Compounds and methods for modulating gene expression
The present disclosure describes short antisense compounds, including such compounds comprising chemically-modified high-affinity monomers 8-16 monomers in length. Certain such short antisense compound are useful for the reduction of target nucleic acids and/or proteins in cells, tissues, and animals with increased potency and improved therapeutic index. Thus, provided herein are short antisense compounds comprising high-affinity nucleotide modifications useful for reducing a target RNA in vivo. Such short antisense compounds are effective at lower doses than previously described antisense compounds, allowing for a reduction in toxicity and cost of treatment. In addition, the described short antisense compounds have greater potential for oral dosing.
1. A compound comprising a short antisense oligonucleotide consisting of 12 to 14 linked nucleosides and having:
a gap region consisting of 8 to 12 linked 2′-deoxynucleosides;
a 5′-wing region adjacent to the gap region at the 5′side of the gap region and consisting of 1 or 2 linked modified nucleosides; and
a 3′-wing region adjacent to the gap region at the 3′ side of the gap region and consisting of 1 or 2 linked modified nucleosides; wherein
at least one modified nucleoside of one or both of the 5′-wing region and the 3′-wing region is a high-affinity modified monomer.
2. The compound of claim 1 , wherein the short antisense oligonucleotide consists of 12 linked nucleosides.
3. The compound of claim 1 , wherein the short antisense oligonucleotide consists of 13 linked nucleosides.
4. The compound of claim 1 , wherein the short antisense oligonucleotide consists of 14 linked nucleosides.
5. The compound of claim 1 , wherein the gap region consists of 8 linked 2′-deoxynucleosides.
6. The compound of claim 1 , wherein the gap region consists of 9 linked 2′-deoxynucleosides.
7. The compound of claim 1 , wherein the gap region consists of 10 linked 2′-deoxynucleosides.
8. The compound of claim 1 , wherein the gap region consists of 11 linked 2′-deoxynucleosides.
9. The compound of claim 1 , wherein the gap region consists of 12 linked 2′-deoxynucleosides.
10. The compound of claim 1 , wherein the short antisense oligonucleotide has a motif selected from 1-10-1; 2-10-1; 1-10-2; 2-10-2; and 2-8-2 wherein, the first number represents the number of modified nucleosides in the 5′-wing region, the second number represents the number of linked 2′-deoxynucleosides in the gap region, and the third number represents the number of modified nucleosides in the 3′-wing region.
11. The compound of claim 10 , wherein the motif is selected from 1-10-1; 2-10-2; and 2-8-2.
12. The compound of claim 1 , wherein the short antisense compound comprises at least one modified internucleoside linkage.
13. The compound claim 12 , wherein the short antisense compound comprises at least one phosphorothioate linkage.
14. The compound of claim 12 , wherein each internucleoside linkage of the short antisense oligonucleotide is a modified internucleoside linkage.
15. The compound claim 14 , wherein each modified internucleoside linkage is a phosphorothioate linkage.
16. The compound of claim 1 , wherein each modified nucleoside of one or both of the 5′-wing region and the 3′-wing region comprises a 2′-O(CH 2 ) 2 OCH 3 substituent.
17. The compound of claim 1 , wherein each modified nucleoside of both the 5′-wing region and the 3′-wing region comprises a 2′-O(CH 2 ) 2 OCH 3 substituent.
18. The compound of claim 1 , wherein each high-affinity modified monomer is a bicyclic nucleotide.
19. The compound of claim 18 , wherein each bicyclic nucleotide comprises a 4′-CH2-O-2′ bridge.
20. The compound of claim 1 , wherein each high-affinity modified monomer is a bicyclic nucleotide.