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 comprising any one of the modified nucleotide sequences selected from the group consisting of SEQ ID NOS: 62-86, 112-127, and 144-147, and an antisense strand comprising any one of the modified nucleotide sequences selected from the group consisting of SEQ ID NOS: 87-111, 128-143, and 148-151, and wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.
2. The double stranded RNAi agent of claim 1 , wherein the sense strand and the antisense strand comprise the sense and antisense strand nucleotide sequences of a duplex selected from the group consisting of AD-63202, AD-64372, AD-64373, AD-64374, AD-64375, AD-64376, AD-64377, AD-64378, AD-64380, AD-64381, AD-64382, AD-64384, AD-64385, AD-64386, AD-64387, AD-64389, AD-64601, AD-64569, AD-64604, AD-64567, AD-60940, AD-64601, AD-65105, AD-65106, AD-65107, AD-65108, AD-65109, AD-65110, AD-65111, AD-65112, AD-61002, AD-66014, AD-66015, and AD-65189.
3. The double stranded RNAi agent of claim 1 , further comprising a ligand.
4. The double stranded RNAi agent of claim 3 , wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.
5. The double stranded RNAi agent of claim 3 , wherein the ligand is
6. The double stranded RNAi agent of claim 3 , wherein the ligand is attached to the 3′ end of the sense strand.
7. The double stranded RNAi agent of claim 6 , wherein the RNAi agent is conjugated to the ligand as shown in the following schematic
wherein X is O or S.
8. A pharmaceutical composition comprising the double stranded RNAi agent of claim 1 .
9. 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 8 ; 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 said cell is within a subject.
11. The method of claim 10 , wherein the subject is a human.
12. The method of claim 9 , 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%.
13. The method of claim 10 , 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%.
14. 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 8 , thereby treating the subject.
15. The method of claim 14 , wherein the subject is a human.
16. The method of claim 15 , wherein the human has hereditary hemochromatosis, β-thalassemia, or erythropoietic porphyria.
17. The method of claim 15 , wherein the human has a disorder associated with iron overload.
18. The method of claim 14 , wherein the double stranded RNAi agent is administered to the subject subcutaneously; or intravenously.
19. The method of claim 14 , further comprising administering an iron chelator to the subject.
20. The double stranded RNAi agent of claim 1 ,
wherein the sense strand comprises the nucleotide sequence 5′-csusgguaUfuUfCfCfuaggGfdTacaa-3′ (SEQ ID NO: 72) and the antisense strand comprises the nucleotide sequence 5′- usUfsguaCfccuaggaAfaUfaccagsasg-3′ (SEQ ID NO: 97),
wherein a, c, g, and u are 2′-O-methyladenosine-3′-phosphate, 2′-O-methylcytidine-3′-phosphate, 2′-O-methylguanosine-3′-phosphate, and 2′-O-methyluridine-3′-phosphate, respectively; Af, Cf, Gf, and Uf are 2′-fluoroadenosine-3′-phosphate, 2′-fluorocytidine-3′-phosphate, 2′-fluoroguanosine-3′-phosphate, and 2′-fluorouridine-3′-phosphate, respectively; s is a phosphorothioate linkage; and dT is 2′-deoxythymidine.