Soluble hyaluronidase glycoprotein (sHASEGP), process for preparing the same, uses and pharmaceutical compositions comprising thereof
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.
1. A substantially purified glycoprotein, comprising a neutral active soluble hyaluronidase polypeptide and at least one N-linked sugar moiety, wherein:
the N-linked sugar moiety is covalently attached to an asparagine residue of the polypeptide;
the asparagine residue is selected from amino acids 82, 166, 235, 254, 368, 393 or 490 as set forth in SEQ ID NO: 1;
the glycoprotein contains at least residues 36-364 of SEQ ID NO:1; and
the glycoprotein contains a contiguous sequence of amino acid residues of SEQ ID NO:1, but it is C-terminally truncated so that it does not contain all or a portion of the GPI anchor, whereby the glycoprotein is soluble and neutral active.
2. The glycoprotein of claim 1 , wherein the polypeptide is modified with a polymer.
3. The glycoprotein of claim 2 , wherein the polymer is polyethylene glycol (PEG) or dextran.
4. A pharmaceutical composition, comprising the glycoprotein of claim 1 , and a pharmaceutically acceptable carrier.
5. A pharmaceutical composition, comprising the glycoprotein of claim 3 , and a pharmaceutically acceptable carrier.
6. The pharmaceutical composition of claim 4 , further comprising a pharmaceutically active agent.
7. The pharmaceutical composition of claim 5 , further comprising a pharmaceutically active agent.
8. The pharmaceutical composition of claim 6 , wherein the pharmaceutically active agent is selected from among a chemotherapeutic agent, an analgesic agent, an anti-inflammatory agent, an antimicrobial agent, an amoebicidal agent, a trichomonocidal agent, an anti-parkinson agent, an anti-malarial agent, an anti-convulsant agent, an anti-depressant agent, an antiarthritics agent, an anti-fungal agent, an antihypertensive agent, an antipyretic agent, an anti-parasitic agent, an antihistamine agent, an alpha-adrenergic agonist agent, an alpha blocker agent, an anesthetic agent, a bronchial dilator agent, a biocide agent, a bactericide agent, a bacteriostat agent, a beta adrenergic blocker agent, a calcium channel blocker agent, a cardiovascular drug agent, a contraceptive agent, a decongestant agent, a diuretic agent, a depressant agent, a diagnostic agent, an electrolyte agent, a hypnotic agent, a hormone agent, a hyperglycemic agent, a muscle relaxant agent, a muscle contractant agent, an ophthalmic agent, a parasympathomimetic agent, a sedative agent, a sympathomimetic agent, a tranquilizer agent, a viricide agent, a vitamin agent, a non-steroidal anti-inflammatory agent, an angiotensin converting enzyme inhibitor agent, and a sleep inducer.
9. The pharmaceutical composition of claim 6 , wherein the pharmaceutically active agent is an antibody.
10. The pharmaceutical composition of claim 7 , wherein the pharmaceutically active agent is an antibody.
11. A combination, comprising:
a first composition, comprising the glycoprotein of claim 1 ; and
a second composition, comprising a pharmaceutically active agent, wherein the first and second compositions are separate or combined to form a single composition.
12. The combination of claim 11 , wherein:
the glycoprotein is PEGylated; and
the pharmaceutically active agent is an antibody.
13. A method of treatment of a tumor in a subject, comprising administering the composition of claim 4 .
14. A method of treatment of a tumor in a subject, comprising administering the composition of claim 5 .
15. The pharmaceutical composition of claim 7 , wherein the pharmaceutically active agent is selected from among a chemotherapeutic agent, an analgesic agent, an anti-inflammatory agent, an antimicrobial agent, an amoebicidal agent, a trichomonocidal agent, an anti-parkinson agent, an anti-malarial agent, an anti-convulsant agent, an anti-depressant agent, an antiarthritics agent, an anti-fungal agent, an antihypertensive agent, an antipyretic agent, an anti-parasitic agent, an antihistamine agent, an alpha-adrenergic agonist agent, an alpha blocker agent, an anesthetic agent, a bronchial dilator agent, a biocide agent, a bactericide agent, a bacteriostat agent, a beta adrenergic blocker agent, a calcium channel blocker agent, a cardiovascular drug agent, a contraceptive agent, a decongestant agent, a diuretic agent, a depressant agent, a diagnostic agent, an electrolyte agent, a hypnotic agent, a hormone agent, a hyperglycemic agent, a muscle relaxant agent, a muscle contractant agent, an ophthalmic agent, a parasympathomimetic agent, a sedative agent, a sympathomimetic agent, a tranquilizer agent, a viricide agent, a vitamin agent, a non-steroidal anti-inflammatory agent, an angiotensin converting enzyme inhibitor agent, and a sleep inducer.
16. The glycoprotein of claim 1 , wherein the glycoprotein is PEGylated.
17. A pharmaceutical composition, comprising the glycoprotein of claim 16 .
18. The pharmaceutical composition of claim 17 , further comprising an antibody.
19. The combination of claim 12 , wherein the antibody is a monoclonal antibody.
20. The pharmaceutical composition of claim 18 , wherein the antibody is a monoclonal antibody.
21. The method of claim 14 , wherein the composition is administered subcutaneously.
22. The method of claim 14 , wherein the composition is administered intravenously.