TMPRSS6 iRNA compositions and methods of use thereof
The invention relates to RNAi agents, e.g., double-stranded RNAi agents, targeting the TMPRSS6 gene, and methods of using such RNAi agents to inhibit expression of TMPRSS6 and methods of treating subjects having a TMPRSS6 associated disorder, e.g., an iron overload associated disorder, such as β-thalassemia or hemochromatosis.
1. A double stranded RNAi agent for inhibiting expression of TMPRSS6 in a cell, wherein said double stranded RNAi agent comprises a sense strand and an antisense strand forming a duplex,
wherein the sense strand comprises the nucleotide sequence 5′-csusggudAsuuucdCuaggg(Tgn)acaa-3′ (SEQ ID NO: 126) and the antisense strand comprises the nucleotide sequence 5′-usUsguacccudAggadAauaccagsasg-3′ (SEQ ID NO: 142),
wherein a, c, g, and u are 2′-O-methyl (2′-OMe) A, C, G, and U; U is uridine-3′-phosphate; s is a phosphorothioate linkage; dA is 2′-deoxyadenosine; dC is 2′-deoxycytosine; and (Tgn) is thymidine-glycol nucleic acid (GNA)S-Isomer;
wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length; and
wherein the double stranded RNAi agent is conjugated to a ligand.
2. The double stranded RNAi agent of claim 1 , wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.
3. The double stranded RNAi agent of claim 1 , wherein the ligand is
4. The double stranded RNAi agent of claim 1 , wherein the ligand is attached to the 3′ end of the sense strand.
5. The double stranded RNAi agent of claim 4 , wherein the RNAi agent is conjugated to the ligand as shown in the following schematic
wherein X is O or S.
6. A pharmaceutical composition comprising the double stranded RNAi agent of claim 1 .
7. A method of inhibiting TMPRSS6 expression in a cell, the method comprising:
(a) contacting the cell with the double stranded RNAi agent of claim 1 , or the pharmaceutical composition of claim 6 ; 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.
8. The method of claim 7 , wherein said cell is within a subject.
9. The method of claim 8 , wherein the subject is a human.
10. The method of claim 7 , wherein TMPRSS6 expression is inhibited by at least about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 98% or about 100%.
11. The method of claim 8 , wherein serum hepcidin concentration in said subject is increased by at least about 10%; and/or wherein serum iron concentration in said subject is decreased by at least about 20%; and/or wherein a percent transferrin saturation in said subject is decreased by at least about 20%.
12. A method of treating a subject having a TMPRSS6 associated disorder, comprising administering to the subject a therapeutically effective amount of the double stranded RNAi agent of claim 1 , or the pharmaceutical composition of claim 6 , thereby treating the subject.
13. The method of claim 12 , wherein the subject is a human.
14. The method of claim 13 , wherein the human has hereditary hemochromatosis, β-thalassemia, or erythropoietic porphyria.
15. The method of claim 13 , wherein the human has a disorder associated with iron overload.
16. The method of claim 12 , wherein the double stranded RNAi agent is administered to the subject subcutaneously; or intravenously.
17. The method of claim 12 , further comprising administering an iron chelator to the subject.