SOLUBLE HYALURONIDASE GLYCOPROTEIN (sHASEGP), PROCESS FOR PREPARING THE SAME, USES AND PHARMACEUTICAL COMPOSITIONS COMPRISING THEREOF

Abstract

Provided are soluble neutral active Hyaluronidase Glycoproteins (sHASEGP's), methods of manufacture, and their use to facilitate administration for in vitro fertilization, as well as administration of 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.

What is claimed: 
     
         1 . An isolated soluble human PH20 glycoprotein, comprising an N-linked oligosaccharide, wherein the soluble human PH20 glycoprotein is catalytically active at neutral pH. 
     
     
         2 . The soluble PH20 glycoprotein of  claim 1  that comprises a plurality of N-linked oligosaccharides. 
     
     
         3 . The soluble PH20 glycoprotein of  claim 2 , wherein the N-linked oligosaccharides occur at two or more of N82, N166, N235, N254, N368, N391, and N393. 
     
     
         4 . The soluble human PH20 glycoprotein of  claim 3  that comprises the catalytic domain of PH20 and terminates, with reference to SEQ ID NO: 1, at residue 477, 478, 479, 480, 481, 482, or 483. 
     
     
         5 . The soluble human PH20 glycoprotein of  claim 1 , wherein the polypeptide is modified with a polymer. 
     
     
         6 . The soluble human PH20 glycoprotein of  claim 5 , wherein the polymer is dextran or PEG. 
     
     
         7 . The soluble human PH20 glycoprotein of  claim 5  that is PEGylated. 
     
     
         8 . The soluble human PH20 glycoprotein of  claim 1  that is super-sialylated. 
     
     
         9 . A composition, comprising the soluble human PH20 glycoprotein of  claim 1 . 
     
     
         10 . The composition of  claim 9  that is a pharmaceutical composition comprising the soluble human PH20 glycoprotein. 
     
     
         11 . The composition of  claim 9 , further comprising another pharmaceutically active agent. 
     
     
         12 . The composition of  claim 11 , 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 trichomonacidal agent, an anti-Parkinson agent, an anti-malarial agent, an anticonvulsant agent, an anti-depressant agent, an anti-arthritic agent, an anti-fungal agent, an antihypertensive agent, an antipyretic agent, an anti-parasite 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, a cytokine, an insulin, a sleep inducer and an antibody or antigen-binding fragment thereof. 
     
     
         13 . The composition of  claim 10  that is formulated for subcutaneous administration. 
     
     
         14 . The composition of  claim 11  that is formulated for subcutaneous administration. 
     
     
         15 . A method for increasing the diffusion of a therapeutic substance smaller than about 0.5 microns in diameter in a subject, comprising administering to a subject a soluble human glycoprotein of  claim 1  in an amount sufficient to open or to form channels smaller than about 0.5 microns in diameter and a therapeutic substance, whereby the diffusion of the therapeutic substance is increased.