Claims for Patent: 10,125,369

Summary for Patent: 10,125,369

Title:	PCSK9 iRNA compositions and methods of use thereof
Abstract:	The invention relates to RNAi agents, e.g., double-stranded RNAi agents, targeting the PCSK9 gene, and methods of using such RNAi agents to inhibit expression of PCSK9 and methods of treating subjects having a lipid disorder, such as a hyperlipidemia.
Inventor(s):	Borodovsky; Anna (Melrose, MA), Rajeev; Kallanthottathil G. (Wayland, MA), Fitzgerald; Kevin (Brookline, MA), Frank-Kamenetsky; Maria (Brookline, MA), Querbes; William (Boston, MA), Maier; Martin (Belmont, MA), Charisse; Klaus (Acton, MA), Kuchimanchi; Satyanarayana (Acton, MA), Manoharan; Muthiah (Weston, MA), Milstein; Stuart (Arlington, MA)
Assignee:	Alnylam Pharmaceuticals, Inc. (Cambridge, MA)
Application Number:	14/650,128
Patent Claims:	1. A double stranded RNAi agent that inhibits the expression of Proprotein convertase subtilisin kexin 9 (PCSK9) in a cell, wherein said double stranded RNAi agent comprises a sense strand complementary to an antisense strand forming a double stranded region, wherein said antisense strand comprises a region complementary to part of an mRNA encoding PCSK9, wherein each strand is independently about 14 to about 30 nucleotides in length, wherein said antisense strand comprises at least 19 contiguous nucleotides of the nucleotide sequence 5'-ACAAAAGCAAAACAGGUCUAG-3' (SEQ ID NO: 412) and said double stranded RNAi agent is represented by formula (III): TABLE-US-00037 sense: 5' n.sub.p-N.sub.a-(X X X).sub.i-N.sub.b-Y Y Y-N.sub.b-(Z Z Z).sub.j-N.sub.a-n.sub.q 3' antisense: 3' n.sub.p'-N.sub.a'-(X'X'X').sub.k-N.sub.b'-Y'Y'Y'-N.sub.b'-(Z'Z'Z').sub- .l- N.sub.a'-n.sub.q' 5' (III) wherein: j, k, and l are each independently 0 or 1; p, p', q, and q' are each independently 0-6; each N.sub.a and each N.sub.a' independently represents an oligonucleotide sequence comprising 0-25 nucleotides, 2-20 of which are modified nucleotides, each sequence comprising at least two differently modified nucleotides, wherein the modified nucleotides are each independently selected from the group consisting of 2'-O-methyl, 2'-fluoro, and 2'-deoxythymidine (dT); each N.sub.b and each N.sub.b' independently represents an oligonucleotide sequence comprising 0-10 nucleotides, 1-10 of which are modified nucleotides, wherein the modified nucleotides each independently comprise a nucleotide modification selected from the group consisting of 2'-O-methyl, 2'-fluoro, and 2'-deoxythymidine (dT); wherein the double stranded RNAi agent comprises at least one phosphorothioate or methylphosphonate internucleotide linkage; n.sub.p, n.sub.p', n.sub.q, and n.sub.q', each of which may or may not be present, each independently represents an overhang nucleotide; XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent one motif of three identical modifications on three consecutive nucleotides, wherein XXX is complementary to X'X'X', YYY is complementary to Y'Y'Y', and ZZZ is complementary to Z'Z'Z'; and wherein the sense strand is conjugated to at least one ligand which is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker. 2. The double stranded RNAi agent of claim 1, wherein the YYY motif occurs at or near the cleavage site of the sense strand. 3. The double stranded RNAi agent of claim 1, wherein the Y'Y'Y' motif occurs at the 11, 12 and 13 positions of the antisense strand from the 5'-end. 4. The double stranded RNAi agent of claim 1, wherein the modifications on the XXX nucleotides are different than the modifications on the X'X'X' nucleotides, the modifications on the YYY nucleotides are different than the modifications on the Y'Y'Y' nucleotides, and the modifications on the ZZZ nucleotides are different than the modifications on the Z'Z'Z' nucleotides. 5. The double stranded RNAi agent of claim 1, wherein the modifications on the YYY nucleotides are different than the modifications on the Y'Y'Y' nucleotides. 6. The double stranded RNAi agent of claim 1, wherein p'>0 and at least one np' is linked to a neighboring nucleotide via a phosphorothioate linkage. 7. The double stranded RNAi agent of claim 1, wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 5'-terminus of the sense strand; the 5'-terminus of the antisense strand; or at both the 5'-terminus of the sense strand and 5'-terminus of the antisense one strand. 8. The double stranded RNAi agent of claim 7, wherein the antisense strand comprises two phosphorothioate internucleotide linkages between the 3'-end three terminal nucleotides and two phosphorothioate internucleotide linkages between the 5' end three terminal nucleotides; and wherein the sense strand comprises two phosphorothioate internucleotide linkages between the 5' end three terminal nucleotides. 9. The double stranded RNAi agent of claim 1, wherein the double-stranded region is 15-30 nucleotide pairs in length. 10. The double stranded RNAi agent of claim 1, wherein the double-stranded region is 17-23 nucleotide pairs in length. 11. The double stranded RNAi agent of claim 1, wherein the double-stranded region is 17-25 nucleotide pairs in length. 12. The double stranded RNAi agent of claim 1, wherein the double-stranded region is 23-27 nucleotide pairs in length. 13. The double stranded RNAi agent of claim 1, wherein the double-stranded region is 19-21 nucleotide pairs in length. 14. The double stranded RNAi agent of claim 1, wherein the double-stranded region is or 21-23 nucleotide pairs in length. 15. The double stranded RNAi agent of claim 1, wherein each strand has 15-30 nucleotides. 16. The double stranded RNAi agent of claim 1, wherein the sense strand has a total of 21 nucleotides and the antisense strand has a total of 23 nucleotides. 17. The double stranded RNAi agent of claim 1, wherein the region complementary to part of an mRNA encoding PCSK9 comprises the nucleotide sequence of 5'-ACAAAAGCAAAACAGGUCUAGAA-3'(SEQ ID NO:1666). 18. The double stranded RNAi agent of claim 1, wherein the sense strand comprises the nucleotide sequence of 5'-CUAGACCUGUTUUGCUUUUGU-3' (SEQ ID NO:1665) and the antisense strand comprises the nucleotide sequence of 5'-ACAAAAGCAAAACAGGUCUAGAA-3'(SEQ ID NO:1666). 19. The double stranded RNAi agent of claim 1, wherein the antisense strand comprises the nucleotide sequence 5'-ACAAAAGCAAAACAGGUCUAG-3' (SEQ ID NO: 412) and the sense strand comprises the nucleotide sequence 5'-AGACCUGUUUUGCUUUUGU-3' (SEQ ID NO: 191). 20. The double stranded RNAi agent of claim 1, wherein the antisense strand consists of the nucleotide sequence 5'-ACAAAAGCAAAACAGGUCUAGAA-3'(SEQ ID NO:1666) and the sense strand consists of the nucleotide sequence 5'-CUAGACCUGUTUUGCUUUUGU-3' (SEQ ID NO: 1665). 21. The double stranded RNAi agent of claim 1, wherein the sense strand comprises the nucleotide sequence of 5'-csusagacCfuGfudTuugcuuuugu-3' (SEQ ID NO:1657) and the antisense strand comprises the nucleotide sequence of 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3' (SEQ ID NO:1663), 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; dT is 2'-deoxythymidine; and s is a phosphorothioate linkage. 22. The double stranded RNAi agent of claim 1, wherein the double stranded RNAi agent comprises: (a) an antisense strand consisting of the nucleotide sequence 5'-aCfaAfaAfgCfaAfaacAfgGfuCfuAfgsAfsa-3' (SEQ ID NO: 1151) and a sense strand consisting of the nucleotide sequence 5'-CfuAfgAfcCfuGfUfUfuUfgCfuUfuUfgUf-3' (SEQ ID NO: 600); (b) an antisense strand consisting of the nucleotide sequence 5'-aCfaAfAfAfgCfaAfaacAfgGfuCfuAfgsAfsa-3' (SEQ ID NO: 1246) and a sense strand consisting of the nucleotide sequence 5'-CfuAfgAfcCfuGfUfUfuUfgCfuuuUfgUf-3' (SEQ ID NO: 695); (c) an antisense strand consisting of the nucleotide sequence 5'-aCfaaaAfgCfaAfaacAfgGfuCfuAfgsAfsa-3' (SEQ ID NO: 1253) and a sense strand consisting of the nucleotide sequence 5'-CfuAfgAfcCfuGfUfUfuUfgCfuUfUfUfgUf-3' (SEQ ID NO: 702); (d) an antisense strand consisting of the nucleotide sequence 5'-aCfaAfAfAfgCfaAfaacAfgGfuCfusAfsg-3' (SEQ ID NO: 1263) and a sense strand consisting of the nucleotide sequence 5'-AfgAfcCfuGfUfUfuUfgCfuuuUfgUf-3' (SEQ ID NO: 712); (e) an antisense strand consisting of the nucleotide sequence 5'-aCfaaaAfgCfaAfaacAfgGfuCfusAfsg-3' (SEQ ID NO: 1269) and a sense strand consisting of the nucleotide sequence 5'-AfgAfcCfuGfUfUfuUfgCfuUfUfUfgUf-3' (SEQ ID NO: 718); (f) an antisense strand consisting of the nucleotide sequence 5'-asCfsaAfaAfgCfaAfaacAfgGfuCfuAfgsasa-3' (SEQ ID NO: 1369) and a sense strand consisting of the nucleotide sequence 5'-CfsusAfgAfcCfuGfUfUfuUfgCfuUfuUfgUf-3' (SEQ ID NO: 818); (g) an antisense strand consisting of the nucleotide sequence 5'-asCfsaAfaagCfaAfaacAfgGfucuAfgsasa-3' (SEQ ID NO:1660) and a sense strand consisting of the nucleotide sequence 5'-CfsusAfgAfcCfuGfUfUfuUfgcuuuugu-3' SEQ ID NO:1654); or (h) an antisense strand consisting of the nucleotide sequence 5'-asCfsaAfaAfsgCfaAfaacAfgGfuCfsuAfgsasa-3' (SEQ ID NO: 1400) and a sense strand consisting of the nucleotide sequence 5'-CfsusAfgAfcCfuGfUfUfuUfgCfsuUfsuUfsgsUfs-3' (SEQ ID NO: 849); wherein a, g, c, and u are 2'-O-methyl (2'-OMe) modified A, G, C, and U nucleotides, respectively; Af, Gf, Cf and Uf are 2' fluoro A, G, C and U modified nucleotides, respectively; dT is a 2'-deoxythymidine nucleotide and s is a phosphorothioate linkage. 23. A pharmaceutical composition comprising the double stranded RNAi agent of claim 1. 24. The pharmaceutical composition of claim 23, further 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, further comprising a buffer solution. 27. The pharmaceutical composition of claim 26, wherein the buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof. 28. A double stranded RNAi agent that inhibits the expression of Proprotein convertase subtilisin kexin 9 (PCSK9) in a cell, wherein said double stranded RNAi agent comprises a sense strand complementary to an antisense strand forming a double stranded region, wherein said antisense strand comprises a region complementary to part of an mRNA encoding PCSK9, wherein each strand is independently about 14 to about 30 nucleotides in length, wherein said antisense strand comprises at least 19 contiguous nucleotides of the nucleotide sequence 5'-ACAAAAGCAAAACAGGUCUAG-3' (SEQ ID NO: 412) and said double stranded RNAi agent is represented by formula Ma: TABLE-US-00038 sense: 5' n.sub.p-N.sub.a-Y Y Y-N.sub.a-n.sub.q 3' antisense: 3' n.sub.p'-N.sub.a'-Y'Y'Y'-N.sub.a'-n.sub.q' 5' (IIIa) wherein n.sub.p, n.sub.q, n.sub.p' and n.sub.q', each of which may or may not be present, each independently represents an overhang nucleotide; p, q, and q' are each independently 0-6; n.sub.p'>0 and at least one n.sub.p' is linked to a neighboring nucleotide via a phosphorothioate linkage; each N.sub.a and each N.sub.a' independently represents an oligonucleotide sequence comprising 0-25 nucleotides, 2-20 of which are modified nucleotides, wherein the modified nucleotides are each independently selected from the group consisting of 2'-O-methyl, 2'-fluoro, and 2'-deoxythymidine (dT); YYY and Y'Y'Y' each independently represent one motif of three identical modifications on three consecutive nucleotides, and wherein the modifications are 2'-O-methyl or 2'-fluoro modifications; wherein YYY is complementary to Y'Y'Y'; wherein the sense strand comprises at least one phosphorothioate linkage; and wherein the sense strand is conjugated to at least one ligand, wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker. 29. The double stranded RNAi agent of claim 28, wherein the YYY motif occurs at or near the cleavage site of the sense strand. 30. The double stranded RNAi agent of claim 1 or 28, wherein each N.sub.a and each N.sub.a' independently represents an oligonucleotide sequence comprising 0-25 nucleotides which are modified, each sequence comprising at least two differently modified nucleotides. 31. The double stranded RNAi agent of claim 1 or 28, wherein each N.sub.a and each N.sub.a' independently represents an oligonucleotide sequence comprising 0-25 nucleotides which are modified, each sequence comprising at least two differently modified nucleotides, wherein the modifications on the nucleotides are independently selected from the group consisting of 2'-O-methyl, 2'-fluoro, and 2'-deoxythymidine (dT). 32. The double stranded RNAi agent of claim 1 or 28, wherein each N.sub.b and each N.sub.b' independently represents an oligonucleotide sequence comprising 0-10 nucleotides which are modified. 33. The double stranded RNAi agent of claim 1 or 28, wherein each N.sub.b and each N.sub.b' independently represents an oligonucleotide sequence comprising 0-10 nucleotides which are modified, wherein the modifications on the nucleotides are independently selected from the group consisting of 2'-O-methyl, 2'-fluoro, and 2'-deoxythymidine (dT). 34. The double stranded RNAi agent of claim 1 or 28, wherein substantially all of the nucleotides of the sense strand and substantially all of the nucleotides of the antisense strand are modified nucleotides. 35. The double stranded RNAi agent of claim 1 or 28, wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand are modified. 36. The double stranded RNAi agent of claim 1 or 28, wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand are modified, and wherein the modifications on the nucleotides are independently selected from the group consisting of 2'-O-methyl, 2'-fluoro, and 2'-deoxythymidine (dT). 37. The double stranded RNAi agent of any one of claims 1, 2, 6, 9, 28, and 10-12, wherein the Y nucleotides contain a 2'-O-methyl modification and the Y' nucleotides contain a 2'-fluoro modification. 38. The double stranded RNAi agent of claim 1 or 28, wherein each strand is about 19 to about 25 nucleotides in length. 39. The double stranded RNAi agent of claim 1 or 28, wherein each strand is about 19 to about 23 nucleotides in length. 40. The double stranded RNAi agent of claim 1 or 28, wherein each strand is about 17 to about 23 nucleotides in length. 41. The double stranded RNAi agent of claim 1 or 28, wherein each strand is about 21 to about 25 nucleotides in length. 42. The double stranded RNAi agent of claim 1 or 28, wherein the ligand is ##STR00016## 43. The double stranded RNAi agent of claim 1 or 28, wherein the ligand is attached to the 3' end of the sense strand. 44. The double stranded RNAi agent of claim 43, wherein the RNAi agent is conjugated to the ligand as shown in the following schematic ##STR00017## wherein X is O or S. 45. A pharmaceutical composition comprising the double stranded RNAi agent of claim 28. 46. A double stranded RNAi agent that inhibits the expression of Proprotein convertase subtilisin kexin 9 (PCSK9) in a cell, wherein said double stranded RNAi agent comprises a sense strand complementary to an antisense strand forming a double stranded region, wherein said antisense strand comprises a region complementary to part of an mRNA encoding PCSK9, wherein each strand is independently about 14 to about 30 nucleotides in length, wherein said antisense strand comprises at least 19 contiguous nucleotides of the nucleotide sequence 5'-ACAAAAGCAAAACAGGUCUAG-3' (SEQ ID NO: 412), wherein said double stranded RNAi agent comprises at least one modified nucleotide selected from the group consisting of 2'-O-methyl, 2'-fluoro, and 2'-deoxythymidine (dT); wherein the antisense strand comprises two phosphorothioate internucleotide linkages between the 3'-end three terminal nucleotides and two phosphorothioate internucleotide linkages between the 5' end three terminal nucleotides; wherein the sense strand comprises two phosphorothioate internucleotide linkages between the 5' end three terminal nucleotides; and wherein the sense strand is conjugated to a ligand comprising one or more GalNAc derivatives attached through a bivalent or trivalent branched linker. 47. The dsRNA agent of claim 46, wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand are modified nucleotides. 48. The double stranded RNAi agent of claim 46, wherein the ligand is ##STR00018## 49. The double stranded RNAi agent of claim 46, wherein the ligand is attached to the 3' end of the sense strand. 50. The double stranded RNAi agent of claim 46, wherein the RNAi agent is conjugated to the ligand as shown in the following schematic ##STR00019## wherein X is O or S. 51. A pharmaceutical composition comprising the double stranded RNAi agent of claim 46. 52. A double stranded RNAi agent that inhibits the expression of Proprotein convertase subtilisin kexin 9 (PCSK9) in a cell, comprising a sense strand and an antisense strand, wherein the sense strand comprises the nucleotide sequence of 5'-csusagacCfuGfudTuugcuuuuguL96-3' (SEQ ID NO:1657) and the antisense strand comprises the nucleotide sequence of 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3' (SEQ ID NO:1663), 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; dT is 2'-deoxythymidine; and L96 is N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol. 53. An isolated cell containing the double stranded RNAi agent of claim 52. 54. A pharmaceutical composition comprising the double stranded RNAi agent of claim 52. 55. The pharmaceutical composition of claim 54, further comprising an unbuffered solution. 56. The pharmaceutical composition of claim 55, wherein the unbuffered solution is saline or water. 57. The pharmaceutical composition of claim 54, further comprising a buffer solution. 58. The pharmaceutical composition of claim 57, wherein the buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof. 59. A double stranded RNAi agent that inhibits the expression of Proprotein convertase subtilisin kexin 9 (PCSK9) in a cell, comprising a sense strand and an antisense strand, wherein the sense strand consists of the nucleotide sequence of 5'-csusagacCfuGfudTuugcuuuuguL96-3' (SEQ ID NO:1657) and the antisense strand consists of the nucleotide sequence of 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3' (SEQ ID NO:1663), 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; dT is 2'-deoxythymidine; and L96 is N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol. 60. A pharmaceutical composition comprising the double stranded RNAi agent of claim 59. 61. A double stranded RNAi agent that inhibits the expression of Proprotein convertase subtilisin kexin 9 (PCSK9) in a cell, comprising a sense strand which differs by no more than 4 bases from the nucleotide sequence of 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3' (SEQ ID NO:1663), and an antisense strand which differs by no more than 4 bases from the nucleotide sequence of 5'-csusagacCfuGfudTuugcuuuuguL96-3' (SEQ ID NO:1657), 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; dT is 2'-deoxythymidine; and L96 is N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol. 62. The double stranded RNAi agent of claim 61, wherein the sense strand differs by no more than 3 bases from the nucleotide sequence of 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3' (SEQ ID NO:1663), and the antisense strand differs by no more than 3 bases from the nucleotide sequence of 5'-csusagacCfuGfudTuugcuuuuguL96-3' (SEQ ID NO:1657). 63. The double stranded RNAi agent of claim 61, wherein the sense strand differs by no more than 2 bases from the nucleotide sequence of 5'-asCfsaAfAfAfgCfaAfaAfcAfgGfuCfuagsasa-3' (SEQ ID NO:1663), and the antisense strand differs by no more than 2 bases from the nucleotide sequence of 5'-csusagacCfuGfudTuugcuuuuguL96-3' (SEQ ID NO:1657). 64. An isolated cell containing the double stranded RNAi agent of any one of claims 1, 28, 46, 59, and 61. 65. A pharmaceutical composition comprising the double stranded RNAi agent of claim 61. 66. A method of inhibiting PCSK9 expression in a cell, the method comprising: (a) contacting the cell with the double stranded RNAi agent of any one of claim, 1, 28, 17-16, 96, 10-14, 20, 59 or 61 or a pharmaceutical composition of any one of claim 23, 45, 51, 60 or 40; and (b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a PCSK9 gene, thereby inhibiting expression of the PCSK9 gene in the cell. 67. A method of treating a subject having lipidemia mediated by PCSK9 expression, comprising administering to the subject a therapeutically effective amount of the double stranded RNAi agent of any one of claim, 1, 28, 17-16, 46, 10-14, 20, 59 or 61 or a pharmaceutical composition of any one of claim 23, 45, 51, 60 or 40, thereby treating said subject. 68. The method of claim 67, wherein the subject is a human. 69. The method of claim 68, wherein the human has hypercholesterolemia. 70. The method of claim 67, wherein the double stranded RNAi agent is administered at a dose of about 0.01 mg/kg to about 10 mg/kg or about 0.5 mg/kg to about 50 mg/kg. 71. The method of claim 67, wherein the double stranded RNAi agent is administered in two or more doses. 72. The method of claim 71, wherein the double stranded RNAi agent is administered in a dosing regimen that includes a loading phase followed by a maintenance phase. 73. The method of claim 72, wherein the maintenance phase comprises administering a dose of the double stranded RNAi agent to the subject once every three months. 74. The method of claim 72, wherein the maintenance phase comprises administering a dose of the double stranded RNAi agent to the subject once every six months. 75. The method of claim 67, wherein the double stranded RNAi agent or the pharmaceutical composition is administered to the subject subcutaneously or intravenously. 76. A method of inhibiting PCSK9 expression in a cell, the method comprising: (a) contacting the cell with the double stranded RNAi agent of claim 52 or the pharmaceutical composition of claim 54; and (b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a PCSK9 gene, thereby inhibiting expression of the PCSK9 gene in the cell. 77. A method of treating a subject having lipidemia mediated by PCSK9 expression, comprising administering to the subject a therapeutically effective amount of the double stranded RNAi agent of claim 52 or the pharmaceutical composition of claim 54, thereby treating said subject. 78. The method of claim 77, wherein the subject is a human. 79. The method of claim 78, wherein the human has hypercholesterolemia. 80. The method of claim 77, wherein the double stranded RNAi agent or the pharmaceutical composition is administered to the subject subcutaneously or intravenously. 81. The method of claim 77, wherein the double stranded RNAi agent is administered at a dose of about 0.01 mg/kg to about 10 mg/kg or about 0.5 mg/kg to about 50 mg/kg. 82. The method of claim 77, wherein the double stranded RNAi agent is administered in two or more doses. 83. The method of claim 77, wherein the double stranded RNAi agent is administered in a dosing regimen that includes a loading phase followed by a maintenance phase. 84. The method of claim 83, wherein the maintenance phase comprises administering a dose of the double stranded RNAi agent to the subject once every three months. 85. The method of claim 83, wherein the maintenance phase comprises administering a dose of the double stranded RNAi agent to the subject once every six months.