Details for Patent: 8,765,685

Title:	Methods for treating edema by administering a Soluble Hyaluronidase Glycoprotein (sHASEGP)
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.
Inventor(s):	Bookbinder; Louis (San Diego, CA), Kundu; Anirban (San Diego, CA), Frost; Gregory I. (Del Mar, CA)
Assignee:	Halozyme, Inc. (San Diego, CA)
Filing Date:	May 16, 2012
Application Number:	13/506,783
Claims:	1. A method for treating a subject having edema, comprising administering a therapeutic amount of a hyaluronidase glycoprotein to the subject to treat the edema, whereby the edema is treated, wherein: the hyaluronidase glycoprotein is administered locally to the site of the edema or is administered systemically; the hyaluronidase glycoprotein: a) contains at least one sugar moiety that is covalently attached to an asparagine residue of the hyaluronidase polypeptide; b) is neutral active; and c) consists of either: i) a sequence of amino acids contained within SEQ ID NO:1 that includes amino acids 36-464 of SEQ ID NO: 1 and is truncated within the C-terminal region of SEQ ID NO:1 so that it does not comprise the full-length of the polypeptide whose sequence is set forth in 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 ii) a sequence of amino acids that contains amino acid substitutions in the sequence of amino acids of i), whereby the amino-acid substituted hyaluronidase glycoprotein consists of a sequence that has at least 91% amino acid sequence identity with a sequence of amino acids of i); or iii) a sequence of amino acids that has more than 99% sequence identity with a sequence of amino acids of i); or iv) a sequence of amino acids that at least 98% sequence identity with a sequence of amino acids of i) in which the C-terminus terminates at residue 483. 2. The method of claim 1, wherein the edema is interstitial edema, edema associated with retention of water and wastes associated with fat overloading of lipocytes, edema following head trauma or edema associated with brain tumors. 3. The method of claim 1, wherein the hyaluronidase glycoprotein is administered systemically. 4. The method of claim 1, wherein the glycoprotein is administered locally. 5. The method of claim 4, wherein the glycoprotein is administered locally in a slow release delivery vehicle. 6. The method of claim 5, wherein the slow release delivery vehicle is a colloidal dispersion system or polymer stabilized crystals. 7. The method of claim 6, wherein the vehicle is a colloidal dispersion system selected from among nanocapsules, microspheres, beads, lipid-based systems and oil-in-water emulsions. 8. The method of claim 7, wherein the vehicle is an oil-in-water emulsion selected from among micelles, mixed micelles and liposomes. 9. The method of claim 5, wherein the slow release delivery vehicle is selected from among biodegradable hydrogel matrices, dendritic polymer conjugates and multivesicular liposomes. 10. The method of claim 1, wherein 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, respectively. 11. The method of claim 1, wherein the hyaluronidase glycoprotein consists of a sequence of amino acids that has at least 98% amino acid sequence identity to a sequence of amino acids set forth as amino acid residues 36-483 of SEQ ID NO:1. 12. 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. 13. 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, respectively. 14. The method of claim 1, wherein the glycoprotein is conjugated to a polymer. 15. The method of claim 14, wherein the polymer is a polyethylene glycol (PEG) or a dextran. 16. The method of claim 3, wherein the glycoprotein is conjugated to a polymer. 17. The method of claim 16, wherein the polymer is a polyethylene glycol (PEG) or a dextran. 18. A method for treating a subject having edema, comprising administering a pharmaceutical composition comprising a therapeutic amount of a hyaluronidase glycoprotein to the subject to treat the edema, whereby the edema is treated, wherein: the hyaluronidase glycoprotein is administered locally to the site of the edema or is administered systemically; and the glycoprotein is produced in vitro in and secreted from a mammalian cell by a method comprising: a) 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; b) culturing the mammalian cell under conditions whereby the glycoprotein is expressed and secreted by the cell; and c) recovering from the cell a composition comprising the expressed and secreted hyaluronidase glycoprotein that is encoded by 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. 19. The method of claim 1, wherein a composition comprising the hyaluronidase glycoprotein that consists of the sequence of amino acids set forth as amino acids 36-482 of SEQ ID NO:1 is administered. 20. The method of claim 1, wherein the hyaluronidase glycoprotein is a polypeptide that has at least 98% sequence identity to the polypeptide whose sequence consists of residues 36-483 of SEQ ID NO: 1. 21. The method of claim 3, wherein the hyaluronidase glycoprotein is PEGylated. 22. The method of claim 1, wherein the hyaluronidase polypeptide is administered topically, intraarticularly, intracisternally, intraocularly, intraventricularly, intrathecally, intravenously, intramuscularly, intraperitoneally, intradermally or intratracheally. 23. The method of claim 1, wherein the subject has a tumor.