dsRNA AGENT, OR A SALT THEREOF, PHARMACEUTICAL COMPOSITION, CELL AND IN VITRO METHOD FOR INHIBITING THE EXPRESSION OF 17beta-HYDROXYSTEROID DEHYDROGENASE TYPE 13 (HSD17B13) IN A CELL

Description

dsRNA AGENT, OR A SALT THEREOF, PHARMACEUTICAL COMPOSITION, CELL, AND IN VITRO METHOD FOR INHIBITING THE EXPRESSION OF 17β-HYDROXYSTEROID DEHYDROGENASE TYPE 13 (HSD17B13) IN A CELL Separated from BR112020018758-5, filed on 03 / 20 / 2019 Cross-referencing to related patent applications

[001] This patent application claims the benefit of priority over U.S. Provisional Patent Application No. 62 / 645 941, filed March 21, 2018, U.S. Provisional Patent Application No. 62 / 770 298, filed November 21, 2018, and U.S. Provisional Patent Application No. 62 / 775 590, filed December 5, 2018. The content of each of the foregoing provisional patent applications is incorporated herein in its entirety by reference. Sequence listing

[002] This patent application contains a Sequence Listing that was submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on March 12, 2019, is named 121301-08420_SL.txt and has a size of 1,374,047 bytes. Fundamentals of the invention

[003] 17β-hydroxysteroid dehydrogenase Type 13 (HSD17B13) is part of the 17β-hydroxysteroid dehydrogenase (HSD17B) family of enzymes, whose members have various functions, including, for example, 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 differ in tissue distribution, subcellular location, catalytic preference, and have diverse substrate specificities, as they also catalyze the conversions of substrates other than steroids, such as lipids and retinoids (Marchais-Oberwinkler, et al. (2011)). Petition 870260042207, dated 05 / 05 / 2026, page 18 / 431 / 386 J Steroid Biochem Mol Biol 125(1-2):66-82). HSD17B13 has been shown to enhance hepatic lipogenesis in the liver of normal mice and in cultured human hepatocytes (Su, et al. (2014) Proc Natl Acad Sci USA 111:11437).

[004] Hepatocytes, which form the parenchymal tissue of the liver, are responsible for mobilizing lipids for energy generation and storing excess lipids in the form of lipid droplets (LDs), making the liver the primary organ responsible for lipid homeostasis.

[005] LDs are now recognized as bioactive organelles involved in lipid metabolism, membrane trafficking and signal transduction. Lipid-dependent cells (LDs) are generally composed of a core of neutral lipids (such as triacylglycerols (TGs) and cholesterol esters) surrounded by a phospholipid / cholesterol monolayer. Numerous L-specific proteins associate with the LD membrane and function, for example, to control the flow of molecules into and out of LDs. The predominant proteins associated with hepatocellular LDs belong to the perilipin protein family, but non-perilipin proteins, such as hypoxia-inducible protein 2 (HIG2), patatin-like phospholipase domain-3 (PNPLA3), and HSD17B13, 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).

[006] Increased accumulation of LDs is associated with many metabolic diseases and chronic fibroinflammatory liver diseases, such as hepatic fibrosis, non-alcoholic steatohepatitis (NASH), and non-alcoholic fatty liver disease (NAFLD). HSD17B13 has been identified as one of the most abundantly expressed proteins on the LD surface, specifically located on the surface of LDs in humans and mice with NAFLD. It has also been shown that the expression level of Petition 870260042207, dated 05 / 05 / 2026, p. 19 / 431 / 386 HSD17B13 is upregulated in the liver of patients and mice with NAFLD. Overexpression of HSD17B13 resulted in an increase in the number and size of LDs. Hepatic overexpression of HSD17B13 in C57BL / 6 mice significantly increased lipogenesis and TG content in the livers, leading to a fatty liver phenotype.

[007] There is currently no treatment for chronic fibroinflammatory liver disease. The current standard of care for individuals with chronic fibroinflammatory liver disease includes lifestyle modification and control of associated comorbidities, e.g., hypertension, hyperlipidemia, diabetes, obesity, etc. Consequently, considering that the prevalence of chronic fibroinflammatory liver disease has been progressively increasing in recent years and is projected to increase further, there is a need in the field for alternative treatments for individuals with chronic fibroinflammatory liver disease. Summary of the invention

[008] The present invention provides RNA compositions that effect RNA-induced silencing complex (RISC)-mediated cleavage of RNA transcripts of a 17ε-hydroxysteroid dehydrogenase Type 13 (HSD17B13) gene. The HSD17B13 gene may be located within a cell, e.g., a cell within an individual, such as a human. The present invention also provides methods for using the RNA compositions of the invention to inhibit the expression of an HSD17B13 gene and / or to treat an individual who would benefit from inhibiting or reducing the expression of an HSD17B13 gene, e.g., an individual who suffers from or is prone to suffering from an HSD17B13-associated disease, for example, a chronic fibro-inflammatory liver disease.

[009] Consequently, in one aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent to inhibit the expression of 17ε-hydroxysteroid dehydrogenases type 13 (HSD17B13) in a cell. Petition 870260042207, dated 05 / 05 / 2026, page 20 / 431 / 386 The dsRNA agent includes a sense strand and an antisense strand, wherein the sense strand comprises at least 15 contiguous nucleotides not differing by 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 not differing by more than 1, 2, or 3 nucleotides from the nucleotide sequence of SEQ ID NO:8 or 9. In some embodiments, the dsRNA agent includes a sense strand and an antisense strand, wherein the sense strand comprises at least 15 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:1 or 2, and the antisense strand comprises at least 15 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:8 or 9.

[0010] In another aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent to inhibit the expression of 17β-hydroxysteroid dehydrogenases type 13 (HSD17B13) in a cell. The dsRNA agent includes a sense strand and an antisense strand forming a double-stranded region, wherein said antisense strand comprises a region of complementarity with an mRNA encoding HSD17B13 and comprising at least 15 contiguous nucleotides not differing in more than 1, 2, or 3 nucleotides from any of the antisense sequences listed in any of Tables 2, 3, 7, 8, 10, 11, or 13. In some embodiments, the dsRNA agent includes a sense strand and an antisense strand forming a double-stranded region, wherein said antisense strand comprises a region of complementarity with an mRNA encoding HSD17B13 and comprising at least 15 contiguous nucleotides from any of the antisense sequences listed in any of Tables 2, 3, 7, 8, 10, 11, or 13.

[0011] In one embodiment, the complementarity region comprises at least 15 contiguous nucleotides not differing in more than 1, 2, or 3 nucleotides from any of the nucleotides 213-242; 256-287; 361-385; 447-480; 483-529; 489-529; 630-653; 688-711; 752-777; 753 Petition 870260042207, dated 05 / 05 / 2026, page 21 / 431 / 386 779; 772-806; 781-806; 791-851; 829-858; 870-896; 893-930; 900-930; 910-932; 980-1092; 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 of SEQ ID NO:1. In some embodiments, the complementarity region comprises at least 15 contiguous nucleotides of any one of the 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; 1101-1158; 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 of SEQ ID NO:1.

[0012] In another embodiment, the complementarity region comprises at least 15 contiguous nucleotides not differing in more than 1, 2 or 3 nucleotides from any of the 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, 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; 965987;966-988981-1003; 1005-1027; 1006-1028; 1010-1032; 1005-1032;10521074; 1097-1119;1101-1123; 1102-1124; 1103-1125; 1133-1155; 1135-1157; 1136-1158; 1097-1125;1133-1158; 1176-1198;1188-1210;1243-1265;131513371320-1342; 1322-1344; 1325-1347; 1327-1349;1328-1350; 1320 Petition 870260042207, dated 05 / 05 / 2026, p. 22 / 431 / 386; 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 of SEQ ID NO:1. In some embodiments, the complementarity region comprises at least 15 contiguous nucleotides of any one of the 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, 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; 966-988; 981-1003; 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; 1315-1337; 1320-1342; 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; Petition 870260042207, dated 05 / 05 / 2026, p. 23 / 431 / 386 2188-2210; 2190-2212; 2188-2212; 2194-2216; 2195-2217; 2250-2272; 2232-2254; 2240-2262; 2232-2262; 2242-2264; 2245-2267 2249-2271; 22322271; 2347-2369; 2351-2373; or 2347-2373 of SEQ ID NO:1.

[0013] In another embodiment, the complementarity region comprises at least 15 contiguous nucleotides not differing in more than 1, 2 or 3 nucleotides from any of the 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; 791-817; 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 of SEQ ID NO:1.In some embodiments, the complementarity region comprises at least 15 contiguous nucleotides of any one of the 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; 791-817; 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 of SEQ ID NO:1. In another embodiment, the complementarity region comprises at least 15 contiguous nucleotides not differing in more than 1, 2, or 3 nucleotides from nucleotides 898-930 of SEQ ID NO:1. In some embodiments, the complementarity region comprises at least 15 contiguous nucleotides from nucleotides 898-930 of SEQ ID NO:1.

[0014] In one embodiment, the complementarity region comprises at least 15 contiguous nucleotides not differing in more than 1, 2, or 3 nucleotides from nucleotides 910-932 of SEQ ID NO:1. In some embodiments, the complementarity region comprises by Petition 870260042207, dated 05 / 05 / 2026, page 24 / 431 / 386 minus 15 contiguous nucleotides from nucleotides 910-932 of SEQ ID NO:1.

[0015] In one embodiment, the complementarity region comprises at least 15 contiguous nucleotides not differing in more than 1, 2, or 3 nucleotides from nucleotides 2194-2216 of SEQ ID NO:1. In some embodiments, the complementarity region comprises at least 15 contiguous nucleotides from nucleotides 2194-2216 of SEQ ID NO:1.

[0016] In one embodiment, the complementarity region comprises at least 15 contiguous nucleotides not differing in more than 1, 2, or 3 nucleotides from nucleotides 2242-2264 of SEQ ID NO:1. In some embodiments, the complementarity region comprises at least 15 contiguous nucleotides from nucleotides 2242-2264 of SEQ ID NO:1.

[0017] In one embodiment, the dsRNA agent comprises at least one modified nucleotide.

[0018] In one embodiment, substantially all nucleotides of the sense strand comprise a modification. In another embodiment, substantially all nucleotides of the antisense strand comprise a modification. In yet another embodiment, substantially all nucleotides of the sense strand and substantially all nucleotides of the antisense strand comprise a modification.

[0019] In one aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenases type 13 (HSD17B13) in a cell. The dsRNA agent includes a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides not differing by 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 not differing by more than 1, 2, or 3 nucleotides. Petition 870260042207, dated 05 / 05 / 2026, page 25 / 431 / 386 nucleotides of the nucleotide sequence with SEQ ID NO:8 or 9, wherein substantially all nucleotides of the sense strand and substantially all nucleotides of the antisense strand are modified nucleotides and wherein the sense strand is conjugated to a ligand attached to the 3' end. In some embodiments, the dsRNA agent includes a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:1 or 2 and the antisense strand comprises at least 15 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:8 or 9, wherein substantially all nucleotides of the sense strand and substantially all nucleotides of the antisense strand are modified nucleotides and wherein the sense strand is conjugated to a linker attached to the 3' terminus.

[0020] In one embodiment, all nucleotides of the sense strand comprise a modification. In another embodiment, all nucleotides of the antisense strand comprise a modification. In yet another embodiment, all nucleotides of the sense strand and all nucleotides of the antisense strand comprise a modification.

[0021] In one embodiment, at least one of said modified nucleotides is selected from the group consisting of a deoxynucleotide, a 3'-terminus deoxythymine (dT) nucleotide, a modified 2'-O-methyl nucleotide, a modified 2'-fluorine nucleotide, a modified 2'-deoxy nucleotide, a locked nucleotide, an unlocked nucleotide, a constrained nucleotide, a constrained ethyl nucleotide, an abasic nucleotide, a modified 2'-amino nucleotide, a modified 2'-O-allyl nucleotide, a modified 2'-C-alkyl nucleotide, a modified 2'-hydroxyl nucleotide, a modified 2'-methoxyethyl nucleotide, a modified 2'-O-alkyl nucleotide, a morpholino nucleotide, a phosphoramidate, a nucleotide comprising an unnatural base, a modified nucleotide with Petition 870260042207, dated 05 / 05 / 2026, page 26 / 431 / 386 tetrahydropyran, a nucleotide modified with 1,5-anhydrohexitol, a nucleotide modified with cyclohexenyl, a nucleotide comprising a phosphorothioate group, a nucleotide comprising a methylphosphonate group, a nucleotide comprising a 5'-phosphate, a nucleotide comprising a 5'-phosphate mimetic, a nucleotide modified with glycol, a nucleotide modified with 2-O-(N-methylacetamide) and combinations thereof.

[0022] In one embodiment, the nucleotide modifications are 2'-O-methyl and / or 2'-fluoro modifications.

[0023] The complementarity region can be at least 17 nucleotides long; 19 to 30 nucleotides long; 19-25 nucleotides long; or 21 to 23 nucleotides long.

[0024] Each strand cannot be more than 30 nucleotides long, e.g., each strand is independently 19-30 nucleotides long; each strand is independently 19-25 nucleotides long; each strand is independently 21-23 nucleotides long.

[0025] The dsRNA may include at least one strand comprising a 3' overhang of at least 1 nucleotide; or at least one strand comprising a 3' overhang of at least 2 nucleotides.

[0026] In some embodiments, the dsRNA agent further comprises a linker.

[0027] In one embodiment, the ligand is conjugated to the 3' end of the sense strand of the dsRNA agent.

[0028] In one embodiment, the ligand is a derivative of N-acetylgalactosamine (GalNAc).

[0029] In one modality, the ligand is: Petition 870260042207, dated 05 / 05 / 2026, p. 27 / 431 11 / 386

[0030] In one embodiment, the dsRNA agent is conjugated to the ligand as shown in the following schematic drawing: and, where X is O or S.

[0031] In one modality, X is O.

[0032] In one embodiment, the complementarity region comprises any of the antisense sequences in any of Tables 2, 3, 7, 8, 10, 11, or 13.

[0033] In one aspect, the present invention provides a double strand for inhibiting the expression of 17β-hydroxysteroid dehydrogenases type 13 (HSD17B13) in a cell. The dsRNA agent includes a sense strand complementary to an antisense strand, wherein the antisense strand comprises a region complementary to part of an mRNA encoding HSD17B13, wherein each strand is between approximately 14 and 30 nucleotides in length, wherein said dsRNA agent is represented by formula (III): sense·. 5' np-Na-CXXXji-Nb-YYY-Nb-CZZZjj-Na-nq 3' Petition 870260042207, dated 05 / 05 / 2026, page 28 / 431 / 386 antisense: 3' np'-Na'-(X'X'X')k-Nb'-Y'Y'Y'-Nb'-(Z'Z'Z')lNa'-nq' 5' (III) where: i, j, k, el are each independently 0 or 1; p, p', q, and q' are each independently 0-6; Each Na and Na' independently represents an oligonucleotide sequence comprising 0-25 nucleotides, which are modified or unmodified or combinations thereof, each sequence comprising at least two differently modified nucleotides; Each Nb and Nb' independently represents an oligonucleotide sequence comprising 0-10 nucleotides, which are modified or unmodified or combinations thereof; np, np', nq, and nq', which may or may not each be present, each independently represent a salient nucleotide; XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent a motif of three identical modifications in three consecutive nucleotides; Nb modifications differ from Y modifications, and Nb' modifications differ from Y' modifications; and in which the sense strand is conjugated to at least one ligand.

[0034] 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; kel are both 0; or kel are both 1.

[0035] In one modality, XXX is complementary to X'X'X', YYY is complementary to Y'Y'Y', and ZZZ is complementary to Z'Z'Z'.

[0036] In one embodiment, the YYY motif occurs near or at the cleavage site of the sense strand, e.g., the Y'Y'Y' motif occurs at positions 11, 12, and 13 of the antisense strand from the 5' end. Petition 870260042207, dated 05 / 05 / 2026, p. 29 / 431 / 386

[0037] In one embodiment, formula (III) is represented by formula (IIIa): sense: 5' np-Na-YYY-Na-nq 3' antisense: 3' np'-Na'-Y'Y'Y'-na'-nq' 5' (IIIa).

[0038] In another embodiment, formula (III) is represented by formula (IIIb): sense: 5' np-Na-YYY-Nb-ZZZ-Na-nq 3' antisense: 3' np'-Na'-Y'Y'Y'-Nb'-Z'Z'Z'-na'-nq' 5' (IIIb) wherein each Nb and Nb' independently represents an oligonucleotide sequence comprising 1-5 modified nucleotides.

[0039] In yet another embodiment, formula (III) is represented by formula (IIIc): sense: 5' np-Na-XXX-Nb-YYY-Na-nq 3' antisense: 3' np'-Na'-X'X'X'-Nb'-Y'Y'Y'-na'-nq' 5' (IIIc) wherein each Nb and Nb' independently represents an oligonucleotide sequence comprising 1-5 modified nucleotides.

[0040] In another embodiment, formula (III) is represented by formula (IIId): sense: 5' np-Na-XXX-Nb-YYY-Nb-ZZZ-Na-nq 3' antisense: 3' np'-Na'-X'X'X'- Nb'-Y'Y'Y'-Nb'-Z'Z'Z'-Na'-nq' 5' (IIId) wherein each Nb and Nb' independently represents an oligonucleotide sequence comprising 1-5 modified nucleotides and each Na and Na' independently represents an oligonucleotide sequence comprising 2-10 modified nucleotides.

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

[0042] Each strand cannot have more than 30 nucleotides of Petition 870260042207, dated 05 / 05 / 2026, page 30 / 431 / 386 length, e.g., each strand is independently 19-30 nucleotides long.

[0043] In one embodiment, the nucleotide modifications are 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 nucleotide modifications are 2'-O-methyl or 2'-fluoro modifications.

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

[0046] In one embodiment, at least one strand of the dsRNA agent may comprise 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 linkage between nucleotides.

[0048] In one embodiment, the phosphorothioate or methylphosphonate linkage between nucleotides is at the 3' end of a 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 linkage between nucleotides is at the 5' end of a strand. In one embodiment, the strand is the antisense strand. In another embodiment, the strand is the sense strand.

[0050] In one modality, the tape is the antisense tape. In another modality, the tape is the sense tape.

[0051] In one embodiment, the phosphorothioate or methylphosphonate linkage between nucleotides is at the 5' and 3' ends of a strand.

[0052] In one embodiment, the base pair at position 1 of the 5' end of the duplex antisense tape is an AU base pair.

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

[0054] In one embodiment, q'=0, p=0, q=0 and p' nucleotides Petition 870260042207, dated 05 / 05 / 2026, p. 31 / 431 / 386 salient nucleotides are complementary to the target mRNA. In another embodiment, q'=0, p=0, q=0 and p' salient 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 np' is linked to a neighboring nucleotide by a phosphorothioate bond. In another embodiment, all np' are linked to neighboring nucleotides by phosphorothioate bonds.

[0057] In one embodiment, all nucleotides of the sense strand and all nucleotides of 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 linker is one or more N-acetylgalactosamine (GalNAc) derivatives attached via a monovalent, bivalent, or trivalent branched linker.

[0060] In one modality, the ligand is:

[0061] In one embodiment, the dsRNA agent is conjugated to the ligand as shown in the following schematic drawing: Petition 870260042207, dated 05 / 05 / 2026, p. 32 / 431 16 / 386 where X is O or S.

[0062] In one modality, X is O.

[0063] In one aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenases type 13 (HSD17B13) in a cell. The dsRNA agent includes a sense strand complementary to an antisense strand, wherein the antisense strand comprises a region complementary to a portion of an mRNA encoding HSD17B13, wherein each strand is between approximately 14 and 30 nucleotides in length, wherein the dsRNA agent is represented by 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'-Nb'-(Z'Z'Z')iNa'-Uq' 5' (III) where: i, j, k, and 1 are each independently 0 or 1; p, p', qe q' are each independently 0-6; Each Nae Na' independently represents an oligonucleotide sequence comprising 0-25 nucleotides, which are modified or unmodified or combinations thereof, each sequence comprising at least two differently modified nucleotides; Each Nb and Nb' independently represents a sequence. Petition 870260042207, dated 05 / 05 / 2026, page 33 / 431 / 386 oligonucleotide comprising 0-10 nucleotides, which are modified or unmodified or combinations thereof; Each np, np', nq, and nq', which may or may not be present, independently represents a salient nucleotide; XXX, YYY, ZZZ, X'X'X', Y'Y'Y' and Z'Z'Z' each independently represent a motif of three identical modifications in three consecutive nucleotides, wherein the modifications are either 2'-O-methyl or 2'-fluoro modifications; The modifications on Nb differ from the modification on Y, and the modifications on Nb' differ from the modification on Y'. in which the sense ribbon is conjugated to at least one ligand.

[0064] In one aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenases type 13 (HSD17B13) in a cell. The dsRNA agent includes a sense strand complementary to an antisense strand, wherein the antisense strand comprises a region complementary to a portion of an mRNA encoding HSD17B13, wherein each strand is between approximately 14 and 30 nucleotides in length, wherein the dsRNA agent is represented by formula (III): sense: 5' np-Na-(XXX)i-Nb-YYY-Nb-(ZZZ)j-Na-nq 3' antisense: 3' np'-Na'-(X'X'X')k-Nb'-Y'Y'Y'-Nb'-(Z'Z'Z')lNa'-nq' 5' (III) where: i, j, kel are each independently 0 or 1; Each np, nq, and nq', which may or may not be present, independently represents a salient nucleotide; p, qe q' are each independently 0-6; np' > 0 and at least one np' is linked to a neighboring nucleotide by a phosphorothioate bond; Petition 870260042207, dated 05 / 05 / 2026, page 34 / 431 / 386 each Na and Na' independently represents an oligonucleotide sequence comprising 0-25 nucleotides, which are modified or unmodified or combinations thereof, each sequence comprising at least two differently modified nucleotides; Each Nb and Nb' independently represents an oligonucleotide sequence comprising 0-10 nucleotides, which are modified or unmodified or combinations thereof; XXX, YYY, ZZZ, X'X'X', Y'Y'Y' and Z'Z'Z' each independently represent a motif of three identical modifications in three consecutive nucleotides, wherein the modifications are either 2'-O-methyl or 2'-fluoro modifications; The modifications in Nb differ from the modification in Y, and the modifications in Nb' differ from the modification in Y'. and in which the sense strip is conjugated to at least one ligand.

[0065] In one aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenases type 13 (HSD17B13) in a cell. The dsRNA agent includes a sense strand complementary to an antisense strand, wherein the antisense strand comprises a region complementary to a portion of an mRNA encoding HSD17B13, wherein each strand is between approximately 14 and 30 nucleotides in length, wherein the dsRNA agent is represented by formula (III): sense: 5' np-Na-(XXX)i-Nb-YYY-Nb-(ZZZ)j-Na-nq 3' antisense: 3' np'-Na'-(X'X'X')k-Nb'-Y'Y'Y'-Nb'-(Z'Z'Z')lNa'-nq' 5' (III) where: i, j, kel are each independently 0 or 1; each np, nq and nq', which may or may not be present, Petition 870260042207, dated 05 / 05 / 2026, p. 35 / 431 / 386 independently represents a salient nucleotide; p, qe q' are each independently 0-6; np' > 0 and at least one np' is linked to a neighboring nucleotide by a phosphorothioate bond; Each Na and Na' independently represents an oligonucleotide sequence comprising 0-25 nucleotides, which are modified or unmodified or combinations thereof, each sequence comprising at least two differently modified nucleotides; Each Nb and Nb' independently represents an oligonucleotide sequence comprising 0-10 nucleotides, which are modified or unmodified or combinations thereof; XXX, YYY, ZZZ, X'X'X', Y'Y'Y' and Z'Z'Z' each independently represent a motif of three identical modifications in three consecutive nucleotides, wherein the modifications are either 2'-O-methyl or 2'-fluoro modifications; Nb modifications differ from Y modifications, and Nb' modifications differ from Y' modifications; and wherein the sense strand is conjugated to at least one linker, wherein the linker is one or more GalNAc derivatives attached via a monovalent, bivalent, or trivalent branched linker.

[0066] In one aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenases type 13 (HSD17B13) in a cell. The dsRNA agent includes a sense strand complementary to an antisense strand, wherein the antisense strand comprises a region complementary to a portion of an mRNA encoding HSD17B13, wherein each strand is between approximately 14 and 30 nucleotides in length, wherein the dsRNA agent is represented by formula (III): Petition 870260042207, of 05 / 05 / 2026, p. 36 / 431 / 386 sense: 5' np-Na-(XXX)i-Nb-YYY-Nb-(ZZZ)j-Na-nq 3' antisense: 3' np'-Na'-(X'X'X')k-Nb'-Y'Y'Y'-Nb'-(Z'Z'Z')iNa'-nq' 5' (III) where: i, j, kel are each independently 0 or 1; Each np, nq, and nq', which may or may not be present, independently represents a salient nucleotide; p, q, and q' are each independently 0-6; np' > 0 and at least one np' is linked to a neighboring nucleotide by a phosphorothioate bond; Each Na and Na' independently represents an oligonucleotide sequence comprising 0-25 nucleotides, which are modified or unmodified or combinations thereof, each sequence comprising at least two differently modified nucleotides; Each Nb and Nb' independently represents an oligonucleotide sequence comprising 0-10 nucleotides, which are modified or unmodified or combinations thereof; XXX, YYY, ZZZ, X'X'X', Y'Y'Y' and Z'Z'Z' each independently represent a motif of three identical modifications in three consecutive nucleotides, wherein the modifications are either 2'-O-methyl or 2'-fluoro modifications; The modifications in Nb differ from the modification in Y, and the modifications in Nb' differ from the modification in Y'. wherein the sense strand comprises at least one phosphorothioate linkage; and wherein the sense strand is conjugated to at least one ligand, wherein the ligand is one or more GalNAc derivatives attached via a monovalent, bivalent or trivalent branched linker. Petition 870260042207, dated 05 / 05 / 2026, page 37 / 431 / 386

[0067] In one aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenases type 13 (HSD17B13) in a cell. The dsRNA agent includes a sense strand complementary to an antisense strand, wherein the antisense strand comprises a region complementary to a portion of an mRNA encoding HSD17B13, wherein each strand is between approximately 14 and 30 nucleotides in length, wherein the dsRNA agent is represented by formula (III): sense: 5' np-Na-YYY-na-nq 3' antisense: 3' np'-Na'-Y'Y'Y'-Na'-nq' 5' (IIIa) where: Each np, nq, and nq', which may or may not be present, independently represents a salient nucleotide; p, q, and q' are each independently 0-6; np' > 0 and at least one np' is linked to a neighboring nucleotide by a phosphorothioate bond; Each Na and Na' independently represents an oligonucleotide sequence comprising 0-25 nucleotides, which are modified or unmodified or combinations thereof, each sequence comprising at least two differently modified nucleotides; YYY and Y'Y'Y' each independently represent a motif of three identical modifications in three consecutive nucleotides, wherein the modifications are 2'-O-methyl and / or 2'-fluoro modifications; wherein the sense strand comprises at least one phosphorothioate linkage; and wherein the sense strand is conjugated to at least one ligand, wherein the ligand is one or more GalNAc derivatives attached via a monovalent, bivalent or trivalent branched linker. Petition 870260042207, dated 05 / 05 / 2026, page 38 / 431 / 386

[0068] In one aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenases type 13 (HSD17B13) in a cell. The dsRNA agent includes a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides not differing by 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 not differing by more than 1, 2, or 3 nucleotides from the nucleotide sequence of SEQ ID NO:8 or 9, wherein substantially all nucleotides of 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 linkages between nucleotides at the 5' terminus.wherein substantially all nucleotides of 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 linkages between nucleotides at the 5' end and two phosphorothioate linkages between nucleotides at the 3' end, and wherein the sense strand is conjugated to one or more GalNAc derivatives attached via a monovalent, bivalent, or trivalent branched linker at the 3' end. In some embodiments, the dsRNA agent includes a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:1 or 2 and the antisense strand comprises at least 15 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:8 or 9.wherein substantially all nucleotides of 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 linkages between, Petition 870260042207, dated 05 / 05 / 2026, page 39 / 431 / 386 nucleotides at the 5' end, wherein substantially all nucleotides of 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 linkages between nucleotides at the 5' end and two phosphorothioate linkages between nucleotides at the 3' end, and wherein the sense strand is conjugated to one or more GalNAc derivatives attached via a monovalent, bivalent or trivalent branched linker at the 3' end.

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

[0070] In one embodiment, the complementarity region comprises any of the antisense sequences listed in any 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, AD413639, 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 AD413644. In another embodiment, the agent is AD-413669.

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

[0072] The present invention also provides cells, vectors and pharmaceutical compositions that include any of the dsRNA agents of the invention. The dsRNA agents may be formulated in an unbuffered solution, e.g., saline solution or water, or in a buffered solution, e.g., a solution comprising acetate, citrate, prolamine, carbonate, phosphate or any combination thereof. In one embodiment, the buffered solution is phosphate-buffered saline (PBS). Petition 870260042207, dated 05 / 05 / 2026, page 40 / 431 / 386

[0073] In one aspect, the present invention provides a method for inhibiting the expression of 17ε-hydroxysteroid dehydrogenases type 13 (HSD17B13) in a cell. The method involves bringing the cell into contact with a dsRNA agent or a pharmaceutical composition of the invention, thereby inhibiting the expression of HSD17B13 in the cell.

[0074] The cell can be inside an individual, such as a human.

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

[0076] In one embodiment, the human suffers from a disease, disorder, or condition associated with HSD17B13. In one embodiment, the disease, disorder, or condition associated with HSD17B13 is a chronic fibro-inflammatory liver disease. In one embodiment, the chronic fibro-inflammatory liver disease is selected from the group consisting of liver inflammation, hepatic fibrosis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), liver cirrhosis, alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), HCV-associated 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 an individual. The methods include administering to the individual a therapeutically effective amount of a dsRNA agent or a pharmaceutical composition of the invention, thereby inhibiting the expression of HSD17B13 in the individual.

[0078] In another aspect, the present invention provides a method for treating an individual suffering from a disease, disorder or condition associated with HSD17B13. The method includes administering to the individual a therapeutically effective amount of a dsRNA agent or a composition. Petition 870260042207, dated 05 / 05 / 2026, p. 41 / 431 / 386 pharmaceutical invention, thus treating the individual suffering from a disease, disorder or condition associated with HSD17B13.

[0079] In another aspect, the present invention provides a method for preventing at least one symptom in an individual with a disease, disorder or condition that would benefit from reduced expression of the HSD17B13 gene. The method includes administering to the individual a prophylactically effective amount of a dsRNA agent or a pharmaceutical composition of the invention, thereby preventing at least one symptom in an individual with a disease, disorder or condition that would benefit from reduced expression of the HSD17B13 gene.

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

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

[0082] In one aspect, the present invention provides a method for inhibiting the accumulation of lipid droplets in the liver of an individual suffering from a disease, disorder, or condition associated with HSD17B13. The method includes administering to the individual a therapeutically effective amount of a dsRNA agent or a pharmaceutical composition of the invention and a dsRNA agent targeting a PNPLA3 gene or a pharmaceutical composition comprising an agent targeting a PNPLA3 gene, Petition 870260042207, dated 05 / 05 / 2026, page 42 / 431 / 386, thus inhibiting the accumulation of fat in the liver of an individual suffering from a disease, disorder or condition associated with HSD17B13.

[0083] In another aspect, the present invention provides a method for treating an individual suffering from a disease, disorder or condition associated with HSD17B13. The method includes administering to the individual a therapeutically effective amount of a dsRNA agent or a pharmaceutical composition of the invention and an agent targeting a PNPLA3 gene or a pharmaceutical composition comprising an agent targeting a PNPLA3 gene, thereby treating the individual 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 an individual with a disease, disorder or condition that would benefit from reduced expression of an HSD17B13 gene. The method includes administering to the individual a therapeutically effective amount of a dsRNA agent or a pharmaceutical composition of the invention and an agent targeting a PNPLA3 gene or a pharmaceutical composition comprising an agent targeting a PNPLA3 gene, thereby preventing at least one symptom in an individual with a disease, disorder or condition that would benefit from reduced expression of an HSD17B13 gene.

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

[0086] In another aspect, the present invention provides a method for Petition 870260042207, dated 05 / 05 / 2026, page 43 / 431 / 386 inhibition of the progression of steatosis to steatohepatitis in an individual suffering from steatosis. The method includes administering to the individual a therapeutically effective amount of a dsRNA agent or a pharmaceutical composition of the invention and an agent targeting a PNPLA3 gene or a pharmaceutical composition comprising an agent targeting a PNPLA3 gene, thereby inhibiting the progression of steatosis to steatohepatitis in the individual.

[0087] In one embodiment, administration of the dsRNA agent or pharmaceutical composition to an individual 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 lipid droplet expansion in an individual's liver.

[0088] In one embodiment, the disease, disorder, or condition associated with HSD17B13 is a chronic fibro-inflammatory liver disease.

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

[0090] In one form, chronic fibro-inflammatory liver disease is non-alcoholic steatohepatitis (NASH).

[0091] In one modality, the individual is obese.

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

[0093] In one embodiment, the dsRNA agent is administered to Petition 870260042207, dated 05 / 05 / 2026, page 44 / 431 / 386 individual at a dose between approximately 0.01 mg / kg and 10 mg / kg or between approximately 0.5 mg / kg and 50 mg / kg.

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

[0095] In one embodiment, the methods and uses of the invention further include determining the level of HSD17B13 in the individual.

[0096] In one aspect, the present invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17β-hydroxysteroid dehydrogenases type 13 (HSD17B13) in a cell, wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises a nucleotide sequence of any of the agents in any of Tables 2, 3, 7, 8, 10, 11 or 13, and the antisense strand comprises a nucleotide sequence of any of the agents in any of Tables 2, 3, 7, 8, 10, 11 or 13, wherein substantially all nucleotides of the sense strand and substantially all nucleotides of the antisense strand are modified nucleotides and wherein the dsRNA agent is conjugated to a ligand. Brief description of the drawings

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

[0098] Figure 2A is a graph showing the effect of a dose Petition 870260042207, dated 05 / 05 / 2026, p. 45 / 431 / 386, unique case of AD-288917 in mice expressing human HSD17B13.

[0099] Figure 2B is a graph showing the effect of a single dose of AD-288917 in Cynomolgus monkeys. Detailed description of the invention

[00100] The present invention provides RNA compositions that effect RNA-induced silencing complex (RISC)-mediated cleavage of RNA transcripts of an HSD17B13 gene. The HSD17B13 gene may be located within a cell, e.g., a cell within an individual, such as a human. The present invention also provides methods of using the RNA compositions of the invention to inhibit the expression of an HSD17B13 gene, and to treat an individual who would benefit from inhibiting or reducing the expression of an HSD17B13 gene, e.g., an individual who would benefit from a reduction in liver inflammation., an individual who suffers from or is prone to suffering from a disease, disorder, or condition associated with HSD17B13, such as an individual suffering from or prone to suffering from hepatic fibrosis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), liver cirrhosis, HCV-associated cirrhosis, drug-induced liver injury, and hepatocellular necrosis.

[00101] The iRNAs of the invention targeting HSD17B13 may include an RNA strand (the antisense strand) having a region approximately 30 nucleotides or less in length, e.g., 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, 19-20, 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 Petition 870260042207, dated 05 / 05 / 2026, p. 46 / 431 / 386 nucleotides in length, whose region is substantially complementary to at least part of an mRNA transcript of an HSD17B13 gene.

[00102] In some embodiments, one or both strands of the double-stranded RNAi agents of the invention are up to 66 nucleotides long, for example, 36-66, 26-36, 25-36, 31-60, 22-43, 27-53 nucleotides long, with a region of at least 19 contiguous nucleotides that is substantially complementary to at least a portion of an mRNA transcript of an HSD17B13 gene. In some embodiments, such longer-length antisense stranded RNAi agents include a second RNA strand (the sense strand) 20-60 nucleotides long, wherein the sense and antisense strands form a duplex of 18-30 contiguous nucleotides.

[00103] The use of the iRNA agents described here enables the targeted degradation of mRNAs of an HSD17B13 gene in mammals.

[00104] Very low doses of iRNAs, in particular, can specifically and efficiently mediate RNA interference (RNAi), resulting in significant inhibition of HSD17B13 gene expression. Thus, methods and compositions including these iRNAs are useful for treating an individual who would benefit from inhibiting or reducing HSD17B13 gene expression, e.g., an individual who would benefit from a reduction in liver inflammation, e.g., an individual suffering from or prone to suffering from a disease, disorder, or condition associated with HSD17B13, such as an individual suffering from or prone to suffering from hepatic fibrosis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), liver cirrhosis, HCV-associated cirrhosis, drug-induced liver injury, and hepatocellular necrosis.

[00105] The following detailed description explains how to prepare and use compositions containing iRNAs to inhibit the expression of an HSD17B13 gene, as well as compositions and methods for treating individuals with Petition 870260042207, dated 05 / 05 / 2026, page 47 / 431 / 386 diseases and disorders that would benefit from the inhibition and / or reduction of the expression of this gene. I. Definitions

[00106] In order for the present invention to be more readily understood, certain terms are initially defined. Furthermore, it should be emphasized that whenever a value or range of values ​​of a parameter is mentioned, the intention is that intermediate values ​​and ranges of the mentioned values ​​also form part of this invention.

[00107] The articles “um” and “uma”, in this descriptive report, refer to one or more of (i.e., at least one) of the grammatical object of the article. For example, “um elemento” means one element or more than one element, e.g., a plurality of elements.

[00108] The term “including”, in this descriptive report, means and is used interchangeably with the expression “including, among others”.

[00109] The term “or”, in this descriptive report, means and is used interchangeably with the term “and / or” unless the context clearly indicates otherwise.

[00110] The term “approximately,” in this descriptive report, means within typical tolerance ranges in the technique. For example, “approximately” may be understood as approximately 2 standard deviations from the mean. In certain embodiments, approximately means +10%. In certain embodiments, approximately means +5%. When approximately is present before a series of numbers or a range, it is understood that “approximately” may modify each of the numbers in the series or range.

[00111] The term “HSD17B13”, also known as “hydroxysteroid 17-beta dehydrogenase 13”, “short-chain dehydrogenase / reductase family member 16C”, “short-chain dehydrogenase / reductase 9”, “17-beta-HSD 13”, “17e-HSD13”, “SDR16C3”, Petition 870260042207, dated 05 / 05 / 2026, page 48 / 431 / 386 “SCDR9”, “Member 3, short-chain dehydrogenase / reductase family 16C”, “hydroxysteroid (17-beta) dehydrogenase 13”, “17-beta hydroxysteroid dehydrogenase 13”, “17-beta hydroxysteroid dehydrogenase”, “HMFN0376” and “NIIL497”, refers to the well-known gene encoding a type 13 17β-hydroxysteroid dehydrogenase protein from any vertebrate or mammalian source, including, but not limited to, human, bovine, chicken, rodent, mouse, rat, swine, sheep, primate, monkey and guinea pig, unless otherwise specified.

[00112] The term also refers to fragments and variants of native HSD17B13 that retain at least one in vivo or in vitro activity of a native HSD17B13. The term encompasses complete unprocessed precursor forms of HSD17B13, as well as mature forms resulting from post-translational cleavage of the signal peptide and forms resulting from proteolytic processing.

[00113] Two variants of the human HSD17B13 gene have been previously identified, variant A (or Transcript A) and variant B (or Transcript B). Transcript A includes all seven exons of the HSD17B13 gene, while exon 2 is omitted in Transcript B. The nucleotide and amino acid sequence of a variant A of human HSD17B13 can be found in, for example, GenBank Reference Sequence: NM_178135.4; SEQ ID NO:1); and the nucleotide and amino acid sequence of a variant B of human HSD17B13 can be found in, for example, GenBank Reference Sequence: NM_001136230.2; SEQ ID NO:2. As described in US Patent ApplicationIn Patent Application No. 15 / 875,514, filed January 19, 2018, and PCT Patent Application No. PCT / US2018 / 014357, filed January 19, 2018 (the contents of which are incorporated herein in their entirety by reference), six additional transcripts of HSD17B13 that are expressed (CH, SEQ ID NOS: 17, 18, 19, 20, 21 and 22, respectively) have been identified. In Transcript C, exon 6 is omitted. Petition 870260042207, dated 05 / 05 / 2026, page. 49 / 431 / 386 when compared to Transcript A. In Transcript D, there is an insertion of a 3' guanine from exon 6, resulting in a frameshift (read-through) and premature truncation of exon 7 when compared to Transcript A. In Transcript E, there is an additional exon between exons 3 and 4 when compared to Transcript A. In Transcript F, which is expressed only in carriers of the HSD17B13 rs72613567 variant, there is a read-through transreading of exon 6 to intron 6 when compared to Transcript A. In Transcript G, exon 2 is omitted, and there is an insertion of a 3' guanine from exon 6, resulting in a frameshift and premature truncation of exon 7 when compared to Transcript A. In Transcript H, there is an additional exon between exons 3 and 4, and there is an insertion of a 3' guanine from exon 6, resulting in a frameshift and premature truncation of exon 7 when compared to Transcript A.

[00114] An additional HSD17B13 transcript that is expressed at low levels (F', SEQ ID NO: 23) has also been identified. Similar to Transcript F, Transcript F' also includes an exon 6 to intron 6 transreading when compared to Transcript A, but unlike Transcript F, the transreading does not include the inserted thymine present in the variant HSD17B13 gene rs72613567. The nucleotide positions of the exons within the HSD17B13 genes for each Transcript are provided below.

[00115] SEQ ID NO:15 is the nucleotide sequence of the Wild Type HSD17B13 Genomic Sequence (Human Genome Assembly GRCh38) and SEQ ID NO: 16 is the nucleotide sequence of the Variant HSD17B13 Genomic Sequence (Human Genome Assembly GRCh38; rs72613567- T insertion in chr4: 87310241-87310240): T insertion at position 12666.

[00116] Nucleotide positions in SEQ ID NO: 15 for exons of the most prevalent transcripts of HSD17B13 in individuals homozygous for the wild-type HSD17B13 gene Transcribed A Transcribed B Transcribed E Transcribed F' Exon 1 1-275 1-275 1-275 1-275 Petition 870260042207, dated 05 / 05 / 2026, p. 50 / 431 / 386 Exon 2 4471-4578 omitted 4471-4578 4471-4578 Exon 3 5684-5815 5684-5815 5684-5815 5684-5815 Exon 3' not present not present 6210-6281 not present Exon 4 7308-7414 7308-7414 7308-7414 7308-7414 Exon 5 8947-9084 8947-9084 8947-9084 8947-9084 Exon 6 12548-12664 12548-12664 12548-12664 12548-13501* Exon 7 17599-19118 17599-19118 17599-19118 omitted *Includes transreading from exon 6 to intron 6; transreading = positions 12665-13501

[00117] Nucleotide positions in SEQ ID NO: 16 for exons of the most prevalent transcripts of HSD17B13 in individuals homozygous for the variant gene rs72613567 HSD17B13 (T insertion at position 12666). Transcribed C Transcribed D Transcribed F Transcribed G Transcribed H Exon 1 1-275 1-275 1-275 1-275 1-275 Exon 2 4471-4578 4471-4578 4471-4578 omitted 4471-4578 Exon 3 5684-5815 5684-5815 5684-5815 5684-5815 5684-5815 Exon 3' not present not present not present not present 6210-6281 Exon 4 7308-7414 7308-7414 7308-7414 7308-7414 7308-7414 Exon 5 8947-9084 8947-9084 8947-9084 8947-9084 8947-9084 Exon 6 omitted 12548-12665Λ 12548-13502* 12548-12665Λ 12548-12665Λ Exon 7 17600-19119 17600-19119 omitted 17600-19119 17600-19119 AIncludes the additional residue 12665 ​​at the 3' end when compared to Transcript A *Includes transreading from exon 6 to intron 6; transreading = positions 12665-13502

[00118] There are two variants of the mouse HSD17B13 gene; the nucleotide and amino acid sequence of a mouse Hsd17b13, transcript 1 variant, can be found in, for example, GenBank Reference Sequence: NM_001163486.1; SEQ ID NO:3); and the nucleotide and amino acid sequence of a mouse Hsd17b13, transcript 2 variant, can be found in, for example, GenBank Reference Sequence: NM_198030.2; SEQ ID NO:4. The nucleotide and amino acid sequence of a rat Hsd17b13 gene can be found in, for example, GenBank Reference Sequence: NM_001009684.1; SEQ ID NO:5). The nucleotide and amino acid sequence of an HSD17B13 gene from Macaca mulatta can be found in, for example, GenBank Reference Sequence: XM_015138766.1; SEQ ID NO:6). The nucleotide and amino acid sequence Petition 870260042207, dated 05 / 05 / 2026, p. 51 / 431 / 386 amino acids of an HSD17B13 gene from Macaca fascicularis can be found in, for example, GenBank Reference Sequence: XM_005555367.2; SEQ ID NO:7).

[00119] Additional examples of HSD17B13 mRNA sequences are readily available in publicly available databases, e.g. (for example, GenBank, UniProt and OMIM).

[00120] The term “HSD17B13”, in this descriptive report, also refers to a particular polypeptide expressed in a cell by naturally occurring variations in the DNA sequence of the HSD17B13 gene, such as a single nucleotide polymorphism (SNP) in the HSD17B13 gene. Numerous SNPs within the HSD17B13 gene have been identified and can be found at, for example, NCBI dbSNP (see, for example, www.ncbi.nlm.nih.gov / snp).

[00121] In this descriptive report, “target sequence” refers to a contiguous portion of the nucleotide sequence of an mRNA molecule formed during the transcription of an HSD17B13 gene, including mRNA that is produced by RNA processing of a primary transcription product. In one embodiment, the target portion of the sequence will be at least long enough to serve as a substrate for iRNA-directed cleavage near or at that portion of the nucleotide sequence of an mRNA molecule formed during the transcription of an HSD17B13 gene.

[00122] The target sequence of an HSD17B13 gene can be approximately 9-36 nucleotides long, for example. (e.g., approximately 15-30 nucleotides long. For example, the target sequence can be approximately 15-30 nucleotides, 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 Petition 870260042207, dated 05 / 05 / 2026, page 52 / 431 / 386 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23 or 21-22 nucleotide lengths. Intermediate intervals and lengths to the intervals and lengths mentioned above are also contemplated as part of the invention.

[00123] In this descriptive report, the term “strand comprising a sequence” refers to an oligonucleotide comprising a chain of nucleotides that is described by the referred sequence using standard nucleotide nomenclature.

[00124] “G”, “C”, “A”, “T”, and “U” generally represent a nucleotide containing guanine, cytosine, adenine, thymidine, and uracil as its base, respectively. However, it will be understood that the term “ribonucleotide” or “nucleotide” may also refer to a modified nucleotide, as described in more detail below, or an alternative substitute moiety (see, e.g., Table 1). Those skilled in the art are well aware that guanine, cytosine, adenine, and uracil can be substituted for other moieties without substantially altering the base-pairing properties of an oligonucleotide comprising a nucleotide bearing such a substitution moiety. For example, without limitation, a nucleotide comprising inosine as its base can base-pair with nucleotides containing adenine, cytosine, or uracil.Consequently, nucleotides containing uracil, guanine, or adenine can be substituted in the nucleotide sequences of the dsRNA presented in the invention by a nucleotide containing, for example, inosine. In another example, adenine and cytosine anywhere in the oligonucleotide can be substituted by guanine and uracil, respectively, to form a GU wobble base pairing with the target mRNA. Sequences containing such substitution moieties are suitable for the compositions and methods presented in the invention.

[00125] The terms “iRNA”, “RNAi agent”, “iRNA agent”, “agent Petition 870260042207, dated 05 / 05 / 2026, p. 53 / 431 / 386 RNA interference, as used interchangeably in this descriptive report, refers to an agent containing RNA as defined herein, and acting as a mediator in the targeted cleavage of an RNA transcript via the RNA-induced silencing complex (RISC) pathway. The iRNA directs the specific degradation of the mRNA sequence through a process known as RNA interference (RNAi). The iRNA modulates, e.g., inhibits, the expression of the HSD17B13 gene in a cell, e.g., a cell within an individual, such as a mammal.

[00126] In one embodiment, an RNAi agent of the invention includes a single-stranded RNA that interacts with a target RNA sequence, e.g., an HSD17B13 target mRNA sequence, to direct the cleavage of the target RNA. Without intending to be theory-bound, it is believed that the long double-stranded RNA introduced into cells is broken down into siRNA by a type of Type III endonuclease known as Dicer (Sharp et al. (2001) Genes Dev. 15:485). Dicer, a ribonuclease-III type enzyme, processes dsRNA into short interfering RNAs of 19-23 base pairs with two characteristic 3' base overhangs (Bernstein, et al., (2001) Nature 409: 363). The siRNAs are then incorporated into an RNA-induced silencing complex (RISC), in which one or more helicases unwind the siRNA complex, enabling the complementary antisense strand to guide target recognition (Nykanen, et al., (2001) Cell 107:309).Upon binding to the appropriate target mRNA, one or more endonucleases within the RISC cleave the target to induce silencing (Elbashir, et al., (2001) Genes Dev. 15:188). Thus, in one aspect, the invention relates to a single-stranded RNA (sssiRNA) generated within a cell that promotes the formation of a RISC complex to effect the silencing of the target gene, namely, an HSD17B13 gene. Consequently, the term “siRNA” is also used herein to refer to an RNAi as described above.

[00127] In another embodiment, the RNAi agent may be an agent Petition 870260042207, dated 05 / 05 / 2026, p. 54 / 431 / 386 Single-stranded RNAi is introduced into a cell or organism to inhibit a target mRNA. Single-stranded RNAi agents (ssRNAi) bind to the RISC endonuclease, Argonaute 2, which then cleaves the target mRNA. Single-stranded siRNAs typically have 15-30 nucleotides and are chemically modified. The design and testing of single-stranded RNAi agents are described in U.S. Patent No. 8,101,348 and in Lima et al., (2012) Cell 150: 883-894, the contents of which are incorporated herein in their entirety by reference. Any of the antisense nucleotide sequences described herein may be used as a single-stranded siRNA as described herein or as chemically modified by the methods described in Lima et al., (2012) Cell 150: 883-894.

[00128] In another embodiment, an “iRNA” for use in the compositions and methods of the invention is a double-stranded RNA and is referred to herein as a “double-stranded RNAi agent”, “double-stranded RNA (dsRNA) molecule”, “dsRNA agent” or “dsRNA”. The term “dsRNA” refers to a complex of ribonucleic acid molecules having a duplex structure comprising two antiparallel and substantially complementary strands of nucleic acid, referred to as having “sense” and “antisense” orientations with respect to a target RNA, i.e., an HSD17B13 gene. In some embodiments of the invention, a double-stranded RNA (dsRNA) triggers the degradation of a target RNA, e.g., an mRNA, by means of a posttranscriptional gene silencing mechanism, referred to herein as RNA interference or RNAi.

[00129] In general, most nucleotides in each strand of a dsRNA molecule are ribonucleotides, but, as described in detail below, each or both strands may also include one or more non-ribonucleotides, e.g., a deoxyribonucleotide and / or a modified nucleotide. Furthermore, in this descriptive report, an “RNAi agent” may include ribonucleotides with chemical modifications; an RNAi agent may include substantial modifications in multiple nucleotides. Petition 870260042207, dated 05 / 05 / 2026, page 55 / 431 / 386 In this descriptive report, the term “modified nucleotide” refers to a nucleotide having, independently, a modified sugar moiety, a modified nucleotide linkage, and / or a modified nucleobase. Thus, the term modified nucleotide encompasses substitutions, additions, or removal of, e.g., a functional group or atom, to nucleoside linkages, sugar moieties, or nucleobases. Modifications suitable for use in the agents of the invention include all types of modifications described herein or known in the art. Any such modification, as used in an siRNA-type molecule, is encompassed by “RNAi agent” for purposes of this descriptive report and the claims.

[00130] The duplex region can be of any length that allows for the specific degradation of a desired target RNA via a RISC pathway, and can range from approximately 9 to 36 base pairs in length, p. e.g., approximately 15-30 base pairs of length, for example, approximately 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 of length, such as approximately 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, 19-20, 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 length pairs. Intermediate intervals and lengths to the intervals and lengths mentioned above are also contemplated as part of the invention.

[00131] The two strands that form the duplex structure can be different portions of a larger RNA molecule, or they can be separate RNA molecules. When the two strands are part of a larger molecule and are therefore connected by an unbroken chain of nucleotides between the 3' end of one strand and the 5' end of the other strand. Petition 870260042207, dated 05 / 05 / 2026, page 56 / 431 / 386, which forms the duplex structure, the RNA connecting strand is referred to as a “hairpin loop”. A hairpin loop may comprise at least one unpaired nucleotide. In some embodiments, the hairpin loop may comprise 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.

[00132] When the two substantially complementary strands of a dsRNA are composed of separate RNA molecules, these molecules do not need to, but can be, covalently connected. When the two strands are covalently connected by means other than an unbroken chain of nucleotides between the 3' end of one strand and the 5' end of the respective other strand that forms the duplex structure, the connecting structure is referred to as a “linker”. The RNA strands may have the same or a different number of nucleotides. The maximum number of base pairs is the number of nucleotides in the shorter strand of the dsRNA minus any overhangs present in the duplex. In addition to the duplex structure, an RNAi may comprise one or more nucleotide overhangs.

[00133] In one embodiment, an RNAi agent of the invention is a dsRNA whose strands comprise fewer than 30 nucleotides, e.g., 17-27, 19-27, 17-25, 19-25, or 19-23, which interacts with a target RNA sequence, e.g., an HSD17B13 target mRNA sequence, to direct the cleavage of the target RNA. In another embodiment, an RNAi agent of the invention is a dsRNA whose strands comprise 19-23 nucleotides, which interacts with a target RNA sequence, e.g., an HSD17B13 target mRNA sequence, to direct the cleavage of the target RNA. In one embodiment, the sense strand is 21 nucleotides long. In another embodiment, the antisense strand is 23 nucleotides long.

[00134] In this descriptive report, the term “nucleotide salience” Petition 870260042207, dated 05 / 05 / 2026, page 57 / 431 / 386 refers to at least one unpaired nucleotide that protrudes from the duplex structure of an iRNA, e.g., a dsRNA. For example, when a 3' end of one strand of a dsRNA extends beyond the 5' end of the other strand, or vice versa, there is a nucleotide overhang. A dsRNA may comprise an overhang of at least one nucleotide; alternatively, the overhang may comprise at least two nucleotides, at least three nucleotides, at least four nucleotides, at least five nucleotides, or more. A nucleotide overhang may comprise or consist of a nucleotide / nucleoside analog, including a deoxynucleotide / nucleoside. The protrusion(s) may be on the sense tape, the antisense tape, or any combination thereof.Furthermore, the nucleotide(s) of a protrusion may be present at the 5' end, the 3' end, or both ends of an antisense or sense strand of a dsRNA.

[00135] In one embodiment, the antisense strand of a dsRNA has a 1-10 nucleotide overhang, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides, at the 3' and / or 5' end. In another embodiment, one or more of the nucleotides in the overhang are replaced by a thiophosphate nucleoside.

[00136] In certain embodiments, the overhang on the sense strand or the antisense strand, or both, may include extended lengths longer than 10 nucleotides, e.g., 10⁻³⁰ nucleotides, 10⁻²⁵ nucleotides, 10⁻²⁰ nucleotides, or 10⁻¹⁵ nucleotides in length. In certain embodiments, there is an extended overhang on the sense strand of the duplex. In certain embodiments, an extended overhang is present at the 3' end of the sense strand of the duplex. In certain embodiments, an extended overhang is present at the 5' end of the sense strand of the duplex. In certain embodiments, there is a Petition 870260042207, dated 05 / 05 / 2026, page 58 / 431 / 386 extended overhang on the antisense strand of the duplex. In certain embodiments, an extended overhang is present at the 3' end of the antisense strand of the duplex. In certain embodiments, an extended overhang is present at the 5' end of the antisense strand of the duplex. In certain embodiments, one or more of the nucleotides in the extended overhang are replaced by a thiophosphate nucleoside.

[00137] The terms “blunt” or “blunt-ended,” used in this descriptive report in reference to a dsRNA, means that there are no unpaired nucleotides or nucleotide analogs at a given terminal end of a dsRNA, i.e., no nucleotide overhang. One or both ends of a dsRNA may be blunt. When both ends of a dsRNA are blunt, the dsRNA is said to have blunt ends. To clarify, a dsRNA with “blunt ends” is a dsRNA that is blunt at both ends, i.e., it has no nucleotide overhang at either end of the molecule. Most often, such a molecule will be double-stranded along its entire length.

[00138] The term “antisense strand” or “guide strand” refers to the strand of an iRNA, e.g. (for example, a dsRNA), that includes a region that is substantially complementary to a target sequence, e.g., an HSD17B13 mRNA.

[00139] In this descriptive report, the term “complementarity region” refers to the region in the antisense strand that is substantially complementary to a sequence, for example, a target sequence, e.g., a nucleotide sequence of HSD17B13, as defined herein. When the complementarity region is not fully complementary to the target sequence, mismatches may occur in the internal or terminal regions of the molecule. In general, the most tolerated mismatches are in the terminal regions, e.g., within 5, 4, 3, or 2 nucleotides of the 5' and / or 3' end of the iRNA.

[00140] The term “sense tape” or “passing tape” in this report Petition 870260042207, dated 05 / 05 / 2026, page 59 / 431 / 386, descriptive, refers to the strand of an iRNA that includes a region that is substantially complementary to a region of the antisense strand as that term is defined herein.

[00141] In this descriptive report, the term “cleavage region” refers to a region that is located immediately adjacent to the cleavage site. The cleavage site is the site on the target where cleavage occurs. In some embodiments, the cleavage region comprises three bases at each end and immediately adjacent to the cleavage site. In some embodiments, the cleavage region comprises two bases at each end and immediately adjacent to the cleavage site. In some embodiments, the cleavage site occurs specifically at the site bound by nucleotides 10 and 11 of the antisense strand, and the cleavage region comprises nucleotides 11, 12, and 13.

[00142] In this descriptive report, and unless otherwise indicated, the term “complementary,” when used to describe a first nucleotide sequence in relation to a second nucleotide sequence, refers to the ability of an oligonucleotide or polynucleotide comprising the first nucleotide sequence to hybridize and form a duplex structure under certain conditions with an oligonucleotide or polynucleotide comprising the second nucleotide sequence, as will be understood by those skilled in the art. Such conditions may be, for example, stringent conditions, where stringent conditions may include: 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mM EDTA, 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 physiologically relevant conditions, as may be found inside an organism, may apply.The expert in the field will be able to determine the most appropriate set of conditions for a complementarity test of two. Petition 870260042207, dated 05 / 05 / 2026, p. 60 / 431 / 386 sequences according to the final application of the hybridized nucleotides.

[00143] Complementary sequences within an iRNA, e.g., within a dsRNA as described herein, include the base pairing of the oligonucleotide or polynucleotide comprising a first nucleotide sequence with an oligonucleotide or polynucleotide comprising a second nucleotide sequence along the entire length of one or both nucleotide sequences. Such sequences may be referred to as “fully complementary” with respect to each other. However, when a first sequence is referred to as “substantially complementary” with respect 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 base pairing errors when hybridizing to a duplex of up to 30 base pairs, while retaining the ability to hybridize under the conditions most relevant to their final application, e.g.Inhibition of gene expression via a RISC pathway. However, when two oligonucleotides are designed to form, upon hybridization, one or more single-stranded overhangs, such overhangs will not be considered pairing errors with respect to the determination of complementarity. For example, a dsRNA comprising one oligonucleotide 21 nucleotides long and another oligonucleotide 23 nucleotides long, wherein the longer oligonucleotide comprises a 21-nucleotide sequence that is fully complementary to the shorter oligonucleotide, may still be referred to as “fully complementary” for the purposes described herein.

[00144] “Complementary” sequences, in this descriptive report, may also include, or be formed entirely of, non-Watson-Crick base pairs and / or base pairs formed of non-natural and modified nucleotides, provided that the above requirements regarding their capability are met. Petition 870260042207, dated 05 / 05 / 2026, page 61 / 431 / 386, to ensure hybridization is fulfilled. Such non-Watson-Crick base pairs include, among others, G:U Wobble or Hoogstein base pairings.

[00145] The terms “complementary”, “fully complementary” and “substantially complementary”, in this descriptive report, may be used with respect to the correct base matching between the sense strand and the antisense strand of a dsRNA, or between the antisense strand of an iRNA agent and a target sequence, as will be understood from the context of their use.

[00146] In this descriptive report, a polynucleotide that is “substantially complementary to at least part of” a messenger RNA (mRNA) refers to a polynucleotide that is substantially complementary to a contiguous portion of the mRNA of interest (e.g., an mRNA encoding HSD17B13). For example, a polynucleotide is complementary to at least part of an HSD17B13 mRNA if the sequence is substantially complementary to an unbroken portion of an mRNA encoding HSD17B13.

[00147] Consequently, in some embodiments, the antisense strand polynucleotides described herein are fully complementary to the target sequence of HSD17B13. In other embodiments, the antisense strand polynucleotides described herein are substantially complementary to the target sequence of HSD17B13 and comprise a contiguous nucleotide sequence that is at least about 80% complementary along its entire length to the equivalent region of the nucleotide sequence of SEQ ID NO:1, or a fragment of SEQ ID NO:1, such as approximately 85%, approximately 86%, approximately 87%, approximately 88%, approximately 89%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% complementary. Petition 870260042207, dated 05 / 05 / 2026, page 62 / 431 / 386

[00148] In one embodiment, an RNAi agent of the invention includes a sense strand that is substantially complementary to an antisense polynucleotide that, in turn, is complementary to a target sequence of HSD17B13, and wherein the sense strand polynucleotide comprises a contiguous nucleotide sequence that is at least about 80% complementary along its entire length to the equivalent region of the nucleotide sequence of SEQ ID NO:8, or any fragment of SEQ ID NO:8, such as approximately 85%, approximately 86%, approximately 87%, approximately 88%, approximately 89%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98% or approximately 99% complementary.

[00149] In some embodiments, an iRNA of the invention includes an antisense strand that is substantially complementary to the target sequence of HSD17B13 and comprises a contiguous nucleotide sequence that is at least about 80% complementary along its entire length to the equivalent nucleotide sequence region of any of the sense strands in any of Tables 2, 3, 7, 8, 10, 11 or 13, or a fragment of any of the sense strands in any of Tables 2, 3, 7, 8, 10, 11 or 13, such as approximately 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% complementary or 100% complementary.

[00150] The term “inhibit”, in this descriptive report, is used interchangeably with “reduce”, “silence”, “negatively regulate”, “suppress” and other similar terms, and includes any level of inhibition.

[00151] The term “inhibition of the expression of an HSD17B13 gene” in this descriptive report includes the inhibition of the expression of any HSD17B13 gene (such as, for example, the mouse HSD17B13 gene, gene Petition 870260042207, dated 05 / 05 / 2026, p. 63 / 431 / 386 Mouse HSD17B13, monkey HSD17B13 gene, or human HSD17B13 gene) as well as variants or mutants of an HSD17B13 gene that encodes an HSD17B13 protein.

[00152] “Inhibition of HSD17B13 gene expression” includes any level of inhibition of an HSD17B13 gene, e.g., at least partial suppression of HSD17B13 gene expression, such as inhibition by at least approximately 20%. In certain embodiments, the inhibition is at least approximately 25%, at least approximately 30%, at least at least approximately 35%, at least approximately 40%, at least approximately 45%, at least approximately 50%, at least approximately 55%, at least approximately 60%, at least approximately 65%, at least approximately 70%, at least approximately 75%, at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 91%, at least approximately 92%, at least approximately 93%, at least approximately 94%, at least approximately 95%, at least approximately 96%, by less approximately 97%, at least approximately 98%, or at least approximately 99%.

[00153] The expression of an HSD17B13 gene can be assessed based on the level of any variable associated with HSD17B13 gene expression, e.g., HSD17B13 mRNA level or HSD17B13 protein level. HSD17B13 gene expression can also be assessed indirectly based on, for example, circulating alanine aminotransferase (ALT) levels, or HSD17B13 enzyme activity in a tissue sample, such as a liver sample. Inhibition can be assessed by a decrease in an absolute or relative level of one or more of these variables compared to a control level. The control level can be any type of control level used in the technique, e.g., Petition 870260042207, dated 05 / 05 / 2026, p. 64 / 431 / 386 a pre-dose baseline level or a determined level of an individual, cell or similar sample that is not treated or treated with a control (such as, e.g., buffer-only control or inactive agent control).

[00154] In one embodiment, at least partial suppression of the expression of an HSD17B13 gene is assessed by a reduction in the amount of HSD17B13 mRNA, which can be isolated or detected in a first cell or group of cells in which an HSD17B13 gene is transcribed and which has been treated in such a way that the expression of an HSD17B13 gene is inhibited, compared to a second cell or group of cells substantially identical to the first cell or group of cells, but which has not been treated in this way (control cells).

[00155] The degree of inhibition can be expressed in terms of: (mRNA in control cells) - (mRNA in treated cells) · 100% (mRNA in control cells)

[00156] The expression “bringing a cell into contact with an RNAi agent”, such as a dsRNA, in this descriptive report, includes bringing a cell into contact by any possible means. Contacting a cell with an RNAi agent includes contacting a cell in vitro with the iRNA or contacting a cell in vivo with the iRNA. Contact can be made directly or indirectly. Thus, for example, the RNAi agent can be brought into physical contact with the cell by the individual performing the method or, alternatively, the RNAi agent can be placed in a situation that will allow or cause it to subsequently come into contact with the cell.

[00157] In vitro cell contact can be achieved, for example, by incubating the cell with the RNAi agent. In vivo cell contact can be achieved, for example, by injecting the RNAi agent into or near the tissue where the cell is located, or by injecting the RNAi agent into another area, e.g., the bloodstream or subcutaneous space, such that the agent Petition 870260042207, dated 05 / 05 / 2026, page 65 / 431 / 386, subsequently reaches the tissue where the cell to be contacted is located. For example, the RNAi agent may contain and / or be attached to a ligand, e.g., GalNAc3, which directs the RNAi agent to a site of interest, e.g., the liver. Combinations of in vitro and in vivo methods for contact are also possible. For example, a cell may also be placed in vitro in contact with an RNAi agent and subsequently transplanted into an individual.

[00158] In one embodiment, cell contact with an iRNA includes “introducing” or “delivering the iRNA into the cell” by facilitating or effecting uptake or absorption into the cell. Uptake or absorption of an iRNA can occur through unaided or active diffusive cellular processes, or by auxiliary agents or devices. The introduction of an iRNA into a cell can be in vitro and / or in vivo. For example, for in vivo introduction, the iRNA can be injected into a tissue site or administered systemically. In vivo delivery can also be accomplished by a beta-glucan delivery system, such as those described in U.S. Patents Nos. 5,032,401 and 5,607,677, and U.S. Publication No. 2005 / 0281781, the contents of which are incorporated herein in their entirety by reference. In vitro introduction into a cell includes methods known in the art such as electroporation and lipofection. Additional approaches are described below and / or are known in the art.

[00159] The term “lipid nanoparticle” or “LNP” refers to a vesicle comprising a lipid layer encapsulating a pharmaceutically active molecule, such as a nucleic acid molecule, e.g., an iRNA or plasmid from which an iRNA is transcribed. LNPs are described in, for example, U.S. Patents Nos. 6,858,225, 6,815,432, 8,158,601 and 8,058,069, the contents of which are incorporated herein in their entirety by reference.

[00160] In this descriptive report, an “individual” is an animal, such Petition 870260042207, dated 05 / 05 / 2026, p. 66 / 431 / 386 as a mammal, including a primate (such as a human, a non-human primate, e.g., a monkey and a chimpanzee), a non-primate (such as 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).

[00161] In one embodiment, the individual is a human, such as a human being treated or evaluated for a disease, disorder, or condition that would benefit from reduced HSD17B13 expression; a human at risk of a disease, disorder, or condition that would benefit from reduced HSD17B13 expression; a human having a disease, disorder, or condition that would benefit from reduced HSD17B13 expression; and / or a human being treated for a disease, disorder, or condition that would benefit from reduced HSD17B13 expression, as described herein.

[00162] In one embodiment, the individual is heterozygous for the gene encoding the I148M variant of the patatin-like phospholipase domain-containing protein 3 (PNPLA3). In another embodiment, the individual is homozygous for the gene encoding the I148M variant of PNPLA3. In one embodiment, the individual is heterozygous for the gene encoding the I144M variant of the patatin-like phospholipase domain-containing protein 3 (PNPLA3). In another embodiment, the individual is homozygous for the gene encoding the I144M variant of PNPLA3. In one embodiment, the individual is homozygous for the gene encoding a functional HSD17B13 protein. In another embodiment, the individual is heterozygous for the gene encoding a functional HSD17B13 protein. In yet another variant, the individual is heterozygous for the gene that codes for a functional HSD17B13 protein and a gene that codes for a loss-of-function variant of HSD17B13.In another form, the individual carries the variant HSD17B13 rs72613567, e.g., HSD17B13 rs72613567:TA. Petition 870260042207, dated 05 / 05 / 2026, p. 67 / 431 / 386

[00163] In this descriptive report, the terms “treat” or “treatment” refer to a beneficial or desired outcome including, but not limited to, relief or improvement of one or more symptoms associated with HSD17B13 gene expression and / or HSD17B13 protein production, e.g., an HSD17B13-associated disease, such as a chronic fibro-inflammatory liver disease, e.g., liver inflammation, hepatic fibrosis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), liver cirrhosis, alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), HCV-associated cirrhosis, drug-induced liver injury, hepatocellular necrosis, and / or hepatocellular carcinoma. “Treatment” may also mean prolonging survival compared to predicted survival in the absence of treatment.

[00164] The term “lower” in the context of a disease associated with HSD17B13 refers to a statistically significant decrease in that level. The decrease may 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 modalities, a decrease is at least 20%. “Lowering” in the context of the HSD17B13 level in an individual is preferably to descend to a level accepted as within the normal range for an individual without such a disorder.

[00165] In this descriptive report, “prevention” or “prevent,” when used in reference to a disease, disorder, or condition that would benefit from a reduction in the expression of an HSD17B13 gene, refers to a reduction in the likelihood that an individual will develop a symptom associated with such disease, disorder, or condition, e.g., a symptom of HSD17B13 gene expression, such as liver inflammation, fibrosis. Petition 870260042207, dated 05 / 05 / 2026, page 68 / 431 / 386 hepatic, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), liver cirrhosis, alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), HCV-associated cirrhosis, drug-induced liver injury, hepatocellular necrosis and / or hepatocellular carcinoma. The non-development of a disease, disorder or condition, or the reduction in the development of a symptom associated with such disease, disorder or condition (e.g., by at least approximately 10% on a clinically accepted scale for that disease or disorder), or the delayed display (e.g., by days, weeks, months or years) of late symptoms (e.g., reduction in lipid accumulation in the liver and / or expansion of lipid droplets in the liver) is considered effective prevention.

[00166] In this descriptive report, the term “HSD17B13-associated disease” refers to a disease or disorder that is caused by or associated with HSD17B13 gene expression or HSD17B13 protein production. The term “HSD17B13-associated disease” includes a disease, disorder, or condition that would benefit from a decrease in HSD17B13 gene expression or protein activity.

[00167] In one embodiment, an “HSD17B13-associated disease” is a chronic fibro-inflammatory liver disease. A “chronic fibro-inflammatory liver disease” is any disease, disorder, or condition associated with chronic liver inflammation and / or fibrosis. Non-limiting examples of chronic fibro-inflammatory liver disease include, for example, liver inflammation, liver fibrosis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), liver cirrhosis, alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), HCV-associated cirrhosis, drug-induced liver injury, hepatocellular necrosis, and / or hepatocellular carcinoma.

[00168] “Therapeutically effective quantity”, in this descriptive report, is intended to include the quantity of an RNAi agent that, when Petition 870260042207, dated 05 / 05 / 2026, page 69 / 431 / 386, administered to an individual with a disease, disorder, or condition associated with HSD17B13, is sufficient for the effective treatment of the disease (e.g., decreasing, improving, or maintaining existing disease or one or more disease symptoms). The “therapeutically effective amount” may vary depending on the RNAi agent, how the agent is administered, the disease and its severity, and the history, age, weight, family history, genetic makeup, types of prior and concurrent treatments, if any, and other personal characteristics of the individual being treated.

[00169] “Prophylactically effective amount,” in this descriptive report, is intended to include the amount of an iRNA that, when administered to an individual with a disease, disorder, or condition associated with HSD17B13, is sufficient to prevent or improve the disease or one or more symptoms of the disease. Improving the disease includes slowing the progression of the disease or reducing the severity of disease that develops later. The “prophylactically effective amount” may vary depending on the iRNA, the method of administration, the degree of disease risk, and the individual’s history, age, weight, family history, genetic makeup, prior and concurrent treatments, if any, and other personal characteristics of the individual being treated.

[00170] A “therapeutically effective amount” or “prophylactically effective amount” also includes an amount of an RNAi agent that produces some desired local or systemic effect with a reasonable risk / benefit ratio that can be applied to any treatment. The RNAi employed in the methods of the present invention can be administered in a sufficient amount to produce a reasonable risk / benefit ratio applicable to such treatment.

[00171] The expression “pharmaceutically acceptable” is used here to refer to those compounds, materials, compositions and / or pharmaceutical forms that are, within the scope of reasoned medical judgment, suitable. Petition 870260042207, dated 05 / 05 / 2026, page 70 / 431 / 386 for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response or other problem or complication, commensurate with a reasonable risk / benefit.

[00172] The term “pharmaceutically acceptable vehicle” in this descriptive report means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc, magnesium, calcium, or zinc stearate, or stearic acid), or encapsulation solvent, involved in carrying or transporting the compound in question from one organ or body part to another organ or body part. Each vehicle must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not harmful to the individual being treated. Some examples of materials that may serve as pharmaceutically acceptable vehicles 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; (7) lubricating agents, such as magnesium stearate, sodium lauryl sulfate and talc; (8) excipients, such as cocoa butter and suppository waxes; (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) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline solution; (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 (22) other non-compatible substances; Petition 870260042207, dated 05 / 05 / 2026, page 71 / 431 / 386 toxic substances used in pharmaceutical formulations.

[00173] The term “sample,” in this descriptive report, includes a collection of fluids, cells, or similar tissues isolated from an individual, as well as fluids, cells, or tissues present within an individual. Examples of biological fluids include blood, serum and serous fluids, plasma, cerebrospinal fluid, ocular fluids, lymph, urine, saliva, and the like. Tissue samples may include samples of tissues, organs, or localized regions. For example, samples may be derived from particular organs, parts of organs, or fluids or cells within those organs. In certain embodiments, samples may be derived from the liver (e.g., whole liver or certain segments of the liver or certain types of cells in the liver, such as, e.g., hepatocytes). In some embodiments, a “sample derived from an individual” refers to blood or plasma taken from the individual. II. iRNAs of the invention

[00174] iRNAs that inhibit the expression of a target gene are described herein. In one embodiment, the iRNAs inhibit the expression of an HSD17B13 gene. In one embodiment, the iRNA agent includes double-stranded ribonucleic acid (dsRNA) molecules to inhibit the expression of an HSD17B13 gene in a cell, such as a liver cell, in an individual, e.g., a mammal, such as a human with a chronic fibro-inflammatory liver disease, disorder, or condition, e.g., a disease, disorder, or condition associated with, e.g., the accumulation and / or expansion of lipid droplets in the liver and / or liver fibrosis.

[00175] The dsRNA includes an antisense strand with a complementarity region that is complementary to at least part of an mRNA formed in the expression of an HSD17B13 gene. The complementarity region is approximately 30 or fewer nucleotides in length (e.g., approximately 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, Petition 870260042207, dated 05 / 05 / 2026, page 72 / 431 / 386 20, 19, or 18 or fewer nucleotides in length). Upon contact with a cell expressing the target gene, the iRNA inhibits the expression of the target gene (e.g., a human, primate, non-primate, or avian target gene) by at least approximately 10%, as assessed, for example, by a PCR-based or branched DNA (bDNA)-based method, or by a protein-based method, such as immunofluorescence analysis, using, for example, Western blotting or flow cytometry techniques.

[00176] A dsRNA includes two RNA strands that are complementary and hybridize to form a duplex structure under conditions in which the dsRNA will be used. One strand of a dsRNA (the antisense strand) includes a region of complementarity that is substantially complementary and, in general, fully complementary to a target sequence. The target sequence may be derived from the sequence of an mRNA formed during the expression of an HSD17B13 gene. The other strand (the sense strand) includes a region that is complementary to the antisense strand, so that the two strands hybridize and form a duplex structure when combined under suitable conditions. As described elsewhere herein and as known in the art, the complementary sequences of a dsRNA may also be contained as self-complementary regions of a single nucleic acid molecule, as opposed to being separate oligonucleotides.

[00177] In general, the duplex structure has between 15 and 30 base pairs in length, p. e.g., between 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 pairs of base lengths. Intermediate intervals and lengths to the intervals and lengths mentioned above are also contemplated as part of the invention. Petition 870260042207, dated 05 / 05 / 2026, page 73 / 431 / 386

[00178] Similarly, the region of complementarity with the target sequence is between 15 and 30 nucleotides long, e.g., between 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 nucleotide lengths. Intermediate intervals and lengths to the intervals and lengths mentioned above are also contemplated as part of the invention.

[00179] In some embodiments, the sense and antisense strands of dsRNA are independently between approximately 15 and 30 nucleotides in length, or between approximately 25 and 30 nucleotides in length, p. For example, each tape independently has between 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. In some embodiments, dsRNA is between approximately 15 and 23 nucleotides in length, or between approximately 25 and 30 nucleotides in length. In general, dsRNA is long enough to serve as a substrate for the Dicer enzyme.For example, it is well known in the art that dsRNAs longer than 21-23 nucleotides can serve as substrates for Dicer. As will also be recognized by any person skilled in the art, the region of an RNA targeted for cleavage will most often be part of a larger RNA molecule, often an mRNA molecule. When relevant, a “part” of a target mRNA is a contiguous sequence of a target mRNA. Petition 870260042207, dated 05 / 05 / 2026, p. 74 / 431 / 386 of sufficient length to allow it to be a substrate for RNAi-directed cleavage (i.e., cleavage via a RISC pathway).

[00180] Those skilled in the art will also recognize that the duplex region is a primary functional portion of a dsRNA, e.g., a duplex region with approximately 9 to 36 base pairs, p. e.g., approximately 10-36, 11-36, 12-36, 13-36, 14-36, 15-36, 9-35, 10-35, 11-35, 12-35, 13-35, 14-35, 15-35, 9-34, 10-34, 11-34, 12-34, 13-34, 14-34, 15-34, 9-33, 10-33, 11-33, 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, 19-20, 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. Thus, in one embodiment, to the extent that it becomes processed into a functional duplex of, e.g., 15-30 base pairs, which directs a desired RNA for cleavage, an RNA molecule or complex of RNA molecules with a duplex region larger than 30 base pairs is a dsRNA. Thus, any person skilled in the art will recognize that, in one embodiment, a miRNA is a dsRNA. In another embodiment, a dsRNA is not a naturally occurring miRNA. In yet another embodiment, an iRNA agent useful for targeting HSD17B13 expression is not generated in the target cell by cleavage of a larger dsRNA.

[00181] A dsRNA as described herein may also include one or more single-stranded nucleotide overhangs, e.g., 1, 2, 3, or 4 nucleotides. dsRNAs with at least one nucleotide overhang may exhibit unexpectedly superior inhibitory properties compared to their blunt-ended counterparts. A nucleotide overhang may comprise or consist of a nucleotide / nucleoside analog, Petition 870260042207, dated 05 / 05 / 2026, page 75 / 431 / 386 including a deoxynucleotide / nucleoside. The overhang(s) may be on the sense strand, the antisense strand, or any combination thereof. Furthermore, the nucleotide(s) of an overhang may be present at the 5' end, the 3' end, or both ends of an antisense or sense strand of a dsRNA.

[00182] A dsRNA can be synthesized by standard methods known in the art, as discussed in more detail below, e.g., using an automated DNA synthesizer, such as those commercially available, for example, from Biosearch, Applied Biosystems, Inc.

[00183] The iRNA compounds of the invention can be prepared by means of a two-step procedure. First, the individual strands of the double-stranded RNA molecule are prepared separately. Then, the component strands are annealed. The individual strands of the siRNA compound can be prepared by means of organic synthesis in soluble phase or solid phase, or both. Organic synthesis offers the advantage that oligonucleotide strands comprising non-natural or modified nucleotides can be easily prepared. The single-stranded oligonucleotides of the invention can be prepared by means of organic synthesis in soluble phase or solid phase, or both.

[00184] In one aspect, a dsRNA of the invention includes at least two nucleotide sequences, a sense sequence and an antisense sequence. The sense strand sequence is selected from the group of sequences provided in any of Tables 2, 3, 7, 8, 10, 11, or 13, and the antisense strand nucleotide sequence, corresponding to the sense strand, is selected from the group of sequences in any of Tables 2, 3, 7, 8, 10, 11, or 13. In this aspect, one of the two sequences is complementary to the other of the two sequences, with one of the sequences being substantially complementary to a sequence of an mRNA generated in the expression of a Petition 870260042207, dated 05 / 05 / 2026, p. 76 / 431 / 386 gene HSD17B13. Thus, in this respect, a dsRNA will include two oligonucleotides, wherein one oligonucleotide is described as the sense strand (passenger strand) in any of Tables 2, 3, 7, 8, 10, 11 or 13, and the second oligonucleotide is described as the corresponding antisense strand (guide strand) of the sense strand in any of Tables 2, 3, 7, 8, 10, 11 or 13. In one embodiment, substantially complementary dsRNA sequences are contained in separate oligonucleotides. In another embodiment, the substantially complementary dsRNA sequences are contained in a single oligonucleotide.

[00185] It will be understood that, although the sequences in Tables 2, 3, 7, 8, 10, 11 or 13 are described as modified, unmodified non-conjugated and / or conjugated sequences, the RNA of the iRNA of the invention, e.g., a dsRNA of the invention, may comprise any of the sequences presented in any of Tables 2, 3, 7, 8, 10, 11 or 13 that is unmodified, non-conjugated and / or modified and / or conjugated in a manner different from that described therein.

[00186] Those skilled in the art are well aware that dsRNAs with a duplex structure between approximately 20 and 23 base pairs, e.g., 21 base pairs, have been noted as particularly effective in inducing RNA interference (Elbashir et al., (2001) EMBO J., 20:6877-6888). However, others have 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 embodiments described above, by virtue of the nature of the oligonucleotide sequences provided herein, the dsRNAs described herein may include at least one strand with a length of at least 21 nucleotides. It can reasonably be predicted that shorter duplexes, with a few fewer nucleotides at one or both ends, may be equally effective when compared to the dsRNAs described above. Consequently, dsRNAs with a sequence of Petition 870260042207, dated 05 / 05 / 2026, p. 77 / 431 / 386 at least 15, 16, 17, 18, 19, 20 or more contiguous nucleotides, derived from one of the sequences provided herein, and differing in the ability to inhibit the expression of an HSD17B13 gene by no more than approximately 5, 10, 15, 20, 25 or 30% of the inhibition of a dsRNA comprising the complete sequence, are contemplated and covered by the scope of the present invention.

[00187] In addition, the RNAs described in any of Tables 2, 3, 7, 8, 10, 11, or 13 identify one or more sites in an HSD17B13 transcript that are susceptible to RISC-mediated cleavage. Thus, the present invention also provides iRNAs that target an iRNA within that site(s). In this descriptive report, an iRNA is said to target an iRNA within a particular site of an RNA transcript if the iRNA promotes transcript cleavage anywhere within that specific site.Such iRNA will generally include at least approximately 15 contiguous nucleotides from one of the sequences provided here, attached to additional nucleotide sequences taken from the region contiguous to the selected sequence in the gene.

[00188] Although a target sequence is generally approximately 15-30 nucleotides long, there is considerable variation in the suitability of specific sequences within this range for directing the cleavage of any given target RNA. Several software packages and the guidelines presented here provide guidance for identifying optimal target sequences for any given target gene, but an empirical approach can also be adopted, in which a “window” or “mask” of a given size (as a non-limiting example, 21 nucleotides) is literally or figuratively (including, e.g., in silico) placed on the target RNA sequence to identify sequences within the size range that could serve as target sequences. By progressively shifting the sequence “window” one nucleotide upstream or downstream from an initial target sequence location, the next potential target sequence can be identified, until the set Petition 870260042207, dated 05 / 05 / 2026, p. 78 / 431 / 386 complete set of possible sequences is identified for any given selected target size. This process, coupled with the systematic synthesis and testing of the identified sequences (through assays as described herein or as known in the art) to identify those sequences with optimal performance, can identify those RNA sequences that, when targeted with an iRNA agent, mediate the best inhibition of target gene expression. Thus, although the sequences identified herein represent effective target sequences, it is contemplated that further optimization of inhibition efficiency can be achieved by “shifting the window” progressively by one nucleotide upstream or downstream in the given sequences to identify sequences with equal or better inhibition characteristics.

[00189] Furthermore, it is contemplated that, for any sequence identified herein, it would be possible to achieve further optimization by systematically adding or removing nucleotides to generate longer or shorter sequences and testing these generated sequences by shifting a window of the longer or shorter size up or down the target RNA from that point. Again, this approach to generating new candidate targets, coupled with tests of the effectiveness of iRNAs based on these target sequences in an inhibition assay, as known in the art and / or as described herein, can lead to further improvements in inhibition efficiency. Moreover, such optimized sequences can be adjusted, e.g., by introducing modified nucleotides, as described herein or known in the art, by adding or altering the overhang, or by other modifications known in the art and / or discussed herein to further optimize the molecule (e.g.to increase serum stability or circulating half-life, increase thermal stability, enhance transmembrane delivery, target a particular location or cell type, increase interaction with silencing pathway enzymes, and increase release from endosomes. Petition 870260042207, dated 05 / 05 / 2026, page 79 / 431 / 386 as an expression inhibitor.

[00190] An iRNA agent as described herein may contain one or more mismatches with respect to the target sequence. In one embodiment, an iRNA as described herein contains no more than 3 mismatches. If the antisense strand of the iRNA contains mismatches with respect to a target sequence, it is preferable that the mismatch area not be located in the center of the complementarity region. If the antisense strand of the iRNA contains mismatches with respect to the target sequence, it is preferable that the mismatch be restricted within the last 5 nucleotides from the 5' or 3' end of the complementarity region. For example, for an iRNA agent with 23 nucleotides, the strand that is complementary to a region of an HSD17B13 gene generally does not contain mismatches within the central 13 nucleotides.The methods described herein, or methods known in the art, can be used to determine whether an iRNA containing a mismatch with respect to a target sequence is effective in inhibiting the expression of an HSD17B13 gene. Considering the effectiveness of mismatched iRNAs in inhibiting the expression of an HSD17B13 gene is important, especially if the specific complementarity region in an HSD17B13 gene is known to have polymorphic sequence variation within the population. III. Modified iRNAs of the invention

[00191] In one embodiment, the RNA of the iRNA of the invention (e.g., a dsRNA) is unmodified and does not comprise, for example, chemical modifications and / or conjugations known in the art and described herein. In another embodiment, the RNA of an iRNA of the invention (e.g., a dsRNA) is chemically modified to enhance stability or other beneficial characteristics. In certain embodiments of the invention, substantially all nucleotides of an iRNA of the invention are modified. In other embodiments of the invention, all nucleotides of Petition 870260042207, dated 05 / 05 / 2026, p. 80 / 431 / 386, an iRNA of the invention is modified. iRNAs of the invention in which “substantially all nucleotides are modified” are modified to a great extent, but not entirely, and do not include more than 5, 4, 3, 2, or 1 unmodified nucleotides.

[00192] In some aspects of the invention, substantially all nucleotides of an iRNA of the invention are modified and the iRNA agents comprise no more than 10 nucleotides comprising 2'-fluoro modifications (e.g., no more than 9 2'-fluoro modifications, no more than 8 2'-fluoro modifications, no more than 7 2'-fluoro modifications, no more than 6 2'-fluoro modifications, no more than 5 2'-fluoro modifications, no more than 4 2'-fluoro modifications, no more than 5 2'-fluoro modifications, no more than 4 2'-fluoro modifications, no more than 3 2'-fluoro modifications or no more than 2'-fluoro modifications). For example, in some embodiments, the sense strand comprises no more than 4 nucleotides comprising 2'-fluoro modifications (e.g., no more than 3 2'-fluoro modifications or no more than 2 2'-fluoro modifications). In other embodiments, the antisense strand comprises no more than 6 nucleotides comprising 2'-fluoro modifications (e.g.(no more than 5 2'-fluoro modifications, no more than 4 2'-fluoro modifications, no more than 4 2'-fluoro modifications, or no more than 2 2'-fluoro modifications).

[00193] In other aspects of the invention, all nucleotides of an iRNA of the invention are modified and the iRNA agents comprise no more than 10 nucleotides comprising 2'-fluoro modifications (e.g., no more than 9 2'-fluoro modifications, no more than 8 2'-fluoro modifications, no more than 7 2'-fluoro modifications, no more than 6 2'-fluoro modifications, no more than 5 2'-fluoro modifications, no more than 4 2'-fluoro modifications, no more than 5 2'-fluoro modifications, no more than 4 2'-fluoro modifications, no more than 3 2'-fluoro modifications or no more than 2 2'-fluoro modifications).

[00194] In one embodiment, the double-stranded RNAi agent of the invention Petition 870260042207, dated 05 / 05 / 2026, page 81 / 431 / 386, further comprises a 5'-phosphate or a 5'-phosphate mimetic at the 5' nucleotide of the antisense strand. In another embodiment, the double-stranded RNAi agent further comprises a 5'-phosphate mimetic at the 5' nucleotide of the antisense strand. In a specific embodiment, the 5'-phosphate mimetic is 5'-vinylphosphate (5'-VP).

[00195] The nucleic acids presented in the invention can be synthesized and / or modified by methods well established in the art, such as those described in “Current protocols in nucleic acid chemistry”, Beaucage, SL et al. (Edrs.), John Wiley & Sons, Inc., New York, NY, USA, the content of which is incorporated herein by reference. Modifications include, for example, end modifications, e.g., 5' end modifications (phosphorylation, conjugation, inverted linkages) or 3' end modifications (conjugation, DNA nucleotides, inverted linkages, etc.); base modifications, e.g., substitution with stabilizing bases, destabilizing bases, or bases that pair the base 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 substitution; and / or modifications to the backbone, including modification or substitution of phosphodiester bonds. Specific examples of useful iRNA compounds in the embodiments described herein include, but are not limited to, RNAs containing modified backbones or lacking natural nucleotide linkages. RNAs with modified backbones include, but are not limited to, those lacking a phosphorus atom in the backbone. For the purposes of this descriptive report, and as sometimes mentioned in the art, modified RNAs lacking a phosphorus atom in their internucleosidic backbone may also be considered oligonucleosides. In some embodiments, a modified iRNA will have a phosphorus atom in its internucleosidic backbone. Petition 870260042207, dated 05 / 05 / 2026, page 82 / 431 / 386

[00196] Modified RNA backbones include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters, methyl and other alkyl phosphonates including 3'-alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates including 3'-aminophosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters and boranephosphates with normal 3'-5' linkages, 2'-5'-linked analogs thereof and those with inverted polarity, in which the pairs adjacent to the 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 invention, the dsRNA agents of the invention are in the form of free acid. In other embodiments of the invention, the dsRNA agents of the invention are in salt form. In one embodiment, the dsRNA agents of the invention are in a sodium salt form.In certain embodiments, when the dsRNA agents of the invention are in the form of a sodium salt, sodium ions are present in the agent as counterions of substantially all phosphodiester and / or phosphorothioate groups present in the agent. Agents in which substantially all phosphodiester and / or phosphorothioate linkages have a sodium counterion do not include more than 5, 4, 3, 2, or 1 phosphodiester and / or phosphorothioate linkages without a sodium counterion. In some embodiments, when the dsRNA agents of the invention are in the form of a sodium salt, sodium ions are present in the agent as counterions of all phosphodiester and / or phosphorothioate groups present in the agent.

[00197] Representative US patents teaching the preparation of the above phosphorus-containing bonds include, among others, US Patents 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; 5,286,717; 5,321,131; 5,399,676; 5,405,939; 5,453,496; 5,455,233; 5,466,677; 5,476,925; 5,519,126; 5,536,821; 5,541,316; 5,550,111; 5,563,253; 5,571,799; 5,587,361; 5,625,050; 6,028,188; 6,124,445; Petition 870260042207, dated 05 / 05 / 2026, page 83 / 431 / 386 160 109; 6 169 170; 6 172 209; 6 239 265; 6 277 603; 6 326 199; 6 346 614; 6 444 423; 6 531 590; 6 534 639; 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 US Patent RE39464, the contents of which are incorporated herein in their entirety by reference.

[00198] Modified RNA backbones that do not include a phosphorus atom have backbones that are formed by short-chain alkyl or cycloalkyl linkages between nucleosides, mixed heteroatoms and alkyl and cycloalkyl linkages between nucleosides, or one or more short-chain heteroatom or heterocyclic linkages between nucleosides. These include those with morpholino linkages (formed in part from the sugar portion of a nucleoside); siloxane backbones; sulfide, sulfoxide, and sulfone backbones; formacetyl and thioformacetyl backbones; methylene formacetyl and thioformacetyl backbones; alkene-containing backbones; sulfamate backbones; methyleneimino and methylenehydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and others having mixed component parts of N, O, S and CH2.

[00199] Representative US patents teaching the preparation of the above oligonucleosides include, among others, US Patents 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; 5,489,677; 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 contents of which are incorporated herein in their entirety by reference.

[00200] In other embodiments, suitable RNA mimetics are contemplated for use in iRNAs, in which both the sugar and the linkage between nucleosides, i.e., the main chain of nucleotide units, are replaced by new groups. The base units are Petition 870260042207, dated 05 / 05 / 2026, page 84 / 431 / 386 maintained for hybridization with an appropriate nucleic acid target compound. One such oligomeric compound, an RNA mimetic that has demonstrated excellent hybridization properties, is referred to as peptidonucleic acid (PNA). In PNA compounds, the acidic backbone of an RNA is replaced by an amide-containing backbone, especially an aminoethylglycine backbone. The nucleobases are retained and linked directly or indirectly to aza nitrogen atoms of the amide portion of the backbone. Representative US patents teaching the preparation of PNA compounds include, among others, US Patents Nos. 5,539,082; 5,714,331; and 5,719,262, the contents of which are incorporated herein in their entirety by reference. Additional PNA compounds for use in the iRNAs of the invention are described in, for example, Nielsen et al., Science, 1991, 254, 1497-1500.

[00201] Some embodiments presented in the invention include RNAs with phosphorothioate backbones and oligonucleosides with backbones containing heteroatoms and, in particular, --CH2--NH-CH2-, --CH2--N(CH3)--O--CH2--[known as methylene (methylimino) or MMI backbone], --CH2--O--N(CH3)--CH2--, --CH2--N(CH3)-N(CH3)--CH2-- and --N(CH3)--CH2 [wherein the native phosphodiester backbone is represented as --O--P--O--CH2--] of U.S. Patent No. 489677 mentioned above, and the amide backbones of U.S. Patent No. 602240 mentioned above. In some embodiments, the RNAs presented here possess back-strand structures with morpholino as per U.S. Patent No. 5,034,506 mentioned above.

[00202] Modified RNAs may also contain one or more substituted sugar moieties. iRNAs, for example (e.g., dsRNAs, presented herein), may include one of the following at the 2' position: OH; F; O-, S- or N-alkyl; O-, S- or N-alkenyl; O-, S- or N-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl, alkenyl and C10 alkyl may be substituted or unsubstituted. Petition 870260042207, dated 05 / 05 / 2026, p. 85 / 431 / 386 or alkenyl and alkynyl C2 to C10. Suitable exemplary modifications include O[(CH2)nO]mCH3, O(CH2).nOCH3, O(CH2)nNH2, O(CH2)nCH3, O(CH2)nONH2, and O(CH2)nON[(CH2)nCH3)]2, where n are from 1 to approximately 10. In other embodiments, dsRNAs include one of the following at the 2' position: C1 to C10 lower alkyl, substituted lower alkyl, alkylaryl, arylalkyl, O-alkylaryl or O-arylalkyl, SH, SCH3, OCN, Cl, Br, CN, CF3, OCF3, SOCH3, SO2CH3, ONO2, NO2, N3, NH2, heterocycloalkyl, heterocycloalkylaryl, aminoalkylamino, polyalkylamino, substituted silyl, a group for RNA cleavage, a reporter group, an intercalator, a group to improve properties pharmacokinetics of an iRNA or a group to improve the pharmacodynamic properties of an iRNA, and other substituents with similar properties.In some embodiments, the modification includes a 2'-methoxyethoxy (2'-O--CH2CH2OCH3, 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 2'-dimethylamino-oxyethoxy, i.e., an O(CH2)2ON(CH3)2 group, also known as 2'-DMAOE, as described 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. Additional exemplary modifications include: 5'-Me2'-F nucleotides, 5'-Me-2'-OMe nucleotides, 5'-Me-2'-deoxynucleotides (R and S isomers in these three families); 2'-alkoxyalkyl; and 2'-NMA (N-methylacetamide).

[00203] Other modifications include 2'-methoxy (2'-OCH3), 2'-aminopropoxy (2'-OCH2CH2CH2NH2), and 2'-fluoro (2'-F). Similar modifications can also be made at other positions in the RNA of an iRNA, especially the 3' position of the sugar in the 3'-terminal nucleotide or in 2'-5' linked dsRNAs and at the 5' position of the 5' nucleotide. iRNAs can also have sugar mimetics, such as cyclobutyl moieties in place of the pentofuranosyl sugar. Representative US patents that teach the Petition 870260042207, dated 05 / 05 / 2026, page 86 / 431 / 386 preparation of such modified sugar structures includes, among others, US Patents 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; 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 common property with the present patent application. The content of each of the foregoing is incorporated herein in its entirety by reference.

[00204] An iRNA of the invention may also include modifications or substitutions of the nucleobase (often referred to in the art simply as “base”). In this descriptive report, “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 synthetic and other 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-propynyl-uracil and cytosine, 6-azo-uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkylam 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo, especially 5-bromo, 5-trifluoromethyl and Other uracils and 5-substituted cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-daazaadenine, and 3-deazaguanine and 3-deazaadenine. Additional nucleobases include those disclosed in U.S. Patent No. 3,687,808, and those disclosed in Modified Nucleosides in Biochemistry, Biotechnology and Medicine, Herdewijn, P. ed.Wiley-VCH, 2008; those revealed in The Concise Encyclopedia Of Polymer Science and Engineering, pages 858-859, Kroschwitz, J. L, ed. John Wiley & Sons, 1990, those disclosed by Englisch et al., (1991) Angewandte Chemie, International Edition, 30:613, and those disclosed by. Petition 870260042207, dated 05 / 05 / 2026, p. 87 / 431 / 386 Sanghvi, Y S., Chapter 15, dsRNA Research and Applications, pages 289-302, Crooke, ST and Lebleu, B., Ed., CRC Press, 1993. Some of these nucleobases are especially useful for increasing the binding affinity of the oligomeric compounds presented in the invention. These include 5-substituted pyrimidines, 6-azapyrimidines, and N-2, N-6, and O-6 substituted purines, including 2-aminopropyladenine, 5-propynyluracil, and 5-propynylcytosine. 5-Methylcytosine substitutions have been shown to increase the stability of the nucleic acid duplex at 0.6-1.2 °C (Sanghvi, YS, Crooke, ST and Lebleu, B., Eds., dsRNA Research and Applications, CRC Press, Boca Raton, 1993, p. 276-278) and are exemplary base substitutions, even more so when combined with 2'-O-methoxyethyl modifications in the sugar.

[00205] Representative US patents teaching the preparation of some of the modified nucleobases noted above, as well as other modified nucleobases, include, among others, US Patents 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; 5,594,121; 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 noted above, the contents of which are incorporated herein in their entirety by reference.

[00206] An iRNA of the invention can also be modified to include one or more locked nucleic acids (LNAs). A locked nucleic acid is a nucleotide with a modified ribose moiety, wherein the ribose moiety comprises an extra bridge connecting the 2' and 4' carbons. This structure effectively “locks” the ribose in the 3' end structural conformation. The addition of locked nucleic acids to siRNAs has been shown to increase siRNA stability in serum and reduce off-target effects (Elmen, J. et al., (2005) Nucleic Acids Research Petition 870260042207, dated 05 / 05 / 2026, page 88 / 431 / 386 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).

[00207] An iRNA of the invention may also be modified to include one or more bicyclic sugar moieties. A “bicyclic sugar” is a furanosyl ring modified by bridging between two atoms. A “bicyclic nucleoside” (“BNA”) is a nucleoside with a sugar moiety comprising a bridge connecting two carbon atoms of the sugar ring, thus forming a bicyclic ring system. In certain embodiments, the bridge connects carbon 4' and carbon 2' of the sugar ring. Thus, in some embodiments an agent of the invention may include one or more blocked nucleic acids (LNAs). A blocked nucleic acid is a nucleotide with a modified ribose moiety, wherein the ribose moiety comprises an extra bridge connecting carbons 2' and 4'. In other words, an LNA is a nucleotide comprising a bicyclic sugar moiety comprising a 4'-CH2-O-2' bridge. This structure effectively "locks" the ribose into the 3' end structural conformation.The addition of blocked nucleic acids to siRNAs 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 the polynucleotides of the invention include, among others, nucleosides comprising a bridge between the 4' and 2' atoms of the ribosyl ring. In certain embodiments, the antisense polynucleotide agents of the invention include one or more bicyclic nucleosides comprising a 4' to 2' bridge. Examples of such bicyclic nucleosides with a 4' to 2' bridge include, among others, 4'-(CH2)-O-2' (LNA); 4'-(CH2)-S-2'; 4'-(CH2)2-O-2' (ENA); 4'-CH(CH3)-O-2' (also referred to as “hindered ethyl” or “cEt”) and 4'CH(CH2OCH3)-O-2' (and analogues thereof; see, e.g., US Patent No. 7). Petition 870260042207, dated 05 / 05 / 2026, p. 89 / 431 / 386 399 845); 4'-C(CH3)(CH3)-O-2' (and analogues thereof; see, e.g., U.S. Patent No. 8,278,283); 4'-CH2-N(OCH3)-2' (and analogues thereof; see, e.g., U.S. Patent 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', where R is H, C1-C12 alkyl or a protecting group (see, for example, U.S. Patent 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 analogues thereof; see, for example, U.S. Patent No. 8,278,426). The contents of each of the foregoing are incorporated herein in their entirety by reference.

[00208] Additional representative US patents and US patent publications teaching the preparation of nucleotides from blocked nucleic acids include, among others, the following: US Patents 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; 7,399,845; 7,427,672; 7,569,686; 7,741,457; 8,022,193; 8,030,467; 8,278,425; 8,278,426; 8,278,283; US 2008 / 0039618 and US 2009 / 0012281, the contents of which are incorporated herein in their entirety by reference.

[00209] Any of the above bicyclic nucleosides can be prepared to have one or more additional stereochemical sugar configurations including, for example, α-L-ribofuranose and β-D-ribofuranose (see WO 99 / 14226).

[00210] An iRNA of the invention can also be modified to include one or more ethyl-hindered nucleotides. In this descriptive report, an “ethyl-hindered nucleotide” or “cEt” is a blocked nucleic acid comprising a bicyclic sugar moiety comprising a 4'-CH(CH3)-O-2' bridge. In one embodiment, an ethyl-hindered nucleotide is in the S conformation, referred to herein as “ScEt”.

[00211] An iRNA of the invention may also include one or more “conformationally restricted nucleotides” (“CRNs”). CRNs are analogs of Petition 870260042207, dated 05 / 05 / 2026, page 90 / 431 / 386 nucleotides with a linker connecting the C2' and C4' carbons of ribose or the C3 and C5' carbons of ribose. CRNs lock the ribose ring in a stable conformation and increase the affinity for hybridization with mRNA. The linker is long enough to position the oxygen in an optimal position for stability and affinity, resulting in less puckering of the ribose ring.

[00212] Representative publications that teach the preparation of some of the CRNs listed above include, among others, US Patent Publication No. 2013 / 0190383; and PCT Publication WO 2013 / 036868, the contents of which are incorporated herein in their entirety by reference.

[00213] In some embodiments, an iRNA of the invention comprises one or more monomers that are UNA nucleotides (unblocking nucleic acid). UNA is an unblocking acyclic nucleic acid in which any of the sugar linkages have been removed, forming an unblocking “sugar” residue. In one example, UNA also encompasses the monomer with C1'-C4' linkages that have been removed (i.e., the carbon-oxygen-carbon covalent bond between carbons C1' and C4'). In another example, the C2'-C3' linkage (i.e., the carbon-carbon covalent bond between carbons C2' and C3') of the sugar has been removed (see, Nuc. Acids Symp. Series, 52, 133-134 (2008) and Fluiter et al., Mol. Biosyst., 2009, 10, 1039, incorporated herein by reference).

[00214] Representative US publications that teach the preparation of UNA include, among others, US Patent No. 8,314,227; and US Patent Publications Nos. 2013 / 0096289; 2013 / 0011922; and 2011 / 0313020, the contents of which are incorporated herein in their entirety by reference.

[00215] Potentially stabilizing modifications to the ends of RNA molecules may include N-(acetylaminocaproyl)-4-hydroxyprolinol (Hyp-C6-NHAc), N-(caproyl-4-hydroxyprolinol (Hyp-C6), N(acetyl-4-hydroxyprolinol (Hyp-NHAc), thymidine-2'-O-deoxythymidine (ether), N-(aminocaproyl)-4-hydroxyprolinol (Hyp-C6-amino), 2-docosanoyl-uridine-3”phosphate, dT inverted base (idT), and others. A description of this modification can be found in PCT Publication No. 870260042207, dated 05 / 05 / 2026, page 91 / 431 / 386.

[00216] Other modifications of an iRNA of the invention include a 5' phosphate or 5' phosphate mimetic, e.g., a 5'-terminal phosphate or phosphate mimetic on the antisense strand of an RNAi agent. Phosphate mimetics are described in, for example, U.S. Patent Publication No. 2012 / 0157511, the contents of which are incorporated herein in their entirety by reference.

[00217] In certain specific embodiments, an RNAi agent of the present invention is an agent that inhibits the expression of an HSD17B13 gene that is selected from the group of agents listed in any of Tables 2, 3, 7, 8, 10, 11, or 13. Any of these agents may further comprise a linker. A. Modified iRNAs comprising reasons for the invention

[00218] In certain aspects of the invention, the double-stranded RNAi agents of the invention include agents with chemical modifications as disclosed, for example, in WO 2013 / 075035, filed on November 16, 2012, the contents of which are incorporated herein in their entirety by reference.

[00219] Consequently, the invention provides double-stranded RNAi agents capable of inhibiting the expression of a target gene (i.e., an HSD17B13 gene) in vivo. The RNAi agent comprises a sense strand and an antisense strand. Each strand of the RNAi agent can vary from 12-30 nucleotides in length. For example, each strand can be between 14-30 nucleotides long, 17-30 nucleotides long, 27-30 nucleotides long, 17-21 nucleotides long, 19-25 nucleotides long, 19-21 nucleotides long, 25-30 nucleotides long, 17-23 nucleotides long, 17-19 nucleotides long, 19-23 nucleotides long, 21-25 nucleotides long, or 21-23 nucleotides long. In one embodiment, the Petition 870260042207, dated 05 / 05 / 2026, page 92 / 431 / 386: The sense strand is 21 nucleotides long. In one embodiment, the antisense strand is 23 nucleotides long.

[00220] The sense strand and the antisense strand typically form a double-stranded duplex RNA (“dsRNA”), also referred to here as an “RNAi agent”. The duplex region of an RNAi agent can be 12-30 nucleotide pairs long. For example, the duplex region can be between 14-30 nucleotide pairs long, 17-30 nucleotide pairs long, 27-30 nucleotide pairs long, 17-23 nucleotide pairs long, 17-21 nucleotide pairs long, 17-19 nucleotide pairs long, 19-25 nucleotide pairs long, 19-23 nucleotide pairs long, 19-21 nucleotide pairs long, 21-25 nucleotide pairs long, or 21-23 nucleotide pairs long. In another example, the duplex region is selected from among 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, and 27 nucleotides in length.

[00221] In one embodiment, the RNAi agent may contain one or more overhang regions and / or capping regions at the 3' end, 5' end, or both ends of one or both strands. The overhang may be 1-6 nucleotides long, for example, 2-6 nucleotides long, 1-5 nucleotides long, 2-5 nucleotides long, 1-4 nucleotides long, 2-4 nucleotides long, 1-3 nucleotides long, 2-3 nucleotides long, or 1-2 nucleotides long. The overhangs may result from one strand being longer than the other, or from two strands of the same length being staggered. The protrusion may form a mismatch with the target mRNA, or it may be complementary to the sequences of the target gene, or it may be another sequence. The first and second strands may also be joined, e.g., by additional bases to form a hairpin. Petition 870260042207, dated 05 / 05 / 2026, p. 93 / 431 / 386 hair) or by linkers other than bases.

[00222] In one embodiment, the nucleotides in the protruding region of the RNAi agent may each independently be a modified or unmodified nucleotide, including, but not limited to, 2'-modified sugars, such as 2-F, 2'-O-methyl, thymidine (T), 2'-O-methoxyethyl-5-methyluridine (Teo), 2'-O-methoxyethyladenosine (Aeo), 2'-O-methoxyethyl-5-methylcytidine (m5Ceo), and any combinations thereof. For example, TT may be a protruding sequence from either end on either strand. The protrusion may form a mismatch with the target mRNA or may be complementary to the target gene sequences or may be another sequence.

[00223] The 5' or 3' overhangs on the sense strand, antisense strand, or both strands of the RNAi agent can be phosphorylated.In some embodiments, the protruding region(s) contain two nucleotides with a phosphorothioate between the two nucleotides, wherein the two nucleotides may be the same or different. In one embodiment, the protrusion is present at the 3' end of the sense strand, antisense strand, or both strands. In another embodiment, this 3' protrusion is present on the antisense strand. In another embodiment, this 3' protrusion is present on the sense strand.

[00224] The RNAi agent may contain only one overhang, which can enhance the RNAi's interference activity without affecting its overall stability. For example, the single-stranded overhang may be located at the 3'-terminal end of the sense strand or, alternatively, at the 3'-terminal end of the antisense strand. RNAi may 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. In general, the antisense strand of RNAi has a nucleotide overhang at the 3' end, and the 5' end is blunt. Although not intended to be theoretically binding, the asymmetric blunt end at the 5' end of the antisense strand and the overhang at the 3' end of the antisense strand favor guide strand loading in the RISC process. Petition 870260042207, dated 05 / 05 / 2026, page 94 / 431 / 386

[00225] In one embodiment, the RNAi agent is a double-ended bluntmer of 19 nucleotides in length, wherein the sense strand contains at least one motif of three 2'-F modifications on three consecutive nucleotides at positions 7, 8, 9 from the 5' end. The antisense strand contains at least one motif of three 2'-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5' end.

[00226] In another embodiment, the RNAi agent is a double-ended bluntmer of 20 nucleotides in length, wherein the sense strand contains at least one motif of three 2'-F modifications on three consecutive nucleotides at positions 8, 9, 10 from the 5' end. The antisense strand contains at least one motif of three 2'-O-methyl modifications on three consecutive nucleotides at positions 11, 12, and 13 from the 5' end.

[00227] In yet another embodiment, the RNAi bluntmer agent is at the double end of 21 nucleotides in length, wherein the sense strand contains at least one motif of three 2'-F modifications on three consecutive nucleotides at positions 9, 10, 11 from the 5' end. The antisense strand contains at least one motif of three 2'-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5' end.

[00228] 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 of three 2'-F modifications on three consecutive nucleotides at positions 9, 10, 11 from the 5' end; the antisense strand contains at least one motif of three 2'-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5' end, wherein one end of the RNAi agent is blunt, while the other end comprises a 2-nucleotide overhang. Preferably, the 2-nucleotide overhang is at the 3' end of the antisense strand. Petition 870260042207, dated 05 / 05 / 2026, page 95 / 431 / 386

[00229] When the 2-nucleotide overhang is at the 3' end of the antisense strand, there may be two phosphorothioate linkages between the three terminal nucleotides, wherein two of the three nucleotides are the overhanging nucleotides, and the third nucleotide is a paired nucleotide next to the overhanging nucleotide. In one embodiment, the RNAi agent additionally has two phosphorothioate linkages between the three terminal nucleotides at the 5' end of the sense strand and at the 5' end of the antisense strand. In one embodiment, every nucleotide in the sense strand and in the antisense strand of the RNAi agent, including the nucleotides that are part of the motifs, are modified nucleotides. In one embodiment, each residue is independently modified with 2'-O-methyl or 3'-fluoro, e.g., in an alternating motif. Optionally, the RNAi agent also includes a linker (preferably GalNAc3).

[00230] In one embodiment, the RNAi agent comprises a sense strand and an antisense strand, wherein the sense strand is 25-30 nucleotide residues long, wherein from the 5' terminal nucleotide (position 1), positions 1 to 23 of the first strand comprise at least 8 ribonucleotides; the antisense strand is 36-66 nucleotide residues long and, from the 3' terminal nucleotide, comprises at least 8 ribonucleotides in positions paired with positions 1-23 of the sense strand to form a duplex; wherein at least the 3' terminal nucleotide of the antisense strand is not paired with the sense strand, and up to 6 consecutive 3' terminal nucleotides are not paired with the sense strand, thus forming a 3' single-stranded overhang with 1-6 nucleotides; wherein the 5' end of the antisense strand comprises 10-30 consecutive nucleotides that are not paired with the sense strand, thus forming a 5' single-stranded overhang of 10-30 nucleotides;whereby at least the 5'-terminal and 3'-terminal nucleotides of the sense strand have their bases paired with nucleotides of the antisense strand when the sense and antisense strands are aligned for complementarity; Petition 870260042207, dated 05 / 05 / 2026, page 96 / 431 / 386 maximum, thus forming a substantially duplex region between the sense and antisense strands; and the antisense strand is sufficiently complementary to a target RNA along at least 19 ribonucleotides of the antisense strand's length to reduce the expression of the target gene when the double-stranded nucleic acid is introduced into a mammalian cell; and wherein the sense strand contains at least one motif of three 2'-F modifications in three consecutive nucleotides, wherein at least one of the motifs occurs near or at the cleavage site. The antisense strand contains at least one motif of three 2'-O-methyl modifications in three consecutive nucleotides near or at the cleavage site.

[00231] In one embodiment, the RNAi agent comprises sense and antisense strands, wherein the RNAi agent comprises a first strand having a length that is at least 25 and at most 29 nucleotides, and a second strand having a length that is at most 30 nucleotides with at least one motif of three 2'-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5' end;wherein the 3' end of the first strand and the 5' end of the second strand form a blunt end, and the second strand is 1-4 nucleotides longer at its 3' end than the first strand, wherein the duplex region is at least 25 nucleotides long, and the second strand is sufficiently complementary to a target mRNA along at least 19 nucleotides of the second strand to reduce the expression of the target gene when the RNAi agent is introduced into a mammalian cell, wherein cleavage by the dicer enzyme of the RNAi agent preferentially results in an siRNA comprising the 3' end of the second strand, thus reducing the expression of the target gene in the mammal. Optionally, the RNAi agent further comprises a linker.

[00232] In one embodiment, the sense strand of the RNAi agent contains at least one motif of three identical modifications in three nucleotides. Petition 870260042207, dated 05 / 05 / 2026, page 97 / 431 / 386 consecutive, in which one of the reasons occurs at the cleavage site on the sense tape.

[00233] In one embodiment, the antisense strand of the RNAi agent may also contain at least one motif of three identical modifications on three consecutive nucleotides, wherein one of the motifs occurs near or at the cleavage site on the antisense strand.

[00234] For an RNAi agent having a duplex region 17-23 nucleotides long, the cleavage site of the antisense strand is typically around positions 10, 11, and 12 from the 5' end. Thus, motifs of three identical modifications can occur at positions 9, 10, 11; positions 10, 11, 12; positions 11, 12, 13; positions 12, 13, 14; or positions 13, 14, 15 of the antisense strand, counting from the 1st nucleotide from the 5' end of the antisense strand, or counting from the 1st paired nucleotide within the duplex region from the 5' end of the antisense strand. The cleavage site on the antisense strand can also change according to the length of the RNAi duplex region from the 5' end.

[00235] The sense strand of the RNAi agent may contain at least one motif of three identical modifications on three consecutive nucleotides at the strand's cleavage site; and the antisense strand may have at least one motif of three identical modifications on three consecutive nucleotides near or at the strand's cleavage site. When the sense strand and the antisense strand form a dsRNA duplex, the sense strand and the antisense strand may be aligned in such a way that one motif of the three nucleotides on the sense strand and one motif of the three nucleotides on the antisense strand have at least one nucleotide overlap, that is, at least one of the three nucleotides of the motif on the sense strand forms a base pair with at least one of the three nucleotides of the motif on the antisense strand. Alternatively, at least two nucleotides may overlap or all three nucleotides may overlap. Petition 870260042207, dated 05 / 05 / 2026, page 98 / 431 / 386

[00236] In one embodiment, the sense strand of the RNAi agent may contain more than one motif of three identical modifications at three consecutive nucleotides. The first motif may occur near or at the cleavage site of the strand, and the other motifs may have a wing-shaped modification. The term “wing-shaped modification,” in this descriptive report, refers to a motif occurring elsewhere on the strand that is separated from the motif near or at the cleavage site of the same strand. The wing-shaped modification is adjacent to the first motif or is separated by at least one or more nucleotides. When the motifs are immediately adjacent to each other, then the chemistry of the motifs is distinct from one another, and when the motifs are separated by one or more nucleotides, then the chemistries may be the same or different. Two or more wing-shaped modifications may be present.For example, when two wing-shaped modifications are present, each wing-shaped modification can occur at an extremity relative to the first motif that is near or at the cleavage site, or on either side of the leading motif.

[00237] In the same way as the sense strand, the antisense strand of the RNAi agent can contain more than one motif of three identical modifications on three consecutive nucleotides, with at least one of the motifs occurring near or at the cleavage site of the strand. This antisense strand may also contain one or more wing-shaped modifications in an alignment similar to the wing-shaped modifications that may be present on the sense strand.

[00238] In one embodiment, the wing-shaped modification in the sense strand or antisense strand of the RNAi agent typically does not include the first or two terminal nucleotides at the 3' end, the 5' end, or both ends of the strand.

[00239] In another embodiment, the wing-shaped modification in the sense or antisense strand of the RNAi agent typically does not include the first or two paired nucleotides within the duplex region at the 3' end, in Petition 870260042207, dated 05 / 05 / 2026, page 99 / 431 / 386 end 5' or at both ends of the tape.

[00240] When the sense strand and the antisense strand of the RNAi agent each contain at least one wing-shaped modification, the wing-shaped modifications may be located at the same end of the duplex region and have an overlap of one, two, or three nucleotides.

[00241] When the sense strand and the antisense strand of the RNAi agent each contain two wing-shaped modifications, the sense strand and the antisense strand can be aligned in such a way that the two modifications of each strand are located at one end of the duplex region, having an overlap of one, two, or three nucleotides; two modifications of each strand are located at the other end of the duplex region, having an overlap of one, two, or three nucleotides; two modifications of one strand are located on each side of the leader motif, having an overlap of one, two, or three nucleotides in the duplex region.

[00242] In one embodiment, every nucleotide in the sense and antisense strands of the RNAi agent, including the nucleotides that are part of the motifs, can be modified. Each nucleotide can be modified with the same or different modification, which may include one or more alterations of one or both of the non-linking oxygens of the phosphate and / or of one or more of the linking oxygens of the phosphate; the alteration of a constituent of the ribose sugar, e.g., of the 2' hydroxyl group in the ribose sugar; the massive substitution of the phosphate portion by “dephospho” linkers; the modification or substitution of a natural base; and the substitution or modification of the ribose-phosphate backbone.

[00243] Because nucleic acids are polymers of subunits, many modifications occur at a position that is repeated within a nucleic acid, e.g., a modification of a base moiety or a phosphate moiety, or at a non-binding O of a phosphate moiety. In some cases, the modification will occur at all the positions in question in the nucleic acid, but Petition 870260042207, dated 05 / 05 / 2026, page 100 / 431 / 386 in many cases will not occur. For example, a modification may occur only at a 3' or 5' terminal position, it may occur only in a terminal region, e.g., at a position on a terminal nucleotide or in the last 2, 3, 4, 5 or 10 nucleotides of a strand. A modification may occur in a double-stranded region, a single-stranded region or in both. A modification may occur only in the double-stranded region of an RNA or it may occur only in a single-stranded region of an RNA. For example, a phosphorothioate modification at a non-binding O position may occur only at one or both ends, it may occur only in a terminal region, e.g. For example, it can occur at a position on a terminal nucleotide or in the last 2, 3, 4, 5, or 10 nucleotides of a strand, or it can occur in both double-stranded and single-stranded regions, especially at the terminals. The 5' end or ends may be phosphorylated.

[00244] It may be possible, for example, to increase stability by including particular bases in overhangs, or by including modified nucleotides or nucleotide substitutes in single-stranded overhangs, e.g., in a 5' or 3' overhang, or in both. For example, it may be desirable to include purine nucleotides in overhangs. In some embodiments, all or some of the bases in a 3' or 5' overhang may be modified, e.g., with a modification described herein. Modifications may include, for example, the use of modifications at the 2' position of the ribose sugar with modifications known in the art, e.g., the use of deoxyribonucleotides modified with 2'-deoxy-2'-fluoro (2'-F) or 2'-O-methyl instead of the ribose sugar of the nucleobase, and modifications to the phosphate group, e.g. e.g., phosphorothioate modifications. The overhangs do not need to be homologous with the target sequence.

[00245] In one embodiment, each residue of the 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, Petition 870260042207, dated 05 / 05 / 2026, page 101 / 431 / 386 2'-hydroxyl or 2'-fluoro. The strands may contain more than one modification. In one embodiment, each residue of the sense strand and the antisense strand is independently modified with 2'-O-methyl or 2'-fluoro.

[00246] At least two different modifications are typically present on the sense strand and the antisense strand. These two modifications may be 2'-O-methyl or 2'-fluoro modifications, or others.

[00247] In one embodiment, Na and / or Nb comprise modifications of an alternating pattern. The term “alternating motif,” in this descriptive report, refers to a motif having one or more modifications, each modification occurring on alternating nucleotides of a strand. The alternating nucleotide may refer to one of two nucleotides or one of three nucleotides, or a similar pattern. For example, if A, B, and C each represent one type of nucleotide modification, the alternating motif may be “ABABABABABAB.”, “AABBAABBAABB.”, “AABAABAABAAB.”, “AAABAAABAAAB.”, “AAABBBAAABBB...” or “ABCABCABCABC.” etc.

[00248] The type of modifications contained in the alternating motif can be the same or different. For example, if A, B, C, D each represent a type of modification in the nucleotide, the alternating pattern, that is, modifications every two nucleotides, can be the same, but each of the sense or antisense strands can be selected from several possibilities of modifications within the alternating motif such as “ABABAB...”, “ACACAC...” “BDBDBD.” or “CDCDCD.” etc.

[00249] In one embodiment, the RNAi agent of the invention comprises a change in the modification pattern for the alternating motif on the sense strand relative to the modification pattern for the alternating motif on the antisense strand. The change may be such that the modified group of nucleotides on the sense strand corresponds to a differently modified group of nucleotides on the antisense strand and vice versa. For example, the sense strand when paired Petition 870260042207, dated 05 / 05 / 2026, page 102 / 431 / 386, with the antisense strand in the dsRNA duplex, the alternating motif on the sense strand can start with “ABABAB” from 5'-3' of the strand and the alternating motif on the antisense strand can start with “BABABA” from 5'-3' of the strand within the duplex region. As another example, the alternating motif on the sense strand can start with “AABBAABB” from 5'-3' of the strand and the alternating motif on the antisense strand can start with “BBAABBAA” from 5'-3' of the strand within the duplex region, so there is a complete or partial change of modification patterns between the sense strand and the antisense strand.

[00250] In one embodiment, the RNAi agent comprises the alternating motif pattern of the 2'-O-methyl modification and the 2'-F modification on the sense strand initially has a change from the alternating motif pattern of the 2'-O-methyl modification and the 2'-F modification on the antisense strand initially, i.e., the nucleotide modified with 2'-O-methyl on the sense strand base-pairs with a nucleotide modified with 2'-F on the antisense strand and vice versa. Position 1 of the sense strand can start with the 2'-F modification and position 1 of the antisense strand can start with the 2'-O-methyl modification.

[00251] The introduction of one or more identical three-modification motifs at three consecutive nucleotides on the sense and / or antisense strand disrupts the initial modification pattern present on the sense and / or antisense strand. This disruption of the sense and / or antisense strand modification pattern by the introduction of one or more identical three-modification motifs at three consecutive nucleotides on the sense and / or antisense strand surprisingly intensifies the gene silencing activity of the target gene.

[00252] In one embodiment, when the motif of three identical modifications in three consecutive nucleotides is introduced into either strand, the nucleotide modification following the motif is a different modification from the motif modification. For example, the sequence portion Petition 870260042207, dated 05 / 05 / 2026, page 103 / 431 / 386 containing the motif “.. .NaYYYNb...”, where “Y” represents the modification of the motif of three identical modifications in three consecutive nucleotides, and “Na” and “Nb” represent a modification to the nucleotide following the “YYY” motif that is different from the Y modification, and where Na and Nb may be the same but different modifications. Alternatively, Na and / or Nb may be present or absent when a wing-shaped modification is present.

[00253] The RNAi agent may further comprise at least one phosphorothioate or methylphosphonate linkage between nucleotides. The phosphorothioate or methylphosphonate linkage modification between nucleotides may occur at any nucleotide of the sense strand or the antisense strand or both strands at any position on the strand. For example, the linkage modification between nucleotides may occur at any nucleotide in the sense strand and / or the antisense strand; each linkage modification between nucleotides may occur in an alternating pattern in the sense strand and / or the antisense strand; or the sense strand or the antisense strand may contain both linkage modifications between nucleotides in an alternating pattern.The alternating pattern of nucleotide linkage modification in the sense strand may be the same as or different from that of the antisense strand, and the alternating pattern of nucleotide linkage modification in the sense strand may have a change relative to the alternating pattern of nucleotide linkage modification in the antisense strand. In one embodiment, a double-stranded RNAi agent comprises 6-8 phosphorothioate linkages between nucleotides. In another embodiment, the antisense strand comprises two phosphorothioate linkages between nucleotides at the 5' end and two phosphorothioate linkages between nucleotides at the 3' end, and the sense strand comprises at least two phosphorothioate linkages between nucleotides at either the 5' or 3' end.

[00254] In one embodiment, RNAi comprises a phosphorothioate or methylphosphonate linkage modification between nucleotides in the overhang region. For example, the overhang region may contain two nucleotides. Petition 870260042207, dated 05 / 05 / 2026, page 104 / 431 / 386 having a phosphorothioate or methylphosphonate linkage between the two nucleotides. Modifications to the linkage between nucleotides can also be made to link the protruding nucleotides with the paired terminal nucleotides within the duplex region. For example, at least 2, 3, 4 or all of the nucleotides of the protrusion can be joined via phosphorothioate or methylphosphonate linkage between nucleotides and, optionally, there may be additional phosphorothioate or methylphosphonate linkages between nucleotides linking the protruding nucleotide with a paired nucleotide that is next to the protruding nucleotide. For example, there may be at least two phosphorothioate linkages between the three terminal nucleotides, where two of the three nucleotides are protruding nucleotides, and the third is a nucleotide paired alongside the protruding nucleotide.These three terminal nucleotides can be located at the 3' end of the antisense strand, at the 3' end of the sense strand, at the 5' end of the antisense strand, and / or at the 5' end of the antisense strand.

[00255] In one embodiment, the 2-nucleotide overhang is at the 3' end of the antisense strand, and there are two phosphorothioate linkages between the three terminal nucleotides, wherein two of the three nucleotides are overhang nucleotides, and the third nucleotide is a nucleotide paired alongside the overhang nucleotide. Optionally, the RNAi agent may additionally have two phosphorothioate linkages between the three terminal nucleotides at the 5' end of the sense strand and at the 5' end of the antisense strand.

[00256] In one embodiment, the RNAi agent comprises pairing error(s) with the target, within the duplex, or combinations thereof. The pairing error may occur in the salient region or in the duplex region. The base pair may be classified based on its propensity to promote dissociation or fusion (e.g., in the free energy of association or dissociation of a particular pairing, the simplest approach is to examine pairs pairwise, although nearest neighbor analysis or Petition 870260042207, dated 05 / 05 / 2026, page 105 / 431 / 386 (similar may 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). Pairing errors, e.g., non-canonical or other non-canonical pairings (as described elsewhere in this document) are preferred over canonical pairings (A:T, A:U, G:C); and pairings that include a universal basis are preferred over canonical pairings.

[00257] In one embodiment, the RNAi agent comprises at least one of the first 1, 2, 3, 4, or 5 base pairs within the duplex regions from the 5' end of the antisense strand independently selected from the group of: A:U, G:U, I:C and pairs with mismatch pairings, e.g., non-canonical pairings or other non-canonical pairings or pairings that include a universal base, to promote dissociation of the antisense strand at the 5' end of the duplex.

[00258] In one embodiment, the nucleotide at position 1 within the duplex 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 the first 1, 2, or 3 base pair within the duplex region from the 5' end of the antisense strand is an AU base pair. For example, the first base pair within the duplex region from the 5' end of the antisense strand is an AU base pair.

[00259] In another embodiment, the nucleotide at the 3' end of the sense strand is deoxythymine (dT). In another embodiment, the nucleotide at the 3' end 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.

[00260] In one mode, the sense tape sequence can be represented by formula (I): 5' np-Na-(XXX )i-Nb-YYY-Nb-(ZZZ )J-Na-nq3' (I) Petition 870260042207, dated 05 / 05 / 2026, page 106 / 431 / 386 in which: iej are, each one, independently 0 or 1; small are, each one, independently 0-6; Each Na independently represents an oligonucleotide sequence comprising 0-25 modified nucleotides, each sequence comprising at least two differently modified nucleotides; Each Nb independently represents an oligonucleotide sequence comprising 0-10 modified nucleotides; Each np and nq independently represent a salient nucleotide; where Nb and Y do not have the same modification; and XXX, YYY, and ZZZ each independently represent a motif of three identical modifications in three consecutive nucleotides. Preferably, YYY is all nucleotides modified with 2'-F.

[00261] In one embodiment, Na and / or Nb comprise alternating pattern modifications.

[00262] In one embodiment, the YYY motif occurs near or at the cleavage site of the sense strand. For example, when the RNAi agent has a duplex region of 17-23 nucleotides in length, the YYY motif may occur at or in the vicinity of the cleavage site (e.g., it may occur at positions 6, 7, 8, 7, 8, 9, 8, 9, 10, 9, 10, 11, 10, 11, 12 or 11, 12, 13) of the sense strand, counting from the 1st nucleotide, from the 5' end; or, optionally, counting from the 1st paired nucleotide within the duplex region, from the 5' end.

[00263] In one modality, i is 1 and j is 0, or i is 0 and j is 1, or i and j are both 1. The sense tape can therefore be represented by the following formulas: 5' np-Na-YYY-Nb-ZZZ-Na-nq 3' (Ib); Petition 870260042207, dated 05 / 05 / 2026, page 107 / 431 / 386 5' np-Na-XXX-Nb-YYY-Na-nq 3' (Ic); or 5' np-Na-XXX-Nb-YYY-Nb-ZZZ-Na-nq 3' (Id).

[00264] When the sense strand is represented by the formula (Ib), Nb represents an oligonucleotide sequence comprising 0-10, 0-7, 0-5, 04, 0-2 or 0 modified nucleotides. Each Na can independently represent an oligonucleotide sequence comprising 2-20, 215 or 2-10 modified nucleotides.

[00265] When the sense strand is represented as formula (Ic), Nb represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2, or 0 modified nucleotides. Each Na can independently represent an oligonucleotide sequence comprising 2-20, 215, or 2-10 modified nucleotides.

[00266] When the sense strand is represented as formula (Id), each Nb independently represents an oligonucleotide sequence comprising 0-10, 0-7, 0-5, 0-4, 0-2, or 0 modified nucleotides. Preferably, Nb is 0, 1, 2, 3, 4, 5, or 6. Each Na can independently represent an oligonucleotide sequence comprising 2-20, 215, or 2-10 modified nucleotides.

[00267] Each X, Y, and Z can be the same or different from each other.

[00268] In other modes, i is 0 and j is 0, and the sense tape can be represented by the formula: 5' np-Na-YYY-na-nq 3' (Ia).

[00269] When the sense strand is represented by formula (Ia), each Na can independently represent an oligonucleotide sequence comprising 2-20, 2-15 or 2-10 modified nucleotides.

[00270] In one embodiment, the RNAi antisense strand sequence can be represented by formula (II): 5' nq'-Na'-(Z'Z'Z')k-Nb'-Y'Y'Y'-Nb'-(X'X'X')l-N'a-np' 3' (II) where: Petition 870260042207, dated 05 / 05 / 2026, page 108 / 431 / 386 kel are, each one, independently 0 or 1; p' and q' are each independently 0-6; Each Na' independently represents an oligonucleotide sequence comprising 0-25 modified nucleotides, each sequence comprising at least two differently modified nucleotides; Each Nb' independently represents an oligonucleotide sequence comprising 0-10 modified nucleotides; Each np' and nq' independently represent a salient nucleotide; where Nb' and Y' do not have the same modification; and X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent a motif of three identical modifications in three consecutive nucleotides.

[00271] In one embodiment, Na' and / or Nb' comprise alternating pattern modifications.

[00272] The Y'Y'Y' motif occurs near or at the cleavage site of the antisense strand. For example, when the RNAi agent has a duplex region of 1723 nucleotides in length, the Y'Y'Y' motif may occur at positions 9, 10, 11; 10, 11, 12; 11, 12, 13; 12, 13, 14; or 13, 14, 15 of the antisense strand, with counting starting from the 1st nucleotide, from the 5' end; or optionally, counting starting at the 1st paired nucleotide within the duplex region, from the 5' end. Preferably, the Y'Y'Y' motif occurs at positions 11, 12, 13.

[00273] In one embodiment, the Y'Y'Y' motif is all nucleotides modified with 2'-OMe.

[00274] In one embodiment, k is 1 and el is 0, or k is 0 and el is 1, or kel are both 1.

[00275] The antisense tape can therefore be represented by Petition 870260042207, dated 05 / 05 / 2026, page 109 / 431 / 386 following formulas: 5' nq'-Na'-Z'Z'Z'-Nb'-Y'Y'Y'-Na'-np' 3' (IIb); 5' nq'-Na'-Y'Y'Y'-Nb'-X'X'X'-np' 3' (IIc); or 5' nq'-Na'- Z'Z'Z'-Nb'-Y'Y'Y'-Nb'-X'X'X'-Na'-np' 3' (IId).

[00276] When the antisense strand is represented by formula (IIb), Nb' represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2 or 0 modified nucleotides. Each Na' independently represents an oligonucleotide sequence comprising 2-20, 215 or 2-10 modified nucleotides.

[00277] When the antisense strand is represented as formula (IIc), Nb' represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2 or 0 modified nucleotides. Each Na' independently represents an oligonucleotide sequence comprising 2-20, 215 or 2-10 modified nucleotides.

[00278] When the antisense strand is represented as formula (IId), each Nb' independently represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2, or 0 modified nucleotides. Each Na' independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. Preferably, Nb is 0, 1, 2, 3, 4, 5, or 6.

[00279] In other modes, k is 0 and el is 0, and the antisense strip can be represented by the formula: 5' np'-Na'-Y'Y'Y'-na'-nq' 3' (Ia).

[00280] When the antisense strand is represented as formula (IIa), each Na' independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides.

[00281] Each X', Y', and Z' can be the same or different from each other.

[00282] Each nucleotide of the sense strand and the antisense strand can be Petition 870260042207, dated 05 / 05 / 2026, p. 110 / 431 / 386 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 the sense and antisense strands is independently modified with 2'-O-methyl or 2'-fluoro. Each X, Y, Z, X', Y' and Z', in particular, may represent a 2'-O-methyl modification or a 2'-fluoro modification.

[00283] In one embodiment, the RNAi agent sense strand may contain the YYY motif at positions 9, 10, and 11 of the strand when the duplex region is 21 nt long, counting from the 1st nucleotide from the 5' end, or optionally, counting from the 1st paired nucleotide within the duplex region, from the 5' end; and Y represents a 2'-F modification. The sense strand may additionally contain an XXX motif or ZZZ motifs as wing-shaped modifications at the opposite end of the duplex region; and each XXX and ZZZ independently represents a 2'-OMe modification or a 2'-F modification.

[00284] In one embodiment, the antisense strand may contain the Y'Y'Y' motif at positions 11, 12, 13 of the strand, counting from the 1st nucleotide from the 5' end, or optionally, counting from the 1st paired nucleotide within the duplex region, from the 5' end; and Y' represents a 2'-O-methyl modification. The antisense strand may further contain an X'X'X' motif or Z'Z'Z' motifs as wing-shaped modifications at the opposite end of the duplex region; and each X'X'X' and Z'Z'Z' independently represents a 2'-OMe modification or a 2'-F modification.

[00285] The sense tape represented by any of the formulas (Ia), (Ib), (Ic) and (Id) above forms a duplex with an antisense tape being represented by any of the formulas (IIa), (IIb), (IIc) and (IId), respectively.

[00286] Consequently, RNAi agents for use in the methods of Petition 870260042207, dated 05 / 05 / 2026, page 111 / 431 / 386 invention may comprise a sense strand and an antisense strand, each strand having 14 to 30 nucleotides, the RNAi duplex represented by formula (III): sense: 5' np-na-(XXX)i-nb-YYY-Nb -(ZZZ)j-Na-nq 3' antisense: 3' np'-Na'-(X'X'X')k-Nb'-Y'Y'Y'-Nb'-(Z'Z'Z')lNa'-nq' 5' (III) where: i, j, kel are each independently 0 or 1; p, p', qe q' are, each independently, 0-6; Each Na and Na' independently represents an oligonucleotide sequence comprising 0-25 modified nucleotides, each sequence comprising at least two differently modified nucleotides; Each Nb and Nb' independently represents an oligonucleotide sequence comprising 0-10 modified nucleotides; where each np', np, nq' and nq, which may or may not each be present, independently represents a salient nucleotide; and XXX, YYY, ZZZ, X'X'X', Y'Y'Y', and Z'Z'Z' each independently represent a motif of three identical modifications in three consecutive nucleotides.

[00287] 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 el is 0; or k is 1 and el is 0; k is 0 and el is 1; or kel are both 0; or kel are both 1.

[00288] Exemplary combinations of the sense strand and the antisense strand that form an RNAi duplex include the formulas below: Petition 870260042207, dated 05 / 05 / 2026, p. 112 / 431 / 386

[00289] When the RNAi agent is represented by the formula (IlIa), each Na independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides.

[00290] When the RNAi agent is represented by the formula (IIIb), each Nb independently represents an oligonucleotide sequence comprising 1-10, 1-7, 1-5, or 1-4 modified nucleotides. Each Na independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides.

[00291] When the RNAi agent is represented as formula (IIIc), each Nb, Nb' independently represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2 or 0 modified nucleotides. Each Na independently represents an oligonucleotide sequence comprising 2-20, 2-15 or 2-10 modified nucleotides.

[00292] When the RNAi agent is represented as formula (IIId), each Nb, Nb' independently represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2, or 0 modified nucleotides. Each Na, Na' independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. Each Na, Na', Nb, and Nb' independently comprises Petition 870260042207, dated 05 / 05 / 2026, page 113 / 431 / 386 alternating pattern modifications.

[00293] Each X, Y and Z in formulas (III), (IlIa), (IlIb), (IIIc) and (IlId) can be the same or different from each other.

[00294] When the RNAi agent is represented by the formula (III), (IIIa), (IIIb), (IIIc) and (IIId), at least one of the Y nucleotides can form a base pair with one of the Y' nucleotides. Alternatively, at least two of the Y nucleotides form base pairs with the corresponding Y' nucleotides; or all three Y nucleotides form base pairs with the corresponding Y' nucleotides.

[00295] When the RNAi agent is represented by the formula (IIIb) or (IIId), at least one of the Z nucleotides can form a base pair with one of the Z' nucleotides. Alternatively, at least two of the Z nucleotides form base pairs with the corresponding Z' nucleotides; or all three Z nucleotides form base pairs with the corresponding Z' nucleotides.

[00296] When the RNAi agent is represented as formula (IIIc) or (IIId), at least one of the X nucleotides can form a base pair with one of the X' nucleotides. Alternatively, at least two of the X nucleotides form base pairs with the corresponding X' nucleotides; or all three X nucleotides form base pairs with the corresponding X' nucleotides.

[00297] In one embodiment, the modification at nucleotide Y is different from the modification at nucleotide Y', the modification at nucleotide Z is different from the modification at nucleotide Z', and / or the modification at nucleotide X is different from the modification at nucleotide X'.

[00298] When the RNAi agent is represented by formula (IIIa), each Na independently represents an oligonucleotide sequence comprising 2-20, 2-15 or 2-10 modified nucleotides.

[00299] In one embodiment, when the RNAi agent is represented Petition 870260042207, dated 05 / 05 / 2026, page 114 / 431 / 386, by formula (IIId), the Na modifications are 2'-O-methyl or 2'-fluoro modifications. In another embodiment, when the RNAi agent is represented by formula (IIId), the Na modifications are 2'-O-methyl or 2'-fluoro modifications and np'>0 and at least one np' is linked to a neighboring nucleotide through a phosphorothioate linkage. In yet another embodiment, when the RNAi agent is represented by formula (IIId), the Na modifications are 2'-O-methyl or 2'-fluoro modifications, np'>0 and at least one np' is linked to a neighboring nucleotide via a phosphorothioate linkage and the sense strand is conjugated to one or more GalNAc derivatives attached via a bivalent or trivalent branched linker (described below).In another embodiment, when the RNAi agent is represented by the formula (IIId), the Na modifications are 2'-O-methyl or 2'-fluoro modifications, np'>0 and at least one np' is linked to a neighboring nucleotide via a phosphorothioate linkage and the sense strand is conjugated to one or more GalNAc derivatives attached via a bivalent or trivalent branched linker.

[00300] In one embodiment, when the RNAi agent is represented by formula (IIIa), the Na modifications are 2'-O-methyl or 2'-fluoro modifications, np'>0 and at least one np' is linked to a neighboring nucleotide via 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 via a bivalent or trivalent branched linker.

[00301] In one embodiment, the RNAi agent is a multimer containing at least two duplexes represented by the formulas (III0, (IIIa), (IIIb), (IIIc) and (IIId), wherein the duplexes are connected by a linker. The linker may be cleavable or non-cleavable. Optionally, the multimer further comprises a linker. Each of the duplexes may target the same gene or two different genes; or each of the duplexes may target the same gene at two different target sites. Petition 870260042207, dated 05 / 05 / 2026, p. 115 / 431 / 386

[00302] In one embodiment, the RNAi agent is a multimer containing three, four, five, six or more duplexes represented by the formulas (III0, (IlIa), (IlIb), (IIIc) and (IlId), wherein the duplexes are connected by a linker. The linker may be cleavable or non-cleavable. Optionally, the multimer further comprises a linker. Each of the duplexes may target the same gene or two different genes; or each of the duplexes may target the same gene at two different target sites.

[00303] In one embodiment, two RNAi agents, represented by the formulas (III0. (IIIa), (IIIb), (IIIc) and (IIId), are linked to each other at the 5' end, and one or both ends are optionally conjugated to a ligand. Each of the agents may target the same gene or two different genes; or each duplex may target the same gene at two different target sites.

[00304] In certain embodiments, an RNAi agent of the invention may contain a low number of nucleotides containing a 2'-fluoro modification, e.g., 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 one specific embodiment, the RNAi agent of the invention contains 10 nucleotides with a 2'-fluoro modification, e.g., 4 nucleotides with a 2'-fluoro modification on the sense strand and 6 nucleotides with a 2'-fluoro modification on the antisense strand. In another specific embodiment, the RNAi agent of the invention contains 6 nucleotides with a 2'-fluoro modification, e.g. For example, 4 nucleotides with a 2'-fluoro modification on the sense strand and 2 nucleotides with a 2'-fluoro modification on the antisense strand.

[00305] In other embodiments, an RNAi agent of the invention may contain an ultra-low number of nucleotides containing a 2'-fluoro modification, e.g., 2 or fewer nucleotides containing a 2'-fluoro modification. For example, the RNAi agent may contain 2, 1, or 0 nucleotides with a Petition 870260042207, dated 05 / 05 / 2026, page 116 / 431 100 / 386 2'-fluoro modification. In a specific embodiment, the RNAi agent may contain 2 nucleotides with a 2'-fluoro modification, e.g., 0 nucleotides with a 2'-fluoro modification on the sense strand and 2 nucleotides with a 2'-fluoro modification on the antisense strand.

[00306] Several publications describe multimeric RNAi agents that can be used in the methods of the invention. Such publications include WO2007 / 091269, US Patent No. 7858769, WO2010 / 141511, WO2007 / 117686, WO2009 / 014887 and WO2011 / 031520, the contents of which are incorporated herein by reference.

[00307] As described in more detail below, an RNAi agent containing conjugations of one or more carbohydrate moieties with an RNAi agent can optimize one or more properties of the RNAi agent. In many cases, the carbohydrate moiety will be 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 by another moiety, e.g., a non-carbohydrate carrier (preferably cyclic) to which a carbohydrate linker is attached. A ribonucleotide subunit in which the ribose sugar of the subunit has thus been replaced is referred to here as a ribose substitution-modified subunit (RRMS). A cyclic carrier can be a carbocyclic ring system, meaning all the atoms in the ring are carbon atoms, or a heterocyclic ring system, meaning one or more atoms in the ring can be a heteroatom, e.g., nitrogen, oxygen, sulfur.The cyclic carrier can be a monocyclic ring system or it can contain two or more rings, e.g., fused rings. The cyclic carrier can be a fully saturated ring system, or it can contain one or more double bonds.

[00308] The linker can be attached to the polynucleotide via a carrier. Carriers include (i) at least one “backbone binding site”, preferably two “backbone binding sites” and Petition 870260042207, dated 05 / 05 / 2026, page 117 / 431 101 / 386 (ii) at least one “tethering point”. A “main chain linkage”, in this descriptive report, refers to a functional group, e.g., a hydroxyl group or, generally, an available linkage that is suitable for incorporation of the carrier into the main chain, e.g., the phosphate, the modified phosphate, e.g., the sulfur-containing main chain of a ribonucleic acid. A “tethering point” (TAP), in some embodiments, refers to a constituent atom of the cyclic carrier ring, e.g., a carbon atom or a heteroatom (distinct from an atom that provides a main chain linkage), that connects a selected moiety. The moiety may be, e.g., a carbohydrate, e.g., a monosaccharide, disaccharide, trisaccharide, tetrasaccharide, oligosaccharide, and polysaccharide. Optionally, the selected portion is connected by a tether to the cyclic carrier.Thus, the cyclic carrier will often include a functional group, e.g., an amino group, or will generally provide a linkage that is suitable for incorporation or tethering of another chemical entity, e.g., a ligand to the constituent ring.

[00309] RNAi agents can be conjugated to a ligand via a carrier, wherein the carrier can be a cyclic group or an acyclic group; preferably, the cyclic group is selected from pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolane, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuryl and decalin; preferably, the acyclic group is selected from serinol backbone and diethanolamine backbone.

[00310] In another embodiment of the invention, an RNAi agent comprises a sense strand and an antisense strand, each with 14 to 40 nucleotides. The RNAi agent can be represented by the formula (L): Petition 870260042207, dated 05 / 05 / 2026, p. 118 / 431 102 / 386

[00311] In formula (L), B1, B2, B3, B1', B2', B3' and B4' are each independently a nucleotide containing a selected group modification 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 2'-OMe modifications. In certain embodiments, at least one of B1, B2, B3, B1', B2', B3' and B4' contains a 2'-ON-ethylacetamido (2'-O-NMA) modification.

[00312] C1 is a thermally destabilizing nucleotide, located opposite the seed region of the antisense strand (i.e., at positions 2-8 of the 5' end of the antisense strand). For example, C1 is in a position on the sense strand that pairs with a nucleotide at positions 2-8 of the 5' end of the sense strand. The C1 nucleotide carries the thermally destabilizing modification which may 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., non-blocking nucleic acids (UMA) or glycerol-containing nucleic acid (GNA). In certain embodiments, C1 possesses a thermally destabilizing modification selected from the group consisting of: 1) a mismatch with the opposite nucleotide on the antisense strand; ii) an abasic modification selected from the group consisting of: Petition 870260042207, dated 05 / 05 / 2026, page 119 / 431 103 / 386 and iii) sugar modification selected from the group consisting of: 2'-deoxy, wherein B is a modified or unmodified nucleobase, R1 and R2 are independently H, halogen, OR3, or alkyl; and R3 is H, alkyl, cycloalkyl, aryl, arylalkyl, heteroaryl, or sugar. In certain embodiments, the thermally destabilizing modification at C1 is a pairing error 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; and optionally, at least one nucleobase in the pairing error is a 2'-deoxy nucleobase. In one example, the thermally destabilizing modification at C1 is GNA or

[00313] T1, T1', T2' and T3' each independently represent a nucleotide comprising a modification that provides the nucleotide with a steric volume that is equal to or less than the steric volume of a 2'-OMe modification. Steric volume refers to the sum of steric effects of Petition 870260042207, dated 05 / 05 / 2026, page 120 / 431 104 / 386 a modification. Methods for determining the steric effects of a nucleotide modification are known to those skilled in the art. The modification may be at the 2' position of a ribose sugar of the nucleotide, or a modification to a non-ribose nucleotide, an acyclic nucleotide, or the nucleotide backbone that is similar or equivalent to the 2' position of the ribose sugar, and provides the nucleotide with a steric volume that is less than or equal to the steric volume 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.

[00314] n1, n3 and q1 independently have lengths of 4 to 15 nucleotides.

[00315] n5, q3 and q7 independently have 1-6 nucleotides in length.

[00316] n4, q2 and q6 independently have 1-3 nucleotides in length; alternatively, n4 is 0.

[00317] q5 has a length of 0-10 nucleotides independently.

[00318] n2e q4 independently have 0-3 nucleotides in length.

[00319] Alternatively, n4tem is 0-3 nucleotides long.

[00320] In certain embodiments, n4 can be 0. In one example, n4 is 0, and q2 and q6 are 1. In another example, n4 is 0, and q2 and q6 are 1, with two phosphorothioate linkage modifications between nucleotides at positions 1-5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate linkage modifications between nucleotides at positions 1 and 2 and two phosphorothioate linkage modifications between nucleotides at positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). Petition 870260042207, dated 05 / 05 / 2026, pp. 121 / 431 105 / 386

[00321] In certain modalities, n4, q2 and q6 are 1.

[00322] In certain modalities, n2, n4, q2, q4 and q6 are 1.

[00323] In certain embodiments, C1 is at position 14-17 of the 5' end of the sense strand, when the sense strand is 19-22 nucleotides long, and n4 is 1. In certain embodiments, C1 is at position 15 of the 5' end of the sense strand.

[00324] In certain embodiments, T3' starts at position 2 from the 5' end of the antisense tape. In one example, T3' is at position 2 from the 5' end of the antisense tape and q6 is equal to 1.

[00325] In certain embodiments, T1' starts at position 14 from the 5' end of the antisense tape. In one example, T1' is at position 14 from the 5' end of the antisense tape and q2 is equal to 1.

[00326] In an exemplary embodiment, T3' starts from position 2 from the 5' end of the antisense tape and T1' starts from position 14 from the 5' end of the antisense tape. In an example, T3' starts from position 2 from the 5' end of the antisense tape and q6 is equal to 1, and T1' starts from position 14 from the 5' end of the antisense tape and q2 is equal to 1.

[00327] In certain embodiments, T1' and T3' are separated by 11 nucleotides in length (i.e., not counting the nucleotides of T1' and T3').

[00328] 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 q2 is equal to 1, and the modification at position 2' or at positions in a non-ribose, acyclic or non-ribose backbone that provides less steric bulk than a 2'-OMe ribose.

[00329] In certain embodiments, T3' is in position 2 from the 5' end of the antisense tape. In one example, T3' is in position 2 from the 5' end of the antisense tape and q6 is equal to 1, and the modification in Petition 870260042207, dated 05 / 05 / 2026, p. 122 / 431 106 / 386 2' position or in positions in a non-ribose, acyclic backbone or one that provides steric volume less than or equal to that of a 2'-OMe ribose.

[00330] In certain embodiments, T1 is at the cleavage site of the sense strand. In one example, T1 is at position 11 from the 5' end of the sense strand, when the sense strand is 19-22 nucleotides long and n2 is 1. In an exemplary embodiment, T1 is at the cleavage site of the sense strand at position 11 from the 5' end of the sense strand, when the sense strand is 19-22 nucleotides long and n2 is 1. In certain configurations, T2' starts at position 6 from the 5' end of the antisense tape. In one example, T2' is at positions 6-10 from the 5' end of the antisense tape and q4 is 1.

[00331] In an exemplary embodiment, T1 is at the phytasense cleavage site, for example, at position 11 from the 5' end of the sense strand, when the sense strand is 19-22 nucleotides long and n2 is 1; T1' is at position 14 from the 5' end of the antisense strand, and q2 is equal to 1, and the modification to T1' is at position 2' of a ribose sugar or at positions in a non-ribose, acyclic backbone or one that provides less steric bulk than a 2'-OMe ribose; T2' is at positions 6-10 from the 5' end of the antisense strand and q4 is 1; and T3' is at position 2 from the 5' end of the antisense strand and q6 is equal to 1, and the modification to T3' is at position 2' or at positions in a non-ribose, acyclic backbone or one that provides steric bulk less than or equal to that of a 2'-OMe ribose.

[00332] In certain embodiments, T2' starts at position 8 from the 5' end of the antisense tape. In one example, T2' starts at position 8 from the 5' end of the antisense tape and q4 is 2.

[00333] In certain embodiments, T2' starts at position 9 from the 5' end of the antisense tape. In one example, T2' is at position 9 from the 5' end of the antisense tape and q4 is 1.

[00334] In certain modalities, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is Petition 870260042207, dated 05 / 05 / 2026, page 123 / 431 107 / 386 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 1, B3' is 2'-OMe or 2'-F, q5 is 6, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate linkage modifications between nucleotides at positions 1 and 2 and two phosphorothioate linkage modifications between nucleotides at positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[00335] In certain modalities, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'-F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 1, B3' is 2'-OMe or 2'-F, q5 is 6, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate linkage modifications between nucleotides at positions 1 and 2 and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[00336] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1.

[00337] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate linkage modifications between nucleotides at positions 1 and 2 and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the sense strand). Petition 870260042207, dated 05 / 05 / 2026, pp. 124 / 431 108 / 386 antisense tape).

[00338] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 6, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 7, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1.

[00339] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 6, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 7, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate linkage modifications between nucleotides at positions 1 and 2 and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[00340] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 1, B3' is 2'-OMe or 2'-F, q5 is 6, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1.

[00341] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 1, B3' is 2'-OMe or 2'-F, q5 is 6, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate linkage modifications between nucleotides at positions 1 and 2 and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). Petition 870260042207, dated 05 / 05 / 2026, page 125 / 431 109 / 386

[00342] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 5, T2' is 2'-F, q4 is 1, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; optionally with at least 2 additional TTs at the 3' end of the antisense tape.

[00343] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 5, T2' is 2'-F, q4 is 1, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; optionally with at least 2 additional TTs at the 3' end of the antisense tape; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate linkage modifications between nucleotides at positions 1 and 2 and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[00344] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1.

[00345] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end), and two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within the positions Petition 870260042207, dated 05 / 05 / 2026, page 126 / 431 110 / 386 18-23 of the antisense tape (counting from the 5' end).

[00346] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1.

[00347] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate linkage modifications between nucleotides at positions 1 and 2 and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand).

[00348] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1.

[00349] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), and two phosphorothioate linkage modifications between nucleotides at positions 1 and 2 and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). Petition 870260042207, dated 05 / 05 / 2026, page 127 / 431 111 / 386

[00350] The RNAi agent may comprise a phosphorus-containing group at the 5' end of the sense strand or the antisense strand. The phosphorus-containing group at the 5' end may be 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 ( ). When the phosphorus-containing group at the 5' end is 5'-vinylphosphonate (5'-VP), the 5'-VP may be the 5'-E-VP isomer (i.e., trans-vinylphosphate, ), the 5'-Z-VP isomer (i.e., cis-vinylphosphate, ), or mixtures thereof.

[00351] 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.

[00352] In certain embodiments, the RNAi agent comprises a 5'-P. In certain embodiments, the RNAi agent comprises a 5'-P on the antisense strand.

[00353] In certain embodiments, the RNAi agent comprises a 5'-PS. In certain embodiments, the RNAi agent comprises a 5'-PS on the antisense strand.

[00354] In certain embodiments, the RNAi agent comprises a 5'VP. In certain embodiments, the RNAi agent comprises a 5'-VP on the antisense strand. In certain embodiments, the RNAi agent comprises a 5'-E-VP on the antisense strand. In certain embodiments, the RNAi agent comprises a 5'Z-VP on the antisense strand.

[00355] In certain modalities, the RNAi agent comprises a 5' Petition 870260042207, dated 05 / 05 / 2026, page 128 / 431 112 / 386 PS2. In certain embodiments, the RNAi agent comprises a 5'-PS2 on the antisense tape.

[00356] In certain embodiments, the RNAi agent comprises a 5'PS2. In certain embodiments, the RNAi agent comprises a 5'-deoxy-5'-Cmalonyl in the antisense strand.

[00357] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1. The RNAi agent also comprises 5'-PS.

[00358] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1. The RNAi agent also comprises 5'-P.

[00359] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1. The RNAi agent also comprises 5'-VP. 5'-VP can be 5'-E-VP, 5'-ZVP, or a combination thereof.

[00360] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1. The RNAi agent also comprises 5'-PS2.

[00361] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, Petition 870260042207, dated 05 / 05 / 2026, page 129 / 431 113 / 386 B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1. The RNAi agent also comprises 5'-deoxy-5'-C-malonyl.

[00362] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-P.

[00363] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-PS.

[00364] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 Petition 870260042207, dated 05 / 05 / 2026, pp. 130 / 431 114 / 386 and 2 and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-VP. The 5'-VP can be 5'E-VP, 5'-Z-VP, or a combination thereof.

[00365] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-PS2.

[00366] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-deoxy-5'-C-malonyl.

[00367] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1. The RNAi agent also comprises 5'-P. Petition 870260042207, dated 05 / 05 / 2026, pages 131 / 431 115 / 386

[00368] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1. The dsRNA agent also comprises 5'-PS.

[00369] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1. The RNAi agent also comprises 5'-VP. 5'-VP can be 5'-E-VP, 5'-Z-VP, or a combination thereof.

[00370] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1. The RNAi agent also comprises 5'-PS2.

[00371] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1. The RNAi agent also comprises 5'-deoxy-5'-C-malonyl.

[00372] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within the positions Petition 870260042207, dated 05 / 05 / 2026, page 132 / 431 116 / 386 18-23 of the antisense strand (counting from the 5' end). The RNAi agent also comprises 5'-P.

[00373] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end). The RNAi agent also comprises 5'-PS.

[00374] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end). The RNAi agent also comprises 5'-VP. The 5'-VP can be 5'-E-VP, 5'-Z-VP, or a combination thereof.

[00375] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end), two modifications of Petition 870260042207, dated 05 / 05 / 2026, page 133 / 431 117 / 386 phosphorothioate linkages between nucleotides at positions 1 and 2 and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end). The RNAi agent also comprises 5'-PS2.

[00376] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end). The RNAi agent also comprises 5'-deoxy-5'-C-malonyl.

[00377] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1. The RNAi agent also comprises 5'-P.

[00378] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1. The RNAi agent also comprises 5'-PS.

[00379] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1. The RNAi agent also comprises 5'-VP. The 5'-VP can be 5'-E-VP, 5'-Z-VP or Petition 870260042207, dated 05 / 05 / 2026, page 134 / 431 118 / 386 a combination of the same.

[00380] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1. The RNA dsRNA agent also comprises 5'-PS2.

[00381] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1. The RNAi agent also comprises 5'-deoxy-5'-C-malonyl.

[00382] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-P.

[00383] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within the Petition 870260042207, dated 05 / 05 / 2026, page 135 / 431 119 / 386 positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-PS.

[00384] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-VP. The 5'-VP can be 5'E-VP, 5'-Z-VP, or a combination thereof.

[00385] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-PS2.

[00386] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two Petition 870260042207, dated 05 / 05 / 2026, page 136 / 431 120 / 386 phosphorothioate linkage modifications between nucleotides at positions 1 and 2 and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-deoxy-5'-C-malonyl.

[00387] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1. The RNAi agent also comprises 5'-P.

[00388] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1. The RNAi agent also comprises 5'-PS.

[00389] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1. The RNAi agent also comprises 5'-VP. 5'-VP can be 5'-E-VP, 5'-Z-VP, or a combination thereof.

[00390] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1. The RNAi agent also comprises 5'-PS2.

[00391] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1. The RNAi agent Petition 870260042207, dated 05 / 05 / 2026, page 137 / 431 121 / 386 also includes 5'-deoxy-5'-C-malonyl.

[00392] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-P.

[00393] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-PS.

[00394] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within the Petition 870260042207, dated 05 / 05 / 2026, page 138 / 431 122 / 386 positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-VP. The 5'-VP can be 5'E-VP, 5'-Z-VP, or a combination thereof.

[00395] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-PS2.

[00396] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-deoxy-5'-C-malonyl.

[00397] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), Petition 870260042207, dated 05 / 05 / 2026, page 139 / 431 123 / 386 two phosphorothioate linkage modifications between nucleotides at positions 1 and 2 and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 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.

[00398] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 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.

[00399] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-VP (e.g., 5'-E-VP, 5' Petition 870260042207, dated 05 / 05 / 2026, pp. 140 / 431 124 / 386 Z-VP (or a combination thereof), and a targeting ligand.

[00400] In certain embodiments, the 5'-VP is at the 5' end of the antisense tape, and the targeting link is at the 3' end of the sense tape.

[00401] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-PS2e and a targeting ligand.In certain configurations, the 5'-PS2 connector is located at the 5' end of the antisense tape, and the targeting connector is located at the 3' end of the sense tape.

[00402] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 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. Petition 870260042207, dated 05 / 05 / 2026, pp. 141 / 431 125 / 386

[00403] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end). The RNAi agent also comprises 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.

[00404] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end). The RNAi agent also comprises 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.

[00405] In certain embodiments, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within the positions Petition 870260042207, dated 05 / 05 / 2026, page 142 / 431 126 / 386 RNAi consists of two phosphorothioate linkage modifications between nucleotides at positions 1-5 of the sense strand (counting from the 5' end), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end). The RNAi agent also comprises 5'-VP (e.g., 5'-E-VP, 5'-Z-VP, or a combination thereof) and a targeting linker. In certain embodiments, the 5'VP is at the 5' end of the antisense strand, and the targeting linker is at the 3' end of the sense strand.

[00406] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end). The RNAi agent also comprises 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.

[00407] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-OMe and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end). The RNAi agent Petition 870260042207, dated 05 / 05 / 2026, pages 143 / 431 127 / 386 also comprises 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.

[00408] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 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.

[00409] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 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. Petition 870260042207, dated 05 / 05 / 2026, pp. 144 / 431 128 / 386

[00410] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 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.

[00411] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 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.

[00412] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, T2' is 2'-F, q4 is 2, Petition 870260042207, dated 05 / 05 / 2026, page 145 / 431 129 / 386 B3' is 2'-OMe or 2'-F, q5 is 5, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 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.

[00413] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 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.

[00414] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2 and. Petition 870260042207, dated 05 / 05 / 2026, page 146 / 431 130 / 386 two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-PS and a targeting linker. In certain embodiments, the 5'-PS is at the 5' end of the antisense strand, and the targeting linker is at the 3' end of the sense strand.

[00415] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 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.

[00416] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 5'-PS2e and a ligand of Petition 870260042207, dated 05 / 05 / 2026, page 147 / 431 131 / 386 routing. In certain configurations, the 5'-PS2 is at the 5' end of the antisense tape, and the routing linker is at the 3' end of the sense tape.

[00417] In certain modalities, B1 is 2'-OMe or 2'-F, n1 is 8, T1 is 2'F, n2 is 3, B2 is 2'-OMe, n3 is 7, n4 is 0, B3 is 2'-OMe, n5 is 3, B1' is 2'-OMe or 2'F, q1 is 9, T1' is 2'-F, q2 is 1, B2' is 2'-OMe or 2'-F, q3 is 4, q4 is 0, B3' is 2'-OMe or 2'-F, q5 is 7, T3' is 2'-F, q6 is 1, B4' is 2'-F and q7 is 1; with two phosphorothioate linkage modifications between nucleotides within positions 1-5 of the sense strand (counting from the 5' end of the sense strand), two phosphorothioate linkage modifications between nucleotides at positions 1 and 2, and two phosphorothioate linkage modifications between nucleotides within positions 18-23 of the antisense strand (counting from the 5' end of the antisense strand). The RNAi agent also comprises 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.

[00418] In one particular embodiment, an RNAi agent of the present invention comprises: (a) a sense tape having: (i) a length of 21 nucleotides; (ii) an ASGPR linker attached to the 3' end, wherein said ASGPR linker comprises three GalNAc derivatives attached via a trivalent branched linker; and (iii) 2'-F modifications at positions 1, 3, 5, 7, 9 to 11, 13, 17, 19 and 21, and 2'-OMe modifications at positions 2, 4, 6, 8, 12, 14 to 16, 18 and 20 (counting from the 5' end); and (b) an antisense tape having: (i) a length of 23 nucleotides; Petition 870260042207, dated 05 / 05 / 2026, pp. 148 / 431 132 / 386 (ii) 2'-OMe modifications at positions 1, 3, 5, 9, 11 to 13, 15, 17, 19, 21 and 23, 2'F modifications at positions 2, 4, 6 to 8, 10, 14, 16, 18, 20 and 22 (counting from the 5' end); and (iii) phosphorothioate linkages between the nucleotides at positions 21 and 22 and between the nucleotides at positions 22 and 23 (counting from the 5' end); whereby the dsRNA agents have a two-nucleotide overhang at the 3' end of the antisense strand, and a blunt end at the 5' end of the antisense strand.

[00419] In another particular embodiment, an RNAi agent of the present invention comprises: (a) a sense tape having: (i) a length of 21 nucleotides; (ii) an ASGPR linker attached to the 3' end, wherein said ASGPR linker comprises three GalNAc derivatives attached via a trivalent branched linker; (iii) 2'-F modifications at positions 1, 3, 5, 7, 9 to 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 linkages between nucleotides at positions 1 and 2 and between nucleotides at positions 2 and 3 (counting from the 5' end); and (b) an antisense tape having: (i) a length of 23 nucleotides; (ii) 2'-OMe modifications at positions 1, 3, 5, 7, 9, 11 to 13, 15, 17, 19 and 21 to 23, 2'F modifications at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20 (counting from the 5' end); and (iii) phosphorothioate linkages between the nucleotides at positions 1 Petition 870260042207, dated 05 / 05 / 2026, page 149 / 431 133 / 386 and 2, between the nucleotides at positions 2 and 3, between the nucleotides at positions 21 and 22, and between the nucleotides at positions 22 and 23 (counting from the 5' end); in which RNAi agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00420] In another particular embodiment, an RNAi agent of the present invention comprises: (a) a sense tape having: (i) a length of 21 nucleotides; (ii) an ASGPR linker attached to the 3' end, wherein said ASGPR linker comprises three GalNAc derivatives attached via 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 linkages between the nucleotides at positions 1 and 2 and between the nucleotides at positions 2 and 3 (counting from the 5' end); and (b) an antisense tape having: (i) a length of 23 nucleotides; (ii) 2'-OMe modifications at positions 1, 3, 7, 9, 11, 13, 15, 17 and 19 to 23 and 2'-F modifications at positions 2, 4 to 6, 8, 10, 12, 14, 16 and 18 (counting from the 5' end); and (iii) phosphorothioate linkages between nucleotides at positions 1 and 2, between nucleotides at positions 2 and 3, between nucleotides at positions 21 and 22 and between nucleotides at positions 22 and 23 (counting from the 5' end); Petition 870260042207, dated 05 / 05 / 2026, page 150 / 431 134 / 386 in which RNAi agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00421] In another particular embodiment, an RNAi agent of the present invention comprises: (a) a sense tape having: (i) a length of 21 nucleotides; (ii) an ASGPR linker attached to the 3' end, wherein said ASGPR linker comprises three GalNAc derivatives attached via 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 linkages between nucleotides at positions 1 and 2 and between nucleotides at positions 2 and 3 (counting from the 5' end); and (b) an antisense tape having: (i) a length of 23 nucleotides; (ii) 2'-OMe modifications at positions 1, 5, 7, 9, 11, 13, 15, 17, 19 and 21 to 23, and 2'-F modifications at positions 2 to 4, 6, 8, 10, 12, 14, 16, 18 and 20 (counting from the 5' end); and (iii) phosphorothioate linkages between nucleotides at positions 1 and 2, between nucleotides at positions 2 and 3, between nucleotides at positions 21 and 22 and between nucleotides at positions 22 and 23 (counting from the 5' end); in which RNAi agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00422] In another particular embodiment, an RNAi agent of Petition 870260042207, dated 05 / 05 / 2026, page 151 / 431 135 / 386 The present invention comprises: (a) a sense tape having: (i) a length of 21 nucleotides; (ii) an ASGPR linker attached to the 3' end, wherein said ASGPR linker comprises three GalNAc derivatives attached via 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 linkages between nucleotides at positions 1 and 2 and between nucleotides at positions 2 and 3 (counting from the 5' end); and (b) an antisense tape having: (i) a length of 23 nucleotides; (ii) 2'-OMe modifications at positions 1, 3, 5, 7, 9, 11 to 13, 15, 17, 19 and 21 to 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 linkages between nucleotides at positions 1 and 2, between nucleotides at positions 2 and 3, between nucleotides at positions 21 and 22 and between nucleotides at positions 22 and 23 (counting from the 5' end); in which RNAi agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00423] In another particular embodiment, an RNAi agent of the present invention comprises: (a) a sense tape having: (i) a length of 21 nucleotides; (ii) an ASGPR linker attached to the 3' end, wherein the Petition 870260042207, dated 05 / 05 / 2026, page 152 / 431 136 / 386 said ASGPR linker comprises three GalNAc derivatives attached via a trivalent branched linker; (iii) 2'-F modifications at positions 1, 3, 5, 7, 9 to 11 and 13 and 2'-OMe modifications at positions 2, 4, 6, 8, 12 and 14 to 21; and (iv) phosphorothioate linkages between nucleotides at positions 1 and 2 and between nucleotides at positions 2 and 3 (counting from the 5' end); and (b) an antisense tape having: (i) a length of 23 nucleotides; (ii) 2'-OMe modifications at positions 1, 3, 5 to 7, 9, 11 to 13, 15, 17 to 19 and 21 to 23 and 2'-F modifications at positions 2, 4, 8, 10, 14, 16 and 20 (counting from the 5' end); and (iii) phosphorothioate linkages between nucleotides at positions 1 and 2, between nucleotides at positions 2 and 3, between nucleotides at positions 21 and 22 and between nucleotides at positions 22 and 23 (counting from the 5' end); in which RNAi agents have a two-nucleotide overhang at the 3' end of the antisense strand, and a blunt end at the 5' end of the antisense strand.

[00424] In another particular embodiment, an RNAi agent of the present invention comprises: (a) a sense tape having: (i) a length of 21 nucleotides; (ii) an ASGPR linker attached to the 3' end, wherein said ASGPR linker comprises three GalNAc derivatives attached via a trivalent branched linker; (iii) 2'-OMe modifications in positions 1, 2, 4, 6, 8, 12, 14, 15, 17 and 19 to 21 and 2'-F modifications in positions 3, 5, 7, 9 to 11, 13, 16 and 18; Petition 870260042207, dated 05 / 05 / 2026, page 153 / 431 137 / 386 and (iv) phosphorothioate linkages between nucleotides at positions 1 and 2 and between nucleotides at positions 2 and 3 (counting from the 5' end); and (b) an antisense tape having: (i) a length of 25 nucleotides; (ii) 2'-OMe modifications at positions 1, 4, 6, 7, 9, 11 to 13, 15, 17 and 19 to 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 linkages between nucleotides at positions 1 and 2, between nucleotides at positions 2 and 3, between nucleotides at positions 21 and 22 and between nucleotides at positions 22 and 23 (counting from the 5' end); in which RNAi agents have a four-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00425] In another particular embodiment, an RNAi agent of the present invention comprises: (a) a sense tape having: (i) a length of 21 nucleotides; (ii) an ASGPR linker attached to the 3' end, wherein said ASGPR linker comprises three GalNAc derivatives attached via 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 linkages between nucleotides at positions 1 and 2 and between nucleotides at positions 2 and 3 (counting from the Petition 870260042207, dated 05 / 05 / 2026, p. 154 / 431 138 / 386 end 5'); and (b) an antisense tape having: (i) a length of 23 nucleotides; (ii) 2'-OMe modifications at positions 1, 3 to 5, 7, 8, 10 to 13, 15 and 17 to 23 and 2'-F modifications at positions 2, 6, 9, 14 and 16 (counting from the 5' end); and (iii) phosphorothioate linkages between nucleotides at positions 1 and 2, between nucleotides at positions 2 and 3, between nucleotides at positions 21 and 22 and between nucleotides at positions 22 and 23 (counting from the 5' end); in which RNAi agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00426] In another particular embodiment, an RNAi agent of the present invention comprises: (a) a sense tape having: (i) a length of 21 nucleotides; (ii) an ASGPR linker attached to the 3' end, wherein said ASGPR linker comprises three GalNAc derivatives attached via 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 linkages between nucleotides at positions 1 and 2 and between nucleotides at positions 2 and 3 (counting from the 5' end); and (b) an antisense tape having: (i) a length of 23 nucleotides; Petition 870260042207, dated 05 / 05 / 2026, page 155 / 431 139 / 386 (ii) 2'-OMe modifications at positions 1, 3 to 5, 7, 10 to 13, 15 and 17 to 23 and 2'-F modifications at positions 2, 6, 8, 9, 14 and 16 (counting from the 5' end); and (iii) phosphorothioate linkages between nucleotides at positions 1 and 2, between nucleotides at positions 2 and 3, between nucleotides at positions 21 and 22, and between nucleotides at positions 22 and 23 (counting from the 5' end); in which RNAi agents have a two-nucleotide overhang at the 3' end of the antisense strand, and a blunt end at the 5' end of the antisense strand.

[00427] In another particular embodiment, an RNAi agent of the present invention comprises: (a) a sense tape having: (i) a length of 19 nucleotides; (ii) an ASGPR linker attached to the 3' end, wherein said ASGPR linker comprises three GalNAc derivatives attached via 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 linkages between nucleotides at positions 1 and 2 and between nucleotides at positions 2 and 3 (counting from the 5' end); and (b) an antisense tape having: (i) a length of 21 nucleotides; (ii) 2'-OMe modifications at positions 1, 3 to 5, 7, 10 to 13, 15 and 17 to 21 and 2'-F modifications at positions 2, 6, 8, 9, 14 and 16 (counting from the 5' end); and (iii) phosphorothioate linkages between the nucleotides at positions 1 Petition 870260042207, dated 05 / 05 / 2026, page 156 / 431 140 / 386 and 2, between the nucleotides at positions 2 and 3, between the nucleotides at positions 19 and 20, and between the nucleotides at positions 20 and 21 (counting from the 5' end); in which RNAi agents have a two-nucleotide overhang at the 3' end of the antisense strand and a blunt end at the 5' end of the antisense strand.

[00428] 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 AD413639. In one embodiment, the agent is AD-413644. In another embodiment, the agent is AD-413669. These agents may further comprise a linker. IV. iRNAs conjugated to Linkers

[00429] Another modification of the RNA of an 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, among others, lipid moieties such as a cholesterol moiety (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), a thioether, p. e.g., 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), a thiocholesterol (Oberhauser et al., (1992) Nucl. Acids Res., 20:533-538), an aliphatic chain, p. e.g., 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), a phospholipid, p. ex., dihexadecyl-rac-glycerol or triethylammonium 1,2di-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), a polyamine or a polyethylene glycol chain. Petition 870260042207, dated 05 / 05 / 2026, p. 157 / 431 141 / 386 (Manoharan et al., (1995) Nucleosides & Nucleotides, 14:969-973), or adamantane acetic acid (Manoharan et al., (1995) Tetrahedron Lett., 36:36513654), a palmityl moiety (Mishra et al., (1995) Biochim. Biophys. Acta, 1264:229-237) or an octadecylamine or hexylaminocarbonyloxycholesterol moiety (Crooke et al., (1996) J. Pharmacol. Exp. Ther., 277:923-937).

[00430] In one embodiment, a ligand alters the distribution, targeting, or lifetime of an iRNA agent in which it is incorporated. In preferred embodiments, a ligand increases the affinity for a selected target, e.g., molecule, cell or cell type, compartment, e.g., a cellular or organ compartment, tissue, organ, body region, compared, e.g., to a species without such a ligand. Preferred ligands will not be part of duplex pairing in a duplex nucleic acid.

[00431] The linkers may include a natural substance, such as a protein (e.g., human serum albumin (HSA), low-density lipoprotein (LDL), or globulin); a carbohydrate (e.g., a dextran, pullulan, chitin, chitosan, inulin, cyclodextrin, N-acetylglucosamine, N-acetylgalactosamine, or hyaluronic acid); or a lipid. The linker may also be a recombinant or synthetic molecule, such as a synthetic polymer, e.g., a polyamino acid. Examples of polyamino acids include polylysine (PLL), poly-L-aspartic acid, poly-L-glutamic acid, amino acid, styrene-maleic acid anhydride copolymer, poly(L-lactide-co-glycolide) copolymer, divinyl ether-maleic anhydride copolymer, N-(2-hydroxypropyl)methacrylamide (HMPA) copolymer, polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyurethane, poly(2-ethylacrylic acid), N-isopropylacrylamide polymers, or polyphosphazine.Examples of polyamines include: polyethyleneimine, polylysine (PLL), spermine, spermidine, polyamine, pseudopeptide-polyamine, peptidomimetic polyamine, dendrimer polyamine, arginine, amidine, protamine, lipid. Petition 870260042207, dated 05 / 05 / 2026, page 158 / 431 142 / 386 cationic, cationic porphyrin, quaternary salt of a polyamine or alpha-helical peptide.

[00432] Ligands may also include targeting groups, e.g., a targeting agent for a cell or tissue, e.g., a lectin, glycoprotein, lipid, or protein, e.g., an antibody, that binds to a specific cell type, such as a kidney cell. A targeting group may be a thyrotropin, melanotropin, lectin, glycoprotein, surfactant protein A, mucin carbohydrate, multivalent lactose, multivalent galactose, N-acetylgalactosamine, multivalent N-acetylglucosamine-mannose, multivalent fucose, glycosylated polyamino acids, multivalent galactose, transferrin, bisphosphonate, polyglutamate, polyaspartate, a lipid, cholesterol, a steroid, bile acid, folate, vitamin B12, vitamin A, biotin, or an RGD peptide or RGD peptide mimetic.

[00433] Other examples of ligands include dyes, intercalating agents (e.g., acridines), crosslinking agents (e.g., psoralen, mitomycin C), porphyrins (TPPC4, texafirin, saphrin), polycyclic aromatic hydrocarbons (e.g., phenazine, dihydrophenazine), artificial endonucleases (e.g., EDTA), lipophilic molecules, p. e.g., cholesterol, cholic acid, acetic acid, adamantane, 1-pyrenebutyric acid, dihydrotestosterone, 1,3-BisO(hexadecyl)glycerol, geranyloxyhexyl group, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl)lithocholic acid, O3-(oleoyl)cholenic acid, dimethoxytrityl or phenoxazine) and conjugated peptides (e.g., Antennapedia, Tat peptide), alkylating agents, phosphate, amino, mercapto, PEG (e.g., PEG-40K), MPEG, [MPEG]2, polyamino, alkyl, substituted alkyl, radiolabeled markers, enzymes, haptens (e.g., biotin), transport / absorption enhancers (e.g., aspirin, vitamin E, folic acid), synthetic ribonucleases (e.g., imidazole, bisimidazole, histamine, imidazole clusters,. Petition 870260042207, dated 05 / 05 / 2026, page 159 / 431 143 / 386 acridine-imidazole conjugates, Eu3+ tetra-azamacrocycle complexes), dinitrophenyl, HRP or AP.

[00434] Ligands can be proteins, e.g., glycoproteins or peptides, e.g., molecules having a specific affinity for a co-ligand, or antibodies, e.g., an antibody that binds to a specified cell type, such as a liver cell. Ligands can also include hormones and hormone receptors. They can also include non-peptidic species, such as lipids, lectins, carbohydrates, vitamins, cofactors, multivalent lactose, multivalent galactose, multivalent N-acetyl-α-N-acetylglucosamine-mannose, or multivalent fucose. The ligand can be, for example, a lipopolysaccharide, a p38 MAP kinase activator, or an NFkB activator.

[00435] The ligand is a substance, e.g., a drug, that can increase the uptake of the iRNA agent by the cell, for example, by disrupting the cell's cytoskeleton, e.g., by disrupting the cell's microtubules, microfilaments, and / or intermediate filaments. The drug may be, for example, taxon, vincristine, vinblastine, cytochalasin, nocodazole, japlaquinoline, latrunculin A, phalloidin, swinholide A, indanocin, or mioservin.

[00436] In some embodiments, a ligand attached to an iRNA, as described herein, acts as a pharmaceutical modulator (PK modulator). PK modulators include lipophiles, bile acids, steroids, phospholipid analogs, peptides, protein-binding agents, PEG, vitamins, etc. Exemplary PK modulators include, among others, cholesterol, fatty acids, cholic acid, lithocholic acid, dialkylglycerides, diacylglycerides, phospholipids, sphingolipids, naproxen, ibuprofen, vitamin E, biotin, etc. Oligonucleotides comprising multiple phosphorothioate linkages are also known to bind to serum protein; thus, short oligonucleotides, e.g., oligonucleotides with approximately 5 bases, 10 bases, 15 Petition 870260042207, dated 05 / 05 / 2026, page 160 / 431 144 / 386 bases or 20 bases, comprising multiple phosphorothioate linkages in the main chain, are also accessible for the present invention as ligands (e.g., as PK modulating ligands). Furthermore, aptamers that bind to serum components (e.g., serum proteins) are also suitable for use as PK modulating ligands in the embodiments described herein.

[00437] Ligand-conjugated oligonucleotides of the invention can be synthesized using an oligonucleotide carrying a pending reactive functionality, such as that derived from coupling a linker molecule to the oligonucleotide (described below). This reactive oligonucleotide can be reacted directly with commercially available ligands, ligands that are synthesized carrying any one of a variety of protecting groups, or ligands that have a linker moiety attached to them.

[00438] The oligonucleotides used in the conjugates of the present invention can be conveniently and routinely prepared using the well-known technique of solid-phase synthesis. Equipment for such synthesis is sold by various suppliers, including, for example, Applied Biosystems (Foster City, California). Any other means for such synthesis known in the art may be employed in addition to or alternatively. The use of similar techniques to prepare other oligonucleotides, such as phosphorothioate and alkylated derivatives, is also known.

[00439] In oligonucleotides conjugated to the ligand and ligand molecule carrying sequence-specific linked nucleosides of the present invention, the oligonucleotides and oligonucleosides can be assembled in a suitable DNA synthesizer using standard precursors of the nucleotides or nucleosides, or conjugated precursors of the nucleotides or nucleosides that already carry the linking moiety, precursors of the ligand-nucleotide or nucleoside conjugate that already carry the ligand molecule, or building blocks not carrying the nucleoside ligand. Petition 870260042207, dated 05 / 05 / 2026, page 161 / 431 145 / 386

[00440] When nucleotide conjugate precursors that already carry a linking moiety are used, the synthesis of sequence-specific linked nucleosides is typically completed, and the ligand molecule is then reacted with the linking moiety to form the ligand-conjugated oligonucleotide. In some embodiments, the oligonucleotides or linked nucleosides of the present invention are synthesized by an automated synthesizer using phosphoramidites derived from ligand-nucleoside conjugates in addition to the standard and non-standard phosphoramidites that are commercially available and routinely used in oligonucleotide synthesis. A. Conjugated Lipids

[00441] In one embodiment, the ligand or conjugate is a lipid or a lipid-based molecule. Such a lipid or lipid-based molecule preferably binds to a serum protein, e.g. (e.g., human serum albumin (HSA)). A ligand that binds to HSA allows the conjugate to be delivered to a target tissue, e.g. (e.g., a non-renal target tissue of the body). For example, the target tissue could be the liver, including hepatic parenchymal cells. Other molecules that bind to HSA may also be used as ligands. For example, neproxin or aspirin could be used. A lipid or lipid-based ligand may (a) increase the resistance to degradation of the conjugate, (b) increase targeting or transport to a target cell or cell membrane, and / or (c) be used to adjust binding to a serum protein, e.g., HSA.

[00442] A lipid-based ligand can be used to inhibit, for example, control the binding of the conjugate to a target tissue. For example, a lipid or lipid-based ligand that binds to HSA more strongly will be less likely to be targeted to the kidney and therefore less likely to be cleared from the body. A lipid or lipid-based ligand that binds to HSA less strongly can be used to target the conjugate to the kidney.

[00443] In a preferred embodiment, the lipid-based ligand binds Petition 870260042207, dated 05 / 05 / 2026, page 162 / 431 146 / 386 if to HSA. Preferably, it binds to HSA with sufficient affinity so that the conjugate is preferentially distributed to a non-renal tissue. However, it is preferred that the affinity not be so strong that the HSA-ligand binding cannot be reversed.

[00444] In another preferred embodiment, the lipid-based ligand binds to HSA weakly or not at all, such that the conjugate is preferentially distributed to the kidney. Other moieties that target renal cells may also be used in place of or in addition to the lipid-based ligand.

[00445] In another aspect, the ligand is a portion, e.g., a vitamin, that is absorbed by a target cell, e.g., a proliferating cell. These are especially useful for treating disorders characterized by unwanted cell proliferation, e.g., of the malignant or non-malignant type, e.g., cancer cells. Exemplary vitamins include vitamins A, E, and K. Other exemplary vitamins include the B vitamins, e.g., folic acid, B12, riboflavin, biotin, pyridoxal, or other vitamins or nutrients absorbed by target cells such as liver cells. HSA and low-density lipoprotein (LDL) are also included. B. Cell permeation agents

[00446] In another aspect, the ligand is a cell permeation agent, preferably a helical cell permeation agent. Preferably, the agent is amphipathic. An exemplary agent is a peptide such as Tat or Antennapedia. If it is a peptide, the agent may be modified, including with peptidylmimics, invertomers or non-peptide or pseudopeptide linkages, and the use of D-amino acids. The helical agent is preferably an alpha-helical agent, which preferably has a lipophilic and a lipophobic phase.

[00447] The ligand can be a peptide or a peptidomimetic. A peptidomimetic (also referred to here as an oligopeptidomimetic) is a Petition 870260042207, dated 05 / 05 / 2026, page 163 / 431 147 / 386 molecule capable of folding into a defined three-dimensional structure similar to that of a natural peptide. The binding of peptides and peptidomimetics to RNA agents can affect the pharmacokinetic distribution of the RNA, such as enhancing cellular recognition and uptake. The peptide or peptidomimetic portion can be approximately 5-50 amino acids in length, e.g., approximately 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 amino acids in length.

[00448] A peptide or peptidomimetic can be, for example, a cell-permeating peptide, a cationic peptide, an amphipathic peptide, or a hydrophobic peptide (e.g., consisting primarily of Tyr, Trp, or Phe). The peptide portion can be a dendrimer, hindered peptide, or cross-linked peptide. Alternatively, the peptide portion can include a hydrophobic membrane translocation sequence (MTS). An exemplary peptide containing a hydrophobic MTS is RFGF having the amino acid sequence AAVALLPAVLLALLAP (SEQ ID NO: 2977). An RFGF analog (e.g., amino acid sequence AALLPVLLAAP (SEQ ID NO:2978)) containing a hydrophobic MTS can also be a targeting moiety. The peptide moiety can be a “delivery” peptide, which can carry large polar molecules, including peptides, oligonucleotides, and protein, across cell membranes.For example, sequences of the HIV Tat protein (GRKKRRQRRRPPQ (SEQ ID NO: 2979)) and the Drosophila Antennapedia protein (RQIKIWFQNRRMKWKK (SEQ ID NO: 2980)) have been shown to function as delivery peptides. A peptide or peptidomimetic can be encoded by a random DNA sequence, such as a peptide identified in a phage expression library or a one-bead (microsphere) combinatorial compound library (OBOC) (Lam et al., Nature, 354:82-84, 1991). Examples of peptides or peptidomimetics bound to an agent. Petition 870260042207, dated 05 / 05 / 2026, page 164 / 431 148 / 386 dsRNA via an incorporated monomeric unit for cell targeting purposes is arginine-glycine-aspartic acid (RGD) peptide or RGD mimic. A peptide moiety can vary in length from approximately 5 amino acids to approximately 40 amino acids. Peptide moieties may have a structural modification, such as to increase stability or target conformational properties. Any of the structural modifications described below may be used.

[00449] An RGD peptide for use in the compositions and methods of the invention may be linear or cyclic, and may be modified, e.g., glycosylated or methylated, to facilitate targeting to specific tissue(s). Peptides and peptidomimetics containing RGD may include D-amino acids, as well as synthetic RGD mimics. In addition to RGD, other moieties targeting the integrin ligand may be used. Preferred conjugates of this ligand target PECAM-1 or VEGF.

[00450] A “cell-permeating peptide” is capable of permeating a cell, e.g., a microbial cell, such as a bacterial or fungal cell, or a mammalian cell, such as a human cell. A microbial cell-permeating peptide may be, for example, a linear α-helical peptide (e.g., LL-37 or Ceropin P1), a peptide containing a disulfide bond (e.g., α-defensin, β-defensin, or bactenecin), or a peptide containing only one or two dominant amino acids (e.g., PR-39 or indolicidin). A cell-permeating peptide may also include a nuclear localization signal (NLS). For example, a cell permeation peptide can be a bipartite amphipathic peptide, such as MPG, which is derived from the fusion peptide domain of HIV gp41 and the NLS of the large T antigen of SV40 (Simeoni et al., Nucl. Acids Res. 31:2717-2724, 2003). C. Conjugated carbohydrates Petition 870260042207, dated 05 / 05 / 2026, page 165 / 431 149 / 386

[00451] In some embodiments of the compositions and methods of the invention, an RNA oligonucleotide further comprises a carbohydrate. RNA conjugated to carbohydrates is advantageous for the in vivo delivery of nucleic acids, as well as for compositions suitable for in vivo therapeutic use, as described herein. In this descriptive report, "carbohydrate" refers to a compound that is either a carbohydrate per se formed by one or more monosaccharide units having at least 6 carbon atoms (which may be linear, branched or cyclic) with an oxygen, nitrogen or sulfur atom bonded to each carbon atom; or a compound having as part thereof a carbohydrate portion composed of one or more monosaccharide units, each having at least six carbon atoms (which may be linear, branched or cyclic), with an oxygen, nitrogen or sulfur atom bonded to each carbon atom.Representative carbohydrates include sugars (mono-, di-, tri-, and oligosaccharides containing approximately 4, 5, 6, 7, 8, or 9 monosaccharide units), and polysaccharides such as starch, glycogen, cellulose, and polysaccharide gums. Specific monosaccharides include C5 and higher sugars (e.g., C5, C6, C7, or C8); di- and trisaccharides include sugars having two or three monosaccharide units (e.g., C5, C6, C7, or C8).

[00452] In one embodiment, a conjugated carbohydrate for use in the compositions and methods of the invention is selected from the group consisting of: Petition 870260042207, dated 05 / 05 / 2026, page 166 / 431 150 / 386 H Formula III, NHAc Formula IV Formula V. HO OH ΗΟ'^Αγ. / '0HO OH NHAc HO^XY^Ov / O^ / NH Acho HOÁ nhac Formula VII, Formula VI, Petition 870260042207, dated 05 / 05 / 2026, p. 167 / 431 151 / 386 Formula IX, PO3 Formula X, O^ OH ho-XUIO HO-V-—V PO3 HO THE. HO O3P NH Formula XI, Petition 870260042207, dated 05 / 05 / 2026, p. 168 / 431 152 / 386 Formula XII, Formula XIII, Formula XIV, Formula XV, Formula XVI, Petition 870260042207, dated 05 / 05 / 2026, p. 169 / 431 153 / 386 Formula XVII, Petition 870260042207, dated 05 / 05 / 2026, p. 170 / 431 OH Mo Q© , where Y is O or S en is 3 -6 (Formula XXIV); , where Y is O or S en is 3-6 (Formula XXV); X the. OH O OH THE NHAc N \ Γ o-yOFórmula XXVI; OH HoA^On HOMMVO' NHAc HO' HO OHOO NHAc No px O-Q. HO' HO N O.PX^>O OQ OH fA ^XO-X / JN OH nhacO, where X is O or S (Formula XXVII); Petition 870260042207, dated 05 / 05 / 2026, p. 171 / 431 155 / 386 Formula XXIX; Formula XXVIII; Formula XXX; Petition 870260042207, dated 05 / 05 / 2026, p. 172 / 431 156 / 386 Formula XXXI; X Formula XXXII; Formula XXXIII. Formula XXXIV.

[00453] In another embodiment, a conjugated carbohydrate for use in the compositions and methods of the invention is a monosaccharide. In one embodiment, the monosaccharide is an N-acetylgalactosamine, such as Formula II. Petition 870260042207, dated 05 / 05 / 2026, p. 173 / 431 157 / 386

[00454] Other representative conjugated carbohydrates for use in the modalities described herein include, among others: HO HO· HO HO OH A^O AcHN HO OH -X^O AcHN HO OH A^o AcHN X / X ^0 A b0 N—rf'''^O''A''N''K''-'''~''O' 0H NO H '0 THE H H N. XOZ / A^ / Oy N The (Formula XXXVI), where one of X or Y is an oligonucleotide and the other is a hydrogen.

[00455] In certain embodiments of the invention, GalNAc or a GalNAc derivative is attached to an iRNA agent of the invention via a monovalent linker. In some embodiments, GalNAc or a GalNAc derivative is attached to an iRNA agent of the invention via a bivalent linker. In still other embodiments of the invention, GalNAc or a GalNAc derivative is attached to an iRNA agent of the invention via a trivalent linker.

[00456] In one embodiment, the double-stranded RNAi agents of the invention comprise a GalNAc or GalNAc derivative attached to the RNAi agent, e.g., at the 3' end or sense strand of a dsRNA agent as described herein. In another embodiment, the double-stranded RNAi agents of the invention comprise a plurality (e.g., 2, 3, 4, 5, or 6) of GalNAc or GalNAc derivatives, each independently attached to a plurality of nucleotides of the double-stranded RNAi agent via a plurality of monovalent linkers.

[00457] In some embodiments, for example, when the two strands of an iRNA agent of the invention are part of a molecule connected by Petition 870260042207, dated 05 / 05 / 2026, page 174 / 431 158 / 386 an unbroken chain of nucleotides between the 3' end of one strand and the 5' end of the other respective strand, forming a hairpin loop comprising a plurality of unpaired nucleotides, each unpaired nucleotide within the hairpin loop may independently comprise a GalNAc or GalNAc derivative attached via a monovalent linker.

[00458] In some embodiments, the conjugated carbohydrate also comprises one or more additional ligands as described above, such as, among others, a PK modulator and / or a cell permeation peptide.

[00459] Additional conjugated carbohydrates (and linkers) suitable for use in the present invention include those described in PCT Publications Nos. WO 2014 / 179620 and WO 2014 / 179627, the contents of which are incorporated herein in their entirety by reference. D. Linkers

[00460] In some embodiments, the conjugate or linker described herein may be attached to an iRNA oligonucleotide with multiple linkers that may be cleavable or non-cleavable.

[00461] The term “linker” or “linking group” means an organic moiety that connects two parts of a compound, e.g., covalently links two parts of a compound. Linkers typically comprise a direct link or atom, such as oxygen or sulfur, a unit such as NR8, C(O), C(O)NH, SO, SO2, SO2NH, or a chain of atoms, such as, among others, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, arylalkyl, arylalkenyl, arylalquinyl, heteroarylalkyl, heteroarylalkenyl, heteroarylquinyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalquinyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylarylalkyl, alkylarylalkenyl, alkylarylalquinyl, Petition 870260042207, dated 05 / 05 / 2026, p. 175 / 431 159 / 386 alkenylarylalkyl, alkenylarylalkyl, alkenylarylkynyl, alkynylarylalkyl, alkynylarylalkyl, alkynylarylkynyl, alkylheteroarylalkyl, alkylheteroarylalkyl, alkylheteroarylalkyl, alkenyl-heteroarylalkyl, alkenyl-heteroarylalkyl, alkenyl-heteroarylalkyl, alkenyl-heteroarylalkyl, alkynyl-heteroarylalkyl, alkynyl-heteroarylalkyl, alkynyl-heteroarylalkyl, alkylheterocyclylalkyl, alkylheterocyclylalkyl, alkylheterocyclylalkyl, alkenylheterocyclylalkyl, alkenylheterocyclylalkyl, alkenylheterocyclylalkyl, alkenyl-heterocyclylalkyl, alkynyl-heterocyclylalkyl, alkynylheterocyclylalkyl, alkynylheterocyclylalkyl, alkynyl-heterocyclylalkynyla, alkylaryl, alkenylaryl, alkynylaryl, alkylheteroaryl, alkenyl-heteroaryl, alkynylhereroaryl, we quais um or more methylenes can be interrupted or terminated by O, S, S(O), SO2, N(R8), C(O), aryl substituted or unsubstituted, heteroaryl substituted or not substituted,substituted or unsubstituted heterocyclic; where R8 is hydrogen, acyl, aliphatic or substituted aliphatic. In one embodiment, the linker has approximately 1-24 atoms, 2-24, 3-24, 4-24, 5-24, 6-24, 6-18, 7-18, 8-18 atoms, 7-17, 8-17, 6-16, 7-17 or 8-16 atoms.

[00462] A cleavable linker group is one that is sufficiently stable outside the cell, but which, upon entry into a target cell, is cleaved to release the two parts that the linker is holding together. In a preferred embodiment, the cleavable linker group is cleaved at least about 10 times, 20 times, 30 times, 40 times, 50 times, 60 times, 70 times, 80 times, 90 times or more, or at least about 100 times faster in a target cell or under a first reference condition (which may be, e.g., selected to simulate or represent intracellular conditions) than in an individual's blood, or under a second reference condition (which may be, e.g., selected to simulate or represent conditions found in blood or serum).

[00463] Cleavable link groups are susceptible to agents of Petition 870260042207, dated 05 / 05 / 2026, page 176 / 431 160 / 386 cleavage, e.g., pH, redox potential, or the presence of degrading molecules. In general, cleavage agents are more prevalent or found at higher levels or activities than in serum or blood. Examples of such degrading agents include: redox agents that are selected for particular substrates or without substrate specificity, including, e.g., oxidizing or reducing enzymes or reducing agents such as mercaptans, present in cells, which can degrade a cleavable redox linkage group by reduction; esterases; endosomes or agents capable of creating an acidic environment, e.g., those that result in a pH equal to or less than five; enzymes capable of hydrolyzing or degrading a cleavable acid linkage group acting as an acid in general, peptidases (which may be substrate-specific), and phosphatases.

[00464] A cleavable linking group, such as a disulfide bond, may be susceptible to pH. The pH of human serum is 7.4, while the average intracellular pH is slightly lower, ranging from approximately 7.1-7.3. Endosomes have a more acidic pH, in the range of 5.5-6.0, and the pH of lysosomes is even more acidic, around 5.0. Some linkers will have a cleavable linking group that is cleaved at a preferred pH, thus releasing a cationic lipid within the cell, in the desired compartment of the cell.

[00465] A linker may include a cleavable linking group that can be cleaved by a particular enzyme. The type of cleavable linking group incorporated into the linker depends on the target cell. For example, a linker targeted to the liver may be linked to a cationic lipid via a linker that includes an ester group. Liver cells are rich in esterases, and therefore the linker will be cleaved more efficiently in liver cells than in cell types that are not rich in esterases. Other cell types rich in esterases include lung cells, renal cortex cells, and testes.

[00466] Linkers containing peptide bonds can be used Petition 870260042207, dated 05 / 05 / 2026, page 177 / 431 161 / 386 when targeting cell types rich in peptidases, such as liver cells and synoviocytes.

[00467] In general, the suitability of a candidate cleavable linking group can be assessed by testing the ability of a degrading agent (or condition) to cleave the candidate linking group. It will also be desirable to test the ability of the candidate cleavable linking group to resist cleavage in serum or when in contact with other non-target tissue. Thus, it is possible to determine the relative susceptibility to cleavage between a first and a second condition, where the first is selected to be indicative of cleavage in a target cell, and the second is selected to be indicative of cleavage in other tissues or biological fluids, e.g., blood or serum. Assessments can be performed in cell-free systems, in cells, in cell culture, in organ or tissue culture, or in whole animals. It may be useful to perform initial assessments in cell-free or culture conditions and confirm by further assessments in whole animals.In preferred embodiments, useful candidate compounds are cleaved at least approximately 2, 4, 10, 20, 30, 40, 50, 60, 70, 80, 90 or about 100 times faster in cells (or under in vitro conditions selected to simulate intracellular conditions) compared to blood or serum (or under in vitro conditions selected to simulate extracellular conditions). i. redox cleavable linking groups

[00468] In one embodiment, a cleavable linking group is a redox cleavable linking group that is cleaved upon reduction or oxidation. An example of a reduction-cleavable linking group is a disulfide linking group (-SS-). To determine whether a candidate cleavable linking group is a suitable “reduction-cleavable linking group” or, for example, whether it is suitable for use with a particular iRNA moiety and a particular targeting agent, the methods described herein can be examined. For example, a candidate can be evaluated by incubation with Petition 870260042207, dated 05 / 05 / 2026, page 178 / 431 162 / 386 dithiothreitol (DTT), or other reducing agents, using reagents known in the art, simulating the cleavage rate that would be observed in a cell, e.g., a target cell. Candidates can also be evaluated under conditions that are selected to simulate blood or serum conditions. In one embodiment, candidate compounds are cleaved by at most approximately 10% in blood. In other embodiments, useful candidate compounds are degraded at least approximately 2, 4, 10, 20, 30, 40, 50, 60, 70, 80, 90 or about 100 times faster in the cell (or under in vitro conditions selected to simulate intracellular conditions) compared to blood or serum (or under in vitro conditions selected to simulate extracellular conditions).The cleavage rate of candidate compounds can be determined by standard enzyme kinetic assays under conditions chosen to simulate the intracellular medium and compared to conditions chosen to simulate the extracellular medium. ii. Phosphate-based cleavable linking groups

[00469] In another embodiment, a cleavable linker comprises a phosphate-based cleavable linking group. A phosphate-based cleavable linking group is cleaved by agents that degrade or hydrolyze the phosphate group. An example of an agent that cleaves phosphate groups in cells are enzymes such as phosphatases in cells. Examples of phosphate-based linkage groups are -OP(O)(ORk)-O-, -OP(S)(ORk)-O-, -OP(S)(SRk)-O-, SP(O)(ORk)-O-, -OP(O)(ORk)-S-, -SP(O)(ORk)-S-, -OP(S)(ORk)-S-, -SP(S)(ORk)-O-, -OP(O)(Rk)-O-, -OP(S)(Rk)-O-, -SP(O)(Rk)-O-, -SP(S)(Rk)-O-, -SP(O)(Rk)-S-, -OP(S)(Rk)-S-. The preferred modalities are -OP(O)(OH)-O-, -OP(S)(OH)-O-, -OP(S)(SH)-O-, -SP(O)(OH)-O-, OP(O)(OH)-S-, -SP(O)(OH)-S-, -OP(S)(OH)-S-, -SP(S)(OH)-O-, -OP(O)(H)-O-, -OP(S)(H)-O-, -SP(O)(H)-O-, -SP(S)(H)-O-, -SP(O)(H)-S-, OP(S)(H)-S-. A preferred modality is -OP(O)(OH)-O-. These candidates can be evaluated using methods analogous to those described. Petition 870260042207, dated 05 / 05 / 2026, page 179 / 431 163 / 386 above. iii. Acidic cleavable linking groups

[00470] In another embodiment, a cleavable linker comprises an acidic cleavable linking group. An acidic cleavable linking group is a linking group that is cleaved under acidic conditions. In preferred embodiments, acidic cleavable linking groups are cleaved in acid with a pH near or below 6.5 (e.g., near or below 6.0; 5.75; 5.5; 5.25; 5.0), or by agents such as enzymes capable of acting as a general acid. In a cell, specific organelles with low pH, such as endosomes and lysosomes, can provide an environment for the cleavage of acidic cleavable linking groups. Examples of acidic cleavable linking groups include, but are not limited to, hydrazones, esters, and amino acid esters. Acidic cleavable groups may have the general formula -C=NN-, C(O)O, or -OC(O).A preferred embodiment is when the carbon attached to the oxygen of the ester (the alkoxy group) is an aryl group, substituted alkyl group, or tertiary alkyl group such as dimethyl, pentyl, or t-butyl. These candidates can be evaluated using methods analogous to those described above. iv. Ester-based linking groups

[00471] In another embodiment, a cleavable linker comprises an ester-based cleavable linking group. An ester-based cleavable linking group is cleaved by enzymes such as esterases and amidases in cells. Examples of ester-based cleavable linking groups include, but are not limited to, alkylene, alkenylene, and alkynylene esters. Ester-based cleavable linking groups have the general formula -C(O)O- or -OC(O)-. These candidates can be evaluated using methods analogous to those described above. v. Peptide-based cleavage groups

[00472] In yet another embodiment, a cleavable linker comprises a peptide-based cleavable linking group. A linking group Petition 870260042207, dated 05 / 05 / 2026, page 180 / 431 Peptide-based cleavable groups are cleaved by enzymes such as peptidases and proteases in cells. Peptide-based cleavable linkages are peptide bonds formed between amino acids to produce oligopeptides (e.g., dipeptides, tripeptides, etc.) and polypeptides. Peptide-based cleavable groups do not include the amide group (-C(O)NH-). The amide group can be formed between any alkylene, alkenylene, or alkynylene. A peptide bond is a special type of amide bond formed between amino acids to produce peptides and proteins. The peptide-based cleavable group is generally limited to the peptide bond (i.e., the amide bond) formed between amino acids that results in peptides and proteins and does not include the entire amide functional group. Peptide-based cleavable linking groups have the general formula -NHCHRAC(O)NHCHRBC(O)-, where RA and RB are the R groups of the two adjacent amino acids.These candidates can be evaluated using methods similar to those described above.

[00473] In one embodiment, an iRNA of the invention is conjugated to a carbohydrate via a linker. Non-limiting examples of iRNA and carbohydrate conjugated with linkers of the compositions and methods of the invention include, among others: (Formula XXXVII) THE HO / OH HoV^rAxOXz^Xz^^ AcHN OH H HO AcHN HOOH V\^O 'A.-N -OO HOOH THE HH N. / x .N (Formula XXXVIII), Petition 870260042207, dated 05 / 05 / 2026, pp. 181 / 431 165 / 386 HOOHO HO HO AcHN OH •X.QHOAcHN HOOH THE OHHOAcHN THE H NO THE NH XO OY x = 1-30 y = 1-15 (Formula XXXIX) HOOH O HO HO AcHN OH -XO HNxO OHOAcHN HOOH O OHHOAcHN HOOH O H NO O FOR H O HOAcHN HOOH O XO HN y Of xO (Formula XL) x = 1-30 y = 1-15HOAcHN HOOH OHOAcHN O O H NO S—S x x = 0-30 y = 1-15 O XO Oy (Formula XLI) HOOH OHOAcHN HOOH OHOHAcHN HOOH O OHHOAcHN O O H NO O xx = 0-30 y = 1-15 z = 1-20 XO ,,zO'Y zO Oy(Formula XLII) HOOH THE HO HO AcHN OH A-OHOAcHN HOOH THE OHHOAcHN THE THE H NO THE O' xx = 1-30 y = 1-15 z = 1-20 z O XO HN y Petition 870260042207, dated 05 / 05 / 2026, page 182 / 431 166 / 386 (Formula XLIII) and HOOH OHOAcHN HOOH THE HO AcHN HOOH OHOAcHN THE H NO THE HN THE OO' x x = 1-30 y = 1-15 z = 1-20 Oy XO (Formula XLIV), where one of X or Y is an oligonucleotide, the other is a hydrogen.

[00474] In certain embodiments of the compositions and methods of the invention, a linker is one or more GalNAc (N-acetylgalactosamine) derivatives attached via a bivalent or trivalent branched linker.

[00475] In one embodiment, a dsRNA of the invention is conjugated to a bivalent or trivalent branched linker selected from the group of structures shown in any of the formulas (XLV) - (XLVI): Formula XXXXV Formula XLVI Formula XLVII Formula XLVIII where: q2A, q2B, q3A, q3B, q4A, q4B, q5A, q5B, and q5C represent 0-20 independently for each occurrence, where the repeated unit can be the same or different; Petition 870260042207, dated 05 / 05 / 2026, p. 183 / 431 167 / 386 p2A p2B p3A p3B p4A p4B p5A p5B p5C p2A p2B p3A p3B T4A, t4b, t4a, t5b, t5cs^0, each one, independently for each occurrence, absent, CO, NH, O, S, OC(O), NHC(O), CH2, CH2NH or CH2O; Q2A, Q2B, Q3A, Q3B, Q4A, Q4B, Q5A, Q5B, Q5C are independently, for each occurrence, absent, alkylene, substituted alkylene, in which one or more methylenes may be interrupted or terminated by one or more of O, S, S(O), SO2, N(RN), C(R')=C(R''), C=C or C(O); R2A, R2B, R3A, R3B, R4A, R4B, R5A, R5B, R5C are, each independently for each occurrence absent, NH, O, S, CH2, C(O)O, O C(O)NH, NHCH(Ra)C(O), -C(O)-CH(Ra)-NH-, CO, CH=NO, heterocycline; L2A, L2B, L3A, L3B, L4A, L4B, L5A, L5B, and L5C represent the ligand; that is, each independently for each occurrence a monosaccharide (such as GalNAc), disaccharide, trisaccharide, tetrasaccharide, oligosaccharide, or polysaccharide; and RaeH or amino acid side chain. Conjugation with trivalent derivatives of GalNAc is especially use...

Claims

1 / 9 CLAIMS 1. Double-stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of 17ε-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, or a salt thereof, characterized in that the dsRNA agent, or a salt thereof, comprises a sense strand and an antisense strand forming a double-stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any of the antisense nucleotide sequences in any of Tables 2, 3, 7, 8, 10, 11 or 13.

2. A dsRNA agent, or a salt thereof, according to claim 1, characterized in that the antisense strand comprises at least 17 contiguous nucleotides of any of the antisense strand nucleotide sequences selected from the group consisting of 5'-AACAAGAUUAGUCUUGAUGUAGU-3' of SEQ ID NO:4497; 5'-AAAUUGAAUAUUCUGCAUACGAU-3' of SEQ ID NO:4501; 5'-AAAUUGAAUAUUCUGCAUACGAU-3' of SEQ ID NO:4503; 5'-ACAAGATUAGUCUUGAUGUAGUG-3' of SEQ ID NO:4505; 5'-UUAUUCCAUCUAUCAGACUUCUU-3' of SEQ ID NO:3146; 5'-AUUCCAUCUAUCAGACUUCUUAC-3' of SEQ ID NO:3145; 5'-UUCCAUCUAUCAGACUUCUUACG-3' of SEQ ID NO:3144; Petition 870260042207, dated 05 / 05 / 2026, p. 404 / 431 2 / 9 5'-UCUAUCAGACUUCUUACGACUUC-3' of SEQ ID NO:3143; 5'-UAUUCCAUCUAUCAGACUUCUUA-3' of SEQ ID NO:146; and 5'-UTCCAUCUAUCAGACUUCUUACG-3' of SEQ ID NO:1630.

3. A dsRNA agent, or a salt thereof, according to claim 1 or 2, characterized in that all nucleotides of the sense strand and all nucleotides of the antisense strand comprise a nucleotide modification.

4. A dsRNA agent, or a salt thereof, according to claim 3, characterized in that at least one of the nucleotide modifications is selected from the group consisting of a deoxynucleotide modification, a 3'-terminal deoxythymine (dT) nucleotide modification, a 2'-O-methyl nucleotide modification, a 2'-fluoro nucleotide modification, a 2'-deoxy nucleotide modification, a blocked nucleotide modification, an unblocked nucleotide modification, a conformationally restricted nucleotide modification, a constrained ethyl nucleotide modification, an abasic nucleotide modification, a 2'-amino nucleotide modification, a 2'-O-allyl nucleotide modification, a 2'-C-alkyl nucleotide modification, a 2'-hydroxyl nucleotide modification, a nucleotide modification 2'-methoxyethyl, a nucleotide modification; 2'-O-alkyl, a nucleotide modification; morpholino, a phosphoramidate modification.a nucleotide modification comprising an unnatural base, a tetrahydropyran nucleotide modification, a 1,5-anhydrohexitol nucleotide modification, a cyclohexenyl nucleotide modification, a nucleotide comprising a 5'-phosphate modification, a nucleotide comprising a 5'-phosphate mimetic modification, a glycol nucleotide modification, and a 2-O-(N-methylacetamide) nucleotide modification, and combinations thereof.

5. A dsRNA agent, or a salt thereof, according to any one of claims 1 to 4, characterized in that each strand independently has a length of 17-30 nucleotides.

6. dsRNA agent, or a salt thereof, according to claim 5, characterized in that each strand independently has a length of 19-25 nucleotides.

7. dsRNA agent, or a salt thereof, according to claim 5, characterized in that each strand independently has a length of 21-23 nucleotides.

8. A dsRNA agent, or a salt thereof, according to any one of claims 1-7, characterized in that at least one strand comprises a 3' overhang of at least 1 nucleotide.

9. A dsRNA agent, or a salt thereof, according to any one of claim 8, characterized in that at least one strand comprises a 3' overhang of at least 2 nucleotides.

10. A dsRNA agent, or a salt thereof, according to any one of claims 1 to 9, characterized in that the linker is conjugated to the 3' end of the sense strand of the dsRNA agent, or a salt thereof.

11. A dsRNA agent, or a salt thereof, according to any one of claims 1-10, characterized in that the ligand is an N-acetylgalactosamine (GalNAc) derivative.

12. dsRNA agent, or a salt thereof, according to claim 11, characterized in that the linker is linked to the dsRNA agent, or a salt thereof, by means of a monovalent linker, a bivalent linker, or a trivalent branched linker.

13. dsRNA agent, or a salt thereof, according to claim 12, characterized in that the linker is 14. dsRNA agent, or a salt thereof, according to claim 13, characterized in that the dsRNA agent, or a salt thereof, is conjugated to the ligand as shown in scheme a or S.

15. dsRNA agent, or a salt thereof, according to claim 14, characterized in that X is O.

16. dsRNA agent, or a salt thereof, according to any one of claims 1 to 15, characterized in that the dsRNA agent further comprises at least one phosphorothioate or methylphosphonate internucleotide bond. Petition 870260042207, dated 05 / 05 / 2026, p. 407 / 431 5 / 9 17. dsRNA agent, or a salt thereof, according to claim 16, characterized in that the phosphorothioate or methylphosphonate internucleotide linkage is at the 3' end of one of the strands.

18. dsRNA agent, or a salt thereof, according to claim 17, characterized in that the strand is the antisense strand.

19. dsRNA agent, or a salt thereof, according to claim 17, characterized in that the strand is the sense strand.

20. dsRNA agent, or a salt thereof, according to claim 17, characterized in that the phosphorothioate or methylphosphonate internucleotide linkage is at the 5' end of one of the strands.

21. dsRNA agent, or a salt thereof, according to claim 20, characterized in that the strand is the antisense strand.

22. dsRNA agent, or a salt thereof, according to claim 20, characterized in that the strand is the sense strand.

23. dsRNA agent, or a salt thereof, according to claim 16, characterized in that the phosphorothioate or methylphosphonate internucleotide bond is at the 5' and 3' ends of one of the strands.

24. Cell, characterized in that it contains the dsRNA agent, or a salt thereof, as defined in any one of claims 1 to 23.

25. Pharmaceutical composition for inhibiting the expression of the 17ε-hydroxysteroid dehydrogenase type 13 (HSD17B13) gene, characterized in that it comprises the dsRNA agent, or a salt thereof, as defined in any one of claims 1 to 23.

26. Pharmaceutical composition according to claim 25, characterized in that the dsRNA agent, or a salt thereof, is present in an unbuffered solution.

27. Pharmaceutical composition according to claim 26, characterized in that the unbuffered solution is saline or water. Petition 870260042207, dated 05 / 05 / 2026, pp. 408 / 431 6 / 9 28. Pharmaceutical composition according to claim 25, characterized in that the dsRNA agent, or a salt thereof, is present in a buffer solution.

29. Pharmaceutical composition according to claim 28, characterized in that said buffer solution comprises acetate, citrate, prolamine, carbonate or phosphate, or any combination thereof.

30. Pharmaceutical composition according to claim 28, characterized in that the buffer solution is phosphate-buffered saline (PBS).

31. In vitro method for inhibiting the expression of 17β-hydroxysteroid dehydrogenase type 13 (HSD17B13) in a cell, the method characterized in that it comprises contacting the cell with the agent, or a salt thereof, as defined in any one of claims 1 to 23, or with the pharmaceutical composition as defined in any one of claims 25 to 30, thereby inhibiting the expression of HSD17B13 in the cell.

32. Method according to claim 31, characterized in that the expression of HSD17B13 is inhibited by at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or below the detection level of HSD17B13 expression.

33. A dsRNA agent, or a salt thereof, according to any one of claims 1 to 23, or the pharmaceutical composition as defined in any one of claims 25 to 30, characterized in that it is for use in inhibiting the expression of HSD17B13 in an individual.

34. dsRNA agent, or a salt thereof, according to any one of claims 1 to 23, or the pharmaceutical composition as defined in any one of claims 25 to 30, characterized in that it is for use in the treatment of an individual suffering from a disease, disorder or condition associated with HSD17B13. Petition 870260042207, dated 05 / 05 / 2026, pp. 409 / 431 7 / 9 35. A dsRNA agent, or a salt thereof, according to any one of claims 1 to 23, or a pharmaceutical composition according to any one of claims 25 to 30, characterized in that it is for use in preventing at least one symptom in an individual with a disease, disorder or condition associated with HSD17B13.

36. dsRNA agent, or a salt thereof, according to any one of claims 1 to 23, or the pharmaceutical composition according to any one of claims 25 to 30, characterized in that it is for use in reducing the risk of developing chronic liver disease in an individual with steatosis.

37. dsRNA agent, or a salt thereof, according to any one of claims 1 to 23, or the pharmaceutical composition according to any one of claims 25 to 30, characterized in that it inhibits the progression of steatosis to steatohepatitis in an individual suffering from steatosis.

38. A dsRNA agent, or a salt thereof, according to any one of claims 1 to 23, or a pharmaceutical composition according to any one of claims 25 to 30 and a dsRNA agent, or a salt thereof, targeting the PNPLA3 gene, or a pharmaceutical composition comprising a dsRNA agent, or a salt thereof, targeting the PNPLA3 gene, characterized in that it inhibits the accumulation of lipid droplets in the liver of an individual suffering from a disease, disorder or condition associated with HSD17B13.

39. A dsRNA agent, or a salt thereof, according to any one of claims 1 to 23, or a pharmaceutical composition according to any one of claims 25 to 30, and a dsRNA agent, or a salt thereof, targeting the PNPLA3 gene, or a pharmaceutical composition comprising a dsRNA agent, or a salt thereof, targeting the PNPLA3 gene, characterized in that it is for use in the treatment of an individual suffering from a disease, disorder or condition associated with HSD17B13.

40. A dsRNA agent, or a salt thereof, according to any one of claims 1 to 23, or a pharmaceutical composition according to any one of claims 25 to 30, and a dsRNA agent, or a salt thereof, targeting the PNPLA3 gene, or a pharmaceutical composition comprising a dsRNA agent, or a salt thereof, targeting the PNPLA3 gene, characterized in that it is for use in preventing at least one symptom in an individual exhibiting a disease, disorder, or condition associated with HSD17B13.

41. A dsRNA agent, or a salt thereof, according to any one of claims 1 to 23, or a pharmaceutical composition according to any one of claims 25 to 30, and a dsRNA agent, or a salt thereof, targeting the PNPLA3 gene, or a pharmaceutical composition comprising a dsRNA agent, or a salt thereof, targeting the PNPLA3 gene, characterized in that it is for use in reducing the risk of developing chronic liver disease in an individual with steatosis.

42. A dsRNA agent, or a salt thereof, according to any one of claims 1 to 23, or a pharmaceutical composition according to any one of claims 25 to 30, and a dsRNA agent, or a salt thereof, targeting the PNPLA3 gene, or a pharmaceutical composition comprising a dsRNA agent, or a salt thereof, targeting the PNPLA3 gene, characterized in that it is for use in inhibiting the progression of steatosis to steatohepatitis in an individual suffering from steatosis.

43. dsRNA agent, or a salt thereof, or the pharmaceutical composition according to any of claims 34, 35 and 39 to 41, characterized in that the disease, disorder or condition associated with Petition 870260042207, dated 05 / 05 / 2026, page 411 / 431 9 / 9 HSD17B13 is a chronic fibro-inflammatory liver disease.

44. dsRNA agent, or a salt thereof, or the pharmaceutical composition according to claim 43, characterized in that chronic fibro-inflammatory liver disease is associated with the accumulation and / or expansion of lipid droplets in the liver.

45. dsRNA agent, or a salt thereof, or the pharmaceutical composition according to claim 44, characterized in that it is selected from the group consisting of liver fat accumulation, liver inflammation, hepatic fibrosis, fatty liver disease (steatosis), non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), liver cirrhosis, alcoholic steatohepatitis (ASH), alcoholic liver disease (ALD), HCV-associated cirrhosis, drug-induced liver injury, and hepatocellular necrosis.

46. ​​dsRNA agent, or a salt thereof, or the pharmaceutical composition according to claim 44, characterized in that the chronic fibro-inflammatory liver disease is non-alcoholic steatohepatitis (NASH).

47. dsRNA agent, or a salt thereof, or the pharmaceutical composition according to any one of claims 33 to 46, characterized in that the individual is obese.

48. dsRNA agent, or a salt thereof, or the method of pharmaceutical composition according to any of claims 33 to 47, characterized in that the use further comprises the use of an additional therapeutic agent.

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