MUSCLE-TARGETING COMPLEXES AND USES THEREOF
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 activity of a disease allele associated with muscle disease. In some embodiments, the molecular payload is an oligonucleotide, such as an antisense oligonucleotide or RNAi oligonucleotide.
1 . A complex comprising an anti-transferrin receptor (TfR) antibody covalently linked to a molecular payload configured to modulate expression or activity of a muscle disease gene, wherein the antibody comprises:
a heavy chain variable region (VH) comprising an amino acid sequence at least 95% identical to SEQ ID NO: 77; and/or a light chain variable region (VL) comprising an amino acid sequence at least 95% identical to SEQ ID NO: 78.
2 . The complex of claim 1 , wherein the antibody comprises:
a VH comprising the amino acid sequence of SEQ ID NO: 77 and a VL comprising the amino acid sequence of SEQ ID NO: 78.
3 . The complex of claim 1 , wherein the antibody is selected from the group consisting of a Fab fragment, a Fab′ fragment, a F(ab′)2 fragment, a scFv, a Fv, and a full-length IgG.
4 . The complex of claim 3 , wherein the antibody is a Fab fragment.
5 . The complex of claim 4 , wherein the antibody comprises:
a heavy chain comprising an amino acid sequence at least 85% identical to SEQ ID NO: 102; and/or a light chain comprising an amino acid sequence at least 85% identical to SEQ ID NO: 93.
6 . The complex of claim 4 , wherein the antibody comprises:
a heavy chain comprising the amino acid sequence of SEQ ID NO: 102; and a light chain comprising the amino acid sequence of SEQ ID NO: 93.
7 . The complex of claim 1 , wherein the antibody does not specifically bind to the transferrin binding site of the transferrin receptor and/or wherein the antibody does not inhibit binding of transferrin to the transferrin receptor.
8 . The complex of claim 1 , wherein the antibody is cross-reactive with extracellular epitopes of two or more of a human, non-human primate and rodent transferrin receptor.
9 . The complex of claim 1 , wherein the complex is configured to promote transferrin receptor mediated internalization of the molecular payload into a muscle cell.
10 . The complex of claim 1 , wherein the molecular payload is an oligonucleotide.
11 . The complex of claim 10 , wherein the oligonucleotide comprises a region of complementarity to a muscle disease gene having a gain-of-function disease allele.
12 . The complex of claim 10 , wherein the oligonucleotide comprises at least one modified internucleoside linkage.
13 . (canceled)
14 . The complex of claim 10 , wherein the oligonucleotide comprises one or more modified nucleosides.
15 . (canceled)
16 . The complex of claim 10 , wherein the oligonucleotide is a gapmer oligonucleotide that directs RNAse H-mediated cleavage of an mRNA transcript encoded by the muscle disease gene in a cell.
17 . (canceled)
18 . The complex of claim 10 , wherein the oligonucleotide is an RNAi oligonucleotide that promotes RNAi-mediated cleavage of a mRNA transcript encoded by the muscle disease gene.
19 .- 21 . (canceled)
22 . The complex of claim 1 , wherein the antibody is covalently linked to the molecular payload via a cleavable linker, optionally wherein the cleavable linker comprises a valine-citrulline sequence.
23 . (canceled)
24 . The complex of claim 1 , wherein the antibody is covalently linked to the molecular payload via conjugation to a lysine residue or a cysteine residue of the antibody.
25 . The complex of claim 1 , wherein modulating expression or activity of a muscle disease gene comprises reducing expression of RNA and/or protein.
26 . A method of modulating expression or activity of a muscle disease gene in a cell, the method comprising contacting the cell with the complex of claim 1 in an amount effective for promoting internalization of the molecular payload to the cell, optionally wherein the cell is a muscle cell, optionally wherein the muscle disease is a disease selected from the group consisting of: Adult Pompe Disease, Centronuclear myopathy (CNM), Duchenne Muscular Dystrophy, Facioscapulohumeral Muscular Dystrophy (FSHD), Familial Hypertrophic Cardiomyopathy, Fibrodysplasia Ossificans Progressiva (FOP), Friedreich's Ataxia (FRDA), Inclusion Body Myopathy 2, Laing Distal Myopathy, Myofibrillar Myopathy, Myotonia Congenita (autosomal dominant form, Thomsen Disease), Myotonic Dystrophy Type I, Myotonic Dystrophy Type II, Myotubular Myopathy, Oculopharyngeal Muscular Dystrophy, and Paramyotonia Congenita.
27 . (canceled)
28 . A method of treating a subject having a muscle disease, the method comprising administering to the subject an effective amount of the complex of claim 1 optionally wherein the muscle disease is a disease selected from the group consisting of: Adult Pompe Disease, Centronuclear myopathy (CNM), Duchenne Muscular Dystrophy, Facioscapulohumeral Muscular Dystrophy (FSHD), Familial Hypertrophic Cardiomyopathy, Fibrodysplasia Ossificans Progressiva (FOP), Friedreich's Ataxia (FRDA), Inclusion Body Myopathy 2, Laing Distal Myopathy, Myofibrillar Myopathy, Myotonia Congenita (autosomal dominant form, Thomsen Disease), Myotonic Dystrophy Type I, Myotonic Dystrophy Type II, Myotubular Myopathy, Oculopharyngeal Muscular Dystrophy, and Paramyotonia Congenita.