Compounds and methods to enhance rAAV transduction

Abstract

Agents and methods to alter rAAV transduction are provided.

1 - 12 . (canceled) 
     
     
         13 . A method to enhance adeno-associated virus (AAV) transduction of a mammalian lung or liver cell, comprising: contacting the mammalian lung or liver cell with an amount of AAV and an amount of an agent effective to enhance AAV transduction. 
     
     
         14 . (canceled) 
     
     
         15 . A method to enhance the expression of a transgene in a mammalian lung or liver cell, comprising: contacting the mammalian lung or liver cell with an amount of an agent and an amount of recombinant AAV (rAAV) comprising the transgene so as to enhance expression of the transgene. 
     
     
         16 . (canceled) 
     
     
         17 . A method comprising: administering to a mammal subjected to gene therapy with rAAV comprising a transgene, an amount of an agent effective to enhance expression of the transgene in the cells of the mammal, wherein the agent enhances AAV transduction after viral binding to the membrane of a mammalian cell and before second strand synthesis which yields an expressible form of the viral genome. 
     
     
         18 . The method of  claim 13 ,  15 , or  17  wherein endosomal processing of the virus is altered. 
     
     
         19 . The method of  claim 13  wherein the virus is rAAV. 
     
     
         20 . The method of  claim 19  wherein the rAAV encodes a therapeutic peptide or polypeptide. 
     
     
         21 . The method of  claim 15  wherein the rAAV encodes a therapeutic peptide or polypeptide. 
     
     
         22 . The method of  claim 17  wherein the rAAV encodes a therapeutic peptide or polypeptide. 
     
     
         23 . The method of  claim 13 , or  15 , wherein the cell is contacted with the agent before the cell is contacted with the virus. 
     
     
         24 . The method of  claim 13 , or  15 , wherein the cell is contacted with the virus before the cell is contacted with the agent. 
     
     
         25 - 26 . (canceled) 
     
     
         27 . The method of  claim 17  wherein expression is enhanced in lung cells. 
     
     
         28 . The method of  claim 17  wherein expression is enhanced in liver cells. 
     
     
         29 - 84 . (canceled) 
     
     
         85 . A method to enhance AAV transduction of a mammalian cell, comprising:
 contacting the mammalian cell with an amount of AAV and an amount of an agent effective to enhance AAV transduction, wherein the agent enhances AAV transduction after viral binding to the membrane of a mammalian cell and before second strand synthesis which yields an expressible form of the viral genome.   
     
     
         86 . A method to enhance AAV transduction in a mammal, comprising: administering to a mammal in need of therapy an effective amount of rAAV encoding a therapeutic gene product and an agent, wherein the agent enhances AAV transduction after viral binding to the membrane of a mammalian cell and before second strand synthesis which yields an expressible form of the viral genome. 
     
     
         87 . The method of  claim 85  or  86  wherein the agent enhances endosomal processing. 
     
     
         88 . The method of  claim 85  or  86  wherein the agent is an endosomal protease inhibitor. 
     
     
         89 . The method of  claim 85  or  86  wherein the agent is a cysteine protease inhibitor. 
     
     
         90 . The method of  claim 85  or  86  wherein the agent is a peptide or analog thereof. 
     
     
         91 . The method of  claim 85  or  86  wherein the agent is a compound of formula (I): R 1 -A-(B) n —C wherein R 1  is an N-terminal amino acid blocking group; each A and B is independently an amino acid; C is an amino acid wherein the terminal carboxy group has been replaced by a formyl (CHO) group; and n is 0, 1, 2, or 3; or a pharmaceutically acceptable salt thereof. 
     
     
         92 . The method of  claim 85  or  86  wherein the agent is a compound of formula (II): 
       
         
           
           
               
               
           
         
       
       wherein
 R 2  is an N-terminal amino acid blocking group; 
 R 3 , R 4 , and R 5  are each independently hydrogen, (C 1 -C 10 )alkyl, aryl or aryl(C 1 -C 10 )alkyl; and 
 R 6 , R 7 , and R 8  are each independently hydrogen, (C 1 -C 10 )alkyl, aryl or aryl(C 1 -C 10 )alkyl; 
 
       or a pharmaceutically acceptable salt thereof. 
     
     
         93 . The method of  claim 13 ,  15 ,  17 ,  85  or  86  wherein the agent enhances AAV transport to the nucleus. 
     
     
         94 . The method of  claim 17  or  86  wherein the rAAV and the agent are concurrently administered. 
     
     
         95 . A method to enhance AAV transduction of a mammalian lung or liver cell, comprising: contacting the mammalian lung or liver cell with AAV and an effective amount of an agent selected from the group consisting of a compound of formula (I): R 1 -A-(B) n —C wherein R 1  is an N-terminal amino acid blocking group; each A and B is independently an amino acid; C is an amino acid wherein the terminal carboxy group has been replaced by a formyl (CHO) group; and n is 0, 1, 2, or 3; or a pharmaceutically acceptable salt thereof;
 a compound of formula (II):   
       
         
           
           
               
               
           
         
       
       wherein
 R 2  is an N-terminal amino acid blocking group; 
 R 3 , R 4 , and R 5  are each independently hydrogen, (C 1 -C 10 )alkyl, aryl or aryl(C 1 -C 10 )alkyl; and 
 R 6 , R 7 , and R 8  are each independently hydrogen, (C 1 -C 10 )alkyl, aryl or aryl(C 1 -C 10 )alkyl; 
 
       or a pharmaceutically acceptable salt thereof;
 a compound of formula (III): 
 
       
         
           
           
               
               
           
         
       
       wherein
 R 1  is H, halogen, (C 1 -C 10 )alkyl, (C 1 -C 10 )alkenyl, (C 1 -C 10 )alkynyl, (C 1 -C 10 )alkoxy, (C 1 -C 10 )alkanoyl, (═O), (═S), OH, SR, CN, NO 2 , or trifluoromethyl, wherein any alkyl, alkenyl, alkynyl, alkoxy or alkanoyl may optionally be substituted with one or more halogen, OH, SH, CN, NO 2 , trifluoromethyl, NRR or SR, wherein each R is independently H or (C 1 -C 10 )alkyl; 
 R 2  is (═O) or (═S); 
 R 3  is H, (C 1 -C 10 )alkyl, (C 1 -C 10 )alkenyl, (C 1 -C 10 )alkynyl, (C 1 -C 10 )alkoxy or (C 3 -C 8 )cycloalkyl, wherein any alkyl, alkenyl, alkynyl, alkoxy or cycloalkyl may optionally be substituted with one or more halogen, OH, CN, NO 2 , trifluoromethyl, SR, or NRR, wherein each R is independently H or (C 1 -C 10 )alkyl; 
 R 4  is H, (C 1 -C 10 )alkyl, (C 1 -C 10 )alkenyl, (C 1 -C 10 )alkynyl, (C 1 -C 10 )alkoxy or (C 3 -C 8 )cycloalkyl, wherein any alkyl, alkenyl, alkynyl, alkoxy or cycloalkyl may optionally be substituted with one or more halogen, OH, CN, NO 2 , trifluoromethyl, SR, or NRR, wherein each R is independently H or (C 1 -C 10 )alkyl; 
 R 5  is H, halogen, (C 1 -C 10 )alkyl, (C 1 -C 10 )alkenyl, (C 1 -C 10 )alkynyl, (C 1 -C 10 )alkoxy, (C 1 -C 10 )alkanoyl, (═O), (═S), OH, SR, CN, NO 2  or trifluoromethyl, wherein any alkyl, alkenyl, alkynyl, alkoxy or alkanoyl may optionally be substituted with one or more halogen, OH, SH, CN, NO 2 , trifluoromethyl, NRR or SR, wherein each R is independently H or (C 1 -C 10 )alkyl; and 
 X is O, S or NR wherein R is H or (C 1 -C 10 )alkyl, or a pharmaceutically acceptable salt thereof; and 
 a compound of formula (IV):
   R-A-A 1 -R 1    
 
 
       wherein R is hydrogen, an amino acid, or a peptide, wherein the N-terminus amino acid can optionally be protected at the amino group with acetyl, acyl, trifluoroacetyl, or benzyloxycarbonyl; A is an amino acid or a direct bond; A 1  is an amino acid; and R 1  is hydroxy or an amino acid, wherein the C-terminus amino acid can optionally be protected at the carboxy group with (C 1 -C 6 )alkyl, phenyl, benzyl ester or amide; 
       or a pharmaceutically acceptable salt thereof.