Adenovirus formulations

Abstract

The invention relates to viral formulations and related pharmaceutical products for use in gene therapy and/or vaccine applications. Especially preferred viral formulations disclosed herein are liquid adenovirus formulations, which show improved stability when stored in about the 2-8° C. range while also being compatible with parenteral administration. These formulations comprise a buffer, a sugar, a salt, a divalent cation, a non-ionic detergent, as well as a free radical scavenger and/or a chelating agent to inhibit free radical oxidation.

What is claimed is:  
     
         1 . A virus formulation comprising: 
 a) a purified virus    b) a buffer;    c) a sugar;    d) a salt;    e) a divalent cation; and,    f) a non-ionic detergent.    
     
     
         2 . A virus formulation of  claim 1  with an adenovirus concentration in the range from about 1×10 7  vp/mL to about 1×10 13  vp/mL and a total osmolarity in a range from about 200 mOs/L to about 800 mOs/L.  
     
     
         3 . A virus formulation of  claim 1  with a virus concentration in the range from about 1×10 7  vp/mL to about 1×10 13  vp/mL, wherein the buffer is selected from a group of buffers acceptable for human parenteral use, preferably a Tris buffer, at a pH from about 7.5 to about 8.5.  
     
     
         4 . A virus formulation of  claim 3  wherein the sugar is sucrose at a weight to volume percentage from about 2% to about 7.5% and the salt is sodium chloride from about 25 mM to about 250 mM, such that the total osmolarity of the formulation is a range from about 200 mOs/L to about 800 mOs/L.  
     
     
         5 . A virus formulation of  claim 4  wherein the divalent cation is selected from the group consisting of MgCl 2  and CaCl 2  in an amount from about 0.1 mM to about 5 mM.  
     
     
         6 . A virus formulation of  claim 5  wherein the non-ionic detergent is selected from the group consisting of Polysorbate-80 and Polysorbate-40 at a concentration range from about 0.001% to about 2%.  
     
     
         7 . A virus formulation of  claim 1  with a concentration in the range from about 1×10 7  vp/mL to about 1×10 13  vp/mL and a total osmolarity in a range from about 200 mOs/L to about 800 mOs/L which comprises a about 5.0 mM Tris, at pH 8.0; sucrose in a weight to volume range of about 5%; NaCl at about 75 mM, MgCl 2  at about 1 mM to 2 mM, and either Polysorbate-80 at a concentration of about 0.02% or Polysorbate40 at a concentration of about 0.005%.  
     
     
         8 . A virus formulation of  claim 7  which wherein the formulation is buffered with about 5.0 mM Tris-HCl, at pH 8.0; sucrose is present a about 5%, NaCl is present at about 75 mM, MgCl 2  at 1 mM, and Polysorbate-80 at 0.001% with the total osmolarity approximately 310 mOs/L.  
     
     
         9 . A virus formulation of  claim 2  comprising adenovirus and a formulation selected from the group consisting of formulation number A105, A110, A111, A126, A127, A128, A129, A130, A131, A155, A159, A160, A165, A167, A168, A169, A170, A171, A172 and A173.  
     
     
         10 . A virus formulation comprising: 
 a) a purified virus    b) a buffer;    c) a sugar;    d) a salt;    e) a divalent cation;    f) a non-ionic detergent; and,    g) an inhibitor of free radical oxidation.    
     
     
         11 . A virus formulation of  claim 10  with a concentration in the range from about 1×10 7  vp/mL to about 1×10 13  vp/mL and a total osmolarity in a range from about 200 mOs/L to about 800 mOs/L.  
     
     
         12 . A virus formulation of  claim 10  with a virus concentration in the range from about 1×10 7  vp/mL to about 1×10 13  vp/mL, wherein the buffer is selected from a group of buffers acceptable for human parenteral use, preferably a Tris buffer, at a pH from about 7.5 to about 8.5.  
     
     
         13 . A virus formulation of  claim 12  wherein the sugar is sucrose at a weight to volume percentage from about 2% to about 7.5% and the salt is sodium chloride from about 25 mM to about 250 mM, such that the total osmolarity of the formulation is a range from about 200 mOs/L to about 800 mOs/L.  
     
     
         14 . A virus formulation of  claim 13  wherein the divalent cation is selected from the group consisting of MgCl 2  and CaCl 2  in an amount from about 0.1 mM to about 5 mM.  
     
     
         15 . A virus formulation of  claim 14  wherein the non-ionic detergent is selected from the group consisting of Polysorbate-80 and Polysorbate40 at a concentration range from about 0.001% to about 2%.  
     
     
         16 . A virus formulation of  claim 15  wherein the inhibitor of free radical oxidation is selected from the group consisting of ethanol, EDTA, an EDTA/ethanol combination, triethanolamine, and sodium citrate.  
     
     
         17 . A virus formulation of  claim 10  with a concentration in the range from about 1×10 7  vp/mL to about 1×10 13  vp/mL and a total osmolarity in a range from about 200 mOs/L to about 800 mOs/L which comprises a about 5.0 mM Tris, at pH 8.0; sucrose in a weight to volume range of about 5%; NaCl at about 75 mM, MgCl 2  at about 1 mM to 2 mM, either Polysorbate-80 at a concentration of about 0.02% or Polysorbate-40 at a concentration of about 0.005%, EDTA is present at about 100 μM and ethanol at about 0.5%.  
     
     
         18 . A virus formulation of  claim 17  which comprises about 5.0 mM Tris-HCl, at pH 8.0; sucrose at about 5%, NaCl at about 75 mM, MgCl 2  from about 1 mM to 2 mM, Polysorbate-80 at about 0.005%, EDTA at about 100 μM and ethanol at about 0.5%.  
     
     
         19 . A virus formulation of  claim 11  comprising adenovirus and a formulation selected from the group consisting of formulation number A105, A110, A111, A112, A121, A126, A127, A128, A129, A130, A131, A155, A159, A160, A165, A167, A168, A169, A170, A171, A172 and A173.  
     
     
         20 . A virus formulation of  claim 2  which further comprises plasmid DNA at a concentration from about 0.01 mg/mL to about 10 mg/mL.  
     
     
         21 . A virus formulation of  claim 11  which further comprises plasmid DNA at a concentration from about 0.01 mg/mL to about 10 mg/mL.  
     
     
         22 . A formulation of  claim 20  which comprises 5.0 mM Tris-HCl, at pH 8.0; sucrose at about 5%, NaCl at about 75 mM, MgCl 2  from about 1 mM to about 2 mM, Polysorbate-80 at about 0.005%, and plasmid DNA at about 1 mg/mL.  
     
     
         23 . A virus formulation of  claim 21  which comprises 5.0 mM Tris-HCl, at pH 8.0; sucrose at about 5%, NaCl at about 75 mM, MgCl 2  from about 1 mM to about 2 mM, Polysorbate-80 at about 0.005%, EDTA at about 100 μM, ethanol at about 0.5% and plasmid DNA at about 1 mg/mL.  
     
     
         24 . A virus formulation comprising at least one inhibitor of free radical oxidation selected from the group consisting of ethanol, EDTA, an EDTA/ethanol combination, triethanolamine, and sodium citrate.  
     
     
         25 . A virus formulation of  claim 24  wherein a purified virus and the inhibitor(s) of free radical oxidation further comprise a buffer, a sugar, a salt, a divalent cation; and a non-ionic detergent.  
     
     
         26 . A virus formulation of  claim 25  with an adenovirus concentration in the range from about 1×10 7  vp/mL to about 1×10 13  vp/ml and a total osmolarity in a range from about 200 mOs/L to about 800 mOs/L.  
     
     
         27 . A virus formulation of  claim 26  with an adenovirus concentration in the range from about 1×10 7  vp/mL to about 1×10 13  vp/mL, wherein the buffer is selected from a group of buffers acceptable for human parenteral use, preferably a Tris buffer, at a pH from about 7.5 to about 8.5.  
     
     
         28 . A virus formulation of  claim 27  wherein the sugar is sucrose at a weight to volume percentage from about 2% to about 7.5% and the salt is sodium chloride from about 25 mM to about 250 mM, such that the total osmolarity of the formulation is a range from about 200 mOs/L to about 800 mOs/L.  
     
     
         29 . A virus formulation of  claim 28  wherein the divalent cation is selected from the group consisting of MgCl 2 , and CaCl 2  in an amount from about 0.1 mM to about 5 mM.  
     
     
         30 . A virus formulation of  claim 29  wherein the non-ionic detergent is selected from the group consisting of Polysorbate-80 and Polysorbate-40 at a concentration range from about 0.001% to about 2%.  
     
     
         31 . A virus formulation of  claim 30  with an adenovirus concentration in the range from about 1×10 7  vp/mL to about 1×10 13  vp/mL and a total osmolarity in a range from about 200 mOs/L to about 800 mOs/L which comprises a about 5.0 mM Tris, at pH 8.0; sucrose in a weight to volume range of about 5%; NaCl at about 75 mM, MgCl 2  at about 1 mM to 2 mM, and either Polysorbate-80 at a concentration of about 0.02% or Polysorbate-40 at a concentration of about 0.005%.  
     
     
         32 . A virus formulation of  claim 31  wherein the formulation is buffered with about 5.0 mM Tris-HCl, at pH 8.0; sucrose is present a about 5%, NaCl is present at about 75 mM, MgCl 2  at 1 mM, and Polysorbate-80 at 0.001% with the total osmolarity approximately 310 mOs/L.  
     
     
         33 . A virus formulation of  claim 25  which further comprises plasmid DNA at a concentration from about 0.01 mg/mL to about 10 mg/mL.  
     
     
         34 . A virus formulation of  claim 26  which further comprises plasmid DNA at a concentration from about 0.01 mg/mL to about 10 mg/mL.  
     
     
         35 . A method of preserving a purified virus wherein a formulation is generated which comprises the purified virus, a buffer, a sugar, a salt, a divalent cation, and a non-ionic detergent.  
     
     
         36 . The method of  claim 35  wherein the purified virus is adenovirus with a concentration in the range from about 1×10 7  vp/mL to about 1×10 13  vp/mL, wherein the buffer is selected from a group of buffers acceptable for human parenteral use, preferably a Tris buffer, at a pH from about 7.5 to about 8.5.  
     
     
         37 . The method of  claim 36  wherein the sugar is sucrose at a weight to volume percentage from about 2% to about 7.5% and the salt is sodium chloride from about 25 mM to about 250 mM, such that the total osmolarity of the formulation is a range from about 200 mOs/L to about 800 mOs/L.  
     
     
         38 . The method of  claim 37  wherein the virus formulation is selected from the group consisting of formulation number A105, A110, A111, A112, A121, A126, A127, A128, A129, A130, A131, A155, A159, A160, A165, A167, A168, A169, A170, A171, A172 and A173.  
     
     
         39 . A method of preserving a purified virus wherein a formulation is generated which comprises the purified virus, a buffer, a sugar, a salt, a divalent cation, non-ionic detergent, and an inhibitor of free radical oxidation.  
     
     
         40 . The method of  claim 39  wherein the purified virus is adenovirus with a concentration in the range from about 1×10 7  vp/mL to about 1×10 13  vp/mL, wherein the buffer is selected from a group of buffers acceptable for human parenteral use, preferably a Tris buffer, at a pH from about 7.5 to about 8.5.  
     
     
         41 . The method of  claim 40  wherein the sugar is sucrose at a weight to volume percentage from about 2% to about 7.5% and the salt is sodium chloride from about 25 mM to about 250 mM, such that the total osmolarity of the formulation is a range from about 200 mOs/L to about 800 mOs/L.  
     
     
         42 . The method of  claim 41  wherein the inhibitor of free radical oxidation is selected from the group consisting of ethanol, EDTA, an EDTA/ethanol combination, triethanolamine, and sodium citrate.  
     
     
         43 . The method of  claim 42  wherein the virus formulation is selected from the group consisting of formulation number, A113, A114, A115, A116, A117, A118, A119, A120, A112, A121, A132, A133, A134, A135, A136, A149, A151a, A151b, A152 and A153.  
     
     
         44 . The method of  claim 36  wherein the virus formulation further comprises plasmid DNA at a concentration from about 0.01 mg/mL to about 10 mg/mL.  
     
     
         45 . The method of  claim 37  wherein the virus formulation further comprises plasmid DNA at a concentration from about 0.01 mg/mL to about 10 mg/mL.