17β-HYDROXYSTEROID DEHYDROGENASE TYPE 13 (HSD17B13) iRNA COMPOSITIONS AND METHODS OF USE THEREOF

Abstract

To provide 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) iRNA compositions and methods of use thereof.SOLUTION: The present invention relates to double-stranded ribonucleic acid (dsRNA) compositions targeting the HSD17B13 gene, as well as methods of inhibiting expression of HSD17B13, and methods of treating subjects that would benefit from reduction in expression of HSD17B13, such as subjects having a HSD17B13-associated disease, disorder, or condition, using such dsRNA compositions. The present invention also provides methods of using the iRNA compositions of the invention for inhibiting the expression of a HSD17B13 gene and / or for treating a subject who would benefit from inhibiting or reducing the expression of a HSD17B13 gene, e.g., a subject suffering or prone to suffering from an HSD17B13-associated disease, for example, a chronic fibro-inflammatory liver disease.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of priority to U.S. Provisional Application No. 62 / 645,941, filed March 21, 2018, U.S. Provisional Application No. 62 / 770,298, filed November 21, 2018, and U.S. Provisional Application No. 62 / 775,590, filed December 5, 2018. The entire contents of each of the foregoing provisional patent applications are incorporated herein by reference. Sequence Listing

[0002] This application has been submitted electronically in ASCII format and contains a Sequence Listing, which is incorporated herein by reference in its entirety. The ASCII copy was created on March 12, 2019, is named 121301-08420_SL.txt, and is 1,374,047 bytes in size. [Background technology]

[0003] 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) is a member of the 17β-hydroxysteroid dehydrogenase (HSD17B) family of enzymes, which have a variety of functions, including the reduction or oxidation of sex hormones, fatty acids, and bile acids in vivo (Moeller and Adamski (2009) Mol Cell Endocrinol 301: 7). Members of the HSD17B family are diverse in tissue distribution, They have diverse substrate specificities, differing in subcellular localization, catalytic preferences, and ability to catalyze the conversion of substrates other than steroids, such as lipids and retinoids (Marchais-Oberwinkler, et al. (2011) J Steroid Biochem Mol Biol 125 (1-2): 66-82). HSD17B13 has been demonstrated to enhance hepatic lipogenesis in normal mouse liver and cultured human hepatocytes (Su, et al. (2014) Proc Natl Acad Sci USA 111: 11437).

[0004] Hepatocytes form the liver parenchyma and are involved in mobilizing lipids for energy and storing excess lipids in the form of lipid droplets (LDs), making the liver the major organ involved in lipid homeostasis.

[0005] LDs are now recognized as bioactive organelles involved in lipid metabolism, membrane trafficking, and signal transduction. LDs are generally composed of a core of neutral lipids (e.g., triacylglycerol (TG) and cholesterol esters surrounded by a phospholipid / cholesterol monolayer). Numerous LD-specific proteins are associated with LD membranes and functions, such as regulating the influx and efflux of molecules into and from LDs. The predominant hepatocyte LD-associated proteins are members of the perilipin family of proteins, but other proteins include hypoxia-inducible protein 2 (HIG2), patatin-like phospholipase domain-containing 3 (PNPLA3), and HSD17B13. Non-perilipin proteins have also been identified as LD-associated proteins (Carr and Ahima (2016) Exp Cell Res 15: 187; Su, et al. (2014) Proc Natl Acad Sci USA 111: 11437).

[0006] Increased accumulation of LDs is associated with many metabolic diseases and chronic fibroinflammatory liver diseases, such as hepatic fibrosis, NASH, and NAFLD. HSD17B13 has been identified as one of the most abundantly expressed LD proteins, specifically localized on the surface of LDs in human subjects and mice with NAFLD. It has also been shown that the expression level of HSD17B13 is upregulated in the livers of patients and mice with NAFLD. Overexpression of HSD17B13 results in an increase in the number and size of LDs. Overexpression of HSD17B13 in the liver of C57BL / 6 mice significantly increases hepatic lipogenesis and TG content, thereby leading to a fatty liver phenotype. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Moeller and Adamski (2009) Mol Cell Endocrinol 301: 7 [Non-patent document 2] Marchais-Oberwinkler, et al. (2011) J Steroid Biochem Mol Biol 125 (1-2): 66-82 [Non-patent document 3] Su, et al. (2014) Proc Natl Acad Sci USA 111: 11437 [Non-patent document 4] Carr and Ahima (2016) Exp Cell Res 15: 187;Su, et al. (2014) Proc Natl Acad Sci USA 111: 11437 Summary of the Invention [Means for solving the problem]

[0008] There is currently no treatment for chronic fibroinflammatory liver disease.The current standard treatment for the subject with chronic fibroinflammatory liver disease includes modifying lifestyle and managing related comorbidities, such as hypertension, hyperlipidemia, diabetes, obesity, etc.Therefore, the prevalence of chronic fibroinflammatory liver disease has been gradually increasing over the past decade and is expected to continue to increase, so there is a need in the art for alternative treatment for the subject with chronic fibroinflammatory liver disease.

[0009] The present invention provides an iRNA composition that induces RNA-induced silencing complex (RISC)-mediated cleavage of the RNA transcript of the 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) gene. The HSD17B13 gene can be present in a cell, for example, a cell in a subject, such as a human. The present invention also provides a method for using the iRNA composition of the present invention to inhibit the expression of the HSD17B13 gene and / or to treat a subject who is expected to benefit from inhibiting or reducing the expression of the HSD17B13 gene, for example, a subject who is suffering from or susceptible to an HSD17B13-related disease, such as chronic fibroinflammatory liver disease.

[0010] Thus, in one aspect, the present invention provides double-stranded ribonucleic acid (dsRNA) agents for inhibiting expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell. The dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from the nucleotide sequence of SEQ ID NO: 1 or 2, and the antisense strand comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from the nucleotide sequence of SEQ ID NO: 8 or 9. In some embodiments, the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises at least 15 contiguous nucleotides from the nucleotide sequence of SEQ ID NO: 1 or 2, and the antisense strand comprises at least 15 contiguous nucleotides from the nucleotide sequence of SEQ ID NO: 8 or 9.

[0011] In another aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell. The dsRNA agent comprises a sense strand and an antisense strand forming a double-stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding HSD17B13 comprising at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from any one of the antisense sequences listed in any one of Tables 2, 3, 7, 8, 10, 11, or 13. In some embodiments, the dsRNA agent comprises a sense strand and an antisense strand forming a double-stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding HSD17B13 comprising at least 15 contiguous nucleotides from any one of the antisense sequences listed in any one of Tables 2, 3, 7, 8, 10, 11, or 13.

[0012] In one embodiment, the region of complementarity is between nucleotides 213-242; 256-287; 361-385; 447-480; 483-529; 489-529; 630-653; 688-711; 752-777; 753-779; 772-806; 781-806; 791-851; 829-858; 870-896; 893-930; 900-930; 910-932; 980-1092; 1101-1102 of SEQ ID NO: 1. 158; 1176-1210; 1320-1350; 1335-1373; 1456-1482; 1506-1535; 1558-1588; 1699-1740; 1725-1757; 2182-2210; 2190-2254; 2194-2216; 2240-2373; or 2242-2264, containing at least 15 consecutive nucleotides that differ by no more than 1, 2, or 3 nucleotides. In some embodiments, the region of complementarity is between nucleotides 213-242; 256-287; 361-385; 447-480; 483-529; 489-529; 630-653; 688-711; 752-777; 753-779; 772-806; 781-806; 791-851; 829-858; 870-896; 893-930; 900-930; 910-932; 980-1000; 990-11000; 1010-11000; 1020-11000; 1030-11000; 1040-11000; 1050-11000; 1060-11000; 1070-11000; 1080-11000; 1090-11000; 1110-11100; 1120-11200; 1130-11300; 1140-11400; 1150-11500; 1160-11600; 1170-11700; 1180-11800; 1190-11900; 1210-12100; 1220-12200; 1230-12300; 1240-12400; 1250-12500; 1260-12600; 1270-12700; 1280-12800; 1 Contains at least 15 consecutive nucleotides from any one of: 092; 1101-1158; 1176-1210; 1320-1350; 1335-1373; 1456-1482; 1506-1535; 1558-1588; 1699-1740; 1725-1757; 2182-2210; 2190-2254; 2194-2216; 2240-2373; or 2242-2264.

[0013] In another embodiment, the region of complementarity is nucleotides 71-93, 101-123, 108-130, 109-131, 112-134, 123-145; 213-235; 220-242, 256-278; 292-314, 293-315, 299-321, 301-323, 324-346, 349-371, 350-372, 351-373, 352-374; 353-375, 355-377, 356-378, 357-379, 361-383; 363-385, 365-387, 402-424, 422-444, 423-445, 424-446, 425-447, 426-448, 427-449, 430-431, 431-432, 432-433, 433-434, 434-435, 435-436, 436-437, 437-438, 438-439, 439-440, 439-441, 439-442, 439-443, 439-444, 440-445, 441-442, 441-443, 441-445, 442-444, 442-445, 443-445, 444-445, 445-446, 445- 427-449, 428-450, 431-453, 447-469, 489-511, 490-512, 507-529, 541-563, 547-569, 548-570, 585-607, 589-611, 592-614, 593-615, 620-642, 630-646 52, 631-653, 632-654, 649-671, 676-698, 688-710, 723-745, 728-750, 752-774, 753-775, 755-777, 757-779, 763-785, 764-786, 772-794, 778-800, 78 0~802, 781~803, 791~813, 792~814, 794~816, 795~817, 807~829, 828~850, 829~851; 832~854; 836~858; 838~860; 839~861, 840~862, 832~861; 870~892 ;874~896, 894~916;895~917;896~918;897~919;898~920;899~921;900~922;901~923;902~924;906~928;908~930;894~930;910~932;965~987;966~ 988981~1003;1005~1027;1006~1028;1010~1032;1005~1032;1052~1074;1097~1119;1101~1123;1102~1124;1103~1125;1133~1155;1135~1157;113 6~1158;1097~1125;1133~1158;1176~1198;1188~1210;1243~1265;1315~13371320~1342;1322~1344;1325~1347;1327~1349;1328~1350;1320~1507;1335~1357;1336~1358;1458~1480;1459~1481;1460~1482;1458~1482;1497~1519;1498~1520;1506~1528;1513~1535;1565~1587;1566~1588;1613~1635;1614~1636;1622~1644;1643~1665;1699~1721;1717~1739;1718~1740 ;1724~1746;1725~1747;1726~1748;1727~1749;1728~1750;1717~1750;1737~1759;1768~1790;2188~2210;2190~2212;2188~2212;2194~2216;2195~2217;2250~2272;2232~2254;2240~2262;2232~2262;2242~2264;2245~2267 or 2347-2373; or 2347-2373; comprising at least 15 consecutive nucleotides that differ by no more than 1, 2, or 3 nucleotides from any one of 2249-2271; 2232-2271; 2347-2369; 2351-2373; or 2347-2373. In some embodiments, the region of complementarity is nucleotides 71-93, 101-123, 108-130, 109-131, 112-134, 123-145; 213-235; 220-242, 256-278; 292-314, 293-315, 299-321, 301-323, 324-346, 349-371, 350-372, 351-373, 352-374; 353-375, 355-377, 356-378, 357-379, 361-383 of SEQ ID NO:1;363~385、365~387、402~424、422~444、423~445、427~449、428~450、431~453、447~469、489~511、490~512、507~529、541~563、547~569、548~570、585~607、589~611、592~614、593~615、620~642、630~652、631~653、632~654、649~671、676~698、688~710、723~745、728~750、752~774、753~775、755~777、757~779、763~785、764~786、772~794、778~800、780~802、781~803、791~813、792~814、794~816、795~817、807~829、828~850、829~851;832~854;836~858;838~860;839~861、840~862、832~861;870~892;874~896、894~916;895~917;896~918;897~919;898~920;899~921;900~922;901~923;902~924;906~928;908~930;894~930;910~932;965~987;966~988981~1003;1005~1027;1006~1028;1010~1032;1005~1032;1052~1074;1097~1119;1101~1123;1102~1124;1103~1125;1133~1155;1135~1157;1136~1158;1097~1125;1133~1158;1176~1198;1188~1210;1243~1265;1315~13371320~1342;1322~1344;1325~1347;1327~1349;1328~1350;1320~1507;1335~1357;1336~1358;1458~1480;1459~1481;1460~1482;1458~1482;1497~1519;1498~1520;1506~1528;1513~1535;1565~1587;1566~1588;1613~1635;1614~1636;1622~1644;1643~1665;1699~1721;1717~1739;1718~1740;1724~1746;1725~1747;1726-1748; 1727-1749; 1728-1750; 1717-1750; 1737-1759; 1768-1790; 2188-2210; 2190-2212; 2188-2212; 2194-2216; 2195-2217; 2250-2272; 2232-2254; 2240-2262; 2232-2262; 2242-2264; 2245-2267 2249-2271; 2232-2271; 2347-2369; 2351-2373; or 2347-2373.

[0014] In another embodiment, the region of complementarity is between nucleotides 108-130; 109-131; 108-131; 112-134; 293-315; 301-323; 293-323; 361-383; 402-424; 423-445; 428-450; 423-450; 428-453; 431-453; 489-511; 490-512; 489-512; 649-671; 753-775; 772-794; 791-813; 792-814; 795-817; 791-818; 792-819; 793-819; 794-819; 795-819; 795-819; 791-819; 792-819; 793-819; 794 ...3-819; 794-819; 794-819; 795-819; 791-819; 791-819; 792-819; 792-819; 793-819; 793-819; 793-819; 793-819; 793-819; 7 7; 829-851; 832-854; 836-858; 829-858; 870-892; 874-896; 870-896; 898-920; 900-922; 902-924; 906-928; 908-930; 902-930; 910-932; 966-988; 1328-1350; or 2194-2216; 2242-2264; or 2249-2271, containing at least 15 consecutive nucleotides that differ by no more than 1, 2, or 3 nucleotides from any one of In some embodiments, the region of complementarity is between nucleotides 108-130; 109-131; 108-131; 112-134; 293-315; 301-323; 293-323; 361-383; 402-424; 423-445; 428-450; 423-450; 428-453; 431-453; 489-511; 490-512; 489-512; 649-671; 753-775; 772-794; 791-813; 792-814; 795- or 2249 to 2271. In another embodiment, the region of complementarity comprises at least 15 contiguous nucleotides from any one of nucleotides 817; 791-817; 829-851; 832-854; 836-858; 829-858; 870-892; 874-896; 870-896; 898-920; 900-922; 902-924; 906-928; 908-930; 902-930; 910-932; 966-988; 1328-1350; or 2194-2216; 2242-2264; or 2249-2271. In another embodiment, the region of complementarity comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from nucleotides 898-930 of SEQ ID NO:1. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides from nucleotides 898 to 930 of SEQ ID NO:1.

[0015] In one embodiment, the region of complementarity comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from nucleotides 910-932 of SEQ ID NO: 1. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides from nucleotides 910-932 of SEQ ID NO: 1.

[0016] In one embodiment, the region of complementarity comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from nucleotides 2194-2216 of SEQ ID NO: 1. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides from nucleotides 2194-2216 of SEQ ID NO: 1.

[0017] In one embodiment, the region of complementarity comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from nucleotides 2242-2264 of SEQ ID NO: 1. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides from nucleotides 2242-2264 of SEQ ID NO: 1.

[0018] In one embodiment, the dsRNA agent comprises at least one modified nucleotide.

[0019] In one embodiment, substantially all of the nucleotides in the sense strand comprise a modification. In another embodiment, substantially all of the nucleotides in the antisense strand comprise a modification. In yet another embodiment, substantially all of the nucleotides in the sense strand and substantially all of the nucleotides in the antisense strand comprise a modification.

[0020] In one aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell. The dsRNA agent comprises a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from the nucleotide sequence of SEQ ID NO: 1 or 2, and the antisense strand comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from the nucleotide sequence of SEQ ID NO: 8 or 9, wherein substantially all of the nucleotides in the sense strand and substantially all of the nucleotides in the antisense strand are modified nucleotides, and the sense strand is conjugated to a ligand attached to its 3' end. In some embodiments, the dsRNA agent comprises a sense strand and an antisense strand that form a double-stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides from the nucleotide sequence of SEQ ID NO: 1 or 2 and the antisense strand comprises at least 15 contiguous nucleotides from the nucleotide sequence of SEQ ID NO: 8 or 9, wherein substantially all of the nucleotides in the sense strand and substantially all of the nucleotides in the antisense strand are modified nucleotides, and the sense strand is conjugated to a ligand attached to its 3' end.

[0021] In one embodiment, all of the nucleotides in the sense strand comprise a modification. In another embodiment, all of the nucleotides in the antisense strand comprise a modification. In yet another embodiment, all of the nucleotides in the sense strand and all of the nucleotides in the antisense strand comprise a modification.

[0022] In one embodiment, at least one of the modified nucleotides is selected from the group consisting of a deoxy-nucleotide, a 3'-terminal deoxy-thymine (dT) nucleotide, a 2'-O-methyl modified nucleotide, a 2'-fluoro modified nucleotide, a 2'-deoxy modified nucleotide, a locked nucleotide, an unlocked nucleotide, a conformationally restricted nucleotide, a constrained ethyl nucleotide, an abasic nucleotide, a 2'-amino modified nucleotide, a 2'-O-allyl modified nucleotide, a 2'-C-alkyl modified nucleotide, a 2'-hydroxyl modified nucleotide, a 2'-methoxyethyl modified nucleotide, a 2'-O-alkyl modified nucleotide, a morpholino nucleotide, a phosphoramidate, a nucleotide comprising a non-natural base, a tetrahydropyran-modified nucleotide, a 1,5-anhydrohexitol-modified nucleotide, a cyclohexenyl-modified nucleotide, a nucleotide comprising a phosphorothioate group, a nucleotide comprising a methylphosphonate group, a nucleotide comprising a 5'-phosphate, a nucleotide comprising a 5'-phosphate mimic, a glycol-modified nucleotide, and a 2-O-(N-methylacetamido)-modified nucleotide, and combinations thereof.

[0023] In one embodiment, the nucleotide modifications are 2'-O-methyl and / or 2'-fluoro modifications.

[0024] The region of complementarity can be at least 17 nucleotides in length; 19-30 nucleotides in length; 19-25 nucleotides in length; or 21-23 nucleotides in length.

[0025] Each strand can be 30 nucleotides in length or less, e.g., each strand is independently 19 to 30 nucleotides in length; each strand is independently 19 to 25 nucleotides in length; each strand is independently 21 to 23 nucleotides in length.

[0026] The dsRNA can include at least one strand that includes a 3' overhang of at least one nucleotide; or at least one strand that includes a 3' overhang of at least two nucleotides.

[0027] In some embodiments, the dsRNA agent further comprises a ligand.

[0028] In one embodiment, the ligand is conjugated to the 3' end of the sense strand of the dsRNA agent.

[0029] In one embodiment, the ligand is an N-acetylgalactosamine (GalNAc) derivative.

[0030] In one embodiment, the ligand is [ka] is.

[0031] In one embodiment, the dsRNA agent is a ligand shown in the following schematic diagram: [ka] and X is O or S.

[0032] In one embodiment, X is O.

[0033] In one embodiment, the region of complementarity comprises any one of the antisense sequences in any one of Tables 2, 3, 7, 8, 10, 11, or 13.

[0034] In one aspect, the invention provides a duplex for inhibiting expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, the dsRNA agent comprising a sense strand complementary to an antisense strand, the antisense strand comprising a region complementary to a portion of an mRNA encoding HSD17B13, each strand being about 14 to about 30 nucleotides in length, the dsRNA agent having a structure represented by formula (III): Sense:5'n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -Na -n q 3' Antisense: 3'n p '-N a '-(X'X'X') k -N b '-Y'Y'Y'-N b '-(Z'Z'Z') l -N a '-n q '5'(III) (In the formula, i, j, k, and l are each independently 0 or 1; p, p', q, and q' are each independently 0 to 6; each N a and N a ' represents an oligonucleotide sequence containing 0-25 nucleotides that are independently modified or unmodified, or a combination thereof, each sequence containing at least two differently modified nucleotides; each N b and N b ' represents an oligonucleotide sequence containing 0-10 nucleotides, independently, modified or unmodified, or a combination thereof; each n p , n p ',n q , and n q ' each may be present or absent and independently represent an overhanging nucleotide; XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent a motif with three identical modifications on three consecutive nucleotides; N b The modification to N is different from the modification to Y. b (The modifier for ' is different from the modifier for Y') is represented by The sense strand is conjugated to at least one ligand.

[0035] In one embodiment, i is 0; j is 0; i is 1; j is 1; i and j are both 0; or i and j are both 1. In another embodiment, k is 0; l is 0; k is 1; l is 1; k and l are both 0; or k and l are both 1.

[0036] In one embodiment, XXX is complementary to X'X'X', YYY is complementary to Y'Y'Y', and ZZZ is complementary to Z'Z'Z'.

[0037] In one embodiment, the YYY motif is present at or near the cleavage site on the sense strand, eg, the Y'Y'Y' motif is present at positions 11, 12, and 13 from the 5' end on the antisense strand.

[0038] In one embodiment, formula (III) is represented by formula (IIIa): Sense:5'n p -N a -YYY-N a -n q 3' Antisense: 3'n p’ -N a’ -Y'Y'Y'-N a’ -n q’ 5'(IIIa). In another embodiment, formula (III) is represented by formula (IIIb): Sense:5'n p -N a -YYY-N b -ZZZ-N a -n q 3' Antisense: 3'n p’ -N a’ -Y'Y'Y'-N b’ -Z'Z'Z'-N a’ -n q’ 5'(IIIb) (In the formula, each N b and N b ' independently represents an oligonucleotide sequence containing 1 to 5 modified nucleotides) is expressed by

[0039] In yet another embodiment, formula (III) is represented by formula (IIIc): Sense:5'n p -N a -XXX-N b -YYY-N a -n q 3' Antisense: 3'n p’ -N a’ -X'X'X'-N b’ -Y'Y'Y'-N a’ -n q’ 5'(IIIc) (In the formula, each N b and N b ' independently represents an oligonucleotide sequence containing 1 to 5 modified nucleotides) is expressed by

[0040] In another embodiment, formula (III) is represented by formula (IIId): Sense:5'n p -N a -XXX-N b -YYY-N b -ZZZ-N a -n q 3' Antisense: 3'n p’ -N a’ -X'X'X'-N b’ -Y'Y'Y'-N b’ -Z'Z'Z'-N a’ -n q’ 5'(IIId) (In the formula, each N b and N b ' independently represents an oligonucleotide sequence containing 1 to 5 modified nucleotides, and each N a and N a ' independently represent an oligonucleotide sequence containing 2 to 10 modified nucleotides) is expressed by

[0041] The region of complementarity can be at least 17 nucleotides in length; 19-30 nucleotides in length; 19-25 nucleotides in length; or 21-23 nucleotides in length.

[0042] Each strand can be 30 nucleotides or less in length, for example, each strand is independently 19-30 nucleotides in length.

[0043] In one embodiment, the modification to the nucleotide is selected from the group consisting of LNA, HNA, CeNA, 2'-methoxyethyl, 2'-O-alkyl, 2'-O-allyl, 2'-C-allyl, 2'-fluoro, 2'-O-methyl, 2'-deoxy, 2'-hydroxyl, and combinations thereof.

[0044] In one embodiment, the modification to the nucleotide is a 2'-O-methyl or a 2'-fluoro modification.

[0045] In one embodiment, Y' is a 2'-O-methyl or 2'-fluoro modified nucleotide.

[0046] In one embodiment, at least one strand of the dsRNA agent can include a 3' overhang of at least 1 nucleotide; or a 3' overhang of at least 2 nucleotides.

[0047] In one embodiment, the dsRNA agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage.

[0048] In one embodiment, the phosphorothioate or methylphosphonate internucleotide linkage is at the 3'-end of one strand. In one embodiment, the strand is the antisense strand. In another embodiment, the strand is the sense strand.

[0049] In one embodiment, the phosphorothioate or methylphosphonate internucleotide linkage is at the 5'-end of one strand. In one embodiment, the strand is the antisense strand. In another embodiment, the strand is the sense strand.

[0050] In one embodiment, the strand is the antisense strand, hi another embodiment, the strand is the sense strand.

[0051] In one embodiment, phosphorothioate or methylphosphonate internucleotide linkages are at both the 5' and 3' ends of one strand.

[0052] In one embodiment, the first base pair at the 5' end of the antisense strand of the duplex is an AU base pair.

[0053] In one embodiment, p'>0. In another embodiment, p'=2.

[0054] In one embodiment, q'=0, p=0, q=0, and the p' overhanging nucleotides are complementary to the target mRNA. In another embodiment, q'=0, p=0, q=0, and the p' overhanging nucleotides are non-complementary to the target mRNA.

[0055] In one embodiment, the sense strand has a total of 21 nucleotides and the antisense strand has a total of 23 nucleotides.

[0056] In one embodiment, at least one n p In another embodiment, all n' are linked to adjacent nucleotides by phosphorothioate linkages. p ' is linked to the adjacent nucleotide by a phosphorothioate linkage.

[0057] In one embodiment, all of the nucleotides in the sense strand and all of the nucleotides in the antisense strand comprise a modification.

[0058] In one embodiment, the ligand is conjugated to the 3' end of the sense strand of the dsRNA agent.

[0059] In one embodiment, the ligand is one or more N-acetylgalactosamine (GalNAc) derivatives attached through a monovalent, divalent, or trivalent branched linker.

[0060] In one embodiment, the ligand is [ka] is.

[0061] In one embodiment, the dsRNA agent is a ligand shown in the following schematic diagram: [ka] and X is O or S.

[0062] In one embodiment, X is O.

[0063] In one aspect, the invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell. The dsRNA agent comprises a sense strand complementary to an antisense strand, the antisense strand comprising a region complementary to a portion of an mRNA encoding HSD17B13, each strand being about 14 to about 30 nucleotides in length, and the dsRNA agent has the formula (III): Sense:5'n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3' Antisense: 3'n p '-N a '-(X'X'X') k -N b '-Y'Y'Y'-N b '-(Z'Z'Z') l -N a '-n q '5'(III) (In the formula, i, j, k, and l are each independently 0 or 1; p, p', q, and q' are each independently 0 to 6; each N a and N a ' represents an oligonucleotide sequence containing 0-25 nucleotides that are independently modified or unmodified, or a combination thereof, each sequence containing at least two differently modified nucleotides; each N b and N b ' represents an oligonucleotide sequence containing 0-10 nucleotides, independently, modified or unmodified, or a combination thereof; each n p , n p ',n q , and n q ' each may be present or absent and independently represent an overhanging nucleotide; XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent one motif with three identical modifications to three consecutive nucleotides, the modifications being 2'-O-methyl or 2'-fluoro modifications; N b The modification to N is different from the modification to Y. b (The modifier for ' is different from the modifier for Y') is represented by The sense strand is conjugated to at least one ligand.

[0064] In one aspect, the invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell. The dsRNA agent comprises a sense strand complementary to an antisense strand, the antisense strand comprising a region complementary to a portion of an mRNA encoding HSD17B13, each strand being about 14 to about 30 nucleotides in length, and the dsRNA agent has the formula (III): Sense:5'n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -nq 3' Antisense: 3'n p '-N a '-(X'X'X') k -N b '-Y'Y'Y'-N b '-(Z'Z'Z') l -N a '-n q '5'(III) (In the formula, i, j, k, and l are each independently 0 or 1; each n p , n q , and n q ' each may be present or absent and independently represent an overhanging nucleotide; p, q, and q' are each independently 0 to 6; n p '>0 and at least one n p ' is linked to adjacent nucleotides by phosphorothioate linkages; each N a and N a ' represents an oligonucleotide sequence containing 0-25 nucleotides that are independently modified or unmodified, or a combination thereof, each sequence containing at least two differently modified nucleotides; each N b and N b ' represents an oligonucleotide sequence containing 0-10 nucleotides, independently, modified or unmodified, or a combination thereof; XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent one motif with three identical modifications to three consecutive nucleotides, the modifications being 2'-O-methyl or 2'-fluoro modifications; N b The modification to N is different from the modification to Y. b (The modifier for ' is different from the modifier for Y') is represented by The sense strand is conjugated to at least one ligand.

[0065] In one aspect, the invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell. The dsRNA agent comprises a sense strand complementary to an antisense strand, the antisense strand comprising a region complementary to a portion of an mRNA encoding HSD17B13, each strand being about 14 to about 30 nucleotides in length, and the dsRNA agent has the formula (III): Sense:5'n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3' Antisense: 3'n p '-N a '-(X'X'X') k -N b '-Y'Y'Y'-N b '-(Z'Z'Z') l -N a '-n q '5'(III) (In the formula, i, j, k, and l are each independently 0 or 1; each n p , n q , and n q ' each may be present or absent and independently represent an overhanging nucleotide; p, q, and q' are each independently 0 to 6; n p '>0 and at least one n p ' is linked to adjacent nucleotides by phosphorothioate linkages; each N a and N a' represents an oligonucleotide sequence containing 0-25 nucleotides that are independently modified or unmodified, or a combination thereof, each sequence containing at least two differently modified nucleotides; each N b and N b ' represents an oligonucleotide sequence containing 0-10 nucleotides, independently, modified or unmodified, or a combination thereof; XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent one motif with three identical modifications to three consecutive nucleotides, the modifications being 2'-O-methyl or 2'-fluoro modifications; N b The modification to N is different from the modification to Y. b (The modifier for ' is different from the modifier for Y') is represented by The sense strand is conjugated to at least one ligand, which is one or more GalNAc derivatives attached through a monovalent, divalent, or trivalent branched linker.

[0066] In one aspect, the invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell. The dsRNA agent comprises a sense strand complementary to an antisense strand, the antisense strand comprising a region complementary to a portion of an mRNA encoding HSD17B13, each strand being about 14 to about 30 nucleotides in length, and the dsRNA agent has the formula (III): Sense:5'n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3' Antisense: 3'n p '-N a '-(X'X'X') k -Nb '-Y'Y'Y'-N b '-(Z'Z'Z') l -N a '-n q '5'(III) (In the formula, i, j, k, and l are each independently 0 or 1; each n p , n q , and n q ' each may be present or absent and independently represent an overhanging nucleotide; p, q, and q' are each independently 0 to 6; n p '>0 and at least one n p ' is linked to adjacent nucleotides by phosphorothioate linkages; each N a and N a ' represents an oligonucleotide sequence containing 0-25 nucleotides that are independently modified or unmodified, or a combination thereof, each sequence containing at least two differently modified nucleotides; each N b and N b ' represents an oligonucleotide sequence containing 0-10 nucleotides, independently, modified or unmodified, or a combination thereof; XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent one motif with three identical modifications to three consecutive nucleotides, the modifications being 2'-O-methyl or 2'-fluoro modifications; N b The modification to N is different from the modification to Y. b (The modifier for ' is different from the modifier for Y') is represented by the sense strand comprises at least one phosphorothioate linkage; The sense strand is conjugated to at least one ligand, which is one or more GalNAc derivatives attached through a monovalent, divalent, or trivalent branched linker.

[0067] In one aspect, the invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell. The dsRNA agent comprises a sense strand complementary to an antisense strand, the antisense strand comprising a region complementary to a portion of an mRNA encoding HSD17B13, each strand being about 14 to about 30 nucleotides in length, and the dsRNA agent has the formula (III): Sense:5'n p -N a -YYY-N a -n q 3' Antisense: 3'n p '-N a '-Y'Y'Y'-N a '-n q '5'(IIIa) (In the formula, each n p , n q , and n q ' each may be present or absent and independently represent an overhanging nucleotide; p, q, and q' are each independently 0 to 6; n p '>0 and at least one n p ' is linked to adjacent nucleotides by phosphorothioate linkages; each N a and N a ' represents an oligonucleotide sequence containing 0-25 nucleotides that are independently modified or unmodified, or a combination thereof, each sequence containing at least two differently modified nucleotides; YYY and Y'Y'Y' each independently represent one motif with three identical modifications to three consecutive nucleotides, the modifications being 2'-O-methyl and / or 2'-fluoro modifications. is represented by the sense strand comprises at least one phosphorothioate linkage; The sense strand is conjugated to at least one ligand, which is one or more GalNAc derivatives attached through a monovalent, divalent, or trivalent branched linker.

[0068] In one aspect, the invention provides double-stranded ribonucleic acid (dsRNA) agents for inhibiting the expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell. The dsRNA agent comprises a sense strand and an antisense strand that form a double-stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from the nucleotide sequence of SEQ ID NO: 1 or 2, and the antisense strand comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from the nucleotide sequence of SEQ ID NO: 8 or 9, wherein substantially all of the nucleotides in the sense strand comprise a modification selected from the group consisting of a 2'-O-methyl modification and a 2'-fluoro modification, wherein the sense strand comprises two phosphorothioate internucleotide linkages at the 5'-terminus and substantially all of the nucleotides in the antisense strand comprise a modification selected from the group consisting of a 2'-O-methyl modification and a 2'-fluoro modification, wherein the antisense strand comprises two phosphorothioate internucleotide linkages at the 5'-terminus and two phosphorothioate internucleotide linkages at the 3'-terminus, and wherein the sense strand is conjugated at its 3'-terminus to one or more GalNAc derivatives attached through a monovalent, divalent, or trivalent branched linker. In some embodiments, a dsRNA agent comprises a sense strand and an antisense strand that form a double-stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides from the nucleotide sequence of SEQ ID NO: 1 or 2, and the antisense strand comprises at least 15 contiguous nucleotides from the nucleotide sequence of SEQ ID NO: 8 or 9, wherein substantially all of the nucleotides of the sense strand comprise a modification selected from the group consisting of a 2'-O-methyl modification and a 2'-fluoro modification, the sense strand comprises two phosphorothioate internucleotide linkages at the 5'-terminus, and substantially all of the nucleotides of the antisense strand comprise a modification selected from the group consisting of a 2'-O-methyl modification and a 2'-fluoro modification, the antisense strand comprises two phosphorothioate internucleotide linkages at the 5'-terminus and two phosphorothioate internucleotide linkages at the 3'-terminus, and the sense strand is conjugated at its 3'-terminus to one or more GalNAc derivatives attached through a monovalent, divalent, or trivalent branched linker.

[0069] In one embodiment, all of the nucleotides in the sense strand and all of the nucleotides in the antisense strand are modified nucleotides.

[0070] In one embodiment, the region of complementarity comprises any one of the antisense sequences listed in any one of Tables 2, 3, 7, 8, 10, 11, or 13. In one embodiment, the agent is selected from the group consisting of AD-288917, AD-288996, AD-413639, AD-413644, and AD-413669. In one embodiment, the RNAi agent is AD-288917. In another embodiment, the agent is AD-288996. In another embodiment, the agent is AD-413639. In one embodiment, the agent is AD-413644. In another embodiment, the agent is AD-413669

[0071] In one embodiment, the sense and antisense strands comprise a nucleotide sequence selected from the group consisting of the nucleotide sequences of any one of the agents listed in any one of Tables 2, 3, 7, 8, 10, 11, or 13.

[0072] The present invention also provides cells, vectors and pharmaceutical compositions that contain any of the dsRNA agents of the present invention.The dsRNA agent can be formulated in unbuffered solution, such as saline or water, or in buffered solution, such as acetate, citrate, prolamin, carbonate, or phosphate, or any combination thereof.In one embodiment, the buffered solution is phosphate buffered saline (PBS).

[0073] In one aspect, the present invention provides a method for inhibiting 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) expression in a cell, comprising contacting the cell with a dsRNA agent or pharmaceutical composition of the present invention, thereby inhibiting expression of HSD17B13 in the cell.

[0074] The cell can be in a subject, such as a human subject.

[0075] In one embodiment, HSD17B13 expression is inhibited by at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or is inhibited to below the level of detection of HSD17B13 expression.

[0076] In one embodiment, the human subject suffers from a disease, disorder, or condition associated with HSD17B13. In one embodiment, the disease, disorder, or condition associated with HSD17B13 is a chronic fibroinflammatory liver disease. In one embodiment, the chronic fibroinflammatory liver disease is selected from the group consisting of liver inflammation, liver fibrosis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cirrhosis, alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), HCV-related cirrhosis, drug-induced liver injury, and hepatocellular necrosis.

[0077] In one aspect, the present invention provides a method for inhibiting the expression of HSD17B13 in a subject, comprising administering to the subject a therapeutically effective amount of a dsRNA agent or pharmaceutical composition of the present invention, thereby inhibiting the expression of HSD17B13 in the subject.

[0078] In another aspect, the present invention provides a method for treating a subject suffering from HSD17B13-related disease, disorder or condition.The method comprises administering to the subject a therapeutically effective amount of the dsRNA agent or pharmaceutical composition of the present invention, thereby treating the subject suffering from HSD17B13-related disease, disorder or condition.

[0079] In another aspect, the present invention provides the method for preventing at least one symptom in the subject with the disease, disorder or condition that is expected to be beneficial by reducing the expression of HSD17B13 gene.Method comprises: administering to the subject a prophylactically effective amount of the dsRNA agent or pharmaceutical composition of the present invention, thereby preventing at least one symptom in the subject with the disease, disorder or condition that is expected to be beneficial by reducing the expression of HSD17B13 gene.

[0080] In another aspect, the present invention provides a method for reducing the risk of developing chronic liver disease in a subject with steatosis.The method comprises administering to the subject a therapeutically effective amount of the dsRNA agent or pharmaceutical composition of the present invention, thereby reducing the risk of developing chronic liver disease in the subject with steatosis.

[0081] In yet another aspect, the present invention provides a method for inhibiting the progression of steatosis to steatohepatitis in a subject suffering from steatosis.The method comprises administering to the subject a therapeutically effective amount of a dsRNA agent or pharmaceutical composition of the present invention, thereby inhibiting the progression of steatosis to steatohepatitis in the subject.

[0082] In one aspect, the present invention provides the method for inhibiting the accumulation of lipid droplets in the liver of the subject suffering from the disease, disorder or condition associated with HSD17B13.The method comprises administering to the subject a therapeutically effective amount of the dsRNA agent or pharmaceutical composition of the present invention and the dsRNA agent that targets PNPLA3 gene or the pharmaceutical composition that comprises the dsRNA agent that targets PNPLA3 gene, thereby inhibiting the accumulation of lipids in the liver of the subject suffering from the disease, disorder or condition associated with HSD17B13.

[0083] In another aspect, the present invention provides a method for treating a subject suffering from a disease, disorder or condition associated with HSD17B13.The method comprises administering to the subject a therapeutically effective amount of the dsRNA agent or pharmaceutical composition of the present invention and a dsRNA agent that targets the PNPLA3 gene or a pharmaceutical composition comprising a dsRNA agent that targets the PNPLA3 gene, thereby treating the subject suffering from a disease, disorder or condition associated with HSD17B13.

[0084] In another aspect, the present invention provides a method for preventing at least one symptom in a subject with a disease, disorder or condition that is expected to benefit from reducing the expression of the HSD17B13 gene.The method comprises administering to the subject a therapeutically effective amount of the dsRNA agent or pharmaceutical composition of the present invention and a dsRNA agent that targets the PNPLA3 gene or a pharmaceutical composition comprising a dsRNA agent that targets the PNPLA3 gene, thereby preventing at least one symptom in a subject with a disease, disorder or condition that is expected to benefit from reducing the expression of the HSD17B13 gene.

[0085] In another aspect, the present invention provides a method for reducing the risk of developing chronic liver disease in a subject with steatosis.The method comprises administering to the subject a therapeutically effective amount of the dsRNA agent or pharmaceutical composition of the present invention and a dsRNA agent that targets the PNPLA3 gene or a pharmaceutical composition that comprises a dsRNA agent that targets the PNPLA3 gene, thereby reducing the risk of developing chronic liver disease in a subject with steatosis.

[0086] In another aspect, the present invention provides a method for inhibiting the progression of steatosis to steatohepatitis in a subject suffering from steatosis.The method comprises administering to the subject a therapeutically effective amount of the dsRNA agent or pharmaceutical composition of the present invention and a dsRNA agent targeting the PNPLA3 gene or a pharmaceutical composition comprising a dsRNA agent targeting the PNPLA3 gene, thereby inhibiting the progression of steatosis to steatohepatitis in the subject.

[0087] In one embodiment, administration of the dsRNA agent or pharmaceutical composition to a subject results in a decrease in HSD17B13 enzyme activity, a decrease in HSD17B13 protein accumulation, a decrease in PNPLA3 enzyme activity, a decrease in PNPLA3 protein accumulation, and / or a reduction in fat accumulation and / or an increase in lipid droplets in the subject's liver.

[0088] In one embodiment, the HSD17B13-associated disease, disorder, or condition is chronic fibroinflammatory liver disease.

[0089] In one embodiment, the chronic fibroinflammatory liver disease is selected from the group consisting of fat accumulation in the liver, liver inflammation, liver fibrosis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cirrhosis, alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), HCV-associated cirrhosis, drug-induced liver injury, and hepatocellular necrosis.

[0090] In one embodiment, the chronic fibroinflammatory liver disease is non-alcoholic steatohepatitis (NASH).

[0091] In one embodiment, the subject is obese.

[0092] In one embodiment, the methods and uses of the invention further comprise administering to the subject an additional therapeutic agent.

[0093] In one embodiment, the dsRNA agent is administered to the subject 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.

[0094] The agent can be administered to the subject intravenously, intramuscularly, or subcutaneously, hi one embodiment, the agent is administered to the subject subcutaneously.

[0095] In one embodiment, the methods and uses of the present invention further comprise determining the level of HSD17B13 in the subject.

[0096] In one aspect, the invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, the dsRNA agent comprising a sense strand and an antisense strand that form a duplex region, wherein the sense strand comprises the nucleotide sequence of any one of the agents in any one of Tables 2, 3, 7, 8, 10, 11, or 13, and the antisense strand comprises the nucleotide sequence of any one of the agents in any one of Tables 2, 3, 7, 8, 10, 11, or 13, wherein substantially all of the nucleotides in the sense strand and substantially all of the nucleotides in the antisense strand are modified nucleotides; and the dsRNA agent is conjugated to a ligand. In an embodiment of the present invention, for example, the following items are provided: (Item 1) A double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, comprising a sense strand and an antisense strand, wherein the sense strand comprises at least 15 consecutive nucleotides that differ by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO: 1 or 2, and the antisense strand comprises at least 15 consecutive nucleotides that differ by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO: 8 or 9. (Item 2) 1. A double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, the dsRNA agent comprising a sense strand and an antisense strand that form a double-stranded region, the antisense strand comprising a region of complementarity to an mRNA encoding HSD17B13 that comprises at least 15 contiguous nucleotides that differ by no more than 3 nucleotides from any one of the antisense sequences listed in any one of Tables 2, 3, 7, 8, 10, 11, or 13. (Item 3) 3. The dsRNA agent of item 1 or 2, comprising at least one modified nucleotide. (Item 4) 4. The dsRNA agent of any one of items 1 to 3, wherein substantially all of the nucleotides of the sense strand comprise a modification. (Item 5) 4. The dsRNA agent of any one of items 1 to 3, wherein substantially all of the nucleotides of the antisense strand comprise a modification. (Item 6) 4. The dsRNA agent of any one of items 1 to 3, wherein substantially all of the nucleotides of the sense strand and substantially all of the nucleotides of the antisense strand comprise a modification. (Item 7) A double-stranded RNA (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, comprising a sense strand and an antisense strand forming a double-stranded region, the sense strand comprises at least 15 consecutive nucleotides that differ by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO: 1 or 2, and the antisense strand comprises at least 15 consecutive nucleotides that differ by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO: 8 or 9; substantially all of the nucleotides of the sense strand and substantially all of the nucleotides of the antisense strand are modified nucleotides; The dsRNA agent, wherein the sense strand is conjugated to a ligand attached to the 3' end. (Item 8) Item 9. The dsRNA agent according to Item 7, wherein all of the nucleotides of the sense strand contain a modification. 8. The dsRNA agent of item 7, wherein all of the nucleotides of the antisense strand contain a modification. (Item 10) 8. The dsRNA agent of item 7, wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a modification. (Item 11) At least one of the modified nucleotides is a deoxy-nucleotide, a 3'-terminal deoxy-thymine (dT) nucleotide, a 2'-O-methyl modified nucleotide, a 2'-fluoro modified nucleotide, a 2'-deoxy modified nucleotide, a locked nucleotide, an unlocked nucleotide, a conformationally restricted nucleotide, a constrained ethyl nucleotide, an abasic nucleotide, a 2'-amino modified nucleotide, a 2'-O-allyl modified nucleotide, a 2'-C-alkyl modified nucleotide, a 2'-hydroxyl modified nucleotide, a 2'-methoxyethyl modified nucleotide, a 2'-O-alkyl modified nucleotide, a 11. The dsRNA agent of any one of items 3 to 10, wherein the dsRNA agent is selected from the group consisting of nucleotides, morpholino nucleotides, phosphoramidates, nucleotides containing unnatural bases, tetrahydropyran-modified nucleotides, 1,5-anhydrohexitol-modified nucleotides, cyclohexenyl-modified nucleotides, nucleotides containing phosphorothioate groups, nucleotides containing methylphosphonate groups, nucleotides containing 5'-phosphates, nucleotides containing 5'-phosphate mimetics, glycol-modified nucleotides, and 2-O-(N-methylacetamido)-modified nucleotides, and combinations thereof. (Item 12) 12. The dsRNA agent of claim 11, wherein the nucleotide modification is a 2'-O-methyl and / or a 2'-fluoro modification. (Item 13) 13. The dsRNA agent of any one of items 1 to 12, wherein the region of complementarity is at least 17 nucleotides in length. (Item 14) 14. The dsRNA agent of any one of items 1 to 13, wherein the complementary region is 19 to 30 nucleotides in length. (Item 15) 15. The dsRNA agent according to item 14, wherein the complementary region is 19 to 25 nucleotides in length. (Item 16) 16. The dsRNA agent according to item 15, wherein the complementary region is 21 to 23 nucleotides in length. (Item 17) 17. The dsRNA agent of any one of paragraphs 1 to 16, wherein each strand is 30 nucleotides or less in length. (Item 18) 18. The dsRNA agent of any one of items 1 to 17, wherein each strand is independently 19 to 30 nucleotides in length. (Item 19) 19. The dsRNA agent of item 18, wherein each strand is independently 19 to 25 nucleotides in length. (Item 20) 19. The dsRNA agent of item 18, wherein each strand is independently 21 to 23 nucleotides in length. (Item 21) 21. The dsRNA agent of any one of paragraphs 1 to 20, wherein at least one strand comprises a 3' overhang of at least one nucleotide. (Item 22) 22. The dsRNA agent of claim 21, wherein at least one strand comprises a 3' overhang of at least 2 nucleotides. (Item 23) 23. The dsRNA agent of any one of items 1 to 6 and 11 to 22, further comprising a ligand. (Item 24) 24. The dsRNA agent of claim 23, wherein the ligand is conjugated to the 3' end of the sense strand of the dsRNA agent. (Item 25) 25. The dsRNA agent of item 7 or 24, wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative. (Item 26) The ligand is [ka] 26. The dsRNA agent according to Item 25, (Item 27) Schematic diagram below [ka] 27. The dsRNA agent of claim 26, wherein the dsRNA agent is conjugated to a ligand shown in (Item 28) 28. The dsRNA agent of item 27, wherein X is O. (Item 29) 3. The dsRNA agent of item 2, wherein the region of complementarity comprises any one of the antisense sequences in any one of Tables 2, 3, 7, 8, 10, 11, or 13. (Item 30) A double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, the dsRNA agent comprising a sense strand complementary to an antisense strand, the antisense strand comprising a region complementary to a portion of an mRNA encoding HSD17B13, and each strand being about 14 to about 30 nucleotides in length, the dsRNA agent having formula (III): Sense:5'n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3' antisense: 3'n p '-N a '-(X'X'X') k -N b '-Y'Y'Y'-N b '-(Z'Z'Z') l -N a '-n q '5'(III) (In the formula, i, j, k, and l are each independently 0 or 1; p, p', q, and q' are each independently 0 to 6; each N a and N a ' represents an oligonucleotide sequence containing 0-25 nucleotides that are independently modified or unmodified, or a combination thereof, each sequence containing at least two differently modified nucleotides; each Nb and N b ' represents an oligonucleotide sequence containing 0-10 nucleotides, independently, modified or unmodified, or a combination thereof; each n p , n p ',n q , and n q ' each may be present or absent and independently represent an overhanging nucleotide; XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent a motif with three identical modifications on three consecutive nucleotides; N b The modification to N is different from the modification to Y. b (The modifier for ' is different from the modifier for Y') is represented by The dsRNA agent, wherein the sense strand is conjugated to at least one ligand. (Item 31) 31. The dsRNA agent of item 30, wherein i is 0; j is 0; i is 1; j is 1; i and j are both 0; or i and j are both 1. (Item 32) 31. The dsRNA agent of item 30, wherein k is 0; l is 0; k is 1; l is 1; k and l are both 0; or k and l are both 1. (Item 33) 31. The dsRNA agent of item 30, wherein XXX is complementary to X'X'X', YYY is complementary to Y'Y'Y', and ZZZ is complementary to Z'Z'Z'. (Item 34) 31. The dsRNA agent of claim 30, wherein the YYY motif is present at or near the cleavage site of the sense strand. (Item 35) 31. The dsRNA agent according to item 30, wherein the Y'Y'Y' motif is present at positions 11, 12, and 13 from the 5' end of the antisense strand. (Item 36) Formula (III) may be replaced by formula (IIIa): Sense:5'n p -N a -YYY-N a -n q 3' Antisense: 3'n p’ -N a’ -Y'Y'Y'-N a’ -n q’ 31. The dsRNA agent according to item 30, represented by 5'(IIIa). (Item 37) Formula (III) may be replaced by formula (IIIb): Sense:5'n p -N a -YYY-N b -ZZZ-N a -n q 3' Antisense: 3'n p’ -N a’ -Y'Y'Y'-N b’ -Z'Z'Z'-N a’ -n q’ 5'(IIIb) (In the formula, each N b and N b ' independently represents an oligonucleotide sequence containing 1 to 5 modified nucleotides) 31. The dsRNA agent according to Item 30, represented by: (Item 38) Formula (III) may be replaced by formula (IIIc): Sense:5'n p -N a -XXX-N b -YYY-N a -n q 3' Antisense: 3'n p’ -N a’ -X'X'X'-N b’ -Y'Y'Y'-N a’ -n q’ 5'(IIIc) (In the formula, each N b and N b ' independently represents an oligonucleotide sequence containing 1 to 5 modified nucleotides) 31. The dsRNA agent according to item 30, wherein the dsRNA agent is represented by: (Item 39) Formula (III) may be replaced by formula (IIId): Sense:5'n p -N a -XXX-N b -YYY-N b -ZZZ-N a -n q 3' Antisense: 3'n p’ -N a’ -X'X'X'-N b’ -Y'Y'Y'-N b’ -Z'Z'Z'-N a’ -n q’ 5'(IIId) (In the formula, each N b and N b ' independently represents an oligonucleotide sequence containing 1 to 5 modified nucleotides, and each N a and N a ' independently represent an oligonucleotide sequence containing 2 to 10 modified nucleotides) 31. The dsRNA agent according to item 30, wherein the dsRNA agent is represented by: (Item 40) 40. The dsRNA agent of any one of items 30 to 39, wherein the region of complementarity is at least 17 nucleotides in length. (Item 41) 40. The dsRNA agent of any one of items 30 to 39, wherein the region of complementarity is 19 to 30 nucleotides in length. (Item 42) 42. The dsRNA agent according to item 41, wherein the complementary region is 19 to 25 nucleotides in length. (Item 43) 43. The dsRNA agent according to item 42, wherein the complementary region is 21 to 23 nucleotides in length. (Item 44) 44. The dsRNA agent of any one of paragraphs 30 to 43, wherein each strand is 30 nucleotides or less in length. (Item 45) 44. The dsRNA agent of any one of paragraphs 30 to 43, wherein each strand is independently 19 to 30 nucleotides in length. (Item 46) 46. ​​The agent of any one of items 30 to 45, wherein the modification on the nucleotide is selected from the group consisting of LNA, HNA, CeNA, 2'-methoxyethyl, 2'-O-alkyl, 2'-O-allyl, 2'-C-allyl, 2'-fluoro, 2'-O-methyl, 2'-deoxy, 2'-hydroxyl, and combinations thereof. (Item 47) 47. The dsRNA agent of claim 46, wherein the modification to the nucleotide is a 2'-O-methyl and / or a 2'-fluoro modification. (Item 48) 47. The dsRNA agent of any one of items 30 to 46, wherein Y' is a 2'-O-methyl or 2'-fluoro modified nucleotide. (Item 49) 49. The dsRNA agent of any one of paragraphs 30 to 48, wherein at least one strand comprises a 3' overhang of at least one nucleotide. (Item 50) 50. The dsRNA agent of any one of paragraphs 30 to 49, wherein at least one strand comprises a 3' overhang of at least 2 nucleotides. (Item 51) 51. The dsRNA agent of any one of paragraphs 30 to 50, wherein the dsRNA agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage. (Item 52) 52. The dsRNA agent of claim 51, wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 3' end of one strand. (Item 53) 53. The dsRNA agent of claim 52, wherein the strand is the antisense strand. (Item 54) 53. The dsRNA agent of claim 52, wherein the strand is the sense strand. (Item 55) 52. The dsRNA agent of claim 51, wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 5' end of one strand. (Item 56) 56. The dsRNA agent of claim 55, wherein the strand is the antisense strand. (Item 57) 56. The dsRNA agent of claim 55, wherein the strand is the sense strand. (Item 58) 52. The dsRNA agent of claim 51, wherein the phosphorothioate or methylphosphonate internucleotide linkages are at both the 5' and 3' ends of one strand. (Item 59) 31. The dsRNA agent according to Item 30, wherein the first base pair at the 5' end of the antisense strand of the duplex is an AU base pair. (Item 60) 31. The dsRNA agent according to item 30, wherein p'>0. (Item 61) 31. The dsRNA agent according to item 30, wherein p'=2. (Item 62) 62. The dsRNA agent of item 61, wherein q'=0, p=0, q=0, and the p' overhanging nucleotides are complementary to the target mRNA. (Item 63) 62. The dsRNA agent of item 61, wherein q'=0, p=0, q=0, and the p' overhanging nucleotides are non-complementary to the target mRNA. (Item 64) 31. The dsRNA agent of claim 30, wherein the sense strand has a total of 21 nucleotides and the antisense strand has a total of 23 nucleotides. (Item 65) At least one n p 31. The dsRNA agent according to item 30, wherein the ' is linked to adjacent nucleotides by a phosphorothioate linkage. (Item 66) All n p 66. The dsRNA agent according to item 65, wherein the ' is linked to adjacent nucleotides by a phosphorothioate linkage. (Item 67) 31. The dsRNA agent of claim 30, wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a modification. (Item 68) 68. The dsRNA agent of any one of paragraphs 30 to 67, wherein the ligand is conjugated to the 3' end of the sense strand of the dsRNA agent. (Item 69) 69. The dsRNA agent of claim 68, wherein the ligand is one or more N-acetylgalactosamine (GalNAc) derivatives attached through a monovalent, divalent, or trivalent branched linker. (Item 70) The ligand is [ka] 70. The dsRNA agent according to Item 69, wherein (Item 71) The ligand shown in the schematic diagram below [ka] It is conjugated with 71. The dsRNA agent of item 70, wherein X is O or S. (Item 72) 72. The dsRNA agent of item 71, wherein X is O. (Item 73) A double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, the dsRNA agent comprising a sense strand complementary to an antisense strand, the antisense strand comprising a region complementary to a portion of an mRNA encoding HSD17B13, and each strand being about 14 to about 30 nucleotides in length, the dsRNA agent having formula (III): Sense:5'n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j-N a -n q 3' Antisense: 3'n p '-N a '-(X'X'X') k -N b '-Y'Y'Y'-N b '-(Z'Z'Z') l -N a '-n q '5'(III) (In the formula, i, j, k, and l are each independently 0 or 1; p, p', q, and q' are each independently 0 to 6; each N a and N a ' represents an oligonucleotide sequence containing 0-25 nucleotides that are independently modified or unmodified, or a combination thereof, each sequence containing at least two differently modified nucleotides; each N b and N b ' represents an oligonucleotide sequence containing 0-10 nucleotides, independently, modified or unmodified, or a combination thereof; each n p , n p ',n q , and n q ' each may be present or absent and independently represent an overhanging nucleotide; XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent a motif having three identical modifications to three consecutive nucleotides, said modifications being 2'-O-methyl or 2'-fluoro modifications; N b The modification to Y is different from the modification to N b (The modifier for ' is different from the modifier for Y') is expressed as The dsRNA agent, wherein the sense strand is conjugated to at least one ligand. (Item 74) A double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, the dsRNA agent comprising a sense strand complementary to an antisense strand, the antisense strand comprising a region complementary to a portion of an mRNA encoding HSD17B13, and each strand being about 14 to about 30 nucleotides in length, the dsRNA agent having formula (III): Sense:5'n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3' Antisense: 3'n p '-N a '-(X'X'X') k -N b '-Y'Y'Y'-N b '-(Z'Z'Z') l -N a '-n q '5'(III) (In the formula, i, j, k, and l are each independently 0 or 1; each n p , n q , and n q ' each may be present or absent and independently represent an overhanging nucleotide; p, q, and q' are each independently 0 to 6; n p '>0 and at least one n p ' is linked to adjacent nucleotides by phosphorothioate linkages; each N a and N a ' represents an oligonucleotide sequence containing 0-25 nucleotides that are independently modified or unmodified, or a combination thereof, each sequence containing at least two differently modified nucleotides; each N b and N b' represents an oligonucleotide sequence containing 0-10 nucleotides, independently, modified or unmodified, or a combination thereof; XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent a motif having three identical modifications to three consecutive nucleotides, said modifications being 2'-O-methyl or 2'-fluoro modifications; N b The modification to N is different from the modification to Y. b (The modifier for ' is different from the modifier for Y') is represented by The dsRNA agent, wherein the sense strand is conjugated to at least one ligand. (Item 75) A double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, the dsRNA agent comprising a sense strand complementary to an antisense strand, the antisense strand comprising a region complementary to a portion of an mRNA encoding HSD17B13, and each strand being about 14 to about 30 nucleotides in length, the dsRNA agent having formula (III): Sense:5'n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3' Antisense: 3'n p '-N a '-(X'X'X') k -N b '-Y'Y'Y'-N b '-(Z'Z'Z') l -N a '-n q '5'(III) (In the formula, i, j, k, and l are each independently 0 or 1; each n p , n q , and nq ' each may be present or absent and independently represent an overhanging nucleotide; p, q, and q' are each independently 0 to 6; n p '>0 and at least one n p ' is linked to adjacent nucleotides by phosphorothioate linkages; each N a and N a ' represents an oligonucleotide sequence containing 0-25 nucleotides that are independently modified or unmodified, or a combination thereof, each sequence containing at least two differently modified nucleotides; each N b and N b ' represents an oligonucleotide sequence containing 0-10 nucleotides, independently, modified or unmodified, or a combination thereof; XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent a motif having three identical modifications to three consecutive nucleotides, said modifications being 2'-O-methyl or 2'-fluoro modifications; N b The modification to N is different from the modification to Y. b (The modifier for ' is different from the modifier for Y') is represented by the sense strand is conjugated to at least one ligand, the ligand being one or more GalNAc derivatives attached through a monovalent, divalent, or trivalent branched linker; dsRNA agents. (Item 76) A double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, the dsRNA agent comprising a sense strand complementary to an antisense strand, the antisense strand comprising a region complementary to a portion of an mRNA encoding HSD17B13, and each strand being about 14 to about 30 nucleotides in length, the dsRNA agent having formula (III): Sense:5'n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3' Antisense: 3'n p '-N a '-(X'X'X') k -N b '-Y'Y'Y'-N b '-(Z'Z'Z') l -N a '-n q '5'(III) (In the formula, i, j, k, and l are each independently 0 or 1; each n p , n q , and n q ' each may be present or absent and independently represent an overhanging nucleotide; p, q, and q' are each independently 0 to 6; n p '>0 and at least one n p ' is linked to adjacent nucleotides by phosphorothioate linkages; each N a and N a ' represents an oligonucleotide sequence containing 0-25 nucleotides that are independently modified or unmodified, or a combination thereof, each sequence containing at least two differently modified nucleotides; each N b and N b ' represents an oligonucleotide sequence containing 0-10 nucleotides, independently, modified or unmodified, or a combination thereof; XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent a motif having three identical modifications to three consecutive nucleotides, said modifications being 2'-O-methyl or 2'-fluoro modifications; N b The modification to N is different from the modification to Y. b (The modifier for ' is different from the modifier for Y') is represented by the sense strand comprises at least one phosphorothioate linkage; the sense strand is conjugated to at least one ligand, the ligand being one or more GalNAc derivatives attached through a monovalent, divalent, or trivalent branched linker; dsRNA agents. (Item 77) A double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, the dsRNA agent comprising a sense strand complementary to an antisense strand, the antisense strand comprising a region complementary to a portion of an mRNA encoding HSD17B13, and each strand being about 14 to about 30 nucleotides in length, the dsRNA agent having formula (III): Sense:5'n p -N a -YYY-N a -n q 3' Antisense: 3'n p '-N a '-Y'Y'Y'-N a '-n q '5'(IIIa) (In the formula, each n p , n q , and n q ' each may be present or absent and independently represent an overhanging nucleotide; p, q, and q' are each independently 0 to 6; n p '>0 and at least one n p ' is linked to adjacent nucleotides by phosphorothioate linkages; each N a and N a' represents an oligonucleotide sequence containing 0-25 nucleotides that are independently modified or unmodified, or a combination thereof, each sequence containing at least two differently modified nucleotides; YYY and Y'Y'Y' each independently represent a motif having three identical modifications to three consecutive nucleotides, said modifications being 2'-O-methyl or 2'-fluoro modifications. is represented by the sense strand comprises at least one phosphorothioate linkage; the sense strand is conjugated to at least one ligand, the ligand being one or more GalNAc derivatives attached through a monovalent, divalent, or trivalent branched linker; dsRNA agents. (Item 78) A double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, comprising a sense strand and an antisense strand forming a double-stranded region, the sense strand comprises at least 15 consecutive nucleotides that differ by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO: 1 or 2, and the antisense strand comprises at least 15 consecutive nucleotides that differ by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO: 8 or 9; substantially all of the nucleotides of the sense strand comprise a modification selected from the group consisting of a 2'-O-methyl modification and a 2'-fluoro modification; the sense strand comprises two phosphorothioate internucleotide linkages at the 5' end; substantially all of the nucleotides of the antisense strand comprise a modification selected from the group consisting of a 2'-O-methyl modification and a 2'-fluoro modification; the antisense strand comprises two phosphorothioate internucleotide linkages at the 5'-terminus and two phosphorothioate internucleotide linkages at the 3'-terminus; A dsRNA agent wherein the sense strand is conjugated at its 3' end to one or more GalNAc derivatives attached through a monovalent, divalent or trivalent branched linker. (Item 79) 79. The dsRNA agent of claim 78, wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand are modified nucleotides. (Item 80) 80. The dsRNA agent of any one of items 2, 30, and 73-79, wherein the region of complementarity comprises any one of the antisense sequences listed in any one of Tables 2, 3, 7, 8, 10, 11, or 13. (Item 81) 81. The dsRNA agent of any one of items 1 to 80, wherein the sense strand and the antisense strand comprise a nucleotide sequence selected from the group consisting of the nucleotide sequences of any one of the agents listed in any one of Tables 2, 3, 7, 8, 10, 11, or 13. (Item 82) 82. A cell containing the dsRNA agent of any one of items 1 to 81. (Item 83) 82. A vector encoding at least one strand of the dsRNA agent of any one of items 1 to 81. (Item 84) 82. A pharmaceutical composition for inhibiting the expression of the 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) gene, comprising the dsRNA agent according to any one of items 1 to 81. (Item 85) 85. The pharmaceutical composition of claim 84, wherein the agent is formulated in an unbuffered solution. (Item 86) 86. The pharmaceutical composition of claim 85, wherein the unbuffered solution is saline or water. (Item 87) 85. The pharmaceutical composition of claim 84, wherein the agent is formulated with a buffered solution. (Item 88) Item 89. The pharmaceutical composition of item 87, wherein the buffered solution comprises acetate, citrate, prolamine, carbonate, or phosphate, or any combination thereof. 88. The pharmaceutical composition of claim 87, wherein the buffered solution is phosphate buffered saline (PBS). (Item 90) 89. A method for inhibiting 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) expression in a cell, comprising contacting the cell with an agent according to any one of items 1 to 81 or a pharmaceutical composition according to any one of items 84 to 89, thereby inhibiting expression of HSD17B13 in the cell. (Item 91) Item 91. The method of item 90, wherein the cell is in a subject. (Item 92) 92. The method of claim 91, wherein the subject is a human. (Item 93) 93. The method of any one of items 90 to 92, wherein the HSD17B13 expression is inhibited by at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or is inhibited to below the level of detection of HSD17B13 expression. (Item 94) 94. The method of claim 93, wherein the human subject is suffering from a disease, disorder, or condition associated with HSD17B13. (Item 95) 95. The method of item 94, wherein the HSD17B13-associated disease, disorder, or condition is a chronic fibroinflammatory liver disease. (Item 96) Item 96. The method of item 95, wherein the chronic fibroinflammatory liver disease is associated with accumulation and / or enlargement of lipid droplets in the liver. (Item 97) 96. The method of item 95, wherein the chronic fibroinflammatory liver disease is selected from the group consisting of liver inflammation, hepatic fibrosis, nonalcoholic steatohepatitis (NASH), nonalcoholic fatty liver disease (NAFLD), cirrhosis, alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), HCV-associated cirrhosis, drug-induced liver injury, and hepatocellular necrosis. (Item 98) 87. A method for inhibiting expression of HSD17B13 in a subject, comprising administering to the subject a therapeutically effective amount of the dsRNA agent of any one of items 1 to 81 or the pharmaceutical composition of any one of items 84 to 89, thereby inhibiting expression of HSD17B13 in the subject. (Item 99) A method for treating a subject suffering from a disease, disorder, or condition associated with HSD17B13, comprising administering to the subject a therapeutically effective amount of an agent according to any one of items 1 to 81 or a pharmaceutical composition according to any one of items 84 to 89, thereby treating the subject suffering from a disease, disorder, or condition associated with HSD17B13. (Item 100) A method for preventing at least one symptom in a subject having a disease, disorder or condition for which reduced expression of the HSD17B13 gene is expected to be beneficial, comprising administering to the subject a prophylactically effective amount of an agent described in any one of items 1 to 31 or a pharmaceutical composition described in any one of items 34 to 39, thereby preventing at least one symptom in a subject having a disease, disorder or condition for which reduced expression of the HSD17B13 gene is expected to be beneficial. (Item 101) 87. A method for reducing the risk of developing chronic liver disease in a subject with steatosis, comprising administering to the subject a therapeutically effective amount of the dsRNA agent of any one of items 1 to 81 or the pharmaceutical composition of any one of items 84 to 89, thereby reducing the risk of developing chronic liver disease in the subject with steatosis. (Item 102) 87. A method for inhibiting the progression of steatosis to steatohepatitis in a subject suffering from steatosis, comprising administering to the subject a therapeutically effective amount of the dsRNA agent of any one of items 1 to 81 or the pharmaceutical composition of any one of items 84 to 89, thereby inhibiting the progression of steatosis to steatohepatitis in the subject. (Item 103) A method for inhibiting lipid droplet accumulation in the liver of a subject suffering from a disease, disorder, or condition associated with HSD17B13, comprising administering to the subject a therapeutically effective amount of the dsRNA agent of any one of items 1 to 81 or the pharmaceutical composition of any one of items 84 to 89, and a dsRNA agent targeting the PNPLA3 gene or a pharmaceutical composition comprising a dsRNA agent targeting the PNPLA3 gene, thereby inhibiting fat accumulation in the liver of the subject suffering from a disease, disorder, or condition associated with HSD17B13. (Item 104) 10. A method for treating a subject suffering from a disease, disorder, or condition associated with HSD17B13, comprising administering to the subject a therapeutically effective amount of the agent of any one of items 1 to 81 or the pharmaceutical composition of any one of items 84 to 89, and a dsRNA agent targeting the PNPLA3 gene or a pharmaceutical composition comprising a dsRNA agent targeting the PNPLA3 gene, thereby treating the subject suffering from the disease, disorder, or condition associated with HSD17B13. (Item 105) 106. A method for preventing at least one symptom in a subject having a disease, disorder, or condition for which reduced expression of the HSD17B13 gene is expected to be beneficial, the method comprising administering to the subject a prophylactically effective amount of the agent according to any one of Items 1 to 81 or the pharmaceutical composition according to any one of Items 84 to 89, and a dsRNA agent targeting the PNPLA3 gene or a pharmaceutical composition comprising a dsRNA agent targeting the PNPLA3 gene, thereby preventing at least one symptom in the subject having a disease, disorder, or condition for which reduced expression of the HSD17B13 gene is expected to be beneficial. 89. A method for reducing the risk of developing chronic liver disease in a subject with steatosis, comprising administering to the subject a therapeutically effective amount of the dsRNA agent of any one of items 1 to 81 or the pharmaceutical composition of any one of items 84 to 89, and a dsRNA agent targeting the PNPLA3 gene or a pharmaceutical composition comprising a dsRNA agent targeting the PNPLA3 gene, thereby reducing the risk of developing chronic liver disease in the subject with steatosis. (Item 107) 89. A method for inhibiting the progression of steatosis to steatohepatitis in a subject suffering from steatosis, comprising administering to the subject a therapeutically effective amount of the dsRNA agent of any one of items 1 to 81 or the pharmaceutical composition of any one of items 84 to 89, and a dsRNA agent targeting the PNPLA3 gene or a pharmaceutical composition comprising a dsRNA agent targeting the PNPLA3 gene, thereby inhibiting the progression of steatosis to steatohepatitis in the subject. (Item 108) 108. The method of any one of items 91 to 107, wherein administering the dsRNA agent or the pharmaceutical composition to the subject causes a decrease in HSD17B13 enzyme activity, a decrease in HSD17B13 protein accumulation, a decrease in PNPLA3 enzyme activity, a decrease in PNPLA3 protein accumulation, and / or a decrease in fat accumulation and / or an increase in lipid droplets in the liver of the subject. (Item 109) 109. The method of any one of items 99 to 108, wherein the HSD17B13-associated disease, disorder, or condition is a chronic fibroinflammatory liver disease. (Item 110) Item 109. The method of item 109, wherein the chronic fibroinflammatory liver disease is associated with accumulation and / or enlargement of lipid droplets in the liver. (Item 111) 109. The method of claim 109, wherein the chronic fibroinflammatory liver disease is selected from the group consisting of fat accumulation in the liver, liver inflammation, hepatic fibrosis, fatty liver disease (steatosis), non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cirrhosis, alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), HCV-associated cirrhosis, drug-induced liver injury, and hepatocellular necrosis. (Item 112) Item 112. The method of item 111, wherein the chronic fibroinflammatory liver disease is nonalcoholic steatohepatitis (NASH). (Item 113) 113. The method of any one of items 91 to 112, wherein the subject is obese. (Item 114) 114. The method of any one of items 91 to 113, further comprising administering an additional therapeutic agent to the subject. (Item 115) 115. The method of any one of items 91 to 114, wherein the dsRNA agent is administered to the subject 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. (Item 116) 116. The method of any one of items 91 to 115, wherein the agent is administered to the subject intravenously, intramuscularly, or subcutaneously. (Item 117) 117. The method of any one of items 91 to 116, further comprising determining the level of HSD17B13 in the subject. (Item 118) 1. A double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, comprising: comprising a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises the nucleotide sequence of any one of the agents in any one of Tables 2, 3, 7, 8, 10, 11, or 13, and the antisense strand comprises the nucleotide sequence of any one of the agents in any one of Tables 2, 3, 7, 8, 10, 11, or 13; substantially all of the nucleotides of the sense strand and substantially all of the nucleotides of the antisense strand are modified nucleotides; The dsRNA agent, wherein the dsRNA agent is conjugated to a ligand. [Brief explanation of the drawings]

[0097] [Figure 1] Figure 1 shows that HSD17B13 rs72613567:TA is associated with reduced risk of alcoholic and non-alcoholic liver disease phenotypes. Specifically, Figure 1 shows that HSD17B13 rs72613567:TA was associated with allele dosage-dependent lower odds of various chronic liver diseases. Specifically, allele dosage-dependent effects were observed for both alcoholic and non-alcoholic liver disease, cirrhosis, and hepatocellular carcinoma. Odds ratios were calculated using logistic regression, adjusting for age, sex, BMI, and self-reported ethnicity.

[0098] [Figure 2] Figure 2A is a graph showing the effect of a single dose of AD-288917 in mice expressing human HSD17B13, and Figure 2B is a graph showing the effect of a single dose of AD-288917 in cynomolgus monkeys. DETAILED DESCRIPTION OF THE INVENTION

[0099] The present invention provides an iRNA composition that causes RNA-induced silencing complex (RISC)-mediated cleavage of the RNA transcript of HSD17B13 gene.The HSD17B13 gene can be present in a cell, for example, in a cell of a subject, such as a human.The present invention also provides a method for using the iRNA composition of the present invention to inhibit the expression of HSD17B13 gene, and to treat subjects who are expected to benefit from inhibiting or reducing the expression of HSD17B13 gene, for example, subjects who are expected to benefit from reducing liver inflammation, for example, subjects who suffer from or are prone to HSD17B13-related diseases, disorders, or conditions, such as liver fibrosis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), liver cirrhosis, HCV-related cirrhosis, drug-induced liver injury, and liver cell necrosis.

[0100] The iRNA of the present invention targeting HSD17B13 may be about 30 nucleotides in length or less, for example, 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-26, 19-27, 19-28, 19-29, 19-30, 19-29, 19-28, 19-27, 19-26, 19-26, 19-27, 19-28 ...9, 19-30, 19-29, 19-28, 19-27, 19-26, 19-26, 19-26, 19-27, 19-28, 19-29, 19-30, 19-29, 19-28, 1 The RNA strand may comprise an RNA strand (antisense strand) having a region that is 1 to 25, 19 to 24, 19 to 23, 19 to 22, 19 to 21, 19 to 20, 20 to 30, 20 to 29, 20 to 28, 20 to 27, 20 to 26, 20 to 25, 20 to 24, 20 to 23, 20 to 22, 20 to 21, 21 to 30, 21 to 29, 21 to 28, 21 to 27, 21 to 26, 21 to 25, 21 to 24, 21 to 23, or 21 to 22 nucleotides in length, which region is substantially complementary to at least a portion of an mRNA transcript of the HSD17B13 gene.

[0101] In some embodiments, one or both strands of a double-stranded RNAi agent of the invention are up to 66 nucleotides in length, e.g., 36-66, 26-36, 25-36, 31-60, 22-43, or 27-53 nucleotides in length, and have a region of at least 19 contiguous nucleotides that are substantially complementary to at least a portion of an mRNA transcript of the HSD17B13 gene. In some embodiments, such iRNA agents with longer antisense strands may include a second RNA strand (sense strand) 20-60 nucleotides in length, where the sense and antisense strands form a duplex of 18-30 contiguous nucleotides.

[0102] The use of the iRNA agents described herein allows for the targeted degradation of the mRNA of the HSD17B13 gene in mammals.

[0103] In particular, very low doses of iRNA can specifically and effectively mediate RNA interference (RNAi), resulting in the significant inhibition of the expression of HSD17B13 gene.Therefore, the method and composition that comprise these iRNA are useful for treating the subjects that are expected to be beneficial in inhibiting or reducing the expression of HSD17B13 gene, for example, the subjects that are expected to be beneficial in reducing liver inflammation, for example, the subjects that suffer from or are prone to suffer from HSD17B13-related diseases, disorders or conditions, for example, the subjects that suffer from or are prone to suffer from liver fibrosis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), liver cirrhosis, HCV-related cirrhosis, drug-induced liver injury and liver cell necrosis.

[0104] The detailed description below discloses how to make and use compositions containing iRNA to inhibit expression of the HSD17B13 gene, as well as compositions and methods for treating subjects with diseases and disorders in which inhibition and / or reduction of expression of this gene would be beneficial. I. Definition

[0105] In order that the present invention may be more readily understood, certain terms are first defined. Furthermore, it should be noted that whenever a value or range of values ​​for a parameter is listed, it is intended that values ​​and ranges between the listed values ​​are also part of the present invention.

[0106] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element, e.g., a plurality of elements.

[0107] The term "including" is used herein to mean, and is used interchangeably with, the phrase "including but not limited to."

[0108] The term "or" is used herein to mean, and is used interchangeably with, the term "and / or," unless context clearly indicates otherwise.

[0109] The term "about" is used herein to mean within a range of typical acceptance in the art. For example, "about" can be understood as about two standard deviations from the mean. In certain embodiments, about means ±10%. In certain embodiments, about means ±5%. When about is before a series or range, it is understood that "about" can modify each of the numbers in the series or range.

[0110] The term "HSD17B13," also known as "hydroxysteroid 17-beta dehydrogenase 13," "short-chain dehydrogenase / reductase family 16C member," "short-chain dehydrogenase / reductase 9," "17-beta-HSD13," "17β-HSD13," "SDR16C3," "SCDR9," "short-chain dehydrogenase / reductase family 16C, member 3," "hydroxysteroid (17-beta) dehydrogenase 13," "17-beta-hydroxysteroid dehydrogenase 13," "17-beta hydroxysteroid dehydrogenase," "HMFN0376," and "NIIL497," refers to the well-known gene encoding 17β-hydroxysteroid dehydrogenase type 13 protein from any vertebrate or mammalian source, including, but not limited to, human, bovine, chicken, rodent, mouse, rat, pig, sheep, primate, monkey, and guinea pig, unless otherwise specified.

[0111] The term also refers to fragments and variants of native HSD17B13 that maintain at least one in vivo or in vitro activity of native HSD17B13. The term encompasses full-length, unprocessed precursor forms of HSD17B13 as well as mature forms and forms resulting from proteolytic processing resulting from post-translational cleavage of the signal peptide.

[0112] Two variants of the human HSD17B13 gene, variant A (or transcript A) and variant B (or transcript B), have previously been identified. Transcript A contains all seven exons of the HSD17B13 gene, while transcript B skips exon 2. The nucleotide and amino acid sequences of human HSD17B13 variant A can be found, for example, in GenBank Reference Sequence: NM_178135.4; SEQ ID NO: 1), and the nucleotide and amino acid sequences of human HSD17B13 variant B can be found, for example, in GenBank Reference Sequence: NM_001136230.2; SEQ ID NO: 2. As described in U.S. Patent Application No. 15 / 875,514, filed January 19, 2018, and PCT Application No. PCT / US2018 / 014357, filed January 19, 2018, the entire contents of each of which are incorporated herein by reference, six additional expressed HSD17B13 transcripts (C-H, SEQ ID NOS: 17, 18, 19, 20, 21, and 22, respectively) have been identified. Transcript C skips exon 6 compared to transcript A. Transcript D inserts a guanine 3' of exon 6 compared to transcript A, resulting in a frameshift in exon 7 and premature truncation of exon 7. Transcript E contains an additional exon between exons 3 and 4 compared to transcript A. Transcript F, which is expressed only in HSD17B13 rs72613567 variant carriers, has a readthrough from exon 6 to intron 6 compared to transcript A. Transcript G has a skip of exon 2 and an insertion of guanine 3' of exon 6 compared to transcript A, resulting in a frameshift in exon 7 and a premature truncation of exon 7. Transcript H has an additional exon between exons 3 and 4 and an insertion of guanine 3' of exon 6 compared to transcript A, resulting in a frameshift in exon 7 and a premature truncation of exon 7.

[0113] One additional HSD17B13 transcript (F', SEQ ID NO: 23) that is expressed at low levels has also been identified. Like transcript F, transcript F' also contains a readthrough from exon 6 to intron 6 compared to transcript A, but in contrast to transcript F, the readthrough does not contain the thymine insertion present in the HSD17B13 rs72613567 variant gene. The nucleotide positions of the exons within the HSD17B13 gene for each transcript are provided below.

[0114] SEQ ID NO: 15 is the nucleotide sequence of the HSD17B13 wild-type genomic sequence (human genome assembly GRCh38), and SEQ ID NO: 16 is the nucleotide sequence of the HSD17B13 genomic sequence variant (human genome assembly GRCh38; rs72613567-T insertion in chr4: 87310241-87310240): T insertion at position 12666. Nucleotide positions in SEQ ID NO: 15 for exons of the HSD17B13 transcript that are more prevalent in subjects homozygous for the wild-type HSD17B13 gene. [Table 15] rs72613567 Nucleotide position in SEQ ID NO: 16 for the exon of the HSD17B13 transcript that is more prevalent in subjects homozygous for the HSD17B13 variant gene (insertion of T at position 12666). [Table 16]

[0115] Two variants of the mouse HSD17B13 gene exist; the nucleotide and amino acid sequences of mouse Hsd17b13, transcript variant 1 can be found, for example, in GenBank Reference Sequence: NM_001163486.1 (SEQ ID NO: 3), and the nucleotide and amino acid sequences of mouse Hsd17b13, transcript variant 2 can be found, for example, in GenBank Reference Sequence: NM_198030.2 (SEQ ID NO: 4). The nucleotide and amino acid sequences of the rat Hsd17b13 gene can be found, for example, in GenBank Reference Sequence: NM_001009684.1 (SEQ ID NO: 5). The nucleotide and amino acid sequences of the Macaca mulatta HSD17B13 gene can be found, for example, in GenBank Reference Sequence: XM_015138766.1 (SEQ ID NO: 6). The nucleotide and amino acid sequence of the Macaca fascicularis HSD17B13 gene can be found, for example, in GenBank Reference Sequence: XM_005555367.2; SEQ ID NO: 7).

[0116] Further examples of HSD17B13 mRNA sequences are readily available using publicly available databases such as GenBank, UniProt, and OMIM.

[0117] The term "HSD17B13" as used herein also refers to a specific polypeptide expressed in cells due to naturally occurring DNA sequence variations of the HSD17B13 gene, such as single nucleotide polymorphisms in the HSD17B13 gene.A number of SNPs within the HSD17B13 gene have been identified, and can be found, for example, in NCBI dbSNP (see, for example, www.ncbi.nlm.nih.gov / snp).

[0118] As used herein, "target sequence" refers to a contiguous portion of the nucleotide sequence of an mRNA molecule formed during transcription of the HSD17B13 gene, including mRNA that is the product of RNA processing of a primary transcript. In one embodiment, the target portion of the sequence is at least sufficiently long to serve as a substrate for iRNA-directed cleavage at or near that portion of the nucleotide sequence of an mRNA molecule formed during transcription of the HSD17B13 gene.

[0119] The target sequence of the HSD17B13 gene can be about 9 to 36 nucleotides in length, for example, about 15 to 30 nucleotides in length. For example, the target sequence can be about 15 to 30 nucleotides, 15 to 29, 15 to 28, 15 to 27, 15 to 26, 15 to 25, 15 to 24, 15 to 23, 15 to 22, 15 to 21, 15 to 20, 15 to 19, 15 to 18, 15 to 17, 18 to 30, 18 to 29, 18 to 28, 18 to 27, 18 to 26, 18 to 25, 18 to 24, 18 to 23, 18 to 22, 18 to 21, 18 to 20, 19 to 30, 19 to 29, 19 It can be up to 28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides in length. Ranges and lengths between the above ranges and lengths are also contemplated as part of the invention.

[0120] As used herein, the term "strand made up of a sequence" refers to an oligonucleotide made up of a strand of nucleotides described by a sequence referenced using standard nucleotide nomenclature.

[0121] " G ", " C ", " A ", " T " and " U " generally represent the nucleotide that contains guanine, cytosine, adenine, thymidine and uracil as base, respectively. However, it should be understood that the term " ribonucleotide " or " nucleotide " can also refer to the modified nucleotide or substitute replacement portion, which will be described in more detail below (see, for example, Table 1). Those skilled in the art are well aware that guanine, cytosine, adenine and uracil can be replaced with other portions without substantially changing the base pairing properties of the oligonucleotide that contains the nucleotide with such replacement portion. For example, and without being limited thereto, the nucleotide that contains inosine as base can base pair with the nucleotide that contains adenine, cytosine or uracil. Therefore, the nucleotide that contains uracil, guanine or adenine in the nucleotide sequence of the dsRNA characterized in the present invention can be replaced with, for example, the nucleotide that contains inosine. In another example, adenine and cytosine anywhere within an oligonucleotide can be replaced with guanine and uracil, respectively, to form a GU wobble base pair with the target mRNA. Sequences containing such replacements are suitable for the compositions and methods featured in this invention.

[0122] The terms " iRNA ", " RNAi agent ", " iRNA agent ", " RNA interference agent " are used interchangeably herein and refer to an agent that contains RNA, a term defined herein, and mediates the targeted cleavage of RNA transcripts via the RNA-induced silencing complex (RISC) pathway. iRNA directs the sequence-specific degradation of mRNA through a process known as RNA interference (RNAi). iRNA modulates, for example, inhibits, the expression of HSD17B13 gene in cells, for example, cells in a subject, such as a mammalian subject.

[0123] In one embodiment, the RNAi agent of the present invention comprises a single-stranded RNA that interacts with a target RNA sequence, e.g., an HSD17B13 target mRNA sequence, to direct cleavage of the target RNA. Without wishing to be bound by theory, it is believed that long double-stranded RNA introduced into cells is degraded into siRNAs by a type III endonuclease known as Dicer (Sharp et al. (2001) Genes Dev. 15: 485). Dicer is a ribonuclease that degrades ribonuclease (RNase I). The dsRNA is a cleavage-III-like enzyme that processes dsRNA into 19-23 base pair small interfering RNAs with characteristic 2-base 3' overhangs (Bernstein, et al., (2001) Nature 409: 363). The siRNAs are then ligated into the RNA-induced silencing complex (RNA-induced silencing complex). The siRNA is incorporated into the ISC (Intersense Sequence Binding Site), where one or more helicases unwind the siRNA duplex, allowing the complementary antisense strand to guide target recognition (Nykanen, et al., (2001) Cell 107: 309). When this happens, one or more endonucleases in RISC will cleave the target, inducing silencing (Elbashir, et al., (2001) Genes Dev. 15: 188).Therefore, in one aspect, the present invention relates to the single-stranded RNA (sssiRNA) that is produced in cells and promotes the formation of RISC complex, resulting in the silencing of target gene, i.e., HSD17B13 gene.Therefore, the term " siRNA " is also used herein to refer to the above-mentioned RNAi.

[0124] In another embodiment, the RNAi agent can be a single-stranded RNAi agent introduced into a cell or organism to inhibit target mRNA. Single-stranded RNAi agents (ssRNAi) bind to the RISC endonuclease, Argonaute 2, which then cleaves the target mRNA. Single-stranded siRNAs are generally 15-30 nucleotides and chemically modified. The design and testing of single-stranded RNAi agents is described in U.S. Patent No. 8,101,348 and Lima et al., (2012) Cell 150: 883-894, the entire contents of each of which are hereby incorporated by reference. The RNAi agents described herein can be used in a variety of applications. Any of the antisense nucleotide sequences can be used as single-stranded siRNAs described herein or chemically modified by the methods described in Lima et al., (2012) Cell 150;:883-894.

[0125] In another embodiment, the "iRNA" for use in the compositions and methods of the present invention is double-stranded RNA, and is referred to herein as a "double-stranded RNAi agent," a "double-stranded RNA (dsRNA) molecule," a "dsRNA agent," or a "dsRNA." The term "dsRNA" refers to a complex of ribonucleic acid molecules having a duplex structure composed of two antiparallel and substantially complementary nucleic acid strands, which are referred to as having "sense" and "antisense" orientations relative to the target RNA, i.e., the HSD17B13 gene. In some embodiments of the present invention, double-stranded RNA (dsRNA) induces the degradation of target RNA, e.g., mRNA, through a post-transcriptional gene silencing mechanism, referred to herein as RNA interference or RNAi.

[0126] Generally, the majority of the nucleotides in each strand of dsRNA molecule are ribonucleotides; however, as described in detail herein, each strand or both strands can also contain one or more non-ribonucleotides, such as deoxyribonucleotides and / or modified nucleotides.Furthermore, as used herein, "RNAi agent" can include ribonucleotides with chemical modifications; RNAi agent can contain substantial modifications in multiple nucleotides.As used herein, the term "modified nucleotide" refers to the nucleotide that has independently modified sugar moiety, modified internucleotide linkage, and / or modified nucleobase.Therefore, the term modified nucleotide includes, for example, the substitution, addition, or removal of functional groups or atoms to internucleoside linkage, sugar moiety, or nucleobase.The modifications suitable for use in the agent of the present invention include all types of modifications disclosed herein or known in the art.All such modifications used in siRNA type molecules are included in "RNAi agent" for the purpose of this specification and claims.

[0127] The duplex region can be any length that allows for specific degradation of the desired target RNA through the RISC pathway, and can be from about 9 to 36 base pairs in length, e.g., about 15-30 base pairs in length, e.g., about 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 base pairs in length, e.g., about 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-19, 15-20, 15-21, 15-22, 15-23, 15-24, 15-25, 15-26, 15-27, 15-28, 15-29, 15-30, 15-31, 15-32, 15-33, 15-34, 15-35, 15-36, 15-37, 15-38, 15-39, 15-40, 15-41, 15-42, 15-43, 15-44, 15-45, 15-46, 15-47, 15-48, 15-49, 15-50, 15-51, 15 7, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-22 and the like, may range in length from 0, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs in length, etc. Ranges and lengths between the above ranges and lengths are also contemplated as being part of the invention.

[0128] The two strands forming a duplex structure may be different parts of a larger RNA molecule or may be separate RNA molecules. When the two strands are part of a larger molecule and are therefore connected by an uninterrupted chain of nucleotides between the 3' end of one strand and the 5' end of the other strand forming the duplex structure, the connected RNA strands are called "hairpin loops." A hairpin loop may contain at least one unpaired nucleotide. In some embodiments, a hairpin loop may contain at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 20, at least 23 or more unpaired nucleotides.

[0129] When the two substantially complementary strands of dsRNA are composed of separate RNA molecules, these molecules do not need to be connected by a covalent bond, but can be.When the two strands are covalently connected by means other than an uninterrupted chain of nucleotides between the 3'-end of one strand and the 5'-end of each other strand that form a double-stranded structure, the connecting structure is called a "linker".RNA strands can have the same or different number of nucleotides.The maximum number of base pairs is the number of nucleotides of the shortest strand of dsRNA minus any overhang that exists in the double strand.In addition to the double-stranded structure, RNAi can include one or more nucleotide overhangs.

[0130] In one embodiment, the RNAi agent of the present invention is a dsRNA, each strand of which comprises fewer than 30 nucleotides, e.g., 17-27, 19-27, 17-25, 19-25, or 19-23, that interact with a target RNA sequence, e.g., an HSD17B13 target mRNA sequence, to direct cleavage of the target RNA. In another embodiment, the RNAi agent of the present invention is a dsRNA, each strand of which comprises 19-23 nucleotides that interact with a target RNA sequence, e.g., an HSD17B13 target mRNA sequence, to direct cleavage of the target RNA. In one embodiment, the sense strand is 21 nucleotides in length. In another embodiment, the antisense strand is 23 nucleotides in length.

[0131] As used herein, the term "nucleotide overhang" refers to at least one unpaired nucleotide that protrudes from the double-stranded structure of an iRNA, such as a dsRNA. For example, a nucleotide overhang exists when the 3'-end of one strand of a dsRNA extends beyond the 5'-end of the other strand, or vice versa. A dsRNA can contain an overhang of at least one nucleotide; alternatively, the overhang can contain at least two nucleotides, at least three nucleotides, at least four nucleotides, at least five nucleotides, or more nucleotides. The nucleotide overhang can comprise or consist of nucleotide / nucleoside analogs, including deoxynucleotides / nucleosides. The overhang can be in the sense strand, the antisense strand, or any combination thereof. Furthermore, the nucleotide of the overhang can be present at the 5'-end, 3'-end, or both ends of either the antisense or sense strand of a dsRNA.

[0132] In one embodiment, the antisense strand of the dsRNA has an overhang of 1 to 10 nucleotides, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides, at the 3'-end and / or 5'-end. In one embodiment, the sense strand of the dsRNA has an overhang of 1 to 10 nucleotides, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides, at the 3'-end and / or 5'-end. In another embodiment, one or more of the nucleotides in the overhang are replaced with a nucleoside thiophosphate.

[0133] In certain embodiments, the overhang of the sense strand or the antisense strand, or both, can comprise an extended length greater than 10 nucleotides, e.g., 10-30 nucleotides, 10-25 nucleotides, 10-20 nucleotides, or 10-15 nucleotides in length. In certain embodiments, the extended overhang is on the sense strand of the duplex. In certain embodiments, the extended overhang is at the 3'-end of the sense strand of the duplex. In certain embodiments, the extended overhang is at the 5'-end of the sense strand of the duplex. In certain embodiments, the extended overhang is on the antisense strand of the duplex. In certain embodiments, the extended overhang is at the 3'-end of the antisense strand of the duplex. In certain embodiments, the extended overhang is at the 5'-end of the antisense strand of the duplex. In certain embodiments, one or more of the nucleotides of the extended overhang are replaced with a nucleoside thiophosphate.

[0134] The term "blunt" or "blunt-ended" as used herein with respect to dsRNA means that there are no unpaired nucleotides or nucleotide analogs at a given end of the dsRNA, i.e., there are no nucleotide overhangs. One or both ends of the dsRNA may be blunt-ended. If both ends of the dsRNA are blunt-ended, the dsRNA is said to be blunt-ended. For clarity, a "blunt-ended" dsRNA is a dsRNA that has both ends blunt-ended, i.e., there are no nucleotide overhangs at either end of the molecule. Such molecules are often double-stranded throughout their entire length.

[0135] The term "antisense strand" or "guide strand" refers to the strand of an iRNA, e.g., a dsRNA, that includes a region that is substantially complementary to a target sequence, e.g., HSD17B13 mRNA.

[0136] As used herein, the term "complementary region" refers to a region on the antisense strand that is substantially complementary to a sequence, for example, a target sequence, for example, the HSD17B13 nucleotide sequence defined herein.If the complementary region is not completely complementary to the target sequence, mismatches can be located in the internal or terminal regions of the molecule.Generally, the most tolerable mismatches are located in the terminal regions, for example, within 5, 4, 3, or 2 nucleotides from the 5' and / or 3' end of the iRNA.

[0137] The terms "sense strand" or "passenger strand," as used herein, refer to the strand of an iRNA that includes a region that is substantially complementary to a region of the antisense strand, as those terms are defined herein.

[0138] As used herein, the term "cleavage region" refers to the region located immediately adjacent to the cleavage site. The cleavage site is the site where cleavage occurs on the target. In some embodiments, the cleavage region comprises 3 bases immediately adjacent to the cleavage site on either end of the cleavage site. In some embodiments, the cleavage region comprises 2 bases immediately adjacent to the cleavage site on either end of the cleavage site. In some embodiments, the cleavage site is specifically located at the site where nucleotides 10 and 11 of the antisense strand bind, and the cleavage region comprises nucleotides 11, 12 and 13.

[0139] As used herein, unless otherwise specified, the term "complementary," when used to describe a first nucleotide sequence in the context of a second nucleotide sequence, refers to the ability of an oligonucleotide or polynucleotide comprising the first nucleotide sequence to hybridize to an oligonucleotide or polynucleotide comprising the second nucleotide sequence under certain conditions to form a duplex structure, as would be understood by one of skill in the art. Such conditions may be, for example, stringent conditions, which may include 400 mM NaCl, 40 mM PIPES, pH 6.4, 1 mM EDTA, at 50°C or 70°C for 12-16 hours, followed by washing (see, e.g., "Molecular Cloning: A Laboratory Manual," Sambrook, et al. (1989) Cold Spring Harbor Laboratory Press). The term "complementary" refers to a hybridization of a first nucleotide sequence with a second nucleotide sequence under certain conditions to form a duplex structure, as would be understood by one of skill in the art. Such conditions may be, for example, stringent conditions, which may include 400 mM NaCl, 40 mM PIPES, pH 6.4, 1 mM EDTA, at 50°C or 70°C for 12-16 hours, followed by washing (see, e.g., "Molecular Cloning: A Laboratory Manual," Sambrook, et al. (1989) Cold Spring Harbor Laboratory Press). Other conditions, such as conditions that are logically relevant, can be applied. Those skilled in the art can determine the set of conditions that are most suitable for testing the complementarity of two sequences depending on the final application of the hybridized nucleotides.

[0140] A complementary sequence within an iRNA, such as a dsRNA described herein, includes base pairing across the entire length of one or both of the nucleotide sequences of an oligonucleotide or polynucleotide comprising a first nucleotide sequence and an oligonucleotide or polynucleotide comprising a second nucleotide sequence. Such sequences may be referred to herein as "fully complementary" to each other. However, when a first sequence is referred to herein as "substantially complementary" to a second sequence, the two sequences may be fully complementary, or may form one or more, but generally no more than 5, 4, 3, or 2, mismatched base pairs when the duplex is hybridized up to 30 base pairs, while retaining the ability to hybridize under conditions most relevant to their ultimate application, such as inhibiting gene expression via the RISC pathway. However, if two oligonucleotides are designed to form one or more single-stranded overhangs upon hybridization, such overhangs should not be considered mismatches in determining complementarity. For example, a dsRNA composed of one oligonucleotide 21 nucleotides in length and another oligonucleotide 23 nucleotides in length, where the longer oligonucleotide contains a 21 nucleotide sequence that is perfectly complementary to the shorter oligonucleotide, may still be referred to as "fully complementary" for purposes described herein.

[0141] "Complementary" sequences, as used herein, may contain or be formed entirely from non-Watson-Crick base pairs and / or base pairs formed from non-naturally occurring modified nucleotides, so long as the above requirements for hybridization are met. Such non-Watson-Crick base pairs include, but are not limited to, G:U wobble or Hoogsteen base pairing.

[0142] The terms "complementary," "fully complementary," and "substantially complementary" may be used herein with reference to matching bases between the sense and antisense strands of a dsRNA or between the antisense strand of an iRNA agent and a target sequence, as understood from the context in which they are used.

[0143] As used herein, a polynucleotide that is "substantially complementary to at least a portion" of a messenger RNA (mRNA) refers to a polynucleotide that is substantially complementary to a contiguous portion of an mRNA of interest (e.g., an mRNA encoding HSD17B13). For example, a polynucleotide is complementary to at least a portion of an HSD17B13 mRNA if the sequence is substantially complementary to an uninterrupted portion of the mRNA encoding HSD17B13.

[0144] Thus, in some embodiments, the antisense strand polynucleotides disclosed herein are fully complementary to the target HSD17B13 sequence. In other embodiments, the antisense strand polynucleotides disclosed herein are substantially complementary to the target HSD17B13 sequence, and comprise a contiguous nucleotide sequence over its entire length that is at least about 80% complementary, for example, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% complementary to the equivalent region of the nucleotide sequence of SEQ ID NO: 1, or a fragment of SEQ ID NO: 1.

[0145] In one embodiment, an RNAi agent of the invention comprises a sense strand substantially complementary to an antisense polynucleotide, wherein the antisense polynucleotide is complementary to a target HSD17B13 sequence, and the sense strand polynucleotide comprises a contiguous nucleotide sequence over its entire length that is at least about 80% complementary, e.g., about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% complementary to an equivalent region of the nucleotide sequence of SEQ ID NO:8, or a fragment of any one of SEQ ID NO:8.

[0146] In some embodiments, the iRNA of the invention comprises an antisense strand that is substantially complementary to a target HSD17B13 sequence and comprises a contiguous nucleotide sequence over its entire length that is at least about 80% complementary, e.g., about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% complementary, or 100% complementary, to the equivalent region of the nucleotide sequence of any one of the sense strands in any one of Tables 2, 3, 7, 8, 10, 11, or 13, or a fragment of any one of the sense strands in any one of Tables 2, 3, 7, 8, 10, 11, or 13.

[0147] The term "inhibiting," as used herein, is used interchangeably with "reducing," "silencing," "downregulating," "suppressing," and other similar terms, and includes any level of inhibition.

[0148] The phrase "inhibiting expression of the HSD17B13 gene," as used herein, includes inhibiting the expression of any HSD17B13 gene that encodes an HSD17B13 protein (e.g., a mouse HSD17B13 gene, a rat HSD17B13 gene, a monkey HSD17B13 gene, or a human HSD17B13 gene, etc.), as well as variants or mutants of the HSD17B13 gene.

[0149] "Inhibiting expression of the HSD17B13 gene" includes any level of inhibition of the HSD17B13 gene, for example, at least partial suppression of expression of the HSD17B13 gene, for example, at least about 20% inhibition, etc. In certain embodiments, the inhibition is at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% inhibition.

[0150] The expression of HSD17B13 gene can be evaluated based on the level of any variable related to HSD17B13 gene expression, for example, HSD17B13 mRNA level or HSD17B13 protein level.The expression of HSD17B13 gene can also be evaluated indirectly based on, for example, the level of circulating alanine aminotransferase (ALT) or the enzyme activity of HSD17B13 in tissue samples such as liver samples.Inhibition can be evaluated by the absolute level of one or more of these variables, or the relative level compared with the control level.The control level can be any type of control level used in the art, for example, the baseline level before administration, or the level determined from the same subject, cell, or sample that is not treated or treated with a control (for example, a buffer-only control or an inactive agent control, etc.).

[0151] In one embodiment, at least partial suppression of expression of the HSD17B13 gene is assessed by a reduction in the amount of HSD17B13 mRNA that can be isolated from or detected in a first cell or group of cells in which the HSD17B13 gene is transcribed and that has been treated to inhibit expression of the HSD17B13 gene, compared to a second cell or group of cells (control cells) that is substantially identical to the first cell or group of cells but has not been so treated.

[0152] The degree of inhibition is

number

[0153] The phrase "contacting a cell with an RNAi agent" such as dsRNA as used herein includes contacting a cell by any possible means. Contacting a cell with an RNAi agent includes contacting a cell with an iRNA in vitro or contacting a cell with an iRNA in vivo. Contacting can be performed directly or indirectly. Thus, for example, the RNAi agent can be physically contacted with a cell by performing a method individually, or the RNAi agent can be placed in a situation that allows or causes the RNAi agent to subsequently contact a cell.

[0154] In vitro contacting of cells can be carried out, for example, by incubating cells with RNAi agent.In vivo contacting of cells can be carried out, for example, by injecting RNAi agent into the tissue where cells are located or nearby, or by injecting RNAi agent into another area, for example, bloodstream or subcutaneous space, so that the agent can then reach the tissue where the cells to be contacted are located.For example, RNAi agent can contain and / or be coupled with a ligand, such as GalNAc3, that directs RNAi agent to target site, for example, liver.Combination of in vitro and in vivo contacting methods is also possible.For example, cell can be contacted with RNAi agent in vitro, and then transplanted into subject.

[0155] In one embodiment, contacting a cell with an iRNA includes "introducing" or "delivering the iRNA to a cell" by facilitating or causing uptake or absorption into the cell. Absorption or uptake of the iRNA can occur through unassisted diffusive or active cellular processes, or by auxiliary agents or devices. Introduction of the iRNA into a cell can be in vitro and / or in vivo. For example, for in vivo introduction, the iRNA can be injected at a tissue site or administered systemically. In vivo delivery can also be achieved via beta-glucan delivery systems, such as those described in U.S. Pat. Nos. 5,032,401 and 5,607,677, and U.S. Publication No. 2005 / 0281781, the entire contents of which are hereby incorporated by reference. In vitro introduction into a cell includes methods known in the art, such as electroporation and lipofection. Other techniques are described herein below and / or known in the art.

[0156] The term "lipid nanoparticle" or "LNP" refers to a vesicle composed of a lipid layer encapsulating a pharmaceutically active molecule, such as a nucleic acid molecule, e.g., an iRNA or a plasmid into which the iRNA is transcribed. LNPs are described, for example, in U.S. Patent Nos. 6,858,225, 6,815,432, 8,158,601, and 8,058,069, the entire contents of which are hereby incorporated by reference.

[0157] As used herein, a "subject" is an animal such as a mammal, including a primate (e.g., a human, a non-human primate such as a monkey, and a chimpanzee), a non-primate (e.g., a cow, a pig, a camel, a llama, a horse, a goat, a rabbit, a sheep, a hamster, a guinea pig, a cat, a dog, a rat, a mouse, a horse, and a whale), or a bird (e.g., a duck or a goose).

[0158] In certain embodiments, the subject is a human, such as a human being treated or assessed for a disease, disorder, or condition in which reduced HSD17B13 expression is expected to be beneficial, as described herein; a human being at risk for a disease, disorder, or condition in which reduced HSD17B13 expression is expected to be beneficial; a human being with a disease, disorder, or condition in which reduced HSD17B13 expression is expected to be beneficial; and / or a human being receiving treatment for a disease, disorder, or condition in which reduced HSD17B13 expression is expected to be beneficial.

[0159] In one embodiment, the subject is heterozygous for the gene encoding the patatin-like phospholipase domain-containing 3 (PNPLA3) I148M variation. In another embodiment, the subject is homozygous for the gene encoding the PNPLA3 I148M variation. In one embodiment, the subject is heterozygous for the gene encoding the patatin-like phospholipase domain-containing 3 (PNPLA3) I144M variation. In another embodiment, the subject is homozygous for the gene encoding the PNPLA3 I144M variation. In one embodiment, the subject is homozygous for the gene encoding a functional HSD17B13 protein. In another embodiment, the subject is heterozygous for the gene encoding a functional HSD17B13 protein. In yet another embodiment, the subject is heterozygous for the gene encoding a functional HSD17B13 protein and the gene encoding a loss-of-function variant of HSD17B13. In another embodiment, the subject is not a carrier of the HSD17B13 rs72613567 variant, eg, HSD17B13 rs72613567:TA.

[0160] As used herein, the term "treating" or "treatment" refers to a beneficial or desired result, including, but not limited to, alleviating or reversing one or more symptoms associated with HSD17B13 gene expression and / or HSD17B13 protein production, e.g., chronic fibroinflammatory liver disease, e.g., liver inflammation, liver fibrosis, HSD17B13-associated diseases such as nonalcoholic steatohepatitis (NASH), nonalcoholic fatty liver disease (NAFLD), cirrhosis, alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), HCV-associated cirrhosis, drug-induced liver injury, hepatocellular necrosis, and / or hepatocellular carcinoma. "Treatment" can also mean prolonging survival compared to expected survival if no treatment is administered.

[0161] The term "reduce" refers to a statistically significant reduction in such level in relation to an HSD17B13-related disease. The reduction can be, for example, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more. In certain embodiments, the reduction is at least 20%. Preferably, "reduce" refers to a reduction in the level of HSD17B13 in a subject to a level that is accepted as being within the normal range for individuals without such a disorder.

[0162] As used herein, "prevention" or "preventing," when used in reference to a disease, disorder, or condition in which reduced expression of the HSD17B13 gene is expected to be beneficial, refers to a reduction in the likelihood that a subject will develop symptoms associated with such disease, disorder, or condition, e.g., symptoms of HSD17B13 gene expression, such as liver inflammation, liver fibrosis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), cirrhosis, alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), HCV-associated cirrhosis, drug-induced liver injury, hepatocellular necrosis, and / or hepatocellular carcinoma. Effective prevention is considered to be an inability of the disease, disorder, or condition to develop, or a reduction in the onset of symptoms associated with such disease, disorder, or condition (e.g., by at least about 10% on a clinically accepted scale for the disease or disorder), or a delayed (e.g., by days, weeks, months, or years) delay in symptoms (e.g., a reduction in lipid accumulation in the liver and / or an increase in lipid droplets in the liver).

[0163] As used herein, the term "HSD17B13-associated disease" refers to a disease or disorder caused by or associated with HSD17B13 gene expression or HSD17B13 protein production. The term "HSD17B13-associated disease" includes diseases, disorders, or conditions in which reducing HSD17B13 gene expression or protein activity is expected to be beneficial.

[0164] In one embodiment, the "HSD17B13-related disease" is a chronic fibroinflammatory liver disease. A "chronic fibroinflammatory liver disease" is any disease, disorder, or condition associated with chronic liver inflammation and / or fibrosis. Non-limiting examples of chronic fibroinflammatory liver diseases include, for example, liver inflammation, liver fibrosis, nonalcoholic steatohepatitis (NASH), nonalcoholic fatty liver disease (NAFLD), cirrhosis, alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), HCV-related cirrhosis, drug-induced liver injury, hepatocellular necrosis, and / or hepatocellular carcinoma.

[0165] A "therapeutically effective amount," as used herein, is intended to include the amount of an RNAi agent that, when administered to a subject having an HSD17B13-related disease, disorder, or condition, is sufficient to effectively treat the disease (e.g., by attenuating, reversing, or maintaining an existing disease or one or more symptoms of the disease). A "therapeutically effective amount" may vary depending on the RNAi agent, how the agent is administered, the disease and its severity, as well as medical history, age, weight, family history, genetic makeup, type of prior or concurrent treatment, if any, and other individual characteristics of the subject being treated.

[0166] As used herein, a "prophylactically effective amount" includes an amount of iRNA that, when administered to a subject with an HSD17B13-related disease, disorder, or condition, is sufficient to prevent or ameliorate the disease or one or more symptoms of the disease. Ameliorating a disease includes slowing the course of the disease or reducing the severity of subsequent disease. A "prophylactically effective amount" may vary depending on the iRNA, the manner of administration of the agent, the degree of risk for the disease, and the patient's medical history, age, weight, family history, genetic makeup, type of prior or concurrent treatment, if any, and other individual characteristics of the patient being treated.

[0167] A "therapeutically effective amount" or a "prophylactically effective amount" also includes that amount of an RNAi agent that produces some desired local or systemic effect at a reasonable benefit / risk ratio applicable to any treatment. The iRNAs used in the methods of the invention can be administered in amounts sufficient to produce a reasonable benefit / risk ratio applicable to such treatment.

[0168] The phrase "pharmaceutically acceptable" is used herein to refer to compounds, materials, compositions, and / or dosage forms that are within the scope of good medical judgment and are suitable for use in contact with the tissues of human and animal subjects without excessive toxicity, irritation, allergic response, or other problem or complication, and are commensurate with a reasonable benefit / risk ratio.

[0169] The phrase "pharmaceutically acceptable carrier," as used herein, refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, magnesium talc, calcium or zinc stearate, or stearic acid), or material encapsulating a vehicle involved in carrying or transporting a compound of interest from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the subject being treated. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; and (7) lubricants, such as magnesium stearate. (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) pH buffer solutions; (21) polyesters, polycarbonates, and / or polyanhydrides; (22) bulking agents, such as polypeptides and amino acids; (23) serum components, such as serum albumin, HDL, and LDL; and (24) other non-toxic, compatible substances used in pharmaceutical formulations.

[0170] The term "sample," as used herein, includes a collection of similar fluids, cells, or tissues isolated from a subject, as well as fluids, cells, or tissues present within a subject. Examples of biological fluids include blood, serum and serous fluid, plasma, cerebrospinal fluid, ocular fluid, lymph, urine, saliva, and the like. A tissue sample may include a sample from a tissue, organ, or localized region. For example, a sample may be derived from a specific organ, part of an organ, or fluids or cells within those organs. In certain embodiments, a sample may be derived from the liver (e.g., the entire liver or a specific segment of the liver or a specific type of cell within the liver, such as a hepatocyte). In some embodiments, a "sample derived from a subject" refers to blood or plasma drawn from a subject. II. iRNAs of the Invention

[0171] iRNA that inhibits the expression of target gene is described herein.In one embodiment, iRNA inhibits the expression of HSD17B13 gene.In one embodiment, iRNA agent comprises double-stranded ribonucleic acid (dsRNA) molecule for inhibiting the expression of HSD17B13 gene in cells, such as hepatocytes, for example, in subjects, such as mammals, for example, humans, with chronic fibroinflammatory liver disease, disorder or condition, for example, disease, disorder or condition associated with the accumulation and / or increase of lipid droplets in the liver and / or liver fibrosis.

[0172] dsRNA comprises an antisense strand with a complementary region that is complementary to at least a part of the mRNA that is formed under the expression of HSD17B13 gene.The complementary region is about 30 nucleotides in length or less (for example, about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19 or 18 nucleotides in length or less).When contacted with the cell that expresses target gene, iRNA inhibits the expression of target gene (for example, human, primate, non-primate or avian target gene) by at least about 10%, for example, by PCR or branched DNA (bDNA)-based method, or by protein-based method, for example, by immunofluorescence analysis using Western blotting or flow cytometry technique.

[0173] dsRNA comprises two complementary RNA strands that hybridize under the conditions that dsRNA is used to form a duplex structure.One strand (antisense strand) of dsRNA comprises a complementary region that is substantially complementary to target sequence, and generally completely complementary.Target sequence can be derived from the sequence of mRNA that is formed during the expression of HSD17B13 gene.The other strand (sense strand) comprises a region that is complementary to antisense strand, and therefore, when the two strands are combined under appropriate conditions, they hybridize and form a duplex structure.As described elsewhere herein and known in the art, the complementary sequence of dsRNA can also be contained in the self-complementary region of a single nucleic acid molecule, rather than being separate oligonucleotides.

[0174] Generally, the duplex structure is between 15 and 30 base pairs in length, e.g., 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19- 29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs in length. Ranges and lengths between the above ranges and lengths are also contemplated as part of the invention.

[0175] Similarly, the region of complementarity to the target sequence may be between 15 and 30 nucleotides in length, e.g., 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30 , 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides in length. Ranges and lengths between the above ranges and lengths are also contemplated as part of the invention.

[0176] In some embodiments, the sense and antisense strands of the dsRNA are each independently about 15 to about 30 nucleotides in length, or about 25 to about 30 nucleotides in length, e.g., each strand is independently 15 to 29, 15 to 28, 15 to 27, 15 to 26, 15 to 25, 15 to 24, 15 to 23, 15 to 22, 15 to 21, 15 to 20, 15 to 19, 15 to 18, 15 to 17, 18 to 30, 18 to 29, 18 to 28, 18 to 27, 18 to 26, 18 to 25, 18 to 24, 18 to 23, , 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides in length. In some embodiments, the dsRNA is between about 15 and about 23 nucleotides in length, or between about 25 and about 30 nucleotides in length. Generally, dsRNA is long enough to serve as a substrate for Dicer enzyme.For example, it is well known in the art that dsRNA longer than about 21-23 nucleotides can serve as a substrate for Dicer.As those skilled in the art will also recognize, the region of RNA targeted for cleavage is often a part of a larger RNA molecule, often an mRNA molecule.When relevant, the "part" of mRNA target is the continuous sequence of mRNA target that is long enough to be able to be the substrate for RNAi-directed cleavage (i.e., cleavage through RISC pathway).

[0177] The double-stranded region, for example, about 9 to 36 base pairs, for example, about 10 to 36, 11 to 36, 12 to 36, 13 to 36, 14 to 36, 15 to 36, 9 to 35, 10 to 35, 11 to 35, 12 to 35, 13 to 35, 14 to 35, 15 to 35, 9 to 34, 10 to 34, 11 to 34, 12 to 34, 13 to 34, 14 to 34, 15 to 34, 9 to 33, 10 to 33, 11 to 33, 12-33, 13-33, 14-33, 15-33, 9-32, 10-32, 11-32, 12-32, 13-32, 14-32, 15-32, 9-31, 10-31, 11-31, 12-31, 13-32, 14-31, 15-31, 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-22 Those skilled in the art will also recognize that a duplex region of 0, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs is the primary functional portion of a dsRNA. Thus, in one embodiment, an RNA molecule or complex of RNA molecules having a duplex region greater than 30 base pairs is a dsRNA to the extent that it becomes processed into a functional duplex of, for example, 15-30 base pairs that targets the desired RNA for cleavage. Thus, in one embodiment, those skilled in the art will recognize that an miRNA is a dsRNA. In another embodiment, the dsRNA is not a naturally occurring miRNA. In another embodiment, an iRNA agent useful for targeting HSD17B13 expression is not generated by cleavage of a larger dsRNA in the target cell.

[0178] The dsRNA described herein can further comprise one or more single-stranded nucleotide overhangs, for example, 1, 2, 3 or 4 nucleotides.The dsRNA with at least one nucleotide overhang can have unexpectedly superior inhibitory properties compared with their blunt-ended counterparts.The nucleotide overhang can comprise or consist of nucleotide / nucleoside analogs, including deoxynucleotides / nucleosides.The overhang can be present on the sense strand, antisense strand or any combination thereof.Furthermore, the nucleotide of the overhang can be present on the 5'-end, 3'-end or both ends of either the antisense or sense strand of dsRNA.

[0179] dsRNA can be synthesized by standard methods known in the art, discussed further below, for example, by using an automated DNA synthesizer such as those commercially available from Biosearch, Applied Biosystems, Inc.

[0180] The iRNA compounds of the present invention can be prepared using a two-step procedure.First, the individual strands of double-stranded RNA molecules are prepared separately.Then, the component strands are annealed.The individual strands of siRNA compounds can be prepared using liquid phase or solid phase organic synthesis or both.Organic synthesis has the advantage that it is easy to prepare oligonucleotide strands containing unusual nucleotides or modified nucleotides.The single-stranded oligonucleotides of the present invention can be prepared using liquid phase or solid phase organic synthesis or both.

[0181] In one embodiment, the dsRNA of the present invention comprises at least two nucleotide sequences, sense sequence and antisense sequence.Sense strand sequence is selected from the group of sequences listed in any one of Tables 2, 3, 7, 8, 10, 11 or 13, and the corresponding nucleotide sequence of the antisense strand of sense strand is selected from the group of sequences listed in any one of Tables 2, 3, 7, 8, 10, 11 or 13.In this embodiment, one of the two sequences is complementary to the other of the two sequences, and one of these sequences is substantially complementary to the sequence of the mRNA produced under the expression of HSD17B13 gene.In this embodiment, the dsRNA comprises two oligonucleotides, one oligonucleotide is listed in any one of Tables 2, 3, 7, 8, 10, 11 or 13 as sense strand (passenger strand), and the second oligonucleotide is listed in any one of Tables 2, 3, 7, 8, 10, 11 or 13 as the corresponding antisense strand (guide strand) of sense strand. In one embodiment, the substantially complementary sequences of the dsRNA are contained in separate oligonucleotides, hi another embodiment, the substantially complementary sequences of the dsRNA are contained in a single oligonucleotide.

[0182] Although the sequences in Tables 2, 3, 7, 8, 10, 11, or 13 are described as modified, unmodified, unconjugated, and / or conjugated sequences, it is understood that the RNA of the iRNA of the invention, e.g., the dsRNA of the invention, can comprise any one of the sequences set forth in any one of Tables 2, 3, 7, 8, 10, 11, or 13 unmodified, unconjugated, and / or modified and / or conjugated differently than those described herein.

[0183] Those skilled in the art are well aware that dsRNAs having a duplex structure of about 20 to 23 base pairs, for example, 21 base pairs, are recognized to be particularly effective in inducing RNA interference (Elbashir et al., (2001) EMBO J., 20: 6877-6888). However, others It has been found that shorter or longer RNA duplex structures can also be effective (Chu and Rana (2007) RNA 14: 1714-1719; Kim et al. (2005) Nat Biotech 23: 222-226). In the above embodiment, the oligonucleotides presented herein Due to the nature of sequence, dsRNA described herein can comprise at least one strand with a minimum length of 21 nucleotides.It can reasonably be expected that a shorter double strand, which is only a few nucleotides removed from one or both ends, can be similarly effective compared with the above-mentioned dsRNA.Therefore, it is intended that the dsRNA that is derived from one of the sequences presented herein and has a sequence of at least 15, 16, 17, 18, 19, 20 or more consecutive nucleotides, and its ability to inhibit the expression of HSD17B13 gene differs from the dsRNA that comprises the entire sequence by about 5, 10, 15, 20, 25 or 30% or less.

[0184] Furthermore, RNAs listed in any one of Tables 2, 3, 7, 8, 10, 11, or 13 identify sites within the HSD17B13 transcript that are susceptible to RISC-mediated cleavage. Thus, the present invention further features iRNAs that target within this site. As used herein, an iRNA is said to target within a specific site of an RNA transcript if it promotes cleavage of the transcript anywhere within the specific site. Such iRNAs generally contain at least about 15 contiguous nucleotides from one of the sequences presented herein, coupled with additional nucleotide sequences taken from regions adjacent to the selected sequence within the gene.

[0185] Target sequences are generally about 15-30 nucleotides in length, although the suitability of specific sequences within this range for directing cleavage of any given target RNA varies widely. While the various software packages and guidelines presented herein provide guidance for identifying optimal target sequences for any given gene target, an empirical approach can also be taken: a "window" or "mask" of a given size (21 nucleotides, as a non-limiting example) is placed literally or figuratively (including, for example, in silico) on the target RNA sequence to identify sequences within that size range that can serve as target sequences. Subsequent potential target sequences can be identified by successively shifting the sequence "window" one nucleotide upstream or downstream from the position of the initial target sequence until a complete set of possible sequences is identified for any given target size selected. This process, coupled with systematic synthesis and testing of identified sequences (using assays described herein or known in the art) to identify optimally functioning sequences, can identify RNA sequences that mediate the best inhibition of target gene expression when targeted with an iRNA agent. Thus, while the sequences identified herein represent effective target sequences, it is contemplated that further optimization of inhibitory efficiency can be achieved by successively "window walking" one nucleotide upstream or downstream of a given sequence to identify sequences with equal or better inhibitory properties.

[0186] Furthermore, it is contemplated that further optimization can be achieved by systematically adding or removing nucleotides from any sequence identified herein to generate longer or shorter sequences, and testing the generated sequences by moving up or down from that point through longer or shorter size windows.Again, this method for generating new candidate targets can be combined with testing the effectiveness of iRNAs based on these target sequences in inhibition assays known in the art and / or described herein, leading to further improvements in inhibition efficiency.Furthermore, such optimized sequences can be adjusted, for example, by introducing modified nucleotides described herein or known in the art, adding or changing overhangs, or other modifications known in the art and / or discussed herein, to further optimize the molecule as an expression inhibitor (e.g., increasing serum stability or circulating half-life, increasing thermostability, enhancing transmembrane delivery, targeting specific locations or cell types, increasing interaction with silencing pathway enzymes, increasing release from endosomes).

[0187] The iRNA agents described herein may contain one or more mismatches to the target sequence. In one embodiment, the iRNAs described herein contain three or fewer mismatches. If the antisense strand of an iRNA contains mismatches to the target sequence, the region of mismatch is preferably not located in the center of the complementary region. If the antisense strand of an iRNA contains mismatches to the target sequence, the mismatch is preferably limited to within the last five nucleotides from either the 5' or 3' end of the complementary region. For example, for a 23-nucleotide iRNA agent, the strand complementary to a region of the HSD17B13 gene generally does not contain any mismatches within the central 13 nucleotides. Methods described herein or known in the art can be used to determine whether an iRNA containing mismatches to the target sequence is effective in inhibiting expression of the HSD17B13 gene. Considering the effectiveness of iRNAs with mismatches in inhibiting expression of the HSD17B13 gene is important, especially when a particular complementary region of the HSD17B13 gene is known to have polymorphic sequence variation within the population. III. Modified iRNAs of the Invention

[0188] In one embodiment, the RNA, e.g., dsRNA, of an iRNA of the present invention is unmodified, e.g., does not contain chemical modifications and / or conjugations known in the art and described herein. In another embodiment, the RNA, e.g., dsRNA, of an iRNA of the present invention is chemically modified to enhance stability or other beneficial properties. In certain embodiments of the present invention, substantially all of the nucleotides of an iRNA of the present invention are modified. In other embodiments of the present invention, all of the nucleotides of an iRNA of the present invention are modified. An iRNA of the present invention in which "substantially all of the nucleotides are modified" is largely, but not entirely, modified and may contain no more than 5, 4, 3, 2, or 1 unmodified nucleotide.

[0189] In some aspects of the invention, substantially all of the nucleotides of an iRNA of the invention are modified, and the iRNA agent includes 10 or fewer nucleotides that include a 2'-fluoro modification (e.g., 9 or fewer 2'-fluoro modifications, 8 or fewer 2'-fluoro modifications, 7 or fewer 2'-fluoro modifications, 6 or fewer 2'-fluoro modifications, 5 or fewer 2'-fluoro modifications, 4 or fewer 2'-fluoro modifications, 5 or fewer 2'-fluoro modifications, 4 or fewer 2'-fluoro modifications, 3 or fewer 2'-fluoro modifications, or 2 or fewer 2'-fluoro modifications). For example, in some embodiments, the sense strand includes 4 or fewer nucleotides that include a 2'-fluoro modification (e.g., 3 or fewer 2'-fluoro modifications, or 2 or fewer 2'-fluoro modifications). In other embodiments, the antisense strand includes 6 or fewer nucleotides that include a 2'-fluoro modification (e.g., 5 or fewer 2'-fluoro modifications, 4 or fewer 2'-fluoro modifications, 4 or fewer 2'-fluoro modifications, or 2 or fewer 2'-fluoro modifications).

[0190] In other aspects of the invention, all of the nucleotides of an iRNA of the invention are modified, and the iRNA agent includes 10 or fewer nucleotides that include 2'-fluoro modifications (e.g., 9 or fewer 2'-fluoro modifications, 8 or fewer 2'-fluoro modifications, 7 or fewer 2'-fluoro modifications, 6 or fewer 2'-fluoro modifications, 5 or fewer 2'-fluoro modifications, 4 or fewer 2'-fluoro modifications, 5 or fewer 2'-fluoro modifications, 4 or fewer 2'-fluoro modifications, 3 or fewer 2'-fluoro modifications, or 2 or fewer 2'-fluoro modifications).

[0191] In one embodiment, the double-stranded RNAi agent of the present invention further comprises a 5'-phosphate or a 5'-phosphate mimic at the 5' nucleotide of the antisense strand. In another embodiment, the double-stranded RNAi agent further comprises a 5'-phosphate mimic at the 5' nucleotide of the antisense strand. In a specific embodiment, the 5'-phosphate mimic is 5'-vinyl phosphate (5'-VP).

[0192] The nucleic acids featured in the present invention are those described in "Current protocols in nucleic acid chemistry," Beaucage, SL et al. (Eds.), John Wiley & Sons, Inc., New York, NY, USA, which is hereby incorporated by reference. The iRNA compounds can be synthesized and / or modified by methods well established in the art, such as those described herein. Modifications include, for example, terminal modifications, such as 5'-end modifications (phosphorylation, conjugation, reverse linkage) or 3'-end modifications (conjugation, DNA nucleotides, reverse linkage, etc.); base modifications, such as replacement with stabilizing bases, destabilizing bases, or bases that base pair with an expanded repertoire of partners, base removal (abasic nucleotides), or conjugated bases; sugar modifications (e.g., at the 2' or 4' position) or sugar replacement; and / or backbone modifications, including modification or replacement of phosphodiester linkages. Specific examples of iRNA compounds useful in the embodiments described herein include, but are not limited to, RNAs containing modified backbones or that do not contain natural internucleoside linkages. RNAs with modified backbones include, inter alia, RNAs that do not have a phosphorus atom in the backbone. For purposes herein, and as sometimes referred to in the art, modified RNAs that do not have a phosphorus atom in the internucleoside backbone can also be considered oligonucleosides. In some embodiments, the modified iRNA has a phosphorus atom in the internucleoside backbone.

[0193] Modified RNA backbones include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkyl phosphotriesters, methyl and other alkyl phosphonates, including 3'-alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates, including 3'-amino phosphoramidates and aminoalkyl phosphoramidates, thionophosphoramidates, thionoalkyl phosphonates, thionoalkyl phosphotriesters, and boranophosphates with normal 3'-5' linkages, their 2'-5' linked analogs, and those with reverse polarity, in which adjacent pairs of nucleoside units are linked 3'-5' to 5'-3' or 2'-5' to 5'-2'. Various salts, mixed salts, and free acid forms are also included. In some embodiments of the present invention, the dsRNA agents of the present invention are in the form of free acids. In other embodiments of the present invention, the dsRNA agents of the present invention are in the form of salts. In one embodiment, the dsRNA agents of the present invention are in the form of sodium salts. In certain embodiments, when the dsRNA agent of the present invention is in the form of sodium salt, sodium ions are present in the agent as counterions to substantially all of the phosphodiester and / or phosphorothioate groups present in the agent.Agents in which substantially all of the phosphodiester and / or phosphorothioate linkages have sodium counterions include 5, 4, 3, 2, or 1 or less phosphodiester and / or phosphorothioate linkages without sodium counterions.In some embodiments, when the dsRNA agent of the present invention is in the form of sodium salt, sodium ions are present in the agent as counterions to all of the phosphodiester and / or phosphorothioate groups present in the agent.

[0194] Representative United States patents that teach the preparation of the above phosphorus-containing linkages include, but are not limited to, U.S. Patent Nos. 3,687,808; 4,469,863; 4,476,301; 5,023,243; 5,177,195; 5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,71 No. 7; No. 5,321,131; No. 5,399,676; No. 5,405,939; No. 5,453,496; No. 5,455,233; No. 5,466,677; No. 5,476 , No. 5,519,126; No. 5,536,821; No. 5,541,316; No. 5,550,111; No. 5,563,253; No. 5,571,799; No. 5, 587,361; 5,625,050; 6,028,188; 6,124,445; 6,160,109; 6,169,170; 6,172,209; No. 6,239,265; No. 6,277,603; No. 6,326,199; No. 6,346,614; No. 6,444,423; No. 6,531,590; No. 6,534,639 Nos. 6,608,035; 6,683,167; 6,858,715; 6,867,294; 6,878,805; 7,015,315; 7,041,816; 7,273,933; 7,321,029; and U.S. Patent RE39464, the entire contents of each of which are hereby incorporated by reference herein.

[0195] Modified RNA backbones that do not contain phosphorus atoms have backbones formed by short alkyl or cycloalkyl internucleoside linkages, mixed heteroatom and alkyl or cycloalkyl internucleoside linkages, or one or more short heteroatom or heterocyclic internucleoside linkages. These include morpholino linkages (formed in part from the sugar portion of the nucleoside), siloxane backbones, sulfide, sulfoxide, and sulfone backbones, formacetyl and thioformacetyl backbones, methyleneformacetyl and thioformacetyl backbones, alkene-containing backbones, sulfamic acid backbones, methyleneimino and methylenehydrazino backbones, sulfonic acid and sulfonamide backbones, amide backbones, and others with mixed N, O, S, and CH2 moieties.

[0196] Representative United States patents which teach the preparation of the above oligonucleosides include, but are not limited to, U.S. Patent Nos. 5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141; 5,235,033; 5,64,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967; Nos. 489,677; 5,541,307; 5,561,225; 5,596,086; 5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070; 5,663,312; 5,633,360; 5,677,437; and 5,677,439, the entire contents of each of which are hereby incorporated by reference herein.

[0197] In other embodiments, suitable RNA mimics for use in iRNA are contemplated, in which both the sugar and internucleoside linkages, i.e., the backbone, of the nucleotide units are replaced with novel groups. The base units for hybridization with appropriate nucleic acid target compounds are maintained. One such oligomeric compound, an RNA mimic that has been shown to have excellent hybridization properties, is called peptide nucleic acid (PNA). In PNA compounds, the sugar backbone of RNA is replaced with an amide-containing backbone, particularly an aminoethylglycine backbone. The nucleobases are retained and are directly or indirectly linked to the aza nitrogen atoms of the amide portion of the backbone. Representative U.S. patents that teach the preparation of PNA compounds include, but are not limited to, U.S. Patent Nos. 5,539,082; 5,714,331; and 5,719,262, the entire contents of each of which are hereby incorporated by reference. Additional PNA compounds suitable for use in the iRNA of the present invention are described, for example, in Nielsen et al., Science, 1991, 254, 1497-1500. It is listed.

[0198] Some embodiments featured in the present invention include oligonucleosides having the phosphorothioate backbone of the above-referenced U.S. Patent No. 5,489,677, and heteroatom backbones, specifically --CH2--NH--CH2-, --CH2--N(CH3)--O--CH2-- (known as the methylene (methylimino) or MMI backbone), --CH2--O--N(CH3)--CH2--, --CH2--N(CH3)--N(CH3)--CH2--, and --N(CH3)--CH2--CH2-- (the native phosphodiester backbone is represented by --O--P--O--CH2--) as well as RNAs having the amide backbone of the above-referenced U.S. Patent No. 5,602,240. In some embodiments, the RNAs featured herein have the morpholino backbone structure of the above-referenced U.S. Patent No. 5,034,506.

[0199] Modified RNAs may also contain one or more substituted sugar moieties. The iRNAs, e.g., dsRNAs, featured herein may include one of the following at the 2' position: OH; F; O-alkyl, S-alkyl, or N-alkyl; O-alkenyl, S-alkenyl, or N-alkenyl; O-alkynyl, S-alkynyl, or N-alkynyl; or O-alkyl-O-alkyl, where alkyl, alkenyl, and alkynyl are substituted or unsubstituted C1-C6. 10 Alkyl or C2-C 10 It can be alkenyl and alkynyl. Exemplary suitable modifications include O[(CH) n O] m CH3, O(CH2). n OCH3, O(CH2) n NH2, O(CH2) n CH3, O(CH2) n ONH2, and O(CH2) n ON[(CH2) n CH3)]2, where n and m are from 1 to about 10. In other embodiments, the dsRNA includes one of the following at the 2' position: C1 to C 10lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH, OCN, Cl, Br, CN, CF, OCF, SOCH, SOCH, ONO, NO, N, NH, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, RNA cleaving group, reporter group, intercalator, group for improving the pharmacokinetic properties of iRNA, or group for improving the pharmacodynamic properties of iRNA, and other substituents with similar properties. In some embodiments, the modification comprises 2'-methoxyethoxy (2'-O-CHCHOCH, also known as 2'-O-(2-methoxyethyl) or 2'-MOE) (Martin et al., Helv. Chim. Acta, 1995, 78:486-504), i.e., an alkoxy-alkoxy group. Another exemplary modification is the 2'-dimethylaminooxyethoxy, also known as 2'-DMAOE, i.e., O(CH2)2ON(CH3)2 group, described herein in the Examples below, and 2'-dimethylaminoethoxyethoxy (also known in the art as 2'-O-dimethylaminoethoxyethyl or 2'-DMAEOE), i.e., 2'-O-CH2-O-CH2-N(CH2)2. Further exemplary modifications include 5'-Me-2'-F nucleotides, 5'-Me-2'-OMe nucleotides, 5'-Me-2'-deoxynucleotides (both the R and S isomers of these three families); 2'-alkoxyalkyl; and 2'-NMA (N-methylacetamide).

[0200] Other modifications include 2'-methoxy (2'-OCH), 2'-aminopropoxy (2'-OCHCHCHNH), and 2'-fluoro (2'-F). Similar modifications can be made at other positions in the RNA of an iRNA, particularly the 3' position of the sugar of the 3'-terminal nucleotide or the 5' position of the 5'-terminal nucleotide in 2'-5'-linked dsRNA. An iRNA can also have a sugar mimic, such as a cyclobutyl moiety, in place of the pentofuranosyl sugar. Representative United States patents that teach the preparation of such modified sugar structures include, but are not limited to, U.S. Patent Nos. 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; Nos. 5,567,811; 5,576,427; 5,591,722; 5,597,909; 5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,658,873; 5,670,633; and 5,700,920, some of which are commonly owned with the present application. The entire contents of each of the foregoing are hereby incorporated by reference herein.

[0201] The iRNAs of the present invention may also include modifications or substitutions of nucleobases (often simply referred to in the art as "bases"). As used herein, "unmodified" or "natural" nucleobases include the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U). Modified nucleobases include other synthetic and natural nucleobases, such as 5-methylcytosine (5-me-C), 5-hydroxymethylcytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyluracil and cytosine, 6-azouracil, cytosine and thymine, 5-uracil ... uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl, and other 8-substituted adenines and guanines, 5-halo, particularly 5-bromo, 5-trifluoromethyl, and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine, and 3-deazaguanine and 3-deazaadenine. Additional nucleobases include those disclosed in U.S. Pat. No. 3,687,808 and Modified Nucleosides in Biochemistry, Biotechnology and Medicine, Herdewijn, P. ed. Wiley-VCH, 2008; Disclosed in The Concise Encyclopedia of Polymer Science and Engineering, pages 858-859, Kroschwitz, J. L, ed. John Wiley & Sons, 1990. , as disclosed by Englisch et al., (1991) Angewandte Chemie, International Edition, 30: 613, and Sanghvi, Y S., Chapter 15, dsRNA Research and Applications, pages 289-302, Crooke, ST and Lebleu, B., Eds., CRC Press, 1993. Some of these nucleobases are particularly useful for increasing the binding affinity of the oligomeric compounds featured in the present invention. These include 5-substituted pyrimidines, including 2-aminopropyladenine, 5-propynyluracil and 5-propynylcytosine, 6-azapyrimidines, and N-2, N-6 and O-6 substituted purines. 5-methylcytosine substitution has been shown to increase nucleic acid duplex stability by 0.6 to 1.2°C (Sanghvi, YS, Crooke, ST and Lebleu, B., Eds., dsRNA Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278), and even more. Specifically, it is an exemplary base substitution when combined with a 2'-O-methoxyethyl sugar modification.

[0202] Representative United States patents that teach the preparation of some of the above-mentioned modified nucleobases, as well as other modified nucleobases, include, but are not limited to, the above-mentioned U.S. Patent Nos. 3,687,808; 4,845,205; 5,130,30; 5,134,066; 5,175,273; 5,367,066; 5,432,272; 5,457,187; 5,459,255; 5,484,908; 5,502,177; 5,525,711; 5,552,540; 5,587,469; 5,594, Nos. 121, 5,596,091; 5,614,617; 5,681,941; 5,750,692; 6,015,886; 6,147,200; 6,166,197; 6,222,025; 6,235,887; 6,380,368; 6,528,640; 6,639,062; 6,617,438; 7,045,610; 7,427,672; and 7,495,088, the entire contents of each of which are hereby incorporated by reference herein.

[0203] The iRNA of the present invention may be modified to include one or more locked nucleic acids (LNAs). A locked nucleic acid is a nucleotide with a modified ribose moiety that contains an additional bridge connecting the 2' and 4' carbons. This structure effectively "locks" the ribose in a 3'-end structural conformation. The addition of locked nucleic acids to siRNA has been shown to increase siRNA stability in serum and reduce off-target effects (Elmen, J. et al., (2005) Nucleic Acids Research 33 (1): 439-447; Mook, OR. et al., (2007) Mol Canc Ther 6 (3): 833-843; Grunweller, A. et al., (2003) Nucleic Acids Research 31 (12): 3185-3193).

[0204] iRNAs of the present invention may be modified to include one or more bicyclic sugar moieties. A "bicyclic sugar" is a furanosyl ring modified by a two-atom bridge. A "bicyclic nucleoside" ("BNA") is a nucleoside having a sugar moiety that includes a bridge connecting two carbon atoms of the sugar ring, thereby forming a bicyclic ring system. In certain embodiments, the bridge connects the 4'-carbon and 2'-carbon of the sugar ring. Thus, in some embodiments, agents of the present invention may include one or more locked nucleic acids (LNAs). A locked nucleic acid is a nucleotide with a modified ribose moiety that includes an additional bridge connecting the 2'-carbon and 4'-carbon of the ribose moiety. In other words, an LNA is a nucleotide containing a bicyclic sugar moiety that includes a 4'-CH2-O-2' bridge. This structure effectively "locks" the ribose into a 3'-terminal structural conformation. The addition of locked nucleic acids to siRNA has been shown to increase siRNA stability in serum and reduce off-target effects (Elmen, J. et al., (2005) Nucleic Acids Research 33 (1): 439-447; Mook, OR. et al., (2007) Mol Canc Ther 6 (3): 833-843; Grunweller, A. et al., (2003) Nucleic Acids Research 31 (12): 3185-3193). Examples of bicyclic nucleosides for use in polynucleotides of the invention include, without limitation, nucleosides comprising a bridge between the 4'-ribosyl ring atom and the 2'-ribosyl ring atom. In certain embodiments, antisense polynucleotide agents of the invention comprise one or more bicyclic nucleosides comprising a 4'-2' bridge. Examples of such 4'-2' bridged bicyclic nucleosides include, but are not limited to, 4'-(CH2)-O-2' (LNA); 4'-(CH2)-S-2'; 4'-(CH2)2-O-2' (ENA); 4'-CH(CH3)-O-2' (also referred to as "constrained ethyl" or "cEt") and 4'-CH(CHOCH3)-O-2' (and analogs thereof; see, e.g., U.S. Pat. No. 7,399,845); 4'-C(CH3)(CH3)-O-2' (and analogs thereof; see, e.g., U.S. Pat. No. 8,272,727). 8,283); 4'-CH2-N(OCH3)-2' (and analogs thereof; see, e.g., U.S. Pat. No. 8,278,425); 4'-CH2-ON(CH3)-2' (see, e.g., U.S. Patent Publication No. 2004 / 0171570); 4'-CH2-N(R)-O-2' (wherein R is H, C1-C12 alkyl, or a protecting group) (see, e.g., U.S. Pat. No. 7,427,672); 4'-CH2-C(H)(CH3)-2' (see, e.g., Chattopadhyaya et al., J. Org. Chem., 2009, 74, 118-134). and 4'-CH2-C(=CH2)-2' (and analogs thereof; see, e.g., U.S. Pat. No. 8,278,426). The entire contents of each of the foregoing are hereby incorporated by reference herein.

[0205] Additional representative U.S. patents and publications that teach the preparation of locked nucleic acid nucleotides include, but are not limited to, the following: U.S. Patent Nos. 6,268,490; 6,525,191; 6,670,461; 6,770,748; 6,794,499; 6,998,484; 7,053,207; 7,034,133; 7,084,125; ,399,845; 7,427,672; 7,569,686; 7,741,457; 8,022,193; 8,030,467; 8,278,425; 8,278,426; 8,278,283; US2008 / 0039618; and US2009 / 0012281, the entire contents of each of which are hereby incorporated by reference herein.

[0206] Any of the foregoing bicyclic nucleosides can be prepared with one or more stereochemical sugar configurations, including, for example, α-L-ribofuranose and β-D-ribofuranose (see WO 99 / 14226).

[0207] The iRNA of the present invention may be modified to contain one or more constrained ethyl nucleotides. As used herein, a "constrained ethyl nucleotide" or "cEt" is a locked nucleic acid containing a bicyclic sugar moiety containing a 4'-CH(CH3)-0-2' bridge. In one embodiment, the constrained ethyl nucleotide is in the S conformation and is referred to herein as an "S-cEt."

[0208] The iRNA of the present invention may also contain one or more "conformationally restricted nucleotides" ("CRNs"). A CRN is a nucleotide analogue with a linker connecting the C2' and C4' carbons of ribose or the C3 and -C5' carbons of ribose. The CRN locks the ribose ring into a stable conformation, increasing hybridization affinity to mRNA. The linker is long enough to place the oxygen in an optimal position for stability and affinity, resulting in minimal distortion of the ribose ring.

[0209] Representative publications that teach the preparation of some of the above CRNs include, but are not limited to, U.S. Patent Publication No. 2013 / 0190383; and PCT Publication No. WO2013 / 036868, the entire contents of each of which are hereby incorporated by reference herein.

[0210] In some embodiments, the iRNA of the present invention comprises one or more monomers that are UNA (unlocked nucleic acid) nucleotides. UNAs are unlocked acyclic nucleic acids in which one of the sugar linkages has been removed, thereby forming an unlocked "sugar" residue. In one example, UNAs also encompass monomers in which the C1'-C4' bond (i.e., the carbon-oxygen-carbon covalent bond between the C1' and C4' carbons) has been removed. In another example, the C2'-C3' bond (i.e., the carbon-carbon covalent bond between the C2' and C3' carbons) of the sugar has been removed (Nuc. Acids Symp. Series, 52, 133-134 (2008) and Fluiter et al., Mol. Biosyst., 2009, which are hereby incorporated by reference). 10, 1039).

[0211] Representative U.S. publications teaching the preparation of UNAs include, but are not limited to, U.S. Patent No. 8,314,227; and U.S. Patent Publication Nos. 2013 / 0096289; 2013 / 0011922; and 2011 / 0313020, the entire contents of each of which are hereby incorporated by reference herein.

[0212] Potentially stabilizing modifications to the ends of RNA molecules include N-(acetylaminocaproyl)-4-hydroxyprolinol (Hyp-C6-NHAc), N-(caproyl-4-hydroxyprolinol (Hyp-C6), N-(acetyl-4-hydroxyprolinol (Hyp-NHAc), thymidine-2'-0-deoxythymidine (ether), N-(aminocaproyl)-4-hydroxyprolinol (Hyp-C6-amino), 2-docosanoyl-uridine-3''-phosphate, inverted base dT (idT), and others. Disclosure of this modification can be found in PCT Publication No. WO2011 / 005861.

[0213] Other modifications of the iRNA of the present invention include a 5' phosphate or 5' phosphate mimic, such as a 5' terminal phosphate or phosphate mimic, on the antisense strand of the RNAi agent. Suitable phosphate mimics are disclosed, for example, in U.S. Patent Publication No. 2012 / 0157511, the entire contents of which are incorporated herein by reference.

[0214] In certain specific embodiments, the RNAi agent of the present invention is an agent that inhibits expression of the HSD17B13 gene selected from the group of agents listed in any one of Tables 2, 3, 7, 8, 10, 11, or 13. Any of these agents may further comprise a ligand. A. Modified iRNAs Containing Motifs of the Invention

[0215] In certain embodiments of the invention, double-stranded RNAi agents of the invention include agents with chemical modifications, for example, as disclosed in WO2013 / 075035, filed November 16, 2012, the entire contents of which are incorporated herein by reference.

[0216] Thus, the present invention provides a double-stranded RNAi agent capable of inhibiting the expression of a target gene (i.e., the HSD17B13 gene) in vivo. The RNAi agent comprises a sense strand and an antisense strand. Each strand of the RNAi agent can range from 12 to 30 nucleotides in length. For example, each strand can be 14 to 30 nucleotides in length, 17 to 30 nucleotides in length, 25 to 30 nucleotides in length, 27 to 30 nucleotides in length, 17 to 23 nucleotides in length, 17 to 21 nucleotides in length, 17 to 19 nucleotides in length, 19 to 25 nucleotides in length, 19 to 23 nucleotides in length, 19 to 21 nucleotides in length, 21 to 25 nucleotides in length, or 21 to 23 nucleotides in length. In one embodiment, the sense strand is 21 nucleotides in length. In one embodiment, the antisense strand is 23 nucleotides in length.

[0217] The sense strand and antisense strand generally form a double-stranded RNA ("dsRNA"), also referred to herein as an "RNAi agent." The duplex region of the RNAi agent can be 12 to 30 nucleotide pairs in length. For example, the duplex region can be 14 to 30 nucleotide pairs in length, 17 to 30 nucleotide pairs in length, 27 to 30 nucleotide pairs in length, 17 to 23 nucleotide pairs in length, 17 to 21 nucleotide pairs in length, 17 to 19 nucleotide pairs in length, 19 to 25 nucleotide pairs in length, 19 to 23 nucleotide pairs in length, 19 to 21 nucleotide pairs in length, 21 to 25 nucleotide pairs in length, or 21 to 23 nucleotide pairs in length. In another example, the duplex region is selected from the group consisting of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, and 27 nucleotides in length.

[0218] In one embodiment, the RNAi agent may contain one or more overhanging regions and / or capping groups at the 3'-end, 5'-end, or both ends of one or both strands. The overhangs may be 1 to 6 nucleotides in length, e.g., 2 to 6 nucleotides in length, 1 to 5 nucleotides in length, 2 to 5 nucleotides in length, 1 to 4 nucleotides in length, 2 to 4 nucleotides in length, 1 to 3 nucleotides in length, 2 to 3 nucleotides in length, or 1 to 2 nucleotides in length. The overhangs may be the result of one strand being longer than the other, or may be the result of two strands of the same length being offset. The overhangs may form mismatches with the target mRNA, or may be complementary to the targeted gene sequence, or may be another sequence. The first and second strands may be joined by additional bases, for example, to form a hairpin, or by other non-basic linkers.

[0219] In one embodiment, the nucleotides in the overhang region of the RNAi agent can each independently be modified or unmodified nucleotides, including, but not limited to, 2'-sugar modified nucleotides, such as 2-F, 2'-O-methyl, thymidine (T), 2'-O-methoxyethyl-5-methyluridine (Teo), 2'-O-methoxyethyl adenosine (Aeo), 2'-O-methoxyethyl-5-methylcytidine (m5Ceo), and any combination thereof. For example, TT can be an overhang sequence at either end of either strand. The overhang can form a mismatch with the target mRNA, or can be complementary to the targeted gene sequence, or can be a different sequence.

[0220] The 5'-overhang or 3'-overhang of the sense strand, antisense strand, or both strands of the RNAi agent may be phosphorylated.In some embodiments, the overhang region comprises two nucleotides with phosphorothioate between them, wherein the two nucleotides may be the same or different.In one embodiment, the overhang is present at the 3'-end of the sense strand, antisense strand, or both strands.In one embodiment, the 3'-overhang is present in the antisense strand.In one embodiment, the 3'-overhang is present in the sense strand.

[0221] RNAi agents can contain only a single overhang, thereby enhancing the interference activity of RNAi without affecting the overall stability of RNAi.For example, the single-stranded overhang can be located at the 3'-end of the sense strand, or alternatively, at the 3'-end of the antisense strand.RNAi can also have a blunt end located at the 5'-end of the antisense strand (or the 3'-end of the sense strand), or vice versa.Generally, the antisense strand of RNAi has a nucleotide overhang at its 3'-end, and its 5'-end is blunt.Without wishing to be bound by theory, the asymmetric blunt end at the 5'-end of the antisense strand and the 3'-end overhang of the antisense strand are favorable for guide strand loading into RISC process.

[0222] In one embodiment, the RNAi agent is a 19-nucleotide double-ended bluntmer, wherein the sense strand contains at least one motif with three 2'-F modifications in three consecutive nucleotides at positions 7, 8, and 9 from the 5' end, and the antisense strand contains at least one motif with three 2'-O-methyl modifications in three consecutive nucleotides at positions 11, 12, and 13 from the 5' end.

[0223] In another embodiment, the RNAi agent is 20 nucleotides in length and blunt at both ends, and the sense strand contains at least one motif with three 2'-F modifications in three consecutive nucleotides at positions 8, 9, and 10 from the 5' end, and the antisense strand contains at least one motif with three 2'-O-methyl modifications in three consecutive nucleotides at positions 11, 12, and 13 from the 5' end.

[0224] In yet another embodiment, the RNAi agent is 21 nucleotides in length and blunt at both ends, and the sense strand contains at least one motif with three 2'-F modifications in three consecutive nucleotides at positions 9, 10, and 11 from the 5' end, and the antisense strand contains at least one motif with three 2'-O-methyl modifications in three consecutive nucleotides at positions 11, 12, and 13 from the 5' end.

[0225] In one embodiment, the RNAi agent comprises a 21-nucleotide sense strand and a 23-nucleotide antisense strand, wherein the sense strand contains at least one motif with three 2'-F modifications in three consecutive nucleotides at positions 9, 10, and 11 from the 5' end, and the antisense strand contains at least one motif with three 2'-O-methyl modifications in three consecutive nucleotides at positions 11, 12, and 13 from the 5' end, and one end of the RNAi agent is blunt and the other end comprises a two-nucleotide overhang, preferably at the 3' end of the antisense strand.

[0226] When a two-nucleotide overhang is at the 3'-end of the antisense strand, there may be two phosphorothioate internucleotide linkages between the three terminal nucleotides, where two of the three nucleotides are overhanging nucleotides, and the third nucleotide is the paired nucleotide next to the overhanging nucleotide.In one embodiment, the RNAi agent further has two phosphorothioate internucleotide linkages between the three terminal nucleotides at both the 5'-end of the sense strand and the 5'-end of the antisense strand.In one embodiment, all nucleotides in the sense strand and antisense strand of the RNAi agent, including the nucleotides that are part of a motif, are modified nucleotides.In one embodiment, each residue is independently modified with 2'-O-methyl or 3'-fluoro, for example, as an alternating motif.Optionally, the RNAi agent further comprises a ligand (preferably GalNAc3).

[0227] In one embodiment, the RNAi agent comprises a sense strand and an antisense strand, wherein the sense strand is 25 to 30 nucleotide residues in length, starting from the 5'-terminal nucleotide (position 1), and comprises at least 8 ribonucleotides at positions 1 to 23 of the first strand; the antisense strand is 36 to 66 nucleotide residues in length, starting from the 3'-terminal nucleotide, and comprises at least 8 ribonucleotides at positions 1 to 23 of the sense strand that pair with the sense strand to form a duplex; at least the 3'-terminal nucleotide of the antisense strand is unpaired with the sense strand, and up to 6 consecutive 3'-terminal nucleotides are unpaired with the sense strand, thereby forming a 3' single-stranded overhang of 1 to 6 nucleotides; and the 5'-end of the antisense strand is unpaired with 10 to 30 consecutive nucleotides that are not paired with the sense strand. the sense strand contains at least one ribonucleotide with three 2'-F modifications in three consecutive nucleotides, thereby forming a single-stranded 5' overhang of 10 to 30 nucleotides; when the sense and antisense strands are aligned for maximum complementarity, at least the 5'- and 3'-terminal nucleotides of the sense strand base pair with nucleotides of the antisense strand, thereby forming a substantially duplex region between the sense and antisense strands; and the antisense strand is sufficiently complementary to the target RNA along at least 19 ribonucleotides of the antisense strand so that target gene expression is reduced when the double-stranded nucleic acid is introduced into a mammalian cell; and the sense strand contains at least one motif with three 2'-F modifications in three consecutive nucleotides, at least one of the motifs being at or near the cleavage site. The antisense strand contains at least one motif with three 2'-O-methyl modifications in three consecutive nucleotides at or near the cleavage site.

[0228] In one embodiment, the RNAi agent comprises a sense and an antisense strand, the RNAi agent comprising a first strand at least 25 and at most 29 nucleotides in length and a second strand at most 30 nucleotides in length, the second strand having at least one motif with three 2'-O-methyl modifications at three consecutive nucleotides, positions 11, 12, and 13, from the 5' end; the 3' end of the first strand and the 5' end of the second strand form a blunt end; the second strand is 1-4 nucleotides longer at its 3' end than the first strand; the duplex region is at least 25 nucleotides in length; the second strand is sufficiently complementary to a target mRNA along the length of at least 19 nucleotides of the second strand such that target gene expression is reduced when the RNAi agent is introduced into a mammalian cell; and cleavage of the RNAi agent by Dicer preferentially yields siRNA comprising the 3' end of the second strand, thereby reducing target gene expression in the mammal. Optionally, the RNAi agent further comprises a ligand.

[0229] In one embodiment, the sense strand of the RNAi agent contains at least one motif with three identical modifications to three consecutive nucleotides, one of the motifs being at the cleavage site of the sense strand.

[0230] In one embodiment, the antisense strand of the RNAi agent may also contain at least one motif having three identical modifications to three consecutive nucleotides, one of the motifs being at or near the cleavage site on the antisense strand.

[0231] For RNAi agents having a duplex region 17-23 nucleotides long, the cleavage site of the antisense strand is generally near positions 10, 11, and 12 from the 5' end. Thus, motifs with three identical modifications can be present at positions 9, 10, and 11; 10, 11, and 12; 11, 12, and 13; 12, 13, and 14; or 13, 14, and 15 of the antisense strand, with counting starting from the first nucleotide from the 5' end of the antisense strand, or from the first paired nucleotide within the duplex region from the 5' end of the antisense strand. The cleavage site of the antisense strand from the 5' end can also vary depending on the length of the duplex region of the RNAi.

[0232] The sense strand of RNAi agent can contain at least one motif with three identical modifications for three consecutive nucleotides at the breakpoint of strand; antisense strand can have at least one motif with three identical modifications for three consecutive nucleotides at or near the breakpoint of strand.When sense strand and antisense strand form dsRNA duplex, sense strand and antisense strand can be aligned so that one motif of three nucleotides in sense strand and one motif of three nucleotides in antisense strand have at least one nucleotide overlap, that is, at least one of the three nucleotides of the motif in sense strand and at least one of the three nucleotides of the motif in antisense strand form base pairs.Alternatively, at least two nucleotides can be overlapped, or all three nucleotides can be overlapped.

[0233] In one embodiment, the sense strand of an RNAi agent may contain more than one motif with three identical modifications to three consecutive nucleotides. The first motif may be at or near the cleavage site of the strand, and the other motif may be a wing modification. The term "wing modification" as used herein refers to a motif that is present in another part of the strand, separated from a motif at or near the cleavage site of the same strand. The wing modification may be adjacent to the first motif or separated by at least one or more nucleotides. When the motifs are immediately adjacent to each other, the chemical properties of the motifs are different from each other, and when the motifs are separated by one or more nucleotides, the chemical properties may be the same or different. Two or more wing modifications may be present. For example, when two wing modifications are present, each wing modification may be present at one end of the first motif at or near the cleavage site, or on both sides of the lead motif.

[0234] Like the sense strand, the antisense strand of an RNAi agent can contain more than one motif with three identical modifications to three consecutive nucleotides, at least one of which is at or near the site of strand cleavage. The antisense strand can also contain one or more wing modifications in the same alignment as the wing modifications that may be present in the sense strand.

[0235] In one embodiment, wing modifications on the sense or antisense strand of an RNAi agent generally do not include the first one or two terminal nucleotides at the 3' end, 5' end, or both ends of the strand.

[0236] In another embodiment, wing modifications on the sense or antisense strand of an RNAi agent generally do not include the first one or two paired nucleotides in the duplex region at the 3' end, 5' end, or both ends of the strand.

[0237] When the sense and antisense strands of an RNAi agent each contain at least one wing modification, the wing modifications can be located at the same end of the duplex region and can have an overlap of 1, 2, or 3 nucleotides.

[0238] When the sense and antisense strands of an RNAi agent each contain at least two wing modifications, the sense and antisense strands can be aligned such that the two modifications from each strand are located at one end of the duplex region and overlap by one, two, or three nucleotides; the two modifications from each strand are located at the other end of the duplex region and overlap by one, two, or three nucleotides; or the two modifications on one strand are located on either side of the lead motif and overlap by one, two, or three nucleotides within the duplex region.

[0239] In one embodiment, every nucleotide in the sense and antisense strands of an RNAi agent can be modified, including nucleotides that are part of a motif. Each nucleotide can be modified with the same or different modifications, which can include one or more changes to one or both of the free phosphate oxygens and / or one or more of the linked phosphate oxygens; changes to the composition of the ribose sugar, for example, the 2' hydroxyl of the ribose sugar; wholesale replacement of the phosphate moiety with a "dephospho" linker; modifications or replacements of naturally occurring bases; and replacements or modifications of the ribose-phosphate backbone.

[0240] Because nucleic acids are polymers of subunits, many modifications occur at positions that are repeated within nucleic acids, such as base modifications, or phosphate moieties, or unlinked Os in phosphate moieties. In some cases, modifications occur at all target positions within nucleic acids, but in many cases, this is not the case. For example, modifications can occur only at the 3' or 5' end, or only in the terminal region, for example, at the terminal nucleotide position of the chain, or only in the last 2, 3, 4, 5, or 10 nucleotides. Modifications can occur in double-stranded regions, single-stranded regions, or both. Modifications can occur only in the double-stranded region of RNA, or only in the single-stranded region of RNA. For example, phosphorothioate modifications at unlinked O positions can occur only at one or both ends, or only in the terminal region, for example, at the terminal nucleotide position of the chain, or only in the last 2, 3, 4, 5, or 10 nucleotides, or in double-stranded and single-stranded regions, especially at the ends. One or more 5' ends can be phosphorylated.

[0241] For example, it may be possible to enhance stability, include specific bases in the overhang, or include modified nucleotides or nucleotide surrogates in the single-stranded overhang, e.g., the 5' or 3' overhang, or both. For example, it may be desirable to include purine nucleotides in the overhang. In some embodiments, all or some of the bases in the 3' or 5' overhang may be modified, e.g., with the modifications described herein. Modifications can include, for example, the use of modifications at the 2' position of the ribose sugar, e.g., deoxyribonucleotides in place of the ribosugar of the nucleobase, 2'-deoxy-2'-fluoro (2'-F) or 2'-O-methyl modifications, and modifications at the phosphate group, e.g., phosphorothioate modifications, using modifications known in the art. The overhang need not be homologous to the target sequence.

[0242] In one embodiment, each residue of sense strand and antisense strand is independently modified with LNA, CRN, cET, UNA, HNA, CeNA, 2'-methoxyethyl, 2'-O-methyl, 2'-O-allyl, 2'-C-allyl, 2'-deoxy, 2'-hydroxyl or 2'-fluoro.Strands can contain more than one modification.In one embodiment, each residue of sense strand and antisense strand is independently modified with 2'-O-methyl or 2'-fluoro.

[0243] Generally, at least two different modifications are present in the sense and antisense strands. These two modifications can be 2'-O-methyl or 2'-fluoro modifications, or others.

[0244] In one embodiment, N a and / or N b includes an alternating pattern of modifications. The term "alternating motif," as used herein, refers to a motif having one or more modifications, with each modification occurring at alternating nucleotides on one strand. Alternating nucleotides can refer to one every other nucleotide or one every third nucleotide, or a similar pattern. For example, if A, B, and C each represent one type of modification to a nucleotide, the alternating motif could be "ABABABABABAB...," "AABBAABBAABB...," "AABAABAABAAB...," "AAABAAABAAAB...," "AAABBBAAABBB...," or "ABCABCABCABC...," etc.

[0245] The types of modifications contained within the alternating motif can be the same or different. For example, if A, B, C, and D each represent one type of modification to a nucleotide, the alternation pattern, i.e., the modifications to every other nucleotide, can be the same, but each of the sense or antisense strands can be selected from several modification possibilities within the alternating motif, such as "ABABAB...", "ACACAC...", "BDBDBD..." or "CDCDCD...".

[0246] In one embodiment, the RNAi agent of the present invention comprises a modification pattern of the alternating motif in the sense strand that is shifted relative to the modification pattern of the alternating motif in the antisense strand. The shift can be such that the modified group of the nucleotide in the sense strand corresponds to the differently modified group of the nucleotide in the antisense strand, or vice versa. For example, when the sense strand and the antisense strand are paired in a dsRNA duplex, the alternating motif in the sense strand can begin with "ABABAB" from 5' to 3' of the strand, and the alternating motif in the antisense strand can begin with "BABABA" from 5' to 3' of the strand, within the duplex region. As another example, the alternating motif in the sense strand can begin with "AABBAABB" from 5' to 3' of the strand, and the alternating motif in the antisense strand can begin with "BBAABBAA" from 5' to 3' of the strand, within the duplex region, thus resulting in a complete or partial shift in the modification pattern between the sense strand and the antisense strand.

[0247] In one embodiment, the RNAi agent comprises a pattern of alternating 2'-O-methyl and 2'-F modifications initially in the sense strand, with a shift relative to the pattern of alternating 2'-O-methyl and 2'-F modifications initially in the antisense strand, i.e., 2'-O-methyl modified nucleotides in the sense strand bases are paired with 2'-F modified nucleotides in the antisense strand, and vice versa. Position 1 of the sense strand may start with a 2'-F modification, and position 1 of the antisense strand may start with a 2'-O-methyl modification.

[0248] By introducing one or more motifs that have three identical modifications for three consecutive nucleotides into sense strand and / or antisense strand, the original modification pattern that exists in sense strand and / or antisense strand is interrupted.By introducing one or more motifs that have three identical modifications for three consecutive nucleotides into sense strand and / or antisense strand, the interruption of the modification pattern of this sense strand and / or antisense strand by introducing one or more motifs that have three identical modifications for three consecutive nucleotides into sense strand and / or antisense strand surprisingly enhances the gene silencing activity for target gene.

[0249] In one embodiment, when a motif having three identical modifications to three consecutive nucleotides is introduced into either strand, the modification of the nucleotide next to the motif is a different modification than the modification of the motif. For example, the portion of the sequence containing the motif is denoted by "...N a YYYN b ...", where "Y" represents a modification of the motif with three identical modifications on three consecutive nucleotides, and "N a " and "N b " represents a modification to the nucleotide next to the motif "YYY" that is different from the modification Y, and N a and N b may be the same with modifications or different modifications, or, if wing modifications are present, N a and / or N b may or may not be present.

[0250] The RNAi agent may further comprise at least one phosphorothioate or methylphosphonate internucleotide linkage.The phosphorothioate or methylphosphonate internucleotide linkage modification may be present at any position of the chain with respect to any nucleotide of the sense strand or antisense strand or both strands.For example, the internucleotide linkage modification may be present at every nucleotide of the sense strand and / or antisense strand; each internucleotide linkage modification may be present in an alternating pattern in the sense strand and / or antisense strand; or the sense strand or antisense strand may contain both internucleotide linkage modifications in an alternating pattern.The alternating pattern of the internucleotide linkage modification in the sense strand may be the same or different from that of the antisense strand, and the alternating pattern of the internucleotide linkage modification in the sense strand may have a shift relative to the alternating pattern of the internucleotide linkage modification in the antisense strand.In one embodiment, the double-stranded RNAi agent comprises 6-8 phosphorothioate internucleotide linkages. In one embodiment, the antisense strand comprises two phosphorothioate internucleotide linkages at the 5' end and two phosphorothioate internucleotide linkages at the 3' end, and the sense strand comprises at least two phosphorothioate internucleotide linkages at either the 5' end or the 3' end.

[0251] In one embodiment, the RNAi comprises a phosphorothioate or methylphosphonate internucleotide linkage modification in the overhanging region. For example, the overhanging region can contain two nucleotides with a phosphorothioate or methylphosphonate internucleotide linkage between the two nucleotides. The internucleotide linkage modification can also be created so that the overhanging nucleotide is linked to the terminal paired nucleotide in the double-stranded region. For example, at least two, three, four, or all of the overhanging nucleotides can be linked through phosphorothioate or methylphosphonate internucleotide linkages, and optionally, there can be an additional phosphorothioate or methylphosphonate internucleotide linkage connecting the overhanging nucleotide to the paired nucleotide next to the overhanging nucleotide. For example, there can be at least two phosphorothioate internucleotide linkages between the terminal three nucleotides, where two of the three nucleotides are overhanging nucleotides and the third nucleotide is the paired nucleotide next to the overhanging nucleotide. These terminal three nucleotides can be at the 3'-end of the antisense strand, the 3'-end of the sense strand, the 5'-end of the antisense strand, and / or the 5'-end of the antisense strand.

[0252] In one embodiment, the two-nucleotide overhang is at the 3'-end of the antisense strand, and two phosphorothioate internucleotide linkages are present between the terminal three nucleotides, where two of the three nucleotides are overhanging nucleotides, and the third nucleotide is the paired nucleotide next to the overhanging nucleotide.Optionally, the RNAi agent can further have two phosphorothioate internucleotide linkages between the terminal three nucleotides at both the 5'-end of the sense strand and the 5'-end of the antisense strand.

[0253] In one embodiment, the RNAi agent contains mismatches with the target, mismatches within the duplex, or a combination thereof. The mismatches can be present in the overhang region or the duplex region. Base pairs can be ranked based on their tendency to promote dissociation or melting (e.g., based on the free energy of association or dissociation of a particular pairing; the simplest approach is to examine each pair individually, but proximity or similar analysis can also be used). In terms of promoting dissociation, A:U is preferred over G:C, G:U is preferred over G:C, and I:C is preferred over G:C (I=inosine). Mismatches, such as non-standard or non-standard pairings (as described elsewhere herein), are preferred over standard (A:T, A:U, G:C) pairings, and pairings involving universal bases are preferred over standard pairings.

[0254] In one embodiment, the RNAi agent comprises at least one of the first 1, 2, 3, 4, or 5 base pairs within the duplex region, from the 5' end of the antisense strand, independently selected from the group of A:U, G:U, I:C, and mismatch pairs, e.g., non-standard or non-standard pairings or pairings comprising universal bases, to promote dissociation of the antisense strand at the 5' end of the duplex.

[0255] In one embodiment, the nucleotide at position 1 in the double-stranded region from the 5' end of the antisense strand is selected from the group consisting of A, dA, dU, U and dT.Alternatively, at least one of the first 1, 2 or 3 base pairs in the double-stranded region from the 5' end of the antisense strand is an AU base pair.For example, the first base pair in the double-stranded region from the 5' end of the antisense strand is an AU base pair.

[0256] In another embodiment, the 3'-terminal nucleotide of the sense strand is deoxythymine (dT). In another embodiment, the 3'-terminal nucleotide of the antisense strand is deoxythymine (dT). In one embodiment, there is a short sequence of deoxythymine nucleotides, for example, two dT nucleotides, at the 3'-end of the sense and / or antisense strand. In one embodiment, the sense strand sequence has formula (I): 5'n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3'(I) (In the formula, i and j are each independently 0 or 1; p and q are each independently 0 to 6; each N a independently represent oligonucleotide sequences containing 0 to 25 modified nucleotides, each sequence containing at least two differently modified nucleotides; each N b independently represent an oligonucleotide sequence containing 0 to 10 modified nucleotides; each n p and n q independently represent overhanging nucleotides; Nb and Y do not have the same modification; XXX, YYY and ZZZ each independently represent one motif with three identical modifications on three consecutive nucleotides. It is preferred that all of YYY are 2'-F modified nucleotides.

[0257] In one embodiment, N a and / or N b includes alternating pattern modifications.

[0258] In one embodiment, the YYY motif is located at or near the cleavage site of the sense strand. For example, if the RNAi agent has a duplex region of 17 to 23 nucleotides in length, the YYY motif can be located at or near the cleavage site of the sense strand (e.g., at positions 6, 7, 8, 7, 8, 9, 8, 9, 10, 9, 10, 11, 10, 11, 12, or 11, 12, 13), and counting begins from the first nucleotide from the 5' end; or optionally, counting begins from the first paired nucleotide in the duplex region from the 5' end.

[0259] In one embodiment, i is 1 and j is 0, or i is 0 and j is 1, or i and j are both 1. Thus, the sense strand can be represented by the following formula: 5'n p -N a -YYY-N b -ZZZ-N a -n q 3'(Ib); 5'n p -N a -XXX-N b -YYY-N a -n q 3'(Ic); or 5'n p -N a -XXX-N b -YYY-N b -ZZZ-N a -n q 3'(Id).

[0260] When the sense strand is represented by formula (Ib), N b represents an oligonucleotide sequence containing 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0 modified nucleotides. a can independently represent an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0261] When the sense strand is represented by formula (Ic), N brepresents an oligonucleotide sequence containing 0 to 10, 0 to 7, 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0 modified nucleotides. a can independently represent an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0262] When the sense strand is represented by formula (Id), each N b represents an oligonucleotide sequence containing, independently, 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0 modified nucleotides. b is preferably 0, 1, 2, 3, 4, 5 or 6. a can independently represent an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0263] Each of X, Y and Z may be the same or different from the others.

[0264] In other embodiments, i is 0, j is 0, and the sense strand has the formula: 5'n p -N a -YYY-N a -n q 3'(Ia) It can be expressed as:

[0265] When the sense strand is represented by formula (Ia), each N a can independently represent an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0266] In one embodiment, the antisense strand sequence of the RNAi has the formula (II): 5'n q’ -N a '-(Z'Z'Z') k -N b '-Y'Y'Y'-N b '-(X'X'X') l -N' a -n p'3'(II) (In the formula, k and l are each independently 0 or 1; p' and q' are each independently 0 to 6; each N a ' independently represent oligonucleotide sequences containing 0 to 25 modified nucleotides, each sequence containing at least two differently modified nucleotides; each N b ' independently represents an oligonucleotide sequence containing 0 to 10 modified nucleotides; each n p ' and n q ' independently represent an overhanging nucleotide; N b ' and Y' do not have the same modification; X'X'X', Y'Y'Y' and Z'Z'Z' each independently represent a motif with three identical modifications on three consecutive nucleotides. It can be expressed as:

[0267] In one embodiment, N a ' and / or N b ' includes alternating pattern modifications.

[0268] The Y'Y'Y' motif is present at or near the cleavage site of the antisense strand. For example, if the RNAi agent has a duplex region 17 to 23 nucleotides long, the Y'Y'Y' motif can be present at positions 9, 10, 11; 10, 11, 12; 11, 12, 13; 12, 13, 14; or 13, 14, 15 of the antisense strand, counting from the first nucleotide from the 5' end; or, optionally, counting from the first paired nucleotide in the duplex region from the 5' end. Preferably, the Y'Y'Y' motif is present at positions 11, 12, or 13.

[0269] In one embodiment, the Y'Y'Y' motif is all 2'-OMe modified nucleotides.

[0270] In one embodiment, k is 1 and l is 0, or k is 0 and l is 1, or k and l are both 1.

[0271] Thus, the antisense strand can be represented by the following formula: 5'n q’ -N a '-Z'Z'Z'-N b '-Y'Y'Y'-N a '-n p’ 3'(IIb); 5'n q’ -N a '-Y'Y'Y'-N b '-X'X'X'-n p’ 3'(IIc); or 5'n q’ -N a '-Z'Z'Z'-N b '-Y'Y'Y'-N b '-X'X'X'-N a '-n p’ 3'(IId).

[0272] When the antisense strand is represented by formula (IIb), N b ’ represents an oligonucleotide sequence containing 0 to 10, 0 to 7, 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0 modified nucleotides. a ' independently represent an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0273] When the antisense strand is represented by formula (IIc), N b ' represents an oligonucleotide sequence containing 0 to 10, 0 to 7, 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0 modified nucleotides. a ' independently represent an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0274] When the antisense strand is represented by formula (IId), each N b Each N' independently represents an oligonucleotide sequence containing 0 to 10, 0 to 7, 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0 modified nucleotides. a N' independently represents an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides. b is preferably 0, 1, 2, 3, 4, 5 or 6.

[0275] In other embodiments, k is 0, l is 0, and the antisense strand has the formula: 5'n p’ -N a’ -Y'Y'Y'-N a’ -n q’ 3'(Ia) It can be expressed as:

[0276] When the antisense strand is represented by formula (IIa), each N a ' independently represent an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0277] X', Y' and Z' may be the same or different from each other.

[0278] Each nucleotide of sense strand and antisense strand can be independently modified with LNA, CRN, UNA, cEt, HNA, CeNA, 2'-methoxyethyl, 2'-O-methyl, 2'-O-allyl, 2'-C-allyl, 2'-hydroxyl or 2'-fluoro.For example, each nucleotide of sense strand and antisense strand is independently modified with 2'-O-methyl or 2'-fluoro.Each X, Y, Z, X', Y' and Z' can specifically represent 2'-O-methyl modification or 2'-fluoro modification.

[0279] In one embodiment, the sense strand of the RNAi agent may contain a YYY motif occurring at positions 9, 10, and 11 of the strand when the duplex region is 21 nt, counting starting from the first nucleotide from the 5' end, or optionally counting starting from the first paired nucleotide in the duplex region from the 5' end; Y represents a 2'-F modification. The sense strand may further contain a XXX motif or a ZZZ motif as a wing modification at the opposite end of the duplex region; XXX and ZZZ each independently represent a 2'-OMe modification or a 2'-F modification.

[0280] In one embodiment, the antisense strand may contain a Y'Y'Y' motif at positions 11, 12, and 13 of the strand, counting starting from the first nucleotide from the 5' end, or optionally counting starting from the first paired nucleotide in the duplex region from the 5' end; Y' represents a 2'-O-methyl modification. The antisense strand may further contain an X'X'X' motif or a Z'Z'Z' motif as a wing modification at the opposite end of the duplex region; X'X'X' and Z'Z'Z' each independently represent a 2'-OMe or 2'-F modification.

[0281] The sense strand represented by any one of the above formulas (Ia), (Ib), (Ic), and (Id) forms a duplex with the antisense strand represented by any one of formulas (IIa), (IIb), (IIc), and (IId), respectively.

[0282] Thus, an RNAi agent for use in the methods of the invention may comprise a sense strand and an antisense strand, each strand having 14 to 30 nucleotides, and the RNAi duplex may have the formula (III): Sense: 5'np-Na-(XXX)i-Nb-YYY-Nb-(ZZZ)j-Na-nq3' Antisense: 3'np'-Na'-(X'X'X')k-Nb'-Y'Y'Y'-N b'-(Z'Z'Z')l-Na'-nq'5'(III) (In the formula, i, j, k, and l are each independently 0 or 1; p, p', q, and q' are each independently 0 to 6; each Na and Na' independently represents an oligonucleotide sequence containing 0 to 25 modified nucleotides, each sequence containing at least two differently modified nucleotides; each Nb and Nb' independently represents an oligonucleotide sequence containing 0 to 10 modified nucleotides; each np', np, nq', and nq, each of which may be present or absent, independently represents an overhanging nucleotide; XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent one motif with three identical modifications to three consecutive nucleotides. is expressed by

[0283] In one embodiment, i is 0 and j is 0; or i is 1 and j is 0; or i is 0 and j is 1; or i and j are both 0; or i and j are both 1. In another embodiment, k is 0 and l is 0; or k is 1 and l is 0; k is 0 and l is 1; or k and l are both 0; or k and l are both 1.

[0284] Exemplary combinations of sense and antisense strands that form RNAi duplexes include the following formulas: 5'np-Na-YYY-Na-nq3' 3'np'-Na'-Y'Y'Y'-Na'nq'5'(IIIa) 5'np-Na-YYY-Nb-ZZZ-Na-nq3' 3'np'-Na'-Y'Y'Y'-Nb'-Z'Z'Z'-Na'nq'5'(IIIb) 5'np-Na-XXX-Nb-YYY-Na-nq3' 3'np'-Na'-X'X'X'-Nb'-Y'Y'Y'-Na'-nq'5'(IIIc) 5'np-Na-XXX-Nb-YYY-Nb-ZZZ-Na-nq3' 3'np'-Na'-X'X'X'-Nb'-Y'Y'Y'-Nb'-Z'Z'Z'-Na-nq'5'(IIId)

[0285] When the RNAi agent is represented by formula (IIIa), each Na independently represents an oligonucleotide sequence comprising 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0286] When an RNAi agent is represented by formula (IIIb), each Nb independently represents an oligonucleotide sequence containing 1 to 10, 1 to 7, 1 to 5, or 1 to 4 modified nucleotides, and each Na independently represents an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0287] When an RNAi agent is represented as formula (IIIc), each Nb, Nb' independently represents an oligonucleotide sequence containing 0 to 10, 0 to 7, 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0 modified nucleotides, and each Na independently represents an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides.

[0288] When an RNAi agent is represented as formula (IIId), each Nb, Nb' independently represents an oligonucleotide sequence containing 0 to 10, 0 to 7, 0 to 10, 0 to 7, 0 to 5, 0 to 4, 0 to 2, or 0 modified nucleotides. Each Na, Na' independently represents an oligonucleotide sequence containing 2 to 20, 2 to 15, or 2 to 10 modified nucleotides. Na, Na', Nb, and Nb' each independently comprise an alternating pattern of modifications.

[0289] In formulae (III), (IIIa), (IIIb), (IIIc), and (IIId), X, Y, and Z may be the same as or different from each other.

[0290] When an RNAi agent is represented by formula (III), (IIIa), (IIIb), (IIIc), and (IIId), at least one of the Y nucleotides may be base-paired to one of the Y' nucleotides, alternatively, at least two of the Y nucleotides are base-paired to a corresponding Y' nucleotide; or alternatively, all three Y nucleotides are base-paired to a corresponding Y' nucleotide.

[0291] When the RNAi agent is represented by formula (IIIb) or (IIId), at least one of the Z nucleotides can be base-paired to one of the Z' nucleotides, or at least two of the Z nucleotides can be base-paired to a corresponding Z' nucleotide; or all three of the Z nucleotides can be base-paired to a corresponding Z' nucleotide.

[0292] When an RNAi agent is represented as formula (IIIc) or (IIId), at least one of the X nucleotides can be base-paired to one of the X' nucleotides, or at least two of the X nucleotides can be base-paired to a corresponding X' nucleotide; or all three X nucleotides can be base-paired to a corresponding X' nucleotide.

[0293] In one embodiment, the modification to a Y nucleotide is different from the modification to a Y' nucleotide, the modification to a Z nucleotide is different from the modification to a Z' nucleotide, and / or the modification to an X nucleotide is different from the modification to an X' nucleotide.

[0294] In one embodiment, when the RNAi agent is represented by formula (IIId), the Na modification is a 2'-O-methyl or 2'-fluoro modification. In another embodiment, when the RNAi agent is represented by formula (IIId), the Na modification is a 2'-O-methyl or 2'-fluoro modification, np'>0, and at least one np' is linked to an adjacent nucleotide by a phosphorothioate linkage. In yet another embodiment, when the RNAi agent is represented by formula (IIId), the Na modification is a 2'-O-methyl or 2'-fluoro modification, np'>0, and at least one np' is linked to an adjacent nucleotide by a phosphorothioate linkage, and the sense strand is conjugated to one or more GalNAc derivatives attached through a bivalent or trivalent branched linker (see below). In another embodiment, when the RNAi agent is represented by formula (IIId), the Na modification is a 2'-O-methyl or 2'-fluoro modification, np'>0, and at least one np' is linked to an adjacent nucleotide by a phosphorothioate linkage, and the sense strand comprises at least one phosphorothioate linkage, and the sense strand is conjugated to one or more GalNAc derivatives attached through a divalent or trivalent branched linker.

[0295] In one embodiment, when the RNAi agent is represented by formula (IIIa), the Na modification is a 2'-O-methyl or 2'-fluoro modification, np'>0, and at least one np' is linked to an adjacent nucleotide by a phosphorothioate linkage, the sense strand comprises at least one phosphorothioate linkage, and the sense strand is conjugated to one or more GalNAc derivatives attached through a divalent or trivalent branched linker.

[0296] In one embodiment, the RNAi agent is a multimer containing at least two duplexes represented by formula (III), (IIIa), (IIIb), (IIIc), and (IIId), and the duplexes are connected by a linker. The linker can be cleavable or non-cleavable. Optionally, the multimer further comprises a ligand. Each duplex can target the same gene or two different genes; or each duplex can target the same gene at two different target sites.

[0297] In one embodiment, the RNAi agent is a multimer containing three, four, five, six or more duplexes represented by formula (III), (IIIa), (IIIb), (IIIc), and (IIId), wherein the duplexes are connected by a linker. The linker can be cleavable or non-cleavable. Optionally, the multimer further comprises a ligand. Each duplex can target the same gene or two different genes; or each duplex can target the same gene at two different target sites.

[0298] In one embodiment, two RNAi agents represented by formula (III), (IIIa), (IIIb), (IIIc), and (IIId) are linked to each other at the 5' end, and one or both of the 3' ends are optionally conjugated to a ligand. Each of the agents can target the same gene or two different genes; or each of the agents can target the same gene at two different target sites.

[0299] In certain embodiments, the RNAi agent of the present invention may contain a small number of nucleotides containing a 2'-fluoro modification, for example, 10 or fewer nucleotides with a 2'-fluoro modification. For example, the RNAi agent may contain 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0 nucleotides with a 2'-fluoro modification. In certain embodiments, the RNAi agent of the present invention contains 10 nucleotides with a 2'-fluoro modification in the sense strand, for example, 4 nucleotides with a 2'-fluoro modification, and 6 nucleotides with a 2'-fluoro modification in the antisense strand. In another specific embodiment, the RNAi agent of the present invention contains 6 nucleotides with a 2'-fluoro modification in the sense strand, for example, 4 nucleotides with a 2'-fluoro modification, and 2 nucleotides with a 2'-fluoro modification in the antisense strand.

[0300] In other embodiments, the RNAi agent of the present invention may contain a very small number of nucleotides containing 2'-fluoro modification, for example, may contain two or less nucleotides containing 2'-fluoro modification.For example, the RNAi agent may contain two, one, or zero nucleotides with 2'-fluoro modification.In certain embodiments, the RNAi agent may contain two nucleotides with 2'-fluoro modification, for example, zero nucleotides with 2'-fluoro modification in the sense strand and two nucleotides with 2'-fluoro modification in the antisense strand.

[0301] Various publications describe the multimeric RNAi agent that can be used in the method of the present invention.Such publications include WO2007 / 091269, US Patent No. 7858769, WO2010 / 141511, WO2007 / 117686, WO2009 / 014887 and WO2011 / 031520, each of whose entire contents is hereby incorporated by reference into this specification.

[0302] As described in more detail below, RNAi agents containing one or more carbohydrate moieties conjugated to the RNAi agent can optimize one or more properties of the RNAi agent. Often, the carbohydrate moiety is attached to a modified subunit of the RNAi agent. For example, the ribose sugar of one or more ribonucleotide subunits of a dsRNA agent can be replaced with another moiety, such as a non-carbohydrate (preferably cyclic) carrier, to which a carbohydrate ligand is attached. Ribonucleotide subunits in which the ribose sugar of the subunit has been so replaced are referred to herein as ribose-replacement modified subunits (RRMS). Cyclic carriers can be carbocyclic ring systems, i.e., those in which all ring atoms are carbon atoms, or heterocyclic ring systems, i.e., those in which one or more ring atoms are heteroatoms, such as nitrogen, oxygen, or sulfur. Cyclic carriers can be monocyclic ring systems or can contain two or more rings, e.g., fused rings. Cyclic carriers can be fully saturated ring systems or can contain one or more double bonds.

[0303] Ligands can be attached to polynucleotides via carriers. The carriers include (i) at least one "backbone attachment point," preferably two "backbone attachment points," and (ii) at least one "tethering attachment point." As used herein, "backbone attachment point" refers to a functional group, e.g., a hydroxyl group, or generally a bond available and suitable for incorporating the carrier into a backbone, e.g., the phosphate backbone of a ribonucleic acid, or a modified phosphate backbone, e.g., a sulfur-containing backbone. "Tethering attachment point" (TAP) refers, in some embodiments, to a ring atom, e.g., a carbon atom or heteroatom (different from the atom providing the backbone attachment point), of a cyclic carrier to which a selected moiety is attached. This moiety can be, for example, a carbohydrate, e.g., a monosaccharide, disaccharide, trisaccharide, tetrasaccharide, oligosaccharide, or polysaccharide. Optionally, the selected moiety is connected to the cyclic carrier via an intervening tether. Thus, cyclic carriers often contain a functional group, such as an amino group, or generally provide a bond suitable for incorporating or tethering another chemical entity, such as a ligand, to the constituent ring.

[0304] The RNAi agent can be conjugated to the ligand via a carrier, which may be a cyclic group or an acyclic group. The cyclic group is preferably selected from pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolane, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuryl, and decalin, and the acyclic group is preferably selected from a serinol skeleton or a diethanolamine skeleton.

[0305] In another embodiment of the invention, the iRNA agent comprises a sense strand and an antisense strand, each strand having between 14 and 40 nucleotides. The RNAi agent has the formula (L): [ka] It can be expressed as:

[0306] In Formula (L), B1, B2, B3, B1', B2', B3', and B4' are each independently a nucleotide containing a modification selected from the group consisting of 2'-O-alkyl, 2'-substituted alkoxy, 2'-substituted alkyl, 2'-halo, ENA, and BNA / LNA. In certain embodiments, B1, B2, B3, B1', B2', B3', and B4' each contain a 2'-OMe modification. In certain embodiments, B1, B2, B3, B1', B2', B3', and B4' each contain a 2'-OMe or 2'-F modification. In certain embodiments, at least one of B1, B2, B3, B1', B2', B3', and B4' contains a 2'-ON-methylacetamide (2'-O-NMA) modification.

[0307] C1 is a heat-destabilizing nucleotide located opposite the seed region of the antisense strand (i.e., positions 2 to 8 of the 5' end of the antisense strand). For example, C1 is located in the sense strand at a position that pairs with nucleotides 2 to 8 of the 5' end of the antisense strand. In one example, C1 is located at position 15 from the 5' end of the sense strand. The C1 nucleotide has a heat-destabilizing modification, which can include an abasic modification; a mismatch with the opposite nucleotide in the duplex; and a sugar modification such as a 2'-deoxy modification or an acyclic nucleotide, e.g., an unlocked nucleic acid (UNA) or a glycerol nucleic acid (GNA). In certain embodiments, C1 is: i) a mismatch with the opposite nucleotide in the antisense strand; ii) [ka] an abasic modification selected from the group consisting of: [ka] wherein B is a modified or unmodified nucleobase and R 1 and R 2are independently H, halogen, OR3, or alkyl; and R3 is H, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, or sugar. In certain embodiments, the heat-destabilizing modification of C1 is a mismatch selected from the group consisting of G:G, G:A, G:U, G:T, A:A, A:C, C:C, C:U, C:T, U:U, T:T, and U:T; optionally, at least one nucleobase of the mismatch pair is a 2'-deoxynucleobase. In one example, the heat-destabilizing modification of C1 is GNA or [ka] is.

[0308] T1, T1', T2', and T3' each independently represent a nucleotide containing a modification that provides the nucleotide with steric bulk less than or equal to that of a 2'-OMe modification. Steric bulk refers to the sum of the steric effects of the modifications. Methods for determining the steric effect of a nucleotide modification are known to those skilled in the art. The modification may be at the 2' position of the ribose sugar of the nucleotide, or a modification to a non-ribose nucleotide, an acyclic nucleotide, or a nucleotide backbone similar or equivalent to the 2' position of the ribose sugar, providing the nucleotide with steric bulk less than or equal to that of a 2'-OMe modification. For example, T1, T1', T2', and T3' are each independently selected from DNA, RNA, LNA, 2'-F, and 2'-F-5'-methyl. In certain embodiments, T1 is DNA. In certain embodiments, T1' is DNA, RNA, or LNA. In certain embodiments, T2' is DNA or RNA. In certain embodiments, T3' is DNA or RNA.

[0309] n 1 , n 3 , and q 1 are independently 4 to 15 nucleotides in length.

[0310] n 5 , q 3 , and q 7 are independently 1 to 6 nucleotides in length.

[0311] n 4 , q 2 , and q 6 are independently 1 to 3 nucleotides in length; or 4 is 0.

[0312] q 5 are independently 0 to 10 nucleotides in length.

[0313] n 2 and q 4 are independently 0 to 3 nucleotides in length.

[0314] Or, n 4 is 0 to 3 nucleotides in length.

[0315] In certain embodiments, n 4 can be 0. In one example, n 4 is 0 and q 2 and q 6 is 1. In another example, n 4 is 0 and q 2 and q 6 is 1, with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[0316] In certain embodiments, n 4 , q 2 , and q 6 are each 1.

[0317] In certain embodiments, n 2 , n4 , q 2 , q 4 , and q 6 are each 1.

[0318] In certain embodiments, when the sense strand is 19-22 nucleotides in length, C1 is at positions 14-17 of the 5' end of the sense strand, and n 4 is 1. In certain embodiments, C1 is at position 15 of the 5' end of the sense strand.

[0319] In certain embodiments, T3' begins at position 2 from the 5' end of the antisense strand. In one example, T3' is at position 2 from the 5' end of the antisense strand, and q 6 is equal to 1.

[0320] In certain embodiments, T1' begins at position 14 from the 5' end of the antisense strand. In one example, T1' is at position 14 from the 5' end of the antisense strand, and 2 is equal to 1.

[0321] In an exemplary embodiment, T3' begins at position 2 from the 5' end of the antisense strand, and T1' begins at position 14 from the 5' end of the antisense strand. In one example, T3' begins at position 2 from the 5' end of the antisense strand, and q 6 is equal to 1, T1' starts at position 14 from the 5' end of the antisense strand, and q 2 is equal to 1.

[0322] In certain embodiments, T1' and T3' are separated by 11 nucleotides (ie, not counting the T1' and T3' nucleotides).

[0323] In certain embodiments, T1' is at position 14 from the 5' end of the antisense strand. In one example, T1' is at position 14 from the 5' end of the antisense strand, and 2 is equal to 1 and the modification is at the 2' position or at a non-ribose, acyclic, or backbone position that provides less steric bulk than 2'-OMe ribose.

[0324] In certain embodiments, T3' is at position 2 from the 5' end of the antisense strand. In one example, T3' is at position 2 from the 5' end of the antisense strand, and q 6 is equal to 1 and the modification is at the 2' position or at a non-ribose, acyclic, or backbone position that provides less or equal steric bulk than 2'-OMe ribose.

[0325] In certain embodiments, T1 is at the cleavage site of the sense strand. In one example, when the sense strand is 19-22 nucleotides long, T1 is at position 11 from the 5' end of the sense strand, and n 2 is 1. In an exemplary embodiment, when the sense strand is 19-22 nucleotides in length, T1 is at the cleavage site of the sense strand, position 11 from the 5' end of the sense strand, and n 2 is 1.

[0326] In certain embodiments, T2' begins at position 6 from the 5' end of the antisense strand. In one example, T2' is at positions 6-10 from the 5' end of the antisense strand, and 4 is 1.

[0327] In an exemplary embodiment, T1 is at the cleavage site of the sense strand, e.g., at position 11 from the 5' end of the sense strand when the sense strand is 19-22 nucleotides in length, and 2 is 1; T1' is at position 14 from the 5' end of the antisense strand, and q 2 is equal to 1, the modification to T1' is at the 2' position of the ribose sugar or at a non-ribose, acyclic, or backbone position that provides less steric bulk than 2'-OMe ribose; T2' is at positions 6-10 from the 5' end of the antisense strand, and q 4 is 1; and T3' is at the second position from the 5' end of the antisense strand, and q 6 is equal to 1, and the modification to T3' is at the 2' position or at a non-ribose, acyclic, or backbone position that provides less or equal steric bulk than 2'-OMe ribose.

[0328] In certain embodiments, T2' begins at position 8 from the 5' end of the antisense strand. In one example, T2' begins at position 8 from the 5' end of the antisense strand and 4 is 2.

[0329] In certain embodiments, T2' begins at position 9 from the 5' end of the antisense strand. In one example, T2' is at position 9 from the 5' end of the antisense strand, and 4 is 1.

[0330] In certain embodiments, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 1, B3' is 2'-OMe or 2'-F, and q 5 is 6, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[0331] In certain embodiments, n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 1, B3' is 2'-OMe or 2'-F, and q 5is 6, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[0332] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1.

[0333] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[0334] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 6, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 7, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1.

[0335] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 6, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1is 7, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[0336] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 1, B3' is 2'-OMe or 2'-F, and q 5 is 6, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1.

[0337] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 1, B3' is 2'-OMe or 2'-F, and q 5 is 6, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[0338] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 5, T2' is 2'-F, and q 4 is 1, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; optionally with at least two additional TTs at the 3' end of the antisense strand.

[0339] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 5, T2' is 2'-F, and q 4 is 1, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; optionally with at least two additional TTs at the 3' end of the antisense strand; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[0340] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1.

[0341] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 (counting from the 5' end) of the sense strand, and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end).

[0342] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1.

[0343] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[0344] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1.

[0345] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[0346] The RNAi agent may contain a phosphorus-containing group at the 5'-end of the sense or antisense strand, such as 5'-phosphate (5'-P), 5'-phosphorothioate (5'-PS), 5'-phosphorodithioate (5'-PS2), 5'-vinylphosphonate (5'-VP), 5'-methylphosphonate (MePhos), or 5'-deoxy-5'-C-malonyl. [ka] When the 5'-terminal phosphorus-containing group is a 5'-terminal vinyl phosphonate (5'-VP), the 5'-VP may be a 5'-E-VP isomer (i.e., trans-vinyl phosphate, [ka] ), 5'-Z-VP isomer (i.e., cis-vinyl phosphate, [ka] ) or a mixture thereof.

[0347] In certain embodiments, the RNAi agent comprises a phosphorus-containing group at the 5'-end of the sense strand. In certain embodiments, the RNAi agent comprises a phosphorus-containing group at the 5'-end of the antisense strand.

[0348] In certain embodiments, the RNAi agent comprises a 5'-P. In certain embodiments, the RNAi agent comprises a 5'-P on the antisense strand.

[0349] In certain embodiments, the RNAi agent comprises a 5'-PS. In certain embodiments, the RNAi agent comprises a 5'-PS on the antisense strand.

[0350] In certain embodiments, the RNAi agent comprises a 5'-VP. In certain embodiments, the RNAi agent comprises a 5'-VP in the antisense strand. In certain embodiments, the RNAi agent comprises a 5'-E-VP in the antisense strand. In certain embodiments, the RNAi agent comprises a 5'-Z-VP in the antisense strand.

[0351] In certain embodiments, the RNAi agent comprises a 5'-PS2. In certain embodiments, the RNAi agent comprises a 5'-PS2 on the antisense strand.

[0352] In certain embodiments, the RNAi agent comprises a 5'-PS2. In certain embodiments, the RNAi agent comprises 5'-deoxy-5'-C-malonyl in the antisense strand.

[0353] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1. The RNAi agent also contains a 5'-PS.

[0354] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6is 1, B4' is 2'-OMe, and q 7 is 1. The RNAi agent also contains a 5'-P.

[0355] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1. The RNAi agent also includes a 5'-VP. The 5'-VP can be a 5'-E-VP, a 5'-Z-VP, or a combination thereof.

[0356] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6is 1, B4' is 2'-OMe, and q 7 is 1. The RNAi agent also includes a 5'-PS2.

[0357] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1. RNAi agents also include 5'-deoxy-5'-C-malonyl.

[0358] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18-23. The RNAi agent also includes a 5'-P.

[0359] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18-23. The RNAi agent also includes a 5'-PS.

[0360] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also includes a 5'-VP. The 5'-VP can be a 5'-E-VP, a 5'-Z-VP, or a combination thereof.

[0361] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7is 1, with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18-23. The RNAi agent also includes a 5'-PS2.

[0362] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18-23. The RNAi agent also includes 5'-deoxy-5'-C-malonyl.

[0363] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1. The RNAi agent also contains a 5'-P.

[0364] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1. The dsRNA agent also contains a 5'-PS.

[0365] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1. The RNAi agent also includes a 5'-VP. The 5'-VP can be a 5'-E-VP, a 5'-Z-VP, or a combination thereof.

[0366] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1. The RNAi agent also includes a 5'-PS2.

[0367] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1. RNAi agents also include 5'-deoxy-5'-C-malonyl.

[0368] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 (counting from the 5' end) of the sense strand, and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end). The RNAi agent also includes a 5'-P.

[0369] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 (counting from the 5' end) of the sense strand, and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end). The RNAi agent also includes a 5'-PS.

[0370] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 (counting from the 5' end) of the sense strand, and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end). The RNAi agent also includes a 5'-VP. The 5'-VP can be a 5'-E-VP, a 5'-Z-VP, or a combination thereof.

[0371] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 (counting from the 5' end) of the sense strand, and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end). The RNAi agent also includes a 5'-PS2.

[0372] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 (counting from the 5' end) of the sense strand, and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5' end). The RNAi agent also includes 5'-deoxy-5'-C-malonyl.

[0373] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1. The RNAi agent also contains a 5'-P.

[0374] In certain embodiments, B1 is 2'-OMe or 2'-F, and n1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1. The RNAi agent also contains a 5'-PS.

[0375] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1. The RNAi agent also includes a 5'-VP. The 5'-VP can be a 5'-E-VP, a 5'-Z-VP, or a combination thereof.

[0376] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1. dsRNA The RNA agent also includes a 5'-PS2.

[0377] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1. RNAi agents also include 5'-deoxy-5'-C-malonyl.

[0378] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18-23. The RNAi agent also includes a 5'-P.

[0379] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18-23. The RNAi agent also includes a 5'-PS.

[0380] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also includes a 5'-VP. The 5'-VP can be a 5'-E-VP, a 5'-Z-VP, or a combination thereof.

[0381] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1, with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18-23. The RNAi agent also includes a 5'-PS2.

[0382] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18-23. The RNAi agent also includes 5'-deoxy-5'-C-malonyl.

[0383] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1. The RNAi agent also contains a 5'-P.

[0384] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1. The RNAi agent also contains a 5'-PS.

[0385] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1. The RNAi agent also includes a 5'-VP. The 5'-VP can be a 5'-E-VP, a 5'-Z-VP, or a combination thereof.

[0386] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1. The RNAi agent also includes a 5'-PS2.

[0387] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1. RNAi agents also include 5'-deoxy-5'-C-malonyl.

[0388] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18-23. The RNAi agent also includes a 5'-P.

[0389] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18-23. The RNAi agent also includes a 5'-PS.

[0390] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also includes a 5'-VP. The 5'-VP can be a 5'-E-VP, a 5'-Z-VP, or a combination thereof.

[0391] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7is 1, with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18-23. The RNAi agent also includes a 5'-PS2.

[0392] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18-23. The RNAi agent also includes 5'-deoxy-5'-C-malonyl.

[0393] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also includes a 5'-P and a targeting ligand. In certain embodiments, the 5'-P is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0394] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also includes a 5'-PS and a targeting ligand. In certain embodiments, the 5'-PS is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0395] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1, with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand) and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18 to 23. The RNAi agent also includes a 5'-VP (e.g., 5'-E-VP, 5'-Z-VP, or a combination thereof), and a targeting ligand.

[0396] In certain embodiments, the 5'-VP is at the 5' end of the antisense strand and the targeting ligand is at the 3' end of the sense strand.

[0397] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also includes a 5'-PS2 and a targeting ligand. In certain embodiments, the 5'-PS2 is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0398] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18 to 23. The RNAi agent also includes 5'-deoxy-5'-C-malonyl and a targeting ligand. In certain embodiments, the 5'-deoxy-5'-C-malonyl is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0399] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 (counting from the 5' end) of the sense strand, and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end). The RNAi agent also includes a 5'-P and a targeting ligand. In certain embodiments, the 5'-P is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0400] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 (counting from the 5' end) of the sense strand, and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end). The RNAi agent also includes a 5'-PS and a targeting ligand. In certain embodiments, the 5'-PS is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0401] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 (counting from the 5' end) of the sense strand, and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end). The RNAi agent also includes a 5'-VP (e.g., 5'-E-VP, 5'-Z-VP, or a combination thereof) and a targeting ligand. In certain embodiments, the 5'-VP is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0402] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 (counting from the 5' end) of the sense strand, and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end). The RNAi agent also includes a 5'-PS2 and a targeting ligand. In certain embodiments, the 5'-PS2 is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0403] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-OMe, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 (counting from the 5' end) of the sense strand, and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end). The RNAi agent also includes 5'-deoxy-5'-C-malonyl and a targeting ligand. In certain embodiments, the 5'-deoxy-5'-C-malonyl is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0404] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also includes a 5'-P and a targeting ligand. In certain embodiments, the 5'-P is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0405] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also includes a 5'-PS and a targeting ligand. In certain embodiments, the 5'-PS is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0406] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18 to 23. The RNAi agent also includes a 5'-VP (e.g., 5'-E-VP, 5'-Z-VP, or a combination thereof) and a targeting ligand. In certain embodiments, the 5'-VP is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0407] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also includes a 5'-PS2 and a targeting ligand. In certain embodiments, the 5'-PS2 is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0408] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4, T2' is 2'-F, and q 4 is 2, B3' is 2'-OMe or 2'-F, and q 5 is 5, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18 to 23. The RNAi agent also includes 5'-deoxy-5'-C-malonyl and a targeting ligand. In certain embodiments, the 5'-deoxy-5'-C-malonyl is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0409] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also includes a 5'-P and a targeting ligand. In certain embodiments, the 5'-P is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0410] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also includes a 5'-PS and a targeting ligand. In certain embodiments, the 5'-PS is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0411] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18 to 23. The RNAi agent also includes a 5'-VP (e.g., 5'-E-VP, 5'-Z-VP, or a combination thereof) and a targeting ligand. In certain embodiments, the 5'-VP is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0412] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4 is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18 to 23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also includes a 5'-PS2 and a targeting ligand. In certain embodiments, the 5'-PS2 is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0413] In certain embodiments, B1 is 2'-OMe or 2'-F, and n 1 is 8, T1 is 2'F, and n 2 is 3, B2 is 2'-OMe, and n 3 is 7, and n 4 is 0, B3 is 2'-OMe, and n 5 is 3, B1' is 2'-OMe or 2'-F, and q 1 is 9, T1' is 2'-F, and q 2 is 1, B2' is 2'-OMe or 2'-F, and q 3 is 4 and q 4is 0, B3' is 2'-OMe or 2'-F, and q 5 is 7, T3' is 2'-F, and q 6 is 1, B4' is 2'-F, and q 7 is 1; with two phosphorothioate internucleotide linkage modifications within positions 1 to 5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 of the antisense strand (counting from the 5' end of the antisense strand) and two phosphorothioate internucleotide linkage modifications within positions 18 to 23. The RNAi agent also includes 5'-deoxy-5'-C-malonyl and a targeting ligand. In certain embodiments, the 5'-deoxy-5'-C-malonyl is at the 5' end of the antisense strand, and the targeting ligand is at the 3' end of the sense strand.

[0414] In certain embodiments, the RNAi agents of the invention are (a) (i) 21 nucleotides in length; (ii) an ASGPR ligand attached to its 3' end, the ASGPR ligand comprising three GalNAc derivatives attached through a trivalent branched linker; and (iii) 2'-F modifications at positions 1, 3, 5, 7, 9-11, 13, 17, 19, and 21, and 2'-OMe modifications at positions 2, 4, 6, 8, 12, 14-16, 18, and 20 (counting from the 5' end). a sense strand having: (b) (i) 23 nucleotides in length; (ii) 2'-OMe modifications at positions 1, 3, 5, 9, 11-13, 15, 17, 19, 21, and 23, and 2'F modifications at positions 2, 4, 6-8, 10, 14, 16, 18, 20, and 22 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotides 21 and 22 and between nucleotides 22 and 23 (counting from the 5' end) and an antisense strand having Including, The dsRNA agent has a two-nucleotide overhang on the 3' end of the antisense strand and a blunt end on the 5' end of the antisense strand.

[0415] In another particular embodiment, the RNAi agent of the invention is (a) (i) 21 nucleotides in length; (ii) an ASGPR ligand attached at its 3' end, the ASGPR ligand comprising three GalNAc derivatives attached through a trivalent branched linker; (iii) 2'-F modifications at positions 1, 3, 5, 7, 9-11, 13, 15, 17, 19, and 21, and 2'-OMe modifications at positions 2, 4, 6, 8, 12, 14, 16, 18, and 20 (counting from the 5' end); and (iv) phosphorothioate internucleotide linkages between nucleotides 1 and 2 and between nucleotides 2 and 3 (counting from the 5' end) and a sense strand having (b) (i) 23 nucleotides in length; (ii) 2'-OMe modifications at positions 1, 3, 5, 7, 9, 11-13, 15, 17, 19, and 21-23, and 2'F modifications at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotides 1 and 2, between nucleotides 2 and 3, between nucleotides 21 and 22, and between nucleotides 22 and 23 (counting from the 5' end) and an antisense strand having Including, The RNAi agent has a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[0416] In another particular embodiment, the RNAi agent of the invention is (a) (i) 21 nucleotides in length; (ii) an ASGPR ligand attached at its 3' end, the ASGPR ligand comprising three GalNAc derivatives attached through a trivalent branched linker; (iii) 2'-OMe modifications at positions 1 to 6, 8, 10, and 12 to 21, 2'-F modifications at positions 7 and 9, and a deoxynucleotide (e.g., dT) at position 11 (counting from the 5' end); and (iv) phosphorothioate internucleotide linkages between nucleotides 1 and 2 and between nucleotides 2 and 3 (counting from the 5' end) and a sense strand having (b) (i) 23 nucleotides in length; (ii) 2'-OMe modifications at positions 1, 3, 7, 9, 11, 13, 15, 17, and 19-23, and 2'-F modifications at positions 2, 4-6, 8, 10, 12, 14, 16, and 18 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotides 1 and 2, between nucleotides 2 and 3, between nucleotides 21 and 22, and between nucleotides 22 and 23 (counting from the 5' end) and an antisense strand having Including, The RNAi agent has a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[0417] In another particular embodiment, the RNAi agent of the invention is (a) (i) 21 nucleotides in length; (ii) an ASGPR ligand attached at its 3' end, the ASGPR ligand comprising three GalNAc derivatives attached through a trivalent branched linker; (iii) 2'-OMe modifications at positions 1 to 6, 8, 10, 12, 14, and 16 to 21, and 2'-F modifications at positions 7, 9, 11, 13, and 15; and (iv) phosphorothioate internucleotide linkages between nucleotides 1 and 2 and between nucleotides 2 and 3 (counting from the 5' end) and a sense strand having (b) (i) 23 nucleotides in length; (ii) 2'-OMe modifications at positions 1, 5, 7, 9, 11, 13, 15, 17, 19, and 21-23, and 2'-F modifications at positions 2-4, 6, 8, 10, 12, 14, 16, 18, and 20 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotides 1 and 2, between nucleotides 2 and 3, between nucleotides 21 and 22, and between nucleotides 22 and 23 (counting from the 5' end) and an antisense strand having Including, The RNAi agent has a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[0418] In another particular embodiment, the RNAi agent of the invention is (a) (i) 21 nucleotides in length; (ii) an ASGPR ligand attached at its 3' end, the ASGPR ligand comprising three GalNAc derivatives attached through a trivalent branched linker; (iii) 2'-OMe modifications at positions 1 to 9 and 12 to 21, and 2'-F modifications at positions 10 and 11; and (iv) phosphorothioate internucleotide linkages between nucleotides 1 and 2 and between nucleotides 2 and 3 (counting from the 5' end) and a sense strand having (b) (i) 23 nucleotides in length; (ii) 2'-OMe modifications at positions 1, 3, 5, 7, 9, 11-13, 15, 17, 19, and 21-23, and 2'-F modifications at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotides 1 and 2, between nucleotides 2 and 3, between nucleotides 21 and 22, and between nucleotides 22 and 23 (counting from the 5' end) and an antisense strand having Including, The RNAi agent has a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[0419] In another particular embodiment, the RNAi agent of the invention is (a) (i) 21 nucleotides in length; (ii) an ASGPR ligand attached at its 3' end, the ASGPR ligand comprising three GalNAc derivatives attached through a trivalent branched linker; (iii) 2'-F modifications at positions 1, 3, 5, 7, 9-11, and 13, and 2'-OMe modifications at positions 2, 4, 6, 8, 12, and 14-21; and (iv) phosphorothioate internucleotide linkages between nucleotides 1 and 2 and between nucleotides 2 and 3 (counting from the 5' end) and a sense strand having (b) (i) 23 nucleotides in length; (ii) 2'-OMe modifications at positions 1, 3, 5-7, 9, 11-13, 15, 17-19, and 21-23, and 2'-F modifications at positions 2, 4, 8, 10, 14, 16, and 20 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotides 1 and 2, between nucleotides 2 and 3, between nucleotides 21 and 22, and between nucleotides 22 and 23 (counting from the 5' end) and an antisense strand having Including, The RNAi agent has a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[0420] In another particular embodiment, the RNAi agent of the invention is (a) (i) 21 nucleotides in length; (ii) an ASGPR ligand attached at its 3' end, the ASGPR ligand comprising three GalNAc derivatives attached through a trivalent branched linker; (iii) 2'-OMe modifications at positions 1, 2, 4, 6, 8, 12, 14, 15, 17, and 19-21, and 2'-F modifications at positions 3, 5, 7, 9-11, 13, 16, and 18; and (iv) phosphorothioate internucleotide linkages between nucleotides 1 and 2 and between nucleotides 2 and 3 (counting from the 5' end) and a sense strand having (b) (i) 25 nucleotides in length; (ii) 2'-OMe modifications at positions 1, 4, 6, 7, 9, 11-13, 15, 17, and 19-23, 2'-F modifications at positions 2, 3, 5, 8, 10, 14, 16, and 18, and deoxynucleotides (e.g., dT) at positions 24 and 25 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotides 1 and 2, between nucleotides 2 and 3, between nucleotides 21 and 22, and between nucleotides 22 and 23 (counting from the 5' end) and an antisense strand having Including, The RNAi agent has a 4-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[0421] In another particular embodiment, the RNAi agent of the invention is (a) (i) 21 nucleotides in length; (ii) an ASGPR ligand attached at its 3' end, the ASGPR ligand comprising three GalNAc derivatives attached through a trivalent branched linker; (iii) 2'-OMe modifications at positions 1 to 6, 8, and 12 to 21, and 2'-F modifications at positions 7 and 9 to 11; and (iv) phosphorothioate internucleotide linkages between nucleotides 1 and 2 and between nucleotides 2 and 3 (counting from the 5' end) and a sense strand having (b) (i) 23 nucleotides in length; (ii) 2'-OMe modifications at positions 1, 3-5, 7, 8, 10-13, 15, and 17-23, and 2'-F modifications at positions 2, 6, 9, 14, and 16 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotides 1 and 2, between nucleotides 2 and 3, between nucleotides 21 and 22, and between nucleotides 22 and 23 (counting from the 5' end) and an antisense strand having Including, The RNAi agent has a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[0422] In another particular embodiment, the RNAi agent of the invention is (a) (i) 21 nucleotides in length; (ii) an ASGPR ligand attached at its 3' end, the ASGPR ligand comprising three GalNAc derivatives attached through a trivalent branched linker; (iii) 2'-OMe modifications at positions 1 to 6, 8, and 12 to 21, and 2'-F modifications at positions 7 and 9 to 11; and (iv) phosphorothioate internucleotide linkages between nucleotides 1 and 2 and between nucleotides 2 and 3 (counting from the 5' end) and a sense strand having (b) (i) 23 nucleotides in length; (ii) 2'-OMe modifications at positions 1, 3-5, 7, 10-13, 15, and 17-23, and 2'-F modifications at positions 2, 6, 8, 9, 14, and 16 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotides 1 and 2, between nucleotides 2 and 3, between nucleotides 21 and 22, and between nucleotides 22 and 23 (counting from the 5' end) and an antisense strand having Including, The RNAi agent has a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[0423] In another particular embodiment, the RNAi agent of the invention is (a) (i) 19 nucleotides in length; (ii) an ASGPR ligand attached at its 3' end, the ASGPR ligand comprising three GalNAc derivatives attached through a trivalent branched linker; (iii) 2'-OMe modifications at positions 1 to 4, 6, and 10 to 19, and 2'-F modifications at positions 5 and 7 to 9; and (iv) phosphorothioate internucleotide linkages between nucleotides 1 and 2 and between nucleotides 2 and 3 (counting from the 5' end) and a sense strand having (b) (i) 21 nucleotides in length; (ii) 2'-OMe modifications at positions 1, 3-5, 7, 10-13, 15, and 17-21, and 2'-F modifications at positions 2, 6, 8, 9, 14, and 16 (counting from the 5' end); and (iii) phosphorothioate internucleotide linkages between nucleotides 1 and 2, between nucleotides 2 and 3, between nucleotides 19 and 20, and between nucleotides 20 and 21 (counting from the 5' end) and an antisense strand having Including, The RNAi agent has a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[0424] In certain embodiments, the iRNA for use in the methods of the invention is an agent selected from the agents listed in Tables 2, 3, 7, 8, 10, 11, or 13. In one embodiment, the agent is AD-288917. In another embodiment, the agent is AD-288996. In another embodiment, the agent is AD-413639. In one embodiment, the agent is AD-413644. In another embodiment, the agent is AD-413669. These agents may further comprise a ligand. IV. Ligand-Conjugated iRNA

[0425] Another modification of the RNA of the iRNA of the invention involves chemically linking to the RNA one or more ligands, moieties, or conjugates that enhance the activity, cellular distribution, or cellular uptake of the iRNA. Such moieties include, but are not limited to, lipid moieties, such as cholesterol moieties (Letsinger et al., (1989) Proc. Natl. Acid. Sci. USA, 86: 6553-6556), cholic acid (Manoharan et al., (1994) Biorg. Med. Chem. Let., 4: 1053-1060), thioethers, such as beryl-S-tritylthiol (Manoharan et al., (1992) Ann. NY Acad. Sci., 660: 306-309; Manoharan et al., (1993) Biorg. Med. Chem. Let., 3: 2765-2770), thiocholesterol (Oberhauser et al., (1992) Nucl. Acids Res., 20: 533-538), Aliphatic chains, such as dodecanediol or undecyl residues (Saison-Behmoaras et al., (1991) EMBO J, 10: 1111-1118; Kabanov et al., (1990) FEBS Lett., 259: 327-330; Svinarchuk et al., (1993) Biochimie, 75: 49-54), phospholipids, such as di-hexadecyl-rac-glycerol or triethyl-ammonium 1,2-di-O-hexadecyl-rac-glycero-3-phosphonate (Manoharan et al., (1995) Tetrahedron Lett., 36: 3651-3654; Shea et al., (1990) Nucl. Acids Res., 18: 3777-3783), polyamine or polyethylene glycol chains (Manoharan et al., (1995) Nucleosides & Nucleotides, 14: 969-973), or adamantane acetic acid (Manoharan et al., (1995) Tetrahedron Lett., 36: 3651-3654), palmityl moiety (Mishra et al., (1995) Biochim. Biophys. Acta, 1264: 229-237), or octadecylamine or hexylamino-carbonyloxycholesterol (Crooke et al., (1996) J. Pharmacol. Exp. Ther., 277: 923-937 ) etc.

[0426] In one embodiment, the ligand changes the distribution, targeting or life span of the iRNA agent that the ligand is incorporated into.In a preferred embodiment, the ligand provides enhanced affinity to selected targets, for example, molecules, cells or cell types, body compartments, for example, cell compartments or organ compartments, tissues, organs or regions, for example, compared to species that do not have such ligand.Preferred ligands do not participate in duplex pairing in double-stranded nucleic acid.

[0427] Ligands can include naturally occurring substances such as proteins (e.g., human serum albumin (HSA), low-d...

Claims

[Claim 1] The invention as described in the drawings.

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