Muscle targeting complexes and uses thereof for treating facioscapulohumeral muscular dystrophy
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 DUX4. In some embodiments, the molecular payload is an oligonucleotide, such as an antisense oligonucleotide or RNAi oligonucleotide.
1. A composition comprising complexes comprising an anti-transferrin receptor (TfR) antibody covalently linked to at least one oligonucleotide, wherein the antibody is a Fab comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 101 and a light chain comprising the amino acid sequence of SEQ ID NO: 90, and wherein each anti-TfR antibody of the complexes is on average covalently linked to 1 to 3 oligonucleotides.
2. The composition of claim 1 , wherein the heavy chain of the antibody comprises an N-terminal pyroglutamate.
3. The composition of claim 1 , wherein the equilibrium dissociation constant (KD) of binding of the antibody to the transferrin receptor is in a range from 10 −11 M to 10 −6 M.
4. The composition of claim 1 , wherein the oligonucleotide is single stranded.
5. The composition of claim 1 , wherein the oligonucleotide is double stranded.
6. The composition of claim 1 , wherein the oligonucleotide is an siRNA.
7. The composition of claim 1 , wherein the oligonucleotide comprises one or more modified nucleosides.
8. The composition of claim 7 , wherein the one or more modified nucleosides comprise 2′-modified nucleosides.
9. The composition of claim 1 , wherein the oligonucleotide comprises one or more modified internucleoside linkages.
10. The composition of claim 9 , wherein the oligonucleotide comprises one or more phosphorothioate internucleoside linkages.
11. The composition of claim 1 , wherein the antibody is covalently linked to each oligonucleotide via a linker.
12. The composition of claim 11 , wherein the linker comprises a cleavable linker.
13. The composition of claim 12 , wherein the linker comprises a valine-citrulline sequence.
14. The composition of claim 1 , wherein each complex comprises 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.
15. A method of delivering an oligonucleotide to a subject, comprising administering to the subject the composition of claim 8 .
16. The method of claim 15 , wherein the oligonucleotide is delivered to a muscle cell of the subject.
17. The method of claim 15 , wherein the oligonucleotide is single stranded.
18. The method of claim 15 , wherein the oligonucleotide is double stranded.
19. The method of claim 15 , wherein the oligonucleotide is an siRNA.
20. The method of claim 15 , wherein the subject is human.
21. The method of claim 15 , wherein the subject has a muscle disorder.
22. The method of claim 21 , wherein the muscle disorder is a muscular dystrophy.
23. The method of claim 22 , wherein the muscular dystrophy is DMD.
24. The method of claim 22 , wherein the muscular dystrophy is DM1.
25. The method of claim 22 , wherein the muscular dystrophy is FSHD.