Chemically modified oligonucleotides
View Patent ↗This invention relates composition and methods for making and using chemically modified oligonucleotides agents for inhibiting gene expression.
1. An isolated oligonucleotide agent, comprising a nucleotide sequence consisting of from 12 to 23 nucleotides in length sufficiently complementary to a microRNA target sequence of about 12 to 23 nucleotides, wherein the nucleotide sequence of the oligonucleotide agent differs by no more than 1 or 2 nucleotides from full complementary to the microRNA target sequence and wherein said oligonucleotide agent has the structure (I)
(5′) QxQz 1 (Qy) n Qz 2 Qz 3 Qz 4 Q-L (3′) (I)
wherein
Q is a 2′-O-methyl modified nucleoside;
x, z 1 , z 2 , z 3 , and z 4 are all
One of A and B is S while the other is O;
n=6-17;
L is
wherein:
X is N(CO)R 7 , or NR 7 ;
each of R 1 , R 3 and R 9 , is, independently, H, OH, or —CH 2 OR b provided that at least one of R 1 , R 3 , or R 9 is OH and at least one of R 1 , R 3 or R 9 is —CH 2 OR b ;
R 7 is C 1 -C 20 alkyl substituted with NR c R d or NHC(O)R d ;
R c is H or C 1 -C 6 alkyl;
R d is a carbohydrate radical; or a steroid radical, which is optionally tethered to at least one carbohydrate radical; and
R b is
one of E and F is S while the other is O.
2. The oligonucleotide agent of claim 1 , wherein R d is cholesterol.
3. The compound of claim 1 , wherein R 1 is —CH 2 OR b .
4. The compound of claim 3 , wherein R 9 is OH.
5. The compound of claim 4 , wherein R 1 and R 9 are trans.
6. The compound of claim 1 , wherein R 3 is OH.
7. The compound of claim 6 , wherein R 1 and R 3 are trans.
8. The compound of claim 1 , wherein R 3 is —CH 2 OR b .
9. The compound of claim 8 , wherein R 1 is OH.
10. The compound of claim 9 , wherein R 1 and R 3 are trans.
11. The compound of claim 8 , wherein R 9 is OH.
12. The compound of claim 11 , wherein R 3 and R 9 are trans.
13. The compound of claim 1 , wherein R 9 is —CH 2 OR b .
14. The compound of claim 13 , wherein R 1 is OH.
15. The compound of claim 14 , wherein R 1 and R 9 are trans.
16. The compound of claim 1 , wherein X is NC(O)R 7 .
17. The compound of claim 16 , wherein R 7 is —CH 2 (CH 2 ) 3 CH 2 NHC(O)R d .
18. The oligonucleotide agent of claim 1 , wherein R 1 is CH 2 OR b ; R 9 is OH; R 1 and R 9 are trans; X is NC(O)R 7 ; R 7 is CH 2 (CH 2 ) 3 CH 2 NHC(O)R d and R d is a steroid radical.
19. The oligonucleotide agent of claim 18 , wherein the nucleotide sequence of the oligonucleotide agent is SEQ ID NO:96.
20. The oligonucleotide agent of claim 18 , wherein said oligonucleotide agent consists of a sequence that differs at no more than 1 or 2 nucleotides from a sequence of 12 or more contiguous nucleotides of SEQ ID NO:96.
21. The oligonucleotide agent of claim 18 , wherein the nucleotide sequence of the oligonucleotide agent is SEQ ID NO:101.
22. The oligonucleotide agent of claim 18 , wherein the nucleotide sequence of the oligonucleotide agent is SEQ ID NO:102.
23. The oligonucleotide agent of claim 18 , wherein the nucleotide sequence of the oligonucleotide agent is SEQ ID NO:103.