Muscle targeting complexes and uses thereof for treating myotonic 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 a DMPK allele comprising a disease-associated-repeat. In some embodiments, the molecular payload is an oligonucleotide, such as an antisense oligonucleotide or RNAi oligonucleotide.
1. A composition comprising complexes, wherein each complex comprises an anti-transferrin receptor (TfR) antibody covalently linked to at least one oligonucleotides, wherein the antibody is a Fab and comprises 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, wherein each anti-TfR antibody of the complexes is on average covalently linked to 1 to 3 oligonucleotides, and wherein the oligonucleotide targets a Dystrophia Myotonica Protein Kinase (DMPK) RNA and comprises a region of complementarity to a sequence as set forth in any one of SEQ ID NO: 384-619, wherein the region of complementarity is 15-25 nucleotides in length.
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 oligonucleotide comprises a 5′-X—Y—Z-3′ formula, wherein X and Z are flanking regions comprising one or more 2′-modified nucleosides selected from the group consisting of: 2′-O-methyl, 2′-fluoro, 2′-O-methoxyethyl, 2′,4′-bridged nucleosides, and a combination thereof, and wherein Y is a gap region and each nucleoside in Y is a 2′-deoxyribonucleoside.
4. The composition of claim 1 , wherein the oligonucleotide is 15-30 nucleotides in length.
5. The composition of claim 1 , wherein the oligonucleotide comprises at least 15 consecutive nucleotides of a sequence set forth in any one of SEQ ID NOs: 148-383, wherein any one or more of the thymine bases (T's) in the oligonucleotide is optionally a uracil base (U).
6. The composition of claim 1 , wherein the oligonucleotide comprises one or more phosphorothioate internucleoside linkages.
7. The composition of claim 1 , wherein the antibody is covalently linked to each oligonucleotide via a linker.
8. The composition of claim 7 , wherein the linker comprises a cleavable linker.
9. The composition of claim 8 , wherein the linker comprises a valine-citrulline sequence.
10. The composition of claim 9 , wherein each complex comprises a structure of:
wherein n is 3 and m is 4, 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.
11. The composition of claim 10 , wherein L1 comprises
12. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 395.
13. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 398.
14. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 408.
15. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 410.
16. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 416.
17. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 418.
18. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 424.
19. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 426.
20. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 431.
21. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 432.
22. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 437.
23. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 439.
24. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 448.
25. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 451.
26. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 454.
27. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 458.
28. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 484.
29. The composition of claim 1 , wherein the region of complementarity is to a sequence as set forth in SEQ ID NO: 500.
30. The composition of claim 1 , wherein the oligonucleotide comprises at least 15 consecutive nucleotides of a sequence set forth in any one of SEQ ID NOs: 159, 162, 172, 174, 180, 182, 188, 190, 195, 196, 201, 203, 212, 215, 218, 222, 248, and 264, wherein any one or more of the thymine bases (T's) in the oligonucleotide is optionally a uracil base (U).