Nucleic acid chemical modifications
View Patent ↗The present invention provides nucleosides of formula (1) and oligonucleotides comprising at least one nucleoside of formula (2): Another aspect of the invention relates to a method of inhibiting the expression of a gene in cell, the method comprising (a) contacting an oligonucleotide of the invention with the cell; and (b) maintaining the cell from step (a) for a time sufficient to obtain degradation of the mRNA of the target gene.
1. A nucleoside of formula (1):
wherein:
one of D 1 and D 2 is H, or a hydroxyl protecting group, and the other of D 1 and D 2 is H, a hydroxyl protecting group, or a reactive phosphorus group, provided that when one of D 1 and D 2 is a reactive phosphorus group, the other of D 1 and D 2 is a hydroxyl protecting group;
m is an integer from 1 to 15;
B is selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, a natural nucleobase, and a universal nucleobase;
Q is selected from the group consisting of O, S, NR 10 , and CH 2 ;
W is absent, O, S or NR′;
V is a straight or branched, substituted or unsubstituted C 1 -C 24 alkylene, substituted or unsubstituted C 1 -C 24 alkenylene, or substituted or unsubstituted C 1 -C 24 alkynylene, of which one or more methylenes can be interrupted by O, S, S(O), SO 2 , N(R′), C(O), phosphorus containing linkage, aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic, or cycloalkyl;
R′ is hydrogen, acyl, unsubstituted or substituted C 1 -C 24 aliphatic;
Z is CONR 20 R 30 ;
R 20 and R 30 for each occurrence are independently selected from the group consisting of hydrogen, acyl, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic, OR 10 , COR 10 , CO 2 R 10 , and NR 10 R 10 ′; or R 20 and R 30 are taken together with the atom to which they are bound to form an unsubstituted or substituted 5-14 membered heterocyclic ring;
and
R 10 and R 10 ′ are each independently hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, heteroaryl, or heterocyclic,
wherein, in each occurrence, the substituent group is selected from the group consisting of halo, hydroxy, oxo, nitro, haloalkyl, alkyl, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, carboxyl, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, and ureido.
2. The nucleoside of claim 1 , represented by formula (3):
wherein:
one of D 1 and D 2 is H or a hydroxyl protecting group and the other of D 1 and D 2 is H, a hydroxyl protecting group, or a reactive phosphorus group, provided that when one of D 1 and D 2 is a reactive phosphorus group, the other of D 1 and D 2 is a hydroxyl protecting group;
B is selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, a natural nucleobase, and a universal base;
Q is selected from the group consisting of O, S, NR 10 , and CH 2 ;
W is absent, O, S or NR′;
R 100 and R 200 are each independently selected from the group consisting of hydrogen, unsubstituted and substituted C 1 -C 24 aliphatic, aryl, halogen, 5-14 membered heteroaryl, and 5-14 membered heterocyclic;
R 300 and R 400 are each independently selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, halogen, 5-14 membered heteroaryl, and 5-14 membered heterocyclic; or R 300 and R 400 are taken together with the atom to which they are bound to form a carbonyl, thiocarbonyl, ═NR 20 , ═N—NR 20 R 30 , or ═N—NR 20 C(O)R 10 ;
Z is CONR 20 R 30 ;
R 20 and R 30 are each independently selected from the group consisting of hydrogen, acyl, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic, OR 10 , COR 10 , CO 2 R 10 , and NR 10 R 10 ′; or R 20 and R 30 are taken together with the atom to which they are bound to form a 5-14 membered heterocyclic ring;
and
R 10 and R 10 ′ are each independently hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, or 5-14 membered heterocyclic.
3. The nucleoside of claim 1 , represented by formula (5):
wherein:
one of D 1 and D 2 is H, or a hydroxyl protecting group and the other of D 1 and D 2 is H, a hydroxyl protecting group, or a reactive phosphorus group, provided that when one of D 1 and D 2 is a reactive phosphorus group, the other of D 1 and D 2 is a hydroxyl protecting group;
B is selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, a natural nucleobase, and a universal base;
R 300 and R 400 are each independently selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, halogen, 5-14 membered heteroaryl, and 5-14 membered heterocyclic; or R 300 and R 400 are taken together with the atom to which they are bound to form a carbonyl, thiocarbonyl, ═NR 20 , ═N—NR 20 R 30 , or ═N—NR 20 C(O)R 10 ;
Z is CONR 20 R 30 ;
R 20 and R 30 for each occurrence are independently selected from the group consisting of hydrogen, acyl, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic, OR 10 , COR 10 , CO 2 R 10 , and NR 10 R 10 ′; or R 20 and R 30 are taken together with the atom to which they are bound to form a 5-14 membered heterocyclic ring;
and
R 10 and R 10 ′ are each independently hydrogen, an unsubstituted or substituted aliphatic, aryl, 5-14 membered heteroaryl, or 5-14 membered heterocyclic.
4. The nucleoside of claim 1 , represented by formula (7):
wherein:
one of D 1 and D 2 is H, or a hydroxyl protecting group and the other of D 1 and D 2 is H, a hydroxyl protecting group, or a reactive phosphorus group, provided that when one of D 1 and D 2 is a reactive phosphorus group, the other of D 1 and D 2 is a hydroxyl protecting group;
B is selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, a natural nucleobase, and a universal base;
R 100 and R 200 are each independently selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, halogen, 5-14 membered heteroaryl, and 5-14 membered heterocyclic; or R 100 and R 200 are taken together with the atom to which they are bound to form a carbonyl, thiocarbonyl, ═NR 20 , ═N—NR 20 R 30 , ═N—NR 20 C(O)R 10 or ═NR 20 ;
R 300 and R 400 are taken together with the atom to which they are bound to form a carbonyl;
R 500 -R 800 are each independently selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, halogen, 5-14 membered heteroaryl, and 5-14 membered heterocyclic;
Q 10 is O, S, or NR 20 ;
R 20 and R 30 are each independently selected from the group consisting of hydrogen, acyl, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic, OR 10 , COR 10 , CO 2 R 10 ,
and NR 10 R 10 ′; or R 20 and R 30 are taken together with the atom to which they are bound to form a 5-14 membered heterocyclic ring;
R 10 and R 10 ′ are independently hydrogen, C 1 -C 24 aliphatic, substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, or 5-14 membered heterocyclic;
s is 0, 1, 2, 3, 4, 5 or 6, and t is 1; and
u and v are independently for each occurrence 0, 1, 2, 3, 4, 5 or 6, provided that u and v are not both 0.
5. The nucleoside of claim 1 , represented by formula (9):
wherein:
one of D 1 and D 2 is H, or a hydroxyl protecting group and the other of D 1 and D 2 is H, a hydroxyl protecting group, or a reactive phosphorus group, provided that when one of D 1 and D 2 is a reactive phosphorus group, the other of D 1 and D 2 is a hydroxyl protecting group;
B is selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, a natural nucleobase, and a universal base;
R 100 and R 200 are each independently selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, halogen, 5-14 membered heteroaryl, and 5-14 membered heterocyclic; or R 100 and R 200 are taken together with the atom to which they are bound to form a carbonyl, thiocarbonyl, ═NR 20 , ═N—NR 20 R 30 , ═N—NR 20 C(O)R 10 or ═NR 20 ;
R 300 and R 400 are taken together with the atom to which they are bound to form a carbonyl;
Q 10 is O, S, or NR 20 ;
R 20 and R 30 are each independently selected from the group consisting of hydrogen, acyl, C 1 -C 24 aliphatic or substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic, OR 10 , COR 10 , CO 2 R 10 ,
and NR 10 R 10 ′; or R 20 and R 30 are taken together with the atom to which they are bound to form a 5-14 membered heterocyclic ring;
R 10 and R 10 ′ are independently hydrogen, C 1 -C 24 aliphatic, substituted aliphatic, aryl, 5-14 membered heteroaryl, or 5-14 membered heterocyclic; and
s is 0, 1, 2, 3, 4, 5 or 6, and t is 1.
6. The nucleoside of claim 1 , wherein the nucleoside has the formula of
wherein B is U, T, C Ac , A Bz , or G ibu .
7. An oligonucleotide comprising at least one nucleoside of formula (2):
wherein:
m is an integer from 1 to 15;
B is selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, a natural nucleobase, and a universal nucleobase;
Q is selected from the group consisting of O, S, NR 10 , and CH 2 ;
W is absent, O, S or NR′;
V is a straight or branched, substituted or unsubstituted C 1 -C 24 alkylene, substituted or unsubstituted C 1 -C 24 alkoxylene, substituted or unsubstituted C 1 -C 24 alkenylene, or substituted or unsubstituted C 1 -C 24 alkynylene, wherein the backbone of the alkylene, alkoxylene, alkenylene, or alkynylene may contain one or more of 0, S, S(O), SO 2 , N(R′), C(O), substituted or unsubstituted aryl, substituted or unsubstituted 5-14 membered heteroaryl, substituted or unsubstituted 5-14 membered heterocyclic, or substituted or unsubstituted cycloalkyl;
R′ is hydrogen, acyl, unsubstituted or substituted C 1 -C 24 aliphatic;
Z is CONR 20 R 30 ;
R 20 and R 30 for each occurrence are independently selected from the group consisting of hydrogen, acyl, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic, OR 10 , COR 10 , CO 2 R 10 , and NR 10 R 10 ′; or R 20 and R 30 are taken together with the atom to which they are bound to form an unsubstituted or substituted heterocyclic ring;
and
R 10 and R 10 ′ are each independently hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, or 5-14 membered heterocyclic,
wherein, in each occurrence, the substituent group is selected from the group consisting of halo, hydroxy, oxo, nitro, haloalkyl, alkyl, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, carboxyl, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, and ureido.
8. The oligonucleotide of claim 7 , wherein the nucleoside of formula (2) is represented by a nucleoside of formula (4):
wherein:
B is selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, a natural nucleobase, and a universal base;
Q is selected from the group consisting of O, S, NR 10 , and CH 2 ;
W is absent, O, S or NR′;
R 100 and R 200 are each independently selected from the group consisting of hydrogen, unsubstituted and substituted C 1 -C 24 aliphatic, aryl, halogen, 5-14 membered heteroaryl, and 5-14 membered heterocyclic;
R 300 and R 400 are each independently selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, halogen, 5-14 membered heteroaryl, and 5-14 membered heterocyclic; or R 300 and R 400 are taken together with the atom to which they are bound to form a carbonyl, thiocarbonyl, ═NR 20 , ═N—NR 20 R 30 , or ═N—NR 20 C(O)R 10 ;
Z is CONR 20 R 30 ;
R 20 and R 30 are independently selected from the group consisting of hydrogen, acyl, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic, OR 10 , COR 10 , CO 2 R 10 , and NR 10 R 10 ′; or R 20 and R 30 are taken together with the atom to which they are bound to form a heterocyclic ring;
and
R 10 and R 10 ′ are each independently hydrogen, unsubstituted or substituted aliphatic, aryl, 5-14 membered heteroaryl, or 5-14 membered heterocyclic.
9. The oligonucleotide of claim 7 , wherein the nucleoside of formula (2) is represented by a nucleoside of formula (6):
wherein:
B is selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, a natural nucleobase, and a universal base;
R 300 and R 400 are each independently selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, halogen, 5-14 membered heteroaryl, and 5-14 membered heterocyclic; or R 300 and R 400 are taken together with the atom to which they are bound to form a carbonyl, thiocarbonyl, ═NR 20 , ═N—NR 20 R 30 , or ═N—NR 20 C(O)R 10 ;
Z is CONR 20 R 30 ;
R 20 and R 30 for each occurrence are independently selected from the group consisting of hydrogen, acyl, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic, OR 10 , COR 10 , CO 2 R 10 , and NR 10 R 10 ′; or R 20 and R 30 are taken together with the atom to which they are bound to form a 5-14 membered heterocyclic ring;
and
R 10 and R 10 ′ are each independently hydrogen, an unsubstituted or substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, or 5-14 membered heterocyclic.
10. The oligonucleotide of claim 7 , wherein the nucleoside of formula (2) is represented by a nucleoside of formula (8):
wherein:
B is selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, a natural nucleobase, and a universal base;
R 100 and R 200 are each independently selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, halogen, 5-14 membered heteroaryl, and 5-14 membered heterocyclic; or R 100 and R 200 are taken together with the atom to which they are bound to form a carbonyl, thiocarbonyl, ═NR 20 , ═N—NR 20 R 30 , ═N—NR 20 C(O)R 10 or ═NR 20 ;
R 300 and R 400 are taken together with the atom to which they are bound to form a carbonyl;
R 500 -R 800 are each independently selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, halogen, 5-14 membered heteroaryl, and 5-14 membered heterocyclic;
Q 10 is O, S, or NR 20 ;
R 20 and R 30 are each independently selected from the group consisting of hydrogen, acyl, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic, OR 10 , COR 10 , CO 2 R 10 ,
and NR 10 R 10 ′; or R 20 and R 30 are taken together with the atom to which they are bound to form a 5-14 membered heterocyclic ring;
R 10 and R 10 ′ are independently hydrogen, C 1 -C 24 aliphatic, substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, or 5-14 membered heterocyclic;
s is 0, 1, 2, 3, 4, 5 or 6, and t is 1; and
u and v are independently for each occurrence 0, 1, 2, 3, 4, 5 or 6, provided that u and v are not both 0.
11. The oligonucleotide of claim 7 , wherein the nucleoside of formula (2) is represented by a nucleoside of formula (10):
wherein:
B is selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, a natural nucleobase, and a universal base;
R 100 and R 200 are each independently selected from the group consisting of hydrogen, unsubstituted or substituted C 1 -C 24 aliphatic, aryl, halogen, 5-14 membered heteroaryl, and 5-14 membered heterocyclic; or R 100 and R 200 are taken together with the atom to which they are bound to form a carbonyl, thiocarbonyl, ═NR 20 , ═N—NR 20 R 30 , ═N—NR 20 C(O)R 10 or ═NR 20 ;
R 300 and R 400 are taken together with the atom to which they are bound to form a carbonyl;
Q 10 is O, S, or NR 20 ;
R 20 and R 30 are each independently selected from the group consisting of hydrogen, acyl, C 1 -C 24 aliphatic or substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic, OR 10 , COR 10 , CO 2 R 10 ,
and NR 10 R 10 ′; or R 20 and R 30 are taken together with the atom to which they are bound to form a 5-14 membered heterocyclic ring;
R 10 and R 10 ′ are independently hydrogen, aliphatic, substituted C 1 -C 24 aliphatic, aryl, 5-14 membered heteroaryl, or 5-14 membered heterocyclic; and
s is 0, 1, 2, 3, 4, 5 or 6, and t is 1.
12. The oligonucleotide of claim 7 , wherein the nucleoside of formula (2) is selected from the group consisting of:
wherein:
R 1 , R 2 , R 3 , R 4 , R 6 , R 7 , R 8 and R 9 are each independently selected form the group consisting of alkyl, aryl, cycloalkyl, 5-14 membered heteroaryl, 5-14 membered heterocycloalkyl and acyl;
B is an optionally substituted natural or non-natural nucleobase or a universal nucleobase;
Q and Y are each independently selected from the group consisting of O, S, CR′ 2 , and NR″, where R′ is hydrogen, alkyl, aryl, cycloalkyl, 5-14 membered heteroaryl, or 5-14 membered heterocycloalkyl, and R″ is hydrogen, alkyl, aryl, cycloalkyl, 5-14 membered heteroaryl, 5-14 membered heterocycloalkyl or acyl; and
m, n and p are each independently an integer from 1 to 10.
13. The oligonucleotide of claim 7 , wherein said oligonucleotide further comprises at least one non-phosphodiester backbone linkage selected from the group consisting of phosphorothioate, phosphorodithioate, alkyl-phosphonate and phosphoramidate.
14. The oligonucleotide of claim 13 , wherein the non-phosphodiester backbone linkage is placed (a) between nucleosides of positions 1 and 2, and/or (b) between the 3′-terminal nucleosides.
15. The oligonucleotide of claim 7 , wherein said oligonucleotide further comprises at least one ligand conjugated to the nucleobases, sugar moieties, internucleosidic linkages, or the 3′ or 5′ end of the oligonucleotide.
16. The oligonucleotide of claim 7 , wherein said oligonucleotide is a double-stranded oligonucleotide comprising a first strand and a second strand.
17. The oligonucleotide of claim 7 , wherein the nucleoside of formula (2) is present in either the first strand or the second strand.
18. The oligonucleotide of claim 17 , wherein said first strand is the sense strand.
19. The oligonucleotide of claim 17 , wherein said second strand is the antisense strand.
20. The oligonucleotide of claim 17 , wherein the nucleoside of formula (2) is present in both strands.
21. The oligonucleotide of claim 7 , wherein said oligonucleotide has a hairpin structure.
22. The oligonucleotide of claim 7 , wherein the nucleoside has the formula of
wherein B is U, T, C Ac , A Bz , or G ibu .