/47 acting on glycosyl compounds (3.2), e.g. cellulases, lactases
• CPC Class Codes: A61K 39/395 Antibodies agglutinins A61K38/36 ; as drug carriers A61K47/50; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
• CPC Class Codes: A61K 9/0048 Eye, e.g. artificial tears
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• 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 25/24 Antidepressants
• 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 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 33/00 Antiparasitic agents
• 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 7/04 : Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: A61P 7/10 : Antioedematous agents; Diuretics
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: C12N 9/2474 : Hyaluronoglucosaminidase (3.2.1.35), i.e. hyaluronidase
• CPC Class Codes: C12Y 302/01035 : 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

Provided are soluble neutral active Hyaluronidase Glycoproteins (sHASEGP'"'"'s), methods of manufacture, and their use to facilitate administration of other molecules or to alleviate glycosaminoglycan associated pathologies. Minimally active polypeptide domains of the soluble, neutral active sHASEGP domains are described that include asparagine-linked sugar moieties required for a functional neutral active hyaluronidase domain. Included are modified amino-terminal leader peptides that enhance secretion of sHASEGP. Sialated and pegylated forms of the sHASEGPs also are provided. Methods of treatment by administering sHASEGPs and modified forms thereof also are provided.

Claims

1. A method for treating a subject having an excess of a substrate for a hyaluronidase, comprising:

administering an amount of a soluble hyaluronidase glycoprotein (sHASEGP) sufficient to reduce that amount of the substrate to effect treatment, wherein;

the sHASEGP is a soluble neutral active glycoprotein that contains at least one sugar moiety that is covalently attached to an asparagine residue; and

the sHASEGP contains a sequence of amino acids included in SEQ ID NO;

1 or a sequence that has at least about 91% amino acid sequence identity with a sequence of amino acids included in SEQ ID NO;

1, whereby the sHASEGP is soluble, N-glycosylated and neutral active.

2. The method of claim 1, wherein the substrate is associated with a herniated disk.

3. The method of claims 1, wherein the sHASEGP is encoded by a nucleic acid molecule that encodes amino acids 36-482 of SEQ ID NO:

1 or amino acids 1-482 of SEQ ID NO;

1.

4. The method of claim 3, wherein the nucleic acid molecule has a sequence of nucleotides set forth in SEQ ID NO:

48.

5. The method of claim 1, wherein the sHASEGP includes a sequence of amino acids set forth in SEQ ID NO:

1 that is truncated at an amino acid residue that is or is between amino acid residues 467 to 483.

6. The method of claim 1, wherein the sHASEGP includes a sequence of amino acids set forth in SEQ ID NO:

1 that is truncated at an amino acid residue selected from among 467, 477, 478, 479, 480, 481, 482 and 483.

7. The method of claim 6, wherein the sHASEGP is secreted in CHO cells.

8. The method of claim 1, wherein the sHASEGP has the sequence of amino acids set forth as amino acids 36-477, 36-478, 36-479, 36-480, 36-481, 36-482, or 36-483 of SEQ ID NO:

1.

9. The method of claim 1, wherein the sHASEGP is modified with a polymer.

10. The method of claim 9, wherein the polymer is a polyethylene glycol (PEG) or a dextran.

11. The method of claim 8, wherein the sHASEGP is modified with a polymer.

12. The method of claim 11, wherein the polymer is a polyethylene glycol (PEG) or a dextran.

13. The method of claim 1, wherein the excess substrate is produced from a scar tissue.

14. The method of claim 13, wherein the scar tissue is a glial scar resulting from spinal cord injury, is scar tissue resulting from surgery or is a keloid scar.

15. The method of claim 2, wherein the sHASEGP includes a sequence of amino acids set forth in SEQ ID NO:

1 that is truncated at an amino acid residue that is or is between amino acid residues 467 to 483.

16. The method of claim 2, wherein the sHASEGP includes a sequence of amino acids set forth in SEQ ID NO:

1 that is truncated at an amino acid residue selected from among 467, 477, 478, 479, 480, 481, 482 and 483.

17. The method of claim 16, wherein the sHASEGP is secreted in CHO cells.

18. The method of claim 2, wherein the sHASEGP has the sequence of amino acids set forth as amino acids 36-477, 36-478, 36-479, 36-480, 36-481, 36-482, or 36-483 of SEQ ID NO:

1.

19. A method for treating a herniated disk by chemonucleolysis, comprising contacting the herniated disk with an amount of a soluble hyaluronidase glycoprotein (sHASEGP) sufficient to effect chemonucleolysis of the hernia, wherein the sHASEGP is a soluble neutral active glycoprotein that contains at least one sugar moiety that is covalently attached to an asparagine residue;

and

the sHASEGP contains a sequence of amino acids included in SEQ ID NO;

1 or a sequence that has at least about 91% amino acid sequence identity with a sequence of amino acids included in SEQ ID NO;

1, whereby the sHASEGP is soluble, N-glycosylated and neutral active.