Pharmaceutical preparation and method of treatment of human malignancies with arginine deprivation

Abstract

The present invention provides an isolated and substantially purified recombinant human arginase having sufficiently high enzymatic activity and stability to maintain Adequate Arginine Depletion in a patient. The present invention also provides a pharmaceutical composition comprising the modified invention enzyme and method for treatment of diseases using the pharmaceutical composition.

1 - 27 . (canceled) 
     
     
         28 : A method of treatment of human liver, breast, colon or rectal malignancies, comprising administering to a subject in need thereof a modified, full-length recombinant human arginase I polypeptide of 80-100% purity, which is covalently linked to at least one polyethylene glycol (PEG) molecule. 
     
     
         29 : The method of  claim 28 , wherein the modified, full-length recombinant human arginase I polypeptide has an extended half-life of at least 3 days. 
     
     
         30 : The method of  claim 28 , wherein the modified, full-length recombinant human arginase I polypeptide has an amino acid sequence encoded by a nucleic acid of SEQ ID NO. 2. 
     
     
         31 : The method of  claim 28 , wherein the modified, full-length recombinant human arginase I polypeptide has the amino acid sequence of SEQ ID NO. 3. 
     
     
         32 : The method of  claim 28 , wherein the wherein the modified, full-length recombinant human arginase I polypeptide has an extended half-life relative to the half-life of an unmodified full-length recombinant human arginase I. 
     
     
         33 : The method of  claim 28 , wherein the administration of the modified, full-length recombinant human arginase I polypeptide reduces the physiological arginine level in the subject to below 10 μM for at least 3 days. 
     
     
         34 : A method of treatment of human liver, breast, colon or rectal malignancies, comprising administering to a subject in need thereof a modified, full-length recombinant human arginase I polypeptide comprising the amino acid sequence of SEQ ID NO. 3 which is of 80-100% purity, covalently linked to at least one polyethylene glycol (PEG) molecule, and has an extended half-life of at least 3 days. 
     
     
         35 : The method of  claim 34 , wherein the administration of the modified, full-length recombinant human arginase I polypeptide reduces the physiological arginine level in the subject to below 10 μM for at least 3 days. 
     
     
         36 : A method of treatment of human liver, breast, colon or rectal malignancies, comprising administering to a subject in need thereof a modified, full-length recombinant human arginase I polypeptide, which is covalently linked to at least one polyethylene glycol (PEG) molecule, wherein the administration of the modified, full-length recombinant human arginase I polypeptide reduces the physiological arginine level in the subject to below 10 μM for at least 3 days.