Compositions and methods of treating muscle atrophy and myotonic dystrophy
Disclosed herein are polynucleic acid molecules, pharmaceutical compositions, and methods for treating muscle atrophy or myotonic dystrophy.
1. A single-stranded antisense oligonucleotide (ASO) conjugate comprising an anti-transferrin receptor antibody or antigen binding fragment thereof conjugated to an ASO sequence that hybridizes to a target sequence in exons 1-13 of the human DMPK mRNA excluding the CUG repeats and mediates RNA interference against the human DMPK mRNA preferentially in a muscle cell in a subject wherein the ASO sequence is 8 to 30 nucleotides in length.
2. The ASO conjugate of claim 1 , wherein the ASO sequence comprises at least one 2′ modified nucleotide, at least one modified internucleotide linkage, or at least one inverted abasic moiety.
3. The ASO conjugate of claim 1 , wherein mediation of RNA interference against the human DMPK mRNA modulates muscle atrophy or myotonic dystrophy in the subject.
4. The ASO conjugate of claim 1 , wherein the ASO sequence mediates RNA interference against the human DMPK mRNA via RNase H activity in the muscle cell.
5. The ASO conjugate of claim 1 , wherein the anti-transferrin receptor antibody or antigen binding fragment thereof binds to a transferrin receptor on the cell surface of the muscle cell.
6. The ASO conjugate of claim 1 , wherein the ASO sequence hybridizes to at least 8 contiguous nucleotides of the target sequence in the exons 1-13 of the human DMPK mRNA.
7. The ASO conjugate of claim 1 , wherein the ASO conjugate comprises a linker connecting the anti-transferrin receptor antibody or antigen binding fragment thereof to the ASO sequence.
8. The ASO conjugate of claim 2 , wherein the at least one 2′ modified nucleotide:
comprises 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl, 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′ DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O-NMA) modified nucleotide;
comprises locked nucleic acid (LNA) or ethylene nucleic acid (ENA); or
comprises a combination thereof.
9. The ASO conjugate of claim 2 , wherein the at least one modified internucleotide linkage comprises a phosphorothioate linkage or a phosphorodithioate linkage.
10. The ASO conjugate of claim 2 , wherein the ASO sequence comprises 3 or more 2′ modified nucleotides selected from 2′-O-methyl modified nucleotide and 2′-deoxy-2′-fluoro modified nucleotide.
11. The ASO conjugate of claim 1 , wherein the ASO conjugate has a drug to antibody ratio of from about 1 to about 4.
12. The ASO conjugate of claim 1 , wherein the ASO sequence comprises a 5′-terminal vinylphosphonate modified nucleotide.
13. The ASO conjugate of claim 1 , wherein the anti-transferrin receptor antibody or antigen-binding fragment thereof comprises a humanized antibody or antigen-binding fragment thereof, chimeric antibody or antigen-binding fragment thereof, monoclonal antibody or antigen-binding fragment thereof, monovalent Fab′, divalent Fab2, single-chain variable fragment (scFv), diabody, minibody, nanobody, single-domain antibody (sdAb), or camelid antibody or antigen-binding fragment thereof.
14. The ASO conjugate of claim 3 , wherein the muscle atrophy is associated with myotonic dystrophy type 1 (DM1).
15. The ASO conjugate of claim 3 , wherein the myotonic dystrophy is DM1.
16. The ASO conjugate of claim 1 , wherein the ASO conjugate is formulated for parenteral administration.
17. The ASO conjugate of claim 1 , wherein the ASO sequence that hybridizes to the target sequence of the human DMPK mRNA is selected from a group consisting of SEQ ID NOs: 6111-8814.
18. The ASO conjugate of claim 1 , wherein the ASO sequence that hybridizes to the target sequence of the human DMPK mRNA is selected from a group consisting of SEQ ID NOs: 11519-14222.
19. The ASO conjugate of claim 1 , wherein the ASO sequence that hybridizes to the target sequence in exons 2-13 of the human DMPK mRNA is selected from SEQ ID NOs: 11903, 11961, 11962, 11963, 12051, 12052, 12053, 12057, 12058, 12059, 12061, 12062, 12094, 12095, 12099, 12101, 12102, 12208, 12234, 12235, 12303, 12304, 12307, 12544, 12545, 12546, 12547, 12555, 12557, 12559, 12561, 12562, 12565, 12589, 12591, 12780, 12781, 12782, 12783, 12785, 12786, 12787, 12792, 12794, 12801, 12815, 12860, 12861, 12862, 12864, 13343, 13404, 13408, 13416, 13463, 13478, 13644, 13645, 13667, 13668, 13786, 13790, 14046, 14047, 14048, 14049, 14050, 14072, 14076, 14118, 14146, 14147, 14149, 14154, 14157, 14193, 14194, 14197, 14198, 14199, and 14200.
20. The ASO conjugate of claim 19 , wherein the ASO sequence that hybridizes to the target sequence in exons 4, 5, and 6 of the human DMPK mRNA is selected from SEQ ID NOs: 12102, 12234, 12303, 12304, and 12307.
21. The ASO conjugate of claim 19 , wherein the ASO sequence that hybridizes to the target sequence in exons 7 and 8 of the human DMPK mRNA is selected from SEQ ID NOs: 12544, 12545, 12546, 12547, 12555, 12557, 12559, 12561, 12562, 12565, 12589, 12783 and 12815.
22. The ASO conjugate of claim 19 , wherein the ASO sequence that hybridizes to the target sequence in exon 9 of the human DMPK mRNA is selected from SEQ ID NOs: 12860, 12861, 12862, and 12864.
23. The ASO conjugate of claim 19 , wherein the ASO sequence that hybridizes to the target sequence in exon 13 of the human DMPK mRNA is selected from SEQ ID NOs: 13343, 13463, 13478, 14047, 14048, 14049, 14072, 14146, 14147, 14199, and 14200.