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 the amino acid sequence of SEQ ID NO: 76; and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 75.
2 . The complex of claim 1 , wherein the VH of the antibody comprises an N-terminal pyroglutamate.
3 . The complex of claim 1 , wherein the complex is configured to promote transferrin receptor mediated internalization of the molecular payload into a muscle cell.
4 . The complex of claim 1 , wherein the molecular payload comprises an oligonucleotide.
5 . The complex of claim 4 , wherein the oligonucleotide comprises a region of complementarity to the muscle disease gene.
6 . The complex of claim 5 , wherein the region of complementarity is 20-30 nucleotides in length.
7 . The complex of claim 5 , wherein the oligonucleotide is 20-30 nucleotides in length.
8 . The complex of claim 7 , wherein the region of complementarity is 25-30 nucleotides in length.
9 . The complex of claim 4 , wherein the oligonucleotide is 15-35 nucleotides in length.
10 . The complex of claim 4 , wherein the oligonucleotide is 20-30 nucleotides in length.
11 . The complex of claim 4 , wherein the oligonucleotide comprises one or more modified nucleosides.
12 . The complex of claim 11 , wherein the one or more modified nucleosides comprise 2′-modified nucleosides.
13 . The complex of claim 12 , wherein each 2′-modified nucleoside is selected from the group consisting of: 2′-O-methyl, 2′-fluoro (2′-F), 2′-O-methoxyethyl (2′-MOE), and 2′,4′-bridged nucleosides.
14 . The complex of claim 11 , wherein the one or more modified nucleosides comprise 2′,4′-bridged nucleosides.
15 . The complex of claim 4 , wherein the oligonucleotide is a phosphorodiamidate morpholino oligomer.
16 . The complex of claim 4 , wherein the oligonucleotide comprises at least one modified internucleoside linkage.
17 . The complex of claim 4 , wherein the oligonucleotide comprises at least one phosphorothioate internucleoside linkage.
18 . The complex of claim 4 , 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.
19 . The complex of claim 4 , wherein the oligonucleotide is a mixmer oligonucleotide.
20 . The complex of claim 4 , wherein the oligonucleotide is an RNAi oligonucleotide that promotes RNAi-mediated cleavage of a mRNA transcript encoded by the muscle disease gene.
21 . The complex of claim 4 , wherein the oligonucleotide comprises a guide sequence for a genome editing nuclease.
22 . The complex of claim 1 , wherein the molecular payload comprises a peptide or polypeptide.
23 . The complex of claim 1 , wherein the molecular payload comprises a protein or fragment thereof.
24 . The complex of claim 23 , wherein the protein is an enzyme.
25 . The complex of claim 1 , wherein the molecular payload comprises a nucleic acid.
26 . The complex of claim 1 , wherein the molecular payload comprises a small molecule.
27 . The complex of claim 1 , wherein the muscle disease gene is GAA, GYS1, DNM2, DMD, DUX4, MYBPC3, MYH6, MYH7, TNNI3, TNNT2, ACVR1, FXN, GNE, MYH7, BAG3, CRYAB, DES, DNAJB6, FHL1, FLNC, LDB3, MYOT, PLEC, TTN, CLCN1, DMPK, CNBP, MTM1, PABPN1, or SCN4A.
28 . The complex of claim 1 , wherein the antibody is covalently linked to the molecular payload via a cleavable linker comprising a valine-citrulline sequence.
29 . The complex of claim 28 , comprising a structure of:
wherein n is 3 and m is 4, and wherein L1 comprises a spacer that is a substituted or unsubstituted aliphatic, substituted or unsubstituted heteroaliphatic, substituted or unsubstituted carbocyclylene, substituted or unsubstituted heterocyclylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, —O—, —N(R A )—, —S—, —C(═O)—, —C(═O)O—, —C(═O)NR A —, —NR A C(═O)—, —NR A C(═O)R A —, —C(═O)R A —, —NR A C(═O)O—, —NR A C(═O)N(R A )—, —OC(═O)—, —OC(═O)O—, —OC(═O)N(R A )—, —S(O) 2 NR A —, —NR A S(O) 2 —, or a combination thereof, wherein each R A is independently hydrogen or substituted or unsubstituted alkyl.
30 . A composition comprising complexes, wherein each complex comprises an anti-transferrin receptor (TfR) antibody covalently linked to at least one molecular payload configured to modulate expression or activity of a muscle disease gene, wherein the antibody comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 76 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 75, wherein each anti-TfR antibody of the complexes is on average covalently linked to 1 to 3 molecular payloads.
31 . The composition of claim 30 , wherein the VH of the antibody comprises an N-terminal pyroglutamate.
32 . A method of modulating expression or activity of a muscle disease gene in muscle cells of a subject, the method comprising administering to the subject a composition comprising the complex of claim 1 .
33 . The method of claim 32 , wherein the subject is human.
34 . The method of claim 32 , wherein the subject is a cynomolgus.
35 . The method of claim 32 , wherein the subject expresses a disease allele of the muscle disease gene.
36 . The method of claim 32 , wherein the composition is intravenously administered to the subject.
37 . The method of claim 32 , wherein the subject has 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, Thomsen Disease, Myotonic Dystrophy Type I, Myotonic Dystrophy Type II, Myotubular Myopathy, Oculopharyngeal Muscular Dystrophy, or Paramyotonia Congenita.
38 . The method of claim 32 , wherein the VH of the antibody comprises an N-terminal pyroglutamate.
39 . A method of treating a muscle disease in a subject, the method comprising administering to the subject a composition comprising the complex of claim 1 .
40 . The method of claim 39 , wherein the subject is human.
41 . The method of claim 39 , wherein the composition is intravenously administered to the subject.
42 . The method of claim 39 , 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, Thomsen Disease, Myotonic Dystrophy Type I, Myotonic Dystrophy Type II, Myotubular Myopathy, Oculopharyngeal Muscular Dystrophy, and Paramyotonia Congenita.