Methods for treating a disease characterized by an excess of hyaluronan by administering a soluble hyaluronidase glycoprotein (sHASEGP)

• US 8,431,124 B2
• Filed: 04/16/2009
• Issued: 04/30/2013
• Est. Priority Date: 03/05/2003
• Status: Active Grant

• Current Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.)
• Original Assignee: Halozyme, Inc. (Halozyme Therapeutics, Inc.)
• Inventors: Bookbinder, Louis H., Kundu, Anirban, Frost, Gregory I.
• Primary Examiner(s): Kosson, Rosanne
• Application Number:
  US12/386,473
• Publication Number:
  US 20090214505A1
• Time in Patent Office: 1,475 Days
• Field of Search: 424/94.62, 514/21.2, 435/201, 435/69.1
• US Class Current: 424/94.62
• 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 a disease or condition in which there is an excess of a substrate for a hyaluronidase compared to in the absence of the disease or condition, comprising:

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

an excess of substrate is an increased accumulation of the substrate in a diseased tissue or tissue manifesting the condition compared to a normal tissue;

the glycoprotein contains at least one sugar moiety that is covalently attached to an asparagine residue of the hyaluronidase polypeptide;

is neutral active; and

the glycoprotein consists of 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 or contains amino acid substitutions in 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, whereby the amino-acid substituted hyaluronidase glycoprotein consists of a sequence that has at least 95% amino acid sequence identity with 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.

2. The method of claim 1, wherein:

the substrate for the hyaluronidase is produced or accumulates in the lumbar spine, thereby causing intradiscal pressure of a herniated disk; and

reducing the amount of the substrate effects treatment of the herniated disk.

3. The method of claim 1, wherein the glycoprotein is produced in and secreted from a mammalian cell by a method comprising:

introducing an expression vector comprising a polynucleotide that consists of the sequence of nucleotides set forth as nucleotides 106-1446 of SEQ ID NO;

6 or degenerates thereof or the sequence of nucleotides set forth in SEQ ID NO;

48 or degenerates thereof inserted between flanking sequences in the vector into a mammalian cell;

culturing the mammalian cell under conditions whereby the glycoprotein is expressed and secreted by the cell; and

recovering a composition comprising the expressed and secreted hyaluronidase glycoprotein.

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

48.

5. The method of claim 1, wherein the glycoprotein consists of the sequence of amino acids 36-477, 36-478, 36-479, 36-480, 36-481, 36-482, or 36-483 of SEQ ID NO:

1.

6. The method of claim 1, wherein the glycoprotein contains amino acid substitutions in 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, whereby the amino-acid substituted hyaluronidase glycoprotein consists of a sequence that has at least 95% amino acid sequence identity with 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.

7. The method of claim 3, wherein the mammalian cell is a CHO cell.

8. The method of claim 1, wherein a composition comprising the glycoprotein that consists of the sequence of amino acids set forth as amino acids 36-482 of SEQ ID NO:

1 is administered.

9. The method of claim 1, wherein the glycoprotein is conjugated to 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 glycoprotein is conjugated to 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 glycoprotein consists of the sequence of amino acids 36-477, 36-478, 36-479, 36-480, 36-481, 36-482, or 36-483 of SEQ ID NO:

1.

16. The method of claim 2, wherein the glycoprotein contains amino acid substitutions in 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, whereby the amino-acid substituted hyaluronidase glycoprotein consists of a sequence that has at least 95% amino acid sequence identity with 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.

17. The method of claim 2, wherein the glycoprotein is produced in and secreted from a mammalian cell by a method comprising:

introducing an expression vector comprising a polynucleotide that consists of the sequence of nucleotides set forth as nucleotides 106-1446 of SEQ ID NO;

6 or degenerates thereof or the sequence of nucleotides set forth in SEQ ID NO;

48 or degenerates thereof inserted between flanking sequences in the vector into a mammalian cell;

culturing the mammalian cell under conditions whereby the glycoprotein is expressed and secreted by the cell; and

recovering a composition comprising the expressed and secreted hyaluronidase glycoprotein.

18. The method of claim 2, wherein a composition comprising the glycoprotein that consists of the sequence of amino acids set forth as amino acids 36-482 of SEQ ID NO:

1 is administered.

19. A method for treating a herniated disk by chemonucleolysis, comprising contacting the herniated disk with an amount of a hyaluronidase glycoprotein sufficient to effect chemonucleolysis of the hernia, wherein:

the glycoprotein contains at least one sugar moiety that is covalently attached to an asparagine residue of the hyaluronidase polypeptide;

is neutral active; and

the glycoprotein consists of 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 or contains amino acid substitutions in 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, whereby the amino-acid substituted hyaluronidase glycoprotein consists of a sequence that has at least 95% amino acid sequence identity with 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.

20. A method for treating a subject having a disease or condition characterized by an excess of a substrate for a hyaluronidase, comprising:

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

an excess of substrate is an increased accumulation of the substrate in a diseased tissue or tissue manifesting the condition compared to a normal tissue;

the glycoprotein contains at least one sugar moiety that is covalently attached to an asparagine residue of the hyaluronidase polypeptide;

is neutral active; and

the glycoprotein;

a) consists of a C-terminally truncated polypeptide of SEQ ID NO;

1 that includes at least amino acids 36-464 of SEQ ID NO;

1, wherein the truncation is at a residue selected from among 477, 478, 479, 480, 481, 482 and 483 of SEQ ID NO;

1;

or

b) the glycoprotein contains amino acid substitutions in the polypeptide of a), whereby the amino-acid substituted hyaluronidase glycoprotein consists of a sequence that has at least 95% amino acid sequence identity with the polypeptide of a);

or

c) the glycoprotein has more than 99% sequence identity of with the polypeptide of a);

or

d) the glycoprotein has more than 97% sequence identity with the polypeptide of a), wherein truncation is at residue 483.

21. The method of claim 20, wherein the disease or condition is a scar or herniated disk.