Chemically modified oligonucleotides for use in modulation micro RNA and uses thereof
View Patent ↗This invention relates generally to chemically modified oligonuceotides useful for modulating expression of microRNAs and pre-microRNAs. More particularly, the invention relates to single stranded chemically modified oligonuceotides for inhibiting microRNA and pre-microRNA expression and to methods of making and using the modified oligonucleotides. Also included in the invention are compositions and methods for silencing microRNAs in the central nervous system.
1. A method of reducing plasma cholesterol levels in a mammal, the method comprising administering an antagomir to the mammal, wherein the antagomir is 15 to 25 nucleotides in length and has no more than two mismatches to the sequence ofmiR-122 (SEQ ID NO: 1), further wherein said antagomir comprises a non-nucleotide moiety at the 3′ -end, a 2′ -modification at each nucleotide, a phosphorothioate at the first and second internucleotide linkages at the 5′ end of the nucleotide sequence, a phosphorothioate at each of the first three internucleotide linkages at the 3′ end of the nucleotide sequence, and a phosphorothioate linkage between the 3′ terminal nucleotide and the non-nucleotide moiety, and wherein the remaining internucleotide linkages are phosphodiester.
2. The method of claim 1 , wherein the non-nucleotide moiety is a cholesterol moiety.
3. The method of claim 1 , wherein the 2′-modified nucleotide comprises a modification selected from the group consisting of: 2′-deoxy, 2′-deoxy-2′-fluoro,
2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), and 2′-O——N-methylacetamido (2′-O-NMA).
4. The method of claim 3 , wherein the 2′-modified nucleotide comprises a 2′-O-methyl.
5. The method of claim 1 , wherein the antagomir has no more than 1 mismatches to the target sequence.
6. The method of claim 1 , wherein the antagomir has no mismatches to the target sequence.
7. The method of claim 1 , wherein the antagomir is 18 to 25 nucleotides in length.
8. The method of claim 1 , wherein the antagomir is present in a pharmaceutical composition.
9. The method of claim 1 , wherein the administering comprises parenteral administration.
10. The method of claim 1 , wherein the administering comprises intravenous administration.
11. The method of claim 1 , further comprising reducing HMG-CoA reductase activity.
12. The method of claim 1 , wherein the antagomir has the sequence of SEQ ID NO: 5, and the non-nucleotide moiety is a cholesterol moiety.