Modified iRNA agents
View Patent ↗The invention relates to iRNA agents, which preferably include a monomer in which the ribose moiety has been replaced by a moiety other than ribose.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 directly, or indirectly, tethered. The invention also relates to methods of making and using such modified iRNA agents.
1. An iRNA agent comprising a first strand and a second strand, wherein at least two subunits having a formula (I) are 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 R d ; or C 1 -C 20 alkyl substituted with NR c R d or NHC(O)R d ;
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 first strand or second strand of the iRNA agent;
each of A and C is, independently, O or S; and
B is OH, O − , or
R c is H or C 1 -C 6 alkyl;
R d is H or a ligand; and
n is 1-4.
2. The iRNA agent of claim 1 , wherein three subunits having a formula (I) are incorporated into at least one of said strands.
3. The iRNA agent of claim 2 , wherein the three subunits having a formula (I) are placed at positions 1, 2 and 3 of the 3′ end of a sense strand.
4. The iRNA agent of claim 1 , wherein X is N(CO)R 7 or NR 7 , Y is CR 9 R 10 , and Z is absent.
5. The iRNA agent of claim 4 , wherein one of R 1 , R 2 , R 3 , R 4 , R 9 , and R 10 is OR a or OR b ; and 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 .
6. The iRNA agent of claim 4 , wherein R 2 , R 3 , R 4 , R 5 , R 6 , and R 10 are H.
7. The iRNA agent of claim 4 , wherein R 9 is OR a or OR b , and R 1 is (CH 2 ) n OR a or (CH 2 ) n OR b .
8. The iRNA agent of claim 1 , R 1 can be CH 2 OR a and R 9 can be OR b .
9. The iRNA agent of claim 8 , wherein R 1 and R 9 are cis.
10. The iRNA agent of claim 8 , wherein R 1 and R 9 are trans.
11. The iRNA agent of claim 1 , wherein R d is selected from the group consisting of a folic acid radical, a cholesterol radical, a carbohydrate radical, a vitamin A radical, a vitamin E radical, and a vitamin K radical.
12. The iRNA agent of claim 1 , wherein R d is a cholesterol radical.
13. The iRNA agent of claim 1 , wherein R d is a carbohydrate radical.
14. The iRNA agent of claim 1 , wherein R d is a steroid radical, which is optionally tethered to at least one carbohydrate radical.
15. The iRNA agent of claim 1 , wherein at least one of said strands leads to the cleavage of a complementary target sequence via a RISC-dependent mechanism.
16. The iRNA agent of claim 1 , wherein A is O in each occurrence.
17. The iRNA agent of claim 1 , wherein A is S in at least one occurrence.
18. The iRNA agent of claim 1 , wherein formula (I) is incorporated into a sense strand.
19. The iRNA agent of claim 1 , wherein formula (I) is incorporated into positions 1, 2 and 3 of the 3′ end of a sense strand.
20. The iRNA agent of claim 1 , wherein the iRNA agent is 21 nucleotides in length and has a duplex region of about 19 pairs.
21. The iRNA agent of claim 1 , wherein the iRNA agent includes a duplex region between 17 and 23 pairs in length.
22. The iRNA agent of claim 1 , wherein R d is a ligand selected from the group consisting of a lipophile, a lipid, a steroid, a terpene, a vitamin, a carbohydrate, a protein, a protein binding agent, an integrin targeting molecule, polycationic, a peptide or peptide mimic, a polyamine, a dye, an intercalating agent, a cross-linker, a porphyrin, a polycyclic aromatic hydrocarbon, an artificial endonuclease, a lipophilic molecule, a glycerol or ester or ether thereof, a geranyloxyhexyl group, a hexadecylglycerol, a borneol, a menthol, a 1,3-propanediol, a heptadecyl group, a palmitic acid, a myristic acid, an O3-(oleoyl)lithocholic acid, an O3-(oleoyl)cholenic acid, a dimethoxytrityl, a phenoxazine, a peptide conjugate, an alkylating agent, a phosphate, an amino, a mercapto, a PEG, a MPEG, a [MPEG] 2 , an alkyl or substituted alkyl, a radiolabeled marker, an enzyme, a hapten, a transport/absorption facilitator, a synthetic ribonuclease, a dinitrophenyl, a HRP, an AP, and a derivative and a combination thereof.
23. A kit comprising an iRNA agent of claim 1 , a sterile container in which the iRNA agent is enclosed, and instructions for use.