Claims for Patent: 9,234,196

Summary for Patent: 9,234,196

Title:	Compositions and methods for inhibiting expression of transthyretin
Abstract:	The invention relates to a double-stranded ribonucleic acid (dsRNA) targeting a transthyretin (TTR) gene, and methods of using the dsRNA to inhibit expression of TTR.
Inventor(s):	Sah; Dinah Wen-Yee (Boston, MA), Hinkle; Gregory (Plymouth, MA), Alvarez; Rene (Boxborough, MA), Milstein; Stuart (Cambridge, MA), Chen; Qingmin (Lincoln, MA)
Assignee:	Alnylam Pharmaceuticals, Inc. (Cambridge, MA)
Application Number:	14/220,829
Patent Claims:	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).