Compositions and methods for inhibiting expression of TMPRSS6 gene

1. A double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of TMPRSS6 in a cell, wherein said dsRNA agent comprises a sense strand and an antisense strand forming a double-stranded region, wherein the antisense strand comprises the nucleotide sequence of any one of

5′-UCcAAAGGGcAGCUGAGCUdTsdT-3′ (SEQ ID NO: 577),

5′-UUCcAAAGGGcAGCUGAGCdTsdT-3′ (SEQ ID NO: 585),

5′-uAUUCcAAAGGGcAGCUGAdTsdT-3′ (SEQ ID NO: 593), or

5′-UuAUUCcAAAGGGcAGCUGdTsdT-3 (SEQ ID NO: 601),

wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U; dT is a deoxy-thymine nucleotide;

and s is a phosphorothioate linkage.

2. The dsRNA agent of claim 1 , wherein each strand is no more than 30 nucleotides in length.

3. The dsRNA agent of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide or at least 2 nucleotides.

4. The dsRNA agent of claim 1 , further comprising a ligand.

5. The dsRNA agent of claim 4 , wherein the ligand is conjugated to the 3′ end of the sense strand of the dsRNA agent.

6. A cell containing the dsRNA agent of claim 1 .

7. A pharmaceutical composition for inhibiting expression of a TMPRSS6 gene comprising the dsRNA agent of claim 1 .

8. The pharmaceutical composition of claim 7 , further comprising a lipid formulation.

9. A method of inhibiting TMPRSS6 expression in a cell, the method comprising:

(a) introducing into the cell the dsRNA agent of claim 1 ; and

(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a TMPRSS6 gene, thereby inhibiting expression of the TMPRSS6 gene in the cell.

10. The method of claim 9 , wherein the TMPRSS6 expression is inhibited by at least 30%.

11. A method of treating a disorder mediated by TMPRSS6 expression comprising administering to a human in need of such treatment a therapeutically effective amount of the dsRNA agent of claim 1 .

12. The method of claim 11 , wherein the human has a disorder associated with hemochromatosis, a β-thalassemia, or β-thalassemia intermedia.

13. The method of claim 11 , wherein the human has a β-thalassemia and wherein the administration of the dsRNA agent to the subject causes a decrease in iron in the serum of the subject by at least 10%.

14. The method of claim 11 , wherein the dsRNA agent is administered at a concentration of 0.01 mg/kg-5 mg/kg bodyweight of the subject.

15. The dsRNA agent of claim 1 , wherein the antisense strand consists of a nucleotide sequence selected from the group consisting of

5′-UCcAAAGGGcAGCUGAGCUdTsdT-3′ (SEQ ID NO: 577),

5′-UUCcAAAGGGcAGCUGAGCdTsdT-3′ (SEQ ID NO: 585),

5′-uAUUCcAAAGGGcAGCUGAdTsdT-3′ (SEQ ID NO: 593), and

5′-UuAUUCcAAAGGGcAGCUGdTsdT-3′ (SEQ ID NO: 601),

wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

16. The dsRNA of claim 1 , wherein the sense strand comprises the nucleotide sequence of any one of

5′-AGcucAGcuGcccuuuGGAdTsdT-3′ (SEQ ID NO: 576),

5′-GcucAGcuGcccuuuGGAAdTsdT-3′ (SEQ ID NO: 584),

5′-ucAGcuGcccuuuGGAAuAdTsdT-3′ (SEQ ID NO: 592), or

5′-cAGcuGcccuuuGGAAuAAdTsdT-3′ (SEQ ID NO: 600),

wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

17. The dsRNA of claim 1 ,

wherein the antisense strand comprises the nucleotide sequence of 5′ UCcAAAGGGcAGCUGAGCUdTsdT-3′ (SEQ ID NO: 577) and the sense strand comprises the nucleotide sequence of 5′-AGcucAGcuGcccuuuGGAdTsdT-3′ (SEQ ID NO: 576);

wherein the antisense strand comprises the nucleotide sequence of 5′-UUCcAAAGGGcAGCUGAGCdTsdT-3′ (SEQ ID NO: 585) and the sense strand comprises the nucleotide sequence of 5′-GcucAGcuGcccuuuGGAAdTsdT-3′ (SEQ ID NO: 584);

wherein the antisense strand comprises the nucleotide sequence of 5′-uAUUCcAAAGGGcAGCUGAdTsdT-3′ (SEQ ID NO: 593) and the sense strand comprises the nucleotide sequence of 5′-ucAGcuGcccuuuGGAAuAdTsdT-3′ (SEQ ID NO: 592); or

wherein the antisense strand comprises the nucleotide sequence of 5′-UuAUUCcAAAGGGcAGCUGdTsdT-3′ (SEQ ID NO: 601) and the sense strand comprises the nucleotide sequence of 5′-cAGcuGcccuuuGGAAuAAdTsdT-3′ (SEQ ID NO: 600), and

wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

18. The dsRNA agent of claim 1 , wherein at least one end of the dsRNA is blunt.

19. The dsRNA agent of claim 1 , wherein the dsRNA comprises a duplex region between 15-30 base pairs.

20. The dsRNA agent of claim 4 , wherein the ligand is a cell or tissue targeting group chosen from a lectin, a glycoprotein, a lipid, or an antibody that binds to a specified cell type.

21. The dsRNA agent of claim 4 , wherein the ligand is a multivalent galactose, N-acetyl-galactosamine, an N-acetyl-galacosamine multivalent mannose, or a cholesterol.

22. The dsRNA agent of claim 1 , wherein the sense strand consists of a nucleotide sequence selected from the group consisting of

5′-AGcucAGcuGcccuuuGGAdTsdT-3′ (SEQ ID NO: 576),

5′-GcucAGcuGcccuuuGGAAdTsdT-3′ (SEQ ID NO: 584),

5′-ucAGcuGcccuuuGGAAuAdTsdT-3′ (SEQ ID NO: 592), and

5′-cAGcuGcccuuuGGAAuAAdTsdT-3′ (SEQ ID NO: 600),

wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.

23. The dsRNA agent of claim 22 , wherein the antisense strand consists of a nucleotide sequence selected from the group consisting of

5′-UCcAAAGGGcAGCUGAGCUdTsdT-3′ (SEQ ID NO: 577),

5′-UUCcAAAGGGcAGCUGAGCdTsdT-3′ (SEQ ID NO: 585),

5′-uAUUCcAAAGGGcAGCUGAdTsdT-3′ (SEQ ID NO: 593), and

5′-UuAUUCcAAAGGGcAGCUGdTsdT-3′ (SEQ ID NO: 601); and

wherein the sense strand consists of a nucleotide sequence selected from the group consisting of

5′-AGcucAGcuGcccuuuGGAdTsdT-3′ (SEQ ID NO: 576),

5′-GcucAGcuGcccuuuGGAAdTsdT-3′ (SEQ ID NO: 584),

5′-ucAGcuGcccuuuGGAAuAdTsdT-3′ (SEQ ID NO: 592), and

5′-cAGcuGcccuuuGGAAuAAdTsdT-3′ (SEQ ID NO: 600),

wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.