Method of preserving infectious recombinant viruses, aqueous viral suspension, and use as medicament

Abstract

The present invention provides a novel method for preserving infectious recombinant viruses in frozen or liquid form, in which infectious viruses are preserved in an aqueous solution. The recombinant virus suspension comprises an aqueous sucrose solution at a concentration of 0.75 M or above, preferably between 0.75 M and 1.5 M, or more preferably at a concentration of 1 M. The preserved aqueous viral suspension further provides a medicament that can be used therapeutically or prophylactically for the treatment of a human or animal body by gene therapy.

1 . A method for the treatment of a human or animal body comprising administering a therapeutic or prophylactic amount of an aqueous suspension of infectious recombinant viruses to a human or animal in need of such treatment, wherein the infectious recombinant viruses are preserved by suspending the viruses in an aqueous sucrose solution at a concentration of above 0.75 M wherein the preserved recombinant viruses remain infectious.  
     
     
         2 . An aqueous suspension of infectious recombinant viruses comprising an aqueous sucrose solution at a concentration of above 0.75 M, wherein the pH of the aqueous solution is between 8 and 9.  
     
     
         3 . The aqueous suspension of  claim 2 , wherein the viruses are adenoviruses or retroviruses.  
     
     
         4 . The aqueous suspension of  claim 2 , wherein the aqueous solution is a buffer solution.  
     
     
         5 . The aqueous suspension of  claim 4 , wherein the buffer is selected from the group consisting of Tris-HCl, triethanolamine, diethanolamine, borate/HCl, glycine/NaOH, EPPS (N-(2-hydroxyethyly)piperazine-N′-(3-propanesulfonic acid)), bicine, TAPS (N-Tris-hydroxymethyl)methyl-3-aminopropanesulfonic acid) and tricine buffer.  
     
     
         6 . The aqueous suspension of  claim 2 , wherein the aqueous solution comprises at least one salt of a divalent cation selected from the group consisting of MgCl 2 , CaCl 2  and MnCl 2 .  
     
     
         7 . The aqueous suspension of  claim 6 , wherein the salt of the divalent cation is present in the aqueous solution at a concentration of between 0.1 mM and 5 mM.  
     
     
         8 . The aqueous suspension of  claim 2 , wherein the aqueous solution comprises 10 mM Tris-HCl buffer, 1 mM MgCl 2  and 1 M sucrose at a pH of approximately 8.5.  
     
     
         9 . The aqueous suspension of  claim 2 , wherein the aqueous solution comprises at least one stabilizer selected from the group consisting of monovalent salts, amino acids, and surfactants.  
     
     
         10 . The aqueous suspension of  claim 9 , wherein the monovalent salt is NaCl or KCl.  
     
     
         11 . The aqueous suspension of  claim 10 , wherein the monovalent salt is present at a concentration of between 0.05 and 1 M.  
     
     
         12 . The aqueous suspension of  claim 9 , wherein the surfactant is polysorbate 80 or nonaethylene glycol octyl phenol ether.  
     
     
         13 . The aqueous suspension of  claim 12 , wherein polysorbate 80 is present in the aqueous solution at a concentration of between 0.001 to 0.5% by weight.  
     
     
         14 . The aqueous suspension of  claim 2 , comprising 10 6  to 10 13  pfu/ml infectious recombinant viruses.  
     
     
         15 . The aqueous suspension of  claim 14 , wherein the aqueous solution comprises 10 mM Tris-HCl buffer, 1 mM MgCl 2 , 150 mM NaCl, 0.05% of polysorbate 80 and 1 M sucrose at a pH of approximately 8.5.  
     
     
         16 . A pharmaceutical composition comprising the aqueous suspension of infectious recombinant viruses of  claim 2 , in association with a pharmaceutically acceptable vehicle.  
     
     
         17 . The method of  claim 1 , wherein the pH of the aqueous solution is approximately 8.5.  
     
     
         18 . A method of administering infectious recombinant viruses to a human or animal body comprising administering an aqueous suspension of infectious recombinant viruses to said human or animal, wherein the recombinant viruses are preserved by suspending the viruses in an aqueous sucrose solution at a concentration of above 0.75 M and wherein the preserved recombinant viruses remain infectious.