Use of a nuclease inhibitor or interleukin-10 (IL-10) for the preparation of a therapeutic composition for improving transfection of a polynucleotide into a cell and compositions useful in gene therapy

Abstract

Described is the use of a nuclease inhibitor or of interleukin-10 (IL-10) for the preparation of a therapeutic composition for improving transfection of a polynucleotide into a cell, and to compositions comprising a mixture of polynucleotide and nuclease inhibitor and/or interleukin-10.

1 . Use of a nuclease inhibitor for the preparation of a therapeutic composition for the introduction of a polynucleotide into a cell.  
     
     
         2 . The use of  claim 1 , wherein said nuclease inhibitor is a deoxyribonuclease (DNAse) inhibitor, preferably a DNAse I inhibitor.  
     
     
         3 . The use of  claim 2 , wherein said nuclease inhibitor is G-actin or a fragment thereof capable of inhibiting DNAse I activity.  
     
     
         4 . The use of  claim 3 , wherein said G-actin is of porcine, rabbit, bovine, simian, murine or human origin.  
     
     
         5 . The use of any one of  claims 1  to  4 , wherein said therapeutic composition is for administration into a vertebrate target tissue.  
     
     
         6 . The use of  claim 5 , wherein said administration is made by intradermal, subdermal, intravenous, intramuscular, intranasal, intracerebral, intratracheal, intraarterial, intraperitoneal, intravesical, intrapleural, intracoronary or intratumoral injection.  
     
     
         7 . The use of  claim 5 , wherein said administration is made into the lung by inhalation or aerosol administration.  
     
     
         8 . The use of  claim 5 , wherein said target tissue is muscle.  
     
     
         9 . The use of any one of  claims 5  to  8 , wherein the administration of the nuclease inhibitor is performed independently from a second administration consisting in administration of a composition containing at least one polynucleotide into the same target tissue.  
     
     
         10 . The use of  claim 9 , wherein the administration of the nuclease inhibitor is performed prior to said second administration.  
     
     
         11 . The use of any one of  claims 1  to  8 , wherein said therapeutic composition further comprises at least one polynucleotide.  
     
     
         12 . The use of any one of  claims 1  to  11 , wherein said polynucleotide contains a gene and is capable of functionally expressing said gene in said cell.  
     
     
         13 . A composition for introducing a polynucleotide into a cell, said composition comprising at least one polynucleotide and at least one nuclease inhibitor.  
     
     
         14 . The composition of  claim 13 , wherein said nuclease inhibitor is a DNAse inhibitor, preferably a DNAse I inhibitor.  
     
     
         15 . The composition of  claim 14 , wherein said nuclease inhibitor is G-actin or a fragment thereof with the capability to inhibit DNAse I activity.  
     
     
         16 . The composition of  claim 15 , wherein said G-actin or fragment thereof is porcine, rabbit, bovine or human origin.  
     
     
         17 . The composition of  claim 15  or 16, wherein said composition contains between 4×10 −5  and 4 μg, preferably between 4×10 −4  and 2 μg, and more preferably comprises between 4×10 −3  and 4×10 −1  μg of nuclease inhibitor per μg of DNA.  
     
     
         18 . The composition of any one of  claims 13  to  17 , wherein the polynucleotide concentration ranges from about 0.1 μg/ml to about 20 mg/ml.  
     
     
         19 . The composition of any one of  claims 13  to  18 , wherein said polynucleotide contains a gene and is capable of functionally expressing said gene in said cell.  
     
     
         20 . The composition of  claim 18 , wherein said gene encodes all or part of dystrophin or cystic fibrosis transmembrane conductance regulator (CFTR) polypeptides.  
     
     
         21 . The composition of any one of  claims 13  to  20 , wherein said cell is a vertebrate cell.  
     
     
         22 . The composition of any one of  claims 13  to  21 , wherein said polynucleotide is naked.  
     
     
         23 . The composition of any one of  claims 13  to  21 , wherein said polynucleotide is associated with viral polypeptides.  
     
     
         24 . The composition of any one of  claims 13  to  21 , wherein said polynucleotide is complexed with cationic components, more preferably with cationic lipids.  
     
     
         25 . The composition of any of  claims 13  to  24 , wherein said composition further comprises at least one component selected from the group consisting of chloroquine, protic compounds such as propylene glycol, polyethylene glycol, glycerol, ethanol, 1-methyl L-2-pyrrolidone or derivatives, aprotic compounds such as dimethylsulfoxide (DMSO), diethylsulfoxide, di-n-propylsulfoxide, dimethylsulfone, sulfolane, dimethylformamide, dimethylacetamide, tetramethylurea, acetonitrile or derivatives, cytokines, preferably interleukin 10 (IL-10).  
     
     
         26 . The composition of  claim 25 , wherein said composition comprises 5-15% of DMSO and/or from about 0.001 to about 1 μg preferably from about 0.01 to about 0.1 μg of IL-10.  
     
     
         27 . The composition of any of  claims 13  to  26  for use in a method for the therapeutic treatment of the human or animal body.  
     
     
         28 . The composition of  claim 27 , wherein said composition further comprises a pharmaceutically acceptable injectable carrier.  
     
     
         29 . A process for introducing a polynucleotide into cells wherein said process comprises contacting said cells with at least one composition of any one of  claims 13  to  28 .  
     
     
         30 . A process for introducing a polynucleotide into cells wherein said process comprises contacting the cells simultaneously or subsequently with a nuclease inhibitor and the polynucleotide.  
     
     
         31 . The process of  claim 30 , wherein the cells are first contacted with the nuclease inhibitor and subsequently with the polynucleotide.  
     
     
         32 . Use of interleukin-10 (IL-10) for the preparation of a therapeutic composition for the introduction of a polynucleotide into a cell.  
     
     
         33 . The use of  claim 32 , wherein said interleukin-10 is of human, simian, rabbit, bovine, porcine or murine origin.  
     
     
         34 . The use of claims  32  and  33 , wherein said therapeutic composition is for administration into a vertebrate target tissue.  
     
     
         35 . The use of  claim 34 , wherein said administration is made by intradermal, subdermal, intravenous, intramuscular, intranasal, intracerebral, intratracheal, intraarterial, intraperitoneal, intravesical, intrapleural, intracoronary or intratumoral injection.  
     
     
         36 . The use of  claim 34 , wherein said administration is made into the lung by inhalation or aerosol administration.  
     
     
         37 . The use of  claim 34 , wherein said target tissue is muscle.  
     
     
         38 . The use of any one of  claims 34  to  37 , wherein the administration of the interleukin-10 is performed independently from a second administration consisting in administration of a composition containing at least one polynucleotide into the same target tissue.  
     
     
         39 . The use of  claim 38 , wherein the administration of the interleukin-10 is performed prior to said second administration.  
     
     
         40 . The use of any one of  claims 32  to  37 , wherein said therapeutic composition further comprises at least one polynucleotide.  
     
     
         41 . The use of any one of  claims 32  to  40 , wherein said polynucleotide contains a gene and is capable of functionally expressing said gene in said cell.  
     
     
         42 . A composition for introducing a polynucleotide into a cell, said composition comprising an IL-10 and at least one polynucleotide.  
     
     
         43 . The composition of  claim 42 , wherein said interleukin-10 is of human, simian, rabbit, bovine, porcine or murine origin.  
     
     
         44 . The composition of  claim 42  or  43 , wherein said composition contains between from about 0.001 to about 1 μg, preferably from about 0.01 to about 0.1 μg of IL-10.  
     
     
         45 . The composition of any one of  claims 42  to  44 , wherein the polynucleotide concentration ranges from about 0.1 μg/ml to about 20 mg/ml.  
     
     
         46 . The composition of any one of  claims 42  to  45 , wherein said polynucleotide contains a gene and is capable of functionally expressing said gene in said cell.  
     
     
         47 . The composition of  claim 46 , wherein said gene encodes all or part of dystrophin or cystic fibrosis transmembrane conductance regulator (CFTR) polypeptides.  
     
     
         48 . The composition of any one of  claims 42  to  47 , wherein said cell is a vertebrate cell.  
     
     
         49 . The composition of any one of  claims 42  to  48 , wherein said polynucleotide is naked.  
     
     
         50 . The composition of any one of  claims 42  to  49 , wherein said polynucleotide is associated with viral polypeptides.  
     
     
         51 . The composition of any one of  claims 42  to  48 , wherein said polynucleotide is complexed with cationic components, more preferably with cationic lipids.  
     
     
         52 . The composition of any of  claims 42  to  51 , wherein said composition further comprises at least one component selected from the group consisting of chloroquine, protic compounds such as propylene glycol, polyethylene glycol, glycerol, ethanol, 1-methyl L-2-pyrrolidone or derivatives, aprotic compounds such as dimethylsulfoxide (DMSO), diethylsulfoxide, di-n-propylsulfoxide, dimethylsulfone, sulfolane, dimethylformamide, dimethylacetamide, tetramethylurea, acetonitrile or derivatives, cytokines different from interleukin 10 (IL-10).  
     
     
         53 . The composition of  claim 52 , wherein said composition further comprises 5-15% of DMSO.  
     
     
         54 . The composition of any of  claims 42  to  53  for use in a method for the therapeutic treatment of the human or animal body.  
     
     
         55 . The composition of  claim 54 , wherein said composition further comprises a pharmaceutically acceptable injectable carrier.  
     
     
         56 . A process for introducing a polynucleotide into cells wherein said process comprises contacting said cells with at least one composition of any one of  claims 52  to  55 .  
     
     
         57 . A process for introducing a polynucleotide into cells wherein said process comprises contacting the cells with said polynucleotide prior to, concurrent with or subsequent to contacting them with interleukin-10.  
     
     
         58 . The process of  claim 57 , wherein the cells are first contacted with the interleukin-10 and subsequently with the polynucleotide.