MUSCLE TARGETING COMPLEXES AND USES THEREOF FOR TREATING HYPERTROPHIC CARDIOMYOPATHY
Aspects of the disclosure relate to complexes comprising a muscle-targeting agent covalently linked to a molecular payload. In some embodiments, the muscle-targeting agent specifically binds to an internalizing cell surface receptor on muscle cells. In some embodiments, the molecular payload inhibits expression or activity of a MYH7 allele comprising a disease-associated mutation, e.g., for the treatment of hypertrophic cardiomyopathy. In some embodiments, the molecular payload is an oligonucleotide, such as an antisense oligonucleotide or RNAi oligonucleotide.
1 .- 71 . (canceled)
72 . A complex comprising an anti-transferrin receptor antibody covalently linked to an oligonucleotide that targets an MYH7 RNA,
wherein the oligonucleotide is 15-35 nucleotides in length and comprises a region of complementarity to the MYH7 RNA sequence as set forth in SEQ ID NO: 15, wherein the region of complementarity is at least 12 nucleotides in length; and
wherein the anti-transferrin receptor antibody binds in the range of C89 to F760 of human transferrin receptor protein 1 (TfR1) having an amino acid sequence as set forth in SEQ ID NO: 1.
73 . The complex of claim 72 , wherein the anti-transferrin receptor antibody is in the form of a ScFv, Fab fragment, Fab′ fragment, F(ab′)2 fragment, or Fv fragment.
74 . The complex of claim 72 , wherein the anti-transferrin receptor antibody binds human TfR1 with a K D of 10 −11 M to 10 −6 M.
75 . The complex of claim 72 , wherein the anti-transferrin receptor antibody is a humanized antibody.
76 . The complex of claim 72 , wherein the oligonucleotide comprises one or more modified nucleosides.
77 . The complex of claim 76 , wherein the one or more modified nucleosides are 2′-modified nucleosides selected from the group consisting of: 2′-O-methyl, 2′-fluoro, 2′-O-methoxyethyl, and 2′, 4′-bridged nucleosides.
78 . The complex of claim 72 , wherein the oligonucleotide comprises one or more modified internucleoside linkages.
79 . The complex of claim 78 , wherein the one or more modified internucleoside linkage are phosphorothioate linkages.
80 . The complex of claim 72 , wherein the oligonucleotide comprises one or more phosphorodiamidate morpholinos.
81 . The complex of claim 72 , wherein the oligonucleotide is a phosphorodiamidate morpholino oligomer (PMO).
82 . The complex of claim 72 , wherein the oligonucleotide comprises a region of complementarity to at least 15 consecutive nucleotides of SEQ ID NO: 15.
83 . The complex of claim 72 , wherein the anti-transferrin receptor antibody is covalently linked to the oligonucleotide via a cleavable linker.
84 . The complex of claim 83 , wherein the cleavable linker comprises a valine-citrulline sequence.
85 . The complex of claim 72 , wherein the anti-transferrin receptor antibody is covalently linked to the oligonucleotide via conjugation to a lysine residue or a cysteine residue of the anti-transferrin receptor antibody.
86 . The complex of claim 72 , wherein the complex is configured to promote transferrin receptor mediated internalization of the oligonucleotide into a muscle cell.
87 . The complex of claim 72 , wherein the oligonucleotide brings about reduction of expression level of MYH7 in the muscle cell.
88 . The complex of claim 72 , wherein the oligonucleotide is an antisense oligonucleotide.
89 . The complex of claim 72 , wherein the oligonucleotide is an siRNA.
90 . A method of reducing MYH7 expression in a muscle cell, the method comprising contacting the muscle cell with a complex comprising an anti-transferrin receptor antibody covalently linked to an oligonucleotide that targets a MYH7 RNA,
wherein the oligonucleotide is 15 to 35 nucleotides in length and comprises a region of complementarity to the MYH7 RNA sequence as set forth in SEQ ID NO: 15, wherein the region of complementarity is at least 12 nucleotides in length; and
wherein the anti-transferrin receptor antibody binds in the range of C89 to F760 of human transferrin receptor protein 1 (TfR1) having an amino acid sequence as set forth in SEQ ID NO: 1.
91 . A method of treating hypertrophic cardiomyopathy (HCM) in a subject, the method comprising administering to the subject a complex comprising an anti-transferrin receptor antibody covalently linked to an oligonucleotide that targets a MYH7 RNA,
wherein the oligonucleotide is 15 to 35 nucleotides in length and comprises a region of complementarity to the MYH7 RNA sequence as set forth in SEQ ID NO: 15, wherein the region of complementarity is at least 12 nucleotides in length; and
wherein the anti-transferrin receptor antibody binds in the range of C89 to F760 of human transferrin receptor protein 1 (TfR1) having an amino acid sequence as set forth in SEQ ID NO: 1.