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 small interfering RNA (siRNA)-antibody conjugate comprising an anti-transferrin receptor antibody or antigen-binding fragment thereof conjugated to an siRNA that hybridizes to a target sequence of human Murf1 mRNA and mediates RNA interference against the human Murf1 mRNA preferentially in a muscle cell, wherein the siRNA is a double stranded RNA comprising a guide strand and a passenger strand, and wherein the guide strand is a sequence selected from SEQ ID NOs: 651-702.
2. The siRNA-antibody conjugate of claim 1 , wherein the siRNA comprises at least one 2′ modified nucleotide, at least one modified internucleotide linkage, or at least one inverted abasic moiety.
3. The siRNA-antibody conjugate of claim 1 , wherein mediation of RNA interference against the human Murf1 mRNA in the muscle cell modulates muscle atrophy or myotonic dystrophy in a subject.
4. The siRNA-antibody conjugate of claim 1 , wherein the anti-transferrin receptor antibody or antigen-binding fragment thereof binds to a transferrin receptor on cell surface of the muscle cell.
5. The siRNA-antibody conjugate of claim 1 , wherein the muscle cell is a skeletal muscle cell or a cardiac muscle cell.
6. The siRNA-antibody conjugate of claim 1 , wherein the guide strand hybridizes to at least 8 contiguous bases of the target sequence of the human Murf1 mRNA.
7. The siRNA-antibody conjugate of claim 1 , wherein the siRNA is from about 19 to about 30 nucleotides in length.
8. The siRNA-antibody conjugate of claim 1 , wherein the siRNA antibody conjugate comprises a linker connecting the anti-transferrin receptor antibody or antigen-binding fragment thereof to the siRNA.
9. The siRNA-antibody conjugate of claim 3 , 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′-O-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.
10. The siRNA-antibody conjugate of claim 2 , wherein the at least one modified internucleotide linkage comprises a phosphorothioate linkage or a phosphorodithioate linkage.
11. The siRNA-antibody conjugate of claim 2 , wherein the siRNA comprises 3 or more 2′ modified nucleotides selected from 2′-O-methyl and 2′-deoxy-2′-fluoro.
12. The siRNA-antibody conjugate of claim 1 , wherein the siRNA-antibody conjugate has an siRNA to antibody ratio of from about 1 to about 4.
13. The siRNA-antibody conjugate of claim 1 , wherein the siRNA comprises a 5′-terminal vinylphosphonate modified nucleotide.
14. The siRNA-antibody conjugate of claim 4 , wherein the muscle atrophy is associated with myotonic dystrophy.
15. The siRNA-antibody conjugate of claim 1 , wherein the siRNA-antibody conjugate is formulated for parenteral administration.
16. The siRNA antibody conjugate of claim 1 , wherein the passenger strand is a sequence selected from SEQ ID NOs: 540-591.