MODIFIED iRNA AGENTS

Abstract

The invention relates to iRNA agents, which preferably include a monomer in which the ribose moiety has been replaced by a moiety other than ribose. The inclusion of such a monomer can allow for modulation of a property of the iRNA agent into which it is incorporated, e.g., by using the non-ribose moiety as a point to which a ligand or other entity, e.g., a lipophilic moiety. e.g., cholesterol, is is directly, or indirectly, tethered. The invention also relates to methods of making and using such modified iRNA agents.

What is claimed is: 
     
         1 . An RNA agent comprising a sense strand and an antisense strand, wherein one or more ribose replacement modification subunit (RRMS) comprising a ligand is incorporated into at least one of said strands, and wherein the RRMS is a cyclic carrier selected from the group consisting of hydroxyproline, piperidine, morpholine, piperazine, and decalin, and wherein the ligand is connected to the RRMS via a tethering moiety containing a C 1 -C 20  alkyl group and having at least one nitrogen atom. 
     
     
         2 . The RNA agent of  claim 1 , wherein the tethering moiety is a C 1 -C 20  alkyl group substituted with a —NHC(O)— group. 
     
     
         3 . The RNA agent of  claim 2 , wherein the tethering moiety is a C 1 -C 10  alkyl group substituted with a —NHC(O)— group. 
     
     
         4 . The RNA agent of  claim 1 , wherein the ligand comprises one or more carbohydrate moieties. 
     
     
         5 . The RNA agent of  claim 4 , wherein the carbohydrate moieties are monosaccharides, disaccharides, trisaccharides, tetrasaccharides, polysaccharides, or combinations thereof. 
     
     
         6 . The RNA agent of  claim 1 , wherein the ligand is selected from the group consisting of a mucin carbohydrate, multivalent lactose, multivalent galactose, N-acetyl-galactosamine, multivalent N-acetyl-galactosamine, N-acetyl-glucosamine, multivalent N-acetyl-glucosamine, multivalent mannose, multivalent fucose, glycosylated polyaminoacids, lactose, galactose, mannose, and combinations thereof. 
     
     
         7 . The RNA agent of  claim 1 , wherein the RRMS subunit is placed within 1, 2, or 3 positions of the 3′ or 5′ end of at least one of said strands. 
     
     
         8 . The RNA agent of  claim 1 , wherein at least two RRMS subunits are incorporated into at least one of said strands. 
     
     
         9 . The RNA agent of  claim 8 , wherein at least three RRMS subunits are incorporated into at least one of said strands. 
     
     
         10 . An RNA agent comprising a sense strand and an antisense strand, wherein at least one subunit having a formula (I) is incorporated into at least one of said strands: 
       
         
           
           
               
               
           
         
         wherein:
 X is N(CO)R 7 , NR 7 , or CH 2 , 
 Y is NR 8 , O, S, CR 9 R 10 , or absent; 
 Z is CR 11 R 12  or absent; 
 each of R 1 , R 2 , R 3 , R 4 , R 9 , and R 10  is, independently, H, OR a , OR b , (CH 2 ) n OR a , or (CH 2 ) n OR b , provided that at least one of R 1 , R 2 , R 3 , R 4 , R 9 , and R 10  is OR a  or OR b  and that at least one of R 1 , R 2 , R 3 , R 4 , R 9 , and R 10  is (CH 2 ) n OR a  or (CH 2 ) n OR b ; 
 each of R 5 , R 6 , R 11 , and R 12  is, independently, H, C 1 -C 6  alkyl optionally substituted with 1-3 R 13 , or C(O)NHR 7 ; or R 5  and R 11  together are C 3 -C 8  cycloalkyl optionally substituted with R 14 ; 
 R 7  is a ligand tethered through a tethering moiety containing a C 1 -C 20  alkyl group and having at least one nitrogen atom; 
 R 8  is C 1 -C 6  alkyl; 
 R 13  is hydroxy, C 1 -C 4  alkoxy, or halo; 
 R 14  is NR c R 7 ; 
 R a  is H or 
 
       
       
         
           
           
               
               
           
         
         
           R b  is H or 
         
       
       
         
           
           
               
               
           
         
         wherein the Strand in each occurrence is independently the sense strand or antisense strand of the iRNA agent;
 each of A and C is, independently, O or S; 
 B is OH, O − , or 
 
       
       
         
           
           
               
               
           
         
         
           R c  is H or C 1 -C 6  alkyl; and 
           n is 1-4. 
         
       
     
     
         11 . The RNA agent of  claim 10 , wherein X is N(CO)R 7  or NR 7 , Y is CR 9 R 10 , and Z is absent. 
     
     
         12 . The RNA agent of  claim 11 , wherein R 2 , R 3 , R 4 , R 5 , R 6 , and R 10  are H; R 1  is CH 2 OR a ; and R 9  is OR b . 
     
     
         13 . The RNA agent of  claim 10 , wherein the tethering moiety in R 7  is a C 1 -C 20  alkyl group substituted with a —NHC(O)— group. 
     
     
         14 . The RNA agent of  claim 13 , wherein the tethering moiety in R 7  is a C 1 -C 10  alkyl group substituted with a —NHC(O)— group. 
     
     
         15 . The RNA agent of  claim 10 , wherein the ligand comprises one or more carbohydrate moieties. 
     
     
         16 . The RNA agent of  claim 15 , wherein the carbohydrate moieties are monosaccharides, disaccharides, trisaccharides, tetrasaccharides, polysaccharides, or combinations thereof. 
     
     
         17 . The RNA agent of  claim 10 , wherein the ligand is selected from the group consisting of a mucin carbohydrate, multivalent lactose, multivalent galactose, N-acetyl-galactosamine, multivalent N-acetyl-galactosamine, N-acetyl-glucosamine, multivalent N-acetyl-glucosamine, multivalent mannose, multivalent fucose, glycosylated polyaminoacids, lactose, galactose, mannose, and combinations thereof. 
     
     
         18 . The RNA agent of  claim 10 , wherein the subunit having formula (I) is placed within 1, 2, or 3 positions of the 3′ or 5′ end of at least one of said strands. 
     
     
         19 . The RNA agent of  claim 10 , wherein at least two subunits having formula (I) are incorporated into at least one of said strands. 
     
     
         20 . The RNA agent of  claim 19 , wherein at least three subunits having formula (I) are incorporated into at least one of said strands.