Antisense conjugates for decreasing expression of DMPK
The disclosure provides novel conjugates comprising antisense oligonucleotides that hybridize to a DMPK transcript and a 3E10 antibody or binding fragment thereof. Also considered are these conjugates further comprising MBNL1 polypeptides. Methods of treating myotonic dystrophy using these conjugates and kits comprising these conjugates are also considered. Wherein the conjugates are suitable for delivery to muscle cells.
1. A conjugate comprising
an antisense oligonucleotide that hybridizes to a DMPK transcript; and
an antibody or antigen binding fragment, wherein the antibody or antigen binding fragment comprises a heavy chain variable domain comprising an amino acid sequence at least 95% identical to SEQ ID NO: 13, or wherein the antibody or antigen binding fragment comprises a light chain variable domain comprising an amino acid sequence at least 95% identical to SEQ ID NO: 14.
2. The conjugate claim 1 , wherein the antibody or antigen binding fragment is a chimeric, humanized, or fully human antibody or antigen binding fragment.
3. The conjugate of claim 1 , wherein the antibody or antigen binding fragment comprises a heavy chain variable domain comprising an amino acid sequence at least 95% identical to SEQ ID NO: 13 and a light chain variable domain comprising an amino acid sequence at least 95% identical to SEQ ID NO: 14, or a humanized variant thereof.
4. The conjugate of claim 1 , wherein the antibody or antigen binding fragment comprises
a VH CDR1 having the amino acid sequence of SEQ ID NO: 16;
a VH CDR2 having the amino acid sequence of SEQ ID NO: 17;
a VH CDR3 having the amino acid sequence of SEQ ID NO: 18;
a VL CDR1 having the amino acid sequence of SEQ ID NO: 19;
a VL CDR2 having the amino acid sequence of SEQ ID NO: 20; and
a VL CDR3 having the amino acid sequence of SEQ ID NO: 21.
5. The conjugate of claim 1 , wherein the antisense oligonucleotide hybridizes to CUG repeats to at least a portion of the 3′ UTR of a DMPK transcript.
6. The conjugate of claim 1 , wherein the antisense oligonucleotide hybridizes to any one of SEQ ID NOs: 24-27.
7. The conjugate of claim 1 , wherein the antisense oligonucleotide is capable, upon hybridization to its target sequence, of mediating RNase H-mediated degradation.
8. The conjugate of claim 1 , wherein the antisense oligonucleotide prevents proteins or nucleic acids from binding to at least a portion of the 3′UTR of a DMPK transcript.
9. The conjugate of claim 1 , wherein the antisense oligonucleotide prevents a protein or nucleic acid from binding to any one of SEQ ID NOs: 24-27.
10. The conjugate of claim 1 , wherein the antisense oligonucleotide comprises two wing portions, each of which flank the central portion.
11. The conjugate of claim 10 , wherein each of the two wing portions comprises at least 3 nucleotides.
12. The conjugate of claim 10 , wherein each of the two wing portions is of differing length.
13. The conjugate of claim 10 , wherein each of the two wing portions comprises at least 3 nucleotides modified to increase stability or specificity of the oligonucleotide.
14. The conjugate of claim 13 , wherein each of the two wing portions comprises unmodified and/or modified nucleotides.
15. The conjugate of claim 1 , wherein the ratio of antisense oligonucleotide conjugated per antibody or antigen binding fragment is 1:1.
16. The conjugate of claim 1 , wherein the conjugate is capable of penetrating muscle cells.
17. The conjugate of claim 1 , wherein the conjugate further comprises a muscle blind polypeptide (MBNL1), or a functional fragment thereof comprising all four zinc finger motifs, interconnected to the antibody or antigen fragment thereof.
18. A method of treating myotonic dystrophy, comprising administering to a patient in need thereof a conjugate of claim 1 .
19. A method of promoting entry into muscle cells, comprising contacting cells or administering to a patient a conjugate of claim 1 .
20. A conjugate comprising:
an antisense oligonucleotide that hybridizes to a DMPK transcript; and
an antibody or antigen binding fragment, wherein the antibody or antigen binding fragment comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 13 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 14, or a humanized variant thereof.
21. The conjugate of claim 20 , wherein the antibody or antigen binding fragment is a chimeric, humanized, or fully human antibody or antigen binding fragment.
22. The conjugate of claim 20 , wherein the antibody or antigen binding fragment comprises
a VH CDR1 having the amino acid sequence of SEQ ID NO: 16;
a VH CDR2 having the amino acid sequence of SEQ ID NO: 17;
a VH CDR3 having the amino acid sequence of SEQ ID NO: 18;
a VL CDR1 having the amino acid sequence of SEQ ID NO: 19;
a VL CDR2 having the amino acid sequence of SEQ ID NO: 20; and
a VL CDR3 having the amino acid sequence of SEQ ID NO: 21.
23. The conjugate of claim 20 , wherein the antisense oligonucleotide hybridizes to CUG repeats to at least a portion of the 3′ UTR of a DMPK transcript.
24. The conjugate of claim 20 , wherein the antisense oligonucleotide hybridizes to any one of SEQ ID NOs: 24-27.
25. The conjugate of claim 20 , wherein the antisense oligonucleotide is capable, upon hybridization to its target sequence, of mediating RNase H-mediated degradation.
26. The conjugate of claim 20 , wherein the antisense oligonucleotide prevents proteins or nucleic acids from binding to at least a portion of the 3′UTR of a DMPK transcript.
27. The conjugate of claim 20 , wherein the antisense oligonucleotide prevents a protein or nucleic acid from binding to any one of SEQ ID NOs: 24-27.
28. The conjugate of claim 20 , wherein the antisense oligonucleotide comprises two wing portions, each of which flank the central portion.
29. The conjugate of claim 28 , wherein each of the two wing portions comprises at least 3 nucleotides.
30. The conjugate of claim 28 , wherein each of the two wing portions is of differing length.
31. The conjugate of claim 28 , wherein each of the two wing portions comprises at least 3 nucleotides modified to increase stability or specificity of the oligonucleotide.
32. The conjugate of claim 31 , wherein each of the two wing portions comprises unmodified and/or modified nucleotides.
33. The conjugate of claim 20 , wherein the ratio of antisense oligonucleotide conjugated per antibody or antigen binding fragment is 1:1.
34. The conjugate of claim 20 , wherein the conjugate is capable of penetrating muscle cells.
35. The conjugate of claim 20 , wherein the conjugate further comprises a muscle blind polypeptide (MBNL1), or a functional fragment thereof comprising all four zinc finger motifs, interconnected to the antibody or antigen fragment thereof.
36. A method of treating myotonic dystrophy, comprising administering to a patient in need thereof a conjugate of claim 20 .
37. A method of promoting entry into muscle cells, comprising contacting cells or administering to a patient a conjugate of claim 20 .
38. A conjugate comprising
an antisense oligonucleotide that hybridizes to a DMPK transcript; and
an antibody or antigen binding fragment, wherein the antibody or antigen binding fragment comprises
a VH CDR1 having the amino acid sequence of SEQ ID NO: 16;
a VH CDR2 having the amino acid sequence of SEQ ID NO: 17;
a VH CDR3 having the amino acid sequence of SEQ ID NO: 18;
a VL CDR1 having the amino acid sequence of SEQ ID NO: 19;
a VL CDR2 having the amino acid sequence of SEQ ID NO: 20; and
a VL CDR3 having the amino acid sequence of SEQ ID NO: 21.
39. The conjugate of claim 38 , wherein the antibody or antigen binding fragment comprises a heavy chain variable domain comprising an amino acid sequence at least 95% identical to SEQ ID NO: 13, or wherein the antibody or antigen binding fragment comprises a light chain variable domain comprising an amino acid sequence at least 95% identical to SEQ ID NO: 14.
40. The conjugate of claim 38 , wherein the antibody or antigen binding fragment comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 13 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 14, or a humanized variant thereof.
41. The conjugate of claim 38 , wherein the antibody or antigen binding fragment is a chimeric, humanized, or fully human antibody or antigen binding fragment.
42. The conjugate of claim 38 , wherein the antisense oligonucleotide hybridizes to CUG repeats to at least a portion of the 3′ UTR of a DMPK transcript.
43. The conjugate of claim 38 , wherein the antisense oligonucleotide hybridizes to any one of SEQ ID NOs: 24-27.
44. The conjugate of claim 38 , wherein the antisense oligonucleotide is capable, upon hybridization to its target sequence, of mediating RNase H-mediated degradation.
45. The conjugate of claim 38 , wherein the antisense oligonucleotide prevents proteins or nucleic acids from binding to at least a portion of the 3′UTR of a DMPK transcript.
46. The conjugate of claim 38 , wherein the antisense oligonucleotide prevents a protein or nucleic acid from binding to any one of SEQ ID NOs: 24-27.
47. The conjugate of claim 38 , wherein the antisense oligonucleotide comprises two wing portions, each of which flank the central portion.
48. The conjugate of claim 47 , wherein each of the two wing portions comprises at least 3 nucleotides.
49. The conjugate of claim 47 , wherein each of the two wing portions is of differing length.
50. The conjugate of claim 47 , wherein each of the two wing portions comprises at least 3 nucleotides modified to increase stability or specificity of the oligonucleotide.
51. The conjugate of claim 50 , wherein each of the two wing portions comprises unmodified and/or modified nucleotides.
52. The conjugate of claim 38 , wherein the ratio of antisense oligonucleotide conjugated per antibody or antigen binding fragment is 1:1.
53. The conjugate of claim 38 , wherein the conjugate is capable of penetrating muscle cells.
54. The conjugate of claim 38 , wherein the conjugate further comprises a muscle blind polypeptide (MBNL1), or a functional fragment thereof comprising all four zinc finger motifs, interconnected to the antibody or antigen fragment thereof.
55. A method of treating myotonic dystrophy, comprising administering to a patient in need thereof a conjugate of claim 38 .
56. A method of promoting entry into muscle cells, comprising contacting cells or administering to a patient a conjugate of claim 38 .