Compositions and methods for inhibiting expression of transthyretin

1. A composition comprising a double-stranded ribonucleic acid (dsRNA) for inhibiting expression of transthyretin (TTR) and a lipid formulation, wherein the dsRNA comprises a sense strand and an antisense strand comprising a region complementary to an mRNA encoding transthyretin (TTR), wherein the region of complementarity comprises SEQ ID NO:170 and each strand is 19, 20, 21, 22, 23, or 24 nucleotides in length, the lipid formulation comprising the lipid (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate (MC3).

2. The composition of claim 1 , wherein the sense strand comprises the nucleotide sequence of SEQ ID NO:169.

3. The composition of claim 1 , wherein the sense strand consists of SEQ ID NO:449 and the antisense strand consists of the nucleotide sequence of SEQ ID NO: 450.

4. The composition of claim 1 , wherein the sense strand consists of SEQ ID NO:729 and the antisense strand consists of SEQ ID NO:730.

5. The composition of claim 1 , wherein the sense strand consists of SEQ ID NO: 1009 and the antisense strand consists of SEQ ID NO:1010.

6. The composition of claim 1 , wherein each strand is 21 nucleotides in length.

7. The composition of claim 1 , wherein the dsRNA does not cleave a TTR mRNA between the adenine nucleotide at position 637 of SEQ ID NO:1331 and the guanine nucleotide at position 638 of SEQ ID NO:1331.

8. The composition of claim 1 , wherein the dsRNA cleaves a TTR mRNA between the guanine nucleotide at position 636 of SEQ ID NO:1331 and the adenine nucleotide at position 637 of SEQ ID NO:1331.

9. The composition of claim 1 , wherein the dsRNA anneals to a TTR mRNA between the guanine nucleotide at position 628 of SEQ ID NO:1331 and the uracil nucleotide at position 646 of SEQ ID NO: 1331.

10. The composition of claim 1 , wherein the antisense strand base pairs with the guanine at position 628 of SEQ ID NO:1331.

11. The composition of claim 1 , wherein the dsRNA comprises at least one modified nucleotide.

12. The composition of claim 1 , wherein the dsRNA comprises at least one modified nucleotide selected from the group consisting of: a 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group, a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide.

13. The composition of claim 1 , wherein the dsRNA comprises at least one 2′-O-methyl modified nucleotide.

14. The composition of claim 1 , wherein the lipid formulation further comprises distearoylphosphatidylcholine (DSPC).

15. The composition of claim 1 , wherein the lipid formulation further comprises cholesterol.

16. The composition of claim 1 , wherein the lipid formulation further comprises polyethyleneglycol (PEG) or PEG-DMG.

17. The composition of claim 1 , wherein the lipid formulation further comprises DSPC, cholesterol, and PEG or PEG-DMG.

18. The composition of claim 1 , wherein the lipid formulation comprises a MC3/DSPC/cholesterol/PEG-DMG ratio of 50/10/38.5/1.5 mol %.

19. The composition of claim 1 , wherein the dsRNA is conjugated to a ligand.

20. The composition of claim 1 , wherein administration of the composition to a cell results in about 95% inhibition of TTR mRNA expression as measured by a real time PCR assay, wherein the cell is a HepG2 cell or a Hep3B cell, and wherein the concentration of the dsRNA is 10 nM.

21. A cell containing the composition of claim 1 .

22. A method of inhibiting TTR expression in a cell, the method comprising:

(a) contacting the cell with the composition 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 TTR gene, thereby inhibiting expression of the TTR gene in the cell.

23. A method of treating a disorder mediated by TTR expression comprising administering to a human in need of such treatment a therapeutically effective amount of the composition of claim 1 .

24. The method of claim 23 , wherein the human has transthyretin amyloidosis, familial amyloidotic polyneuropathy (FAP), familial amyloidotic cardiomyopathy (FAC), leptomeningeal/CNS amyloidosis, senile systemic amyloidosis (SSA), senile cardiac amyloidosis (SCA), a liver disorder, or is further provided a liver transplant.

25. The composition of claim 5 , wherein the lipid formulation comprises a MC3/DSPC/cholesterol/PEG-DMG ratio of 50/10/38.5/1.5 mol %.

26. A method of inhibiting TTR expression in a cell, the method comprising:

(a) contacting the cell with the composition of claim 5 ; and

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

27. A method of treating a disorder mediated by TTR expression comprising administering to a human in need of such treatment a therapeutically effective amount of the composition of claim 5 .

28. The method of claim 27 , wherein the human has transthyretin amyloidosis, familial amyloidotic polyneuropathy (FAP), familial amyloidotic cardiomyopathy (FAC), leptomeningeal/CNS amyloidosis, senile systemic amyloidosis (SSA), senile cardiac amyloidosis (SCA), a liver disorder, or is further provided a liver transplant.

29. The method of claim 27 , wherein the human has familial amyloidotic polyneuropathy (FAP).

30. The method of claim 26 , wherein the method is performed in vitro.

31. The method of claim 26 , wherein the method is performed in vivo.

32. A method of inhibiting TTR expression in a cell, the method comprising:

(a) contacting the cell with the composition of claim 25 ; and

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

33. The method of claim 32 , wherein the method is performed in vitro.

34. The method of claim 32 , wherein the method is performed in vivo.

35. A method of treating a disorder mediated by TTR expression comprising administering to a human in need of such treatment a therapeutically effective amount of the composition of claim 25 .

36. The method of claim 35 , wherein the human has transthyretin amyloidosis, familial amyloidotic polyneuropathy (FAP), familial amyloidotic cardiomyopathy (FAC), leptomeningeal/CNS amyloidosis, senile systemic amyloidosis (SSA), senile cardiac amyloidosis (SCA), a liver disorder, or is further provided a liver transplant.

37. The method of claim 35 , wherein the human has familial amyloidotic polyneuropathy (FAP).