RNAi Agents for Inhibiting Expression of DUX4, Compositions Thereof, And Methods of Use

1 . An RNAi agent for inhibiting expression of a double homeobox 4 (DUX4) gene, comprising:

i. an antisense strand comprising at least 17 contiguous nucleotides differing by 0 or 1 nucleotides from SEQ ID NO:164; and

ii. a sense strand comprising a nucleotide sequence that is at least partially complementary to the antisense strand.

2 . The RNAi agent of claim 1 , wherein the antisense strand comprises nucleotides 2-18 of SEQ ID NO:164.

3 - 4 . (canceled)

5 . The RNAi agent of claim 2 , wherein all or substantially all of the nucleotides are modified nucleotides.

6 . (canceled)

7 . The RNAi agent of claim 2 , wherein all or substantially all of the modified nucleotides are 2′-O-methyl nucleotides, 2′-fluoro nucleotides, or combinations thereof.

8 . The RNAi agent of claim 5 , wherein the antisense strand comprises the nucleotide sequence of SEQ ID NO:164.

9 . The RNAi agent of claim 8 , wherein the sense strand comprises the nucleotide sequence of SEQ ID NO: 183.

10 . (canceled)

11 . The RNAi agent of claim 5 , wherein the RNAi agent is linked to a targeting ligand.

12 . The RNAi agent of claim 11 , wherein the targeting ligand is linked to the sense strand.

13 . The RNAi agent of claim 12 , wherein the targeting ligand is linked to the 5′ terminal end of the sense strand.

14 . The RNAi agent of claim 13 , wherein the targeting ligand has affinity for a skeletal muscle cell and/or a cell receptor expressed on a skeletal muscle cell.

15 . The RNAi agent of claim 11 , wherein the targeting ligand is selected from the group consisting of:

Compound

Number

Formula

40b

41b

42b

43b

44b

45b

46b

47b

48b

49b

50b

51b

52b

53b

54b

55b

56b

57b

58b

59b

60b

ανβ6 Peptide 1

or a pharmaceutically acceptable salt thereof, wherein

indicates the point of connection to the RNAi agent.

16 . The RNAi agent of claim 5 , wherein the RNAi agent is linked to a pharmacokinetic/pharmacodynamic (PK/PD) modulator, preferably wherein the PK/PD modulator is linked to the sense strand.

17 . (canceled)

18 . The RNAi agent of claim 17 , wherein the PK/PD modulator is linked to the 3′ terminal end of the sense strand.

19 . The RNAi agent of claim 16 , wherein the PK/PD modulator is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof, wherein indicates the point of connection to the RNAi agent.

20 . The RNAi agent of claim 16 , wherein the PK/PD modulator is selected from the group consisting of:

wherein R Z comprises the RNAi agent.

21 .- 30 . (canceled)

31 . An RNAi agent of claim 1 , comprising:

(i) an antisense strand that consists of, consists essentially of, or comprises the modified nucleotide sequence (5′→3′) usAfsGfsAfauuucacGfgAfaGfaacasg (SEQ ID NO:82), and a sense strand that consists of, consists essentially of, or comprises the modified nucleotide sequence (5′→3′) cuguucuuCfCfGfugaaauucua (SEQ ID NO:149); or

(ii) an antisense strand that consists of, consists essentially of, or comprises the modified nucleotide sequence (5′→3′) cPrpusAfsGfsAfauuucacGfgAfaGfaacasg (SEQ ID NO:84), and a sense strand that consists of, consists essentially of, or comprises the modified nucleotide sequence (5′→3′) cuguucuuCfCfGfugaaauucua (SEQ ID NO:149); or

(iii) an antisense strand that consists of, consists essentially of, or comprises the modified nucleotide sequence (5′→3′) cPrpusAfsgsAfauuucacGfgAfaGfaacasg (SEQ ID NO:100), and a sense strand that consists of, consists essentially of, or comprises the modified nucleotide sequence (5′→3′) cuguucuuCfCfGfugaaauucua (SEQ ID NO:149); or

(iv) an antisense strand that consists of, consists essentially of, or comprises the modified nucleotide sequence (5′→3′) cPrpusAfsGfsaauuucacGfgAfaGfaacasg (SEQ ID NO:101), and a sense strand that consists of, consists essentially of, or comprises the modified nucleotide sequence (5′→3′) cuguucuuCfCfGfugaaauucua (SEQ ID NO:149);

wherein a represents 2′-O-methyl adenosine: c represents 2′-O-methyl cytidine: g represents 2′-O-methyl guanosine: u represents 2′-O-methyl uridine: Af represents 2′-fluoro adenosine: Cf represents 2′-fluoro cytidine: Gf represents 2′-fluoro guanosine; Uf represents 2′-fluoro uridine; cPrpu represents a 5′-cyclopropyl phosphonate-2′-O-methyl uridine; s represents a phosphorothioate linkage; and wherein the respective sense strand further optionally includes an inverted abasic residue at the 3′ terminal end of the nucleotide sequence and at the 5′ terminal end of the nucleotide sequence; and the sense strand also optionally includes a targeting ligand that is covalently linked to the inverted abasic residue at the 5′ terminal end of the sense strand, wherein the targeting ligand has affinity for skeletal muscle cells and/or a receptor present on skeletal muscle cells, and wherein the sense strand further optionally includes a PK/PD modulator that is covalently linked to the inverted abasic residue at the 3′ terminal end of the sense strand.

32 .- 37 . (canceled)

38 . The RNAi agent of claim 1 , wherein the RNAi agent is a pharmaceutically acceptable salt.

39 . (canceled)

40 . A pharmaceutical composition comprising the RNAi agent of claim 31 , wherein the composition further comprises a pharmaceutically acceptable excipient.

41 . A method for inhibiting expression of a DUX4 gene in a cell, the method comprising introducing into a cell an effective amount of the composition of claim 40 .

42 .- 45 . (canceled)

46 . A method of treating one or more symptoms or diseases that can be ameliorated at least in part by a reduction in DUX4 protein levels or a reduction in DUX4 mRNA levels, optionally wherein the disease is Facioscapulohumeral Muscular Dystrophy (FSHD), the method comprising administering to a human subject in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 40 .

47 .- 56 . (canceled)