Interfering Rna Duplex Having Blunt-Ends And 3'-Modifications

Abstract

The present invention relates to double-stranded RNA compounds with at least one blunt end comprising at least one 3′-end of Formula (I): wherein X is O or S R 1 and R 2 are independently OH, NH 2 , SH, alkyl, aryl, alkyl-aryl, aryl-alkyl, where alkyl, aryl, alkyl-aryl, aryl-alkyl can be substituted by additional heteroatoms and functional groups, preferably a heteroatom selected from the group of N, O, or S or a functional group selected from the group OH, NH 2 , SH, carboxylic acid or ester; or R 1 and R 2 may be of formula Y-Z where Y is O, N, S and Z is H, alkyl, aryl, alkyl-aryl, aryl-alkyl, where alkyl, aryl, alkyl-aryl, aryl-alkyl can be substituted by additional heteroatoms, preferably a heteroatom selected from the group of N, O, or S; and wherein said double-stranded RNA mediates RNA interference. Preferred 3′end modifications are: phosphate, phosphorothioate, abasic ribonucleoside, hydroxyphopyl phosphodiester.

1 . Double-stranded RNA with at least one blunt end comprising at least one 3′-end of formula:  
       
         
           
           
               
               
           
         
       
       wherein 
 X is O or S  
 R 1  and R 2  are independently OH, NH 2 , SH, alkyl, aryl, alkyl-aryl, aryl-alkyl, where alkyl, aryl, alkyl-aryl, aryl-alkyl can be substituted by additional heteroatoms and functional groups, preferably a heteroatom selected from the group of N, O, or S or a functional group selected from the group OH, NH 2 , SH, carboxylic acid or ester;  
 or R 1  and R 2  may be of formula Y-Z where Y is O, N, S and Z is H, alkyl, aryl, alkyl-aryl, aryl-alkyl, where alkyl, aryl, alkyl-aryl, aryl-alkyl can be substituted by additional heteroatoms, preferably a heteroatom selected from the group of N, O, or S; and wherein said double-stranded RNA mediates RNA interference.  
 
     
     
         2 . Double-stranded RNA according to  claim 1 , wherein R 1  is OH.  
     
     
         3 . Double-stranded RNA according to  claim 1  comprising a region of 15 to 30 nucleotides complementary to a target gene.  
     
     
         4 . Double-stranded RNA according to  claim 1  wherein R 1  and R 2  are independently OH, NH 2 , SH, lower alkyl, lower aryl, lower alkyl-aryl, lower aryl-alkyl, where lower alkyl, lower aryl, lower alkyl-aryl, lower aryl-alkyl can be substituted by additional heteroatoms and functional groups, preferably a heteroatom selected from the group of N, O or S or a functional group selected from the group OH, NH 2 , SH, carboxylic acid or ester; or, R 1  and R 2  may be of formula Y-Z where Y is O, N, S, B and Z is alkyl, aryl, alkyl-aryl, aryl-alkyl, where alkyl, aryl, alkyl-aryl, aryl-alkyl can be substituted by additional heteroatoms, preferably a heteroatom selected from the group of N, O, or S, provided that not both R 1  and R 2  are OH.  
     
     
         5 . Double-stranded RNA according to  claim 1  wherein R 1  is OH and R 2  comprises from 1 to 24 C-atoms.  
     
     
         6 . Double-stranded RNA according to any of  claim 1  wherein said RNA comprises two blunt ends.  
     
     
         7 . Double-stranded RNA according to  claim 1  wherein Z is one or more abasic nucleosides.  
     
     
         8 . A method for the inhibition of a target gene comprising introducing into a cell a dsRNA according to any of claims  1 .  
     
     
         9 . The method of  claim 7  wherein the target gene is a pathogen-associated gene, a viral gene or an oncogene.  
     
     
         10 . A kit comprising reagents for inhibiting expression of a target gene in a cell, wherein said kit comprises dsRNA according to any of claims  1  into a cell and a means for introduction of a dsRNA into a cell in an amount sufficient to inhibit expression of the target gene.  
     
     
         11 . A pharmaceutical composition comprising an effective amount of at least one dsRNA according to any of claims  1  for the inhibition of a target gene.  
     
     
         12 . A method for identifying and/or characterizing pharmacological agents acting on at least one target protein comprising: contacting a eukaryotic cell capable of expressing at least one endogenous gene coding for the protein(s) of interest with 
 (a) at least one dsRNA molecule according to the present invention, which is capable of inhibiting the expression of the gene(s) encoding the protein(s) of interest and    (b) a test substance or a collection of test substances wherein pharmacological properties of said test substance or said collection are to be identified and/or characterized.    
     
     
         13 . Use of a double stranded RNA according to claims  1  for the inhibition of a target gene.