Nucleic acid delivery

Abstract

The present invention provides formulations and methods to enhance the delivery of nucleic acids to cells. Formulations comprising dextrin polymers in combination with sugars provide enhanced delivery of nucleic acids, particularly eucaryotic expression vectors, demonstrate enhanced delivery of nucleic acids to cells in vivo. The present invention also provides methods of treatment in combination with such formulations.

1 . A method to deliver nucleic acid to a cell wherein the nucleic acid is carried in a solution comprising dextrin and at least one sugar, the osmolarity of which corresponds substantially to the physiological osmolarity of the milieu surrounding the cell.  
     
     
         2 . The method according to  claim 1  wherein the molecular weight of the dextrin is in the range from about 1,000-200,000.  
     
     
         3 . The method according to  claim 2  wherein the molecular weight of the dextrin is between about 2,000-55,000.  
     
     
         4 . The method according to  claim 3  wherein the dextrin contains more than about 15% w/v of polymers of a degree of polymerisation greater than 12.  
     
     
         5 . The method according to  claim 4  wherein the dextrin contains more than about 50% w/v of polymers of a degree of polymerisation greater than 12.  
     
     
         6 . The method according to  claim 5  wherein the dextrin is present in the solution in an amount of less than about 20% w/v.  
     
     
         7 . The method according to  claim 6  wherein the dextrin is present in the solution in an amount selected from about: 1% w/v; 2% w/v; 3% w/v; 4% w/v; 5% w/v; 6% w/v; 7% w/v; 8% w/v; 9% w/v; 10% w/v; 11% w/v; 12% w/v; 13% w/v; 14% w/v; 15% w/v; 16% w/v; 17% w/v; 18% w/v; 19% w/v; 20% w/v.  
     
     
         8 . The method according to  claim 7  wherein the dextrin is present from 1-5% w/v.  
     
     
         9 . The method according to  claim 8  wherein the dextrin is preferably about 4% w/v.  
     
     
         10 . The method according to any of  claim 1  wherein the sugar is a disaccharide.  
     
     
         11 . The method according to  claim 10  wherein the amount of dissacharide is between about 1 and 10% w/v.  
     
     
         12 . The method according to  claim 11  wherein the dissacharide is an amount of between about 2 and 5% w/v.  
     
     
         13 . The method according to  claim 12  wherein the dissacharide is sucrose and the amount of sucrose is about 3% w/v.  
     
     
         14 . The method according to  claim 1  wherein the amount of dextrin is about between 2%-20% w/v and the amount of sucrose is between about 1-10% w/v.  
     
     
         15 . The method according to  claim 14  wherein the amount of dextrin is about 15% w/v and the amount of sucrose is about 3% w/v.  
     
     
         16 . The method according to  claim 15  wherein the amount of dextrin is about 4% w/v and the amount of sucrose is about 3% w/v.  
     
     
         17 . The method according to  claim 1  wherein the solution further comprises a divalent cation.  
     
     
         18 . The method according to  claim 17  wherein the divalent cation is in a concentration of at least 0.2 mM.  
     
     
         19 . The method according to  claim 19  wherein the divalent cation concentration is between 0.2-3.0 mM.  
     
     
         20 . The method according to  claim 19  wherein the divalent cation is provided by MgCl 2  and the concentration is about 2.0 mM.  
     
     
         21 . The method according to  claim 20  wherein the solution comprises about 4% w/v dextrin, about 3% w/v sucrose and about 2.0 mM MgCl 2 .  
     
     
         22 . The method according to  claim 1  wherein the nucleic acid molecule is a vector.  
     
     
         23 . The method according to  claim 22  wherein the vector is adapted for eukaryotic expression.  
     
     
         24 . The method according to  claim 22  wherein the vector is a recombinant virus derived from adenovirus, retrovirus, adeno-associated virus, herpesvirus, lentivirus, vaccinia virus, or baculovirus.  
     
     
         25 . The method of  claim 24  wherein said vector is an adenovirus.  
     
     
         26 . The method of  claim 25  wherein said adenovirus further encodes an exogenous transgene.  
     
     
         27 . The method of  claim 26  wherein said transgene is a tumor suppressor gene.  
     
     
         28 . The method of  claim 27  wherein said tumor suppressor gene is p53.  
     
     
         29 . The method of claim  claims 24  to  26  wherein said vector is replication competent.  
     
     
         30 . The method of  claim 29  wherein said vector is a conditionally replicating replication competent vector.  
     
     
         31 . A composition for delivery of a recombinant viral vector to a cell comprising a recombinant viral vector in a solution comprising dextrin and at least one sugar, the osmolarity of which corresponds substantially to the physiological osmolarity of the milieu surrounding the cell.  
     
     
         32 . The composition of  claim 31  wherein the recombinant viral vector is derived from adenovirus, retrovirus, adeno-associated virus, herpesvirus, lentivirus, vaccinia virus, or baculovirus.  
     
     
         33 . The composition of  claim 32  wherein the viral vector is derived from adenovirus.  
     
     
         34 . The composition of  claim 33  wherein the viral vector is a selectively-replicating replication competent adenoviral vector.  
     
     
         35 . A pharmaceutical formulation comprising a recombinant adenoviral vector, a dextrin, a sugar and a divalent cation.  
     
     
         36 . A method to deliver nucleic acid to a cell comprising: providing a solution of dextrin, a sugar, a divalent cation and nucleic acid and contacting the solution with a cell to be transfected or transformed.