Modified iRNA agents
The invention relates to iRNA agents, which preferably include a monomer in which the ribose moiety has been replaced by a moiety other than ribose.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.
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 or NR 7 ;
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 R 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 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.