Transmembrane protease, serine 6 (TMPRSS6) iRNA compositions and methods of use thereof
The present invention relates to RNAi agents, e.g., double stranded RNA (dsRNA) agents, targeting the Transmembrane protease, serine 6 (TMPRSS6) gene. The invention also relates to methods of using such RNAi agents to inhibit expression of a TMPRSS6 gene and to methods of preventing and treating a TMPRSS6-associated disorder, e.g., a disorder associated with iron overload and/or a disorder of ineffective erythropoiesis, e.g., hereditary hemochromatosis, β-thalassemia (e.g., β-thalassemia major and β-thalassemia intermedia), polycythemia vera, myelodysplastic syndrome, congenital dyserythropoietic anemias, pyruvate kinase deficiency, erythropoietic porphyria, Parkinson's Disease, Alzheimer's Disease or Friedreich's Ataxia.
1. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of Transmembrane protease, serine 6 (TMPRSS6) in a cell, or a pharmaceutically acceptable salt thereof, comprising a sense strand and an antisense strand forming a double stranded region,
wherein the nucleotide sequence of the sense strand differs by no more than 4 bases from the nucleotide sequence 5′-asgscugcccUfUfUfggaauaaagu-3′ (SEQ ID NO:395) and the nucleotide sequence of the antisense strand differs by no more than 4 bases from the nucleotide sequence 5′-asdCsuudTadTuccadAaGfggcagcusgsa-3′ (SEQ ID NO:521),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Gf and Uf are 2′-deoxy-2′-fluoro (2′-F) G and U, respectively; dC, dA, and dT are 2′-deoxy C, A, and T, respectively; and s is a phosphorothioate linkage, and
wherein the dsRNA agent is conjugated to a ligand.
2. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1 ,
wherein the nucleotide sequence of the sense strand differs by no more than 3 bases from the nucleotide sequence 5′-asgscugcccUfUfUfggaauaaagu-3′ (SEQ ID NO:395) and the nucleotide sequence of the antisense strand differs by no more than 3 bases from the nucleotide sequence 5′-asdCsuudTadTuccadAaGfggcagcusgsa-3′ (SEQ ID NO:521).
3. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1 ,
wherein the nucleotide sequence of the sense strand differs by no more than 2 bases from the nucleotide sequence 5′-asgscugcccUfUfUfggaauaaagu-3′ (SEQ ID NO:395) and the nucleotide sequence of the antisense strand differs by no more than 2 bases from the nucleotide sequence 5′-asdCsuudTadTuccadAaGfggcagcusgsa-3′(SEQ ID NO:521).
4. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1 ,
wherein the nucleotide sequence of the sense strand differs by no more than 1 base from the nucleotide sequence 5′-asgscugcccUfUfUfggaauaaagu-3′ (SEQ ID NO:395) and the nucleotide sequence of the antisense strand differs by no more than 1 base from the nucleotide sequence 5′-asdCsuudTadTuccadAaGfggcagcusgsa-3′ (SEQ ID NO:521).
5. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1 ,
wherein the nucleotide sequence of the sense strand comprises the nucleotide sequence 5′-asgscugcccUfUfUfggaauaaagu-3′ (SEQ ID NO:395) and the nucleotide sequence of the antisense strand comprises the nucleotide sequence 5′-asdCsuudTadTuccadAaGfggcagcusgsa-3′ (SEQ ID NO:521).
6. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1 ,
wherein the nucleotide sequence of the sense strand consists of the nucleotide sequence 5′-asgscugcccUfUfUfggaauaaagu-3′ (SEQ ID NO:395) and the nucleotide sequence of the antisense strand consists of the nucleotide sequence 5′-asdCsuudTadTuccadAaGfggcagcusgsa-3′ (SEQ ID NO:521).
7. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1 , wherein the ligand is conjugated to the 3′ end of the sense strand of the dsRNA agent.
8. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
9. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 8 , wherein the ligand is one or more GalNAc derivatives attached through a monovalent, bivalent, or trivalent linker.
10. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 9 , wherein the ligand is
11. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 10 , wherein the dsRNA agent is conjugated to the ligand as shown in the following schematic
and, wherein X is O or S.
12. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 11 , wherein X is O.
13. An isolated cell containing the dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1 .
14. A pharmaceutical composition for inhibiting expression of a gene encoding Transmembrane protease, serine 6 (TMPRSS6) comprising the dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1 .
15. The pharmaceutical composition of claim 14 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is in an unbuffered solution.
16. The pharmaceutical composition of claim 15 , wherein the unbuffered solution is saline or water.
17. The pharmaceutical composition of claim 14 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is in a buffer solution.
18. The pharmaceutical composition of claim 17 , wherein the buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof.
19. The pharmaceutical composition of claim 18 , wherein the buffer solution is phosphate buffered saline (PBS).
20. A composition, or a pharmaceutically acceptable salt thereof, comprising a sense strand and an antisense strand,
wherein the sense strand comprises the nucleotide sequence 5′-asgscugcccUfUfUfggaauaaaguL96-3′ (SEQ ID NO:395) and the antisense strand comprises the nucleotide sequence 5′-asdCsuudTadTuccadAaGfggcagcusgsa-3′ (SEQ ID NO:521),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Gf and Uf are 2′-deoxy-2′-fluoro (2′-F) G and U, respectively; dC, dA, and dT are 2′-deoxy C, A, and T, respectively; and s is a phosphorothioate linkage, and
wherein L96 is a ligand conjugated to the 3′-end of the sense strand as shown in the following schematic
wherein X is O.
21. The composition, or a pharmaceutically acceptable salt thereof, of claim 20 , which is in a sodium salt form.
22. An isolated cell containing the composition, or a pharmaceutically acceptable salt thereof, of claim 20 .
23. A pharmaceutical composition comprising the composition, or a pharmaceutically acceptable salt thereof, of claim 20 .
24. A composition, or a pharmaceutically acceptable salt thereof, comprising a sense strand and an antisense strand,
wherein the sense strand consists of the nucleotide sequence 5′-asgscugcccUfUfUfggaauaaaguL96-3′ (SEQ ID N0:395) and the antisense strand consists of the nucleotide sequence 5′-asdCsuudTadTuccadAaGfggcagcusgsa-3′ (SEQ ID NO:521),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Gf and Uf are 2′-deoxy-2′-fluoro (2′-F) G and U, respectively; dC, dA, and dT are 2′-deoxy C, A, and T, respectively; and s is a phosphorothioate linkage, and
wherein L96 is a ligand conjugated to the 3′-end of the sense strand as shown in the following schematic
and wherein X is O.
25. The composition, or a pharmaceutically acceptable salt thereof, of claim 24 , which is in a sodium salt form.
26. An isolated cell containing the composition, or a pharmaceutically acceptable salt thereof, of claim 24 .
27. A pharmaceutical composition comprising the composition, or a pharmaceutically acceptable salt thereof, of claim 24 .
28. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of Transmembrane protease, serine 6 (TMPRSS6) in a cell, or a pharmaceutically acceptable salt thereof, comprising a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand comprises the nucleotide sequence 5′-asgscugcccUfUfUfggaauaaagu-3′ (SEQ ID N0:395) and the antisense strand comprises the nucleotide sequence 5′-asdCsuudTadTuccadAaGfggcagcusgsa-3′ (SEQ ID NO:521),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Gf and Uf are 2′-deoxy-2′-fluoro (2′-F) G and U, respectively; dC, dA, and dT are 2′-deoxy C, A, and T, respectively; and s is a phosphorothioate linkage, and
wherein the dsRNA agent is conjugated to a ligand.
29. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 28 , which is in a sodium salt form.
30. A pharmaceutical composition comprising the dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 28 .
31. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1 , which is in a sodium salt form.
32. The pharmaceutical composition of claim 23 , wherein the composition, or a pharmaceutically acceptable salt thereof, is in an unbuffered solution.
33. The pharmaceutical composition of claim 32 , wherein the unbuffered solution is saline or water.
34. The pharmaceutical composition of claim 23 , wherein the composition, or a pharmaceutically acceptable salt thereof, is in a buffer solution.
35. The pharmaceutical composition of claim 34 , wherein the buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof.
36. The pharmaceutical composition of claim 35 , wherein the buffer solution is phosphate buffered saline (PBS).
37. The pharmaceutical composition of claim 27 , wherein the composition, or a pharmaceutically acceptable salt thereof, is in an unbuffered solution.
38. The pharmaceutical composition of claim 37 , wherein the unbuffered solution is saline or water.
39. The pharmaceutical composition of claim 27 , wherein the composition, or a pharmaceutically acceptable salt thereof, is in a buffer solution.
40. The pharmaceutical composition of claim 39 , wherein the buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof.
41. The pharmaceutical composition of claim 40 , wherein the buffer solution is phosphate buffered saline (PBS).
42. The pharmaceutical composition of claim 30 , wherein the composition, or a pharmaceutically acceptable salt thereof, is in an unbuffered solution.
43. The pharmaceutical composition of claim 42 , wherein the unbuffered solution is saline or water.
44. The pharmaceutical composition of claim 30 , wherein the composition, or a pharmaceutically acceptable salt thereof, is in a buffer solution.
45. The pharmaceutical composition of claim 44 , wherein the buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof.
46. The pharmaceutical composition of claim 45 , wherein the buffer solution is phosphate buffered saline (PBS).