Methods For Inhibiting Viral Replication In Vivo

Abstract

The present invention is directed to methods of modulating viral replication in vivo comprising administering to an individual a therapeutically or prophylactically effective amount of a composition comprising arginine deiminase modified with polyethylene glycol, to methods of concurrently modulating viral replication and treating cancer, and to methods of modulating nitric oxide levels in a patient, among others.

1 . A method of inhibiting replication of human immunodeficiency virus in an individual comprising administering to said individual a composition comprising an arginine deiminase bonded to polyethylene glycol in an amount effective to inhibit human immunodeficiency virus replication in said individual.  
     
     
         2 . The method of  claim 1  further comprising the step of administering to said individual one or more compounds selected from the group consisting of antibiotics, anti-virals, antifungals, and anti-protozoan drugs.  
     
     
         3 . The method of  claim 1  further comprising the step of administering to said individual one or more other anti-viral compounds.  
     
     
         4 . The method of  claim 3  wherein said anti-viral compounds are one or more of azidovudine (AZT), didanosine (dideoxyinosine, ddl), d4T, zalcitabine (dideoxycytosine, ddC), nevirapine, lamivudine (epivir, 3TC), saquinavir (Invirase), ritonavir (Norvir), indinavir (Crixivan), delavirdine (Rescriptor), pegylated (PEG) interferon-α (IFN), or ribavirin.  
     
     
         5 . The method of  claim 1  wherein said composition is administered intramuscularly, intradermally, or intraperitoneally.  
     
     
         6 . The method of  claim 1  wherein said composition comprising an arginine deiminase bonded to polyethylene glycol is effective at a concentration of less than about 1 mM to inhibit human immunodeficiency virus replication by at least 50% in greater than 50% of cells in an assay to measure human immunodeficiency virus replication.  
     
     
         7 . The method of  claim 1  wherein the amount of arginine deiminase bonded to polyethylene glycol effective to inhibit human immunodeficiency virus replication is between about 40 IU/m and about 160 IU/m 2  per week.  
     
     
         8 . The method of  claim 1  wherein the amount of arginine deiminase bonded to polyethylene glycol effective to inhibit human immunodeficiency virus replication is about 160 IU/m2 per week.  
     
     
         9 . The method of  claim 1  wherein the amount of arginine deiminase bonded to polyethylene glycol effective to inhibit human immunodeficiency virus replication lowers plasma arginine levels to less than 5 μM.  
     
     
         10 . The method of  claim 1  wherein the arginine deiminase is covalently bonded via a linking group to polyethylene glycol, wherein each of said polyethylene glycol molecules has a molecular weight of about 10,000 to about 30,000.  
     
     
         11 . The method of  claim 1  wherein each of said polyethylene glycol molecules has a molecular weight of about 20,000.  
     
     
         12 . The method of  claim 10  wherein the linking group is selected from the group consisting of a succinimide group, an amide group, an imide group, a carbamate group, an ester group, an epoxy group, a carboxyl group, a hydroxyl group, a carbohydrate, a tyrosine group, a cysteine group, and a histidine group, and combinations thereof.  
     
     
         13 . The method of  claim 10  wherein the linking group is succinimidyl succinate.  
     
     
         14 . The method of  claim 1  wherein from about 7 to about 15 polyethylene glycol molecules are bonded to arginine deiminase.  
     
     
         15 . The method of  claim 1  wherein from about 9 to about 12 said polyethylene glycol molecules are bonded to arginine deiminase.  
     
     
         16 . The method of  claim 1  wherein said arginine deiminase is derived from a microorganism of the genus  Mycoplasma.    
     
     
         17 . The method of  claim 16  wherein said microorganism is selected from the group consisting of  Mycoplasma arginini, Mycoplasma hominus, Mycoplasma arthritides  and combinations thereof.  
     
     
         18 . The method of  claim 1  wherein the arginine deiminase has an amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 13, 14, 15, 16, 17, 18, 19, 20 or 21.  
     
     
         19 . The method of  claim 1  wherein the arginine deiminase has an amino acid sequence of SEQ ID NO: 1 or 4.  
     
     
         20 - 22 . (canceled)  
     
     
         23 . A method of treating an individual who is suspected of having been exposed to human immunodeficiency virus comprising the step of administering to said individual an amount of a composition comprising an arginine deiminase bonded to polyethylene glycol effective to inhibit human immunodeficiency virus replication in said individual.  
     
     
         24 . A method of inhibiting human immunodeficiency virus replication in an individual at risk for one or more viruses comprising administering to said individual an amount of a composition comprising an arginine deiminase bonded to polyethylene glycol effective to inhibit human immunodeficiency virus replication in said individual.  
     
     
         25 . A method of inhibiting human immunodeficiency virus replication in an individual who has been identified as having been infected with one or more viruses comprising administering to said individual an amount of a composition comprising an arginine deiminase bonded to polyethylene glycol effective to inhibit human immunodeficiency virus replication in said individual.  
     
     
         26 - 51 . (canceled)