Chimeric oligomeric compounds for modulation of splicing
View Patent ↗Disclosed herein are compounds, compositions and methods for modulating splicing of a selected target mRNA. Further provided are uses of the disclosed compounds and compositions in the manufacture of a medicament for treatment of diseases and disorders. Methods of enhancing cellular uptake, modulating tissue distribution and enhancing pharmacological activity of RNase H-independent antisense oligonucleotides are also provided.
1. A method of modulating splicing of a selected target pre-mRNA in a cell comprising contacting the cell with an oligomeric compound comprising an oligonucleotide consisting of 13 to 30 linked nucleosides which is complementary to the selected target pre-mRNA, and wherein the oligonucleotide has 7-11 separate regions, wherein:
the 5′-most region consists of 1-5 linked modified nucleosides, each comprising a modified sugar moiety;
the 3′-most region consists of 1-5 linked modified nucleosides, each comprising a modified sugar moiety; and
the internal regions alternate between regions of 1-5 linked 2′-deoxyribonucleosides and regions of 1-5 linked modified nucleosides, each comprising a modified sugar moiety,
wherein splicing of the selected target pre-mRNA is modulated.
2. The method of claim 1 , wherein said oligonucleotide comprises 3 separate regions of 2′-deoxyribonucleosides.
3. The method of claim 1 , wherein said oligonucleotide comprises 4 separate regions of 2′-deoxyribonucleosides.
4. The method of claim 1 , wherein each region of 2′-deoxyribonucleosides of said oligonucleotide consists of 2 or 3 nucleosides.
5. The method of claim 1 , wherein the 5′ most region of the oligonucleotide consists of 3 or 4 linked modified nucleosides each comprising a modified sugar moiety.
6. The method of claim 1 , wherein the 3′ most region of the oligonucleotide consists of 4 or 5 linked modified nucleosides each comprising a modified sugar moiety.
7. The method of claim 1 , wherein each sugar modification of each modified nucleoside is selected from the group consisting of 2′-O-methoxyethyl, 2′-fluoro, 2′-dimethylaminooxyethoxy, 2′-dimethylaminoethoxyethoxy, 2′-guanidinium, 2′-O-guanidinium ethyl, 2′-carbamate, 2′-aminooxy, 2′-acetamido and locked nucleic acid.
8. The method of claim 7 , wherein each sugar modification of each modified nucleoside is 2′-O-methoxyethyl.
9. The method of claim 1 , wherein the oligonucleotide consists of 20 to 30 linked nucleosides.
10. The method of claim 1 , wherein the oligonucleotide consists of 15 to 25 linked nucleosides.
11. The method of claim 1 , wherein the oligonucleotide consists of 20 linked nucleosides.
12. The method of claim 11 , wherein the oligonucleotide has 2′-deoxyribonucleosides at positions 3, 4, 5, 7, 8, 9, 11, 12, 13, 15, 16 and 17.
13. The method of claim 11 , wherein the oligonucleotide has 2′-deoxyribonucleosides at positions 4, 5, 6, 9, 10, 11, 14, 15 and 16.
14. The method of claim 11 , wherein the oligonucleotide has 2′-deoxyribonucleosides at positions 4, 5, 8, 9, 10, 13, 14 and 15.
15. The method of claim 11 , wherein the oligonucleotide has 2′-deoxyribonucleosides at positions 5, 6, 7, 9, 10, 11, 13, 14 and 15.
16. The method of claim 11 , wherein the oligonucleotide has 2′-deoxyribonucleosides at positions 5, 6, 7, 9, 10, 13, 14 and 15.
17. The method of claim 11 , wherein the oligonucleotide has 2′-deoxyribonucleosides at positions 5, 6, 7, 10, 11, 12, 15, 16 and 17.
18. The method of claim 11 , wherein the oligonucleotide has 2′-deoxyribonucleosides at positions 6, 7, 8, 10, 11, 12, 14, 15 and 16.
19. The method of claim 11 , wherein the oligonucleotide has 2′-deoxyribonucleosides at positions 6, 7, 8, 11, 12, 13, 16 and 17.
20. The method of claim 1 , wherein the cell is derived from adipose.
21. The method of claim 1 , wherein the cell is derived from liver.
22. The method of claim 1 , wherein the oligonucleotide is complementary to a splice site.
23. The method of claim 22 , wherein the splice site is an aberrant splice site.
24. The method of claim 22 , wherein the splice site is a cryptic splice site.