MicroRNA compounds and methods for modulating miR-122
Described herein are compositions and methods for the inhibition of miR-122 activity. The compositions have certain nucleoside modifications that yield potent inhibitors of miR-122 activity. The compounds may comprise conjugates to facilitate delivery to the liver. The compositions may be administered to subjects infected with hepatitis C virus, as a treatment for hepatitis C virus and related conditions.
1. A compound comprising a modified oligonucleotide having the structure:
A E Me C E A E Me C E Me C E A E T E TGU S C S AC S AC S TC S C S (SEQ ID NO: 4)
wherein the superscript “Me” indicates 5-methylcytosine; nucleosides not followed by a subscript are β-D-deoxyribonucleosides; nucleosides followed by a subscript “E” are 2′-MOE nucleosides; nucleosides followed by a subscript “S” are S-cEt nucleosides; and each internucleoside linkage is a phosphorothioate internucleoside linkage.
2. The compound of claim 1 , wherein the compound comprises a conjugate moiety linked to the 5′ terminus or the 3′ terminus of the modified oligonucleotide.
3. The compound of claim 2 , wherein the conjugate moiety comprises at least one ligand selected from a carbohydrate, cholesterol, a lipid, a phospholipid, an antibody, a lipoprotein, a hormone, a peptide, a vitamin, a steroid, and a cationic lipid.
4. The compound of claim 2 , wherein the compound has the structure:
L n -linker-MO
wherein each L is, independently, a ligand and n is from 1 to 10; and MO is the modified oligonucleotide.
5. The compound of claim 2 , wherein the compound has the structure:
L n -linker-X-MO
wherein each L is, independently, a ligand and n is from 1 to 10; X is a phosphodiester linkage or a phosphorothioate linkage; and MO is the modified oligonucleotide.
6. The compound of claim 2 , wherein the compound has the structure:
L n -linker-X 1 -N m -X 2 -MO
wherein each L is, independently, a ligand and n is from 1 to 10; each N in N m is, independently, a modified or unmodified nucleoside and m is from 1 to 5; X 1 and X 2 are each, independently, a phosphodiester linkage or a phosphorothioate linkage; and MO is the modified oligonucleotide.
7. The compound of claim 6 , wherein the compound has the structure:
wherein:
B is selected from —O—, —S—, —N(R N )—, —Z—P(Z′)(Z″)O—, —Z—P(Z′)(Z″)O—N m —X—, and —Z—P(Z′)(Z″)O—N m —Y—;
MO is the modified oligonucleotide;
R N is selected from H, methyl, ethyl, propyl, isopropyl, butyl, and benzyl;
Z, Z′, and Z″ are each independently selected from O and S;
each N in N m is, independently, a modified or unmodified nucleoside;
m is from 1 to 5;
X is selected from a phosphodiester linkage and a phosphorothioate linkage;
Y is a phosphodiester linkage; and
the wavy line indicates the connection to the rest of the linker and ligand(s).
8. The compound of claim 6 wherein n is from 1 to 5.
9. The compound of claim 6 , wherein at least one ligand is a carbohydrate.
10. The compound of claim 6 , wherein at least one ligand is selected from mannose, glucose, galactose, ribose, arabinose, fructose, fucose, xylose, D-mannose, L-mannose, D-galactose, L-galactose, D-glucose, L-glucose, D-ribose, L-ribose, D-arabinose, L-arabinose, D-fructose, L-fructose, D-fucose, L-fucose, D-xylose, L-xylose, alpha-D-mannofuranose, beta-D-mannofuranose, alpha-D-mannopyranose, beta-D-mannopyranose, alpha-D-glucofuranose, Beta-D-glucofuranose, alpha-D-glucopyranose, beta-D-glucopyranose, alpha-D-galactofuranose, beta-D-galactofuranose, alpha-D-galactopyranose, beta-D-galactopyranose, alpha-D-ribofuranose, beta-D-ribofuranose, alpha-D-ribopyranose, beta-D-ribopyranose, alpha-D-fructofuranose, alpha-D-fructopyranose, glucosamine, galactosamine, sialic acid, and N-acetylgalactosamine.
11. The compound of claim 6 , wherein each ligand is N-acetylgalactosamine.
12. The compound of claim 6 , wherein the compound has the structure:
wherein each N in N m is, independently, a modified or unmodified nucleoside and m is from 1 to 5; X 1 and X 2 are each, independently, a phosphodiester linkage or a phosphorothioate linkage; and MO is the modified oligonucleotide.
13. The compound of claim 12 , wherein at least one of X 1 and X 2 is a phosphodiester linkage.
14. The compound of claim 6 , wherein N m is N′ p N″, wherein each N′ is, independently, a modified or unmodified nucleoside and p is from 0 to 4; and N″ is a nucleoside comprising an unmodified sugar moiety.
15. The compound of claim 14 , wherein each N′ comprises an unmodified sugar moiety.
16. The compound of claim 14 , wherein N″ is a β-D-deoxyriboadenosine or a β-D-deoxyriboguanosine.
17. The compound of claim 6 , wherein the sugar moiety of each N in N m is independently selected from a β-D-ribose, a β-D-deoxyribose, a 2′-O-methoxy sugar, a 2′-O-methyl sugar, a 2′-fluoro sugar, and a bicyclic sugar moiety.
18. A conjugate having the structure
wherein X is a phosphodiester linkage; m is 1; N in N m is a β-D-deoxyriboadenosine; Y is a phosphodiester linkage; and MO is a modified oligonucleotide having the structure A E Me C E A E Me C E Me C E A E T E TGU S C S AC S TC S C S (SEQ ID NO: 4), wherein the superscript “Me” indicates 5-methylcytosine, wherein nucleosides not followed by a subscript are β-D-deoxyribonucleosides, nucleosides followed by a subscript “E” are 2′-MOE nucleosides, nucleosides followed by a subscript “S” are S-cEt nucleosides, and each internucleoside linkage is a phosphorothioate internucleoside linkage; and wherein Y is linked to the 3′ terminus of the modified oligonucleotide.
19. A pharmaceutical composition comprising a conjugate having the structure:
wherein X is a phosphodiester linkage; m is 1; N in N m is a β-D-deoxyriboadenosine; Y is a phosphodiester linkage; and MO is a modified oligonucleotide having the structure A E Me C E A E Me C E Me C E A E T E TGU S C S AC S AC S TC S C S (SEQ ID NO: 4), wherein the superscript “Me” indicates 5-methylcytosine, wherein nucleosides not followed by a subscript are β-D-deoxyribonucleosides, nucleosides followed by a subscript “E” are 2′-MOE nucleosides, nucleosides followed by a subscript “S” are S-cEt nucleosides, and each internucleoside linkage is a phosphorothioate internucleoside linkage; and wherein Y is linked to the 3′ terminus of the modified oligonucleotide;
and one or more pharmaceutically acceptable excipients.
20. The pharmaceutical composition of claim 19 , which is a sterile aqueous solution.