Oligomer-conjugate complexes and their use
Oligonucleotides, chemically-modified oligonucleotides, and oligonucleotide-conjugate complexes for use in research, diagnostics, and/or therapeutics are described herein. In some embodiments, oligonucleotides comprising a stabilized phosphate moiety covalently attached to the 5′-terminal nucleoside are provided.
1. An oligomeric compound comprising an oligonucleotide consisting of 10-30 linked nucleosides and at least one conjugate group, wherein the oligomeric compound comprises a phosphonate moiety covalently attached to the 5′-terminal nucleoside; and wherein the phosphonate moiety comprises the following formula:
wherein:
R a and R c are each, independently, OH, SH, C 1 -C 6 alkyl, substituted C 1 -C 6 alkyl, C 1 -C 6 alkoxy, substituted C 1 -C 6 alkoxy, amino or substituted amino;
R b is O or S; and
X is C(R 1 )(R 2 ) wherein R 1 and R 2 are independently selected from: H, halogen, OH, SH, C 1 -C 6 alkyl, substituted C 1 -C 6 alkyl, C 1 -C 6 alkoxy, substituted C 1 -C 6 alkoxy, amino or substituted amino.
2. The oligomeric compound of claim 1 , wherein the phosphonate moiety comprises the following formula:
wherein:
R a and R c are each, independently, OH, SH, C 1 -C 6 alkyl, substituted C 1 -C 6 alkyl, C 1 -C 6 alkoxy, substituted C 1 -C 6 alkoxy, amino or substituted amino;
R b is O or S;
X is C(R 1 )(R 2 ) and;
Y is selected from C(R 3 )(R 4 ), S, and N; wherein
R 1 and R 3 are independently selected from: H, halogen, OH, SH, C 1 -C 6 alkyl, substituted C 1 -C 6 alkyl, C 1 -C 6 alkoxy, substituted C 1 -C 6 alkoxy, amino or substituted amino or R 1 and R 3 together form a bond; and
R 2 and R 4 are independently selected from: H, halogen, OH, SH, C 1 -C 6 alkyl, substituted C 1 -C 6 alkyl, C 1 -C 6 alkoxy, substituted C 1 -C 6 alkoxy, amino or substituted amino or R 2 and R 4 together form a bond.
3. The oligomeric compound of claim 1 , wherein the conjugate group is covalently attached to the oligonucleotide at a nucleoside at position 1, 2, 3, 4, 6, 7, 8, 9, 18, 19, 20, or 21 from the 5′-end of the oligonucleotide or at position 1, 2, 3, 12, 13, 4, 15, 17, 18, 19, 20, or 21 from the 3′-end of the oligonucleotide.
4. The oligomeric compound of claim 1 , wherein the conjugate group is covalently attached to any of the 1 to 4 5′-most nucleosides of the oligonucleotide.
5. The oligomeric compound of claim 1 , wherein the conjugate group is covalently attached to the 5′-terminal nucleoside of the oligonucleotide.
6. The oligomeric compound of claim 1 , wherein the conjugate group is covalently attached to the 8 th nucleoside from the 5′−terminal end of the oligonucleotide.
7. The oligomeric compound of claim 1 , wherein the conjugate group is covalently attached to the 6 th nucleoside from the 5′−terminal end of the oligonucleotide.
8. The oligomeric compound of claim 1 , wherein the conjugate group comprises a C 10 -C 20 alkyl.
9. The oligomeric compound of claim 1 , wherein the conjugate group comprises a C 10 -C 20 alkenyl.
10. The oligomeric compound of claim 1 , wherein the conjugate group comprises a steroid.
11. The oligomeric compound of claim 1 , wherein the conjugate group comprises cholesterol.
12. The oligomeric compound of claim 1 , wherein the conjugate group comprises a carbohydrate.
13. The oligomeric compound of claim 1 , wherein the conjugate group comprises N-acetylgalactosamine.
14. The oligomeric compound of claim 1 , wherein the oligonucleotide consists of 19 linked nucleosides.
15. The oligomeric compound of claim 1 , wherein the oligonucleotide consists of 20 linked nucleosides.
16. The oligomeric compound of claim 1 , wherein the oligonucleotide consists of 21 linked nucleosides.
17. The oligomeric compound of claim 1 , wherein the oligonucleotide comprises at least one 2′-modified nucleoside.
18. The oligomeric compound of claim 1 , wherein the oligomeric compound is single-stranded.
19. The compound of claim 1 , wherein the oligomeric compound is double-stranded.
20. A method of inhibiting protein expression in a cell comprising contacting the cell with the oligomeric compound according to claim 1 .