Double stranded rnai agents, compositions and methods of use

Abstract

Disclosed are, inter alia, double stranded RNAi (dsRNAi) agents inhibiting expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), for example, human HMGCR, compositions including the same, and methods of treatment using the same.

Description

PAT059649-WO-PCT DOUBLE STRANDED RNAi AGENTS, COMPOSITIONS AND METHODS OF USE CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims priority and benefit to the U.S. Patent Application No. 63 / 670,342 filed July 12, 2024 and U.S. Patent Application No.63 / 786,760 filed April 10, 2025, the disclosure of each of which is incorporated herein by reference in its entirety. TECHNICAL FIELD

[0002] The present disclosure provides, inter alia, double stranded RNAi (dsRNAi) agents inhibiting expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), for example, human HMGCR, compositions including the same, and methods of treatment using the same. BACKGROUND

[0003] Cumulative low-density lipoprotein cholesterol (LDL-C) exposure in the arterial wall is a major cause of atherosclerotic cardiovascular disease (ASCVD). The level of LDL-C in the arterial wall can be controlled (e.g., lowered) by modulating cholesterol homeostasis (e.g., cholesterol biosynthesis in liver cells) and upregulating LDL-C uptake from the blood.

[0004] 3-Hydroxy-3-methylglutaryl-CoA reductase (HMGCR) is a key enzyme that converts 3- hydroxy-3-methyl-glutaryl-CoA (HMGCoA) into mevalonate in the rate-limiting step in the cholesterol biosynthesis pathway. Due to its critical role in cholesterol biosynthesis, HMGCR has been a target for drug development for the treatment of high cholesterol and cardiovascular disease risk reduction (CVRR). For example, statins, small molecule inhibitors of HMGCR, are the current standard of care for lowering cholesterol. However, statins also have adverse side effects such as myalgia and rhabdomyolysis, a rare, but potentially fatal, breakdown of skeletal muscle. Real-world studies demonstrate that a staggering 80% of US patients on standard of care do not achieve their LDL-C goal. This is due, in large part, to poor adherence.

[0005] Therefore, there is an unmet need in the art for alternative treatments (e.g., lowering cholesterol or LDL-C) for subjects having lipid metabolism disorders. SUMMARY OF THE INVENTION

[0006] Provided herein are, inter alia, compounds that can inhibit expression of HMGCR in a subject, for example, e.g., in liver cells of a subject.

[0007] In an aspect, provided is a double stranded RNAi (dsRNAi) agent comprising: 1PAT059649-WO-PCT a sense strand comprising a nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440 in Table 1; and an antisense strand forming a duplex with the sense strand and comprising a nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447 in Table 1.

[0008] In some embodiments, the sense strand is 21 to 23 nucleotides in length and the antisense strand is 23 to 25 nucleotides in length.

[0009] In some embodiments, all the nucleotides in the sense strand and the antisense strand are modified nucleotides.

[0010] In some embodiments, each of the modified nucleotides independently comprises one or more modifications selected from a 2′-deoxy modification, a 2′-O-alkyl modification, a 2′-halo modification, a threofuranosyl nucleotide (TNA) modification, a 2′-5′-linkage modification, a conformationally restricting modification, an abasic modification, a 2′- amino-modification, a 2′-O-allyl modification, 2′-C-alkyl modification, a 2′-O-alkoxyalkyl modification, a morpholino modification, a phosphoramidate modification, a non-natural nucleobase modification, a modification in a tetrahydropyran, a modification containing a 1,5-anhydrohexitol, a modification containing a cyclohexenyl, a modification containing a phosphorothioate group, a modification containing a 5′-vinyl-phosphonate, a modification containing a 5′-phosphate, a modification to form a thermally destabilizing nucleotide, a glycol nucleic acid (GNA) modification, and a 2-O-(N-methylacetamide) modification.

[0011] In some embodiments, each of the modified nucleotides independently comprises one or more modifications selected from 2′-deoxy modification, 2′-O-alkoxyalkyl modification, 2′-O-alkyl modification, 2′-O-allyl modification, 2′-C-allyl modification, 2′- halo modification, modification containing a non-natural nucleobase, GNA modification, and TNA modification.

[0012] In some embodiments, all the modified nucleotides comprise a modification on a 2′ sugar ring.

[0013] In some embodiments, the modified nucleotides are selected from a 2′-O-alkyl modified nucleotide, a 2′-halo modified nucleotide, a 2′-deoxy modified nucleotide, a 2′-O- alkoxyalkyl modified nucleotide and TNA modification.

[0014] In some embodiments, one or more of the modified nucleotides further comprises a 3′-phosphorothioate (PS) modification.

[0015] In some embodiments, each of the modified nucleotides independently comprises one or more modifications selected from 2′-deoxy modification, 2′-O-methyl (2′-OMe) 2PAT059649-WO-PCT modification, 2′-fluoro (2′-F) modification, 2′-O-methoxyethyl (2′-MOE) modification, the modification containing a non-natural nucleobase, TNA, GNA, 3′-phosphorothioate (PS) modification, and 5′-vinyl-phosphonate (5′-VP) modification.

[0016] In some embodiments, the sense strand comprises one or two 2′-MOE modified nucleotides positioned at the 1stand / or 2ndnucleotides from the 5′-end of the sense strand.

[0017] In some embodiments, the sense strand comprises one or two 2′-MOE modified nucleotides positioned at the 1stand / or 2ndnucleotides from the 3′-end of the sense strand.

[0018] In some embodiments, the sense strand comprises one or two TNAs positioned at the 1stand / or 2ndnucleotides from the 5′-end of the sense strand.

[0019] In some embodiments, the sense strand comprises one or two TNAs positioned at the 1stand / or 2ndnucleotides from the 3′-end of the sense strand.

[0020] In some embodiments, the antisense strand comprises a 5′-VP group at the 1stnucleotide from 5′ end of the antisense strand.

[0021] In some embodiments, the antisense strand comprises a 5′-(E)-VP group at the 1stnucleotide from 5′ end of the antisense strand.

[0022] In some embodiments, the antisense strand comprises a 5′-(E)-VP-2′-OMe nucleotide at the 1stposition from 5′ end of the antisense strand.

[0023] In some embodiments, each of the sense strand and the antisense strand independently comprises two, three, four, five or six 2′-F modified nucleotides.

[0024] In some embodiments, the sense strand comprises one or two 3′-PS group at the 1stand / or 2ndnucleotides from 5′-end of the sense strand.

[0025] In some embodiments, the antisense strand comprises one or two 3′-PS group at the 1stand / or 2ndnucleotides from 5′-end of the antisense strand, and / or one or two 3′-PS group at the 1stand / or 2ndnucleotides from 3′-end of the antisense strand.

[0026] In some embodiments, the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.

[0027] In some embodiments, the sense strand comprises one to four 2′-MOE modified nucleotides positioned at the 1st, 2nd, 20th, and / or 21stnucleotides from the 5′-end of the sense strand.

[0028] In some embodiments, the sense strand comprises only four 2′-MOE modified nucleotides.

[0029] In some embodiments, the sense strand does not comprise a 2′-MOE modified nucleotide at the 3rdto 19thpositions from 5′-end of the sense strand. 3PAT059649-WO-PCT

[0030] In some embodiments, the sense strand comprises one to four TNAs positioned at the 1st, 2nd, 20th, and / or 21stnucleotides from the 5′-end of the sense strand.

[0031] In some embodiments, the sense strand comprises two, three, or four 2′-F modified nucleotides positioned at the 7th, 9th, 10th, and / or 11thnucleotide from 5′-end of the sense strand.

[0032] In some embodiments, the sense strand comprises 2′-F modified nucleotides positioned at the 7th, 9th, 10th, and 11thnucleotides from 5′-end of the sense strand.

[0033] In some embodiments, the remaining nucleotides in the sense strand comprise 2′- OMe modified modification.

[0034] In some embodiments, the antisense strand comprises a 5′-(E)-VP group at the 1stnucleotide from 5′ end of the antisense strand.

[0035] In some embodiments, the antisense strand comprises two, three, or four 2′-F modified nucleotides positioned at the 2nd, 6th, 14th, and / or 16thnucleotides from 5′-end of the antisense strand.

[0036] In some embodiments, the antisense strand comprises 2′-F modified nucleotides positioned at the 2nd, 6th, 14th, and 16thnucleotides from 5′-end of the antisense strand.

[0037] In some embodiments, the antisense strand comprises 2′-F modifications positioned at the 2nd, 6th, 14th, and 16th nucleotides from the 5′ end; and (i) a GNA positioned at the 5thnucleotide from 5′ end, or (ii) a TNA positioned at the 3rd nucleotide from the 5′ end.

[0038] In some embodiments, the remaining nucleotides in antisense strand comprise 2′- OMe modified modifications.

[0039] In some embodiments, the sense strand comprises one to eight 3′-PS group at the 1st, 2nd, 3rd, 4th, 17th, 18th, 19thand / or 20thnucleotides from 5′-end of the sense strand.

[0040] In some embodiments, the antisense strand comprises one to eight 3′-PS group at the 1st, 2nd, 3rd, 4th, 19th, 20th, 21stand / or 22ndnucleotides from 5′-end of the antisense strand.

[0041] In some embodiments, at least one of the 3′-PS groups in each sense strand and antisense strand has a stereopure Rp configuration.

[0042] In some embodiments, at least one of the 3′-PS groups in each sense strand and antisense strand has a stereopure Sp configuration.

[0043] In certain aspects, provided is a double stranded RNAi (dsRNAi) agent comprising: 4PAT059649-WO-PCT a sense strand having a nucleotide sequence selected from SEQ ID NOs: 812 to 1052 in Table 2 and SEQ ID NOs: 1294 to 1297, 1448 to 1462, and 1481 to 1482 in Table 3; and an antisense strand forming a duplex with the sense strand and having a nucleotide sequence selected from SEQ ID NOs: 1053 to 1293 in Table 2 and 1298 to 1301, 1463 to 1477, and 2600 to 2605 in Table 3.

[0044] In some embodiments, the dsRNAi comprises a ligand.

[0045] In some embodiments, the ligand comprises a N-acetylgalactosamine (GalNAc) moiety.

[0046] In some embodiments, the ligand has a structure of: ,each L1is independently a linker which may be same or different in each occurrence; L2is a linker; n is an integer from 1 to 3; and is an attachment point to the sense strand or an antisense strand.

[0047] In some embodiments, the ligand comprises the following structure of , or5PAT059649-WO-PCT ,each p1, p2, p3, q1, q2, r1, r2 and r3 is independently an integer from 0 to 12; each n1, n2, and n3 is independently an integer from 1 to 3; and “*” is an attachment point to L2.

[0048] In some embodiments, the ligand has a structure of: F),each L11, L12, L13, L14, and L15is an independently a linker; L2is a linker; is an attachment point to the sense strand or the antisense strand.

[0049] In some embodiments, the ligand has a structure of: 6PAT059649-WO-PCT 1), 2),wherein: each p11 and q11 is independently an integer from 0 to 12; each z1, z2, and z3 is independently an integer of 0 to 12; and is an attachment point to the sense strand or the antisense strand.

[0050] In some embodiments, the ligand comprises the following structure: ,7PAT059649-WO-PCT ,is an attachment point to the sense strand or the antisense strand.

[0051] In some embodiments, the ligand is conjugated to 3′ end of the sense strand to form the following structure: 8PAT059649-WO-PCTwherein W is -OH or -SH.

[0052] In some embodiments, the ligand is conjugated to 5′ end of the sense strand to form the following structure: ,p y p , wherein W is -OH or -SH.

[0053] In some embodiments, W is -OH. 9PAT059649-WO-PCT

[0054] In some embodiments, the dsRNAi agent is in a pharmaceutically acceptable salt form.

[0055] In some embodiments, the pharmaceutically acceptable salt is a sodium salt.

[0056] In certain aspects, provided is a pharmaceutical composition comprising the dsRNAi agent as described herein, and a pharmaceutically acceptable carrier.

[0057] In some embodiments, the composition is in an aqueous solution form.

[0058] In some embodiments, the pharmaceutical composition further comprises an additional therapeutic agent selected from a proprotein convertase subtilisin kexin 9 (PCSK9) inhibitor, a lysophosphatidic acid (LPA) receptor inhibitor, an angiotensinogen (AGT) inhibitor, a fibrate, a bile acid sequestrant, niacin, an antiplatelet agent, an angiotensin converting enzyme inhibitor, an angiotensin II receptor antagonist, an acylCoA cholesterol acetyltransferase (ACAT) inhibitor, a cholesterol absorption inhibitor, a cholesterol ester transfer protein (CETP) inhibitor, a microsomal triglyceride transfer protein (MTTP) inhibitor, a cholesterol modulator, a bile acid modulator, a peroxisome proliferation activated receptor (PPAR) agonist, a gene-based therapy, a composite vascular protectant, a glycoprotein IIb / IIIa inhibitor, aspirin or an aspirin-like compound, an IBAT inhibitor, a squalene synthase inhibitor, a monocyte chemoattractant protein (MCP)-I inhibitor, and a combination thereof.

[0059] In some embodiments, the additional therapeutic agent comprises the PCSK9 inhibitor.

[0060] In some embodiments, the PCSK9 inhibitor is a second dsRNAi agent.

[0061] In some embodiments, the second dsRNAi agent comprises inclisiran.

[0062] In certain aspects, provided is a combination of the dsRNAi agent as described herein and a second agent selected from a proprotein convertase subtilisin kexin 9 (PCSK9) inhibitor, a lysophosphatidic acid (LPA) receptor inhibitor, an angiotensinogen (AGT) inhibitor, a fibrate, a bile acid sequestrant, niacin, an antiplatelet agent, an angiotensin converting enzyme inhibitor, an angiotensin II receptor antagonist, an acylCoA cholesterol acetyltransferase (ACAT) inhibitor, a cholesterol absorption inhibitor, a cholesterol ester transfer protein (CETP) inhibitor, a microsomal triglyceride transfer protein (MTTP) inhibitor, a cholesterol modulator, a bile acid modulator, a peroxisome proliferation activated receptor (PPAR) agonist, a gene-based therapy, a composite vascular protectant, a glycoprotein IIb / IIIa inhibitor, aspirin or an aspirin-like compound, an IBAT inhibitor, a 10PAT059649-WO-PCT squalene synthase inhibitor, a monocyte chemoattractant protein (MCP)-I inhibitor, and a combination thereof.

[0063] In some embodiments, the second agent is a second dsRNAi agent.

[0064] In some embodiments, the second dsRNAi agent is a dsRNA agent that targets one or more of the genes selected from the group consisting of PCSK9, LPA, AGT, ACE, ACE2, AGTR1, AGTR2, ACAT, CETP, MTTP, PPAR, IBAT, FDFT1, ERG9, SQS1, Ccl2, CCR2, CCL7, CCL8. CCL13, and CCL16.

[0065] In some embodiments, the second dsRNAi agent comprises inclisiran.

[0066] In certain aspects, provided is a pharmaceutical composition comprising the combination as described herein.

[0067] In some embodiments, the second dsRNAi agent is in a pharmaceutically acceptable salt form.

[0068] In some embodiments, the pharmaceutically acceptable salt of the second dsRNAi agent is a sodium salt.

[0069] In some embodiments, the dsRNAi agent and the second agent are formulated in the same composition.

[0070] In some embodiments, the dsRNAi agent and the second agent are formulated in the separate compositions.

[0071] In certain aspects, provided is a method of inhibiting expression of 3-hydroxy-3- methylglutaryl-CoA reductase (HMGCR) in a subject comprising: administering to the subject the dsRNAi agent as described herein or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described herein.

[0072] In certain aspects, provided is method of lowering a level of low-density lipoprotein cholesterol (LDL-C) in a subject, comprising: administering to the subject the dsRNAi agent as described herein or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described herein.

[0073] In certain aspects, provided is a method of treating or preventing an HMGCR- associated disorder or disease in a subject, comprising: administering to the subject the dsRNAi agent as described herein or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described herein. 11PAT059649-WO-PCT

[0074] In some embodiments, the HMGCR-associated disorder or disease is hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, primary hyperlipidemia, heterozygous familiar hypercholesterolemia (HeFH), homozygous familiar hypercholesterolemia (HoFH), congestive heart disease (CHD) or atherosclerosis.

[0075] In certain aspects, provided is a method of treating or preventing hyperlipidemia in a subject, comprising: administering to the subject the dsRNAi agent as described herein or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described herein.

[0076] In some embodiments, the hyperlipidemia is hypercholesterolemia, or hypertriglyceridemia.

[0077] In certain aspects, provided is a method of treating or preventing atherosclerotic cardiovascular disease (ASCVD) in a subject, comprising: administering to the subject the dsRNAi agent as described herein or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described herein.

[0078] In some embodiments, the dsRNAi agent or the pharmaceutical composition is administered subcutaneously or intravenously.

[0079] In some embodiments, the methods further comprises administering to the subject an additional therapeutic agent selected from a proprotein convertase subtilisin kexin 9 (PCSK9) inhibitor, a lysophosphatidic acid (LPA) receptor inhibitor, an angiotensinogen (AGT) inhibitor, a fibrate, a bile acid sequestrant, niacin, an antiplatelet agent, an angiotensin converting enzyme inhibitor, an angiotensin II receptor antagonist, an acylCoA cholesterol acetyltransferase (ACAT) inhibitor, a cholesterol absorption inhibitor, a cholesterol ester transfer protein (CETP) inhibitor, a microsomal triglyceride transfer protein (MTTP) inhibitor, a cholesterol modulator, a bile acid modulator, a peroxisome proliferation activated receptor (PPAR) agonist, a gene-based therapy, a composite vascular protectant, a glycoprotein IIb / IIIa inhibitor, aspirin or an aspirin-like compound, an IBAT inhibitor, a squalene synthase inhibitor, a monocyte chemoattractant protein (MCP)-I inhibitor, and a combination thereof.

[0080] In some embodiments, the additional therapeutic agent is a second dsRNAi agent.

[0081] In some embodiments, the second dsRNAi agent comprises the PCSK9 inhibitor.

[0082] In some embodiments, the second dsRNAi agent comprises inclisiran. 12PAT059649-WO-PCT

[0083] In some embodiments, the dsRNAi agent or the pharmaceutical composition and the additional therapeutic agent are administered simultaneously.

[0084] In some embodiments, the dsRNAi agent or the pharmaceutical composition and the additional therapeutic agent are administered subsequently.

[0085] In some embodiments, the dsRNAi agent is administered before administering the additional therapeutic agent.

[0086] In some embodiments, the additional therapeutic agent is administered before administering the dsRNAi agent.

[0087] In some embodiments, the additional therapeutic agent is administered subcutaneously or intravenously.

[0088] In certain aspects, provided is a method of lowering a level of low-density lipoprotein cholesterol (LDL-C) in a subject, comprising: administering to the subject the pharmaceutical composition as described herein.

[0089] In certain aspects, provided is a method of treating or preventing an HMGCR- associated disorder or disease in a subject, comprising: administering to the subject the pharmaceutical composition as described herein.

[0090] In some embodiments, the HMGCR-associated disorder or disease is hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, primary hyperlipidemia, heterozygous familiar hypercholesterolemia (HeFH), homozygous familiar hypercholesterolemia (HoFH), congestive heart disease (CHD) or atherosclerosis.

[0091] In certain aspects, provides is a method of treating or preventing hyperlipidemia in a subject, comprising: administering to the subject the pharmaceutical composition as described herein.

[0092] In certain aspects, provided is a method of treating or preventing atherosclerotic cardiovascular disease (ASCVD) in a subject, comprising: administering to the subject the pharmaceutical composition as described herein.

[0093] In some embodiments, the dsRNAi agent and the second agent is administered subcutaneously or intravenously.

[0094] In some embodiments, the dsRNAi agent and the second agent are administered simultaneously.

[0095] In some embodiments, the dsRNAi agent and the second agent are administered subsequently. 13PAT059649-WO-PCT

[0096] In some embodiments, the dsRNAi agent is administered before administering the second agent.

[0097] In some embodiments, the second agent is administered before administering the dsRNAi agent.

[0098] In some embodiments, in any of the methods described herein, the subject is a human.

[0099] In some embodiments, in any of the methods described herein, the subject has or is diagnosed with hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, primary hyperlipidemia, heterozygous familiar hypercholesterolemia (HeFH), homozygous familiar hypercholesterolemia (HoFH), congestive heart disease (CHD) or atherosclerosis.

[0100] In some embodiments, in any of the methods described herein, the subject does not have a muscle side effect after the administrating the pharmaceutical composition of as described herein.

[0101] In certain aspects, provided is a method of reducing the risk of a major adverse cardiovascular event in a subject, comprising administering to the subject the dsRNAi agent as described herein or a pharmaceutically acceptable salt thereof, the pharmaceutical composition as described herein, the combination as described herein, or the pharmaceutical composition comprising the combination as described herein.

[0102] In some embodiments, the major adverse cardiovascular event is cardiovascular death, non-fatal myocardial infarction, non-fatal ischemic stroke, or urgent coronary revascularization.

[0103] In some embodiments, the subject has an established cardiovascular disease.

[0104] In some embodiments, the subject has not experienced a major atherosclerotic cardiovascular disease (ASCVD) event.

[0105] In certain aspects, provided is a kit comprising the dsRNAi agent as described herein or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described herein.

[0106] In some embodiments, the kit further comprises an additional therapeutic agent selected from a proprotein convertase subtilisin kexin 9 (PCSK9) inhibitor, a fibrate, a bile acid sequestrant, niacin, an antiplatelet agent, an angiotensin converting enzyme inhibitor, an angiotensin II receptor antagonist, an acylCoA cholesterol acetyltransferase (ACAT) inhibitor, a cholesterol absorption inhibitor, a cholesterol ester transfer protein (CETP) 14PAT059649-WO-PCT inhibitor, a microsomal triglyceride transfer protein (MTTP) inhibitor, a cholesterol modulator, a bile acid modulator, a peroxisome proliferation activated receptor (PPAR) agonist, a gene-based therapy, a composite vascular protectant, a glycoprotein IIb / IIIa inhibitor, aspirin or an aspirin-like compound, an IBAT inhibitor, a squalene synthase inhibitor, a monocyte chemoattractant protein (MCP)-I inhibitor, and a combination thereof.

[0107] In some embodiments, the additional therapeutic agent is a second dsRNAi agent.

[0108] In some embodiments, the second dsRNAi agent is a dsRNA agent that targets one or more of the genes selected from the group consisting of PCSK9, LPA, AGT, ACE, ACE2, AGTR1, AGTR2, ACAT, CETP, MTTP, PPAR, IBAT, FDFT1, ERG9, SQS1, Ccl2, CCR2, CCL7, CCL8. CCL13, and CCL16.

[0109] In some embodiments, the second dsRNAi agent comprises the PCSK9 inhibitor.

[0110] In some embodiments, the second dsRNAi agent comprises inclisiran.

[0111] In some embodiments, the dsRNAi agent and the additional therapeutic agent are contained in a single vial.

[0112] In some embodiments, the dsRNAi agent and the additional therapeutic agent are contained in separate vials.

[0113] In certain aspects, provided is a kit comprising the pharmaceutical composition including the combination as described herein.

[0114] In some embodiments, the dsRNAi agent and the second agent are contained in a single vial.

[0115] In some embodiments, the dsRNAi agent and the second agent are contained in separate vials.

[0116] In some embodiments, the kir further comprises one or more applicators.

[0117] In some embodiments, the one or more applicators comprises a syringe.

[0118] Other aspects of the invention are disclosed infra. BRIEF DESCRIPTION OF DRAWINGS

[0119] Figures 1A to 1C: Transcriptomic profiling with different lead sequences. The siRNA284, for example, which can be formed by SEQ ID NO: 284 and SEQ ID NO: 689, or modified variants thereof had HMGCR significant down-regulation as shown in Figure 1A. The siRNA3, for example, which can be formed which can be formed by SEQ ID NO: 3 and SEQ ID NO: 408, or modified variants thereof had an exquisite off-target profile as shown in Figure 1B. The siRNA6, for example, which can be formed by SEQ ID NO: 6 and SEQ ID 15PAT059649-WO-PCT NO: 411, or modified variants thereof (e.g., with or without GNA) had still some seed- mediated off-targeting profiling as shown in Figure 1C.

[0120] Figure 2: Compounds 1-4 as outlined in Table 5 of the disclosure are depicted and each modification (e.g., 2′ modified nucleosides and linkages) are depicted. “PO” means a chemical group to form a phosphate (phosphodiester) linkage and “PS” means linking group to form a phosphorothioate linkage, and “L96” refers to a ligand that is connected to the 3′-end of the sense strand via phosphate (phosphodiester) linkage. Figure discloses SEQ ID NOS 2606-2613, respectively, in order of appearance.

[0121] Figures 3A to 3B: Liver HMGCR mRNA concentrations for male C57BL / 6 mice (n=4 per timepoint for each group) administered a single subcutaneous dose of either Compound 7 (gray) or Compound 6 (black) at 6 mg / kg are shown in Figure 3A. Statistical significance was determined by ordinary one-way ANOVA and Sidak′s multiple comparisons test. For Compound 7, statistically significant reductions in HMGCR mRNA abundance versus vehicle were observed at all post-dose timepoints, with the exception of day 28. A maximum mean reduction of 77±3% (P<0.001) occurred on day 21 and was, in large part, sustained through day 35 (-68±12%, P<0.01). Compound 6 also reduced liver HMGCR mRNA abundance in this model, with a maximum decrease of 79±12% at day 7. Incorporation of guide strand into liver RISC over time is illustrated in Figure 3B. Statistical significance was determined by ordinary one-way ANOVA and Sidak′s multiple comparisons test. In male C57BL / 6 mice (n=4 per timepoint for each group) dosed with 6 mg / kg Compound 6 (black circles), a steep decline in RISC-loading was observed between days 7 and 21 post-dose. In contrast, RISC-loading was sustained through day 21 in mice administered Compound 7 at 6 mg / kg (gray squares). Moreover, at day 42 post-dose, RISC- loading was 100-fold greater in the Compound 7 versus Compound 6 group.

[0122] Figures 4A to 4B: Liver HMGCR mRNA concentrations for male C57BL / 6 mice (n=4 or 5 per group) administered a single subcutaneous dose of PBS (white), Compound 5 (white / black), Compound 7 (black) or Compound 8 (gray) at 3 mg / kg are shown in Figure 4A. The siRNAs share a same nucleotide sequence but have different chemical modifications. Statistical significance was determined by ordinary one-way ANOVA and Sidak′s multiple comparisons test. At day 35 post-dose, statistically significant reductions in hepatic HMGCR mRNA abundance versus vehicle (PBS) were observed in mice dosed with Compound 7 (- 55%, P<0.05 vs. PBS) and Compound 8 (-70%, P<0.01 vs. PBS). In mice dosed with Compound 8, liver HMGCR mRNA levels were significantly reduced vs. vehicle through day 16PAT059649-WO-PCT 56 post-dose (-51%, P<0.05). These results show durable HMGCR knockdown in mice, with Compound 8 performing better than Compound 7 in this experiment. Incorporation of guide strand into liver RISC over time is illustrated in Figure 4B. Male C57BL / 6 mice (n= 4 or 5 per group) were administered a single subcutaneous dose of PBS (white) or Compound 5 (white / black), Compound 7 (black) or Compound 8 (gray) at 3 mg / kg. Statistical significance was determined by ordinary one-way ANOVA and Sidak′s multiple comparisons test. Consistent with the liver HMGCR mRNA results, Compound 8 showed the highest RISC- loading at both timepoints evaluated. The difference observed between Compound 5 and Compound 8 at day 35 was robust and statistically significant (P<0.0001). These results show durable RISC-incorporation (through day 56 post-dose) for Compound 8 in mice.

[0123] Figures 5A to 5B: Nine- to ten-week old male Wistar Han rats received a single subcutaneous dose of vehicle (0.9% sodium chloride for injection, USP) or HMGCR GalNAc-conjugated siRNA (Compound 9) on study day 1. Livers for the measurement of HMGCR mRNA abundance were collected at necropsy (30 days post-dose). HMGCR mRNA levels are normalized to TATA-box binding protein (TBP) and represent mean ± SD (n=5 per group) in Figure 5A. Statistical significance was determined by ordinary one-way ANOVA and Dunnett′s multiple comparisons test. Statistically significant reductions in hepatic HMGCR mRNA abundance versus vehicle were observed for all siRNA groups. Maximum target knockdown was achieved with doses of ≥30 mg / kg. Nine- to ten-week old male Wistar Han rats received a single subcutaneous dose of vehicle (0.9% sodium chloride for injection, USP) or HMGCR GalNAc-conjugated siRNA (Compound 9) on study day 1. Right bicep femoris skeletal muscle samples for the measurement of HMGCR mRNA abundance were collected at necropsy (30 days post-dose). Mean HMGCR mRNA abundance for vehicle and GalNAc-conjugated HMGCR siRNA (Compound 9) groups are shown in Figure 5B and represent mean ± SD (n=5 per group). Relative to vehicle, skeletal muscle HMGCR mRNA levels were minimally increased in all dose groups, with no dose-relatedness evident. None of these differences achieved statistical significance, when compared to the vehicle group. These findings demonstrate that, at doses up to 300 mg / kg, target knockdown is not observed in skeletal muscle 30 days post-dose.

[0124] Figures 6A to 6B: Low density lipoprotein receptor (LDLR) protein levels in a human liver cell line (Huh7) treated with HMGCR siRNA (Compound 1) or atorvastatin at a concentration of 25 nM are provided in Figure 6A. Briefly, Huh7 cells were transfected with test or control siRNA using lipofectamine and incubated at 37°C for 6 hours. After changing 17PAT059649-WO-PCT the culture media, cells were incubated for another 24 hours prior to processing for the measurement of mRNA abundance or LDLR protein levels. The same incubation times were used for cells treated with atorvastatin. Atorvastatin (25 µM in DMSO) was diluted in culture medium to achieve a concentration of 25 nM. HMGCR mRNA was reduced by 61% versus control in Huh7 cells treated with HMGCR siRNA (Compound 1). Total LDLR protein levels, as determined by Western blotting, increased by 57% or 20% in cells treated with HMGCR siRNA (Compound 1) or atorvastatin, respectively. These results demonstrate that, like a statin, an HMGCR siRNA (Compound 1) increases LDLR protein in a human liver cell line. LDLR protein levels in primary human hepatocytes (PHH) treated with HMGCR siRNA (Compound 1) or atorvastatin at a concentration of 10 nM are provided in Figure 6B. Briefly, PHH were transfected with test or control siRNA using lipofectamine and incubated at 37°C for 24 hours prior to processing for the measurement of mRNA abundance or LDLR protein levels. The same incubation times were used for cells treated with atorvastatin. Atorvastatin (25 µM in DMSO) was diluted in culture medium to achieve a concentration of 10 nM. HMGCR mRNA was reduced by 65% versus control in PHH cells treated with HMGCR siRNA (Compound 1). Total LDLR protein levels, as determined by Western blotting, increased by 48% or 80% in cells treated with HMGCR siRNA (Compound 1) or atorvastatin, respectively. These results demonstrate that, like a statin, an HMGCR siRNA (Compound 1) increases LDLR protein in primary human hepatocytes.

[0125] Figures 7A-7B: HMGCR protein levels in a human liver cell line (Huh7) treated with Compound 1 at a concentration of 12.5 or 25 nM are provided in Figure 7A. Briefly, Huh7 cells were transfected with test or control siRNA using lipofectamine and incubated at 37°C for 6 hours. After changing the culture media, cells were incubated for another 24 hours prior to processing for the measurement of HMGCR protein levels by Western blotting. Relative to the control siRNA, Compound 1 reduced HMGCR protein content by 51% and 73% at concentrations of 12.5 and 25 nM, respectively. These results demonstrate that Compound 1 significantly, and dose-dependently, reduces HMGCR protein content in a human liver cell line. HMGCR protein levels in a human liver cell line (Huh7) treated with atorvastatin at a concentration of 12.5 or 25 nM are provided in Figure 7B. Atorvastatin (25 µM in DMSO) was diluted in culture medium to achieve the desired concentrations. Huh7 cells were incubated with atorvastatin or DMSO at 37°C for 6 hours. After changing the culture media, cells were incubated for another 24 hours prior to processing for the measurement of HMGCR protein levels by Western blotting. Relative to DMSO, atorvastatin 18PAT059649-WO-PCT markedly augmented HMGCR protein levels, with 96% and 123% increases observed relative to control at concentrations of 12.5 and 25 nM, respectively. These results demonstrate that, in contrast to Compound 1, atorvastatin robustly increases HMGCR protein expression in a human liver cell line.

[0126] Figure 8A-8B: Effects of Compound 1 on plasma total cholesterol concentrations in mice with humanized livers. Mean percent change ± SD in total cholesterol level versus baseline for the PBS and Compound 1 groups are shown in Figure 8A, while results for individual animals in the Compound 1 group are presented in panel Figure 8B. TC, total cholesterol.

[0127] Figures 9A-9B: Hepatic HMGCR mRNA abundance in male cynomolgus monkeys at pre-dose and days 28 and 85 after subcutaneous injection of two doses of Compound 1 given 34 days apart. A total of 3 liver biopsy samples were obtained from each animal during the study: one pre-dose (day 12) and two post-dose (days 28 and 85 after the first dose) to enable the measurement of hepatic HMGCR mRNA expression by RT-qPCR. HMGCR mRNA levels are normalized to TATA-box binding protein (TBP) and represent mean ± SD (n=4 per group). Shapes indicate values for individual animals in each group. Statistical significance was determined by paired t-test. Mean hepatic HMGCR mRNA abundance was reduced versus pre-dose in all groups at both day 28 and 85; however, only the Compound 1, 5 mg / kg group, showed statistically significant reductions in target knockdown at both timepoints. *P<0.0003, **P<0.05 for the comparison with pre-dose values.

[0128] Figure 10: Serum HDL cholesterol concentrations in male cynomolgus monkeys at post-dose timepoints through day 85 after subcutaneous injection of two doses of Compound 1 given 34 days apart. On days 1 and 35, male cynomolgus monkeys (n=4 per group) received Compound 1 at 5 or 10 mg / kg via subcutaneous injection. Blood samples were collected at 3 pre-dose timepoints and at multiple post- dose timepoints for the determination of serum HDL cholesterol levels, using a Cobas 8000 chemistry analyzer. Data represent mean ± SD percent change versus pre-dose (average of measurements conducted on samples from 3 pre-dose blood collections) through day 85 (day of final liver biopsy) post initial dose.

[0129] Figure 11: Dose response curve in Hep3B cells transfected with nine siRNAs (S1-S9) of Example 10 in an 8-point dilution series starting at 40 nM with a dilution factor of 1:6. % remaining HMGCR mRNA levels are displayed. 19PAT059649-WO-PCT

[0130] Figure 12: Dose response curve in Hepa-1-6 cells transfected with nine siRNAs (S1-S9) of Example 10 and with reporter plasmid TR030 in a 7-point dilution series starting at 6.7 nM with a dilution factor of 1:6. % remaining luciferase signal was normalized to renilla signal.

[0131] Figure 13: Dose response curve in Hepa-1-6 cells transfected with nine siRNAs (S1-S9) in Example 10 and with reporter plasmid TR029 in a 7-point dilution series starting at 6.7 nM with a dilution factor of 1:6. % remaining luciferase signal was normalized to renilla signal.

[0132] Figures 14A-14B: Liver HMGCR mRNA concentrations for male C57BL / 6 mice (n=3 or 4 per group) administered a single subcutaneous dose of PBS (white), No1 and No2 siRNAs at 3 mg / kg are shown in Figure 14A. No1 and No2 siRNAs share a same nucleotide sequence but have different chemical modifications. These results show durable HMGCR knockdown in mice, with No1 siRNA (with 2'-MOE clamp in the sense strand) performing better than No5 siRNA (without 2'-MOE clamp in the sense strand) at both timepoints evaluated (10 days or 42 days). Incorporation of guide strand into liver RISC over time is illustrated in Figure 14B. Male C57BL / 6 mice (n= 3 or 4 per group) were administered a single subcutaneous dose of PBS, No1 siRNA, or No5 siRNA at 3 mg / kg. Statistical significance was determined by ordinary one-way ANOVA and Sidak′s multiple comparisons test. Consistent with the liver HMGCR mRNA results, No1 siRNA showed the higher RISC- loading than No5 siRNA at both timepoints evaluated (10 days or 42 days).

[0133] Figures 15A-15B: Liver HMGCR mRNA concentrations for male C57BL / 6 mice (n=3 or 4 per group) administered a single subcutaneous dose of PBS (white), No1 to No4 siRNAs at 3 mg / kg are shown in Figure 15A. No1 to No4 siRNAs share a same nucleotide sequence but have different chemical modifications. These results show durable HMGCR knockdown in mice, and No1 and No2 (with MOE clamp and six or eight 3'-PS in the sense strand) performed similarly as No3 and No4 (with TNA clamp and six or eight 3'-PS in the sense strand) in this experiment. Incorporation of guide strand into liver RISC over time is illustrated in Figure 15B. Male C57BL / 6 mice (n= 3 or 4 per group) were administered a single subcutaneous dose of PBS, No1 to No4 siRNA at 3 mg / kg. Statistical significance was determined by ordinary one-way ANOVA and Sidak′s multiple comparisons test. Consistent with the liver HMGCR mRNA results, No1 to No4 siRNAs showed similar RISC-loading at 42 days. 20PAT059649-WO-PCT

[0134] Figure 16: Luciferase reporter assay results with Compounds 11-13 (Example 12) targeting HMGCR mRNA at position 115 in comparison to Compound 10 (targeting HMGCR mRNA at position 126).

[0135] Figure 17: Luciferase reporter assay results with Compounds 14-16 (Example 12) targeting HMGCR mRNA at position 2835 in comparison to Compound 10 (targeting HMGCR mRNA at position 126).

[0136] Figure 18: Percent reduction in plasma LDL-C concentrations in cynomolgus monkeys administered inclisiran and an HMGCR siRNA in Table 4 compared to the plasma LDL-C concentration administered inclisiran alone (inclisiran+PBS). DETAILED DESCRIPTION DEFINITIONS

[0137] Unless defined otherwise, all technical terms, scientific terms, abbreviations, chemical structures, and chemical formulae used herein have the same meaning as is commonly understood by one of ordinary skill in the art. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts. All patents, applications, published applications, and other publications referenced herein are incorporated by reference in their entirety unless stated otherwise.

[0138] All patents, applications, published applications, and other publications referenced herein are incorporated by reference in their entirety unless stated otherwise. Unless otherwise indicated, conventional methods of mass spectroscopy, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniques, and pharmacology are employed.

[0139] Furthermore, use of the term “including” as well as other forms, such as “include”, “includes,” and “included,” is not limiting. As used in this specification, whether in a transitional phrase or in the body of the claim, the terms “comprise(s)” and “comprising” are to be interpreted as having an open-ended meaning. That is, the terms are to be interpreted synonymously with the phrases “having at least” or “including at least.” When used in the context of a process, the term “comprising” means that the process includes at least the recited steps, but may include additional steps. When used in the context of a compound, composition, or device, the term “comprising” means that the compound, composition, or device includes at least the recited features or components, but may also include additional 21PAT059649-WO-PCT features or components. As used herein, the term "a,” "an,” "the” and similar terms used in the context of the present invention (especially in the context of the claims) are to be construed to cover both the singular and plural unless otherwise indicated herein or clearly contradicted by the context.

[0140] Unless otherwise indicated, all numbers, values, and / or expressions referring to nucleotide lengths, inhibition, activities, dosages, contents, and formulations used herein are to be understood as modified in all instances by the term “about” as such numbers are inherently approximations that are reflective of, among other things, the various uncertainties of measurement encountered in obtaining such values. Further, unless specifically stated or obvious from context, as used herein, the term “about” is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the “mean. “About” may be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from the context, all numerical values provided herein are modified by the term “about.”

[0141] The term “nucleic acid” means a compound containing at least two nucleotide monomers covalently linked together. Nucleic acids include polynucleotides and oligonucleotides, including double-stranded oligonucleotides and single-stranded oligonucleotides, and modified versions thereof.

[0142] The term “nucleotide” means a compound including a nucleoside and a phosphate group (or interchangeably, phosphodiester linkage) that are covalently attached at 5′ position or 3′ position of the pentofuranosyl sugar (e.g., ribose or deoxyribose). In certain aspects, the nucleotide is a ribonucleotide (RNA) having the ribose as the pentofuranosyl sugar. In certain aspects, a nucleotide is a deoxyribonucleotide (DNA) having the deoxyribose (2′- deoxyribose) as the pentofuranosyl sugar. Unless otherwise specifically indicated, when referring a “nucleotide” in a chain of nucleotides (e.g., oligonucleotides), e.g., X1to X21and X1′ to X23′, a nucleotide is meant by a nucleoside and a phosphate group (or phosphodiester linkage) that is covalently attached at 3′ position of the pentofuranosyl sugar (e.g., ribose or deoxyribose).

[0143] The term “nucleoside” means a monomer consisting of a nucleobase and a pentofuranosyl sugar (e.g., ribose or deoxyribose). A nucleoside including a ribose sugarring has to a structure a pharmaceutically acceptable salt thereof, and22PAT059649-WO-PCT a nucleotide including a deoxyribose sugar ring has a structu or a pharmaceutically acceptable salt, wherein in each structure, “ e.

[0144] The term “nucleobase” or “base,” as used herein,means the heterocyclic base moiety of a nucleoside or nucleotide. Non-limiting examples of nucleobases includes cytosine or a derivative thereof (e.g., cytosine analogue), guanine or a derivative thereof (e.g., guanine analogue), adenine or a derivative thereof (e.g., adenine analogue), thymine or a derivative thereof (e.g., thymine analogue), uracil or a derivative thereof (e.g., uracil analogue), hypoxanthine or a derivative thereof (e.g., hypoxanthine analogue), xanthine or a derivative thereof (e.g., xanthine analogue), 7-methylguanine or a derivative thereof (e.g., 7- methylguanine analogue), deaza-adenine or a derivative thereof (e.g., deaza-adenine analogue), deaza-guanine or a derivative thereof (e.g., deaza-guanine), deaza-hypoxanthine or a derivative thereof, 5,6-dihydrouracil or a derivative thereof (e.g., 5,6-dihydrouracil analogue), 5-methylcytosine or a derivative thereof (e.g., 5-methylcytosine analogue), or 5- hydroxymethylcytosine or a derivative thereof (e.g., 5-hydroxymethylcytosine analogue) moieties. In some embodiments, the nucleobase is adenine, guanine, hypoxanthine, xanthine, theobromine, caffeine, uric acid, or isoguanine, which may be optionally substituted or modified. In some embodiments, the nucleobase is,which may be optionally substituted or modified, wherein “ ” denotes the point ofattachment to a pentofuranosyl sugar ring (e.g., 1′ position).23PAT059649-WO-PCT

[0145] The term “phosphate,” or “phosphate group” as used herein a chemical species made of one phosphorus atom and four oxygen atom , or esters, salts, or acids thereof. In certain aspects, when the phosphate grou d between adjacent nucleosides in RNA or DNA strand and form a “backbone of the oligonucleotides, these terms “phosphate,” or “phosphate group” may be interchangeable used as “phosphate group,” “phosphate linkage,” “phosphodiester linkage,” or “linkage.” For example, the phosphate or phosphodiester linkage in the backbone of RNA or DNA may have the structures of or esters, salts (e.g., pharmaceutically acceptable salts), or acids (e.g.,”denotes the point of attachment to pentofuranosyl sugardjacent nucleosides. In certain aspects, a variant of aphosphate or phosphodiester linkage, e.g., phosphorothioate (PS) linkage, can replace a phosphate group (or phosphodiester linkage) in the backbone and connect two adjacent nucleosides. In certain aspects, a variant of a phosphate or phosphodiester linkage, e.g., phosphorothioate (PS) linkage or vinylphosphonate (VP) group, may be additionally attached at 3′ end or 5′ end of the oligonucleotides (e.g., RNA or DNA), e.g., 3′-OH or 5′-OH position of the terminal pentofuranosyl sugar (e.g., ribose or deoxyribose), so as to act as chemically or biologically functional group. In certain aspects, a variant of phosphate or phosphodiester linkage may also be referred as a phosphorus-derived internucleoside linkage that includes at least one phosphorus atom in the backbone.

[0146] Unless otherwise indicated herein, an unmodified RNA (or “ribonucleotide”) in a chain of nucleotides (e.g., mRNA, rRNA, or sense strand or antisense strand of siRNA) as disclosed refers to a structure of 24PAT059649-WO-PCT a pharmaceutically acceptable salt thereof. Likewise, an unmodified ucleotides”) in a chain of nucleotides (e.g., genomic DNA or cDNA) asdisclosed herein specifically refers to a structure of a pharmaceutically acceptable salt thereof. In each structure “Base” isn attachment point to the adjacent nucleotides.

[0147] Unless otherwise indicated herein, when an unmodified RNA is the first nucleotide from the 5′ end of an RNA chain (e.g., mRNA, or sense strand or antisense strand of siRNA), that nucleotide has a structur r a pharmaceutically acceptable salt thereof. Likewise, whenfirst nucleotide from the 5′ end of a DNA chain (e.g., genomic DNA or cDNA), that nucleotide has a structure of a pharmaceutically acceptable salt thereof. In each structure “Base” isn attachment point (5′ oxygen) to the adjacent nucleotides.

[0148] Alternatively, for example, the first nucleotide from the 5′ end of an RNA chain (e.g., mRNA, or sense strand or antisense strand of siRNA), that nucleotide has a structure of 25PAT059649-WO-PCT a pharmaceutically acceptable salt thereof and the first nucleotide A chain (e.g., genomic DNA or cDNA), that nucleotide has astructure a pharmaceutically acceptable salt thereof, when is an attachme adjacent nucleotides.

[0149] Unless otherwise indicated herein, when an unmodified RNA is the first nucleotide from the 3′ end of an RNA chain (e.g., mRNA, or sense strand or antisense strand of siRNA), that nucleotide has a structur r a pharmaceutically acceptable salt. Likewise, when an unmleotide from the 3′ end of a DNA chain (e.g., genomic DNA or cDNA), that nucleotide has a structure of a pharmaceutically acceptable salt thereof. In certain embodiments,A is the first nucleotide from the 3′ end of an RNA chain (e.g., mRNA, or sense strand or antisense strand of siRNA) that nucleotide does not include 3′ end phosphate group or phosphodiester linkage, for example, which has been removed during hydrolysis or synthesis, has a structur lly26PAT059649-WO-PCT acceptable salt. Likewise, when an unmodified DNA is the first nucleotide from the 3′ end of a DNA chain (e.g., genomic DNA or cDNA), that nucleotide does not include 3′ end phosphate group, for example, which has been removed during hydrolysis or synthesis, has a structure a pharmaceutically acceptable salt thereof. In each structure is an attachment point (e.g., phosphorus of thephosphate linkage) to the adjacent nucleotides.

[0150] A code “A”, “G”, “C”, or “U” presented in a sequence list as disclosed herein stand for a RNA nucleotide that contains adenine, guanine, cytosine, or uracil as a base, respectively. A code “dA”, “dG”, “dC” or “dT” presented in a sequence list as disclosed herein stand for a DNA nucleotide that contains adenine, guanine, cytosine, and thymine as a base, respectively. In some embodiments, the code “T” may be present in a RNA sequence then it may refer to a nucleotide (e.g. modified nucleotide) that thymine as a base.

[0151] The term “oligonucleotide” means a shorter length nucleic acid, e.g. of less than 100 nucleotides in length. Oligonucleotides may be single-stranded or double-stranded. In some embodiments, an oligonucleotide may include naturally occurring ribonucleotides, naturally occurring deoxyribonucleotides, and / or nucleotides having one or more modifications to a naturally occurring terminus, sugar, nucleobase, and / or internucleoside linkage. Non-limiting examples of oligonucleotides include double-stranded oligonucleotides (e.g., dsRNA), single-stranded oligonucleotides (e.g., single stranded RNA or ssRNA), antisense oligonucleotides (“ASO”), small interfering RNA (siRNA), microRNA mimics, short hairpin RNAs (shRNA), single-strand small interfering RNA (ssRNAi), RNaseH oligonucleotides, anti-microRNA oligonucleotides, steric blocking oligonucleotides, exon-skipping oligonucleotides, CRISPR guide RNAs, and aptamers. In certain aspects, the oligonucleotide is a dsRNA and each strand has a length less than 100 nucleotides (“nt”), less than 90 nt, less than 80 nt, less than 70 nt, less than 60 nt, less than 50 nt, less than 40 nt, less than 35 nt, less than 30 nt, less than 28 nt, less than 26 nt, less than 25 nt, less than 24 nt, less than 23 nt, less than 22 nt, less than 21 nt, less than 20 nt, less than 19 nt, less than 18 nt, less than 17 nt, less than 16 nt, or 15 nt.

[0152] The terms “iRNA”, “RNAi agent,” “iRNA agent,”, “RNA interference agent” as used interchangeably herein, refer to an agent that contains RNA as that term is defined herein, and which mediates the targeted cleavage of an RNA transcript (mRNA) via an RNA- 27PAT059649-WO-PCT induced silencing complex (RISC) pathway. An RNAi agent directs the sequence-specific degradation of mRNA through a process and thereafter inhibits expression of the gene encoded by the mRNA in a cell in vivo, e.g., in a subject (e.g., any vertebrate, mammal, or human).

[0153] The term “small interfering RNA” or “siRNA” means a double-stranded oligonucleotide (dsRNA) formed with two anti-parallel, and partially, substantially or fully complementary nucleic acid strands (e.g., a first strand and a second strand; or a “sense” strand and an “antisense” strand), which interferes with the expression of genes in a sequence-specific manner by facilitating mRNA degradation before translation through the RNA interference pathway. In some embodiments, depending on the context, the first strand can be a “guide” or antisense strand, and the second strand can be a “passenger” or sense strand. In some embodiments, depending on the context, the “first” strand can be a passenger or sense strand, and the “second” strand can be a guide or antisense. In certain aspects, an “RNAi agent” or “siRNA agent,” as used herein, refers a double-stranded RNA (dsRNA) with or without a ligand or other conjugate, and may be interchangeably used with a term “double stranded RNAi agent (dsRNAi agent),” or “dsRNA agent.” In certain aspect of the disclosure, the term “siRNA” can be used to describe a dsRNA with specific nucleotide sequences (unmodified or modified nucleotide sequences), without a ligand or other conjugate.

[0154] The term “antisense strand,” as used herein, refers an oligonucleotide (e.g., RNA) of an siRNA or a dsRNAi that is complementary (e.g., partially, substantially, or fully complementary) to the target mRNA and is incorporated into the RNA-induced silencing complex (RISC) to direct gene silencing in a sequence-specific manner through the RNA interference pathway. An antisense strand may also be referred to as the “guide strand.” In some embodiments, the antisense strand may have a length from 15-30 nt, 15-26 nt, 15-23 nt, 15-22 nt, 15-21 nt, 15-20 nt, 15-19 nt, 15-18 nt, 15-17 nt, 18-30 nt, 18-26 nt, 18-23 nt, 18-22 nt, 18-21 nt, 18-20 nt, 19-30 nt, 19-26 nt, 19-23 nt, 19-22 nt, 19-21 nt, 19-20 nt, 19 nt, 20-30 nt, 20-26 nt, 20-25 nt, 20-24 nt, 20-23 nt, 20-22 nt, 20-21 nt, 20 nt, 21-30 nt, 21-26 nt, 21-25 nt, 21-24 nt, 21-23 nt, 21-22 nt, 9 nt, 10 nt, 11 nt, 12 nt, 13 nt, 14 nt, 15 nt, 16 nt, 17 nt, 18 nt, 19 nt, 20 nt, 21 nt, 22 nt, 23 nt, 24 nt, 25 nt, 26 nt, 27 nt, 28 nt, 29 nt, 30 nt, 31 nt, 32 nt, 33 nt, 34 nt, 35 nt, or 36 nt.

[0155] The term “sense strand,” as used herein, refers an oligonucleotide that is complementary (e.g., partially, substantially, or fully complementary) to the antisense strand. 28PAT059649-WO-PCT The sense strand is typically degraded following incorporation of the antisense strand into RISC. The sense strand may also be referred to as the “passenger strand.” In some embodiments, the sense strand may have a length from 15-30 nt, 15-26 nt, 15-23 nt, 15-22 nt, 15-21 nt, 15-20 nt, 15-19 nt, 15-18 nt, 15-17 nt, 18-30 nt, 18-26 nt, 18-23 nt, 18-22 nt, 18-21 nt, 18-20 nt, 19-30 nt, 19-26 nt, 19-23 nt, 19-22 nt, 19-21 nt, 19-20 nt, 19 nt, 20-30 nt, 20-26 nt, 20-25 nt, 20-24 nt, 20-23 nt, 20-22 nt, 20-21 nt, 20 nt, 21-30 nt, 21-26 nt, 21-25 nt, 21-24 nt, 21-23 nt, 21-22 nt, 9 nt, 10 nt, 11 nt, 12 nt, 13 nt, 14 nt, 15 nt, 16 nt, 17 nt, 18 nt, 19 nt, 20 nt, 21 nt, 22 nt, 23 nt, 24 nt, 25 nt, 26 nt, 27 nt, 28 nt, 29 nt, 30 nt, 31 nt, 32 nt, 33 nt, 34 nt, 35 nt, or 36 nt.

[0156] The term “complementary” means that a nucleotide (e.g., RNA or DNA) or a sequence of nucleotides are capable of base pairing non-covalently via hydrogen bonding with another nucleotide or sequence of nucleotides. As described herein and commonly known in the art the complementary (matching) nucleotide of adenosine is thymidine or uridine and the complementary (matching) nucleotide of guanosine is cytidine. The complementarity of sequences may be partial, in which only some of the nucleic acids match according to base pairing, or complete, where all the nucleic acids match according to base pairing. For example, two sequences that are complementary to each other, may have a specified percentage of nucleotides that participate in nucleobase-pairing (i.e., about 50% complementarity, preferably 50%, 55%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater complementarity over a specified region). In some embodiments, two sequences are partially complementary when the percentage of nucleotides that participate in nucleobase-pairing is about 50%, about 55%, about 65%, about 70%, about 75%, or about 80%, or ranges from about 50% to about 80%. In some embodiments, two sequences are substantially complementary when the percentage of nucleotides that participate in nucleobase-pairing is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 92%, about 93%, about 94%, or about 95%, or ranges from about 80% to about 95%.

[0157] Examples of complementary (e.g., partially, substantially, or fully complementary) sequences are sense and antisense sequences, wherein the sense sequence contains complementary (e.g., partially, substantially, or fully complementary) nucleotides to the antisense sequence and thus forms the complement of the antisense sequence. In certain aspects, a sense strand and an antisense strand of a double-stranded oligonucleotide (e.g., 29PAT059649-WO-PCT double stranded RNA) are substantially or fully complementary over their entire lengths. In some embodiments, a sense strand and an antisense strand of dsRNA are substantially or fully complementary over the entire length of the double-stranded region of the siRNA, and one or both termini of either strand comprises single-stranded nucleotides.

[0158] Another examples of complementary (e.g., partially, substantially, or fully complementary) sequences are an antisense strand and its target mRNA sequence. In certain aspects, an antisense strand is substantially or fully complementary to its target mRNA. For example, the complementary (e.g., partially, substantially, or fully complementary) sequences may be between an antisense strand and a coding region of the target mRNA, or a non-coding sequence of the target mRNA. In certain aspects, an antisense strand is substantially, or fully complementary to its target mRNA to reduce or eliminate off-target profile for and to improve down-regulation of the target gene (e.g., gene of the target mRNA sequence).

[0159] The terms “identical” or percent “identity,” in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., at least 60% identity, or at least 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or within a range defined by any of two of the preceding values, identity over a specified region when compared and aligned for maximum correspondence over a comparison window or designated region) as measured using a BLAST or BLAST 2.0 sequence comparison algorithms with default parameters described below, or by manual alignment and visual inspection (see, e.g., NCBI web site or the like). This definition also refers to, or may be applied to, the complement of a test sequence. The definition also includes sequences that have deletions and / or additions, as well as those that have substitutions. As described below, the preferred algorithms can account for gaps, insertions and the like. Alignment for purposes of determining percent sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2 or Megalign (DNASTAR) software. Appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full-length of the sequences being compared can be determined by known methods.

[0160] As used herein, “target sequence” or “target gene” refer to a contiguous portion of the nucleotide sequence of an mRNA molecule formed during the transcription of a gene 30PAT059649-WO-PCT including mRNA that is a product of RNA processing of a primary transcription product. The target portion of the sequence will be at least long enough to serve as a substrate for RNAi- directed cleavage at or near that portion. For example, the target sequence will generally be from 9-36 nucleotides (“nt”) in length, e.g., 15-30 nt in length, including all sub-ranges therebetween. As non-limiting examples, the target sequence may have a length from 15-30 nt, 15-26 nt, 15-23 nt, 15-22 nt, 15-21 nt, 15-20 nt, 15-19 nt, 15-18 nt, 15-17 nt, 18-30 nt, 18- 26 nt, 18-23 nt, 18-22 nt, 18-21 nt, 18-20 nt, 19-30 nt, 19-26 nt, 19-23 nt, 19-22 nt, 19-21 nt, 19-20 nt, 19 nt, 20-30 nt, 20-26 nt, 20-25 nt, 20-24 nt, 20-23 nt, 20-22 nt, 20-21 nt, 20 nt, 21- 30 nt, 21-26 nt, 21-25 nt, 21-24 nt, 21-23 nt, 21-22 nt, 9 nt, 10 nt, 11 nt, 12 nt, 13 nt, 14 nt, 15 nt, 16 nt, 17 nt, 18 nt, 19 nt, 20 nt, 21 nt, 22 nt, 23 nt, 24 nt, 25 nt, 26 nt, 27 nt, 28 nt, 29 nt, 30 nt, 31 nt, 32 nt, 33 nt, 34 nt, 35 nt, or 36 nt.

[0161] The term “ligand,” as used herein, refers to a compound or moiety that can impose characteristics to provide additional properties, e.g., affinity or cell delivery efficiency, to an RNAi (e.g., dsRNAi) as described herein. The ligand may be coupled or conjugated directly to the RNAi (e.g., sense strand or antisense strand of dsRNA), or indirectly to the RNAi agent (e.g., sense strand or antisense strand of dsRNA) via an intervening linker (“linker”). When a ligand is conjugated or coupled indirectly to the RNAi (e.g., dsRNA) via a linker, the ligand may be formed of a core moiety (e.g., targeting moiety) that has specific function to provide affinity or efficacy and the linker that provides merely an optimal distance, e.g., between the core moiety and the RNAi agent (dsRNA). In certain aspects, the term “ligand” embraces the ligand in combination with the linker. Examples of ligands or targeting moieties thereof may include, but not be limited to, one or more selected from a synthetic or natural compound, a peptide, an antibody, a carbohydrate (e.g., sugar moiety), or an additional nucleic acid.

[0162] The term “modified nucleotide” means a nucleotide having one or more modifications relative to a naturally occurring nucleotide, e.g., RNA. The modified nucleotide may be selected over an unmodified form because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for other oligonucleotides or nucleic acid targets, increased stability in the presence of nucleases, and / or reduced immune stimulation. In certain aspects, the modification may be present in at least one of (i) an internucleoside linkage (“linkage”), (ii) a nucleobase, and (iii) a sugar moiety of the nucleotide. In certain aspects, the modification is present in the internucleoside linkage, e.g., by chemically modifying a phosphate (or phosphodiester) linkage or replacing a phosphate 31PAT059649-WO-PCT (or phosphodiester) linkage with other linking groups. In certain aspects, the modification is present in a sugar moiety, i.e., ribose ring, by substituting hydroxyl group on 2′ position of the ribose ring with other chemical group or by replacing a ring structure with otherheterocycloalkyl or cycloalkyl, glycol group having a structure , bicyclic orbridged ring on the ribose such as locked nucleic acid (LNA) hastructure of or the like. In certain aspects, the modification is present in a nucleobase (e.g., U) by chemical modification in a nucleobase by replacing the nucleobase withother moiety, for example, by replacing one naturally occurring nucleobase with another naturally occurring nucleobase. In certain aspects, a modified nucleotide may contain a modification in a sugar moiety and an unmodified phosphate (or phosphodiester) linkage. In certain aspects, a modified nucleotide may have a modification in a sugar moiety but with an unmodified nucleobase. In certain aspects, a modified nucleotide may have a modification in a sugar moiety and a nucleobase. In certain aspects, a modified nucleotide may have a modification in a sugar moiety and a phosphate (or phosphodiester) linkage. In certain aspects, a modified nucleotide may have a modification in a sugar moiety, a phosphate (or phosphodiester) linkage and a nucleobase. In certain aspects, a modified nucleotide may have an unmodified sugar moiety and an unmodified phosphate (or phosphodiester) linkage. In certain aspects, a modified nucleotide may have an unmodified sugar moiety and an unmodified nucleobase. In certain aspects, a modified nucleotide may have an unmodified sugar moiety and a modified nucleobase. In certain aspects, a modified nucleotide may have an unmodified sugar moiety and a modified phosphate (or phosphodiester) linkage. In certain aspects, a modified nucleotide may have a modified sugar moiety, a modified phosphate (or phosphodiester) linkage and a modified nucleobase.

[0163] The term “modified phosphate group,” or “modified phosphodiester linkage” as used herein refers to a chemical group in place of a phosphate group (or phosphodiester linkage) in a nucleotide as being attached to the 3′ end (3′ carbon) of the pentofuranosyl group.

[0164] The terms “identical” or percent “identity,” in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the 32PAT059649-WO-PCT same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., at least 60% identity, or at least 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or within a range defined by any of two of the preceding values, identity over a specified region when compared and aligned for maximum correspondence over a comparison window or designated region) as measured using a BLAST or BLAST 2.0 sequence comparison algorithms with default parameters described below, or by manual alignment and visual inspection (see, e.g., NCBI web site or the like). This definition also refers to, or may be applied to, the complement of a test sequence. The definition also includes sequences that have deletions and / or additions, as well as those that have substitutions. As described below, the preferred algorithms can account for gaps, insertions and the like. Alignment for purposes of determining percent sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2 or Megalign (DNASTAR) software. Appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full-length of the sequences being compared can be determined by known methods.

[0165] Throughout the disclosure, nucleotide positions or coordinates are relative to the beginning (5′ end) of the reference transcript.

[0166] The term “overhang” or “nucleotide overhang” herein refers to at least one unpaired nucleotide that protrudes from the end of at least one of the two strands of the duplex structure of an RNAi agent. In some embodiments, when a 3′-end of one strand extends beyond the 5′-end of the other strand, or vice versa, this forms a nucleotide overhang, e.g., the unpaired nucleotide(s) form the overhang.

[0167] “Blunt” or “blunt end” means that there are no unpaired nucleotides at that end of the double stranded RNAi agent, i.e., no nucleotide overhang. A “blunt ended” RNAi agent is a dsRNA that is double-stranded over its entire length, i.e., no nucleotide overhang at either end of the molecule.

[0168] A “mismatch” is defined herein as a difference between the base sequence (e.g., A instead of G) or length when two sequences are maximally aligned and compared. In certain aspects, the term “mismatch” means a nucleobase of a first oligonucleotide (e.g., a first strand) that is not capable of pairing with a nucleobase at a corresponding position of a second oligonucleotide (e.g., a second strand). 33PAT059649-WO-PCT

[0169] The term “non-end” herein refers to a position between the 3′ end and the 5′ end of the sense or antisense strand.

[0170] The term “3-hydroxy-3-methylglutaryl-CoA reductase,” as used herein and also interchangeably used with the term “HMGCR,” refers to a gene or protein (e.g., enzyme or reductase) thereof that converts 3-hydroxy-3-methylglutaryl coenzyme A (“HMG-CoA”) to mevalonate in cholesterol or isoprenoid synthesis. The term “HMGCR” also includes isoforms of proteins encoded by HMGCR mRNA sequences expressed in any vertebrate or mammals (e.g., human, mouse, rat, monkey, dog, cat, horse, pig, or cow).

[0171] In certain aspects, HMGCR may be identified with NCBI Gene ID, for example, human HMGCR Gene ID: 3156, mouse HMGCR Gene ID: 15357, rat HMGCR Gene ID: 25675, Rhesus monkey HMGCR Gene ID: 705479, dog HMGCR Gene ID: 479182, or cat HMGCR Gene ID: 101098922.

[0172] In certain aspects, HMGCR may be identified with mRNA transcript, for example, human HMGCR mRNA transcript (e.g., NM_000859.3; NM_001130996.2; and NM_001364187.1), mouse HMGCR mRNA transcript (e.g., NM_008255.2; NM_001360165.1; and NM_001360166.1), rat HMGCR mRNA transcript (e.g., NM_013134.2), Cynomolgus monkey HMGCR mRNA (e.g., XM_005557178.1), Rhesus monkey HMGCR mRNA (e.g., XM_001104607.4; and XM_002804417.3), dog HMGCR mRNA (e.g., XM_038471609.1; XM_038471611.1; and XM_038471610.1) and cat HMGCR mRNA (e.g., XM_003981075.6; and XM_019835641.3). In certain aspects, HMGCR may be identified with amino acid sequences, such as such as human HMGCR protein (e.g., NP_000850.1; NP_001124468.1; and NP_001351116.1), mouse HMGCR protein (e.g., NP_032281.2; NP_001347094.1; and NP_001347095.1), rat HMGCR protein (e.g., NP_037266.2), Rhesus monkey protein (e.g., XP_001104607.3; and XP_002804463.3), dog HMGCR protein (e.g., XP_038327537.1; XP_038327539.1; and XP_038327538.1), or cat HMGCR protein (e.g., XP_003981124.1 and XP_019691200.1). Examples of HMGCR gene, mRNA and proteins are not limited to the above list and may further include examples publicly available information from web database, for example, in GenBank, UniProt, Ensembl, Alliance and the like.

[0173] In certain aspects, HMGCR may include a fragment, variant, or mutant of the protein that may have the same or similar amino acid sequences (e.g., having about 80%, 85%, 90%, 95%, or 99% or greater of similarity or identity of amino acid sequences) with any one of the above listed HMGCR gene (mRNA) or protein sequences. In certain aspects, 34PAT059649-WO-PCT HMGCR may include a fragment, variant, or mutant of the protein having the same or in similar in vivo or in vitro enzymatic (e.g., reductase) activity, for example, having about 80%, 85%, 90%, 95%, or 99% or the native enzyme activity, to produce mevalonate from HMG-CoA.

[0174] The term “Compound” as used herein refers to a double stranded RNA (e.g., HMGCR dsRNA or dsRNAi agent) that is conjugated with a ligand or a delivery moiety, while a term “compound” denotation may refer to a substance, molecule or chemical entity that can be chemically defined and / or identifiable.

[0175] As defined herein, the term “inhibition”, “inhibit”, “inhibiting” and the like mean negatively affecting (e.g. decreasing) activity, expression or function relative to the activity, expression or function in the absence of an inhibitor. In certain aspects, inhibition can mean negatively affecting (e.g. decreasing) the concentration or levels of a biomolecule, such as a protein or mRNA, relative to the concentration or level of the biomolecule in the absence of an inhibitor. In certain aspects, inhibition includes, partially or totally, blocking stimulation, decreasing, preventing, or delaying activation; inactivating, desensitizing, or down-regulating signal transduction or enzymatic activity; or decreasing the amount of a biomolecule target (e.g., protein target or mRNA target). In certain aspects, inhibition refers to a reduction in the expression of a particular biomolecule target, such as a protein target (e.g., HMGCR protein) or an mRNA target (e.g., HMGCR mRNA). In certain aspects, inhibition refers to a reduction of amount of a target biomolecule (e.g., HMGCR protein or mRNA) resulting from a down-regulating protein expression (e.g. directly inhibiting translation or transcription). In certain aspects, inhibition refers to a reduction of activity of a target biomolecule (e.g., HMGCR protein or mRNA) from an indirect interaction (e.g., inhibiting or regulating other transcriptional or translational factors).

[0176] The term “inhibitor” also refers to a compound, composition, or substance capable of detectably negatively affecting (e.g. decreasing) activity, expression or function of a given protein or gene. For example, an inhibitor may decrease activity, expression or function by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater in comparison to a control in the absence of the inhibitor. Inhibitors include, for example, synthetic or biological molecules, such as oligonucleotides. In some embodiments, the inhibitors include RNAi agent, e.g., siRNA agent, dsRNAi agent, or dsRNA agent.

[0177] As used herein, the “level or degree of inhibiting or decreasing expression” of a given gene refers to the at least partial suppression of the expression of a target gene (e.g., 35PAT059649-WO-PCT HMGCR), as manifested by a reduction of the amount of the target gene mRNA (e.g., HMGCR mRNA) or protein (e.g., HMGCR) encoded by the target gene, which may be isolated from or detected in a group of cells (“a first cell”) in which a target gene is transcribed and which has or have been treated such that the expression of a target gene is inhibited, as compared to group of cells substantially identical to the first cell but without treated (“control cells” or “a second cell”).

[0178] In some embodiments, the level or expression of the target gene (e.g., HMGCR) can be measured by evaluation of mRNA (e.g., via Northern blots or PCR). The effect of an RNAi agent on the target gene (e.g., HMGCR) expression can be determined by measuring the gene transcription rates (e.g., via Northern blots; or reverse transcriptase polymerase chain reaction or real-time polymerase chain reaction). In some embodiments, the degree of inhibition can be calculated as the following equation: (mRNAin control cells) - (mRNA in treated cells)•100 % (mRNAin control cells)

[0179] Alternatively, the degree of inhibition may be given in terms of a reduction of a parameter that is functionally linked to target gene (e.g., HMGCR) expression, e.g., the amount of protein encoded by a target gene (e.g., HMGCR), alteration in expression of the protein whose expression is dependent on the target gene (e.g., HMGCR), alteration in an activity of the enzyme (e.g., HMGCR) encoded by the target gene (e.g., HMGCR). In some embodiments, the level or expression of the protein (e.g., HMGCR) from the target gene can be evaluated by measuring the expressed protein amount (e.g., Western blots). In some embodiments, the level or expression of the protein from the target gene can be measured by the enzymatic assay (e.g., kinetic assay) of the protein.

[0180] As used herein, the term “down-regulate” or “down-regulating” refers to any statistically significant decrease in a biological activity and / or expression of the target protein (e.g., HMGCR), including full blocking of the activity (i.e., complete inhibition) and / or expression. For example, “down-regulation” can refer to a decrease of at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% in target protein (e.g., HMGCR) level, activity and / or expression.

[0181] As used herein, the terms “salt” or “salts” refers to an acid addition or base addition salt of a compound of the present invention. “Salts” include in particular “pharmaceutical acceptable salts”. The term “pharmaceutically acceptable salts” refers to salts that retain the biological effectiveness and properties of the compounds of this invention 36PAT059649-WO-PCT and, which typically are not biologically or otherwise undesirable. In many cases, the compounds of the present invention are capable of forming acid and / or base salts by virtue of the presence of amino and / or carboxyl groups or groups similar thereto. When both a basic group and an acid group are present in the same molecule, the compounds of the present invention may also form internal salts, e.g., zwitterionic molecules. In certain aspects, pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids. Examples of the inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Examples of the organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid, and the like. In certain aspects, the pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Examples of the inorganic bases from which salts can be derived include, for example, ammonium salts and metals from columns I to XII of the periodic table, such as sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper; particularly suitable salts include ammonium, potassium, sodium, calcium and magnesium salts. Examples of the organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, such as organic amines include isopropylamine, benzathine, cholinate, diethanolamine, diethylamine, lysine, meglumine, piperazine and tromethamine.

[0182] In certain aspects, the term “pharmaceutically acceptable salt” as used herein may include the salts forms in acetate, ascorbate, adipate, aspartate, benzoate, besylate, bromide / hydrobromide, bicarbonate / carbonate, bisulfate / sulfate, camphorsulfonate, caprate, chloride / hydrochloride, chlortheophyllonate, citrate, ethandisulfonate, fumarate, gluceptate, gluconate, glucuronate, glutamate, glutarate, glycolate, hippurate, hydroiodide / iodide, isethionate, lactate, lactobionate, laurylsulfate, malate, maleate, malonate, mandelate, mesylate, methylsulphate, mucate, naphthoate, napsylate, nicotinate, nitrate, octadecanoate, oleate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, polygalacturonate, propionate, sebacate, stearate, succinate, sulfosalicylate, sulfate, tartrate, tosylate trifenatate, trifluoroacetate or xinafoate. 37PAT059649-WO-PCT

[0183] As used herein, the term "pharmaceutically acceptable carrier" refers to a substance useful in the preparation or use of a pharmaceutical composition and includes, for example, suitable diluents, solvents, dispersion media, surfactants, antioxidants, preservatives, isotonic agents, buffering agents, emulsifiers, absorption delaying agents, salts, drug stabilizers, binders, excipients, disintegration agents, lubricants, wetting agents, sweetening agents, flavoring agents, dyes, and combinations thereof, as would be known to those skilled in the art (see, for example, Remington The Science and Practice of Pharmacy, 22nd Ed. Pharmaceutical Press, 2013, pp.1049-1070).

[0184] As used herein, the term “treat,” “treating,” or “treatment” of any disease or disorder refers to alleviating or ameliorating the disease or disorder (i.e., slowing or arresting the development of the disease or at least one of the clinical symptoms thereof); or alleviating or ameliorating at least one physical parameter or biomarker associated with the disease or disorder, including those which may not be discernible to the patient. In some embodiments, treating does not include preventing.

[0185] As used herein, the term “prevent”, “preventing" or “prevention” of any disease or disorder refers to the prophylactic treatment of the disease or disorder; or delaying the onset or progression of the disease or disorder.

[0186] The term “therapy,” as used herein refers to an application of one or more specific procedures used for the amelioration of at least one indicator or a disease or condition. In certain aspects, the specific procedure is the administration of one or more pharmaceutical or therapeutic agents.

[0187] The term “associated” or “associated with” in the context of a substance or substance activity or function associated with a disease (e.g. a protein associated disease, or HMGCR associated disease) means that the disease is caused by (in whole or in part), or a symptom of the disease is caused by (in whole or in part) the substance or substance activity or function (e.g., HMGCR activity or function). Thus, as used herein, what is described as “being associated” with a disease, if a causative agent, could be a target for treatment of the disease.

[0188] The term “HMGCR-associated disorder or disease,” as used herein refers to a disorder or disease that is caused by, or associated with, HMGCR gene expression or HMGCR protein production. For example, HMGCR-associated disorder or disease (e.g. hyperlipidemia, hypercholesterolemia or ASCVD) may be treated with a HMGCR modulator (e.g., gene silencing agent or down regulator) or HMGCR inhibitor, in the instance where 38PAT059649-WO-PCT HMGCR activity or function (e.g., enzyme activity in cholesterol synthesis) controls key step in the cholesterol synthesis or metabolism.

[0189] As used herein, the term “hyperlipidemia” refers to any disorder, disease or condition characterized by abnormal elevation of levels of any or all lipids, such as cholesterol and triglycerides, and / or lipoproteins in the blood or a condition that can lead to abnormal elevation of levels of any or all lipids and / or lipoproteins in the blood. In one embodiment, the hyperlipidemia is hypertriglyceridemia. As used herein, the term “hypertriglyceridemia” refers to a condition in which triglyceride levels are elevated, often caused or exacerbated by uncontrolled hyperlipidemia mellitus, obesity, and sedentary habits, e.g., when triglycerides in blood are greater than 1000-2000 mg / dL. As used herein the term “hypercholesterolemia” refers to a form of hyperlipidemia (elevated levels of lipids in the blood) in which there are high levels of cholesterol in the serum of a subject, e.g., at least about 240 mg / dL of total cholesterol.

[0190] As used herein, the term “administering” means oral administration, administration as a suppository, topical contact, intravenous, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal or subcutaneous administration, or the implantation of a slow-release device, e.g., a mini-osmotic pump, to a subject. Administration is by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal) compatible with the preparation. Parenteral administration includes, e.g., intravenous, intramuscular, intra-arteriole, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, etc.

[0191] The term “combination,” embraces mixtures of first and second dsRNAi agents. The term “combination,” also embraces first and second dsRNAi agents, which are formulated separately. In some embodiments, the first dsRNAi agent and / or the second dsRNAi agent are administered together with one or more additional therapeutic agents. In certain embodiments, the first dsRNAi agent is administered at the same time, prior to, or after the administration of the second dsRNAi agent. In certain embodiments, the first dsRNAi agent and the second dsRNAi agent are administered together. In some embodiments, the first dsRNAi agent is formulated with one or more additional therapeutic agents, optionally in the same pharmaceutical composition as the first dsRNAi agent. In some embodiments, the second dsRNAi agent is formulated with one or more additional therapeutic agents, optionally in the same pharmaceutical composition as the second dsRNAi 39PAT059649-WO-PCT agent. In some embodiments, the first dsRNAi agent and the second dsRNAi agent are formulated with one or more additional therapeutic agents, optionally in the same pharmaceutical composition as the first dsRNAi agent and the second dsRNAi agent. In some embodiments, the first dsRNAi agent (e.g., an HMGCR dsRNAi agent described herein) and the second dsRNAi agent (e.g., inclisiran) are formulated as a fixed dose combination (“FDC”).

[0192] Co-administration includes administering one active agent (e.g., RNAi agent) within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours of a second active agent (e.g. anticancer or antitumor agents). Also contemplated herein, are embodiments, where co-administration includes administering one active agent (e.g., dsRNAi agent or a therapeutic agent) within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours of a second active agent. Co-administration includes administering two active agents simultaneously, approximately simultaneously (e.g., within about 1, 5, 10, 15, 20, or 30 minutes of each other), or sequentially in any order. In some embodiments, co-administration can be accomplished by co-formulation, i.e., preparing a single pharmaceutical composition including both active agents. In some embodiments, the active agents can be formulated separately.

[0193] In some embodiments, the first dsRNAi agent and the second dsRNAi agent are co-administered. Co-administration includes administering the first dsRNAi agent within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours of the second dsRNAi agent. Co-administration includes administering the first dsRNAi agent and the second dsRNAi agent simultaneously, approximately simultaneously (e.g., within about 1, 5, 10, 15, 20, or 30 minutes of each other), or sequentially in any order. In some embodiments, co-administration can be accomplished by co-formulation, i.e., preparing a single pharmaceutical composition including the first dsRNAi agent and the second dsRNAi agent. In some embodiments, the first dsRNAi agent and the second dsRNAi agent are formulated separately.

[0194] The terms "subject" and “patient” as used herein are used interchangeably. The term subject includes a human or non-human animal, preferably a vertebrate, and more preferably a mammal. In certain aspects, the subject is a human. In certain aspects, the subject is a human patient.

[0195] As used herein, a subject is “in need of” a treatment if such subject would benefit biologically, medically or in quality of life from such treatment.

[0196] The term "a therapeutically effective amount" of a compound (e.g., siRNA) as disclosed herein refers to an amount of the compound that will elicit the biological or medical 40PAT059649-WO-PCT response of a subject, for example, reduction or inhibition of an enzyme or a protein activity, or ameliorate symptoms, alleviate conditions, slow or delay disease progression, or prevent a disease, etc. In certain aspects, the term “a therapeutically effective amount” refers to the amount of the compound (e.g., siRNA) of the disclosure that, when administered to a subject, is effective to (1) at least partially alleviate, prevent and / or ameliorate a condition, or a disorder or a disease (i) mediated by the target gene (e.g., HMGCR), or (ii) associated with its activity, or (iii) characterized by activity (normal or abnormal) of the protein encoded by the target gene (e.g., HMGCR); or (2) reduce or inhibit the activity of the protein encoded by the target gene (e.g., HMGCR); or (3) reduce or inhibit the expression of the target gene (e.g., HMGCR). In certain aspects, the term “a therapeutically effective amount” refers to the amount of the compound that, when administered to a cell, or a tissue, or a non-cellular biological material, or a medium, is effective to at least partially reducing or inhibiting the activity of the protein encoded by the target gene (e.g., HMGCR); or at least partially reducing or inhibiting the expression of the protein (e.g., HMGCR) encoded by the target gene. The meaning of the term “a therapeutically effective amount” as illustrated in the above embodiment for the target gene expression also applies by the same means to any other relevant proteins / peptides / enzymes (e.g., HMGCR or other proteins relevant to cholesterol biosynthesis).

[0197] For any compound described herein, the therapeutically effective amount can be initially determined from cell culture assays. Target concentrations will be those concentrations of active compound(s) that are capable of achieving the methods described herein, as measured using the methods described herein or known in the art. Therapeutically effective amounts for use in humans can also be determined from animal models. For example, a dose for humans can be formulated to achieve a concentration that has been found to be effective in animals. The dosage in humans can be adjusted by monitoring compounds effectiveness and adjusting the dosage upwards or downwards, as described above. Adjusting the dose to achieve maximal efficacy in humans based on the methods established well within the capabilities of the ordinarily skilled artisan (e.g., physician or medical expert). An example of an “therapeutically effective amount” is an amount sufficient to contribute to the treatment, prevention, or reduction of a symptom or symptoms of a disease. For example, for the given parameter (e.g., biomarker), a therapeutically effective amount will show an increase or decrease of at least 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90%, or at least 100%. Therapeutic efficacy can also be expressed as “-fold” increase or decrease. 41PAT059649-WO-PCT For example, a therapeutically effective amount can have at least a 1.2-fold, 1.5-fold, 2-fold, 5-fold, or more effect over a control.

[0198] The term “control” or “control experiment” is used in accordance with its plain ordinary meaning and refers to an experiment in which the subjects or reagents of the experiment are treated as in a parallel experiment except for omission of a procedure, reagent, or variable of the experiment. Typically, a control is used as a standard of comparison in evaluating experimental effects. In some embodiments, a control is the measurement of the expression of a protein or mRNA (e.g., HMGCR) in the absence of RNAi agents as described herein.

[0199] The term “muscle side effect,” “muscle adverse effect,” or “ muscle-related side effect” as used herein refers to a symptom or negative side effect, such as pain (e.g., ranging from mild discomfort to serious pain), weakness, soreness, tiredness, damage (e.g., rhabdomyolysis), or spasms, caused in or near muscles or muscle tissues. In some embodiments, the muscle side effect may be a drug-induced myopathies, for example, caused by taking a medicine (e.g., statin).

[0200] Unless defined otherwise, the chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts.

[0201] The term “alkyl,” by itself or as part of another substituent, means, unless otherwise stated, a straight (i.e., unbranched) or branched carbon chain (or carbon), or combination thereof, which is fully saturated (i.e., molecule by only single bonds) and include mono-, di- and multivalent radicals. As used herein, the alkyl is an uncyclized chain. The alkyl may include a designated number of carbons (e.g., C1-C10means one to ten carbons). Examples of alkyl include, but are not limited to, groups such as C1-30 alkyl, C1-25 alkyl, C1-20alkyl, C1-15alkyl, C1-12alkyl, C1-10alkyl, C1-8alkyl, C1-6alkyl, C1-4alkyl, or C1-3alkyl. For example, C1-6 alkyl include, but are not limited to, methyl, ethyl, n-propyl, 1- methylethyl (iso-propyl), n-butyl, n-pentyl and 1,1-dimethylethyl (t-butyl), and their isomers.

[0202] A term “alkylene,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkyl, as exemplified, but not limited by, - CH2CH2CH2CH2-.

[0203] As used herein, the term "alkenyl," by itself or as part of another substituent, means, unless otherwise stated, a straight (i.e., unbranched) or branched carbon chain (or carbon), or combination thereof, which is mono- or polyunsaturated (i.e., molecule including 42PAT059649-WO-PCT at least one double bond) and include mono-, di- and multivalent radicals. As used herein, the alkenyl is an uncyclized chain. Like the alkyl, the alkenyl may include a designated number of carbons (e.g., C1-C10 means one to ten carbons). Examples of alkenyl include, but are not limited to, groups such as C1-30alkenyl, C1-25alkenyl, C1-20alkenyl, C1-15alkenyl, C1-12alkenyl, C1-10 alkenyl, C1-8 alkenyl, C1-6 alkenyl, C1-4 alkenyl, or C1-3 alkenyl. For example, C2-6alkenyl include, but are not limited to, ethenyl (vinyl), prop-1-enyl, but-1-enyl, pent-1- enyl, pent-4-enyl and penta-1,4-dienyl, and their isomers. A term “alkenylene,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkenyl, as exemplified, but not limited by, -CH=CHCH2CH2-.

[0204] As used herein, the term " alkynyl," by itself or as part of another substituent, means, unless otherwise stated, a straight (i.e., unbranched) or branched carbon chain (or carbon), or combination thereof, which is mono- or polyunsaturated (i.e., molecule including at least one triple bond) and include mono-, di- and multivalent radicals. As used herein, the alkynyl is an uncyclized chain. Like the alkyl, the alkynyl may include a designated number of carbons (e.g., C1-C10means one to ten carbons). Examples of alkynyl include, but are not limited to, groups such as C1-30 alkynyl, C1-25 alkynyl, C1-20 alkynyl, C1-15 alkynyl, C1-12 alkynyl, C1-10alkynyl, C1-8alkynyl, C1-6alkynyl, C1-4alkynyl, or C1-3alkynyl. For example, C2-6 alkynyl include, but are not limited to, alkynyl, and their isomers. A term “alkynyl,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from an alkenyl, as exemplified, but not limited by, -CCH2CH2-.

[0205] As used herein, the term “alkoxy” refers to a radical of the formula -ORawhere Rais an alkyl (e.g., C1-30 alkyl, C1-25 alkyl, C1-20 alkyl, C1-15 alkyl, C1-12 alkyl, C1-10 alkyl, C1-8 alkyl, C1-6alkyl, C1-4alkyl, or C1-3alkyl) radical as generally defined above. For example, C1-6 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, pentoxy, and hexoxy.

[0206] As used herein, the term " alkoxyalkyl " refers to a radical of the formula -Ra-O- Rbwhere each Raand Rbis independently an alkyl (e.g., C1-30alkyl, C1-25alkyl, C1-20alkyl, C1-15 alkyl, C1-12 alkyl, C1-10 alkyl, C1-8 alkyl, C1-6 alkyl, C1-4 alkyl, or C1-3 alkyl) radical as defined above and oxygen atom may be bonded to any carbon atom in either alkyl radical. For example, C1-6alkoxy C1-6alkyl include, but are not limited to, methoxy-methyl, methoxy- ethyl, ethoxy-ethyl, 1-ethoxy-propyl and 2-methoxy-butyl. 43PAT059649-WO-PCT

[0207] As used herein, the term “alkylcarbonyl” refers to a radical of the formula - C(=O)-Rawhere Rais an alkyl (e.g., C1-30alkyl, C1-25alkyl, C1-20alkyl, C1-15alkyl, C1-12alkyl, C1-10 alkyl, C1-8 alkyl, C1-6 alkyl, C1-4 alkyl, or C1-3 alkyl) radical as defined above.

[0208] As used herein, the term "alkyl-carbonyl alkyl” refers to a radical of the formula - Ra-C(=O)-Rbwhere each Raand Rbis independently an alkyl (e.g., C1-30 alkyl, C1-25 alkyl, C1-20alkyl, C1-15alkyl, C1-12alkyl, C1-10alkyl, C1-8alkyl, C1-6alkyl, C1-4alkyl, or C1-3alkyl) radical as defined above. The carbon atom of the carbonyl group may be bonded to any carbon atom in either alkyl radical.

[0209] As used herein, the term "alkylaminocarbonyl" refers to a radical of the formula - C(=O)-NH-Rawhere Rais an alkyl (e.g., C1-30alkyl, C1-25alkyl, C1-20alkyl, C1-15alkyl, C1-12alkyl, C1-10 alkyl, C1-8 alkyl, C1-6 alkyl, C1-4 alkyl, or C1-3 alkyl) as defined above.

[0210] As used herein, the term "alkoxycarbonyl” refers to a radical of the formula - C(=O)-O-Rawhere Rais an alkyl (e.g., C1-30 alkyl, C1-25 alkyl, C1-20 alkyl, C1-15 alkyl, C1-12 alkyl, C1-10 alkyl, C1-8 alkyl, C1-6 alkyl, C1-4 alkyl, or C1-3 alkyl) radical as defined above.

[0211] As used herein, the term “alkoxycarbonyl alkyl” refers to a radical of the formula -Ra-C(=O)-O-Rbwhere each Raand Rbis independently an alkyl (e.g., C1-30 alkyl, C1-25 alkyl, C1-20alkyl, C1-15alkyl, C1-12alkyl, C1-10alkyl, C1-8alkyl, C1-6alkyl, C1-4alkyl, or C1-3alkyl) radical as defined above.

[0212] As used herein, the term "haloalkyl" refers to an alkyl (e.g., C1-30alkyl, C1-25alkyl, C1-20 alkyl, C1-15 alkyl, C1-12 alkyl, C1-10 alkyl, C1-8 alkyl, C1-6 alkyl, C1-4 alkyl, or C1-3 alkyl) radical, as defined above, substituted by one or more halo radicals, as defined above. Examples of halogenC1-6alkyl include, but are not limited to, trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,3-dibromopropan-2-yl, 3-bromo-2- fluoropropyl and 1,4,4-trifluorobutan-2-yl.

[0213] As used herein, the term "hydroxyalkyl” refers to an alkyl (e.g., C1-30alkyl, C1-25alkyl, C1-20 alkyl, C1-15 alkyl, C1-12 alkyl, C1-10 alkyl, C1-8 alkyl, C1-6 alkyl, C1-4 alkyl, or C1-3 alkyl) radical as defined above, wherein one of the hydrogen atoms of the alkyl radical is replaced by OH. Examples of hydroxyC1-6 alkyl include, but are not limited to, hydroxy- methyl, 2-hydroxy-ethyl, 2-hydroxy-propyl, 3-hydroxy-propyl and 5-hydroxy-pentyl.

[0214] As used herein, the term “aminoalkyl” refers to an alkyl (e.g., C1-30 alkyl, C1-25 alkyl, C1-20alkyl, C1-15alkyl, C1-12alkyl, C1-10alkyl, C1-8alkyl, C1-6alkyl, C1-4alkyl, or C1-3alkyl) radical as defined above, wherein one of the hydrogen atoms of the C1-6alkyl group is replaced by a primary amino group. Examples of amino C1-6 alkyl include, but are not 44PAT059649-WO-PCT limited to, amino-methyl, 2-amino-ethyl, 2-amino-propyl, 3-amino-propyl, 3-amino-pentyl and 5-amino-pentyl.

[0215] As used herein, the term “alkylamino” refers to a radical of the formula -NH-Rawhere Rais an alkyl (e.g., C1-30alkyl, C1-25alkyl, C1-20alkyl, C1-15alkyl, C1-12alkyl, C1-10alkyl, C1-8 alkyl, C1-6 alkyl, C1-4 alkyl, or C1-3 alkyl) radical as defined above.

[0216] The term “heteroalkyl,” by itself or in combination with another term, means, unless otherwise stated, a stable straight or branched chain, or combination thereof, which is fully saturated (i.e., molecule by only single bonds) and include mono-, di- and multivalent radicals, including at least one carbon atom and at least one heteroatom (e.g., O, N, S, Si, or P), and wherein the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) (e.g., O, N, S, Si, or P) may be placed at any interior position of the heteroalkyl group or at the position at which the alkyl group is attached to the remainder of the molecule. Heteroalkyl is an uncyclized chain. The heteroalkyl may include a designated number of carbons and heteroatoms (e.g., “2 to 10 membered heteroalkyl” means two to 10 atoms including carbons and heteroatoms).

[0217] Similarly, the term “heteroalkylene,” by itself or as part of another substituent, means, unless otherwise stated, a divalent radical derived from heteroalkyl, as exemplified, but not limited by, -CH2-CH2-S-CH2-CH2- and -CH2-S-CH2-CH2-NH-CH2-. For heteroalkylene groups, heteroatoms can also occupy either or both of the chain termini (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, and the like).

[0218] As used herein, the term "heteroalkenyl," by itself or as part of another substituent, means, unless otherwise stated, a straight (i.e., unbranched) or branched carbon chain (or carbon), or combination thereof, which is mono- or polyunsaturated (i.e., molecule including at least one double bond between carbon and carbon) and include mono-, di- and multivalent radicals. As used herein, the alkenyl is an uncyclized chain. Like the alkenyl, the heteroalkenyl may include a designated number of carbons and heteroatoms (e.g., “2 to 10 membered heteroalkenyl” means two to 10 atoms including carbons and heteroatoms).

[0219] As used herein, the term " heteroalkynyl," by itself or as part of another substituent, means, unless otherwise stated, a straight (i.e., unbranched) or branched carbon chain (or carbon), or combination thereof, which is mono- or polyunsaturated (i.e., molecule including at least one triple bond between carbon and carbon) and include mono-, di- and multivalent radicals. As used herein, the alkynyl is an uncyclized chain. The heteroalkynyl 45PAT059649-WO-PCT may include a designated number of carbons and heteroatoms (e.g., “2 to 10 membered heteroalkynyl” means two to 10 atoms including carbons and heteroatoms).

[0220] For alkylene and heteroalkylene linking groups, no orientation of the linking group is implied by the direction in which the formula of the linking group is written. For example, the formula -C(O)2R′- represents both -C(O)2R′- and -R′C(O)2-.

[0221] A “cycloalkylene” and a “heterocycloalkylene,” alone or as part of another substituent, means a divalent radical derived from a cycloalkyl and heterocycloalkyl, respectively. The terms “cycloalkyl” and “heterocycloalkyl,” by themselves or in combination with other terms, mean, unless otherwise stated, cyclic versions of “alkyl” and “heteroalkyl,” respectively. Cycloalkyl and heterocycloalkyl are not aromatic. Additionally, for heterocycloalkyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples of heterocycloalkyl include, but are not limited to, 1- (1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3- morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, and the like. A “cycloalkylene” and a “heterocycloalkylene,” alone or as part of another substituent, means a divalent radical derived from a cycloalkyl and heterocycloalkyl, respectively.

[0222] The term “aryl” means, unless otherwise stated, a polyunsaturated, aromatic, hydrocarbon substituent, which can be a single ring or multiple rings (preferably from 1 to 3 rings) that are fused together (i.e., a fused ring aryl) or linked covalently. A fused ring aryl refers to multiple rings fused together wherein at least one of the fused rings is an aryl ring. The term “heteroaryl” refers to aryl groups (or rings) that contain at least one heteroatom such as N, O, or S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized. Thus, the term “heteroaryl” includes fused ring heteroaryl groups (i.e., multiple rings fused together wherein at least one of the fused rings is a heteroaromatic ring). A 5,6-fused ring heteroarylene refers to two rings fused together, wherein one ring has 5 members and the other ring has 6 members, and wherein at least one ring is a heteroaryl ring. Likewise, a 6,6-fused ring heteroarylene refers to two rings fused together, wherein one ring has 6 members and the other ring has 6 members, and wherein at least one ring is a heteroaryl ring. And a 6,5-fused ring heteroarylene refers to two rings fused together, wherein one ring has 6 members and the other ring has 5 members, and wherein at 46PAT059649-WO-PCT least one ring is a heteroaryl ring. A heteroaryl group can be attached to the remainder of the molecule through a carbon or heteroatom. Non-limiting examples of aryl and heteroaryl groups include phenyl, naphthyl, pyrrolyl, pyrazolyl, pyridazinyl, triazinyl, pyrimidinyl, imidazolyl, pyrazinyl, purinyl, oxazolyl, isoxazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrimidyl, benzothiazolyl, benzooxazoyl benzimidazolyl, benzofuran, isobenzofuranyl, indolyl, isoindolyl, benzothiophenyl, isoquinolyl, quinoxalinyl, quinolyl, 1-naphthyl, 2- naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4- imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3- thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3- quinolyl, and 6-quinolyl. Substituents for each of the above noted aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below. An “arylene” and a “heteroarylene,” alone or as part of another substituent, mean a divalent radical derived from an aryl and heteroaryl, respectively.

[0223] The terms “halo” or “halogen,” by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as “haloalkyl” are meant to include monohaloalkyl and polyhaloalkyl. For example, the term “halo(C1-C4)alkyl” includes, but is not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0224] The symbol “ ” denotes the point of attachment of a chemical moiety to theremainder of a molecule or chemical formula.

[0225] The term “oxo,” as used herein, means an oxygen that is double-bonded to a carbon atom.

[0226] Each of the above terms (e.g., “alkyl,” “heteroalkyl,” “cycloalkyl,” “heterocycloalkyl,” “aryl,” and “heteroaryl”) includes both substituted and unsubstituted forms of the indicated radical. Substituents for the alkyl and heteroalkyl radicals (including those groups often referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) can be one or more of a variety of groups selected from, but not limited to, —OR′, ═O, ═NR′, ═N—OR′, —NR′R″, —SR′, -halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO2R′, —CONR′R″, — OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)2R′, —NR— 47PAT059649-WO-PCT C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)2R′, —S(O)2NR′R″, — NRSO2R′, —NR′NR″R′″, —ONR′R″, —NR′C(O)NR″NR′″R″″, —CN, —NO2, — NR′SO2R″, —NR′C(O)R″, —NR′C(O)—OR″, —NR′OR″, in a number ranging from zero to (2m′+1), where m′ is the total number of carbon atoms in such radical. R, R′, R″, R′″, and R″″ each preferably independently refer to hydrogen, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl (e.g., aryl substituted with 1-3 halogens), substituted or unsubstituted heteroaryl, substituted or unsubstituted alkyl, alkoxy, or thioalkoxy groups, or arylalkyl groups. When a compound described herein includes more than one R group, for example, each of the R groups is independently selected as are each R′, R″, R′″, and R″″ group when more than one of these groups is present. When R′ and R″ are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 4-, 5-, 6-, or 7- membered ring. For example, —NR′R″ includes, but is not limited to, 1-pyrrolidinyl and 4- morpholinyl. From the above discussion of substituents, one of skill in the art will understand that the term “alkyl” is meant to include groups including carbon atoms bound to groups other than hydrogen groups, such as haloalkyl (e.g., -CF3 and -CH2CF3) and acyl (e.g., - C(O)CH3, —C(O)CF3, —C(O)CH2OCH3, and the like).

[0227] Certain compounds provided herein possess asymmetric carbon atoms (optical or chiral centers) or double bonds; the enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino acids, and individual isomers are encompassed within the scope of the present disclosure. The compounds of provided herein do not include those that are known in art to be too unstable to synthesize and / or isolate. Compounds provided herein include those in racemic and optically pure forms. Optically active (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When the compounds described herein contain olefinic bonds (vinyl group) and unless specified otherwise, it is intended that the compounds include both (E) and (Z) geometric isomers.

[0228] As used herein, the term “isomers” refers to compounds having the same number and kind of atoms, and hence the same molecular weight, but differing in respect to the structural arrangement or configuration of the atoms. 48PAT059649-WO-PCT RNAi AGENTS

[0229] In an aspect, the disclosure provides an RNAi agent including a double stranded RNA (dsRNA). In an aspect, also provided is a dsRNA interference (dsRNAi) agent that includes a dsRNA consisting of (i) a sense strand and (ii) an antisense strand, and a ligand attached to at least one of the sense strand and the antisense strand.

[0230] A dsRNA is a complex of ribonucleic acid (RNA) molecules formed in a duplex structure. In certain aspects, the dsRNA may be a short interfering RNA (siRNA) that has 10 to 30, or particularly 15-25 nucleotides in each RNA molecule, respectively, “passenger strand” and “guide strand”, and can be incorporated into an RNA-induced silencing complex (RISC). The siRNA is dissociated or unwounded in the RISC, and the passenger strand is degraded while the guide strand remains in the RISC pathway. The guide strand can subsequently bind to a mRNA molecule that includes a complementary sequence to the guide strand and induce or initiate cleavage or degradation of the mRNA molecule. In certain aspects, the mRNA encodes a target gene (e.g., mRNA transcript of a target gene) such that expression of the target gene is suppressed or inhibited through a post-transcriptional gene- silencing (“RNA silencing”). A guide RNA molecule has a complementary sequence to a target mRNA sequence and has anti-parallel orientation to the target gene, so it is interchangeably referred to as an antisense strand. A passenger RNA molecule forming a duplex with the guide RNA and having a complementary sequence to the guide strand (antisense strand) has the same orientation with the target mRNA sequence, so it is interchangeably referred to as an antisense strand. HMGCR siRNA (double stranded RNA)

[0231] In an aspect, the disclosure provides a dsRNA interference (dsRNAi) agent that is capable of interacting or recruiting a target mRNA sequence, e.g., HMGCR target mRNA sequence, in the RISC thereby cleaving the target mRNA. The dsRNAi agent can silence HMGCR gene, e.g., by inhibiting, downregulating, or suppressing the expression of HMGCR gene. Gene-silencing (e.g., inhibiting, downregulating, or suppressing of the gene) may be assessed by a decrease in an absolute or relative level of one or more variables that are associated with HMGCR expression compared with a control level. The control level may be any type obtained from, e.g., a pre-dose baseline level, or a level determined from a similar subject, cell, or untreated or treated subject with inactive agents (e.g., PBS buffer). In some embodiments, the level of silencing the HMGCR may be demonstrated by a reduction of the amount of a total HMGCR mRNA in a cell. In some embodiments, the level of silencing the 49PAT059649-WO-PCT HMGCR may be demonstrated by a reduction of the amount of a total HMGCR protein in a cell.

[0232] In an aspect, the dsRNAi agent as described herein can inhibit expression of the HMGCR gene (e.g., human HMGCR) by at least about 10%, about 15%, about 20%, about 25%, about 30 %, about 40%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is inhibited by at least about 10%, about 15%, about 20%, about 25%, about 30 %, about 40%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is inhibited by at least about 10% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is inhibited by at least about 20% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is inhibited by at least about 30% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is inhibited by at least about 40% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is inhibited by at least about 50% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is inhibited by at least about 60% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is inhibited by at least about 70% based on the expression level of the HMGCR gene in untreated cell or subject.

[0233] In an aspect, the dsRNAi agent as described herein can decrease expression of the HMGCR gene (e.g., human HMGCR) by at least about 10%, about 15%, about 20%, about 25%, about 30 %, about 40%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is decreased by at least about 10%, about 15%, about 20%, about 25%, about 30 %, about 40%, about 50%, about 55%, about 60%, about 65%, 50PAT059649-WO-PCT about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is decreased by at least about 10% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is decreased by at least about 20% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is decreased by at least about 30% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is decreased by at least about 40% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is decreased by at least about 50% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is decreased by at least about 60% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is decreased by at least about 70% based on the expression level of the HMGCR gene in untreated cell or subject.

[0234] In an aspect, the dsRNAi agent as described herein can suppress expression of the HMGCR gene (e.g., human HMGCR) by at least about 10%, about 15%, about 20%, about 25%, about 30 %, about 40%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is suppressed by at least about 10%, about 15%, about 20%, about 25%, about 30 %, about 40%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is suppressed by at least about 10% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is suppressed by at least about 20% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is suppressed by at least about 30% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., 51PAT059649-WO-PCT human HMGCR) is suppressed by at least about 40% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is suppressed by at least about 50% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is suppressed by at least about 60% based on the expression level of the HMGCR gene in untreated cell or subject. In some embodiments, expression of the HMGCR gene (e.g., human HMGCR) is suppressed by at least about 70% based on the expression level of the HMGCR gene in untreated cell or subject.

[0235] In some embodiments, inhibition of the expression of the HMGCR gene may be manifested by a reduction of the amount of mRNA expressed in a first cell or a first group of cells obtained from a subject that has been treated, e.g., by contacting the cell or by administering the dsRNAi agent as described herein, as compared to a second cell or a second group of cells obtained from a subject that has not been treated but is identical to the first cell or the first group of cells. For example, the level of gene-silencing (e.g., inhibiting, downregulating, or suppressing of the gene) of the HMGCR (e.g., human HMGCR) may be presented as a percentage of remaining mRNA in the treated cells (first cell or group of cells) compared to the mRNA amount in the control (untreated) cells, as shown in the following equation: (mRNA in control cells) - (mRNA in treated cells)•100 % (mRNA in control cells) .

[0236] In some embodiments, the level of gene-silencing (e.g., inhibiting, downregulating, or suppressing of the gene) of the HMGCR (e.g., human HMGCR) may be assessed by measuring a parameter or biomarker, e.g., human HMGCR protein level, in a biological sample (e.g., e.g., a blood, serum or liver tissue obtained from a subject), which may be treated or untreated. Conventional analytical methods as known in the art such as electrophoresis (e.g., SDS or capillary electrophoresis), chromatography (e.g., high performance liquid chromatography (HPLC)), spectroscopy, western blotting, enzyme- linked immunosorbent assays (ELISAs), immunofluorescent assays, electrochemiluminescence assays, and the like can be used without limitation, but examples are not limited thereto. In some embodiments, reduced level of gene-silencing (e.g., inhibiting, downregulating, or suppressing of the gene) of the HMGCR (e.g., human HMGCR) may be observed or assessed by in a liver (tissue) biopsy of the treated subject. 52PAT059649-WO-PCT

[0237] In certain aspects, the dsRNAi agent is a free acid. In certain aspects, the dsRNAi agent is in a salt form (e.g., a pharmaceutically acceptable salt form. It will be understood that references to dsRNAi agent are meant to also include the pharmaceutically acceptable salts of the dsRNAi agent. If the dsRNAi agent has, for example, at least one basic center, they can form acid addition salts. Corresponding acid addition salts can also be formed having, if desired, an additionally present basic center. Active substances having an acid group, e.g., COOH, can form salts with bases. The dsRNAi agent or pharmaceutically acceptable salts thereof may also be used in form of a hydrate or include other solvents used for crystallization. In some embodiments, the RNAi agent is a sodium salt. In some embodiments, the dsRNAi agent is in a salt form (e.g., a pharmaceutically acceptable salt form), where the salt is sodium (Na+), ammonium (NH4+), calcium (Ca2+), iron (Fe2+or Fe3+), magnesium (Mg2+), potassium (K+), pyridinium (C5H5NH+), quaternary ammonium (NR4+, R being an alkyl group or an aryl group as described herein), or copper (Cu2+).

[0238] In an aspect, the disclosure provides a dsRNA having sequences (e.g., antisense strand sequence) that can recognize a specific region of a HMGCR mRNA (e.g., human HMGCR mRNA) and lead cleavage of the HMGCR mRNA and silencing of the gene. The dsRNA includes a sense strand and an antisense strand and each strand may range from 12 to 30 nucleotides in length. In some embodiments, each strand may have 15 to 30 nucleotides in length. In some embodiments, each strand may have 15 to 25 nucleotides in length. In some embodiments, the antisense strand may have 15 to 25 nucleotides in length. In some embodiments, the sense strand may have 15 to 25 nucleotides in length. In some embodiments, the antisense strand may have 15 to 23 nucleotides in length. In some embodiments, the sense strand may have 15 to 23 nucleotides in length. In some embodiments, the antisense strand may have 18 to 25 nucleotides in length. In some embodiments, the sense strand may have 18 to 25 nucleotides in length.

[0239] In some embodiments, the sense strand may have 19 to 23 nucleotides in length. In some embodiments, the sense strand may have 21 to 23 nucleotides in length. In some embodiments, the sense strand may have 19 nucleotides in length. In some embodiments, the sense strand may have 20 nucleotides in length. In some embodiments, the sense strand may have 21 nucleotides in length. In some embodiments, the sense strand may have 22 nucleotides in length. In some embodiments, the sense strand may have 23 nucleotides in length. 53PAT059649-WO-PCT

[0240] In some embodiments, the antisense strand may have 19 to 25 nucleotides in length. In some embodiments, the antisense strand may have 19 to 23 nucleotides in length. In some embodiments, the antisense strand may have 21 to 23 nucleotides in length. In some embodiments, the antisense strand may have 23 to 25 nucleotides in length. In some embodiments, the antisense strand may have 19 nucleotides in length. In some embodiments, the antisense strand may have 20 nucleotides in length. In some embodiments, the antisense strand may have 21 nucleotides in length. In some embodiments, the antisense strand may have 22 nucleotides in length. In some embodiments, the antisense strand may have 23 nucleotides in length. In some embodiments, the antisense strand may have 24 nucleotides in length. In some embodiments, the antisense strand may have 25 nucleotides in length.

[0241] In some embodiments, the sense strand is 21 to 23 nucleotides in length and the antisense strand is 23 to 25 nucleotides in length. In some embodiments, the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length. In some embodiments, the sense strand is 22 nucleotides in length and the antisense strand is 24 nucleotides in length. In some embodiments, the sense strand is 23 nucleotides in length and the antisense strand is 25 nucleotides in length.

[0242] In an aspect, a dsRNA as described herein forms a double-stranded (or “duplex”) region made between a sense strand and an antisense strand and having 10 to 25 nucleotide pairs in length. The double stranded or duplex region are loaded into the RISC complex and subsequent specific degradation of the sense strand occurs during the RISC pathway. In some embodiments, the double stranded region has 10 nucleotide base pairs in length. In some embodiments, the double stranded region has 11 nucleotide base pairs in length. In some embodiments, the double stranded region has 12 nucleotide base pairs in length. In some embodiments, the double stranded region has 13 nucleotide base pairs in length. In some embodiments, the double stranded region has 14 nucleotide base pairs in length. In some embodiments, the double stranded region has 15 nucleotide base pairs in length. In some embodiments, the double stranded region has 16 nucleotide base pairs in length. In some embodiments, the double stranded region has 17 nucleotide base pairs in length. In some embodiments, the double stranded region has 18 nucleotide base pairs in length. In some embodiments, the double stranded region has 19 nucleotide base pairs in length. In some embodiments, the double stranded region has 20 nucleotide base pairs in length. In some embodiments, the double stranded region has 21 nucleotide base pairs in length. In 54PAT059649-WO-PCT some embodiments, the double stranded region has 22 nucleotide base pairs in length. In some embodiments, the double stranded region has 23 nucleotide base pairs in length.

[0243] In an aspect, a dsRNA as described herein may include at least one single- stranded nucleotide overhang, for example, for increasing in vivo effectiveness of the dsRNA and having substantially improved inhibition of the target genes. In certain aspects, the dsRNA may contain one or more extra nucleotides constituting overhang regions that locate other than the double stranded region at the 3′-end, 5′-end, or both ends of either stand or both strands (sense and antisense strands). In some embodiments, the overhang region may exist at the 3′-end, 5′-end, or both ends of the sense strand. In some embodiments, the overhang region may exist at the 3′-end, 5′-end, or both ends of the antisense strand. In some embodiments, the antisense strand may have a greater length than a length in the sense strand. In some embodiments, the antisense strand may have a shorter length than a length in the sense strand.

[0244] In some embodiments, the dsRNA may contain one or more extra nucleotides constituting overhang regions at the 3′-end, 5′-end, or both ends of the antisense strand. In some embodiments, the overhang region in the antisense strand may consist of 1-6 nucleotides in length, for example, 1 nucleotide, 2 nucleotides, 3 nucleotides, 4 nucleotides, 5 nucleotides, or 6 nucleotides in length. In some embodiments, the dsRNA may contain one or more extra nucleotides constituting overhang regions at the 3′-end, 5′-end, or both ends of the sense strand. In some embodiments, the overhang region may consist of 1-6 nucleotides in length, for example, 1 nucleotide, 2 nucleotides, 3 nucleotides, 4 nucleotides, 5 nucleotides, or 6 nucleotides in length.

[0245] In some embodiments, the antisense strand may include one-nucleotide overhang at the 5′ end. In some embodiments, the antisense strand may include one-nucleotide overhang at the 3′ end. In some embodiments, the antisense strand may include two- nucleotides overhang. In some embodiments, the antisense contains two-nucleotides overhang at the 5′ end. In some embodiments, the antisense contains two-nucleotides overhang at the 3′ end. In some embodiments, the antisense contains one-nucleotide overhang at the 5′ end and one-nucleotide overhang at the 3′ end. In some embodiments, the antisense strand may include three-nucleotide overhang. In some embodiments, the antisense contains three-nucleotides overhang at the 5′ end. In some embodiments, the antisense contains three-nucleotides overhang at the 3′ end. In some embodiments, the antisense contains two-nucleotides overhang at the 5′ end and one-nucleotide overhang at the 3′ end. In 55PAT059649-WO-PCT some embodiments, the antisense contains two nucleotides overhang at the 3′ end and one- nucleotide overhang at the 5′ end.

[0246] In certain aspects, a dsRNA as described herein may include at least one blunt end, e.g., for increasing in vivo stability with resistance to degradation in physiological surroundings. In some embodiments, the dsRNA may have a blunt end at the 3′-end, 5′-end, or both ends of the duplex. In some embodiments, the dsRNA includes one overhang (e.g., at 3′ end of antisense strand) and one blunt end (e.g., at 5′ end of antisense strand). In some embodiments, the dsRNA includes a blunt end at the 5′-end of the sense strand (and at 3′ end of the antisense strand) and contain overhang nucleotide(s) at the other end. In some embodiments, the dsRNA may have a blunt end at the 3′-end of the sense strand (and at 5′ end of the antisense strand) and contain overhang nucleotide(s) at the other end.

[0247] The sequences of the single strands (i.e., sense strand and antisense strand) of the dsRNA can be selected by selecting a target region and a length in the HMGCR mRNA. In certain aspects, a dsRNA as described herein may target a nucleotide region selected from regions of (i) 50-250 and (ii) 2400-2600 of a human HMGCR mRNA sequence that has at least about 85% (e.g., about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or 100%) identity to SEQ ID NO: 811 (human HMGCR isoform, transcript variant 1, mRNA (GenBank: NM_000859.3)). In some embodiments, the target region is selected from regions of (i) 100-200 and (ii) 2500-2600 of a human HMGCR mRNA sequence that has at least about 85% (e.g., about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or 100%) identity to SEQ ID NO: 811 (human HMGCR isoform, transcript variant 1, mRNA (GenBank: NM_000859.3)).

[0248] In certain aspects, an antisense strand of the dsRNA as described herein targets a nucleotide region selected from regions of (i) 50-250 and (ii) 2400-2600 of a human HMGCR mRNA sequence that has at least about 85% (e.g., about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or 100%) identity to SEQ ID NO: 811 (human HMGCR isoform, transcript variant 1, mRNA (GenBank: NM_000859.3)). In some embodiments, the antisense strand of the dsRNA as described herein targets a nucleotide region selected from regions of (i) 100-200 and (ii) 2500-2600 of a human HMGCR mRNA sequence that has at least about 85% (e.g., about 85%, about 86%, about 87%, about 88%, 56PAT059649-WO-PCT about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or 100%) identity to SEQ ID NO: 811 (human HMGCR isoform, transcript variant 1, mRNA (GenBank: NM_000859.3)).

[0249] In some embodiments, the antisense strand targets a region of 50-250th nucleotides in a human HMGCR mRNA sequence that has at least about 85% (e.g., about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or 100%) identity to SEQ ID NO: 811 (human HMGCR isoform, transcript variant 1, mRNA (GenBank: NM_000859.3)). In some embodiments, the antisense strand targets a region of 100-200th nucleotides in a human HMGCR mRNA sequence that has at least about 85% (e.g., about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or 100%) identity to SEQ ID NO: 811 (human HMGCR isoform, transcript variant 1, mRNA (GenBank: NM_000859.3)). In some embodiments, the antisense strand targets a region of 100-150th nucleotides in a human HMGCR mRNA sequence that has at least about 85% (e.g., about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or 100%) identity to SEQ ID NO: 811 (human HMGCR isoform, transcript variant 1, mRNA (GenBank: NM_000859.3)).

[0250] In some embodiments, the antisense strand targets a region of 2400-2600th nucleotides in a human HMGCR mRNA sequence that has at least about 85% (e.g., about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or 100%) identity to SEQ ID NO: 811 (human HMGCR isoform, transcript variant 1, mRNA (GenBank: NM_000859.3)). In some embodiments, the antisense strand targets a region of 2500-2600th nucleotides in a human HMGCR mRNA sequence that has at least about 85% (e.g., about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or 100%) identity to SEQ ID NO: 811 (human HMGCR isoform, transcript variant 1, mRNA (GenBank: NM_000859.3)). In some embodiments, the antisense strand targets a region of 2550-2600th nucleotides in a human HMGCR mRNA sequence that has at least about 85% (e.g., about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 57PAT059649-WO-PCT 98%, about 99% or 100%) identity to SEQ ID NO: 811 (human HMGCR isoform, transcript variant 1, mRNA (GenBank: NM_000859.3)).

[0251] In certain aspects, the target HMGCR mRNA sequence may range from 12 to 30 nucleotides, from 15 to 30 nucleotides, from 18 to 30 nucleotides, from 18 to 25 nucleotides, from 18 to 23 nucleotides. In some embodiments, the target HMGCR mRNA sequence may have 15 nucleotides in length. In some embodiments, the target HMGCR mRNA sequence may have 16 nucleotides in length. In some embodiments, the target HMGCR mRNA sequence may have 17 nucleotides in length. In some embodiments, the target HMGCR mRNA sequence may have 18 nucleotides in length. In some embodiments, the target HMGCR mRNA sequence may have 19 nucleotides in length. In some embodiments, the target HMGCR mRNA sequence may have 20 nucleotides in length. In some embodiments, the target HMGCR mRNA sequence may have 21 nucleotides in length. In some embodiments, the target HMGCR mRNA sequence may have 22 nucleotides in length. In some embodiments, the target HMGCR mRNA sequence may have 23 nucleotides in length.

[0252] In certain aspects, example dsRNA sequences including sense strands and antisense strands targeting the above indicated HMGCR mRNA (SEQ ID NO: 811, or GenBank: NM_000859.3) are in Table 1. Table 1 SEQ SEQ siRN position Sense Strand ID Antisense Strand ID A N58PAT059649-WO-PCT SEQ SEQ siRN position Sense Strand ID Antisense Strand ID A No NO: NO:59PAT059649-WO-PCT SEQ SEQ siRN position Sense Strand ID Antisense Strand ID A No NO: NO:60PAT059649-WO-PCT SEQ SEQ siRN position Sense Strand ID Antisense Strand ID A No NO: NO:61PAT059649-WO-PCT SEQ SEQ siRN position Sense Strand ID Antisense Strand ID A No NO: NO:62PAT059649-WO-PCT SEQ SEQ siRN position Sense Strand ID Antisense Strand ID A No NO: NO:63PAT059649-WO-PCT SEQ SEQ siRN position Sense Strand ID Antisense Strand ID A No NO: NO:64PAT059649-WO-PCT SEQ SEQ siRN position Sense Strand ID Antisense Strand ID A No NO: NO:65PAT059649-WO-PCT SEQ SEQ siRN position Sense Strand ID Antisense Strand ID A No NO: NO:66PAT059649-WO-PCT SEQ SEQ siRN position Sense Strand ID Antisense Strand ID A No NO: NO:67PAT059649-WO-PCT SEQ SEQ siRN position Sense Strand ID Antisense Strand ID A No NO: NO: 1234567(SEQ ID NO: 811) 1 ccttccgctc cgcgactgcg ttaactggag ccaggctgag cgtcggcgcc ggggttcggt 61 ggcctctagt gagatctgga ggatccaagg attctgtagc tacaatgttg tcaagacttt 121 ttcgaatgca tggcctcttt gtggcctccc atccctggga agtcatagtg gggacagtga 181 cactgaccat ctgcatgatg tccatgaaca tgtttactgg taacaataag atctgtggtt 241 ggaattatga atgtccaaag tttgaagagg atgttttgag cagtgacatt ataattctga 301 caataacacg atgcatagcc atcctgtata tttacttcca gttccagaat ttacgtcaac 361 ttggatcaaa atatattttg ggtattgctg gccttttcac aattttctca agttttgtat 421 tcagtacagt tgtcattcac ttcttagaca aagaattgac aggcttgaat gaagctttgc 481 cctttttcct acttttgatt gacctttcca gagcaagcac attagcaaag tttgccctca 541 gttccaactc acaggatgaa gtaagggaaa atattgctcg tggaatggca attttaggtc 601 ctacgtttac cctcgatgct cttgttgaat gtcttgtgat tggagttggt accatgtcag 661 gggtacgtca gcttgaaatt atgtgctgct ttggctgcat gtcagttctt gccaactact 721 tcgtgttcat gactttcttc ccagcttgtg tgtccttggt attagagctt tctcgggaaa 781 gccgcgaggg tcgtccaatt tggcagctca gccattttgc ccgagtttta gaagaagaag 841 aaaataagcc gaatcctgta actcagaggg tcaagatgat tatgtctcta ggcttggttc 901 ttgttcatgc tcacagtcgc tggatagctg atccttctcc tcaaaacagt acagcagata 961 cttctaaggt ttcattagga ctggatgaaa atgtgtccaa gagaattgaa ccaagtgttt 1021 ccctctggca gttttatctc tctaaaatga tcagcatgga tattgaacaa gttattaccc 1081 taagtttagc tctccttctg gctgtcaagt acatcttctt tgaacaaaca gagacagaat 1141 ctacactctc attaaaaaac cctatcacat ctcctgtagt gacacaaaag aaagtcccag 1201 acaattgttg tagacgtgaa cctatgctgg tcagaaataa ccagaaatgt gattcagtag 68PAT059649-WO-PCT 1261 aggaagagac agggataaac cgagaaagaa aagttgaggt tataaaaccc ttagtggctg 1321 aaacagatac cccaaacaga gctacatttg tggttggtaa ctcctcctta ctcgatactt 1381 catcagtact ggtgacacag gaacctgaaa ttgaacttcc cagggaacct cggcctaatg 1441 aagaatgtct acagatactt gggaatgcag agaaaggtgc aaaattcctt agtgatgctg 1501 agatcatcca gttagtcaat gctaagcata tcccagccta caagttggaa actctgatgg 1561 aaactcatga gcgtggtgta tctattcgcc gacagttact ttccaagaag ctttcagaac 1621 cttcttctct ccagtaccta ccttacaggg attataatta ctccttggtg atgggagctt 1681 gttgtgagaa tgttattgga tatatgccca tccctgttgg agtggcagga cccctttgct 1741 tagatgaaaa agaatttcag gttccaatgg caacaacaga aggttgtctt gtggccagca 1801 ccaatagagg ctgcagagca ataggtcttg gtggaggtgc cagcagccga gtccttgcag 1861 atgggatgac tcgtggccca gttgtgcgtc ttccacgtgc ttgtgactct gcagaagtga 1921 aagcctggct cgaaacatct gaagggttcg cagtgataaa ggaggcattt gacagcacta 1981 gcagatttgc acgtctacag aaacttcata caagtatagc tggacgcaac ctttatatcc 2041 gtttccagtc caggtcaggg gatgccatgg ggatgaacat gatttcaaag ggtacagaga 2101 aagcactttc aaaacttcac gagtatttcc ctgaaatgca gattctagcc gttagtggta 2161 actattgtac tgacaagaaa cctgctgcta taaattggat agagggaaga ggaaaatctg 2221 ttgtttgtga agctgtcatt ccagccaagg ttgtcagaga agtattaaag actaccacag 2281 aggctatgat tgaggtcaac attaacaaga atttagtggg ctctgccatg gctgggagca 2341 taggaggcta caacgcccat gcagcaaaca ttgtcaccgc catctacatt gcctgtggac 2401 aggatgcagc acagaatgtt ggtagttcaa actgtattac tttaatggaa gcaagtggtc 2461 ccacaaatga agatttatat atcagctgca ccatgccatc tatagagata ggaacggtgg 2521 gtggtgggac caacctacta cctcagcaag cctgtttgca gatgctaggt gttcaaggag 2581 catgcaaaga taatcctggg gaaaatgccc ggcagcttgc ccgaattgtg tgtgggaccg 2641 taatggctgg ggaattgtca cttatggcag cattggcagc aggacatctt gtcaaaagtc 2701 acatgattca caacaggtcg aagatcaatt tacaagacct ccaaggagct tgcaccaaga 2761 agacagcctg aatagcccga cagttctgaa ctggaacatg ggcattgggt tctaaaggac 2821 taacataaaa tctgtgaatt aaaaaagctc aatgcattgt cttgtggagg atgaatagat 2881 gtgatcactg agacagccac ttggtttttg gctctttcag agaggtctca ggttctttcc 2941 atgcagactc ctcagatctg aacacagttt agtgctttac atgctgtgct ctttgaagag 3001 atttcaacaa gaatattgta tgttaaagca tcagagatgg taatctacag ctcacctctg 3061 aaggcaaata taagctggga aaaaagtttt gatgaaattc ttgaagttca tggtgatcag 3121 tgcaattgac cttctccctc actcctgcca gttgaaaatg gatttttaaa ttatactgta 3181 gctgatgaaa ctcctgattt tgtagttaat ttattaagtc tgggatgtag aacttcaaga 3241 agtaagagct aagttctaag ttcatgtttg taaattaata cttcatttgg tgctggtcta 3301 ttttgatttt ggggggtaat cagcattatt cttcagaagg ggacctgttt tcttcaaggg 3361 aagaaacact cttattccca aactacagaa taatgtgtta aacatgctaa atagttctat 3421 caggaaaaca aatcactgta tttatctccg caggctattt gttcagagag gccttttgtt 3481 taaatataaa tgtttaaata taaatgtttg tctggattgg ctataacatg tctttcagca 3541 ttaggctttt aagaaacaca gggttttgta ttctttacta aagatatcag agctcttaat 3601 gttgcttaga tgagggtgac tgtcaagtac aagcaagact gggaccttag aaatcattgt 3661 agaaacacag ttttgaaaga aaaataccat gtctctaagc caactttaat tgcttaaaag 69PAT059649-WO-PCT 3721 acatttttat ttagttgaaa aatctagttt tttttgtaaa ctgtatcaaa tctgtatatg 3781 ttgtaataaa acttatgcta gtttattgga agtgttcaag aaataaaaat caacttgtgt 3841 actgataaaa tactctagcc tgggccagag aagataatgt tctttaatgt tgtccaggaa 3901 accctggctt gcttgccgag cctaatgaaa gggaaagtca gctttcagag ccagtgaagg 3961 agccacgtga atggccctag aactgtgcct agttcctgtg gccaggaggt tggtgactga 4021 aacattcaca cagggctctt tgatggaccc acgaacgctc ttagctttct cagggggtca 4081 gcagagttat tgaatcttaa ttttttttaa tgtacaagtt ttgtataaat aataaagaac 4141 tccttatttt gtattacatc taatgcttca agtgttgctc ttggaaagct gatgatgtct 4201 cttgtagaag atggactctg aaaaacattc caggaaacca tggcagcatg gagagcctct 4261 tagtgattgt gtctgcattg ttattgtgga agatttacct tttctgttgt acgtaaagct 4321 taaattgctt ttgttgtgac tttttagcca gtgacttttt ctgagctttt catggaagtg 4381 gcagtgaaaa atatgttgag tgttcatttt agtgactgta attaatatct tgctggatta 4441 atgttttgta caattactaa attgtataca ttttgttata gaatactttt ttctagtttc 4501 agtaaataat gaaaaggaag ttaataccaa

[0254] In Table 1, each code (letter, e.g., A, G, C, and U) represents a single ribonucleotide in the dsRNA. In some embodiments, the sequence list may be inclusive of any possible, additional modifications in a nucleobase, a ribose sugar ring, and / or a phosphate group (i.e., internucleoside linkage). In some embodiments, the last nucleotide from the 5′ end (or the first nucleotide from 3′ end) in each strand (sense strand and antisense strand) may have not include a phosphate group as being hydrolyzed or processed, e.g., during the synthesis of the oligonucleotides, but may contain 3′-terminal -OH group. In some embodiments, a phosphate group in the last nucleotide from the 5′ end (or the first nucleotide from 3′ end) in the sense strand may be added as a functional group for conjugation with a ligand.

[0255] In some embodiments, the dsRNA includes a sense strand having 10 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 10 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 11 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 11 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 12 contiguous nucleotides differing by no more 70PAT059649-WO-PCT than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 12 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 13 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 13 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 14 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 14 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 16 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 16 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 17 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 17 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 18 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 18 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected 71PAT059649-WO-PCT from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 19 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 19 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 20 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 20 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 21 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 21 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447.

[0256] In some embodiments, the dsRNA includes a sense strand having 10 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 10 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 11 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 11 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 12 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 12 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 13 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 72PAT059649-WO-PCT 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 13 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 14 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 14 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 15 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 15 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 16 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 16 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 17 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 17 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 18 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 18 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 19 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 19 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA 73PAT059649-WO-PCT includes a sense strand having 20 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 20 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 21 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 21 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447.

[0257] In some embodiments, the dsRNA includes a sense strand having 10 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 10 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 11 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 11 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 12 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 12 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 13 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 13 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 14 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 14 contiguous 74PAT059649-WO-PCT nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 15 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 15 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 16 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 16 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 17 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 17 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 18 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 18 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 19 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 19 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 20 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 20 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes a sense strand having 21 contiguous nucleotides differing by no more than 1 75PAT059649-WO-PCT nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440. In some embodiments, the dsRNA includes an antisense strand having 21 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes an antisense strand having 22 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes an antisense strand having 23 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447.

[0258] In some embodiments, the dsRNA includes (i) a sense strand having 15 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 15 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes (i) a sense strand having 16 contiguous nucleotides differing by no more than one, two or three from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 16 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes (i) a sense strand having 17 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 17 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes (i) a sense strand having 18 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 18 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes (i) a sense strand having 19 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 76PAT059649-WO-PCT 1 to 405 and 1434 to 1440 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 19 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes (i) a sense strand having 20 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 20 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447. In some embodiments, the dsRNA includes (i) a sense strand having 21 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 21 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447.

[0259] In certain aspects, when a sense strand or an antisense strand of a dsRNA in above paragraphs is differing by a certain number of nucleotides (e.g., one, two or three nucleotides) from a specific sequence (e.g., SEQ ID NOs: 1 to 810 and 1434 to 1447), it is meant by that the sense strand or the antisense strand of the dsRNA includes one, two or three nucleotides having different nucleobases compared to the nucleobases of the nucleotides at the corresponding positions of the specific sequence (e.g., SEQ ID NOs: 1 to 810 and 1434 to 1447). Modification Pattern

[0260] In an aspect, the disclosure provides a set of modification patterns determined or arranged by modified nucleotides in dsRNAs described herein. Aside from or in addition to the nucleobase sequences, various arrangements of modified nucleotides and the modification patterns thereof can be introduced, for example, to increase stability in a biological or physiological surrounding, to facilitate or promote cleavage by the RNA-induced silencing complex, and / or to mitigate or reduce off-targeting risk (e.g., to HMGCR off-targeting risk).

[0261] In an aspect, the disclosure provides a dsRNA that is partially (e.g., greater than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, or 45% of the total nucleotides), substantially (e.g., greater than about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of the total nucleotides), or entirely made of modified nucleotides, which can provide improved resistance to chemical and / or nuclease digestion and increased in vivo stability 77PAT059649-WO-PCT thereby imposing a longer in vivo half-life. Further, increasing the in vivo half-life of the dsRNA results in enhanced bioavailability and enhanced effectiveness in inhibiting expression or activity of a target gene (e.g., human HMGCR). For example, the stability of dsRNA in blood or serum may be determined, e.g., by its susceptibility to degradation by the cellular enzymes, which may be dependent on the characteristics (e.g., sequences, modification, modification pattern, or other chemical moieties) of each strand (i.e., sense strand or antisense strand) of the dsRNA. Thus, in certain aspect, the efficiency of dsRNA as a therapeutic agent may be improved by increasing the in vivo stability (e.g., in blood or serum) of the dsRNA while maintaining the ability of the dsRNA to mediate RNA interference in vivo. Modified Nucleotides

[0262] The modified nucleotides as used herein contain one or more modifications, for example, the modified nucleotides contain at least one chemical modification or replacement in an internucleoside linkage (“linkage”), a nucleobase, and / or a sugar moiety of the nucleotide. Non-limiting examples include a 2′-modification on a ribose sugar ring (e.g., 2′- deoxy, 2′-O-alkyl, 2′-halo, 2′-O-alkoxyalkyl, 2′-O-amino alkyl, etc.), 3′-modification (e.g., substitution) in backbone phosphate group, or 4′-modification on a ribose sugar ring (e.g., 4′- thio RNA). Also, other non-limiting examples of modifications may include one or more modifications selected from a deoxy modification, a 2′-O-alkyl modification, a 2′-halo modification, a 2′-5′-linkage modification, a conformationally restricting modification, an abasic modification, a 2′-amino-modification, a 2′-O-allyl modification, 2′-C-alkyl modification, a 2′-O-alkoxyalkyl modification, a morpholino modification, a modification containing a phosphoramidate group, a modification containing a non-natural nucleobase, a modification in a tetrahydropyran, a modification in a threose (TNA), a modification containing a 1,5-anhydrohexitol, a modification containing a cyclohexyl, a modification containing a cyclohexenyl, a modification containing a phosphorothioate group, a modification containing a methylphosphonate group, a modification containing an alkylphosphate, a modification containing a phosphonate, a modification containing an alkylphosphonate, a modification to form a thermally destabilizing nucleotide, a modification containing glycol (GNA), and a 2-O-(N-methylacetamide) modification. For example, a modified nucleotide may include a single modification, or two or more modifications at the positions at which the chemical modification groups do not hinder or intervene each other. 78PAT059649-WO-PCT

[0263] In some embodiments, each of the modified nucleotides is independently selected from LNA, GNA, TNA, 2′-O-alkoxyalkyl modified nucleotide, 2′-O-alkyl modified nucleotide, 2′-O-allyl modified nucleotide, 2′-C-allyl modified nucleotide, 2′-halo modified nucleotide, and 2′-deoxy modified nucleotide (DNA). The term alkyl, alkoxyl, allyl, amino, and halo can be interpreted as described above. In some embodiments, the modified nucleotides include at least one LNAs. In some embodiments, the modified nucleotides include at least one GNAs. In some embodiments, the modified nucleotides include at least one TNAs. In some embodiments, the modified nucleotides include at least one 2′-O- alkoxyalkyl modified nucleotides. In some embodiments, the modified nucleotides include at least one 2′-O-alkyl modified nucleotides. In some embodiments, the modified nucleotides include at least one 2′-O-allyl modified nucleotides. In some embodiments, the modified nucleotides include at least one 2′-C-allyl modified nucleotides. In some embodiments, the modified nucleotides include at least one 2′-halo (e.g., -F) modified nucleotides. In some embodiments, the modified nucleotides include at least one 2′-deoxy modified nucleotides (DNA).

[0264] In some embodiments, each of the modified nucleotides contain independently selected from LNA modification, GNA modification, TNA modification, 2′-O-alkoxyalkyl modification, 2′-O-alkyl modification, 2′-O-allyl modification, 2′-C-allyl modification, 2′- halo modification, and 2′-deoxy modification (DNA). The term alkyl, alkoxyl, allyl, amino, and halo can be interpreted as described above. In some embodiments, the modified nucleotides include at least one LNAs. In some embodiments, the modified nucleotides include at least one GNAs. In some embodiments, the modified nucleotides include at least one TNAs. In some embodiments, the modified nucleotides include at least one 2′-O- alkoxyalkyl modifications. In some embodiments, the modified nucleotides include at least one 2′-O-alkyl modifications. In some embodiments, the modified nucleotides include at least one 2′-O-allyl modifications. In some embodiments, the modified nucleotides include at least one 2′-C-allyl modifications. In some embodiments, the modified nucleotides include at least one 2′-halo (e.g., -F) modifications. In some embodiments, the modified nucleotides include at least one 2′-deoxy modifications (DNA).

[0265] In some embodiments, the modified nucleotide may be a bicyclic (or bridged) nucleic acid (“BNA”) having a covalent linkage between the 2′ and 4′ carbons on a ribose sugar. In some embodiments, the modified nucleotide is a locked RNA (“LNA”) having covalent linkage of a bicyclic sugar modification is a 4′-CH2-O-2′ linkage (methylene oxy), 79PAT059649-WO-PCTalso known as “LNA having a structur r a pharmaceuticallyacceptable salt thereof.

[0266] In some embodiments, a ribose r ng may be rep aced w t a glycol motif linked tophosphate and the GNA nucleotide has a structure of , or a pharmaceuticallyacceptable salt thereof.

[0267] In some embodiments, a ribose pentofuranosyl ring may be replaced with a threofuranosyl ring linked to the phosphate and a threofuranosyl nucleotide (TNA) may include a moiety a pharmaceutically acceptable salt thereof. In someembodiments, the TNA may have a structur a pharmaceutically acceptable salt thereof. In some embodiments, te in the TNA may be modified, e.g., with phosphorothioate group and modified TNA may include a structure of a pharmaceutically acceptable salt thereof. The TNA may further includetuents at 1′, 3′ and / or 4′ positions and such modified TNA may be encompassed by the definition of TNA herein. 80PAT059649-WO-PCT

[0268] In some embodiments, a ribose ring may not include a base and an abasicnucleotide has a structur a pharmaceutically acceptable salt thereof.

[0269] In certain asp eotide may include a heterocyclic group (e.g., 5 to 6 membered heterocycloalkyl ring) in place of a ribose ring. In some embodiments, the ribose ring may be replaced with a morpholinyl ring, e.g., to form an morpholino oligonucleotide. In some embodiments, the ribose ring may be replaced with an arabinose ring.

[0270] In certain aspects, the modified nucleotides contain one or more modification groups at 2′ position on the ribose ring by replacing 2′-OH. In some embodiments, the modification group may be hydrogen (i.e. deoxy), halogen (e.g., -F), substituted or unsubstituted alkyl (e.g., C1-C12 alkyl), or substituted or unsubstituted heteroalkyl (e.g., -O- (C1-C12alkyl), -N-(C1-C12alkyl), -C(O)NH-(C1-C12alkyl), -NHC(O)-(C1-C12alkyl), or - C(O)-(C1-C12 alkyl)). In some embodiments, the modification group may be hydrogen, -F, - O-alkyl (e.g., C1-C4alkyl), or -O-alkoxyalkyl (e.g., -O-(C1-C4alkylene)-(C1-C4alkoxyl)). Any of the alkyl, heteroalkyl, alkylene in the disclosure are optionally substituted with one or more of hydroxyl (-OH), C1-C3alkyl (e.g., methyl, or ethyl), amine (e.g., monoamine or diamine), alkoxyl (e.g., -O-CH3 (OMe) or -O-CH2CH3 (OEt)), halogen (e.g., -F) or the like.

[0271] In certain aspects, the modified nucleotides may include one or more of 2′-deoxy modification, 2′-O-alkyl modification, 2′-O-subsituted alkyl modification, 2′-O-alkoxyalkyl modification, and 2′-O-aminoalkyl modification. In some embodiments, the modified nucleotides may include one or more of 2′-deoxy modification, 2′-O-alkyl modification, 2′-O- subsituted alkyl modification, 2′-O-alkoxyalkyl modification, and 2′-O-aminoalkyl modification. In some embodiments, the modified nucleotides include at least one GNAs. In some embodiments, the modified nucleotides include at least one 2′-O-alkoxyalkyl modifications. In some embodiments, the modified nucleotides include at least one 2′-O- alkyl modifications. In some embodiments, the modified nucleotides include at least one 2′- O-allyl modifications. In some embodiments, the modified nucleotides include at least one 2′-C-allyl modifications. In some embodiments, the modified nucleotides include at least one 2′-halo (e.g., -F) modifications. In some embodiments, the modified nucleotides include at 81PAT059649-WO-PCT least one 2′-deoxy modifications (DNA). In some embodiments, the modified nucleotides do not include 2′-deoxy modifications (DNA).

[0272] In certain aspects, the modified nucleotides may include one or more of 2′-deoxy nucleotide (DNA), 2′-O-methyl (2′-OMe) modification, 2′-flouro (2′-F) modification, 2′-O- methoxyethyl (2′-O-MOE or “2′-MOE”) modification, 2′-O-aminopropyl (2′-O-AP) modification, 2′-O-dimethylaminoethyl (2′-O-DMAOE) modification, 2′-O- dimethylaminopropyl (2′-O-DMAP) modification, 2′-O-dimethylaminoethyloxyethyl (2′-O- DMAEOE) modification, and 2′-O-N-methylacetamido (2′-O-NMA) modification. In some embodiments, the modified nucleotides may include at least one 2′-deoxy modification (DNA). In some embodiments, the modified nucleotides may include at least one 2′-O- methyl (2′-OMe) modification. In some embodiments, the modified nucleotides may include at least one 2′-flouro (2′-F) modification. In some embodiments, the modified nucleotides may include at least one 2′-O-methoxyethyl (2′-O-MOE or “2′-MOE”) modification. In some embodiments, the modified nucleotides may include at least one 2′-O-aminopropyl (2′-O-AP) modification. In some embodiments, the modified nucleotides may include at least one 2′-O- dimethylaminoethyl (2′-O-DMAOE) modification. In some embodiments, the modified nucleotides may include at least one 2′-O-dimethylaminopropyl (2′-O-DMAP) modification. In some embodiments, the modified nucleotides may include at least one 2′-O- dimethylaminoethyloxyethyl (2′-O-DMAEOE) modification. In some embodiments, the modified nucleotides may include at least one 2′-O-N-methylacetamido (2′-O-NMA) modification.

[0273] In some embodiments, each modified nucleotide containing a modification on a 2′ sugar ring may optionally contain a phosphorothioate group at 5′ or 3′ linkage. In some embodiments, each modified nucleotide containing a modification on a 2′ sugar ring may optionally contain a modification such as an abasic modification or methylated nucleobase modification at nucleobase.

[0274] In certain aspects, the dsRNA is partially (e.g., greater than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, or 45% of the total nucleotides), substantially (e.g., greater than about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of the total nucleotides), or entirely made of modified nucleotides containing the modification on 2′ sugar ring. In some embodiments, the dsRNA is partially (e.g., greater than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, or 45% of the total nucleotides) made of modified nucleotides containing the modification on 2′ sugar ring. In some embodiments, the dsRNA 82PAT059649-WO-PCT is substantially (e.g., greater than about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of the total nucleotides) made of modified nucleotides containing the modification on 2′ sugar ring. In some embodiments, the dsRNA includes greater than about 80% of modified nucleotides containing the modification on 2′ sugar ring based on the total nucleotides. In some embodiments, the dsRNA includes greater than about 85% of modified nucleotides containing the modification on 2′ sugar ring based on the total nucleotides. In some embodiments, the dsRNA includes greater than about 90% of modified nucleotides containing the modification on 2′ sugar ring based on the total nucleotides. In some embodiments, the dsRNA includes greater than about 95% of modified nucleotides containing the modification on 2′ sugar ring based on the total nucleotides. In some embodiments, the dsRNA is entirely made of modified nucleotides containing the modification on 2′ sugar ring.

[0275] In certain aspects, the modified nucleotide may include a modification in a phosphate group or, in other words, an internucleoside linkage modification (e.g., phosphorothioate, phosphorodithioate, methylphosphonate, methylene phosphonate, or vinylphosphonate (VP) linkage). In some embodiments, the linkage modification may include phosphorothioate (PS) having a structure of , which may be an Rp isomer or an Sp isomer. In some embodiments, the linkage modcation may include phosphorothioate (PS) having a structure o , which may be a stereopure Rp isomer. In some embodiments, the linkagetion may include phosphorothioate (PS) having a structure of, which may be a stereopure Sp isomer. 83PAT059649-WO-PCT

[0276] For example, the modified nucleotide including 3′-PS modification can be represented wherein R represents H, OH or a substituent (e.g., -F, - CH3, -OMe, embodiments, the 3′-PS group may be a stereopure Spisomer. In some embodiments, the 3′-PS group may be a stereopure Rp isomer.

[0277] In certain aspects, the dsRNAi agent may be entirely made of modified nucleotides having one or more internucleoside linkage modification and / or modifications in the sugar moieties of the nucleotides. Example dsRNA (siRNA) with modified nucleotides, including sense strands and antisense strands targeting the above indicated HMGCR mRNA, are shown in Table 2. Table 2 siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID RNA NO NO: 3 4 5 6 784PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 8 9 0 1 2 3 4 5 6 7 885PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 9 0 1 2 3 4 5 6 7 8 986PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 0 1 2 3 4 5 6 7 8 9 087PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 1 2 3 4 5 6 7 8 9 0 188PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 2 3 4 5 6 7 8 9 0 1 289PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 3 4 5 6 7 8 9 0 1 2 390PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 4 5 6 7 8 9 0 1 2 3 491PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 5 6 7 8 9 0 1 2 3 4 592PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 6 7 8 9 0 1 2 3 4 5 693PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 7 8 9 0 1 2 3 4 5 6 794PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 8 9 0 1 2 3 4 5 6 7 895PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 9 0 1 2 3 4 5 6 7 8 996PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 0 1 2 3 4 5 6 7 8 9 097PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 1 2 3 4 5 6 7 8 9 0 198PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 2 3 4 5 6 7 8 9 0 1 299PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 3 4 5 6 7 8 9 0 1 2 3100PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 4 5 6 7 8 9 0 1 2 3 4101PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 5 6 7 8 9 0 1 2 3 4 5102PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 6 7 8 9 0 1 2 3 4 5 6103PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 7 8 9 0 1 2 3 4 5 6 7104PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 8 9 0 1 2 3 4 5 6 7 8105PAT059649-WO-PCT siRNA position SEQ SEQ No. in Sense Strand ID Antisense strand ID mRNA NO: NO: 9 0 1 2 3[ ] a e e ow s ows co es n t e nuc eot e sequences n a e an t e following Tables in the disclosure. Table A p: phosphate group / phosphodiester linkage p001: phosphorothioate linker(PS) SS: sense strand AS: antisense strand B001: abasic nucleoside A000: unmodified adenosine (A) G000: unmodified guanosine (G) A002: deoxyriboadenosine (dA) G002: deoxyriboguanosine (dG) A004: adenosine / 2′ OMe G004: guanosine / 2′ OMe A005: adenosine / 2′ MOE G005: guanosine / 2′ MOE A007: adenosine / 2′ F G007: guanosine / 2′ F A1016: GNA with adenine G1016: GNA with guanine A042: TNA with adenine G042: TNA with guanine A1017: locked nucleotide with adenine G1017: locked nucleotide with guanine X033A1027: adenosine / 5′(E)-VP-2′-OMe X033G1027: guanosine / 5′(E)-VP-2′-OMe C000: unmodified cytidine (C) U000: unmodified uridine (U) C002: deoxyribocytidine (dC) T000: ribothymidine C004: cytidine / 2′ OMe T002: deoxyribothymidine (dT) C005 or C005*: 5-methyl-cytidine / 2′ MOE U004: uridine / 2′ OMe C007: cytidine / 2′ F T005: ribothymidine (5-methyl uridine) / 2′ MOE C1016: GNA with cytosine U007: uridine / 2′ F C042: TNA with cytosine U1016: GNA with uracil C1017: locked nucleotide with cytosine U042: TNA with uracil X033C1027: cytidine / 5′(E)-VP-2′-OMe U1017: locked nucleotide with uracil X033U1027: uridine / 5′(E)-VP-2′-OMe 106PAT059649-WO-PCT

[0279] The sequences and sequence lists including modified nucleosides (e.g., RNA, RNA modified at a 2′-OH sugar moiety, or RNA modified at a nucleobase) in the disclosure (e.g., Tables 5-8, 10, and 14) are indicated with codes defined in Table A unless otherwise indicated. Each code consists of a letter representing a type of the nucleobase, e.g., “A”, “G”, “C”,“U,” or “T” and a numeric code representing a type of modification on a sugar ring.

[0280] For example, if a nucleoside is coded as “T005”, it is meant by a RNA nucleoside including a 2′-MOE sugar moiety and a thymine (or methylated uracil) as “T” indicates a thymine (or methylated uracil) nucleobase and “005” indicates a 2′-MOE substituent at 2′-OH position on the sugar ring.

[0281] In particular example, if a nucleoside is coded as “C005*”, it is meant by a RNA nucleoside including a 5-methylated cytosine and a 2′-MOE sugar moiety as “C” indicates a type of specific nucleobase, i.e.5-methylated cytosine, that can exist in combination with a 2′-MOE sugar moiety and “005” indicates a 2′-MOE substituent at 2′-OH position on the sugar ring.

[0282] The nucleosides in each sequence of the dsRNA are connected via phosphodiester group (“p” in Table A) or modification thereof (e.g., phosphorothioate linkage “p001” in Table A). In some embodiments, an example nucleotide may include a nucleoside and 3′- phosphodiester group. Example nucleotides may be presented in in Table A-1. Table A-1 Code for 5′-end first nucleotide Other position nucleotide107PAT059649-WO-PCT Code for 5′-end first nucleotide Other position nucleotide C004p:108PAT059649-WO-PCT Code for 5′-end first nucleotide Other position nucleotide G004p001:109PAT059649-WO-PCT Code for 5′-end first nucleotide Other position nucleotide A005p001:110PAT059649-WO-PCT Code for 5′-end first nucleotide Other position nucleotide U007p:111PAT059649-WO-PCT Code for 5′-end first nucleotide Other position nucleotide C007p001:112PAT059649-WO-PCT Code for 5′-end first nucleotide Other position nucleotide U1016p:113PAT059649-WO-PCT Code for 5′-end first nucleotide Other position nucleotide C042p:114PAT059649-WO-PCT Code for 5′-end first nucleotide Other position nucleotide G042p001: O115PAT059649-WO-PCT

[0283] In some embodiments, the dsRNA includes a sense strand having 10 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 10 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 11 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 11 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 12 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 12 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 13 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 13 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 14 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 14 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 16 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 16 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 17 contiguous nucleotides 116PAT059649-WO-PCT differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 17 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 18 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 18 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 19 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 19 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 20 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 20 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 21 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 21 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293.

[0284] In some embodiments, the dsRNA includes a sense strand having 10 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 10 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 11 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 11 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 12 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence 117PAT059649-WO-PCT selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 12 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 13 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 13 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 14 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 14 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 15 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 15 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 16 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 16 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 17 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 17 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 18 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 18 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 19 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 19 118PAT059649-WO-PCT contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 20 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 20 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 21 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 21 contiguous nucleotides differing by no more than 2 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293.

[0285] In some embodiments, the dsRNA includes a sense strand having 10 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 10 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 11 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 11 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 12 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 12 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 13 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 13 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 14 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 14 contiguous nucleotides differing by no more 119PAT059649-WO-PCT than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 15 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 15 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 16 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 16 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 17 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 17 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 18 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 18 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 19 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 19 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 20 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 20 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes a sense strand having 21 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052. In some embodiments, the dsRNA includes an antisense strand having 21 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes an 120PAT059649-WO-PCT antisense strand having 22 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes an antisense strand having 23 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293.

[0286] In some embodiments, the dsRNA includes (i) a sense strand having 15 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 15 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes (i) a sense strand having 16 contiguous nucleotides differing by no more than one, two or three from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 16 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes (i) a sense strand having 17 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 17 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes (i) a sense strand having 18 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 18 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes (i) a sense strand having 19 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 19 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes (i) a sense strand having 20 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 121PAT059649-WO-PCT 812 to 1052 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 20 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293. In some embodiments, the dsRNA includes (i) a sense strand having 21 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 812 to 1052 and (ii) an antisense strand forming a duplex with the sense strand of (i) and having 21 contiguous nucleotides differing by no more than one, two or three nucleotides from the nucleotide sequence selected from SEQ ID NOs: 1053 to 1293.

[0287] In certain aspects, when a sense strand or an antisense strand of a dsRNA in above paragraphs is differing by a certain number of nucleotides (e.g., one, two or three nucleotides) from a specific sequence (e.g., SEQ ID NOs: 812 to 1293), it is meant by that the sense strand or the antisense strand includes one, two or three nucleotides, having different nucleobases and / or different modifications compared to the nucleobases and / or the modifications of the nucleotides at the corresponding positions of the specific sequence (e.g., SEQ ID NOs: 812 to 1293). In some embodiments, when a sense strand or an antisense strand is differing by a certain number of nucleotides (e.g., one, two or three nucleotides) from a specific sequence (e.g., SEQ ID NOs: 812 to 1293), the sense strand or the antisense strand includes one, two, or three nucleotides, having different nucleobases compared to the nucleobases of the nucleotides at the corresponding positions of the specific sequence (e.g., SEQ ID NOs: 812 to 1293). In some embodiments, when a sense strand or an antisense strand is differing by a certain number of nucleotides (e.g., one, two or three nucleotides) from a specific sequence (e.g., SEQ ID NOs: 812 to 1293), the sense strand or the antisense strand includes one, two, or three nucleotides, having different modifications compared to the modifications of the nucleotides at the corresponding positions of the specific sequence (e.g., SEQ ID NOs: 812 to 1293). In some embodiments, when a sense strand or an antisense strand is differing by a certain number of nucleotides (e.g., one, two or three nucleotides) from a specific sequence (e.g., SEQ ID NOs: 812 to 1293), the sense strand or the antisense strand includes one, two, or three nucleotides having different nucleobases and different modifications compared to the nucleobases and the modifications of the nucleotides at the corresponding positions of the specific sequence (e.g., SEQ ID NOs: 812 to 1293).

[0288] In certain aspects, when a sense strand or an antisense strand of a dsRNA in above paragraphs is differing by a certain number of nucleotides (e.g., one, two or three nucleotides) from a specific sequence (e.g., SEQ ID NOs: 812 to 1293), it is meant by that 122PAT059649-WO-PCT the sense strand or the antisense strand includes one, two or three nucleotides, having different nucleobases, different modifications, and / or different phosphate linkages (e.g., phosphorothioate (PS)), compared to the nucleobases, the modifications, and / or the phosphate linkages of the nucleotides at the corresponding positions of the specific sequence (e.g., SEQ ID NOs: 812 to 1293). In some embodiments, when a sense strand or an antisense strand is differing by a certain number of nucleotides (e.g., one, two or three nucleotides) from a specific sequence (e.g., SEQ ID NOs: 812 to 1293), the sense strand or the antisense strand includes one, two, or three nucleotides, having different phosphate linkages compared to the phosphate linkages of the nucleotides at the corresponding positions of the specific sequence (e.g., SEQ ID NOs: 812 to 1293). In some embodiments, when a sense strand or an antisense strand is differing by a certain number of nucleotides (e.g., one, two or three nucleotides) from a specific sequence (e.g., SEQ ID NOs: 812 to 1293), the sense strand or the antisense strand includes one, two, or three nucleotides, having different nucleobases and different phosphate linkages compared to the nucleobases and the phosphate linkages of the nucleotides at the corresponding positions of the specific sequence (e.g., SEQ ID NOs: 812 to 1293). In some embodiments, when a sense strand or an antisense strand is differing by a certain number of nucleotides (e.g., one, two or three nucleotides) from a specific sequence (e.g., SEQ ID NOs: 812 to 1293), the sense strand or the antisense strand includes one, two, or three nucleotides, having different modifications and different phosphate linkages compared to the modifications and the phosphate linkages of the nucleotides at the corresponding positions of the specific sequence (e.g., SEQ ID NOs: 812 to 1293). In some embodiments, when a sense strand or an antisense strand is differing by a certain number of nucleotides (e.g., one, two or three nucleotides) from a specific sequence (e.g., SEQ ID NOs: 812 to 1293), the sense strand or the antisense strand includes one, two, or three nucleotides having different nucleobases, different modifications, and different phosphate linkages compared to the nucleobases, the modifications, and the phosphate linkages of the nucleotides at the corresponding positions of the specific sequence (e.g., SEQ ID NOs: 812 to 1293).

[0289] In certain aspects, the first nucleotide from the 5′ end of each strand (e.g., sense strand and antisense strand) may include an additional phosphate group or a variant thereof (e.g., phosphorothioate, phosphorodithioate, methylphosphonate, methylene phosphonate, or vinylphosphonate (VP)) attached or linked to the 5′ terminal group of the first nucleotide. 123PAT059649-WO-PCT

[0290] In some embodiments, the first nucleotide from the 5′ end of each strand (e.g., sense strand and antisense strand) includes a 5′-vinylphosphonate (5′-VP) group that is a chemical moiety having the structure of , or salts thereof, wherein represents the point of attachment to the 5′ carbon of the pentafuranosyl sugar. In some embodiments, the first nucleotide from the 5′ end of each strand (e.g., sense strand and antisense strand) may include (E)-vinylphosphonate (VP) having a structure o wherein represents the point of attachment to the 4′ carbon of the pentafura r. In some embodiments,the first nucleotide from the 5′ end of each strand (e.g., sense strand and antisense strand) may include (Z)-vinylphosphonate having a structure of wherein represents the point of attachment to the 4′ carbon of the pentafurr.

[0291] In certain aspects, one or more of the modified nucleotides contain a 2′ modification (e.g., 2′-OMe, 2′-F, 2′-MOE, 2′-deoxy, etc.) and an internucleoside linkage modification (e.g., phosphorothioate or (E)-vinylphosphonate). In some embodiments, one or more of the modified nucleotides contain 2′-OMe modification and phosphorothioate group. In some embodiments, one or more of the modified nucleotides contain 2′-OMe modification and (E)-vinylphosphonate group. In some embodiments, one or more of the modified nucleotides contain 2′-F modification and phosphorothioate group. In some embodiments, one or more of the modified nucleotides contain 2′-F and (E)-vinylphosphonate group. In some embodiments, one or more of the modified nucleotides contain 2′-MOE modification and phosphorothioate group. In some embodiments, one or more of the modified nucleotides contain 2′-MOE modification and (E)-vinylphosphonate group. In some embodiments, one or more of the modified nucleotides contain 2′-deoxy modification and phosphorothioate group. In some embodiments, one or more of the modified nucleotides contain 2′-OMe modification and (E)-vinylphosphonate group. In some embodiments, one or more of the modified nucleotides are GNA containing (E)-vinylphosphonate group. In some embodiments, one or more of the modified nucleotides are GNA containing a phosphorothioate group.

[0292] In certain aspects, the modified nucleotides contain one or more modifications on a modified nucleobase. In some embodiments, one or more of the modified nucleotides may 124PAT059649-WO-PCT include thymine (“T”) nucleobase (“ribothymidine” or “5-methyluridine”) in the ribonucleotide (e.g., including 2′-OH). In some embodiments, one or more of the modified nucleotides may include methylcytosine nucleobase (e.g., 5-methylcytidine or N4- methylcytidine). In certain aspects, one or more of the modified nucleotides may contain no nucleobase or be abasic. Sense Strand (SS)

[0293] In certain aspects, a sense strand of the dsRNA agent(s) as described herein are substantially (e.g., greater than about 80%, 85%, 90%, or 95% of the total nucleotides) made of modified nucleotides. In another certain aspect, the sense strand is entirely made of modified nucleotides.

[0294] In certain aspects, a sense strand of the dsRNA agent(s) as described herein includes two or more 2′-MOE modifications. In some embodiments, the sense strand includes two, four, six or eight 2′-MOE modifications. In some embodiments, the sense strand includes two 2′-MOE modifications. In some embodiments, the sense strand includes four 2′-MOE modifications. In some embodiments, the sense strand includes six 2′-MOE modifications. In some embodiments, the sense strand includes eight 2′-MOE modifications.

[0295] In some embodiments, the 2′-MOE modified nucleotides in the sense strand as described herein include a structur r a pharmaceutically acceptable salt thereof, whereinis an attachment point to a linkage (e.g., phosphate or phosphorothioate group) or thejacent nucleotides and “Base” is a nucleobase.

[0296] In some embodiments, the 2′-MOE modified nucleotides in the sense strand as described herein include a structur r apharmaceutically acceptable salt thereof, wherein is an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacent nucleotides and “Base” is a nucleobase. In some 125PAT059649-WO-PCT embodiments, the 2′-MOE modified nucleotides in the sense strand as described herein include a structure a pharmaceutically acceptable salt thereof.

[0297] In som E modified nucleotides include a structure ofalt, , , , he adjacent nucleoes. In some embodiments, the 2′-MOE modified nucleotides include a llyclude a structure a pharmaceutically acceptable salt thereof.126PAT059649-WO-PCT

[0298] In some embodiments, the 2′-MOE modified nucleotides include a nucleotide r ato a terminal group (e.g., H, OH, or salt) or the adjacent nucleotides. In some embodiments, the 2′-MOE modified nucleotides include a nucleotide having a structu , or a pharmaceutically acceptable salt thereof. In some embodnucleotides include a nucleotide having a structur ,a pharmaceutically acceptable salt thereof. In some127PAT059649-WO-PCT embodiments, the 2′-MOE modified nucleotides include a nucleotide having a structure of a pharmaceutically acceptable salt thereof. ments, the 2′-MOE modified nucleotides in the sense strand asdescribed herein include a nucleotide having a structur ,a pharmaceutically acceptable salt thereof, wherein isroup (e.g., H, OH, or salt) or the adjacent nucleots. In some embodiments, the 2′-MOE modified nucleotides in the sense strand as described herein include a nucleotide having a structur , or a pharmaceutically acceptable salt there. 128PAT059649-WO-PCT

[0300] In some embodiments, the 2′-MOE modified nucleotides include a nucleotide having a structure r a pharmaceuticallyterminal group (e.g., H, OH, or salt) or the adjacent nucle des. In some embodiments, the 2′-MOEmodified nucleotides include a nucleotide having a structu , or a pharmaceutically acceptable salt thereof.

[0301] In certain aspects, at least one of the 2′-MOE modified nucleotides in the sense O strand as described herein has a structur ,a pharmaceutically acceptable salt thereof, wherein is anrminal group (e.g., H, OH, or salt) or the adjacentnucleotides. In 129PAT059649-WO-PCT some embodiments, at least one of the 2′-MOE modified nucleotides in the sense strand as described herein has a structur r a pharmaceutically acceptable salt thereof.

[0302] In some embodiments, the first nucleotide from the 5′ end of the sense strand includes a structure r a pharmaceutically acceptable salt ther tide from the 5′ end of thesense strand includes a structur r a pharmaceutically acceptable salt thereof.

[0303] In some embodiments, the first nucleotide from the 3′ end of the sense strand includes a structure r a pharmaceutically acceptable salt ther, m the 3′ end of the 130PAT059649-WO-PCT or, . In some embodiments, the first nucleotide from thend of the sense strand includes a a pharmaceutically acceptable salt thereof.one of the 2′-MOE modified nucleotides in the sense strand as described herein has a structur ,131PAT059649-WO-PCT NH2N a pharmaceutically acceptable salt thereof, wherein is an nal group (e.g., H, OH, or salt) or the adjacent nu otides. Insome embodiments, at least one of the 2′-MOE modified nucleotides in the sense strand as described herein has a structur r a pharmaceutically acceptable salt thereof.

[0305] In some embodiments, the first nucleotide from the 5′ end of the sense strand llye sense strand includes a structur r a pharmaceutically acceptable salt thereof.132PAT059649-WO-PCT

[0306] In some embodiments, the first nucleotide from the 3′ end of the sense strand llysense strand includes a structur ,,a pharmaceutically acceptable salt thereof, wherein is and. In some embodiments, the first nucleotide from the 3′ end of the sense strand includes a structur r a pharmaceutically acceptable salt thereof.133PAT059649-WO-PCT

[0307] In certain aspects, at least one of the 2′-MOE modified nucleotides in the sense strand as described herein has a structurr a pharmaceutically acceptable, al group (e.g., H, OH, or salt) or the adjacent nucleoties. In some embodiments, at least one of the 2′-MOE modified nucleotides in the sense strand as described herein has a structure of a pharmaceutically acceptable salt thereof.ents, the first nucleotide from the 5′ end of the sense strand r acleotide from the 134PAT059649-WO-PCT r a

[0309] In some embodiments, the first nucleotide from the 3 end of the sense strand has ally, ,135PAT059649-WO-PCT a pharmaceutically acceptable salt thereof, wherein is an n some embodiments, the first nucleotide from th ′ end ofthe sense strand has a structur lly acceptable salt thereof.

[0310] In certain aspects, at least one of the 2′-MOE modified nucleotides in the sense llyacceptable salt thereof, wherein is an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacent nucleotides. In some embodiments, at least one of the 2′-MOE modified 136PAT059649-WO-PCT nucleotides in the sense strand as described herein has a structure of a pharmaceutically acceptable salt thereof. the first nucleotide from the 5′ end of the sense strandr a from the5′ end of the sense strand includes a structur , or a pharmaceutically acceptable salt thereof.

[0312] In some embodiments, the first nucleotide from the 3′ end of the sense strand has r ae from the 3′ end of the sense strand has a structure of 137PAT059649-WO-PCT r a pharmaceuticallyaccep a e sa e eo , w e e s a a ac e po o a ga d. In some embodiments, the first nucleotide from the 3′d of the sense strand has a structure of a pharmaceutically acceptable salt thereof.2′-MOE modified nucleotides locate at both 5′ and 3′ ends of a sense strand so as to form a structural confinement (“2′-MOE clamp”) at the sense strand termini. In some embodiments, the 2′-MOE clamps may be symmetric and having the same number of 2′-MOE modified nucleotides at both 5′ and 3′ ends of the sense strand. For example, the sense strand includes one 2′-MOE modified nucleotide at 5′ end and one 2′- MOE modified nucleotide at 3′ end; two 2′-MOE modified nucleotides at 5′ end and two 2′- MOE modified nucleotides at 3′ end; or three 2′-MOE modified nucleotides at 5′ end and three 2′-MOE modified nucleotides at 3′ end. In some embodiments, the 2′-MOE clamps 138PAT059649-WO-PCT may be asymmetric and having different numbers of 2′-MOE nucleotides at 5′ and 3′ ends of the sense strand. For example, the sense strand includes one 2′-MOE modified nucleotide at 5′ end only; one 2′-MOE modified nucleotide at 3′ end only; two 2′-MOE modified nucleotides at 5′ end only; two 2′-MOE modified nucleotides at 3′ end only; one 2′-MOE modified nucleotide at 5′ end and two 2′-MOE modified nucleotides at 3′ end; or two 2′-MOE modified nucleotides at 5′ end and one 2′-MOE modified nucleotide at 3′ end.

[0314] In certain aspects, the sense strand includes one 2′-MOE modified nucleotide at 5′ end and one 2′-MOE modified nucleotide at 3′ end. In some embodiments, the sense strand includes only one 2′-MOE modified nucleotide at 5′ end and only one 2′-MOE modified nucleotide at 3′ end. In some embodiments, the sense strand includes only one 2′-MOE modified nucleotide at 5′ end. In some embodiments, the sense strand includes only one 2′- MOE modified nucleotide at 3′ end.

[0315] In certain aspects, the sense strand includes at least two contiguous 2′-MOE modified nucleotides at 5′ end and at least two 2′-MOE modified nucleotides at 3′ end. In some embodiments, the sense strand includes only two 2′-MOE modified nucleotides at 5′ end and only two 2′-MOE modified nucleotides at 3′ end. In some embodiments, the sense strand includes only two 2′-MOE modified nucleotides at 5′ end. In some embodiments, the sense strand includes only two 2′-MOE modified nucleotides at 3′ end.

[0316] In certain aspects, the sense strand is 21 nucleotides in length. In some embodiments, the sense strand includes one, two, three, or four 2′-MOE modified nucleotides positioned at the 1st, 2nd, 20th, and / or 21st nucleotides from the 5′ end of the sense strand. In some embodiments, the sense strand includes two 2′-MOE modified nucleotides positioned at the 1st, 2nd, 20th, or 21st nucleotides from the 5′ end of the sense strand. In some embodiments, the sense strand includes three 2′-MOE modified nucleotides positioned at the 1st, 2nd, 20th, or 21st nucleotides from the 5′ end of the sense strand. In some embodiments, the sense strand includes 2′-MOE modified nucleotides positioned at the 1st, 2nd, 20th, and 21st nucleotides from the 5′ end of the sense strand. In some embodiments, the sense strand does not include a 2′-MOE modified nucleotide at the 3rd to 19th nucleotides from 5′ end of the sense strands.

[0317] Alternatively, in certain aspects, a sense strand of the dsRNA as described herein includes two or more TNAs. In some embodiments, the sense strand includes two, four, six or eight TNAs. In some embodiments, the sense strand includes two TNAs. In some 139PAT059649-WO-PCT embodiments, the sense strand includes four TNAs. In some embodiments, the sense strand includes six TNAs. In some embodiments, the sense strand includes eight TNAs.

[0318] In certain aspects, the sense strand includes at least two contiguous TNAs at 5′ end and at least two TNAs at 3′ end. In some embodiments, the sense strand includes only two TNAs at 5′ end and only two TNAs at 3′ end. In some embodiments, the sense strand includes only two TNAs at 5′ end. In some embodiments, the sense strand includes only two TNAs at 3′ end.

[0319] In some embodiments, the TNAs in the sense strand as described herein include a a pharmaceutically acceptable salt thereof, wherein is .g., phosphate or phosphorothioate group) or the acentnucleotides and “Base” is a nucleobase.

[0320] In some embodiments, the TNAs in the sense strand as described herein include a structure a pharmaceutically acceptable salt thereof, were n s an attac ment pont to a terminal group (e.g., H, OH, or salt) or the adjacent nucleoes and “Base” is a nucleobase. In some embodiments, the TNAs in the dsRNA as described herein include a structur r a pharmaceutically acceptable salt thereof.140PAT059649-WO-PCT

[0321] In some embodiments, the TNAs include a structu ,a pharmaceutically acceptable salt thereof,al group (e.g., H, OH, or salt) or the adjacent nucleotides. In some embodiments, the TNAs include a structu , or a pharmaceutically acceptable salt thereof.

[0322] In some embodiments, the TNAs include a nucleotide having a structure of llyacceptable salt thereof, wherein is an attachment point to a terminal group (e.g., H, OH, or141PAT059649-WO-PCT salt) or the adjacent nucleotides. In some embodiments, the TNAs include a nucleotide a pharmaceutically acceptable salt thereof. n the dsRNA as described herein include anucleotide having a structur ,a pharmaceutically acceptable salt thereof, wherein is anroup (e.g., H, OH, or salt) or the adjacent nucleots. In some embodiments, the TNAs in the dsRNA as described herein include a nucleotide having a structure a pharmaceutically acceptable salt thereof.

[0324] include a nucleotide having a structure of r a142PAT059649-WO-PCT pharmaceutically acceptable salt thereof, wherein is an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacent nucle ides. In some embodiments, the TNAs include a nucleotide having a structur r a pharmaceutically acceptable salt thereof.

[0325] In certain aspects, at leastone of the TNAs in the sense strand as described herein rap y p , p minal group (e.g., H, OH, or salt) or the adjacent nucledes. In some embodiments, at least one of the TNAs in the sense strand as described herein has a structu r a pharmaceutically acceptable salt thereof.

[0326] In some embodiments, the first nucleotide from the 5′ end of the sense strand includes a structure r a pharmaceutically acceptable salt ther, e from the 5′ end of the 143PAT059649-WO-PCT sense strand includes a structur a pharmaceutically acceptable salt thereof.

[0327] In some embodiments, the first nucleotide from the 3′ end of the sense strand includes a structure lly acceptable salt ther of ther apharmaceutically acceptable salt thereof and is an attachment point to a ligand. In some144PAT059649-WO-PCT embodiments, the first nucleotide from the 3′ end of the sense strand includes a structure of , or a pharmaceutically acceptable salt thereof. in aspects, at least one of the TNAs in the sense strand as described hereinr a, group (e.g., H, OH, or salt) or the adjacent nucledes. In some embodiments, at least one of the TNAs in the sense strand as described herein has a structu , or a pharmaceutically acceptable salt thereof.

[0329] In some embodiments, the first nucleotide from the 5′ end of the sense strand includes a structure r a pharmaceutically acceptable salt therfrom the 5′ end of the 145PAT059649-WO-PCT sense strand includes a structur r a pharmaceutically acceptable salt thereof.

[0330] In some embodiments, the first nucleotide from the 3′ end of the sense strand includes a structure lly acceptable salt ther,or a pharmaceutically acceptable salt thereof and is an attachment point to a ligand. In some embodiments, the first nucleotide from th′ end of the sense strand includes a a pharmaceutically acceptable salt thereof.146PAT059649-WO-PCT

[0331] In certain aspects, at least one of the TNAs in the sense strand as described herein ,orgroup (e.g., H, OH, or salt) or the adjacent nucleotides. In some embodiments, at least one of the TNAs in the sense strand as described herein has a structu , or a pharmaceutically acceptable salt thereof.

[0332] In some embodiments, the first nucleotide from the 5′ end of the sense strand includes a structure r a pharmaceutically acceptable salt therm the 5′ end of the sense strand includes a structur r a pharmaceutically acceptable salt thereof.147PAT059649-WO-PCT

[0333] In some embodiments, the first nucleotide from the 3′ end of the sense strand r a ucleotide from thealtthereof and is an attachment point to a ligand. In some embodiments, the first nucleotidefrom the 3′ end of the sense strand includes a structur r a pharmaceutically acceptable salt thereof.148PAT059649-WO-PCT

[0334] In certain aspects, at least one of the TNAs in the sense strand as described herein ,a pharmaceutically acceptable salt thereof, wherein is an roup (e.g., H, OH, or salt) or the adjacent nucleots. Insome embodiments, at least one of the TNAs in the sense strand as described herein has a a pharmaceutically acceptable salt thereof.nucleotide from the 5′ end of the sense strand includes a structure r a pharmaceutically aotide from the 149PAT059649-WO-PCT 5′ end of the sense strand includes a structur , or a pharmaceutically acceptable salt thereof.

[0336] In some embodiments, the first nucleotide from the 3 end of the sense strand r a from theblesalt thereof and is an attachment point to a ligand. In some embodiments, the first 150PAT059649-WO-PCT nucleotide from the 3′ end of the sense strand includes a structure of , or a pharmaceutically acceptable salt thereof. cts, the TNAs locate at both 5′ and 3′ ends of a sense strand so as toform a structural confinement (“TNA clamp”) at the sense strand termini. In some embodiments, the TNA clamps may be symmetric and having the same number of TNAs at both 5′ and 3′ ends of the sense strand. For example, the sense strand includes one TNA at 5′ end and one TNA at 3′ end; two TNAs at 5′ end and two TNAs at 3′ end; or three TNAs at 5′ end and three TNAs at 3′ end. In some embodiments, the TNA clamps may be asymmetric and having different numbers of TNAs at 5′ and 3′ ends of the sense strand. For example, the sense strand includes one TNA at 5′ end only; one TNA at 3′ end only; two TNAs at 5′ end only; two TNAs at 3′ end only; one TNA at 5′ end and two TNAs at 3′ end; or two TNAs at 5′ end and one TNA at 3′ end.

[0338] In certain aspects, the sense strand includes one TNA at 5′ end and one TNA at 3′ end. In some embodiments, the sense strand includes only one TNA at 5′ end and only one TNA at 3′ end. In some embodiments, the sense strand includes only one TNA at 5′ end. In some embodiments, the sense strand includes only one TNA at 3′ end.

[0339] In certain aspects, the sense strand includes at least two contiguous TNAs at 5′ end and at least two TNAs at 3′ end. In some embodiments, the sense strand includes only two TNAs at 5′ end and only two TNAs at 3′ end. In some embodiments, the sense strand includes only two TNAs at 5′ end. In some embodiments, the sense strand includes only two TNAs at 3′ end.

[0340] In certain aspects, the sense strand is 21 nucleotides in length. In some embodiments, the sense strand includes one, two, three, or four TNAs positioned at the 1st, 2nd, 20th, and / or 21st nucleotides from the 5′ end of the sense strand. In some embodiments, the sense strand includes two TNAs positioned at the 1st, 2nd, 20th, or 21st nucleotides from the 5′ end of the sense strand. In some embodiments, the sense strand includes three TNAs positioned at the 1st, 2nd, 20th, or 21st nucleotides from the 5′ end of the sense strand. In some embodiments, the sense strand includes TNAs positioned at the 1st, 2nd, 20th, and 21st nucleotides from the 5′ end of the sense strand. 151PAT059649-WO-PCT

[0341] In certain aspects, the sense strand of the dsRNA as described herein includes two or more 2′-F modifications. In some embodiments, the sense strand of the dsRNA includes two, three, four, five, six, seven, or eight 2′-F modified nucleotides. In some embodiments, the sense strand includes two 2′-F modified nucleotides. In some embodiments, the sense strand includes three 2′-F modified nucleotides. In some embodiments, the sense strand includes four 2′-F modified nucleotides. In some embodiments, the sense strand includes five 2′-F modified nucleotides. In some embodiments, the sense strand includes six 2′-F modified nucleotides. In some embodiments, the sense strand includes seven 2′-F modified nucleotides. In some embodiments, the sense strand includes eight 2′-F modified nucleotides. In some embodiments, two contiguous 2′-F modified nucleotides locate in the sense strand. In some embodiments, three contiguous 2′-F modified nucleotides locate in the sense strand. In some embodiments, four contiguous 2′-F modified nucleotides locate in the sense strand.

[0342] In certain aspects, the sense strand is 21 nucleotides in length. In some embodiments, 2′-F modified nucleotides locate at 5th, 7th, 8th, and / or 9th positions from the 5′ end of the sense strand. In some embodiments, 2′-F modified nucleotides locate at 6th, 8th, 9th, and / or 10th positions from the 5′ end of the sense strand. In some embodiments, 2′-F modified nucleotides locate at 7th, 9th, 10th, and / or 11th positions from the 5′ end of the sense strand. In some embodiments, 2′-F modified nucleotides locate at 8th, 10th, 11th, and / or 12th positions from the 5′ end of the sense strand. In some embodiments, 2′-F modified nucleotides locate at 9th, 11th, 12th, and / or 13th positions from the 5′ end of the sense strand.

[0343] In some embodiments, the sense strand includes 2′-OMe modified nucleotides in the remaining positions in the sense strand.

[0344] In certain aspects, the sense strand includes one to six (e.g., 1, 2, 3, 4, 5 or 6) phosphorothioate (PS) linkages between nucleosides. In some embodiments, the sense strand includes one, two, three, or four phosphorothioate (PS) linkages between nucleosides.

[0345] In certain aspects, the sense strand is 21 nucleotides in length. In some embodiments, the sense strand includes two 3′-PS modified nucleotides at the 1st, 2nd, 19th and / or 20th positions from 5′-end of the sense strand. In some embodiments, the sense strand includes three 3′-PS modified nucleotides at the 1st, 2nd, 19th and / or 20th positions from 5′- end of the sense strand. In some embodiments, the sense strand includes 3′-PS modified nucleotides at the 1st, 2nd, 19th and 20th positions from 5′-end of the sense strand. 152PAT059649-WO-PCT

[0346] In certain aspects, the sense strand is 21 nucleotides in length. In some embodiments, the sense strand includes 3′-PS modified nucleotides at the 1st and 2nd positions from 5′-end of the sense strand. In some embodiments, the sense strand includes a 3′-PS modified nucleotide at the 1st position from 5′-end of the sense strand. In some embodiments, the sense strand includes 3′-PS modified nucleotides at the 1st and 20th positions from 5′-end of the sense strand.

[0347] In certain aspects, the sense strand includes two to eight phosphorothioate (PS) groups or linkages between nucleosides. In certain aspects, the sense strand is 21 nucleotides in length. In some embodiments, the sense strand includes two 3′-PS modified nucleotides positioned at the 1st, 2nd, 3rd, 4th, 17th, 18th, 19th and / or 20th nucleotides from 5′-end of the sense strand. In some embodiments, the sense strand includes four 3′-PS modified nucleotides positioned at the 1st, 2nd, 3rd, 4th, 17th, 18th, 19th and / or 20th nucleotides from 5′-end of the sense strand. In some embodiments, the sense strand includes six 3′-PS modified nucleotides positioned at the 1st, 2nd, 3rd, 4th, 17th, 18th, 19th and / or 20th nucleotides from 5′-end of the sense strand. In some embodiments, the sense strand includes 3′-PS modified nucleotides positioned at the 1st, 2nd, 3rd, 4th, 17th, 18th, 19th and 20th nucleotides from 5′-end of the sense strand.

[0348] In certain aspects, the sense strand is 21 nucleotides in length. In some embodiments, at least one of the 3′-PS groups at the 1st, 2nd, 3rd, 4th, 17th, 18th, 19th and / or 20th nucleotides from 5′-end of the sense strand is a stereopure Rp isomer. In some embodiments, at least one of the 3′-PS groups at the 1st, 2nd, 19th and / or 20th nucleotides from 5′-end of the sense strand is a stereopure Rp isomer. In some embodiments, at least one of the 3′-PS groups at the 1st and / or 20th nucleotides from 5′-end of the sense strand is a stereopure Rp isomer. In some embodiments, the 3′-PS group at the 1st nucleotide from 5′- end of the sense strand is a stereopure Rp isomer. In some embodiments, the 3′-PS group at the 2nd nucleotide from 5′-end of the sense strand is a stereopure Rp isomer. In some embodiments, the 3′-PS group at the 19th nucleotide from 5′-end of the sense strand is a stereopure Rp isomer. In some embodiments, the 3′-PS group at the 20th nucleotide from 5′- end of the sense strand is a stereopure Rp isomer. In some embodiments, the 3′-PS groups at the 1st and 20th nucleotides from 5′-end of the sense strand are stereopure Rp isomers. In some embodiments, the 3′-PS groups at the 1st, 2nd, 19thand 20th nucleotides from 5′-end of the sense strand are stereopure Rp isomers. 153PAT059649-WO-PCT

[0349] In certain aspects, the sense strand is 21 nucleotides in length. In some embodiments, at least one of the 3′-PS groups at the 1st, 2nd, 3rd, 4th, 17th, 18th, 19th and / or 20th nucleotides from 5′-end of the sense strand is a stereopure Sp isomer. In some embodiments, at least one of the 3′-PS groups at the 1st, 2nd, 19th and / or 20th nucleotides from 5′-end of the sense strand is a stereopure Sp isomer. In some embodiments, at least one of the 3′-PS groups at the 1st and / or 20th nucleotides from 5′-end of the sense strand is a stereopure Sp isomer. In some embodiments, the 3′-PS group at the 1st nucleotide from 5′- end of the sense strand is a stereopure Sp isomer. In some embodiments, the 3′-PS group at the 2nd nucleotide from 5′-end of the sense strand is a stereopure Sp isomer. In some embodiments, the 3′-PS group at the 19th nucleotide from 5′-end of the sense strand is a stereopure Sp isomer. In some embodiments, the 3′-PS group at the 20nd nucleotide from 5′- end of the sense strand is a stereopure Sp isomer. In some embodiments, the 3′-PS groups at the 1st and 20th nucleotides from 5′-end of the sense strand are stereopure Sp isomers. In some embodiments, the 3′-PS groups at the 1st, 2nd, 19th, and 20th nucleotides from 5′-end of the sense strand are stereopure Sp isomers.

[0350] In certain aspects, a sense strand of the dsRNA as described herein includes one or more of 2′-MOE modified nucleotides, one or more of 2′-F modified nucleotides, and one or more of 2′-OMe modified nucleotides. In certain aspects, a sense strand of the dsRNA as described herein consists of 2′-MOE modified nucleotides, 2′-F modified nucleotides and 2′- OMe modified nucleotides.

[0351] In certain aspects, a sense strand of the dsRNA as described herein may have a Formula (I), 5′-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-3′ (I) wherein: each X1 to X21 is independently a nucleotide, each X1, X2,X20, and X21is a 2′-MOE modified nucleotide; each X3 to X19 is independently selected from a deoxyribonucleotide, 2′-MOE modified nucleotide, 2′-F modified nucleotide, and 2′-OMe modified nucleotide.

[0352] In some embodiments, the first nucleotide from the 5′ end of the sense strand (X1) is a 2′-MOE modified nucleotide with a nucleobase T. In some embodiments, the second nucleotide from the 5′ end of the sense strand (X2) is a 2′-MOE modified nucleotide with a 154PAT059649-WO-PCT nucleobase T, G, or methylated cytosine (e.g., 5-methylcytosine or N4-methylcytosine). In some embodiments, the second nucleotide from the 5′ end of the sense strand (X2) is a 2′- MOE modified nucleotide with a nucleobase T. In some embodiments, the second nucleotide from the 5′ end of the sense strand (X2) is a 2′-MOE modified nucleotide with a nucleobase G. In some embodiments, the second nucleotide from the 5′ end of the sense strand (X2) is a 2′-MOE modified nucleotide with a nucleobase methylated cytosine (e.g., 5-methylcytosine or N4-methylcytosine).

[0353] In some embodiments, the first nucleotide from the 3′ end of the sense strand (X21) is a 2′-MOE modified nucleotide with a nucleobase A or T. In some embodiments, the first nucleotide from the 3′ end of the sense strand (X21) is a 2′-MOE modified nucleotide with a nucleobase A. In some embodiments, the first nucleotide from the 3′ end of the sense strand (X21) is a 2′-MOE modified nucleotide with a nucleobase T. In some embodiments, the second nucleotide from the 3′ end of the sense strand (X20) is a 2′-MOE modified nucleotide with a nucleobase A.

[0354] In certain aspects, X3to X19do not include a 2′-MOE modified nucleotide. In some embodiments, each X3 to X19 is independently selected from 2′-F modified nucleotides and 2′-OMe modified nucleotides.

[0355] In some embodiments, at least one of X3 to X19 is not a deoxyribonucleotide. In some embodiments, X3to X19does not include a deoxyribonucleotide.

[0356] In some embodiments, each Xf, Xf+2, Xf+3, and Xf+4 is 2′-F modified nucleotide when f is an integer from 3 to 17. In some embodiments, f is 5. In some embodiments, f is 6. In some embodiments, f is 7. In some embodiments, f is 8. In some embodiments, f is 9. In some embodiments, X5, X7, X8, and X9are 2′-F modified nucleotides. In some embodiments, X6, X8, X9, and X10 are 2′-F modified nucleotides. In some embodiments, X7, X9, X10, and X11are 2′-F modified nucleotides. In some embodiments, X8, X10, X11, and X12are 2′-F modified nucleotides. In some embodiments, X9, X11, X12, and X13 are 2′-F modified nucleotides.

[0357] In some embodiments, the sense strand includes 2′-OMe modified nucleotides in the remaining positions in the sense strand.

[0358] In some embodiments, at least two nucleotides from X1, X2, X19, and X20 contain a 3′-PS group, respectively. In some embodiments, two nucleotides from X1, X2, X19, and X20 contain a 3′-PS group, respectively. In some embodiments, three nucleotides from X1, X2, X19, and X20 contain a 3′-PS group, respectively. In some embodiments, each X1, X2, X19, 155PAT059649-WO-PCT and X20 contains a 3′-PS group. In some embodiments, each X1 and X2 contains a 3′-PS group.

[0359] In some embodiments, at least four from X1, X2, X3, X4, X17, X18, X19, and / or X20 contain 3′-PS groups. In some embodiments, four from X1, X2, X3, X4, X17, X18, X19, and / or X20 contain a 3′-PS group, respectively. In some embodiments, six from X1, X2, X3, X4, X17, X18, X19, and / or X20contain a 3′-PS group, respectively. In some embodiments, X1, X2, X3, X4, X17, X18, X19, and X20 contain a 3′-PS group, respectively.

[0360] In some embodiments, in X3to X18, two to six nucleotides contain 3′-PS groups. In some embodiments, in X3 to X18, two nucleotides contain a 3′-PS group, respectively, respectively. In some embodiments, in X3to X18, three nucleotides contain a 3′-PS group, respectively. In some embodiments, in X3 to X18, four nucleotides contain a 3′-PS group, respectively. In some embodiments, in X3 to X18, five nucleotides contain a 3′-PS group, respectively. In some embodiments, in X3 to X18, six nucleotides contain a 3′-PS group, respectively.

[0361] In certain aspects, the sense strand includes 2′-MOE modified nucleotides positioned at the 1st, 2nd, 20th, and 21stnucleotides from the 5′-end of the sense strand.

[0362] In some embodiments, the sense strand having 21 nucleotides in length includes: (i) 2′-MOE modifications at the 1st, 2nd, 20thand / or 21stnucleotides from the 5′-end of the sense strand; and (ii) 3′-PS modifications at the 1st, 2nd, 19th, and / or 20thnucleotides from 5′-end of the sense strand.

[0363] In some embodiments, the sense strand having 21 nucleotides in length includes: (i) 2′-MOE modifications at the 1st, 2nd, 20thand / or 21stnucleotides from the 5′-end of the sense strand; and 2′-F modifications at 8th, 10th, 11th, and 12thnucleotides from the 5′ end of the sense strand.

[0364] In some embodiments, the sense strand having 21 nucleotides in length includes: (i) 2′-MOE modifications at the 1st, 2nd, 20thand / or 21stnucleotides from the 5′-end of the sense strand; 2′-F modifications at 7th, 9th, 10th, and 11thnucleotides from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining nucleotides.

[0365] In some embodiments, the sense strand having 21 nucleotides in length includes: (i) 2′-MOE modifications at the 1st, 2nd, 20thand / or 21stnucleotides from the 5′-end of the sense strand; and 2′-F modifications at 7th, 9th, 10th, and 11thnucleotides from the 5′ end of the sense strand, and 156PAT059649-WO-PCT (ii) 3′-PS modifications at the 1stand 2ndnucleotides from 5′-end of the sense strand.

[0366] In some embodiments, the sense strand having 21 nucleotides in length includes: (i) 2′-MOE modifications at the 1st, 2nd, 20th, and 21stnucleotides from the 5′-end of the sense strand; 2′-F modifications at 7th, 9th, 10th, and / or 11thnucleotides from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining nucleotides, and (ii) 3′-PS modifications at the 1stand 2ndnucleotides from 5′-end of the sense strand.

[0367] In some embodiments, the sense strand having 21 nucleotides in length includes: (i) 2′-MOE modifications at the 1st, 2nd, 20thand / or 21stnucleotides from the 5′-end of the sense strand; and 2′-F modifications at 7th, 9th, 10th, and 11thnucleotides from the 5′ end of the sense strand, and (ii) 3′-PS modifications at the 1st, 2nd, 19th, and 20thnucleotides from 5′ end of the sense strand.

[0368] In some embodiments, the sense strand having 21 nucleotides in length includes: (i) 2′-MOE modifications at the 1st, 2nd, 20th, and 21stnucleotides from the 5′-end of the sense strand; 2′-F modifications at 7th, 9th, 10th, and / or 11thnucleotides from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining nucleotides, and (ii) 3′-PS modifications at the 1st, 2nd, 19th, and 20thnucleotides from 5′ end of the sense strand.

[0369] In certain aspects, a sense strand of the dsRNA as described herein may have a Formula (I′), 5′-Y1′-Y2′-Y3-Y4-Y5-Y6-Y7-Y8-Y9-Y10-Y11-Y12-Y13-Y14-Y15-Y16-Y17-Y18-Y19-Y20-Y21-3′ (I′) wherein: each Y1, Y2,Y20, and Y21is independently a TNA; and each Y3 to Y19 is independently selected from 2′-deoxy modified nucleotide, 2′-MOE modified nucleotide, 2′-F modified nucleotide, and 2′-OMe modified nucleotide.

[0370] In certain aspects, in Formula (I′), Y3 to Y19 do not include a 2′-MOE modified nucleotide. In some embodiments, each Y3to Y19is selected from deoxyribonucleotide, 2′-F modified nucleotides and 2′-OMe modified nucleotides. In some embodiments, at least one of Y3to Y19is not a deoxyribonucleotide.

[0371] In some embodiments, each Yf, Yf+2, and Yf+3 is 2′-F modified nucleotide and Yf+4 is 2′-deoxy modified nucleotide when f is an integer from 3 to 17. In some embodiments, f is 157PAT059649-WO-PCT 5. In some embodiments, f is 6. In some embodiments, f is 7. In some embodiments, f is 8. In some embodiments, f is 9. In some embodiments, Y5, Y7, and Y8are 2′-F modified nucleotides, and Y9 is 2′-deoxy modified nucleotide (e.g., dT). In some embodiments, Y6, Y8, and Y9are 2′-F modified nucleotides and Y10is 2′-deoxy modified nucleotide (e.g., dT). In some embodiments, Y7, Y9, and Y10 are 2′-F modified nucleotides, and Y11 is 2′-deoxy modified nucleotide (e.g., dT). In some embodiments, Y8, Y10, and Y11are 2′-F modified nucleotides, and Y12 is 2′-deoxy modified nucleotide (e.g., dT). In some embodiments, Y9, Y11, and Y12 are 2′-F modified nucleotides, and Y13is 2′-deoxy modified nucleotide (e.g., dT).

[0372] In some embodiments, the sense strand includes 2′-OMe modified nucleotides in the remaining positions in the sense strand.

[0373] In some embodiments, at least two nucleotides from Y1, Y2, Y19, and Y20 contain a 3′-PS group, respectively. In some embodiments, two from Y1, Y2, Y19, and Y20 contains 3′-PS group. In some embodiments, three nucleotides from Y1, Y2, Y19, and Y20 contain a 3′- PS group, respectively. In some embodiments, each Y1, Y2, Y19, and Y20 contains a 3′-PS group. In some embodiments, each Y1and Y2contains a 3′-PS group.

[0374] In some embodiments, at least four from Y1, Y2, Y3, Y4, Y17, Y18, Y19, and / or Y20 contain 3′-PS groups. In some embodiments, four from Y1, Y2, Y3, Y4, Y17, Y18, Y19, and / or Y20 contain a 3′-PS group, respectively. In some embodiments, six from Y1, Y2, Y3, Y4, Y17, Y18, Y19, and / or Y20contain a 3′-PS group, respectively. In some embodiments, Y1, Y2, Y3, Y4, Y17, Y18, Y19, and Y20 contain a 3′-PS group, respectively.

[0375] In some embodiments, in Y3to Y18, two to six nucleotides contain 3′-PS groups. In some embodiments, in Y3 to Y18, two nucleotides contain a 3′-PS group, respectively. In some embodiments, in Y3to Y18, three nucleotides contain a 3′-PS group, respectively. In some embodiments, in Y3 to Y18, four nucleotides contain a 3′-PS group, respectively. In some embodiments, in Y3to Y18, five nucleotides contain a 3′-PS group, respectively. In some embodiments, in Y3 to Y18, six nucleotides contain a 3′-PS group, respectively.

[0376] In certain aspects, the sense strand includes TNAs positioned at the 1st, 2nd, 20th, and 21st nucleotides from the 5′ end of the sense strand.

[0377] In some embodiments, the sense strand having 21 nucleotides in length includes: (i) TNAs at the 1st, 2nd, 20th and / or 21st nucleotides from the 5′ end of the sense strand; and (ii) 3′-PS modifications at the 1st, 2nd, 19th, and / or 20th nucleotides from 5′ end of the sense strand. 158PAT059649-WO-PCT

[0378] In some embodiments, the sense strand having 21 nucleotides in length includes: (i) TNAs at the 1st, 2nd, 20th and / or 21st nucleotides from the 5′ end of the sense strand; 2′-F modifications at 7th, 9th, 10th, and 11th nucleotides from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining nucleotides.

[0379] In some embodiments, the sense strand having 21 nucleotides in length includes: (i) TNAs at the 1st, 2nd, 20th and / or 21st nucleotides from the 5′ end of the sense strand; 2′-F modifications at 7th, 9th, 10th and 11th nucleotides from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining nucleotides; and (ii) 3′-PS modifications at the 1st and 2nd nucleotides from 5′ end of the sense strand.

[0380] In some embodiments, the sense strand having 21 nucleotides in length includes: (i) TNAs at the 1st, 2nd, 20th and / or 21st nucleotides from the 5′ end of the sense strand; 2′-F modifications at 7th, 9th, 10th and 11th nucleotides from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining nucleotides; and (ii) 3′-PS modifications at the 1st, 2nd, 19th, and 20th nucleotides from 5′ end of the sense strand.

[0381] In some embodiments, the sense strand having 21 nucleotides in length includes: (i) TNAs at the 1st, 2nd, 20th and 21st nucleotides from the 5′ end of the sense strand; 2′-F modifications at 7th, 9th, 10th and 11th nucleotides from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining nucleotides; and (ii) 3′-PS modifications at the 1st and 2nd nucleotides from 5′ end of the sense strand.

[0382] In some embodiments, the sense strand having 21 nucleotides in length includes: (i) TNAs at the 1st, 2nd, 20th and 21st nucleotides from the 5′ end of the sense strand; 2′-F modifications at 7th, 9th, 10th and 11th nucleotides from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining nucleotides; and (ii) 3′-PS modifications at the 1st, 2nd, 19th, and 20th nucleotides from 5′ end of the sense strand.

[0383] Example modification patterns of sense strands are shown in Table B. Table B 21-mer SS 2′-MOE TNA 2′F modified 2′-OMe 3′-PS linkage modification modified osition nucleotide modified159PAT059649-WO-PCT 17, 18, 19, 20, 21 SS2 1, 2, 20, 21 7, 9, 10, 11 3, 4, 5, 6, 8, 1, 2 [038, as the modification pattern of SS1. In some embodiments, the sense strand having 21 nucleotides in length does not have the modification pattern of SS1. In some embodiments, the sense strand having 21 nucleotides in length has the modification pattern of SS2. In some embodiments, the sense strand having 21 nucleotides in length has the modification pattern of SS3. In some embodiments, the sense strand having 21 nucleotides in length has the modification pattern of SS4. In some embodiments, the sense strand having 21 nucleotides in length has the modification pattern of SS5. Antisense Strand (AS)

[0385] In certain aspects, an antisense strand of the dsRNA as described herein are substantially (e.g., greater than about 80%, 85%, 90%, or 95% of the total nucleotides) made of modified nucleotides. In another certain aspect, the antisense strand is entirely made of modified nucleotides.

[0386] In certain aspects, the first nucleotide from the 5′ end of the antisense strand may contain an additional phosphate group or a variant thereof (e.g., phosphorothioate, phosphorodithioate, methylphosphonate, methylene phosphonate, or vinylphosphonate (VP)) attached or linked to the 5′ terminal group of the first nucleotide) attached or linked to the 5′ terminal group of the first nucleotide.

[0387] In certain aspects, the antisense strand includes 5′-vinylphosphonate (5′-VP) group at the first nucleotide from the 5′ end in the antisense strand. The “5′-VP” is a 160PAT059649-WO-PCT chemical moiety having the structure of , or a pharmaceutically acceptable salt thereof, where the wavy line represent of attachment to the 5′ carbon of thepentofuranosyl sugar of a nucleotide.

[0388] In some embodiments, the first nucleotide from the 5′ end in the antisense strand includes (E)-vinylphosphonate (VP) having a structure of , or a pharmaceutically acceptable salt thereof, wherein the wavy line presents the point attachment to the 4′carbon of the pentofuranosyl sugar of a nucleotide. In some embodiments, the first nucleotide from the 5′ end in the antisense strand includes (Z)-vinylphosphonate having a structure of , or a pharmaceutically acceptable salt thereof, wherein the wavy line presents the point otachment to the 4′ carbon of the pentofuranosyl sugar of a nucleotide.

[0389] In some embodiments, the first nucleotide from the 5′ end of the antisense strand has a structure lly acceptable salantisense strand has a structur r a pharmaceutically acceptable salt thereof. In some embodim, the 5′ end of the antisense strand 161PAT059649-WO-PCT allyantisense strand has a structure of r a pharmaceutically acceptable salt thereof.

[0390] In some embodiments, the first nucleotide from the 5′ end of the antisense strand has a structure r a pharmaceutically acceptable salttereo, weren s an attacment pont to te adjacent nucleotides. In some embodiments, the first neotide from the 5′ end of the antisense strand has a structure a pharmaceutically acceptable salt thereof. In some, ide from the 5′ end of the antisense strand has a structure of 162PAT059649-WO-PCT a pharmaceutically acceptable salt thereof,nt nucleotides. In some embodiments, the first nu otide from the 5′ end of the antisense strand has a structure of a pharmaceutically acceptable salt thereof.ects, the antisense strand of the dsRNA as described herein includes two or more 2′-F modifications. In some embodiments, the antisense strand of the dsRNA includes two, three, four, five, six, seven, or eight 2′-F modified nucleotides. In some embodiments, the antisense strand includes two 2′-F modified nucleotides. In some embodiments, the antisense strand includes three 2′-F modified nucleotides. In some embodiments, the antisense strand includes four 2′-F modified nucleotides. In some embodiments, the antisense strand includes five 2′-F modified nucleotides. In some embodiments, the antisense strand includes six 2′-F modified nucleotides. In some embodiments, the antisense strand includes seven 2′-F modified nucleotides. In some embodiments, the antisense strand includes eight 2′-F modified nucleotides. In some embodiments, two contiguous 2′-F modified nucleotides locate in the antisense strand. In some embodiments, three contiguous 2′-F modified nucleotides locate in the antisense strand. In some embodiments, four contiguous 2′-F modified nucleotides locate in the antisense strand.

[0392] In certain aspects, the antisense strand is 23 nucleotides in length. In some embodiments, the antisense includes comprises two, three, or four 2′-F modifications positioned at the 2nd, 6th, 14th, and / or 16th nucleotide from 5′ end of the antisense strand. In some embodiments, the antisense includes two 2′-F modifications positioned at the 2nd, 6th, 163PAT059649-WO-PCT 14th, and / or 16th nucleotide from 5′ end of the antisense strand. In some embodiments, the antisense includes three 2′-F modifications positioned at the 2nd, 6th, 14th, and / or 16th nucleotide from 5′ end of the antisense strand. In some embodiments, the antisense includes 2′-F modifications positioned at the 2nd, 6th, 14th, and 16th nucleotide from 5′ end of the antisense strand.

[0393] In certain aspects, an antisense strand of the dsRNA as described herein does not include a 2′-MOE modification. Alternatively, in certain aspects, the antisense strand includes one to four 2′-MOE modified nucleotides. In some embodiments, the antisense strand includes one 2′-MOE modified nucleotide. In some embodiments, the antisense strand includes two 2′-MOE modified nucleotides. In some embodiments, the antisense strand includes three 2′-MOE modified nucleotides. In some embodiments, the antisense strand includes four 2′-MOE modified nucleotides.

[0394] In certain aspects, the antisense strand is 23 nucleotides in length. In some embodiments, the antisense strand includes one to four 2′-MOE modification at the 1st, 9th, 10th, and 23rd nucleotides from the 5′-end of the antisense strand. In some embodiments, the antisense strand includes one 2′-MOE modification at the 1st, 9th, 10th, or 23rd nucleotides from the 5′-end of the antisense strand. In some embodiments, the antisense strand includes two 2′-MOE modifications at the 1st, 9th, 10th, and / or 23rd nucleotides from the 5′-end of the antisense strand. In some embodiments, the antisense strand includes three 2′-MOE modifications at the 1st, 9th, 10th, and / or 23rd nucleotides from the 5′-end of the antisense strand. In some embodiments, the antisense strand includes four 2′-MOE modification at the 1st, 9th, 10th, and 23rd nucleotides from the 5′-end of the antisense strand.

[0395] In certain aspects, the antisense strand includes at least one GNA nucleotide. In some embodiments, the antisense strand includes only one GNA.

[0396] In certain aspects, the antisense strand is 23 nucleotides in length. In some embodiments, the antisense strand includes only one GNA at the 4th nucleotide from 5′-end of the antisense strand. In some embodiments, the antisense strand includes only one GNA at the 5th nucleotide from 5′-end of the antisense strand. In some embodiments, the antisense strand includes only one GNA at the 6th nucleotide from 5′-end of the antisense strand.

[0397] In certain aspects, the antisense strand includes two, three, or four phosphorothioate (PS) linkages between nucleosides. In certain aspects, the antisense strand is 23 nucleotides in length. In some embodiments, the antisense strand includes two 3′-PS modifications positioned at the 1st, 2nd, 21st, and / or 22nd nucleotides from 5′-end of the 164PAT059649-WO-PCT antisense strand. In some embodiments, the antisense strand includes three 3′-PS modifications positioned at the 1st, 2nd, 21st, and / or 22nd nucleotides from 5′-end of the antisense strand. In some embodiments, the antisense strand includes 3′-PS modifications positioned at the 1st, 2nd, 21st, and 22nd nucleotides from 5′-end of the antisense strand.

[0398] In certain aspects, the antisense strand includes two to eight phosphorothioate (PS) linkages between nucleosides.

[0399] In certain aspects, the antisense strand is 23 nucleotides in length. In some embodiments, the antisense strand includes two 3′-PS modified nucleotides positioned at the 1st, 2nd, 3rd, 4th, 19th, 20th, 21st, and / or 22nd nucleotides from 5′-end of the antisense strand. In some embodiments, the antisense strand includes four 3′-PS modified nucleotides positioned at the 1st, 2nd, 3rd, 4th, 19th, 20th, 21st, and / or 22nd nucleotides from 5′-end of the antisense strand. In some embodiments, the antisense strand includes six 3′-PS modified nucleotides positioned at the 1st, 2nd, 3rd, 4th, 19th, 20th, 21st, and / or 22nd nucleotides from 5′-end of the antisense strand. In some embodiments, the antisense strand includes 3′-PS modified nucleotides positioned at the 1st, 2nd, 3rd, 4th, 19th, 20th, 21st, and 22nd nucleotides from 5′-end of the antisense strand.

[0400] In certain aspects, the antisense strand is 23 nucleotides in length. In some embodiments, at least one of the PS groups at the 1st, 2nd, 3rd, 4th, 19th, 20th, 21st, and / or 22nd nucleotides from 5′-end of the antisense strand is a stereopure Rp isomer. In some embodiments, at least one of the PS groups at the 1st, 2nd, 21st, and / or 22nd nucleotides from 5′-end of the antisense strand is a stereopure Rp isomer. In some embodiments, at least one of the PS groups at the 1st and / or 22nd nucleotides from 5′-end of the antisense strand is a stereopure Rp isomer. In some embodiments, the PS group at the 1st nucleotide from 5′-end of the antisense strand is a stereopure Rp isomer. In some embodiments, the PS group at the 22nd nucleotide from 5′-end of the antisense strand is a stereopure Rp isomer. In some embodiments, the PS groups at the 1st and 22nd nucleotides from 5′-end of the antisense strand are stereopure Rp isomers.

[0401] In certain aspects, the antisense strand is 23 nucleotides in length. In some embodiments, at least one of the PS groups at the 1st, 2nd, 3rd, 4th, 19th, 20th, 21st, and / or 22nd nucleotides from 5′-end of the antisense strand is a stereopure Sp isomer. In some embodiments, at least one of the PS groups at the 1st, 2nd, 21st, and / or 22nd nucleotides from 5′-end of the antisense strand is a stereopure Sp isomer. In some embodiments, at least one of the PS groups at the 1st and / or 22nd nucleotides from 5′-end of the antisense strand is a 165PAT059649-WO-PCT stereopure Sp isomer. In some embodiments, the PS group at the 1st nucleotide from 5′-end of the antisense strand is a stereopure Sp isomer. In some embodiments, the PS group at the 22nd nucleotide from 5′-end of the antisense strand is a stereopure Sp isomer. In some embodiments, the PS groups at the 1st and 22nd nucleotides from 5′-end of the antisense strand are stereopure Sp isomers.

[0402] In certain aspects, the antisense strand is 23 nucleotides in length. In some embodiments, at least one of the PS groups at the 1st, 2nd, 3rd, 4th, 19th, 20th, 21st, and / or 22nd nucleotides from 5′-end of the antisense strand is stereopure Sp isomer

[0403] In certain aspects, an antisense strand of dsRNA may have a Formula (II): 5′-X1′-X2′-X3′-X4′-X5′-X6′-X7′-X8′-X9′-X10′-X11′-X12′-X13′-X14′-X15′-X16′-X17′-X18′-X19′-X20′- X21′-X22′-X23′ -3′ (II) wherein: each X1′ to X23′ is independently selected from a 2′-deoxy modified nucleotide (deoxyribonucleotide), 2′-F modified nucleotide, 2′-OMe modified nucleotide, 2′- MOE modified nucleotide, and GNA; and X1′ further includes a 5′-(E)-vinylphosphonate group.

[0404] In certain aspects, X1′to X23′do not include a 2′-MOE modified nucleotide. In some embodiments, each X1′ to X23′ is independently selected from 2′-F modified nucleotides and 2′-OMe modified nucleotides.

[0405] Alternatively, in certain aspect, X1′ to X23′ include one to four 2′-MOE modified nucleotides. In some embodiments, one of X1′, X9′, X10′, and X23′may be 2′-MOE modified nucleotide. In some embodiments, two of X1′, X9′, X10′, and X23′ may be 2′-MOE modified nucleotides. In some embodiments, three of X1′, X9′, X10′, and X23′may be 2′-MOE modified nucleotides. In some embodiments, X1′, X9′, X10′, and X23′ may be 2′-MOE modified nucleotide.

[0406] In some embodiments, X2′ is a 2′-F modified nucleotide. In some embodiments, X6′is a 2′-F modified nucleotide. In some embodiments, X14′is a 2′-F modified nucleotide. In some embodiments, X16′ is a 2′-F modified nucleotide. In some embodiments, two of X2′, X6′, X14′and X16′are 2′-F modified nucleotides. In some embodiments, three of X2′, X6′, X14′and X16′ are 2′-F modified nucleotides. In some embodiments, each X2′, X6′, X14′ and X16′ is a 2′-F modified nucleotide.

[0407] In some embodiments, X1′ to X23′ may include at least one GNA. In some embodiments, X1′ to X23′ may include only one GNA. In some embodiments, X5′ is a GNA. 166PAT059649-WO-PCT

[0408] In some embodiments, the antisense strand includes 2′-OMe modified nucleotides in the remaining positions in the antisense strand.

[0409] In some embodiments, at least two from X1′, X2′, X21′, and X22′ contain 3′-PS groups. In some embodiments, two from X1′, X2′, X21′, and X22′contain a 3′-PS group, respectively. In some embodiments, three from X1′, X2′, X21′, and X22′ contain a 3′-PS group. In some embodiments, each X1′, X2′, X21′, and X22′contains a 3′-PS group.

[0410] In some embodiments, at least four from X1′, X2′, X3′, X4′, X19′, X20′, X21′, and X22′ contain 3′-PS groups. In some embodiments, four from X1′, X2′, X3′, X4′, X19′, X20′, X21′, and X22′ contain a 3′-PS group, respectively. In some embodiments, six from X1′, X2′, X3′, X4′, X19′, X20′, X21′, and X22′contain a 3′-PS group, respectively. In some embodiments, X1′, X2′, X3′, X4′, X19′, X20′, X21′, and X22′ contain a 3′-PS group, respectively.

[0411] In some embodiments, in X3′ to X20′, two to six nucleotides contain 3′-PS groups. In some embodiments, in X3′ to X20′, two nucleotides contain a 3′-PS group, respectively. In some embodiments, in X3′ to X20′, three nucleotides contain a 3′-PS group, respectively. In some embodiments, in X3′to X20′, four nucleotides contain a 3′-PS group, respectively. In some embodiments, in X3′ to X20′, five nucleotides contain a 3′-PS group, respectively. In some embodiments, in X3′to X20′, six nucleotides contain a 3′-PS group, respectively.

[0412] In certain aspects, the antisense strand includes 5′-(E)-VP modified nucleotide at the first nucleotide from 5′ end of the antisense strand. In some embodiments, the antisense strand includes a 5′-(E)-VP-2′-OMe modified nucleotide at the first nucleotide from 5′ end of the antisense strand.

[0413] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; and (ii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′-end of the antisense strand.

[0414] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; and (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand. 167PAT059649-WO-PCT

[0415] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand, and (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides.

[0416] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′-end of the antisense strand.

[0417] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modification at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modification in the remaining nucleotides, and (iii) 3′-PS modification at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′-end of the antisense strand.

[0418] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modification at 2nd, 6th, 14th, and 16thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modification in the remaining nucleotides, and (iii) 3′-PS modification at the 1st, 2nd, 21st, and 22ndnucleotides from 5′-end of the antisense strand.

[0419] In some embodiments, the antisense strand having 23 nucleotides in length includes: 168PAT059649-WO-PCT (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; GNA at 5thnucleotide from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0420] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; GNA at 6thnucleotide from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0421] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; GNA at 7thnucleotide from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0422] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; TNA at 3rdnucleotide from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and 169PAT059649-WO-PCT (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0423] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; TNA at 5thnucleotide from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0424] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; TNA at 6thnucleotide from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0425] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; TNA at 7thnucleotide from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0426] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; 170PAT059649-WO-PCT (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; a 2′-deoxy modification at 5thnucleotide from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0427] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; a 2′-deoxy modification at 6thnucleotide from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0428] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; a 2′-deoxy modification at 7thnucleotide from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0429] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; GNA at 3rdand 5thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand. 171PAT059649-WO-PCT

[0430] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; GNA at 3rdand 6thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0431] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16th nucleotides from the 5′ end of the antisense strand; GNA at 3rd and 7thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0432] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; TNA at 3rdand 5thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0433] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; 172PAT059649-WO-PCT (ii) 2′-F modifications at 2nd, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; TNA at 3rdand 6thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0434] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; TNA at 3rdand 7thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0435] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; a 2′-deoxy modification at 3rdand 5thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand.

[0436] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; a 2′-deoxy modification at 3rdand 6thnucleotide from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand. 173PAT059649-WO-PCT

[0437] In some embodiments, the antisense strand having 23 nucleotides in length includes: (i) a 5′-(E)-VP-2′-OMe modification at the first nucleotide from 5′ end of the antisense strand; (ii) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; a 2′-deoxy modification at 3rdand 7thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides; and (iii) 3′-PS modifications at the 1st, 2nd, 21st, and / or 22ndnucleotides from 5′ end of the antisense strand. Table C 23-mer AS 5′-VP 2′-F modified GNA TNA 2′-deoxy 2′-OMe 3′-PS modification modified nucleotide position position modified modified linkage tt rn n l tid n l tid n l tid 2 2 2 2 2 2 2 2174PAT059649-WO-PCT 23-mer AS 5′-VP 2′-F modified GNA TNA 2′-deoxy 2′-OMe 3′-PS modification modified nucleotide position position modified modified linkage pattern nucleotide nucleotide nucleotide 2 2 2 2 2 2 2 2 2 2 2 2 2175PAT059649-WO-PCT

[0438] In an aspect, the dsRNAi agent having the nucleotides modification patterns as described herein can improve half-life relative to a reference dsRNAi agent that does not contain such nucleotides modification patterns. In some embodiments, the dsRNAi agent having the nucleotides modification patterns as described herein improved half-life by about 1.1 fold, 1.2 fold, 1.3 fold, 1.4 fold, 1.5 fold, 1.6 fold, 1.7 fold, 1.8 fold, 1.9 fold, 2.0 fold, 3.0 fold, 4.0 fold, 5.0 fold, 6.0 fold, 7.0 fold, 8.0 fold, 9.0 fold, 10 fold, or more relative to a reference dsRNAi agent that does not contain such nucleotides modification patterns. In some embodiments, the dsRNAi agent having the nucleotides modification patterns as described herein improved half-life by about 2 fold, 2.5 fold, 3 fold, 3.5 fold, 4 fold, 4.5 fold, 5 fold, 7 fold, or 10 fold, relative to a reference dsRNAi agent that does not contain such nucleotides modification patterns. dsRNA modification pattern

[0439] In an aspect, the dsRNA as described herein includes a sense strand of Formula (I) as described herein and an antisense strand of Formula (II) as described herein. The sense strand and the antisense strand form a duplex.

[0440] In certain aspects, the dsRNA includes: (i) a sense strand including 2′-MOE modifications at X1, X2, X20 and X21; and (ii) an antisense strand including a 5′-(E)-VP-2′-OMe modification at X1′.

[0441] In some embodiments, the dsRNA includes: (i) a sense strand including: (a) 2′-MOE modifications at X1, X2, X20 and X21; and (b) 3′-PS modifications at X1and X2,and (ii) an antisense strand including: (a) a 5′-(E)-VP-2′-OMe modification at X1′; and (b) 3′-PS modifications at X1′, X2′, X21′, and X22′.

[0442] In some embodiments, the dsRNA includes: (i) a sense strand including: (a) 2′-MOE modifications at X1, X2, X20and X21; and 2′-F modifications at X7, X9, X10 and X11, and (ii) an antisense strand including: (a) a 5′-(E)-VP-2′-OMe modification at X1′, and (b) 2′-F modifications at X2′, X6′, X14′ and / or X16′. 176PAT059649-WO-PCT

[0443] In some embodiments, the dsRNA includes: (i) a sense strand including: (a) 2′-MOE modifications at X1, X2, X20 and X21; and 2′-F modifications at X7, X9, X10and X11, and (b) 3′-PS modifications at X1 and X2, and (ii) an antisense strand including: (a) a 5′-(E)-VP-2′-OMe modification at X1′; (b) 2′-F modifications at X2′, X6′, X14′and / or X16′; and (c) 3′-PS modifications X1′, X2′, X21′, and X22′.

[0444] In some embodiments, the dsRNA includes: (i) a sense strand including: (a) 2′-MOE modifications at X1, X2, X20 and X21; 2′-F modifications at X7, X9, X10 and X11; and 2′-OMe modifications in the remaining nucleotides in the sense strand, and (b) 3′-PS modifications at X1and X2, and (ii) an antisense strand including: (a) a 5′-(E)-VP-2′-OMe modification at X1′, (b) 2′-F modifications at X2′, X6′, X14′ and / or X16′; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, and (c) 3′-PS modifications at X1′, X2′, X21′, and X22′.

[0445] In some embodiments, the dsRNA includes: (i) a sense strand including: (a) 2′-MOE modifications at X1, X2, X20and X21; 2′-F modifications at X7, X9, X10and X11; and 2′-OMe modifications in the remaining nucleotides in the sense strand, and (b) 3′-PS modifications at X1 and X2, and (ii) an antisense strand including: (a) a 5′-(E)-VP-2′-OMe modification at X1′, (b) a GNA nucleotide at X5′, (c) 2′-F modifications at X2′, X6′, X14′ and / or X16′; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, and (d) 3′-PS modifications at X1′, X2′, X21′, and X22′.

[0446] In some embodiments, the dsRNA includes: 177PAT059649-WO-PCT (i) a sense strand including: (a) 2′-MOE modifications at X1, X2, X20and X21; 2′-F modifications at X7, X9, X10and X11; and 2′-OMe modifications in the remaining nucleotides in the sense strand, and (b) 3′-PS modifications at X1 and X2, and (ii) an antisense strand including: (a) a 5′-(E)-VP-2′-OMe modification at X1′; (b) 3′-PS modifications at X1′, X2′, X21′, and X22′; and (c) one selected from: (c1) 2′-F modifications at X2′, X6′, X14′and X16′; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, (c2) 2′-F modifications at X2′, X6′, X14′ and X16′; TNA at X3′; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, (c3) 2′-F modifications at X2′, X6′, X14′ and X16′; TNA, GNA or 2′-deoxy modification at X5′; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, (c4) 2′-F modifications at X2′, X14′and X16′; TNA, GNA or 2′-deoxy modification at X6′; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, and (c5) 2′-F modifications at X2′, X6′, X14′ and X16′; TNA, GNA or 2′-deoxy modification at X7′; and 2′-OMe modifications in the remaining nucleotides in the antisense strand.

[0447] In some embodiments, the dsRNA includes: (i) a sense strand of Formula (I′) including: (a) TNAs at Y1, Y2, Y20and Y21; and (b) 3′-PS modifications at Y1, Y2, Y19, and Y20, and (ii) an antisense strand of Formula (II) including: (a) a 5′-(E)-VP-2′-OMe modification at X1′; and (b) 3′-PS modifications at X1′, X2′, X21′, and X22′.

[0448] In some embodiments, the dsRNA includes: (i) a sense strand of Formula (I′) including: (a) TNAs at Y1, Y2, Y20 and Y21; 178PAT059649-WO-PCT (b) 3′-PS modifications at Y1, Y2, Y19, and / or Y20; and (c) 2′-F modifications at Y7, Y9, and Y10; 2′-deoYy modification at Y11; and 2′-OMe modifications in the remaining nucleotides in the sense strand, and (ii) an antisense strand of Formula (II) including: (a) a 5′-(E)-VP-2′-OMe modification at X1′; (b) 3′-PS modifications at X1′, X2′, X21′, and X22′; and (c) one selected from: (c1) 2′-F modifications at X2′, X6′, X14′ and X16′; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, (c2) 2′-F modifications at X2′, X6′, X14′ and X16′; TNA at X3′; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, (c3) 2′-F modifications at X2′, X6′, X14′ and X16′; TNA, GNA or 2′-deoxy modification at X5′; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, (c4) 2′-F modifications at X2′, X14′ and X16′; TNA, GNA or 2′-deoxy modification at X6′; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, and (c5) 2′-F modifications at X2′, X6′, X14′and X16′; TNA, GNA or 2′-deoxy modification at X7′; and 2′-OMe modifications in the remaining nucleotides in the antisense strand.

[0449] In certain aspects, the dsRNA includes: (i) a sense strand having 21 nucleotides in length and including 2′-MOE modifications at the 1st, 2nd, 20thand 21stnucleotides from the 5′end of the sense strand; and (ii) an antisense strand having 23 nucleotides in length and including a 5′-(E)-VP-2′- OMe modification at the 1stnucleotide from 5′ end of the antisense strand.

[0450] In some embodiments, the dsRNA as described herein includes a sense strand having 21 nucleotides in length and an antisense strand having 23 nucleotides.

[0451] In some embodiments, the dsRNA as described herein includes a sense strand having 21 nucleotides in length and including 2′-MOE modifications at the 1st, 2nd, 20thand 21stnucleotides from the 5′end of the sense strand. 179PAT059649-WO-PCT

[0452] In some embodiments, the dsRNA as described herein includes antisense strand having 23 nucleotides in length and including a 5′-(E)-VP-2′-OMe modification at the 1stposition from 5′ end of the antisense strand.

[0453] In some embodiments, the dsRNA has Modification Pattern A of: (i) a sense strand having 21 nucleotides in length and including: (a) 2′-MOE modifications at the 1st, 2nd, 20thand 21stnucleotides from the 5′end of the sense strand, and (b) 3′-PS modifications at the 1stand 2ndnucleotides from 5′-end of the sense strand, and (ii) an antisense strand having 23 nucleotides in length and including: (a) a 5′-(E)-VP-2′-OMe modification at the 1stposition from 5′ end of the antisense strand, and (b) 3′-PS modifications at the 1st, 2nd, 21st, and 22ndnucleotides from 5′-end of the antisense strand.

[0454] In some embodiments, the dsRNA has Modification Pattern B of: (i) a sense strand having 21 nucleotides in length and including: (a) 2′-MOE modifications at the 1st, 2nd, 20thand 21stnucleotides from the 5′-end of the sense strand; and 2′-F modifications at the 7th, 9th, 10th, and 11thnucleotides from the 5′ end of the sense strand, and (ii) an antisense strand having 23 nucleotides in length and including: (a) a 5′-(E)-VP-2′-OMe modification at the 1stposition from 5′ end of the antisense strand, and (b) 2′-F modifications at the 2nd, 6th, 14th, and 16thnucleotides from the 5′ end of the antisense strand.

[0455] In some embodiments, the dsRNA has Modification Pattern C of: (i) a sense strand having 21 nucleotides in length and including: (a) 2′-MOE modifications at the 1st, 2nd, 20thand 21stnucleotides from the 5′-end of the sense strand; and 2′-F modifications at the 7th, 9, 10th, and 11thnucleotides from the 5′ end of the sense strand, and (b) 3′-PS modifications at the 1stand 2ndnucleotides from 5′-end of the sense strand, and (ii) an antisense strand having 23 nucleotides in length and including: 180PAT059649-WO-PCT (a) a 5′-(E)-VP-2′-OMe modification at the 1stposition from 5′ end of the antisense strand, (b) 2′-F modifications at the 2nd, 6th, 14th, and 16thnucleotides from the 5′ end of the antisense strand, and (c) 3′-PS modifications at the 1st, 2nd, 21st, and 22ndnucleotides from 5′-end of the antisense strand.

[0456] In some embodiments, the dsRNA has Modification Pattern D of: (i) a sense strand having 21 nucleotides in length and including: (a) 2′-MOE modifications at the 1st, 2nd, 20thand 21stnucleotides from the 5′-end of the sense strand; 2′-F modifications at 7th, 9th, 10th, and 11thnucleotides from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining nucleotides in the sense strand, and (b) 3′-PS modifications at the 1stand 2ndnucleotides from 5′-end of the sense strand, and (ii) an antisense strand having 23 nucleotides in length and including: (a) a 5′-(E)-VP-2′-OMe modification at the 1stposition from 5′ end of the antisense strand, (b) 2′-F modifications at 2nd, 6th, 14th, and 16thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides in the sense strand, and (c) 3′-PS modifications at the 1st, 2nd, 21st, and 22ndnucleotides from 5′-end of the antisense strand.

[0457] In some embodiments, the dsRNA has Modification Pattern E of: (i) a sense strand having 21 monomers (e.g., nucleotides) in length and including: (a) 2′-MOE modifications at the 1st, 2nd, 20thand 21stpositions from the 5′-end of the sense strand; 2′-F modifications at 7th, 9th, 10th, and 11thpositions from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining positions in the sense strand, and (b) 3′-PS modifications at the 1stand 2ndpositions from 5′-end of the sense strand; and (ii) an antisense strand having 23 monomers (e.g., nucleotides) in length and including: (a) a 5′-(E)-VP-2′-OMe modification at the 1stposition from 5′ end of the antisense strand, 181PAT059649-WO-PCT (b) a GNA at 5thposition from the 5′ end of the antisense strand; 2′-F modifications at 2nd, 6th, 14th, and 16thpositions from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining positions in the antisense strand, and (c) 3′-PS modifications at the 1st, 2nd, 21st, and 22nd positions from 5′-end of the antisense strand.

[0458] In some embodiments, the dsRNA has Modification Pattern E-1 of: (i) a sense strand having 21 monomers (e.g., nucleotides) in length and including: (a) 2′-MOE modifications at the 1st, 2nd, 20thand 21stpositions from the 5′-end of the sense strand; 2′-F modifications at 7th, 9th, 10th, and 11thpositions from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining positions in the sense strand, and (b) 3′-PS modifications at the 1stand 2ndpositions from 5′-end of the sense strand; and (ii) an antisense strand having 23 monomers (e.g., nucleotides) in length and including: (a) a 5′-(E)-VP-2′-OMe modification at the 1stposition from 5′ end of the antisense strand, (b) a GNA, TNA, or 2′-deoxy modification at 5thposition from the 5′ end of the antisense strand; 2′-F modifications at 2nd, 6th, 14th, and 16thpositions from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining positions in the antisense strand, and (c) 3′-PS modifications at the 1st, 2nd, 21st, and 22nd positions from 5′-end of the antisense strand.

[0459] In some embodiments, the dsRNA has Modification Pattern E-2 of: (i) a sense strand having 21 monomers (e.g., nucleotides) in length and including: (a) 2′-MOE modifications at the 1st, 2nd, 20thand 21stpositions from the 5′-end of the sense strand; 2′-F modifications at 7th, 9th, 10th, and 11thpositions from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining positions in the sense strand, and (b) 3′-PS modifications at the 1stand 2ndpositions from 5′-end of the sense strand; and (ii) an antisense strand having 23 monomers (e.g., nucleotides) in length and including: 182PAT059649-WO-PCT (a) a 5′-(E)-VP-2′-OMe modification at the 1stposition from 5′ end of the antisense strand, (b) a TNA nucleotide at 3rdposition from the 5′ end of the antisense strand; 2′-F modifications at 2nd, 6th, 14th, and 16thpositions from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining positions in the antisense strand, and (c) 3′-PS modifications at the 1st, 2nd, 21st, and 22nd positions from 5′-end of the antisense strand.

[0460] In some embodiments, the dsRNA has Modification Pattern E-3 of: (i) a sense strand having 21 monomers (e.g., nucleotides) in length and including: (a) 2′-MOE modifications at the 1st, 2nd, 20thand 21stpositions from the 5′-end of the sense strand; 2′-F modifications at 7th, 9th, 10th, and 11thpositions from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining positions in the sense strand, and (b) 3′-PS modifications at the 1stand 2ndpositions from 5′-end of the sense strand; and (ii) an antisense strand having 23 monomers (e.g., nucleotides) in length and including: (a) a 5′-(E)-VP-2′-OMe modification at the 1stposition from 5′ end of the antisense strand, (b) a GNA, TNA, or 2′-deoxy modification at 6thposition from the 5′ end of the antisense strand; 2′-F modifications at 2nd, 14th, and 16thpositions from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining positions in the antisense strand, and (c) 3′-PS modifications at the 1st, 2nd, 21st, and 22nd positions from 5′-end of the antisense strand.

[0461] In some embodiments, the dsRNA has Modification Pattern E-4 of: (i) a sense strand having 21 monomers (e.g., nucleotides) in length and including: (a) 2′-MOE modifications at the 1st, 2nd, 20thand 21stpositions from the 5′-end of the sense strand; 2′-F modifications at 7th, 9th, 10th, and 11thpositions from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining positions in the sense strand, and (b) 3′-PS modifications at the 1stand 2ndpositions from 5′-end of the sense strand; and 183PAT059649-WO-PCT (ii) an antisense strand having 23 monomers (e.g., nucleotides) in length and including: (a) a 5′-(E)-VP-2′-OMe modification at the 1stposition from 5′ end of the antisense strand, (b) a GNA, TNA, or 2′-deoxy modification at 7thposition from the 5′ end of the antisense strand; 2′-F modifications at 2nd, 6th, 14th, and 16thpositions from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining positions in the antisense strand, and (c) 3′-PS modifications at the 1st, 2nd, 21st, and 22nd positions from 5′-end of the antisense strand.

[0462] In certain aspects, the antisense strand of the dsRNA as described herein does not include a 5′-(E)-VP-2′-OMe modification at the 1stposition from 5′ end of the antisense strand.

[0463] In some embodiments, the dsRNA has Modification Pattern F of: (i) a sense strand having 21 nucleotides in length and including: (a) 2′-MOE modifications at the 1st, 2nd, 20thand 21stnucleotides from the 5′-end of the sense strand; 2′-F modifications at 7th, 9th, 10th, and 11thnucleotides from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining nucleotides in the sense strand, and (b) 3′-PS modifications at the 1stand 2ndnucleotides from 5′-end of the sense strand. (ii) an antisense strand having 23 nucleotides in length and including: (a) 2′-F modifications at 2nd, 6th, 14th, and 16thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, and (b) 3′-PS modifications at the 1st, 2nd, 21st, and 22ndnucleotides from 5′-end of the antisense strand.

[0464] In some embodiments, the dsRNA has Modification Pattern G-1 of: (i) a sense strand having 21 nucleotides in length and including: (a) 2′-F modifications at 7th, 9th, 10th, and 11thnucleotides from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining nucleotides in the sense strand, and (b) 3′-PS modifications at the 1stand 2ndnucleotides from 5′-end of the sense strand, and (ii) an antisense strand having 23 nucleotides in length and including: 184PAT059649-WO-PCT (a) 2′-F modifications at 2nd, 6th, 14th, and / or 16thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, and (b) 3′-PS modifications at the 1st, 2nd, 21st, and 22ndnucleotides from 5′-end of the antisense strand.

[0465] In some embodiments, the dsRNA has Modification Pattern G-2 of: (i) a sense strand having 21 nucleotides in length and including: (a) 2′-F modifications at 7th, 9th, 10th, and 11thnucleotides from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining nucleotides in the sense strand, and (b) 3′-PS modifications at the 1stand 2ndnucleotides from 5′-end of the sense strand, and (ii) an antisense strand having 23 nucleotides in length and including: (a) 2′-F modifications at 2nd, 6th, 9th, 14th, and 16thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, and (b) 3′-PS modifications at the 1st, 2nd, 21st, and 22ndnucleotides from 5′-end of the antisense strand.

[0466] In some embodiments, the dsRNA has Modification Pattern H of: (i) a sense strand having 21 nucleotides in length and including: (a) 2′-F modifications at 7th, 9th, 10th, and 11thnucleotides from the 5′ end of the sense strand; and 2′-OMe modifications in the remaining nucleotides in the sense strand, and (b) 3′-PS modifications at the 1stand 2ndnucleotides from 5′-end of the sense strand, and (ii) an antisense strand having 23 nucleotides in length and including: (a) a 5′-(E)-VP-2′-OMe modification at the 1stposition from 5′ end of the antisense strand; (b) 2′-F modifications at 2nd, 6th, 14th, and 16thnucleotides from the 5′ end of the antisense strand; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, and (c) 3′-PS modifications at the 1st, 2nd, 21st, and 22ndnucleotides from 5′-end of the antisense strand. 185PAT059649-WO-PCT

[0467] In some embodiments, the dsRNA has Modification Pattern I of: (i) a sense strand having 21 nucleotides in length and including: (a) TNAs at the 1st, 2nd, 20th and 21st nucleotides from the 5′ end, and (b) 3′-PS modifications at the 1st and 2nd nucleotides from the 5′ end, and (ii) an antisense strand having 23 nucleotides in length and including: (a) a 5′-(E)-VP-2′-OMe modification at the 1st position from the 5′ end, and (b) 3′-PS modifications at the 1st, 2nd, 21st, and 22nd nucleotides from the 5′ end.

[0468] In some embodiments, the dsRNA has Modification Pattern J of: (i) a sense strand having 21 nucleotides in length and including: (a) TNAs at the 1st, 2nd, 20th and 21st nucleotides from the 5′ end, and (b) 3′-PS modifications at the 1st, 2nd, 19th, and 20th nucleotides from the 5′ end, and (ii) an antisense strand having 23 nucleotides in length and including: (a) a 5′-(E)-VP-2′-OMe modification at the 1stposition from the 5′ end, and (b) 3′-PS modifications at the 1st, 2nd, 21st, and 22nd nucleotides from the 5′ end.

[0469] In some embodiments, the dsRNA has Modification Pattern K of: (i) a sense strand having 21 nucleotides in length and including: (a) TNAs at the 1st, 2nd, 20th and 21st nucleotides from the 5′ end, (b) 3′-PS modifications at the 1st, 2nd, 19th, and / or 20th nucleotides from the 5′ end, and (c) 2′-F modifications at the 7th, 9th, 10th, and 11th nucleotides from the 5′ end; and 2′-OMe modifications in the remaining nucleotides in the sense strand, and (ii) an antisense strand having 23 nucleotides in length and including: (a) a 5′-(E)-VP-2′-OMe modification at the 1stposition from the 5′ end, (b) 3′-PS modifications at the 1st, 2nd, 21st, and 22nd nucleotides from the 5′ end, and (c) one selected from: (c1) 2′-F modifications at the 2nd, 6th, 14th, and 16th nucleotides from the 5′ end; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, 186PAT059649-WO-PCT (c2) 2′-F modifications at the 2nd, 6th, 14th, and 16th nucleotides from the 5′ end; TNA at the 3rd nucleotide from the 5′ end; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, (c3) 2′-F modifications at the 2nd, 6th, 14th, and 16th nucleotides from the 5′ end; TNA, GNA or 2′-deoxy modification at the 5th nucleotide from the 5′ end; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, (c4) 2′-F modifications at the 2nd, 14th, and 16th nucleotides from the 5′ end; TNA, GNA or 2′-deoxy modification at the 6th nucleotide from the 5′ end; and 2′-OMe modifications in the remaining nucleotides in the antisense strand, and (c5) 2′-F modifications at the 2nd, 6th, 14th, and 16th nucleotides from the 5′ end; TNA, GNA or 2′-deoxy modification at the 7th nucleotide from the 5′ end; and 2′-OMe modifications in the remaining nucleotides in the antisense strand.

[0470] In certain aspects, modified sequences of sense strands and antisense strands targeting the above indicated HMGCR mRNA (SEQ ID NO: 811, or GenBank: NM_000859.3) are in Table 3. Table 3 siRNA Sequence (5′-3′) Strand SEQ ID187PAT059649-WO-PCT 650T005p001T005p001G004pC004pA004pG004pA007pU004pG007SS 1297 pC007pU007pA004pG004pG004pU004pG004pU004pU004pC004 A005T005188PAT059649-WO-PCT X033A1027p001C007p001A004pG004pA004pU007pU004pU004 AS 1470 pU004pA004pU004pG004pU004pU007pA004pG007pU004pC004 C004U004U004001U004001A004189PAT059649-WO-PCT 726U042p001U042p001G004pC004pA004pG004pA007pU004pG007SS 1482 pC007pU007pA004pG004pG004pU004pG004pU004pU004pC004 A042001A042

[0471] In Table 3, the nucleotide indicated with T is referred to a ribonucleotide having thymidine nucleobase (“ribothymidine”) and “U” is referred to a ribonucleotide having uracil nucleobase (“uridine”). In some embodiments, “T” ribonucleotide (ribothymidine) above may be interchangeably use as “methylated uridine,” “5- methyluridine” or “mU.” 190PAT059649-WO-PCT

[0472] In some embodiments, the sequence in Table 3 may include modified nucleobases. In some embodiments, the sense strand may include one or more nucleotides containing thymin...

Claims

PAT059649-WO-PCT WHAT IS CLAIMED:

1. A double stranded RNAi (dsRNAi) agent comprising: a sense strand comprising a nucleotide sequence selected from SEQ ID NOs: 1 to 405 and 1434 to 1440 in Table 1; and an antisense strand forming a duplex with the sense strand and comprising a nucleotide sequence selected from SEQ ID NOs: 406 to 810 and 1441 to 1447 in Table 1.

2. The dsRNAi agent of claim 1, wherein the sense strand is 21 to 23 nucleotides in length and the antisense strand is 23 to 25 nucleotides in length.

3. The dsRNAi agent of claim 1 or 2, wherein all the nucleotides in the sense strand and the antisense strand are modified nucleotides.

4. The dsRNAi agent of claim 1 through 3, wherein each of the modified nucleotides independently comprises one or more modifications selected from a 2′-deoxy modification, a 2′- O-alkyl modification, a 2′-halo modification, a threofuranosyl nucleotide (TNA) modification, a 2′-5′-linkage modification, a conformationally restricting modification, an abasic modification, a 2′- amino-modification, a 2′-O-allyl modification, 2′-C-alkyl modification, a 2′-O-alkoxyalkyl modification, a morpholino modification, a phosphoramidate modification, a non-natural nucleobase modification, a modification in a tetrahydropyran, a modification containing a 1,5- anhydrohexitol, a modification containing a cyclohexenyl, a modification containing a phosphorothioate group, a modification containing a 5′-vinyl-phosphonate, a modification containing a 5′-phosphate, a modification to form a thermally destabilizing nucleotide, a glycol nucleic acid (GNA) modification, and a 2-O-(N-methylacetamide) modification.

5. The dsRNAi agent of claim 4, wherein each of the modified nucleotides independently comprises one or more modifications selected from 2′-deoxy modification, 2′-O-alkoxyalkyl modification, 2′-O-alkyl modification, 2′-O-allyl modification, 2′-C-allyl modification, 2′-halo 512PAT059649-WO-PCT modification, modification containing a non-natural nucleobase, GNA modification, and TNA modification.

6. The dsRNAi agent of any one of claims 3 through 5, wherein all the modified nucleotides comprise a modification on a 2′ sugar ring.

7. The dsRNAi agent of claim 6, wherein the modified nucleotides are selected from a 2′-O-alkyl modified nucleotide, a 2′-halo modified nucleotide, a 2′-deoxy modified nucleotide, a 2′-O-alkoxyalkyl modified nucleotide and TNA modification.

8. The dsRNAi agent of any one of claims 3 to 7, wherein one or more of the modified nucleotides further comprises a 3′-phosphorothioate (PS) modification.

9. The dsRNAi agent of any one of claims 4 through 8, wherein each of the modified nucleotides independently comprises one or more modifications selected from 2′-deoxy modification, 2′-O-methyl (2′-OMe) modification, 2′-fluoro (2′-F) modification, 2′-O- methoxyethyl (2′-MOE) modification, the modification containing a non-natural nucleobase, TNA, GNA, 3′-phosphorothioate (PS) modification, and 5′-vinyl-phosphonate (5′-VP) modification.

10. The dsRNAi agent of any one of claims 1 to 9, wherein the sense strand comprises one or two 2′-MOE modified nucleotides positioned at the 1stand / or 2ndnucleotides from the 5′-end of the sense strand.

11. The dsRNAi agent of any one of claims 1 to 10, wherein the sense strand comprises one or two 2′-MOE modified nucleotides positioned at the 1stand / or 2ndnucleotides from the 3′- end of the sense strand.

12. The dsRNAi agent of any one of claims 1 to 9, wherein the sense strand comprises one or two TNAs positioned at the 1stand / or 2ndnucleotides from the 5′-end of the sense strand. 513PAT059649-WO-PCT 13. The dsRNAi agent of any one of claims 1 to 10, wherein the sense strand comprises one or two TNAs positioned at the 1stand / or 2ndnucleotides from the 3′-end of the sense strand.

14. The dsRNAi agent of any one of claims 1 through 13, wherein the antisense strand comprises a 5′-VP group at the 1stnucleotide from 5′ end of the antisense strand.

15. The dsRNAi agent of any one of claims 1 through 13, wherein the antisense strand comprises a 5′-(E)-VP group at the 1stnucleotide from 5′ end of the antisense strand.

16. The dsRNAi agent of any one of claims 1 through 13, wherein the antisense strand comprises a 5′-(E)-VP-2′-OMe nucleotide at the 1stposition from 5′ end of the antisense strand.

17. The dsRNAi agent of any one of claims 1 and 16, wherein each of the sense strand and the antisense strand independently comprises two, three, four, five or six 2′-F modified nucleotides.

18. The dsRNAi agent of any one of claims 1 through 17, wherein the sense strand comprises one or two 3′-PS group at the 1stand / or 2ndnucleotides from 5′-end of the sense strand.

19. The dsRNAi agent of any one of claims 1 through 18, wherein the antisense strand comprises one or two 3′-PS group at the 1stand / or 2ndnucleotides from 5′-end of the antisense strand, and / or one or two 3′-PS group at the 1stand / or 2ndnucleotides from 3′-end of the antisense strand.

20. The dsRNAi agent of any one of claims 1 through 19, wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.

21. The dsRNAi agent of claim 20, wherein the sense strand comprises one to four 2′- MOE modified nucleotides positioned at the 1st, 2nd, 20th, and / or 21stnucleotides from the 5′-end of the sense strand. 514PAT059649-WO-PCT 22. The dsRNAi agent of claim 21, wherein the sense strand comprises only four 2′-MOE modified nucleotides.

23. The dsRNAi agent of any one of claims 21 through 22, wherein the sense strand does not comprise a 2′-MOE modified nucleotide at the 3rdto 19thpositions from 5′-end of the sense strand.

24. The dsRNAi agent of claim 20, wherein the sense strand comprises one to four TNAs positioned at the 1st, 2nd, 20th, and / or 21stnucleotides from the 5′-end of the sense strand.

25. The dsRNAi agent of any one of claims 20 through 24, wherein the sense strand comprises two, three, or four 2′-F modified nucleotides positioned at the 7th, 9th, 10th, and / or 11thnucleotide from 5′-end of the sense strand.

26. The dsRNAi agent of claim 25, wherein the sense strand comprises 2′-F modified nucleotides positioned at the 7th, 9th, 10th, and 11thnucleotides from 5′-end of the sense strand.

27. The dsRNAi agent of claim 25 or 26, wherein the remaining nucleotides in the sense strand comprise 2′-OMe modified modification.

28. The dsRNAi agent of any one of claims 20 through 27, wherein the antisense strand comprises a 5′-(E)-VP group at the 1stnucleotide from 5′ end of the antisense strand.

29. The dsRNAi agent of any one of claims 18 through 25, wherein the antisense strand comprises two, three, or four 2′-F modified nucleotides positioned at the 2nd, 6th, 14th, and / or 16thnucleotides from 5′-end of the antisense strand.

30. The dsRNAi agent of claim 29, wherein the antisense strand comprises 2′-F modified nucleotides positioned at the 2nd, 6th, 14th, and 16thnucleotides from 5′-end of the antisense strand. 515PAT059649-WO-PCT 31. The dsRNAi agent of any one of claims 20 through 30, wherein the antisense strand comprises 2′-F modifications positioned at the 2nd, 6th, 14th, and 16th nucleotides from the 5′ end; and (i) a GNA positioned at the 5thnucleotide from 5′ end, or (ii) a TNA positioned at the 3rd nucleotide from the 5′ end.

32. The dsRNAi agent of any one of claims 28 through 31, wherein the remaining nucleotides in antisense strand comprise 2′-OMe modified modifications.

33. The dsRNAi agent of any one of claims 20 through 32, wherein the sense strand comprises one to eight 3′-PS group at the 1st, 2nd, 3rd, 4th, 17th, 18th, 19thand / or 20thnucleotides from 5′-end of the sense strand.

34. The dsRNAi agent of any one of claims 20 through 33, wherein the antisense strand comprises one to eight 3′-PS group at the 1st, 2nd, 3rd, 4th, 19th, 20th, 21stand / or 22ndnucleotides from 5′-end of the antisense strand.

35. The dsRNAi agent of any one of claims 18, 19, 33 and 34, wherein at least one of the 3′-PS groups in each sense strand and antisense strand has a stereopure Rp configuration.

36. The dsRNAi agent of any one of claims 18, 19, 33 and 34, wherein at least one of the 3′-PS groups in each sense strand and antisense strand has a stereopure Sp configuration.

37. A double stranded RNAi (dsRNAi) agent comprising: a sense strand having a nucleotide sequence selected from SEQ ID NOs: 812 to 1052 in Table 2 and SEQ ID NOs: 1294 to 1297, 1448 to 1462, and 1481 to 1482 in Table 3; and an antisense strand forming a duplex with the sense strand and having a nucleotide sequence selected from SEQ ID NOs: 1053 to 1293 in Table 2 and 1298 to 1301, 1463 to 1477, and 2600 to 2605 in Table 3.

38. The dsRNAi agent of any one of claims 1 through 37, further comprising a ligand. 516PAT059649-WO-PCT 39. The dsRNAi agent of claim 38, wherein the ligand comprises a N-acetylgalactosamine (GalNAc) moiety.

40. The dsRNAi agent of claim 38 or 39, wherein the ligand has a structure of: ,each L1is independently a linker which may be same or different in each occurrence; L2is a linker; n is an integer from 1 to 3; and is an attachment point to the sense strand or an antisense strand.

41. The dsRNAi agent of claim 40, wherein the ligand comprises the following structure of , or517PAT059649-WO-PCT ,each p1, p2, p3, q1, q2, r1, r2 and r3 is independently an integer from 0 to 12; each n1, n2, and n3 is independently an integer from 1 to 3; and “*” is an attachment point to L2.

42. The dsRNAi agent of claim 38 or 39, wherein the ligand has a structure of: ),each L11, L12, L13, L14, and L15is an independently a linker; L2is a linker; is an attachment point to the sense strand or the antisense strand.

43. The dsRNAi agent of claim 42, wherein the ligand has a structure of: 518PAT059649-WO-PCT 1), 2),wherein: each p11 and q11 is independently an integer from 0 to 12; each z1, z2, and z3 is independently an integer of 0 to 12; and is an attachment point to the sense strand or the antisense strand.

44. The dsRNAi agent of any one of claims 38 through 43, wherein the ligand comprises the following structure: 519PAT059649-WO-PCT orw ere n is an attachment point to the sense strand or the antisense strand. 520PAT059649-WO-PCT 45. The dsRNAi agent of claim 44, wherein the ligand is conjugated to 3′ end of the sense strand to form the following structure:wherein W is -OH or -SH.

46. The dsRNAi agent of claim 44, wherein the ligand is conjugated to 5′ end of the sense strand to form the following structure: 521PAT059649-WO-PCT ,wherein W is -OH or -SH.

47. The dsRNAi agent of claim 45 or 46, wherein W is -OH.

48. The dsRNAi agent of any one of claims 1 through 47, wherein the dsRNAi agent is in a pharmaceutically acceptable salt form.

49. The dsRNAi agent of claim 45, wherein the pharmaceutically acceptable salt is a sodium salt.

50. A pharmaceutical composition comprising the dsRNAi agent of any one of claims 1 through 49, and a pharmaceutically acceptable carrier.

51. The pharmaceutical composition of claim 50, wherein the composition is in an aqueous solution form. 522PAT059649-WO-PCT 52. The pharmaceutical composition of any of claims 50 through 51, further comprising an additional therapeutic agent selected from a proprotein convertase subtilisin kexin 9 (PCSK9) inhibitor, a lysophosphatidic acid (LPA) receptor inhibitor, an angiotensinogen (AGT) inhibitor, a fibrate, a bile acid sequestrant, niacin, an antiplatelet agent, an angiotensin converting enzyme inhibitor, an angiotensin II receptor antagonist, an acylCoA cholesterol acetyltransferase (ACAT) inhibitor, a cholesterol absorption inhibitor, a cholesterol ester transfer protein (CETP) inhibitor, a microsomal triglyceride transfer protein (MTTP) inhibitor, a cholesterol modulator, a bile acid modulator, a peroxisome proliferation activated receptor (PPAR) agonist, a gene-based therapy, a composite vascular protectant, a glycoprotein IIb / IIIa inhibitor, aspirin or an aspirin-like compound, an IBAT inhibitor, a squalene synthase inhibitor, a monocyte chemoattractant protein (MCP)-I inhibitor, and a combination thereof.

53. The pharmaceutical composition of claim 52, wherein the additional therapeutic agent comprises a PCSK9 inhibitor.

54. The pharmaceutical composition of claim 53, wherein the PCSK9 inhibitor is a second dsRNAi agent.

55. The pharmaceutical composition of claim 54, wherein the second dsRNAi agent comprises inclisiran.

56. A combination of (i) the dsRNAi agent of any one of claims 1 through 49, and (ii) a second agent selected from a proprotein convertase subtilisin kexin 9 (PCSK9) inhibitor, a lysophosphatidic acid (LPA) receptor inhibitor, an angiotensinogen (AGT) inhibitor, a fibrate, a bile acid sequestrant, niacin, an antiplatelet agent, an angiotensin converting enzyme inhibitor, an angiotensin II receptor antagonist, an acylCoA cholesterol acetyltransferase (ACAT) inhibitor, a cholesterol absorption inhibitor, a cholesterol ester transfer protein (CETP) inhibitor, a microsomal triglyceride transfer protein (MTTP) inhibitor, a cholesterol modulator, a bile acid modulator, a peroxisome proliferation activated receptor (PPAR) agonist, a gene-based therapy, a composite vascular protectant, a glycoprotein IIb / IIIa inhibitor, aspirin or an aspirin-like compound, an IBAT 523PAT059649-WO-PCT inhibitor, a squalene synthase inhibitor, a monocyte chemoattractant protein (MCP)-I inhibitor, and a combination thereof.

57. The combination of claim 56, wherein the second agent is a second dsRNAi agent.

58. The combination of claim 57, wherein the second dsRNAi agent is a dsRNA agent that targets one or more of the genes selected from the group consisting of PCSK9, LPA, AGT, ACE, ACE2, AGTR1, AGTR2, ACAT, CETP, MTTP, PPAR, IBAT, FDFT1, ERG9, SQS1, Ccl2, CCR2, CCL7, CCL8. CCL13, and CCL16.

59. The combination of any one of claims 56 to 58, wherein the second dsRNAi agent comprises inclisiran.

60. A pharmaceutical composition comprising the combination of any one of claims 56 through 59.

61. The pharmaceutical composition of claim 60, wherein the second dsRNAi agent is in a pharmaceutically acceptable salt form.

62. The pharmaceutical composition of claim 61, wherein the pharmaceutically acceptable salt of the second dsRNAi agent is a sodium salt.

63. The pharmaceutical composition of any one of claims 60 to 62, wherein the dsRNAi agent and the second agent are formulated in the same composition.

64. The pharmaceutical composition of any one of claims 60 to 63, wherein the dsRNAi agent and the second agent are formulated in the separate compositions.

65. A method of inhibiting expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) in a subject comprising: 524PAT059649-WO-PCT administering to the subject the dsRNAi agent of any one of claims 1 through 49 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 50 through 55.

66. A method of lowering a level of low-density lipoprotein cholesterol (LDL-C) in a subject, comprising: administering to the subject the dsRNAi agent of any one of claims 1 through 49 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 50 through 55.

67. A method of treating or preventing an HMGCR-associated disorder or disease in a subject, comprising: administering to the subject the dsRNAi agent of any one of claims 1 through 49 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 50 through 55.

68. The method of claim 67, wherein the HMGCR-associated disorder or disease is hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, primary hyperlipidemia, heterozygous familiar hypercholesterolemia (HeFH), homozygous familiar hypercholesterolemia (HoFH), congestive heart disease (CHD) or atherosclerosis.

69. A method of treating or preventing hyperlipidemia in a subject, comprising: administering to the subject the dsRNAi agent of any one of claims 1 through 49 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 50 through 55.

70. The method of claim 69, wherein the hyperlipidemia is hypercholesterolemia, or hypertriglyceridemia. 71 A method of treating or preventing atherosclerotic cardiovascular disease (ASCVD) in a subject, comprising: 525PAT059649-WO-PCT administering to the subject the dsRNAi agent of any one of claims 1 through 49 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 50 through 55.

72. The method of any one of claims 65 through 71, wherein the dsRNAi agent or the pharmaceutical composition is administered subcutaneously or intravenously.

73. The method of any one of claims 62 through 72, further comprising administering to the subject an additional therapeutic agent selected from a proprotein convertase subtilisin kexin 9 (PCSK9) inhibitor, a lysophosphatidic acid (LPA) receptor inhibitor, an angiotensinogen (AGT) inhibitor, a fibrate, a bile acid sequestrant, niacin, an antiplatelet agent, an angiotensin converting enzyme inhibitor, an angiotensin II receptor antagonist, an acylCoA cholesterol acetyltransferase (ACAT) inhibitor, a cholesterol absorption inhibitor, a cholesterol ester transfer protein (CETP) inhibitor, a microsomal triglyceride transfer protein (MTTP) inhibitor, a cholesterol modulator, a bile acid modulator, a peroxisome proliferation activated receptor (PPAR) agonist, a gene-based therapy, a composite vascular protectant, a glycoprotein IIb / IIIa inhibitor, aspirin or an aspirin- like compound, an IBAT inhibitor, a squalene synthase inhibitor, a monocyte chemoattractant protein (MCP)-I inhibitor, and a combination thereof.

74. The method of claim 73, wherein the additional therapeutic agent is a second dsRNAi agent.

75. The method of claim 74, wherein the second dsRNAi agent comprises a PCSK9 inhibitor.

76. The method of claim 75, wherein the second dsRNAi agent comprises inclisiran.

77. The method of any one of claims 73 through 76, wherein the dsRNAi agent or the pharmaceutical composition and the additional therapeutic agent are administered simultaneously. 526PAT059649-WO-PCT 78. The method of any one of claims 73 through 76, wherein the dsRNAi agent or the pharmaceutical composition and the additional therapeutic agent are administered subsequently.

79. The method of claim 78, wherein the dsRNAi agent is administered before administering the additional therapeutic agent.

80. The method of claim 78, wherein the additional therapeutic agent is administered before administering the dsRNAi agent.

81. The method of any one of claims 73 through 80, wherein the additional therapeutic agent is administered subcutaneously or intravenously.

82. The method of any one of claims 65 through 81, wherein the subject is a human.

83. The method of any one of claims 65 through 82, wherein the subject has or is diagnosed with hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, primary hyperlipidemia, heterozygous familiar hypercholesterolemia (HeFH), homozygous familiar hypercholesterolemia (HoFH), congestive heart disease (CHD) or atherosclerosis.

84. The method of any one of claims 65 through 83, wherein the subject does not have a muscle side effect after the administrating the dsRNAi agent of any one of claims 1 through 49 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 50 through 55.

85. A method of lowering a level of low-density lipoprotein cholesterol (LDL-C) in a subject, comprising: administering to the subject the pharmaceutical composition of any one of claims 60 through 64.

86. A method of treating or preventing an HMGCR-associated disorder or disease in a subject, comprising: 527PAT059649-WO-PCT administering to the subject the pharmaceutical composition of any one of claims 60 through 64.

87. The method of claim 86, wherein the HMGCR-associated disorder or disease is hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, primary hyperlipidemia, heterozygous familiar hypercholesterolemia (HeFH), homozygous familiar hypercholesterolemia (HoFH), congestive heart disease (CHD) or atherosclerosis.

88. A method of treating or preventing hyperlipidemia in a subject, comprising: administering to the subject the pharmaceutical composition of any one of claims 60 through 64.

89. A method of treating or preventing atherosclerotic cardiovascular disease (ASCVD) in a subject, comprising: administering to the subject the pharmaceutical composition of any one of claims 60 through 64.

90. The method of any one of claims 85 through 89, wherein the dsRNAi agent and the second agent is administered subcutaneously or intravenously.

91. The method of any one of claims 85 through 89, wherein the dsRNAi agent and the second agent are administered simultaneously.

92. The method of any one of claims 85 through 91, wherein the dsRNAi agent and the second agent are administered subsequently.

93. The method of claim 92, wherein the dsRNAi agent is administered before administering the second agent.

94. The method of claim 92, wherein the second agent is administered before administering the dsRNAi agent. 528PAT059649-WO-PCT 95. The method of any one of claims 85 through 94, wherein the subject is a human.

96. The method of any one of claims 85 through 95, wherein the subject has or is diagnosed with hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, primary hyperlipidemia, heterozygous familiar hypercholesterolemia (HeFH), homozygous familiar hypercholesterolemia (HoFH), congestive heart disease (CHD) or atherosclerosis.

97. The method of any one of claims 85 through 96, wherein the subject does not have a muscle side effect after the administrating the pharmaceutical composition of any one of claims 60 through 64.

98. A method of reducing the risk of a major adverse cardiovascular event in a subject, comprising administering to the subject the dsRNAi agent of any one of claims 1 through 49 or a pharmaceutically acceptable salt thereof, the pharmaceutical composition of any one of claims 50 through 55, the combination of any one of claims 56 through 59, or the pharmaceutical composition of any one of claims 60 through 64.

99. The method of claim 98, wherein the major adverse cardiovascular event is cardiovascular death, non-fatal myocardial infarction, non-fatal ischemic stroke, or urgent coronary revascularization.

100. The method of claim 98, wherein the subject has an established cardiovascular disease.

101. The method of claim 98, where the subject has not experienced a major atherosclerotic cardiovascular disease (ASCVD) event.

102. A kit comprising the dsRNAi agent of any one of claims 1 through 49 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 50 through 55. 529PAT059649-WO-PCT 103. The kit of claim 102, further comprising an additional therapeutic agent selected from a proprotein convertase subtilisin kexin 9 (PCSK9) inhibitor, a fibrate, a bile acid sequestrant, niacin, an antiplatelet agent, an angiotensin converting enzyme inhibitor, an angiotensin II receptor antagonist, an acylCoA cholesterol acetyltransferase (ACAT) inhibitor, a cholesterol absorption inhibitor, a cholesterol ester transfer protein (CETP) inhibitor, a microsomal triglyceride transfer protein (MTTP) inhibitor, a cholesterol modulator, a bile acid modulator, a peroxisome proliferation activated receptor (PPAR) agonist, a gene-based therapy, a composite vascular protectant, a glycoprotein IIb / IIIa inhibitor, aspirin or an aspirin-like compound, an IBAT inhibitor, a squalene synthase inhibitor, a monocyte chemoattractant protein (MCP)-I inhibitor, and a combination thereof.

104. The kit of claim 103, wherein the additional therapeutic agent is a second dsRNAi agent.

105. The kit of claim 104, wherein the second dsRNAi agent is a dsRNA agent that targets one or more of the genes selected from the group consisting of PCSK9, LPA, AGT, ACE, ACE2, AGTR1, AGTR2, ACAT, CETP, MTTP, PPAR, IBAT, FDFT1, ERG9, SQS1, Ccl2, CCR2, CCL7, CCL8. CCL13, and CCL16.

106. The kit of claim 105, wherein the second dsRNAi agent comprises the PCSK9 inhibitor.

107. The kit of claim 106, wherein the second dsRNAi agent comprises inclisiran.

108. The kit of any one of claims 103 through 107, wherein the dsRNAi agent and the additional therapeutic agent are contained in a single vial.

109. The kit of any one of claims 103 through 107, wherein the dsRNAi agent and the additional therapeutic agent are contained in separate vials. 530PAT059649-WO-PCT 110. The kit of any one of claims 102 through 109, further comprising one or more applicators.

111. The kit of claim 110, wherein the one or more applicators comprises a syringe.

112. A kit comprising the pharmaceutical composition of any one of claims 60 through 64.

113. The kit of claim 112, wherein the dsRNAi agent and the second agent are contained in a single vial.

114. The kit of claim 112, wherein the dsRNAi agent and the second agent are contained in separate vials.

115. The kit of any one of claims 103 through 114, further comprising one or more applicators.

116. The kit of claim 115, wherein the one or more applicators are syringes. 531

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