Claims for Patent: 8,741,866

Summary for Patent: 8,741,866

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:	13/410,262
Patent Claims:	1. A method of inhibiting TTR expression in a cell, the method comprising: (a) contacting the cell with a double-stranded ribonucleic acid (dsRNA) for inhibiting expression of transthyretin (TTR), wherein the dsRNA comprises a sense strand and an antisense strand, the 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 of the dsRNA is 19, 20, 21, 22, 23, or 24 nucleotides in length; 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. 2. The method of claim 1, wherein the method is performed in vitro. 3. The method of claim 1, wherein the method is performed in vivo. 4. The method of claim 1, wherein the region of complementarity consists of SEQ ID NO:170. 5. The method of claim 1, wherein the sense strand consists of SEQ ID NO:449 and the antisense strand consists of SEQ ID NO:450. 6. The method of claim 1, wherein the sense strand consists of SEQ ID NO:729 and the antisense strand consists of SEQ ID NO:730. 7. The method of claim 1, wherein the sense strand consists of SEQ ID NO:1009 and the antisense strand consists of SEQ ID NO:1010 and the DsRnA is formulated in a lipid formulation. 8. The method of claim 1, wherein each strand is 21 nucleotides in length. 9. The method of claim 1, wherein the dsRNA comprises at least one modified nucleotide. 10. The method of claim 1, wherein the dsRNA comprises at least one modified nucleotide chosen from the group 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, 2'-amino-modified nucleotide, 2'-alkyl-modified nucleotide, morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide. 11. The method of claim 1, wherein the dsRNA comprises at least one 2'-O-methyl modified nucleotide. 12. The method of claim 1, wherein the dsRNA is conjugated to a ligand. 13. The method of claim 1, wherein the dsRNA is formulated in a lipid formulation. 14. The method of claim 1, wherein the dsRNA is formulated in a lipid formulation comprising MC3. 15. The method of claim 1, wherein contacting the cell with the dsRNA results in at least one of the following: a) 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 the dsRNA concentration is 10 nM; b) about 74% inhibition of TTR mRNA expression as measured by a branched DNA assay, wherein the cell is a HepG2 cell or a Hep3B cell and the dsRNA concentration is 10 nM; c) the dsRNA has an IC50 of less than 10 pM in a HepG2 cell and the dsRNA concentration is 10 nM; d) the dsRNA has an ED50 of about 1 mg/kg; e) the dsRNA reduces TTR expression by 98.9% as measured by real-time PCR in a Hep3B cell and the dsRNA concentration is 0.1 nM; f) the dsRNA reduces TTR expression by 99.4% as measured by real-time PCR in a Hep3B cell and the dsRNA concentration is 10 nM. 16. 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 a double-stranded ribonucleic acid (dsRNA) for inhibiting expression of transthyretin (TTR), wherein the dsRNA comprises a sense strand and an antisense strand, the 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 of the dsRNA is 19, 20, 21, 22, 23, or 24 nucleotides in length, wherein the disorder is a transthyretin amyloidosis selected from the group consisting of familial amyloidotic polyneuropathy (FAP), familial amyloidotic cardiomyopathy (FAC), leptomeningeal/CNS amyloidosis, senile systemic amyloidosis (SSA) and senile cardiac amyloidosis (SCA). 17. The method of claim 16, wherein the dsRNA is administered to the human at about 0.01, 0.1, 0.3, 0.5, 1.0, 2.5, or 5.0 mg/kg. 18. The method of claim 16, wherein the dsRNA is administered to the human at about 1.0 mg/kg. 19. The method of claim 16, wherein the dsRNA is formulated in a lipid formulation. 20. The method of claim 16, wherein the dsRNA is formulated in a lipid formulation comprising MC3. 21. The method of claim 16, wherein the human has a liver disorder. 22. The method of claim 16, wherein the human is further provided a liver transplant. 23. The method of claim 16, wherein administration of the dsRNA results in at least one of the following: a) reduction of TTR mRNA by about 80% in human liver, wherein the therapeutic amount is 3 mg/kg; b) no immunostimulatory activity in the human as measured by IFN-alpha and TNF-alpha ELISA assays; c) reduction of liver TTR mRNA levels by about 97% or serum TTR protein levels by about 90%, wherein the therapeutic amound is 6 mg/kg; d) reduction of liver TTR mRNA levels and/or serum TTR protein levels up to 22 days, wherein therapeutic amount is 6 mg/kg or 3 mg/kg or e) suppression of serum TTR protein levels up to day 14 post-treatment wherein therapeutic amount is 1 mg/kg or 3 mg/kg. 24. The method of claim 16, wherein the dsRNA is formulated in a lipid formulation comprising MC3 and administration of the dsRNA results in at least one of the following: a) reduction of TTR mRNA levels by about 85 to 90% at a therapeutic amount of 0.3 mg/kg, relative to a PBC control group or b) reduction TTR mRNA levels by about 50% at a therapeutic amount 0.1 mg/kg, relative to a PBC control group or c) reduction TTR protein levels in a dose-dependent manner relative to a PBC control group as measured by a western blot or d) suppression of serum TTR protein levels up to day 14 post-treatment when the therapeutic amount is 1 mg/kg or 3 mg/kg. 25. The method of claim 1, wherein the sense strand consists of SEQ ID NO:1009 and the antisense strand consists of SEQ ID NO:1010 and the dsRNA is lipid formulated in a formulation comprising MC3. 26. The method of claim 16, wherein the sense strand consists of SEQ ID NO:1009 and the antisense strand consists of SEQ ID NO:1010 and the dsRNA is lipid formulated in a formulation comprising MC3. 27. A method of treating familial amyloidotic polyneuropathy (FAP) comprising administering to a human in need of such treatment a double-stranded ribonucleic acid (dsRNA) comprising a sense strand and an antisense strand, the sense strand consisting of SEQ ID NO:1009 and the antisense strand consisting of SEQ ID NO:1010, wherein the dsRNA is in a formulation comprising the cationic lipid MC3 and the dsRNA is administered at a dose of 0.3 mg/kg. 28. The method of claim 16, wherein the dsRNA is administered intraveneously. 29. The method of claim 6, wherein the dsRNA is formulated in a lipid formulation. 30. The method of claim 6, wherein the dsRNA is formulated in a lipid formulation comprising MC3.