Modulators of DNM2 expression
The present embodiments provide methods, compounds, and compositions useful for inhibiting DNM2 expression, which may be useful for treating, preventing, or ameliorating a disease associated with DNM2.
1. A compound comprising a modified oligonucleotide consisting of 16 to 20 linked nucleosides and having a nucleobase sequence comprising at least 14 contiguous nucleobases of SEQ ID NO: 2879 or SEQ ID NO: 2123, wherein the modified oligonucleotide comprises at least one of a modified internucleoside linkage and a modified sugar moiety.
2. The compound of claim 1 , wherein the nucleobase sequence comprises SEQ ID NO: 2879.
3. The compound of claim 1 , wherein the nucleobase sequence comprises SEQ ID NO: 2123.
4. The compound of claim 1 , wherein the nucleobase sequence is SEQ ID NO: 2879 or 2123.
5. The compound of claim 1 , wherein the modified oligonucleotide comprises:
a gap segment consisting of 8-12 linked 2′-deoxynucleosides;
a 5′ wing segment consisting of 1-7 linked nucleosides; and
a 3′ wing segment consisting of 1-7 linked nucleosides;
wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein the terminal nucleoside of each wing segment comprises a modified sugar.
6. The compound of claim 1 , wherein the modified oligonucleotide comprises:
a gap segment consisting of 10 linked 2′-deoxynucleosides;
a 5′ wing segment consisting of 3 linked nucleosides; and
a 3′ wing segment consisting of 3 linked nucleosides;
wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of each wing segment comprises a cEt sugar moiety; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine.
7. The compound of claim 1 , wherein the modified internucleoside linkage is a phosphorothioate internucleoside linkage.
8. The compound of claim 1 , wherein the modified sugar moiety is a bicyclic sugar.
9. The compound of claim 8 , wherein the bicyclic sugar is selected from the group consisting of LNA, ENA, and cEt.
10. The compound of claim 1 , wherein the modified oligonucleotide comprises at least one 5-methylcytosine.
11. The compound of claim 1 , wherein the modified oligonucleotide is single-stranded.
12. The compound of claim 1 , wherein the compound is a double-stranded compound.
13. The compound of claim 1 , comprising a conjugate group.
14. The compound of claim 13 , wherein the compound consists of the modified oligonucleotide and the conjugate group.
15. A modified oligonucleotide according to the following formula: Gks Tks Tks Tds Ads Tds Tds Ads Tds Ads Gds Gds Gds mCks Tks Tk; wherein,
A=an adenine nucleobase,
mC=a 5-methylcytosine nucleobase,
G=a guanine nucleobase,
T=a thymine nucleobase,
k=a cEt sugar moiety,
d=a 2′-deoxyribosyl sugar moiety, and
s=a phosphorothioate internucleoside linkage.
16. A modified oligonucleotide, wherein the anion form of the modified oligonucleotide has the following chemical structure:
17. The modified oligonucleotide of claim 16 , wherein the modified oligonucleotide is a salt.
18. The modified oligonucleotide of claim 17 , wherein the cation of the salt is a sodium or a potassium.
19. A chirally enriched population of the compound of claim 1 , wherein the population is enriched for compounds having a modified oligonucleotide comprising at least one particular phosphorothioate internucleoside linkage having a particular stereochemical configuration.
20. A pharmaceutical composition comprising the modified oligonucleotide of claim 16 , and at least one pharmaceutically acceptable diluent or carrier.
21. A method of treating, preventing, or ameliorating centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease, the method comprising administering the compound of claim 1 to a subject with a mutation in at least one gene selected from among MTM1, BIN1, and DNM2.
22. An in vitro method of inhibiting expression of DNM2 in a cell comprising contacting the cell with compound comprising a single-stranded modified oligonucleotide that is 100% complementary to exon 10, intron 1, intron 11, intron 12, intron 13, intron 14, or the 3′-UTR of a DNM2 nucleic acid transcript, thereby inhibiting expression of DNM2 in the cell.
23. The in vitro method of claim 22 , wherein the modified oligonucleotide consists of 16 to 20 linked nucleosides and has a nucleobase sequence comprising at least 14 contiguous nucleobases of SEQ ID NO: 2879 or SEQ ID NO: 2123, wherein the modified oligonucleotide comprises at least one of a modified internucleoside linkage and a modified sugar moiety.
24. The in vitro method of claim 22 , wherein the modified oligonucleotide is according to the following formula: Gks Tks Tks Tds Ads Tds Tds Ads Tds Ads Gds Gds Gds mCks Tks Tk; wherein,
A=an adenine nucleobase,
mC=a 5-methylcytosine nucleobase,
G=a guanine nucleobase,
T=a thymine nucleobase,
k=a cEt sugar moiety,
d=a 2′-deoxyribosyl sugar moiety, and
s=a phosphorothioate internucleoside linkage.
25. The in vitro method of claim 22 , wherein the anion form of the modified oligonucleotide has the following chemical structure:
26. The in vitro method of claim 22 , wherein the modified oligonucleotide has the following chemical structure:
or a salt thereof.
27. The in vitro method of claim 22 , wherein the modified oligonucleotide has the following chemical structure:
28. The pharmaceutical composition of claim 20 , wherein the pharmaceutically acceptable diluent is phosphate buffered saline.
29. A chirally enriched population of the modified oligonucleotide of claim 15 , wherein the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate internucleoside linkage having a particular stereochemical configuration.
30. A chirally enriched population of the modified oligonucleotide of claim 16 , wherein the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate internucleoside linkage having a particular stereochemical configuration.
31. A pharmaceutical composition comprising the modified oligonucleotide of claim 15 , and at least one pharmaceutically acceptable diluent or carrier.
32. The pharmaceutical composition of claim 31 , wherein the pharmaceutically acceptable diluent is phosphate buffered saline.
33. A method of treating or ameliorating centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease, the method comprising administering the modified oligonucleotide of claim 15 to a subject with a mutation in at least one gene selected from among MTM1, BIN1, and DNM2.
34. A method of treating or ameliorating centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease, the method comprising administering the modified oligonucleotide of claim 16 to a subject with a mutation in at least one gene selected from among MTM1, BIN1, and DNM2.
35. The modified oligonucleotide of claim 15 , comprising the modified oligonucleotide covalently linked to a conjugate group.
36. A modified oligonucleotide according to the following chemical structure:
or a salt thereof.
37. The modified oligonucleotide of claim 36 , which is the sodium salt or the potassium salt.
38. A chirally enriched population of the modified oligonucleotide of claim 36 , wherein the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate internucleoside linkage having a particular stereochemical configuration.
39. A pharmaceutical composition comprising the modified oligonucleotide of claim 36 and at least one pharmaceutically acceptable diluent or carrier.
40. The pharmaceutical composition of claim 39 , wherein the pharmaceutically acceptable diluent is phosphate buffered saline.
41. A method of treating or ameliorating centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease, the method comprising administering the modified oligonucleotide of claim 36 to a subject with a mutation in at least one gene selected from among MTM1, BIN1, and DNM2.
42. A modified oligonucleotide according to the following chemical structure:
43. A chirally enriched population of the modified oligonucleotide of claim 42 , wherein the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate internucleoside linkage having a particular stereochemical configuration.
44. A pharmaceutical composition comprising the modified oligonucleotide of claim 42 and at least one pharmaceutically acceptable diluent or carrier.
45. The pharmaceutical composition of claim 44 , wherein the pharmaceutically acceptable diluent is phosphate buffered saline.
46. A method of treating or ameliorating centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease, the method comprising administering the modified oligonucleotide of claim 42 to a subject with a mutation in at least one gene selected from among MTM1, BIN1, and DNM2.
47. The pharmaceutical composition of claim 32 , consisting essentially of the modified oligonucleotide and phosphate buffered saline.
48. The pharmaceutical composition of claim 40 , consisting essentially of the modified oligonucleotide and phosphate buffered saline.
49. The pharmaceutical composition of claim 45 , consisting essentially of the modified oligonucleotide and phosphate buffered saline.