MUSCLE TARGETING COMPLEXES AND USES THEREOF FOR TREATING POMPE DISEASE

1 . A complex comprising a muscle-targeting agent covalently linked to an oligonucleotide comprising an antisense strand targeting a glycogen synthase 1 (GYS1) RNA, wherein the antisense strand comprises a region of complementarity to a target sequence as set forth in any one of SEQ ID NOs: 266-7493 and SEQ ID NOs: 162-186, wherein the region of complementarity is at least 12 consecutive nucleosides in length.

2 . The complex of claim 1 , wherein the muscle-targeting agent is an anti-transferrin receptor 1 (TfR1) antibody.

3 . The complex of claim 1 , wherein the region of complementary is at least 16 nucleotides in length.

4 . The complex of claim 1 , wherein the antisense strand comprises the nucleotide sequence of any one of SEQ ID NOs: 7494-11107 and 212-236, wherein each of the Us are optionally and independently Ts.

5 . The complex of claim 1 , wherein the antisense strand is 18-25 nucleotides in length, optionally wherein the antisense strand is 23 nucleotides in length.

6 . The complex of claim 1 , wherein the oligonucleotide further comprises a sense strand which comprises at least 18 consecutive nucleosides complementary to the antisense strand.

7 . The complex of claim 6 , wherein the antisense strand comprises the nucleotide sequence of any one of SEQ ID NOs: 212-236 and the sense strand comprises the nucleotide sequence of any one of SEQ ID NOs: 187-211, wherein each of the Us are optionally and independently Ts.

8 . The complex of claim 1 , wherein the oligonucleotide comprises one or more modified nucleosides selected from: 2′-fluoro (2′-F), 2′-O-methyl (2′-O-Me), 2′-O-methoxyethyl (2′-MOE), 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), 2′-O—N-methylacetamido (2′-O-NMA)).

9 . (canceled)

10 . The complex of claim 8 , wherein one or more modified nucleosides are 2′ modified nucleotides, optionally wherein each 2′ modified nucleotide is 2′-O-methyl or 2′-fluoro (2′-F).

11 . The complex of claim 1 , wherein the oligonucleotide comprises one or more phosphorothioate internucleoside linkages.

12 . (canceled)

13 . The complex of claim 1 , wherein the antisense strand comprises two phosphorothioate internucleoside linkages.

14 . The complex of claim 3 , wherein the two internucleoside linkages at the 3′ end of the antisense strand are phosphorothioate internucleoside linkages.

15 . The complex of claim 1 , wherein the antisense strand is selected from the modified version of SEQ ID NOs: 212-236 listed in Table 8.

16 . The complex of claim 6 , wherein the sense strand is selected from the modified version of SEQ ID NOs: 187-211 listed in Table 8.

17 . The complex of claim 1 , wherein the oligonucleotide is an siRNA molecule selected from the siRNAs listed in Table 8.

18 .- 20 . (canceled)

21 . The complex of claim 1 , wherein the muscle targeting agent and the oligonucleotide are covalently linked via a linker, optionally wherein the linker comprises a valine-citrulline sequence.

22 . A method of reducing GYS1 expression in a muscle cell, the method comprising contacting the muscle cell with an effective amount of the complex of claim 1 for promoting internalization of the RNAi oligonucleotide to the muscle cell.

23 . (canceled)

24 . A method of treating Pompe disease (PD), the method comprising administering to a subject in need thereof an effective amount of the complex of claim 1 .

25 . An siRNA oligonucleotide comprising a pair of an antisense strand and a sense strand selected from the following pairs:

Antisense strand:

(SEQ ID NO: 212)

5′-fUfCmGfAmAfUmUfCmAfUmCfCmUfCmCfCmAfGmUfCmC

*fU*mC-3′

Sense strand:

(SEQ ID NO: 187)

5′-mGmGfAmCfUmGfGmGfAmGfGmAfUmGfAmAfUmUfCmGfA

-3′

Antisense strand:

(SEQ ID NO: 213)

5′-fGfUmCfGmAfAmUfUmCfAmUfCmCfUmCfCmCfAmGfUmC

*fC*mU-3′

Sense strand:

(SEQ ID NO: 188)

5′-mGmAfCmUfGmGfGmAfGmGfAmUfGmAfAmUfUmCfGmAfC

-3′

Antisense strand:

(SEQ ID NO: 214)

5′-fUfAmGfAmUfGmCfCmAfCmCfCmAfCmCfUmUfGmUfUmA

*fG*mC-3′

Sense strand:

(SEQ ID NO: 189)

5′-mUmAfAmCfAmAfGmGfUmGfGmGfUmGfGmCfAmUfCmUfA

-3′

Antisense strand:

(SEQ ID NO: 215)

5′-fUfGmUfAmGfAmUfGmCfCmAfCmCfCmAfCmCfUmUfGmU

*fU*mA-3′

Sense strand:

(SEQ ID NO: 190)

5′-mAmCfAmAfGmGfUmGfGmGfUmGfGmCfAmUfCmUfAmCfA

-3′

Antisense strand:

(SEQ ID NO: 216)

5′-fCfUmUfCmGfCmCfUmUfCmGfUmCfUmGfCmAfGmCfAmC

*fC*mG-3′

Sense strand:

(SEQ ID NO: 191)

5′-mGmUfGmCfUmGfCmAfGmAfCmGfAmAfGmGfCmGfAmAfG

-3′

Antisense strand:

(SEQ ID NO: 217)

5′-fAfCmCfUmUfCmGfCmCfUmUfCmGfUmCfUmGfCmAfGmC

*fA*mC-3′

Sense strand:

(SEQ ID NO: 192)

5′-mGmCfUmGfCmAfGmAfCmGfAmAfGmGfCmGfAmAfGmGfU

-3′

Antisense strand:

(SEQ ID NO: 218)

5′-fUfCmAfCmCfUmUfCmGfCmCfUmUfCmGfUmCfUmGfCmA

*fG*mC-3′

Sense strand:

(SEQ ID NO: 193)

5′-mUmGfCmAfGmAfCmGfAmAfGmGfCmGfAmAfGmGfUmGfA

-3′

Antisense strand:

(SEQ ID NO: 219)

5′-fGfUmCfAmCfCmUfUmCfGmCfCmUfUmCfGmUfCmUfGmC

*fA*mG-3′

Sense strand:

(SEQ ID NO: 194)

5′-mGmCfAmGfAmCfGmAfAmGfGmCfGmAfAmGfGmUfGmAfC

-3′

Antisense strand:

(SEQ ID NO: 220)

5′-fCfCmCfUmUfGmCfUmGfUmUfCmAfUmGfGmAfAmUfCmC

*fA*mG-3′

Sense strand:

(SEQ ID NO: 195)

5′-mGmGfAmUfUmCfCmAfUmGfAmAfCmAfGmCfAmAfGmGfG

-3′

Antisense strand:

(SEQ ID NO: 221)

5′-fCfCmAfGmGfUmUfGmUfUmGfUmAfGmAfAmGfUmCfCmA

*fC*mG-3′

Sense strand:

(SEQ ID NO: 196)

5′-mUmGfGmAfCmUfUmCfUmAfCmAfAmCfAmAfCmCfUmGfG

-3′

Antisense strand:

(SEQ ID NO: 222)

5′-fCfUmGfAmGfCmAfGmAfUmAfGmUfUmGfAmGfCmCfGmA

*fG*mC-3′

Sense strand:

(SEQ ID NO: 197)

5′-mUmCfGmGfCmUfCmAfAmCfUmAfUmCfUmGfCmUfCmAfG

-3′

Antisense strand:

(SEQ ID NO: 223)

5′-fUfCmUfGmAfGmCfAmGfAmUfAmGfUmUfGmAfGmCfCmG

*fA*mG-3′

Sense strand:

(SEQ ID NO: 198)

5′-mCmGfGmCfUmCfAmAfCmUfAmUfCmUfGmCfUmCfAmGfA

-3′

Antisense strand:

(SEQ ID NO: 224)

5′-fCfAmCfUmCfUmGfAmGfCmAfGmAfUmAfGmUfUmGfAmG

*fC*mC-3′

Sense strand:

(SEQ ID NO: 199)

5′-mCmUfCmAfAmCfUmAfUmCfUmGfCmUfCmAfGmAfGmUfG

-3′

Antisense strand:

(SEQ ID NO: 225)

5′-fUfUmCfAmCfUmCfUmGfAmGfCmAfGmAfUmAfGmUfUmG

*fA*mG-3′

Sense strand:

(SEQ ID NO: 200)

5′-mCmAfAmCfUmAfUmCfUmGfCmUfCmAfGmAfGmUfGmAfA

-3′

Antisense strand:

(SEQ ID NO: 226)

5′-fCfCmAfGmCfAmUfCmUfUmGfUmUfCmAfUmGfUmCfGmG

*fG*mA-3′

Sense strand:

(SEQ ID NO: 201)

5′-mCmCfGmAfCmAfUmGfAmAfCmAfAmGfAmUfGmCfUmGfG

-3′

Antisense strand:

(SEQ ID NO: 227)

5′-fUfGmGfCmUfCmUfCmUfUmCfAmUfCmAfUmAfGmUfGmA

*fA*mG-3′

Sense strand:

(SEQ ID NO: 202)

5′-mUmCfAmCfUmAfUmGfAmUfGmAfAmGfAmGfAmGfCmCfA

-3′

Antisense strand:

(SEQ ID NO: 228)

5′-fAfUmUfCmGfGmCfGmGfAmUfGmGfUmGfGmUfCmAfGmG

*fA*mU-3′

Sense strand:

(SEQ ID NO: 203)

5′-mCmCfUmGfAmCfCmAfCmCfAmUfCmCfGmCfCmGfAmAfU

-3′

Antisense strand:

(SEQ ID NO: 229)

5′-fCfUmGfCmGfAmUfGmUfGmUfUmCfCmUfCmCfAmUfGmA

*fA*mG-3′

Sense strand:

(SEQ ID NO: 204)

5′-mUmCfAmUfGmGfAmGfGmAfAmCfAmCfAmUfCmGfCmAfG

-3′

Antisense strand:

(SEQ ID NO: 230)

5′-fUfCmUfGmCfGmAfUmGfUmGfUmUfCmCfUmCfCmAfUmG

*fA*mA-3′

Sense strand:

(SEQ ID NO: 205)

5′-mCmAfUmGfGmAfGmGfAmAfCmAfCmAfUmCfGmCfAmGfA

-3′

Antisense strand:

(SEQ ID NO: 231)

5′-fGfAmAfUmCfAmUfCmCfAmGfGmCfUmGfCmGfGmAfAmC

*fC*mG-3′

Sense strand:

(SEQ ID NO: 206)

5′-mGmUfUmCfCmGfCmAfGmCfCmUfGmGfAmUfGmAfUmUfC

-3′

Antisense strand:

(SEQ ID NO: 232)

5′-fGfCmCfAmUfGmUfGmGfCmGfCmGfCmAfGmAfCmAfUmA

*fU*mA-3′

Sense strand:

(SEQ ID NO: 207)

5′-mUmAfUmGfUmCfUmGfCmGfCmGfCmCfAmCfAmUfGmGfC

-3′

Antisense strand:

(SEQ ID NO: 233)

5′-fGfCmUfCmUfCmUfUmCfAmUfCmAfUmAfGmUfGmAfAmG

*fU*mC-3′

Sense strand:

(SEQ ID NO: 208)

5′-mCmUfUmCfAmCfUmAfUmGfAmUfGmAfAmGfAmGfAmGfC

-3′

Antisense strand:

(SEQ ID NO: 234)

5′-fAfAmCfUmUfCmUfUmCfAmCfAmUfUmCfAmGfCmCfCmA

*fU*mU-3′

Sense strand:

(SEQ ID NO: 209)

5′-mUmGfGmGfCmUfGmAfAmUfGmUfGmAfAmGfAmAfGmUfU

-3′

Antisense strand:

(SEQ ID NO: 235)

5′-fUfUmGfCmUfGmUfUmCfAmUfGmGfAmAfUmCfCmAfGmU

*fG*mU-3′

Sense strand:

(SEQ ID NO: 210)

5′-mAmCfUmGfGmAfUmUfCmCfAmUfGmAfAmCfAmGfCmAfA

-3′

Antisense strand:

(SEQ ID NO: 236)

5′-fUfUmCfUmCfCmAfGmGfUmUfGmUfUmGfUmAfGmAfAmG

*fU*mC-3′

Sense strand:

(SEQ ID NO: 211)

5′-mCmUfUmCfUmAfCmAfAmCfAmAfCmCfUmGfGmAfGmAfA

-3′

wherein “m” is 2′-O-methyl (2′-O-Me); “f” is 2′-fluoro (2′-F); “*” indicates phosphorothioate internucleoside linkage; and the absence of “*” between nucleosides indicate a phosphodiester linkage.

26 . A composition comprising the siRNA oligonucleotide of claim 25 in sodium salt form.