Claims for Patent: 10,208,307

Summary for Patent: 10,208,307

Title:	Transthyretin (TTR) iRNA compositions and methods of use thereof for treating or preventing TTR-associated diseases
Abstract:	The present invention provides iRNA agents, e.g., double stranded iRNA agents, that target the transthyretin (TTR) gene and methods of using such iRNA agents for treating or preventing TTR-associated diseases.
Inventor(s):	Zimmermann; Tracy (Winchester, MA), Chan; Amy (Tewksbury, MA), Jadhav; Vasant (Sharon, MA), Maier; Martin (Belmont, MA), Rajeev; Kallanthottathil G. (Wayland, MA)
Assignee:	Alnylam Pharmaceuticals, Inc. (Cambridge, MA)
Application Number:	15/221,651
Patent Claims:	1. A double stranded ribonucleic acid (RNAi) agent that inhibits expression of transthyretin (TTR) in a cell, comprising a sense strand differing by no more than 4 modified nucleotides from the nucleotide sequence of 5'-usgsggauUfuCfAfUfguaaccaaga-3' (SEQ ID NO: 10) and an antisense strand differing by no more than 4 modified nucleotides from the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7), wherein a, c, g, and u are 2'-O-methyl (2'-OMe) A, C, G, and U; Af, Cf, Gf, and Uf are 2'-fluoro A, C, G, and U; and s is a phosphorothioate linkage. 2. The double stranded RNAi agent of claim 1, wherein the sense strand differs by no more than 3 modified nucleotides from the nucleotide sequence of 5'-usgsggauUfuCfAfUfguaaccaaga-3' (SEQ ID NO: 10) and the antisense strand differs by no more than 3 modified nucleotides from the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7). 3. The double stranded RNAi agent of claim 1, wherein the sense strand differs by no more than 2 modified nucleotides from the nucleotide sequence of 5'-usgsggauUfuCfAfUfguaaccaaga-3' (SEQ ID NO: 10) and the antisense strand differs by no more than 2 modified nucleotides from the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7). 4. The double stranded RNAi agent of claim 1, wherein the sense strand differs by no more than 1 modified nucleotide from the nucleotide sequence of 5'-usgsggauUfuCfAfUfguaaccaaga-3' (SEQ ID NO: 10) and the antisense strand differs by no more than 1 modified nucleotide from the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7). 5. The double stranded RNAi agent of claim 1, wherein the sense strand comprises the nucleotide sequence 5'-usgsggauUfuCfAfUfguaaccaaga-3' (SEQ ID NO: 10) and the antisense strand comprises the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7). 6. The double stranded RNAi agent of claim 1, wherein the sense strand consists of the nucleotide sequence 5'-usgsggauUfuCfAfUfguaaccaaga-3' (SEQ ID NO: 10) and the antisense strand consists of the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7). 7. The double stranded RNAi agent of claims 1-6, further comprising at least one ligand. 8. The double stranded RNAi agent of claims 1-6, wherein the sense strand is conjugated to at least one ligand. 9. The double stranded RNAi agent of claim 8, wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker. 10. The double stranded RNAi agent of claim 8, wherein the ligand is ##STR00017## 11. The double stranded RNAi agent of claim 8, wherein the ligand is attached to the 3' end of the sense strand. 12. The double stranded RNAi agent of claim 11, wherein the RNAi agent is conjugated to the ligand as shown in the following schematic ##STR00018## wherein X is O or S. 13. The double stranded RNAi agent of claim 12, wherein the X is O. 14. An isolated cell containing the double stranded RNAi agent of claim 1. 15. A vector containing the double stranded RNAi agent of claim 1. 16. An isolated cell comprising the vector of claim 15. 17. A pharmaceutical composition comprising the double stranded RNAi agent of claim 1. 18. The pharmaceutical composition of claim 17, comprising an unbuffered solution. 19. The pharmaceutical composition of claim 18, wherein the unbuffered solution is saline or water. 20. The pharmaceutical composition of claim 17, comprising a buffered solution. 21. The pharmaceutical composition of claim 20, wherein the buffered solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof. 22. A double stranded ribonucleic acid (RNAi) agent, comprising a sense strand and an antisense strand, wherein the sense strand comprises the nucleotide sequence 5'-usgsggauUfuCfAfUfguaaccaagaL96-3' (SEQ ID NO: 10) and the antisense strand comprises the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7), wherein a, c, g, and u are 2'-O-methyl (2'-OMe) A, C, G, and U; Af, Cf, Gf, and Uf are 2'-fluoro A, C, G, and U; s is a phosphorothioate linkage; and L96 is N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol. 23. A pharmaceutical composition comprising the double stranded RNAi agent of claim 22. 24. The pharmaceutical composition of claim 23, comprising an unbuffered solution. 25. The pharmaceutical composition of claim 24, wherein the unbuffered solution is saline or water. 26. The pharmaceutical composition of claim 23, comprising a buffered solution. 27. The pharmaceutical composition of claim 26, wherein the buffered solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof. 28. An isolated cell containing the double stranded RNAi agent of claim 22. 29. A vector containing the double stranded RNAi agent of claim 22. 30. A double stranded ribonucleic acid (RNAi) agent comprising a sense strand and an antisense strand, wherein the sense strand consists of the nucleotide sequence 5'-usgsggauUfuCfAfUfguaaccaagaL96-3' (SEQ ID NO: 10) and the antisense strand consists of the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7), wherein a, c, g, and u are 2'-O-methyl (2'-OMe) A, C, G, and U; Af, Cf, Gf, and Uf are 2'-fluoro A, C, G, and U; s is a phosphorothioate linkage; and L96 is N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol. 31. A pharmaceutical composition comprising the double stranded RNAi agent of claim 30. 32. The pharmaceutical composition of claim 31, comprising an unbuffered solution. 33. The pharmaceutical composition of claim 32, wherein the unbuffered solution is saline or water. 34. The pharmaceutical composition of claim 31, comprising a buffered solution. 35. The pharmaceutical composition of claim 34, wherein the buffered solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof. 36. An isolated cell containing the double stranded RNAi agent of claim 30. 37. A vector containing the double stranded RNAi agent of claim 30. 38. A method of inhibiting transthyretin (TTR) expression in a cell, the method comprising: contacting the cell with the double stranded RNAi agent of claim 1, the vector of claim 15 or the pharmaceutical composition of claim 17, thereby inhibiting expression of the TTR gene in the cell. 39. The method of claim 38, wherein the cell is within a subject. 40. The method of claim 39, wherein the subject is a human. 41. The method of claim 40, wherein the subject suffers from a TTR-associated disease. 42. A method of treating a subject suffering from a TTR-associated disease, comprising administering to the subject a therapeutically effective amount of a double stranded ribonucleic acid (RNAi), wherein the double stranded RNAi agent comprises a sense strand differing by no more than 4 modified nucleotides from the nucleotide sequence of 5'-usgsggauUfuCfAfUfguaaccaaga-3' (SEQ ID NO: 10) and an antisense strand differing by no more than 4 modified nucleotides from the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7), wherein a, c, g, and u are 2'-O-methyl (2'-OMe) A, C, G, and U; Af, Cf, Gf, and Uf are 2'-fluoro A, C, G, and U; and s is a phosphorothioate linkage, thereby treating the subject. 43. The method of claim 42, wherein a Neuropathy Impairment Score (NIS) or a modified NIS (mNIS+7) is reduced, slowed, or arrested in the subject suffering from a TTR-associated disease. 44. The method of claim 42, wherein a 6-minute walk test (6MWT) is increased in the subject suffering from a TTR-associated disease. 45. The method of claim 42, wherein the subject is a human. 46. The method of claim 42, wherein said subject carries a TTR gene mutation that is associated with the development of a TTR-associated disease. 47. The method of claim 42, wherein said TTR-associated disease is selected from the group consisting of senile systemic amyloidosis (SSA), systemic familial amyloidosis, familial amyloidotic polyneuropathy (FAP), familial amyloidotic cardiomyopathy (FAC), leptomeningeal/Central Nervous System (CNS) amyloidosis, and hyperthyroxinemia. 48. The method of claim 42, wherein said double stranded RNAi agent is administered to said subject by an administration means selected from the group consisting of subcutaneous, intravenous, intramuscular, intrabronchial, intrapleural, intraperitoneal, intraarterial, lymphatic, cerebrospinal, and any combinations thereof. 49. The method of claim 42, wherein said double stranded RNAi agent is administered to said subject via subcutaneous administration. 50. The method of claim 49, wherein the subcutaneous administration is self administration. 51. The method of claim 50, wherein the self-administration is via a pre-filled syringe or auto-injector syringe. 52. The method of claim 42, wherein the double stranded RNAi agent is chronically administered to the subject. 53. The method of claim 42, wherein the sense strand differs by no more than 3 modified nucleotides from the nucleotide sequence of 5'-usgsggauUfuCfAfUfguaaccaaga-3' (SEQ ID NO: 10) and the antisense strand differs by no more than 3 modified nucleotides from the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7). 54. The method of claim 42, wherein the sense strand differs by no more than 2 modified nucleotides from the nucleotide sequence of 5'-usgsggauUfuCfAfUfguaaccaaga-3' (SEQ ID NO: 10) and the antisense strand differs by no more than 2 modified nucleotides from the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7). 55. The method of claim 42, wherein the sense strand differs by no more than 1 modified nucleotide from the nucleotide sequence of 5'-usgsggauUfuCfAfUfguaaccaaga-3' (SEQ ID NO: 10) and the antisense strand differs by no more than 1 modified nucleotide from the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7). 56. The method of claim 42, wherein the sense strand comprises the nucleotide sequence 5'-usgsggauUfuCfAfUfguaaccaaga-3' (SEQ ID NO: 10) and the antisense strand comprises the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7). 57. The method of claim 42, wherein the sense strand consists of the nucleotide sequence 5'-usgsggauUfuCfAfUfguaaccaaga-3' (SEQ ID NO: 10) and the antisense strand consists of the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7). 58. The method of claims 42 and 53-57, wherein the double stranded RNAi agent further comprises at least one ligand. 59. The method of claims 42 and 53-57, wherein the sense strand of the double stranded RNAi agent is conjugated to at least one ligand. 60. The method of claim 59, wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker. 61. The method of claim 59, wherein the ligand is ##STR00019## 62. The method of claim 59, wherein the ligand is attached to the 3' end of the sense strand. 63. The method of claim 62, wherein the double stranded RNAi agent is conjugated to the ligand as shown in the following schematic ##STR00020## wherein X is O or S. 64. The method of claim 63, wherein the X is 0. 65. A method of treating a subject suffering from a TTR-associated disease, comprising administering to the subject a therapeutically effective amount of a double stranded ribonucleic acid (RNAi) agent, wherein the double stranded RNAi agent comprises a sense strand and an antisense strand, wherein the sense strand comprises the nucleotide sequence 5'-usgsggauUfuCfAfUfguaaccaagaL96-3' (SEQ ID NO: 10) and the antisense strand comprises the nucleotide sequence 5'-usCfsuugGfuuAfcaugAfaAfucccasusc-3' (SEQ ID NO: 7) wherein a, c, g, and u are 2'-O-methyl (2'-OMe) A, C, G, and U; Af, Cf, Gf, and Uf are 2'-fluoro A, C, G, and U; s is a phosphorothioate linkage; and L96 is N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol, thereby treating the subject. 66. The method of claim 65, wherein a Neuropathy Impairment Score (NIS) or a modified NIS (mNIS+7) is reduced, slowed, or arrested in the subject suffering from a TTR-associated disease. 67. The method of claim 65, wherein a 6-minute walk test (6MWT) is increased in the subject suffering from a TTR-associated disease. 68. The method of claim 65, wherein the subject is a human. 69. The method of claim 65, wherein the subject carries a TTR gene mutation that is associated with the development of a TTR-associated disease. 70. The method of claim 65, wherein the TTR-associated disease is selected from the group consisting of senile systemic amyloidosis (SSA), systemic familial amyloidosis, familial amyloidotic polyneuropathy (FAP), familial amyloidotic cardiomyopathy (FAC), leptomeningeal/Central Nervous System (CNS) amyloidosis, and hyperthyroxinemia. 71. The method of claim 65, wherein the double stranded RNAi agent is administered to said subject by an administration means selected from the group consisting of subcutaneous, intravenous, intramuscular, intrabronchial, intrapleural, intraperitoneal, intraarterial, lymphatic, cerebrospinal, and any combinations thereof. 72. The method of claim 65, wherein the double stranded RNAi agent is administered to the subject via subcutaneous administration. 73. The method of claim 72, wherein the subcutaneous administration is self administration. 74. The method of claim 73, wherein the self-administration is via a pre-filled syringe or auto-injector syringe. 75. The method of claim 65, wherein the double stranded RNAi agent is chronically administered to the subject.