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 Atrogin-1 mRNA and mediates RNA interference against the human Atrogin-1 mRNA preferentially in a muscle cell, wherein the siRNA comprises a guide strand and a passenger strand, and the guide strand comprises a sequence selected from SEQ ID NO: 318, SEQ ID NO: 321, SEQ ID NO: 325, SEQ ID NO: 327, SEQ ID NO:333, SEQ ID NO: 337, SEQ ID NO:339, SEQ ID NO:342, SEQ ID NO:344, SEQ ID NO: 346, SEQ ID NO:348, SEQ ID NO:350, SEQ ID NO:353, SEQ ID NO:358, SEQ ID NO:359, SEQ ID NO:361, SEQ ID NO:363, SEQ ID NO:365, SEQ ID NO:366 and SEQ ID NO: 368.
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 the anti-transferrin receptor antibody or antigen-binding fragment thereof binds to a transferrin receptor on the cell surface of the muscle cell.
4. The siRNA-antibody conjugate of claim 1 , wherein the muscle cell is a skeletal muscle cell or a cardiac muscle cell.
5. The siRNA-antibody conjugate of claim 1 , wherein the guide strand or the passenger strand is from about 19 to about 30 nucleotides in length.
6. 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.
7. The siRNA-antibody 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′-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.
8. The siRNA-antibody conjugate of claim 2 , wherein the at least one modified internucleotide linkage comprises a phosphorothioate linkage or a phosphorodithioate linkage.
9. 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.
10. The siRNA-antibody conjugate of claim 1 , wherein the siRNA-antibody conjugate has a drug/siRNA to antibody ratio of from about 1 to 11 about 4.
11. The siRNA-antibody conjugate of claim 1 , wherein the siRNA comprises a 5′-terminal vinylphosphonate modified nucleotide.
12. The siRNA-antibody 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.
13. The siRNA-antibody conjugate of claim 1 , wherein the siRNA-antibody conjugate is formulated for parenteral, oral, intranasal, buccal, rectal, or transdermal administration.