Double stranded rnai agents, compositions and methods of use

Abstract

Disclosed are, inter alia, double stranded RNAi (dsRNAi) agents inhibiting expression of proprotein convertase subtilisin kexin 9 (PCSK9), for example, human PCSK9, compositions including the same, and methods of treatment using the same.

Description

PAT059857-PCT-SEC01DOUBLE STRANDED RNAi AGENTS, COMPOSITIONS AND METHODS OF USECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims priority and benefit to the U. S. Patent Application No.63 / 724,671 filed November 25, 2024, the disclosure of which is incorporated herein by reference in its entirety.SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on October 10, 2025, is named PAT059857-PCT-SEC01_SL.xml and is 4,029,476 bytes in size.TECHNICAL FIELD

[0003] The present disclosure provides, inter alia, double stranded RNAi (dsRNAi) agents inhibiting expression of proprotein convertase subtilisin kexin 9 (PCSK9), for example, human PCSK9, compositions including the same, and methods of treatment using the same.BACKGROUND

[0004] 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.

[0005] Proprotein convertase subtilisin kexin 9 (PCSK9) is a member of the subtilisin serine protease family and is involved in cholesterol metabolism and homeostasis. A liver cell uptakes LDL-C via low-density lipoprotein receptor (LDLR) to directly remove such atherosclerotic lipoproteins from the plasma via uptake into a liver cell. If the level of LDLR decreases, LDL-C level increases in circulation, and the elevated circulating LDL-C level causes increased deposition in the arteries of LDL particles and the cholesterol they carry, which promotes the formation and progression of atherosclerotic plaques in the artery. PCSK9 binds to hepatic LDLR to induce endocytosis and lysosomal degradation of LDLR in the liver cell, thereby reducing LDL-C uptakePAT059857-PCT-SEC01by LDLR. PCSK9 inhibitors have been clinically proven to reduce LDLR degradation by inhibiting PCSK9 expression / function and lower LDL-C levels in the general population.

[0006] However, there is still an unmet need in the art for LDL-C lowering treatments that have increased potency and / or durability of action.SUMMARY OF THE INVENTION

[0007] Provided herein are, inter alia, compounds that can inhibit expression of PCSK9 in a subject, for example, e.g., in liver cells of a subject.

[0008] In an aspect, the disclosure provides a double stranded RNAi (dsRNAi) agent comprising:(i) a sense strand comprising a nucleotide sequence selected from SEQ ID Nos. 3 to 381; (ii) an antisense strand forming a duplex with the sense strand and comprising a nucleotide sequence selected from SEQ ID Nos. 382 to 760.

[0009] In an aspect, the disclosure provides a double stranded RNAi (dsRNAi) agent comprising:(i) a sense strand comprising a nucleotide sequence selected from SEQ ID Nos. 761 to 776;(ii) an antisense strand forming a duplex with the sense strand and comprising a nucleotide sequence selected from SEQ ID Nos. 777 to 792.

[0010] In some embodiments, one or more nucleotides in the sense strand and the antisense strand are modified nucleotides. In some embodiments, all nucleotides in the sense strand and the antisense strand are modified nucleotides.

[0011] 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 modificationPAT059857-PCT-SEC01containing 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 2-0-(N-methylacetamide) modification.

[0012] In some embodiments, each of the modified nucleotides independently comprises one or more modifications selected from 2'-deoxy modification, 2'-O-alkoxyalkyl modification, 2'-0-alkyl modification, 2'-0-allyl modification, 2'-C-allyl modification, 2'-halo modification, modification containing a non-natural nucleobase, GNA modification, and TNA modification.

[0013] In some embodiments, the dsRNAi agent comprises a 3'-phosphorothioate (PS) modification.

[0014] In some embodiments, each of the modified nucleotides independently comprises one or more modifications selected from 2'-deoxy modification, 2'-O-methyl (2'-0Me) modification, 2'-fluoro (2'-F) modification, 2'-O-methoxyethyl (2'-M0E) modification, the modification containing a non-natural nucleobase, TNA, GNA, 3'-phosphorothioate (PS) modification, and 5'-vinyl-phosphonate (5'-VP) modification.

[0015] In some embodiments, the sense strand comprises one or two 2'-MOE modifications positioned at the 1st and / or 2nd nucleotides from the 5' end.

[0016] In some embodiments, the sense strand comprises one or two 2'-MOE modifications positioned at the 1st and / or 2nd nucleotides from the 3' end.

[0017] In some embodiments, the sense strand comprises one or two TNAs positioned at the 1st and / or 2nd nucleotides from the 5' end.

[0018] In some embodiments, the sense strand comprises one or two TNAs positioned at the 1st and / or 2nd nucleotides from the 3' end.

[0019] In some embodiments, the antisense strand comprises a 5'-VP modification at the 1st nucleotide from the 5' end.

[0020] In some embodiments, the antisense strand comprises a 5'-VP-2'-OMe modification at the 1st position from the 5' end.

[0021] In some embodiments, each of the sense strand and the antisense strand independently comprises two, three, four, five or six 2'-F modified nucleotides.

[0022] In some embodiments, the sense strand comprises one or two 3'-PS modifications at the 1st and / or 2nd nucleotides from the 5' end.PAT059857-PCT-SEC01

[0023] In some embodiments, the sense strand comprises one or two 3'-PS modifications at the 1st and / or 2nd nucleotides from the 3' end.

[0024] In some embodiments, the antisense strand comprises one or two 3'-PS modifications at the 1st and / or 2nd nucleotides from the 5' end, and / or one or two 3'-PS modifications at the 1st and / or 2nd nucleotides from the 3' end.

[0025] In certain aspects, the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.

[0026] In some embodiments, the sense strand comprises one to four 2'-MOE modifications positioned at the 1st, 2nd, 20th, and / or 21st nucleotides from the 5' end.

[0027] In some embodiments, the sense strand does not comprise a 2'-MOE modification at the 3rd to 19th positions from the 5' end.

[0028] In some embodiments, the sense strand comprises one to four TNAs positioned at the 1st, 2nd, 20th, and / or 21st nucleotides from the 5' end.

[0029] In some embodiments, the sense strand does not comprise a 2'-MOE modifications and TNA at the 3rd to 19th positions from the 5' end.

[0030] In some embodiments, the sense strand comprises two, three, or four 2'-F modifications positioned at the 7th, 9th, 10th, and / or 11th nucleotides from the 5' end.

[0031] In some embodiments, the sense strand comprises one or two 2'-deoxy modifications positioned at the 10th and / or 11th nucleotides from the 5' end.

[0032] In some embodiments, the sense strand comprises (i) 2'-F modifications positioned at the 7th, 9th, and 10th nucleotides from the 5' end and (ii) a 2'-deoxy modification positioned at the 11th nucleotide from the 5' end.

[0033] In some embodiments, the remaining nucleotides in the sense strand comprise 2'-OMe modifications.

[0034] In some embodiments, the antisense strand comprises a 5'-(E)-VP modification at the 1st nucleotide from the 5' end.

[0035] In some embodiments, the antisense strand comprises a 5'-(E)-VP-2'-OMe modification at the 1st nucleotide from the 5' end.PAT059857-PCT-SEC01

[0036] In some embodiments, the antisense strand comprises two, three, or four 2'-F modifications positioned at the 2nd, 6th, 14th, and / or 16th nucleotides from the 5' end.

[0037] In some embodiments, the antisense strand comprises 2'-F modifications positioned at the 2nd, 6th, 14th, and 16th nucleotides from the 5' end.

[0038] In some embodiments, the antisense strand comprises 2'-F modifications positioned at the 2nd, 6th, 14th, and 16th nucleotides from the 5' end and a TNA positioned at the 3rd nucleotide from the 5' end.

[0039] In some embodiments, the antisense strand comprises 2'-F modifications positioned at the 2nd, 6th, 14th, and 16th nucleotides from the 5' end and a TNA, GNA or 2'-deoxy modification positioned at the 5th nucleotide from the 5' end.

[0040] In some embodiments, the antisense strand comprises 2'-F modifications positioned at the 2nd, 14th, and 16th nucleotides from the 5' end and a TNA, GNA or 2'-deoxy modification positioned at the 6th nucleotide from the 5' end.

[0041] In some embodiments, the antisense strand comprises 2'-F modifications positioned at the 2nd, 6th, 14th, and 16th nucleotides and a TNA, GNA or 2'-deoxy modification positioned at the 7th nucleotide.

[0042] In some embodiments, the remaining nucleotides in antisense strand comprise 2'-OMe modified modifications.

[0043] In some embodiments, the sense strand comprises one to eight 3'-PS group at the 1st, 2nd, 3rd, 4th, 17th, 18th, 19th and / or 20th nucleotides from the 5' end.

[0044] In some embodiments, the antisense strand comprises one to eight 3'-PS group at the 1st, 2nd, 3rd, 4th, 19th, 20th, 21st and / or 22nd nucleotides from the 5' end.

[0045] In some embodiments, at least one of the 3'-PS groups in each sense strand and antisense strand has a stereopure Rp configuration.

[0046] In some embodiments, at least one of the 3'-PS groups in each sense strand and antisense strand has a stereopure Sp configuration.

[0047] In certain aspects, the double stranded RNAi (dsRNAi) agent comprises:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andPAT059857-PCT-SEC01an antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andan antisense strand comprising SEQ ID NO: 859;(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;or(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883.

[0048] In some embodiments, the dsRNAi agent comprises a ligand.

[0049] In some embodiments, the ligand comprises a N-acetylgalactosamine (GalNAc) moiety.

[0050] In some embodiments, the ligand has a structure of:wherein: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; andis an attachment point to the sense strand or the antisense strand, or to a conjugate linker conjugated to the sense strand or the antisense strand.

[0051] In some embodiments, the ligand comprises the following structure ofPAT059857-PCT-SEC01wherein:each pl, p2, p3, ql, q2, rl, r2 and r3 is independently an integer from 0 to 12;each nl, n2, and n3 is independently an integer from 1 to 3; andis an attachment point to L2.

[0052] In some embodiments, the ligand has a structure of:NHAcPAT059857-PCT-SEC01wherein: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, or to a conjugate linker conjugated to the sense strand or the antisense strand.

[0053] In some embodiments, the ligand has a structure of:HO OHHO OHwherein:each pl 1 and ql 1 is independently an integer from 0 to 12;each zl, z2, and z3 is independently an integer of 0 to 12; andPAT059857-PCT-SEC01is an attachment point to the sense strand or the antisense strand, or to a conjugate linker conjugated to the sense strand or the antisense strand.

[0054] In some embodiments, the ligand comprises the following structure:orPAT059857-PCT-SEC01whereinis an atachment point to the sense strand or the antisense strand or to a conjugate linker conjugated to the sense strand or the antisense strand.

[0055] In some embodiments, the ligand is conjugated to 3' end of the sense strand to form the following structure:PAT059857-PCT-SEC01OHwherein W is -OH or -SH.

[0056] In some embodiments, the ligand is conjugated to 5' end of the sense strand to form the following structure:PAT059857-PCT-SEC01or a pharmaceutically acceptable salt thereof, wherein W is -OH or -SH.

[0057] In some embodiments, W is -OH.

[0058] In an aspect, a double stranded RNAi (dsRNAi) agent comprises:(a) a sense strand consisting of SEQ ID NO: 906, andan antisense strand consisting of SEQ ID NO: 963;(b) a sense strand consisting of SEQ ID NO: 907, andan antisense strand consisting of SEQ ID NO: 964;(c) a sense strand consisting of SEQ ID NO: 947, andan antisense strand consisting of SEQ ID NO: 1004;(d) a sense strand consisting of SEQ ID NO: 948, andan antisense strand consisting of SEQ ID NO: 1005;(e) a sense strand consisting of SEQ ID NO: 949, andan antisense strand consisting of SEQ ID NO: 1006;or(f) a sense strand consisting of SEQ ID NO: 950, andan antisense strand consisting of SEQ ID NO: 1007;wherein the ligand (L96) is conjugated to the 3' end of the sense strand to form the following schematic:PAT059857-PCT-SEC01OHor a pharmaceutically acceptable salt thereof, wherein W is -OH.

[0059] In some embodiments, the dsRNAi agent is in a pharmaceutically acceptable salt form.

[0060] In some embodiments, the pharmaceutically acceptable salt is a sodium salt.

[0061] In an aspect, the disclosure provides a pharmaceutical composition comprising the dsRNAi agent as described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0062] In some embodiments, the composition is in an aqueous solution form.

[0063] In an aspect, the disclosure provides a method of inhibiting PCSK9 expression in a cell, the method comprising:(a) contacting the cell with the dsRNAi agent as described herein or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described herein; and (b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a PCSK9 gene, thereby inhibiting expression of the PCSK9 gene in the cell.

[0064] In an aspect, the disclosure provides a method of lowering a level of low-density lipoprotein cholesterol (LDL-C) in a subject in need thereof, comprising administering to thePAT059857-PCT-SEC01subject the dsRNAi agent as described herein or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described herein.

[0065] In an aspect, the disclosure provides a method of treating lipidemia mediated by PCSK9 expression in a subject in need thereof, comprising administering to the subject the dsRNAi agent as described herein or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described herein.

[0066] In an aspect, the disclosure provides a method of treating or preventing atherosclerotic cardiovascular disease (ASCVD) in a subject in need thereof, comprising administering to the subject the dsRNAi agent as described herein, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described herein.

[0067] In an aspect, the disclosure provides a method of reducing or preventing cardiovascular event in a subject in need thereof, comprising administering to the subject the dsRNAi agent as described herein or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described herein.

[0068] In some embodiments, the cardiovascular event is cardiovascular death, non-fatal myocardial infarction (MI), non-fatal ischemic stroke, urgent coronary revascularization, coronary heart disease (CHD) death, or any combination thereof.

[0069] In an aspect, the disclosure provides a method of reducing or preventing a major limb adverse event (MALE) in a subject in need thereof, comprising administering to the subject the dsRNAi agent as described herein or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described herein.

[0070] In some embodiments, the MALE is acute lower limb ischemia, lower limb amputation due to ischemia, urgent lower limb revascularization for ischemia, or any combination thereof.

[0071] In an aspect, the disclosure provides a method of inhibiting PCSK9 expression in a cell, the method comprising:(i) contacting the cell with a dsRNAi agent, wherein the dsRNAi agent comprises:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andPAT059857-PCT-SEC01an antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andan antisense strand comprising SEQ ID NO: 859;(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;or(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883,(ii) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a PCSK9 gene, thereby inhibiting expression of the PCSK9 gene in the cell.

[0072] In an aspect, the disclosure provides a method of lowering a level of low-density lipoprotein cholesterol (LDL-C) in a subject in need thereof, comprising administering to the subject a dsRNAi agent, wherein the dsRNAi agent comprises:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andan antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andan antisense strand comprising SEQ ID NO: 859;(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;or(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883.PAT059857-PCT-SEC01

[0073] In an aspect, the disclosure provides a method of treating lipidemia mediated by PCSK9 expression in a subject in need thereof, comprising administering to the subject a dsRNAi agent, wherein the dsRNAi agent comprises:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andan antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andan antisense strand comprising SEQ ID NO: 859;(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;or(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883.

[0074] In an aspect, the disclosure provides a method of treating or preventing atherosclerotic cardiovascular disease (ASCVD) in a subject in need thereof, comprising administering to the subject a dsRNAi agent, wherein the dsRNAi agent comprises:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andan antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andan antisense strand comprising SEQ ID NO: 859;(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;orPAT059857-PCT-SEC01(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883.

[0075] In an aspect, the disclosure provides a method of reducing or preventing cardiovascular event in a subject in need thereof, comprising administering to the subject a dsRNAi agent, wherein the dsRNAi agent comprises:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andan antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andan antisense strand comprising SEQ ID NO: 859;(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;or(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883.

[0076] In some embodiments, the cardiovascular event is cardiovascular death, non-fatal myocardial infarction (MI), non-fatal ischemic stroke, urgent coronary revascularization, coronary heart disease (CHD) death, or any combination thereof.

[0077] In an aspect, the disclosure provides a method of reducing or preventing a major limb adverse event (MALE) in a subject in need thereof, comprising administering to the subject a dsRNAi agent, wherein the dsRNAi agent comprises:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andan antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andan antisense strand comprising SEQ ID NO: 859;PAT059857-PCT-SEC01(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;or(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883.

[0078] In some embodiments, the MALE is acute lower limb ischemia, lower limb amputation due to ischemia, urgent lower limb revascularization for ischemia, or any combination thereof.

[0079] In some embodiments, the dsRNAi agent further comprises a ligand comprising the following structure:orPAT059857-PCT-SEC01whereinis an atachment point to the sense strand or the antisense strand or to a conjugate linker conjugated to the sense strand or the antisense strand.

[0080] In some embodiments, the subject is a human.

[0081] In some embodiments, the subject has or is diagnosed with hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, congestive heart disease (CHD) or atherosclerosis.

[0082] In some embodiments, the dsRNAi agent or the pharmaceutical composition is administered to the subject subcutaneously or intravenously.

[0083] In an aspect, the disclosure provides a kit comprising the dsRNAi agent as described herein, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described herein.

[0084] In some embodiments, the kit further comprises an applicator.

[0085] In some embodiments, the applicator is a syringe.

[0086] In some embodiments, the applicator is a pre-filled syringe.

[0087] Other aspects of the invention are disclosed infra.PAT059857-PCT-SEC01BRIEF DESCRIPTION OF DRAWINGS

[0088] Figures 1A-1B. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs DI, D2, D3, D4, D5, or D6 to humanized PCSK9 mice on (Figure 1A) plasma PCSK9 levels over time and (Figure 1A) liver human PCSK9 mRNA level, liver siRNA level, and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 56 post-dose.

[0089] Figures 2A-2B. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs D7, D8, D9, or DI 0 to humanized PCSK9 mice on (Figure 3 A) plasma PCSK9 levels over time and (Figure 3B) liver human PCSK9 mRNA level, liver siRNA level, and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 77post-dose.

[0090] Figures 3A-3B. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs DI, D4, Dll, or D12 to humanized PCSK9 mice on (Figure 3A) plasma PCSK9 levels over time and (Figure 3B) liver human PCSK9 mRNA level, liver siRNA level, and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 63 post-dose.

[0091] Figures 4A-4B. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs D4, D14, DI 5, D16, or D17 to humanized PCSK9 mice on (Figure 4A) plasma PCSK9 levels over time and (Figure 4B) liver human PCSK9 mRNA level, liver siRNA level, and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 77 post-dose.

[0092] Figures 5A-5B. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs D4, D18, D19, D20, D21, D22, or D23 to humanized PCSK9 mice on (Figure 5A) plasma PCSK9 levels over time and (Figure 5B) liver human PCSK9 mRNA level, liver siRNA level, and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 77 post-dose.

[0093] Figures 6A-6B. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs D4, DI 3, D24, D25, D26, D27, or D28 to humanized PCSK9 mice on (Figure 6A) plasma PCSK9 levels over time and (Figure 6B) liver human PCSK9 mRNA level, liver siRNA level, and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 63 post-dose.

[0094] Figures 7A-7B. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs D25, D29, D30, D31, D32, D33, or D34 to humanized PC SK9 mice on (Figure 7A) plasma PCSK9 levels over time and (Figure 7B) liver human PCSK9 mRNA level, liver siRNA level, and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 64 post-dose.PAT059857-PCT-SEC01

[0095] Figures 8A-8B. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs D4, D15, D35, D36, D37, or D38 to humanized PCSK9 mice on (Figure 8A) plasma PCSK9 levels over time and (Figure 8B) liver human PCSK9 mRNA level, liver siRNA level, and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 77 post-dose.

[0096] Figure 9 shows example PCSK9 siRNA compounds (dsRNAi agents) as described herein. Figure discloses SEQ ID NOS 963, 906, 964, 907, 1004, 947, 1005, 948, 1006, 949, 1007, and 950, respectively, in order of appearance (from top to bottom).

[0097] Figures 10A-10C. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs D15, D38, D39, D40, D41, D42, D43, D44, D45, D46, D47, or D48 to humanized PCSK9 mice on plasma PCSK9 levels over time (Figures 10A-10B) and liver siRNA level and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 77 post-dose (Figure 10C).

[0098] Figures 11A-11B. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs D4, DI 9, D49, or D50 to humanized PCSK9 mice on plasma PCSK9 levels over time (Figure 11 A) and liver human PCSK9 mRNA level, liver siRNA level, and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 77 post-dose (Figure 1 IB).

[0099] Figures 12A-12C. Effects of a single subcutaneous administration of PCSK9 siRNAs D7 (3 or 6 mg / kg), D8 (3 or 6 mg / kg), D51 (3 mg / kg), D52 (3 mg / kg), D53 (3 mg / kg), or D54 (1, 3, or 6 mg / kg) to humanized PCSK9 mice on plasma PCSK9 levels over time (Figures 12A-12B) and liver human PCSK9 mRNA level, liver siRNA level, and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 77 post-dose (Figure 12C).

[0100] Figures 13A-13C. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs D38, D55, D42, D43, D44, D56, D57, D58, or D59 to humanized PCSK9 mice on plasma PCSK9 levels over time (Figures 13A-13B) and liver human PCSK9 mRNA level, liver siRNA level, and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 77 postdose (Figure 13C).

[0101] Figures 14A-14B. Effects of a single 0.3, 1, or 3 mg / kg subcutaneous administration of PCSK9 siRNAs D7 or D52 to humanized PCSK9 mice on plasma PCSK9 levels over time (Figure 14A) and liver human PCSK9 mRNA level, liver siRNA level, and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 77 post-dose (Figure 14B).PAT059857-PCT-SEC01

[0102] Figures 15A-15B. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs D52, D60, D61, or D62 to humanized PCSK9 mice on plasma PCSK9 levels over time (Figure 15A) and liver siRNA level and liver RISC loading (i.e., the level of siRNA incorporated into RISC) on day 77 post-dose (Figure 15B).

[0103] Figure 16. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs D8 or D63 to humanized PCSK9 mice on plasma PCSK9 levels over time.

[0104] Figure 17. Effects of PCSK9 siRNAs D7, D8, or D52 transfection at various concentrations into cultured Hep3B cells on PCSK9 mRNA levels.

[0105] Figure 18. Results of in vitro specificity profiling of the PCSK9 siRNAs D52, D60, D61, or D62 in Hep3B cells.

[0106] Figures 19A-19B. Effects of a single 3 mg / kg subcutaneous administration of PCSK9 siRNAs D8, D51, D52, D53, or D54 to obese cynomolgus monkeys on plasma PCSK9 levels over time (Figure 19A) and serum low density lipoprotein cholesterol (LDL-C) levels over time (Figure 19B).DETAILED DESCRIPTIONDefinitions

[0107] 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.

[0108] 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.

[0109] 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 thePAT059857-PCT-SEC01phrases “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 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.

[0110] 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.”

[0111] 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.

[0112] The term “nucleotide” means a compound including a nucleoside and a phosphate group (or 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., Xi to X21 and Xv to X23’, a nucleotide is meantPAT059857-PCT-SEC01by 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).

[0113] The term “nucleoside” means a monomer consisting of a nucleobase and a pentofuranosyl sugar (e.g., ribose or deoxyribose). A nucleoside including a ribose sugar ring has HO Baseto a structure of OH OH or a pharmaceutically acceptable salt thereof, and a nucleotide HO^ Baseincluding a deoxyribose sugar ring has a structure of OH or a pharmaceutically acceptable salt, wherein in each structure, “Base” is a nucleobase.

[0114] 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), deazaadenine 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 isPAT059857-PCT-SEC01adenine guanine thymine uracilor methylateduracilorcytidine 4-methyl 5-methylcytidine cytidinewhich may be optionally substituted or modified, wherein “ ‘Tz- ” denotes the point of attachment to a pentofuranosyl sugar ring (e.g., 1' position).

[0115] The term “phosphate,” or “phosphate group” as used herein a chemical species made oII\ > o"of one phosphorus atom and four oxygen atomsO, or esters, salts, or acids thereof. In certain aspects, when the phosphate groups are positioned 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,” “phosphodi ester linkage,” or “linkage.” For example, the phosphate or phosphodi ester linkage in s*° 7 othe backbone of RNA or DNA may have the structuresof or esters, salts (e.g.,PAT059857-PCT-SEC01C)\=oopharmaceutically acceptable salts), or acids (e.g., -r ) thereof, wherein “ ^’’ denotes the point of attachment to pentofuranosyl sugar rings (e.g., 5' and 3' positions) in adjacent nucleosides. In certain aspects, a variant of a phosphate 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 vinyl phosphonate (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.

[0116] 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 ofor a pharmaceutically acceptable salt thereof. Likewise, an unmodified DNA (or “deoxyribonucleotides”) in a chain of nucleotides (e.g., genomic DNA or cDNA) as disclosed herein specifically refers to a structure ofPAT059857-PCT-SEC01O=P - OHor a pharmaceutically acceptable salt thereof. In each structure “Base” is a nucleobase and > is an attachment point to the adjacent nucleotides.

[0117] 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 HC BaseO OH IO=P — OHnucleotide has a structure of,Or a pharmaceutically acceptable salt thereof. Likewise, when an unmodified DNA is the first nucleotide from the 5' end of a DNA chain (e.g.,o=P — OH genomic DNA or cDNA), that nucleotide has a structure of, or a pharmaceutically acceptable salt thereof. In each structure “Base” is a nucleobase andis an attachment point (5' oxygen) to the adjacent nucleotides. Alternatively but equivalently, for example, the first nucleotide from the 5' end of an RNA chain (e.g., mRNA, or sense strand orantisense strand of siRNA), that nucleotide has a structure of or a pharmaceutically acceptable salt thereof and the first nucleotide from the 5' end of a DNA chainPAT059857-PCT-SEC010=P — OH (e.g., genomic DNA or cDNA), that nucleotide has a structure of, or a pharmaceutically acceptable salt thereof, whenis an attachment point (5' oxygen) to the adjacent nucleotides.

[0118] 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), thatnucleotide has a structure of OH, or a pharmaceutically 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 has a structure of or a pharmaceutically acceptable salt thereof. In certain embodiments, 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 does not include 3' end phosphate group or phosphodi ester linkage, for example, which has been removed during hydrolysis or synthesis, has a structure ofBaseOH OH, or a pharmaceutically 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 duringPAT059857-PCT-SEC01Basehydrolysis or synthesis, has a structure of OH, or a pharmaceutically acceptable salt thereof. In each structure “Base” is a nucleobase andis an attachment point (e.g., phosphorus of the phosphate linkage) to the adjacent nucleotides.

[0119] 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.

[0120] 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), singlestranded 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.

[0121] 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-induced silencing complex (RISC) pathway. An RNAi agent directs the sequence-specific degradation of mRNAPAT059857-PCT-SEC01through 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).

[0122] 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 aspects 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.

[0123] 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.

[0124] The term “sense strand,” as used herein, refers an oligonucleotide that is complementary (e.g., partially, substantially, or fully complementary) to the antisense strand. The sense strand is typically degraded following incorporation of the antisense strand into RISC. ThePAT059857-PCT-SEC01sense 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.

[0125] 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%.

[0126] 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., double stranded RNA) are substantially or fully complementary over their entire lengths. In some embodiments, a sense strand and anPAT059857-PCT-SEC01antisense 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 singlestranded nucleotides.

[0127] 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).

[0128] 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.PAT059857-PCT-SEC01

[0129] 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 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.

[0130] 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.

[0131] 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, andPAT059857-PCT-SEC01(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 phosphodi ester) linkage or replacing a phosphate (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 of, bicyclic or bridged 7o -o ring on the ribose such as locked nucleic acid (LNA) having a structure of 7, or the like. In certain aspects, the modification is present in a nucleobase (e.g., A, G, C, T, or U) by chemical modification in a nucleobase by replacing the nucleobase with other 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.PAT059857-PCT-SEC01

[0132] 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.

[0133] 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.

[0134] Throughout the disclosure, nucleotide positions or coordinates are relative to the beginning (5' end) of the reference transcript.

[0135] 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.

[0136] ‘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 aPAT059857-PCT-SEC01dsRNA that is double-stranded over its entire length, i.e., no nucleotide overhang at either end of the molecule.

[0137] 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).

[0138] The term “non-end” herein refers to a position between the 3' end and the 5' end of the sense or antisense strand.

[0139] The term “PCSK9,” refers to a proprotein convertase subtilisin kexin 9 gene or a protein encoded by that gene. PCSK9 is also known as FH3, PC9, FHCL3, NARC1, LDLCQ1, NARC-1, or HCHOLA3. The PCSK9 gene as used herein includes human PCSK9 (e.g., Gene ID: 255738; GenBank Accession No. NM_174936.3 or NM_174936.4), mouse PCSK9 gene (e.g., Gene ID: 100102; GenBank Accession No. NM_153565.2), and dog PCSK9 gene (e.g., Gene ID: 102152231; GenBank Accession No. XM_038667514.1). Additional examples ofPCSK9 mRNA sequences from different species and variants are readily available using, e.g., GenBank.

[0140] Homo sapiens PCSK9, transcript variant 1, mRNA: GenBank: NM 174936.3 (SEQ ID NO: 1)1 gtccgatggg gctctggtgg cgtgatctgc gcgccccagg cgtcaagcac ccacacccta61 gaaggtttcc gcagcgacgt cgaggcgctc atggttgcag gcgggcgccg ccgttcagtt121 cagggtctga gcctggagga gtgagccagg cagtgagact ggctcgggcg ggccgggacg 181 cgtcgttgca gcagcggctc ccagctccca gccaggattc cgcgcgcccc ttcacgcgcc241 ctgctcctga acttcagctc ctgcacagtc ctccccaccg caaggctcaa ggcgccgccg301 gcgtggaccg cgcacggcct ctaggtctcc tcgccaggac agcaacctct cccctggccc361 tcatgggcac cgtcagctcc aggcggtcct ggtggccgct gccactgctg ctgctgctgc421 tgctgctcct gggtcccgcg ggcgcccgtg cgcaggagga cgaggacggc gactacgagg 481 agctggtgct agccttgcgt tccgaggagg acggcctggc cgaagcaccc gagcacggaa 541 ccacagccac cttccaccgc tgcgccaagg atccgtggag gttgcctggc acctacgtgg601 tggtgctgaa ggaggagacc cacctctcgc agtcagagcg cactgcccgc cgcctgcagg 661 cccaggctgc ccgccgggga tacctcacca agatcctgca tgtcttccat ggccttcttcPAT059857-PCT-SEC01721 ctggcttcct ggtgaagatg agtggcgacc tgctggagct ggccttgaag ttgccccatg781 tcgactacat cgaggaggac tcctctgtct ttgcccagag catcccgtgg aacctggagc841 ggattacccc tccacggtac cgggcggatg aataccagcc ccccgacgga ggcagcctgg 901 tggaggtgta tctcctagac accagcatac agagtgacca ccgggaaatc gagggcaggg 961 tcatggtcac cgacttcgag aatgtgcccg aggaggacgg gacccgcttc cacagacagg 1021 ccagcaagtg tgacagtcat ggcacccacc tggcaggggt ggtcagcggc cgggatgccg 1081 gcgtggccaa gggtgccagc atgcgcagcc tgcgcgtgct caactgccaa gggaagggca 1141 cggttagcgg caccctcata ggcctggagt ttattcggaa aagccagctg gtccagcctg1201 tggggccact ggtggtgctg ctgcccctgg cgggtgggta cagccgcgtc ctcaacgccg 1261 cctgccagcg cctggcgagg gctggggtcg tgctggtcac cgctgccggc aacttccggg 1321 acgatgcctg cctctactcc ccagcctcag ctcccgaggt catcacagtt ggggccacca 1381 atgcccaaga ccagccggtg accctgggga ctttggggac caactttggc cgctgtgtgg 1441 acctctttgc cccaggggag gacatcattg gtgcctccag cgactgcagc acctgctttg1501 tgtcacagag tgggacatca caggctgctg cccacgtggc tggcattgca gccatgatgc 1561 tgtctgccga gccggagctc accctggccg agttgaggca gagactgatc cacttctctg1621 ccaaagatgt catcaatgag gcctggttcc ctgaggacca gcgggtactg acccccaacc 1681 tggtggccgc cctgcccccc agcacccatg gggcaggttg gcagctgttt tgcaggactg 1741 tatggtcagc acactcgggg cctacacgga tggccacagc cgtcgcccgc tgcgccccag 1801 atgaggagct gctgagctgc tccagtttct ccaggagtgg gaagcggcgg ggcgagcgca 1861 tggaggccca agggggcaag ctggtctgcc gggcccacaa cgcttttggg ggtgagggtg 1921 tctacgccat tgccaggtgc tgcctgctac cccaggccaa ctgcagcgtc cacacagctc 1981 caccagctga ggccagcatg gggacccgtg tccactgcca ccaacagggc cacgtcctca 2041 caggctgcag ctcccactgg gaggtggagg accttggcac ccacaagccg cctgtgctga 2101 ggccacgagg tcagcccaac cagtgcgtgg gccacaggga ggccagcatc cacgcttcct 2161 gctgccatgc cccaggtctg gaatgcaaag tcaaggagca tggaatcccg gcccctcagg 2221 agcaggtgac cgtggcctgc gaggagggct ggaccctgac tggctgcagt gccctccctg 2281 ggacctccca cgtcctgggg gcctacgccg tagacaacac gtgtgtagtc aggagccggg 2341 acgtcagcac tacaggcagc accagcgaag gggccgtgac agccgttgcc atctgctgcc 2401 ggagccggca cctggcgcag gcctcccagg agctccagtg acagccccat cccaggatgg 2461 gtgtctgggg agggtcaagg gctggggctg agctttaaaa tggttccgac ttgtccctctPAT059857-PCT-SEC012521 ctcagccctc catggcctgg cacgagggga tggggatgct tccgcctttc cggggctgct 2581 ggcctggccc ttgagtgggg cagcctcctt gcctggaact cactcactct gggtgcctcc 2641 tccccaggtg gaggtgccag gaagctccct ccctcactgt ggggcatttc accattcaaa 2701 caggtcgagc tgtgctcggg tgctgccagc tgctcccaat gtgccgatgt ccgtgggcag 2761 aatgactttt attgagctct tgttccgtgc caggcattca atcctcaggt ctccaccaag 2821 gaggcaggat tcttcccatg gataggggag ggggcggtag gggctgcagg gacaaacatc 2881 gttggggggt gagtgtgaaa ggtgctgatg gccctcatct ccagctaact gtggagaagc 2941 ccctgggggc tccctgatta atggaggctt agctttctgg atggcatcta gccagaggct3001 ggagacaggt gcgcccctgg tggtcacagg ctgtgccttg gtttcctgag ccacctttac3061 tctgctctat gccaggctgt gctagcaaca cccaaaggtg gcctgcgggg agccatcacc 3121 taggactgac tcggcagtgt gcagtggtgc atgcactgtc tcagccaacc cgctccacta 3181 cccggcaggg tacacattcg cacccctact tcacagagga agaaacctgg aaccagaggg 3241 ggcgtgcctg ccaagctcac acagcaggaa ctgagccaga aacgcagatt gggctggctc 3301 tgaagccaag cctcttctta cttcacccgg ctgggctcct catttttacg ggtaacagtg 3361 aggctgggaa ggggaacaca gaccaggaag ctcggtgagt gatggcagaa cgatgcctgc 3421 aggcatggaa ctttttccgt tatcacccag gcctgattca ctggcctggc ggagatgctt 3481 ctaaggcatg gtcgggggag agggccaaca actgtccctc cttgagcacc agccccaccc 3541 aagcaagcag acatttatct tttgggtctg tcctctctgt tgccttttta cagccaactt 3601 ttctagacct gttttgcttt tgtaacttga agatatttat tctgggtttt gtagcatttt3661 tattaatatg gtgacttttt aaaataaaaa caaacaaacg ttgtcctaac aaaaaaaaaa 3721 aaaaaaaaaa a

[0141] Homo sapiens PCSK9, transcript variant 1, mRNA: GenBank: NM_174936.4 (SEQ ID NO: 2)1 agcgacgtcg aggcgctcat ggttgcaggc gggcgccgcc gttcagttca gggtctgagc61 ctggaggagt gagccaggca gtgagactgg ctcgggcggg ccgggacgcg tcgttgcagc 121 agcggctccc agctcccagc caggattccg cgcgcccctt cacgcgccct gctcctgaac181 ttcagctcct gcacagtcct ccccaccgca aggctcaagg cgccgccggc gtggaccgcg 241 cacggcctct aggtctcctc gccaggacag caacctctcc cctggccctc atgggcaccg301 tcagctccag gcggtcctgg tggccgctgc cactgctgct gctgctgctg ctgctcctgg361 gtcccgcggg cgcccgtgcg caggaggacg aggacggcga ctacgaggag ctggtgctagPAT059857-PCT-SEC01421 ccttgcgttc cgaggaggac ggcctggccg aagcacccga gcacggaacc acagccacct 481 tccaccgctg cgccaaggat ccgtggaggt tgcctggcac ctacgtggtg gtgctgaagg541 aggagaccca cctctcgcag tcagagcgca ctgcccgccg cctgcaggcc caggctgccc 601 gccggggata cctcaccaag atcctgcatg tcttccatgg ccttcttcct ggcttcctgg661 tgaagatgag tggcgacctg ctggagctgg ccttgaagtt gccccatgtc gactacatcg721 aggaggactc ctctgtcttt gcccagagca tcccgtggaa cctggagcgg attacccctc781 cacggtaccg ggcggatgaa taccagcccc ccgacggagg cagcctggtg gaggtgtatc 841 tcctagacac cagcatacag agtgaccacc gggaaatcga gggcagggtc atggtcaccg 901 acttcgagaa tgtgcccgag gaggacggga cccgcttcca cagacaggcc agcaagtgtg 961 acagtcatgg cacccacctg gcaggggtgg tcagcggccg ggatgccggc gtggccaagg 1021 gtgccagcat gcgcagcctg cgcgtgctca actgccaagg gaagggcacg gttagcggca 1081 ccctcatagg cctggagttt attcggaaaa gccagctggt ccagcctgtg gggccactgg1141 tggtgctgct gcccctggcg ggtgggtaca gccgcgtcct caacgccgcc tgccagcgcc 1201 tggcgagggc tggggtcgtg ctggtcaccg ctgccggcaa cttccgggac gatgcctgcc 1261 tctactcccc agcctcagct cccgaggtca tcacagttgg ggccaccaat gcccaagacc 1321 agccggtgac cctggggact ttggggacca actttggccg ctgtgtggac ctctttgccc1381 caggggagga catcattggt gcctccagcg actgcagcac ctgctttgtg tcacagagtg1441 ggacatcaca ggctgctgcc cacgtggctg gcattgcagc catgatgctg tctgccgagc 1501 cggagctcac cctggccgag ttgaggcaga gactgatcca cttctctgcc aaagatgtca 1561 tcaatgaggc ctggttccct gaggaccagc gggtactgac ccccaacctg gtggccgccc 1621 tgccccccag cacccatggg gcaggttggc agctgttttg caggactgta tggtcagcac 1681 actcggggcc tacacggatg gccacagccg tcgcccgctg cgccccagat gaggagctgc 1741 tgagctgctc cagtttctcc aggagtggga agcggcgggg cgagcgcatg gaggcccaag 1801 ggggcaagct ggtctgccgg gcccacaacg cttttggggg tgagggtgtc tacgccattg 1861 ccaggtgctg cctgctaccc caggccaact gcagcgtcca cacagctcca ccagctgagg 1921 ccagcatggg gacccgtgtc cactgccacc aacagggcca cgtcctcaca ggctgcagct 1981 cccactggga ggtggaggac cttggcaccc acaagccgcc tgtgctgagg ccacgaggtc 2041 agcccaacca gtgcgtgggc cacagggagg ccagcatcca cgcttcctgc tgccatgccc 2101 caggtctgga atgcaaagtc aaggagcatg gaatcccggc ccctcaggag caggtgaccg 2161 tggcctgcga ggagggctgg accctgactg gctgcagtgc cctccctggg acctcccacgPAT059857-PCT-SEC012221 tcctgggggc ctacgccgta gacaacacgt gtgtagtcag gagccgggac gtcagcacta 2281 caggcagcac cagcgaaggg gccgtgacag ccgttgccat ctgctgccgg agccggcacc 2341 tggcgcaggc ctcccaggag ctccagtgac agccccatcc caggatgggt gtctggggag 2401 ggtcaagggc tggggctgag ctttaaaatg gttccgactt gtccctctct cagccctcca 2461 tggcctggca cgaggggatg gggatgcttc cgcctttccg gggctgctgg cctggccctt 2521 gagtggggca gcctccttgc ctggaactca ctcactctgg gtgcctcctc cccaggtgga 2581 ggtgccagga agctccctcc ctcactgtgg ggcatttcac cattcaaaca ggtcgagctg2641 tgctcgggtg ctgccagctg ctcccaatgt gccgatgtcc gtgggcagaa tgacttttat 2701 tgagctcttg ttccgtgcca ggcattcaat cctcaggtct ccaccaagga ggcaggattc 2761 ttcccatgga taggggaggg ggcggtaggg gctgcaggga caaacatcgt tggggggtga 2821 gtgtgaaagg tgctgatggc cctcatctcc agctaactgt ggagaagccc ctgggggctc 2881 cctgattaat ggaggcttag ctttctggat ggcatctagc cagaggctgg agacaggtgc2941 gcccctggtg gtcacaggct gtgccttggt ttcctgagcc acctttactc tgctctatgc 3001 caggctgtgc tagcaacacc caaaggtggc ctgcggggag ccatcaccta ggactgactc 3061 ggcagtgtgc agtggtgcat gcactgtctc agccaacccg ctccactacc cggcagggta 3121 cacattcgca cccctacttc acagaggaag aaacctggaa ccagaggggg cgtgcctgcc 3181 aagctcacac agcaggaact gagccagaaa cgcagattgg gctggctctg aagccaagcc 3241 tcttcttact tcacccggct gggctcctca tttttacggg taacagtgag gctgggaagg 3301 ggaacacaga ccaggaagct cggtgagtga tggcagaacg atgcctgcag gcatggaact 3361 ttttccgtta tcacccaggc ctgattcact ggcctggcgg agatgcttct aaggcatggt 3421 cgggggagag ggccaacaac tgtccctcct tgagcaccag ccccacccaa gcaagcagac 3481 atttatcttt tgggtctgtc ctctctgttg cctttttaca gccaactttt ctagacctgt3541 tttgcttttg taacttgaag atatttattc tgggttttgt agcattttta ttaatatggt3601 gactttttaa aataaaaaca aacaaacgtt gtcctaa

[0142] In certain aspects, PCSK9 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 PCSK9 gene (mRNA) or protein sequences. In certain aspects, PCSK9 may include a fragment, variant, or mutant of the protein having the same or in similar in vivo or in vitroPAT059857-PCT-SEC01enzymatic (e.g., reductase) activity, for example, having about 80%, 85%, 90%, 95%, or 99% or the native protein activity, to bind to LDLR on the surface of a liver cell.

[0143] The term “Compound” as used herein refers to a double stranded RNA (e.g., PCSK9 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. In the present disclosure, Compounds are numbered in examples, e.g., “DI”

[0144] 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., PCSK9 protein) or an mRNA target (e.g., PCSK9 mRNA). In certain aspects, inhibition refers to a reduction of amount of a target biomolecule (e.g., PCSK9 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., PCSK9 protein or mRNA) from an indirect interaction (e.g., inhibiting or regulating other transcriptional or translational factors).

[0145] 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.PAT059857-PCT-SEC01

[0146] 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., PCSK9), as manifested by a reduction of the amount of the target gene mRNA (e.g., PCSK9 mRNA) or protein (e.g., PCSK9) 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”).

[0147] In some embodiments, the level or expression of the target gene (e.g., PCSK9) 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., PCSK9) 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:(mRNA in control cells) - (mRNA in treated cells)- • 100 %(mRNA in control cells)

[0148] 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., PCSK9) expression, e.g., the amount of protein encoded by a target gene (e.g., PCSK9), alteration in expression of the protein whose expression is dependent on the target gene (e.g., PCSK9), alteration in an activity of the enzyme (e.g., PCSK9) encoded by the target gene (e.g., PCSK9). In some embodiments, the level or expression of the protein (e.g., PCSK9) 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.

[0149] As used herein, the term “down-regulate” or “down-regulating” refers to any decrease (e.g., statistically significant) in a biological activity and / or expression of the target protein (e.g., PCSK9), 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., PCSK9) level, activity and / or expression.PAT059857-PCT-SEC01

[0150] 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 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.

[0151] 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, camphorsulf onate, caprate, chloride / hydrochloride, chlortheophyllonate, citrate, ethandisulfonate, fumarate, gluceptate, gluconate, glucuronate, glutamate, glutarate, glycolate, hippurate, hydroiodide / iodide, isethionate,PAT059857-PCT-SEC01lactate, 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.

[0152] 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).

[0153] 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.

[0154] 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.

[0155] 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.

[0156] 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 PCSK9 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.,PAT059857-PCT-SEC01PCSK9 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.

[0157] 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. In some embodiments, hypertriglyceridemia means 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. In some embodiments, hypercholesterolemia means at least about 240 mg / dL of total cholesterol.

[0158] 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.

[0159] 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.

[0160] 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.

[0161] 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 medicalPAT059857-PCT-SEC01response 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., PCSK9), or (ii) associated with its activity, or (iii) characterized by activity (normal or abnormal) of the protein encoded by the target gene (e.g., PCSK9); or (2) reduce or inhibit the activity of the protein encoded by the target gene (e.g., PCSK9); or (3) reduce or inhibit the expression of the target gene (e.g., PCSK9). 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., PCSK9); or at least partially reducing or inhibiting the expression of the protein (e.g., PCSK9) 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., PCSK9 or other proteins relevant to cholesterol uptake).

[0162] 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. 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 about 5%, about 10%, about 15%, about 20%, about 25%, about 40%, about 50%, about 60%, about 75%, about 80%, about 90%, or about 95%. Therapeutic efficacy can also be expressed as “-fold” increase or decrease. ForPAT059857-PCT-SEC01example, 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.

[0163] 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., PCSK9) in the absence of RNAi agents as described herein.

[0164] 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.

[0165] 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., Ci-Cio means one to ten carbons). Examples of alkyl include, but are not limited to, groups such as C1-30 alkyl, C1-25 alkyl, C1-20 alkyl, C1-15 alkyl, C1-12 alkyl, Ci-10 alkyl, C1-8 alkyl, C1-6 alkyl, Ci-4 alkyl, or C1-3 alkyl. 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.

[0166] 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-.

[0167] 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 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 Ci-30 alkenyl, C1-25 alkenyl, C1-20 alkenyl, C1-15 alkenyl, C1-12 alkenyl, C1-10 alkenyl, C1-8 alkenyl, C1-6 alkenyl, Ci-4 alkenyl, or C1-3 alkenyl. For example, C2-6 alkenyl include, but are not limited to,PAT059857-PCT-SEC01ethenyl (vinyl), prop-l-enyl, but-l-enyl, pent-l-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-.

[0168] 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-C10 means one to ten carbons). Examples of alkynyl include, but are not limited to, groups such as C1-30 alkynyl, Ci-25 alkynyl, C1-20 alkynyl, C1-15 alkynyl, C1-12 alkynyl, C1-10 alkynyl, C1-8 alkynyl, C1-6 alkynyl, Ci-4 alkynyl, or C1-3 alkynyl. 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-.

[0169] As used herein, the term “alkoxy” refers to a radical of the formula -ORawhere Rais an alkyl (e.g., C1-30 alkyl, Ci-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 generally defined above. For example, C1-6 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, pentoxy, and hexoxy.

[0170] As used herein, the term " alkoxyalkyl " refers to a radical of the formula -Ra-0-Rbwhere each Raand Rbis independently an alkyl (e.g., C1-30 alkyl, Ci-25 alkyl, C1-20 alkyl, C1-15 alkyl, Ci-i2alkyl, Ci-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, Ci-ealkoxy Ci-ealkyl include, but are not limited to, methoxy-methyl, methoxy-ethyl, ethoxy-ethyl, 1 -ethoxy-propyl and 2-methoxy-butyl.

[0171] As used herein, the term “alkylcarbonyl” refers to a radical of the formula -C(=O)-Ra where Rais an alkyl (e.g., C1-30 alkyl, Ci-25 alkyl, C1-20 alkyl, C1-15 alkyl, C1-12 alkyl, C1-10 alkyl, Ci-8 alkyl, C1-6 alkyl, Ci-4 alkyl, or C1-3 alkyl) radical as defined above.PAT059857-PCT-SEC01

[0172] As used herein, the term "alkyl-carbonyl alkyl” refers to a radical of the formula, e.g., -Ra-C(=O)-Rbwhere each Raand Rbis independently an alkyl (e.g., C1-30 alkyl, C1-25 alkyl, C1-20 alkyl, Ci-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. The carbon atom of the carbonyl group may be bonded to any carbon atom in either alkyl radical.

[0173] As used herein, the term "alkylaminocarbonyl" refers to a radical of the formula -C(=O)-NH-Rawhere Rais an alkyl (e.g., C1-30 alkyl, C1-25 alkyl, C1-20 alkyl, C1-15 alkyl, C1-12 alkyl, Ci-10 alkyl, Ci-8 alkyl, C1-6 alkyl, Ci-4 alkyl, or C1-3 alkyl) as defined above.

[0174] As used herein, the term "alkoxy carbonyl” 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, C 1-15 alkyl, C1-12 alkyl, C1-10 alkyl, Ci-8 alkyl, C1-6 alkyl, Ci-4 alkyl, or C1-3 alkyl) radical as defined above.

[0175] 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, C1-20 alkyl, Ci-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.

[0176] As used herein, the term "haloalkyl" refers to an alkyl (e.g., C1-30 alkyl, C1-25 alkyl, Ci-20 alkyl, Ci -15 alkyl, Ci -12 alkyl, Ci-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 halogenCi-ealkyl include, but are not limited to, trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, l,3-dibromopropan-2-yl, 3-bromo-2-fluoropropyl and 1,4,4-trifluorobutan-2-yl.

[0177] As used herein, the term "hydroxyalkyl” refers to an alkyl (e.g., C1-30 alkyl, C1-25 alkyl, C1-20 alkyl, Ci-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 hydroxyCi-6 alkyl include, but are not limited to, hydroxy-methyl, 2-hydroxy-ethyl, 2-hydroxy-propyl, 3-hydroxy-propyl and 5-hydroxy-pentyl.

[0178] As used herein, the term “aminoalkyl” refers to an alkyl (e.g., C1-30 alkyl, C1-25 alkyl, C1-20 alkyl, Ci-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 Ci -ealkyl group is replaced by aPAT059857-PCT-SEC01primary amino group. Examples of amino Ci-6 alkyl include, but are not limited to, amino-methyl, 2-amino-ethyl, 2-amino-propyl, 3 -amino-propyl, 3 -amino-pentyl and 5-amino-pentyl.

[0179] As used herein, the term “alkylamino” refers to a radical of the formula -NH-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.

[0180] 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).

[0181] 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).

[0182] 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).

[0183] 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 onePAT059857-PCT-SEC01triple bond between carbon and carbon) and include mono-, di- and multivalent radicals. As used herein, the alkynyl is an uncyclized chain. The heteroalkynyl 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).

[0184] 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-.

[0185] 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, l-(l,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.

[0186] 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, aPAT059857-PCT-SEC016,6-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 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.

[0187] 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 “haloalky 1” are meant to include monohaloalkyl and polyhaloalkyl. For example, the term “halo(Ci-C4)alkyl” includes, but is not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3 -bromopropyl, and the like.

[0188] The symbol“ ” denotes the point of attachment of a chemical moiety to the remainder of a molecule or chemical formula.

[0189] The term “oxo,” as used herein, means an oxygen that is double-bonded to a carbon atom.

[0190] 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 asPAT059857-PCT-SEC01alkylene, 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', =0, =NR', =N— OR', —NR'R", —SR', -halogen, — SiR'R" R'", — 0C(0)R', — C(0)R', — CO2R', — CONR'R", — 0C(0)NR'R", — NR''C(0)R', — NR'— C(0)NR" R'", — NR''C(0)2R', — NR— C(NR'R''R'")=NR"", — NR— C(NR'R")=NR'", — S(O)R', — S(O)2R', — S(0)2NR'R", — NRS02R', — NR'NR" R'", — ONR'R", — NR'C(O)NR''NR'" R'''', — CN, — N02, — NR'S02R", — NR'C(0)R", — NR'C(O)— OR", — NR'OR", in a number ranging from zero to (2m'+l), 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 haloalky 1 (e.g., -CF3and -CH2CF3) and acyl (e.g., -C(0)CH3, — C(O)CF3, — C(O)CH2OCH3, and the like).

[0191] 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.PAT059857-PCT-SEC01

[0192] 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.RNAi Agents

[0193] 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.

[0194] A dsRNA is a complex of ribonucleic acid (RNA) molecules formed in a duplex structure. In certain aspects, the dsRNA may be a small 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 an 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 a sense strand.PCSK9 siRNA (double stranded RNA)

[0195] In an aspect, the disclosure provides a dsRNA interference (dsRNAi) agent that is capable of interacting or recruiting a target mRNA sequence, e.g., PCSK9 mRNA sequence, in the RISC thereby cleaving the target mRNA. The dsRNAi agent can silence PCSK9 gene, e.g., by inhibiting, downregulating, or suppressing the expression of PCSK9 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 PCSK9 expressionPAT059857-PCT-SEC01compared 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 PCSK9 may be demonstrated by a reduction of the amount of a total PCSK9 mRNA in a cell. In some embodiments, the level of silencing the PCSK9 may be demonstrated by a reduction of the amount of a total PCSK9 protein in a cell.

[0196] In some embodiments, expression of the PCSK9 gene (e.g., human PCSK9) 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 PCSK9 gene in untreated cell or subject. In some embodiments, expression of the PCSK9 gene (e.g., human PCSK9) is inhibited by at least about 20% based on the expression level of the PCSK9 gene in untreated cell or subject. In some embodiments, expression of the PCSK9 gene (e.g., human PCSK9) is inhibited by at least about 30% based on the expression level of the PCSK9 gene in untreated cell or subject. In some embodiments, expression of the PCSK9 gene (e.g., human PCSK9) is inhibited by at least about 40% based on the expression level of the PCSK9 gene in untreated cell or subject. In some embodiments, expression of the PCSK9 gene (e.g., human PCSK9) is inhibited by at least about 50% based on the expression level of the PCSK9 gene in untreated cell or subject. In some embodiments, expression of the PCSK9 gene (e.g., human PCSK9) is inhibited by at least about 60% based on the expression level of the PCSK9 gene in untreated cell or subject. In some embodiments, expression of the PCSK9 gene (e.g., human PCSK9) is inhibited by at least about 70% based on the expression level of the PCSK9 gene in untreated cell or subject.

[0197] In some embodiments, inhibition of the expression of the PCSK9 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 PCSK9 (e.g., human PCSK9) may be presented as a percentage of remaining mRNAPAT059857-PCT-SEC01in 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)- • 1 uu %(mRNA in control cells)

[0198] In some embodiments, the level of gene-silencing (e.g., inhibiting, downregulating, or suppressing of the gene) of the PCSK9 (e.g., human PCSK9) may be assessed by measuring a parameter or biomarker, e.g., human PCSK9 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 genesilencing (e.g., inhibiting, downregulating, or suppressing of the gene) of the PCSK9 (e.g., human PCSK9) may be observed or assessed by in a liver (tissue) biopsy of the treated subject.

[0199] 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 (NRC, R being an alkyl group or an aryl group as described herein), or copper (Cu2+).

[0200] In an aspect, the disclosure provides a dsRNA having sequences (e.g., antisense strand sequence) that can recognize a specific region of a PCSK9 mRNA (e.g., human PCSK9 mRNA)PAT059857-PCT-SEC01and lead cleavage of the PCSK9 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.

[0201] 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.

[0202] 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.

[0203] 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 somePAT059857-PCT-SEC01embodiments, the sense strand is 23 nucleotides in length and the antisense strand is 25 nucleotides in length.

[0204] 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 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 hasnucleotide 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 hasnucleotide 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 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.

[0205] 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.PAT059857-PCT-SEC01

[0206] 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 to 6 nucleotides in length, for example, 1 nucleotide, 2 nucleotides, 3 nucleotides, 4 nucleotides, 5 nucleotides, or 6 nucleotides in length.

[0207] 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 some embodiments, the antisense contains two nucleotides overhang at the 3' end and one-nucleotide overhang at the 5' end.

[0208] 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.PAT059857-PCT-SEC01

[0209] In certain aspects, the target PCSK9 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 certain aspects, the target PCSK9 mRNA sequence may range from 19 to 25 nucleotides, from 19 to 23 nucleotides, or from 19 to 21 nucleotides. In some embodiments, the target PCSK9 mRNA sequence may have 15 nucleotides in length. In some embodiments, the target PCSK9 mRNA sequence may have 16 nucleotides in length. In some embodiments, the target PCSK9 mRNA sequence may have 17 nucleotides in length. In some embodiments, the target PCSK9 mRNA sequence may have 18 nucleotides in length. In some embodiments, the target PCSK9 mRNA sequence may have 19 nucleotides in length. In some embodiments, the target PCSK9 mRNA sequence may have 20 nucleotides in length. In some embodiments, the target PCSK9 mRNA sequence may have 21 nucleotides in length. In some embodiments, the target PCSK9 mRNA sequence may have 22 nucleotides in length. In some embodiments, the target PCSK9 mRNA sequence may have 23 nucleotides in length.

[0210] In certain aspects, exemplary dsRNA sequences including sense strands and antisense strands targeting human PCSK9 mRNAs of SEQ ID NO: 2 (GenBank: NM_174936.4) are in TableTable 1: Examples of unmodified nucleotide sequences of PCSK9 siRNAPCS Site Regi Sense sequence SE Antisense sequence SE K9 of on Q Q siRN mRN ID ID A A N NTarg O: O: et*U1 27 5' GUCGAGGCGCUCAUG 3 UGCAACCAUGAGCGCCU 38UTR GUUGCA CGACGU 2 U2 30 5' GAGGCGCUCAUGGUU 4 UCCUGCAACCAUGAGCG 38UTR GCAGGA CCUCGA 3 U3 117 5' CGGGCCGGGACGCGU 5 UCAACGACGCGUCCCGG 38UTR CGUUGA CCCGCC 4 U4 121 5' CCGGGACGCGUCGUU 6 UGCUGCAACGACGCGUC 38UTR GCAGCA CCGGCC 5 U5 153 5' CUCCCAGCCAGGAUU 7 UCGCGGAAUCCUGGCUG 38UTR CCGCGA GGAGCU 6 U6 155 5' CCCAGCCAGGAUUCC 8 UCGCGCGGAAUCCUGGC 38UTR GCGCGA UGGGAG 7PAT059857-PCT-SEC01U7 161 5' CAGGAUUCCGCGCGC 9 UAAGGGGCGCGCGGAAU 38UTR CCCUUA CCUGGC 8 U8 219 5' CUCCCCACCGCAAGGC 10 UUUGAGCCUUGCGGUGG 38UTR UCAAA GGAGGA 9 U9 220 5' UCCCCACCGCAAGGCU 11 UCUUGAGCCUUGCGGUG 39UTR CAAGG GGGAGG 0 U10 256 5' CCGCGCACGGCCUCUA 12 AGACCUAGAGGCCGUGC 39UTR GGUCU GCGGUC 1 Ull 256 5' CCGCGCACGGCCUCUA 13 UGACCUAGAGGCCGUGC 39UTR GGUCA GCGGUC 2 U12 257 5' CGCGCACGGCCUCUA 14 UAGACCUAGAGGCCGUG 39UTR GGUCUA CGCGGU 3 U13 260 5' GCACGGCCUCUAGGU 15 UAGGAGACCUAGAGGCC 39UTR CUCCUA GUGCGC 4 U14 316 CDS CACCGUCAGCUCCAG 16 UACCGCCUGGAGCUGAC 39GCGGUA GGUGCC 5 U15 317 CDS ACCGUCAGCUCCAGG 17 UGACCGCCUGGAGCUGA 39CGGUCC CGGUGC 6 U16 318 CDS CCGUCAGCUCCAGGC 18 AGGACCGCCUGGAGCUG 39GGUCCU ACGGUG 7 U17 318 CDS CCGUCAGCUCCAGGC 19 UGGACCGCCUGGAGCUG 39GGUCCA ACGGUG 8 U18 378 CDS UGGGUCCCGCGGGCG 20 UACGGGCGCCCGCGGGA 39CCCGUG CCCAGG 9 U19 386 CDS GCGGGCGCCCGUGCG 21 UUCCUGCGCACGGGCGC 40CAGGAA CCGCGG 0 U20 392 CDS GCCCGUGCGCAGGAG 22 UUCGUCCUCCUGCGCAC 40GACGAA GGGCGC 1 U21 393 CDS CCCGUGCGCAGGAGG 23 UCUCGUCCUCCUGCGCA 40ACGAGA CGGGCG 2 U22 404 CDS GAGGACGAGGACGGC 24 UUAGUCGCCGUCCUCGU 40GACUAA CCUCCU 3 U23 421 CDS CUACGAGGAGCUGGU 25 UCUAGCACCAGCUCCUC 40GCUAGA GUAGUC 4 U24 425 CDS GAGGAGCUGGUGCUA 26 UAAGGCUAGCACCAGCU 40GCCUUA CCUCGU 5 U25 430 CDS GCUGGUGCUAGCCUU 27 UAACGCAAGGCUAGCAC 40GCGUUA CAGCUC 6 U26 432 CDS UGGUGCUAGCCUUGC 28 UGGAACGCAAGGCUAGC 40GUUCCG ACCAGC 7 U27 437 CDS CUAGCCUUGCGUUCC 29 UUCCUCGGAACGCAAGG 40GAGGAA CUAGCA 8 U28 476 CDS CCCGAGCACGGAACC 30 UGCUGUGGUUCCGUGCU 40ACAGCA CGGGUG 9PAT059857-PCT-SEC01U29 479 CDS GAGCACGGAACCACA 31 UGUGGCUGUGGUUCCGU 41GCCACA GCUCGG 0 U30 490 CDS CACAGCCACCUUCCAC 32 UAGCGGUGGAAGGUGG 41CGCUA CUGUGGU 1 U31 496 CDS CACCUUCCACCGCUGC 33 UUGGCGCAGCGGUGGAA 41GCCAA GGUGGC 2 U32 497 CDS ACCUUCCACCGCUGCG 34 UUUGGCGCAGCGGUGGA 41CCAAG AGGUGG 3 U33 500 CDS UUCCACCGCUGCGCCA 35 AUCCUUGGCGCAGCGGU 41AGGAU GGAAGG 4 U34 500 CDS UUCCACCGCUGCGCCA 36 UUCCUUGGCGCAGCGGU 41AGGAA GGAAGG 5 U35 502 CDS CCACCGCUGCGCCAAG 37 UGAUCCUUGGCGCAGCG 41GAUCA GUGGAA 6 U36 503 CDS CACCGCUGCGCCAAG 38 UGGAUCCUUGGCGCAGC 41GAUCCA GGUGGA 7 U37 505 CDS CCGCUGCGCCAAGGA 39 UACGGAUCCUUGGCGCA 41UCCGUA GCGGUG 8 U38 506 CDS CGCUGCGCCAAGGAU 40 UCACGGAUCCUUGGCGC 41CCGUGA AGCGGU 9 U39 539 CDS ACCUACGUGGUGGUG 41 UUUCAGCACCACCACGU 42CUGAAG AGGUGC 0 U40 570 CDS ACCUCUCGCAGUCAG 42 UGCGCUCUGACUGCGAG 42AGCGCA AGGUGG 1 U41 571 CDS CCUCUCGCAGUCAGA 43 UUGCGCUCUGACUGCGA 42GCGCAA GAGGUG 2 U42 573 CDS UCUCGCAGUCAGAGC 44 UAGUGCGCUCUGACUGC 42GCACUG GAGAGG 3 U43 611 CDS CAGGCUGCCCGCCGG 45 UUAUCCCCGGCGGGCAG 42GGAUAA CCUGGG 4 U44 613 CDS GGCUGCCCGCCGGGG 46 AGGUAUCCCCGGCGGGC 42AUACCU AGCCUG 5 U45 613 CDS GGCUGCCCGCCGGGG 47 UGGUAUCCCCGGCGGGC 42AUACCA AGCCUG 6 U46 616 CDS UGCCCGCCGGGGAUA 48 UUGAGGUAUCCCCGGCG 42CCUCAC GGCAGC 7 U47 620 CDS CGCCGGGGAUACCUC 49 UUUGGUGAGGUAUCCCC 42ACCAAA GGCGGG 8 U48 621 CDS GCCGGGGAUACCUCA 50 UCUUGGUGAGGUAUCCC 42CCAAGA CGGCGG 9 U49 623 CDS CGGGGAUACCUCACC 51 UAUCUUGGUGAGGUAU 43AAGAUA CCCCGGC 0 U50 624 CDS GGGGAUACCUCACCA 52 UGAUCUUGGUGAGGUA 43AGAUCA UCCCCGG 1PAT059857-PCT-SEC01U51 668 CDS CCUGGCUUCCUGGUG 53 UAUCUUCACCAGGAAGC 43AAGAUA CAGGAA 2 U52 680 CDS GUGAAGAUGAGUGGC 54 UAGGUCGCCACUCAUCU 43GACCUA UCACCA 3 U53 683 CDS AAGAUGAGUGGCGAC 55 UAGCAGGUCGCCACUCA 43CUGCUG UCUUCA 4 U54 709 CDS GGCCUUGAAGUUGCC 56 ACAUGGGGCAACUUCAA 43CCAUGU GGCCAG 5 U55 709 CDS GGCCUUGAAGUUGCC 57 UCAUGGGGCAACUUCAA 43CCAUGA GGCCAG 6 U56 711 CDS CCUUGAAGUUGCCCC 58 UGACAUGGGGCAACUUC 43AUGUCA AAGGCC 7 U57 712 CDS CUUGAAGUUGCCCCA 59 UCGACAUGGGGCAACUU 43UGUCGA CAAGGC 8 U58 713 CDS UUGAAGUUGCCCCAU 60 UUCGACAUGGGGCAACU 43GUCGAC UCAAGG 9 U59 714 CDS UGAAGUUGCCCCAUG 61 AGUCGACAUGGGGCAAC 44UCGACU UUCAAG 0 U60 714 CDS UGAAGUUGCCCCAUG 62 UGUCGACAUGGGGCAAC 44UCGACA UUCAAG 1 U61 715 CDS GAAGUUGCCCCAUGU 63 UAGUCGACAUGGGGCAA 44CGACUA CUUCAA 2 U62 716 CDS AAGUUGCCCCAUGUC 64 UUAGUCGACAUGGGGCA 44GACUAC ACUUCA 3 U63 717 CDS AGUUGCCCCAUGUCG 65 UGUAGUCGACAUGGGGC 44ACUACA AACUUC 4 U64 718 CDS GUUGCCCCAUGUCGA 66 AUGUAGUCGACAUGGG 44CUACAU GCAACUU 5 U65 718 CDS GUUGCCCCAUGUCGA 67 UUGUAGUCGACAUGGG 44CUACAA GCAACUU 6 U66 720 CDS UGCCCCAUGUCGACU 68 UGAUGUAGUCGACAUG 44ACAUCG GGGCAAC 7 U67 721 CDS GCCCCAUGUCGACUA 69 UCGAUGUAGUCGACAUG 44CAUCGA GGGCAA 8 U68 722 CDS CCCCAUGUCGACUAC 70 UUCGAUGUAGUCGACAU 44AUCGAA GGGGCA 9 U69 725 CDS CAUGUCGACUACAUC 71 UUCCUCGAUGUAGUCGA 45GAGGAA CAUGGG 0 U70 726 CDS AUGUCGACUACAUCG 72 UCUCCUCGAUGUAGUCG 45AGGAGG ACAUGG 1 U71 728 CDS GUCGACUACAUCGAG 73 UUCCUCCUCGAUGUAGU 45GAGGAA CGACAU 2 U72 731 CDS GACUACAUCGAGGAG 74 UGAGUCCUCCUCGAUGU 45GACUCA AGUCGA 3PAT059857-PCT-SEC01U73 770 CDS AUCCCGUGGAACCUG 75 UCGCUCCAGGUUCCACG 45GAGCGG GGAUGC 4 U74 794 CDS ACCCCUCCACGGUACC 76 UGCCCGGUACCGUGGAG 45GGGCG GGGUAA 5 U75 795 CDS CCCCUCCACGGUACCG 77 UCGCCCGGUACCGUGGA 45GGCGA GGGGUA 6 U76 796 CDS CCCUCCACGGUACCGG 78 UCCGCCCGGUACCGUGG 45GCGGA AGGGGU 7 U77 798 CDS CUCCACGGUACCGGG 79 UAUCCGCCCGGUACCGU 45CGGAUA GGAGGG 8 U78 799 CDS UCCACGGUACCGGGC 80 UCAUCCGCCCGGUACCG 45GGAUGA UGGAGG 9 U79 800 CDS CCACGGUACCGGGCG 81 UUCAUCCGCCCGGUACC 46GAUGAA GUGGAG 0 U80 801 CDS CACGGUACCGGGCGG 82 AUUCAUCCGCCCGGUAC 46AUGAAU CGUGGA 1 U81 801 CDS CACGGUACCGGGCGG 83 UUUCAUCCGCCCGGUAC 46AUGAAA CGUGGA 2 U82 802 CDS ACGGUACCGGGCGGA 84 UAUUCAUCCGCCCGGUA 46UGAAUA CCGUGG 3 U83 806 CDS UACCGGGCGGAUGAA 85 UUGGUAUUCAUCCGCCC 46UACCAG GGUACC 4 U84 840 CDS GCAGCCUGGUGGAGG 86 UAUACACCUCCACCAGG 46UGUAUA CUGCCU 5 U85 843 CDS GCCUGGUGGAGGUGU 87 UGAGAUACACCUCCACC 46AUCUCA AGGCUG 6 U86 847 CDS GGUGGAGGUGUAUCU 88 UCUAGGAGAUACACCUC 46CCUAGA CACCAG 7 U87 848 CDS GUGGAGGUGUAUCUC 89 UUCUAGGAGAUACACCU 46CUAGAA CCACCA 8 U88 850 CDS GGAGGUGUAUCUCCU 90 UUGUCUAGGAGAUACAC 46AGACAA CUCCAC 9 U89 851 CDS GAGGUGUAUCUCCUA 91 UGUGUCUAGGAGAUAC 47GACACA ACCUCCA 0 U90 852 CDS AGGUGUAUCUCCUAG 92 UGGUGUCUAGGAGAUA 47ACACCA CACCUCC 1 U91 853 CDS GGUGUAUCUCCUAGA 93 UUGGUGUCUAGGAGAU 47CACCAA ACACCUC 2 U92 856 CDS GUAUCUCCUAGACAC 94 AUGCUGGUGUCUAGGA 47CAGCAU GAUACAC 3 U93 856 CDS GUAUCUCCUAGACAC 95 UUGCUGGUGUCUAGGA 47CAGCAA GAUACAC 4 U94 857 CDS UAUCUCCUAGACACC 96 UAUGCUGGUGUCUAGG 47AGCAUA AGAUACA 5PAT059857-PCT-SEC01U95 858 CDS AUCUCCUAGACACCA 97 UUAUGCUGGUGUCUAG 47GCAUAC GAGAUAC 6 U96 862 CDS CCUAGACACCAGCAU 98 UUCUGUAUGCUGGUGUC 47ACAGAA UAGGAG 7 U97 868 CDS CACCAGCAUACAGAG 99 UGGUCACUCUGUAUGCU 47UGACCA GGUGUC 8 U98 870 CDS CCAGCAUACAGAGUG 10 UGUGGUCACUCUGUAUG 47ACCACA 0 CUGGUG 9 U99 873 CDS GCAUACAGAGUGACC 10 UCCGGUGGUCACUCUGU 48ACCGGA 1 AUGCUG 0 U10 875 CDS AUACAGAGUGACCAC 10 UUCCCGGUGGUCACUCU 48 0 CGGGAA 2 GUAUGC 1 U1O 876 CDS UACAGAGUGACCACC 10 UUUCCCGGUGGUCACUC 48 1 GGGAAA 3 UGUAUG 2 U1O 878 CDS CAGAGUGACCACCGG 10 UAUUUCCCGGUGGUCAC 48 2 GAAAUA 4 UCUGUA 3 U1O 879 CDS AGAGUGACCACCGGG 10 UGAUUUCCCGGUGGUCA 48 3 AAAUCG 5 CUCUGU 4 U1O 880 CDS GAGUGACCACCGGGA 10 UCGAUUUCCCGGUGGUC 48 4 AAUCGA 6 ACUCUG 5 U1O 881 CDS AGUGACCACCGGGAA 10 UUCGAUUUCCCGGUGGU 48 5 AUCGAG 7 CACUCU 6 U1O 882 CDS GUGACCACCGGGAAA 10 UCUCGAUUUCCCGGUGG 48 6 UCGAGA 8 UCACUC 7 U1O 883 CDS UGACCACCGGGAAAU 10 UCCUCGAUUUCCCGGUG 48 7 CGAGGG 9 GUCACU 8 U1O 892 CDS GGAAAUCGAGGGCAG 11 AUGACCCUGCCCUCGAU 48 8 GGUCAU 0 UUCCCG 9 U1O 892 CDS GGAAAUCGAGGGCAG 11 UUGACCCUGCCCUCGAU 49 9 GGUCAA 1 UUCCCG 0 Ull 893 CDS GAAAUCGAGGGCAGG 11 UAUGACCCUGCCCUCGA 49 0 GUCAUA 2 UUUCCC 1 Ull 894 CDS AAAUCGAGGGCAGGG 11 UCAUGACCCUGCCCUCG 49 1 UCAUGG 3 AUUUCC 2 Ull 896 CDS AUCGAGGGCAGGGUC 11 UACCAUGACCCUGCCCU 49 2 AUGGUC 4 CGAUUU 3 Ull 900 CDS AGGGCAGGGUCAUGG 11 UGGUGACCAUGACCCUG 49 3 UCACCG 5 CCCUCG 4 Ull 902 CDS GGCAGGGUC AUGGUC 11 UUCGGUGACCAUGACCC 49 4 ACCGAA 6 UGCCCU 5 Ull 903 CDS GCAGGGUC AUGGUC A 11 AGUCGGUGACCAUGACC 49 5 CCGACU 7 CUGCCC 6 Ull 903 CDS GCAGGGUC AUGGUC A 11 UGUCGGUGACCAUGACC 496 CCGACA 8 CUGCCC 7PAT059857-PCT-SEC01Ull 904 CDS CAGGGUCAUGGUCAC 11 AAGUCGGUGACCAUGAC 49 7 CGACUU 9 CCUGCC 8 Ull 904 CDS CAGGGUCAUGGUCAC 12 UAGUCGGUGACCAUGAC 49 8 CGACUA 0 CCUGCC 9 Ull 945 CDS ACGGGACCCGCUUCC 12 UUCUGUGGAAGCGGGUC 50 9 ACAGAC 1 CCGUCC 0 U12 946 CDS CGGGACCCGCUUCCAC 12 UGUCUGUGGAAGCGGG 50 0 AGACA 2 UCCCGUC 1 U12 960 CDS ACAGACAGGCCAGCA 12 UACACUUGCUGGCCUGU 50 1 AGUGUG 3 CUGUGG 2 U12 976 CDS GUGUGACAGUCAUGG 12 UGGGUGCCAUGACUGUC 50 2 CACCCA 4 ACACUU 3 U12 993 CDS CCCACCUGGCAGGGG 12 UGACCACCCCUGCCAGG 50 3 UGGUCA 5 UGGGUG 4 U12 994 CDS CCACCUGGCAGGGGU 12 UUGACCACCCCUGCCAG 50 4 GGUCAA 6 GUGGGU 5 U12 1054 CDS CAGCCUGCGCGUGCU 12 UAGUUGAGCACGCGCAG 50 5 CAACUA 7 GCUGCG 6 U12 1055 CDS AGCCUGCGCGUGCUC 12 UCAGUUGAGCACGCGCA 50 6 AACUGC 8 GGCUGC 7 U12 1059 CDS UGCGCGUGCUCAACU 12 UUUGGCAGUUGAGCACG 50 7 GCCAAG 9 CGCAGG 8 U12 1060 CDS GCGCGUGCUCAACUG 13 UCUUGGCAGUUGAGCAC 50 8 CCAAGA 0 GCGCAG 9 U12 1070 CDS AACUGCCAAGGGAAG 13 UGUGCCCUUCCCUUGGC 51 9 GGCACG 1 AGUUGA 0 U13 1071 CDS ACUGCCAAGGGAAGG 13 UCGUGCCCUUCCCUUGG 51 0 GCACGG 2 CAGUUG 1 U13 1081 CDS GAAGGGCACGGUUAG 13 UUGCCGCUAACCGUGCC 51 1 CGGCAA 3 CUUCCC 2 U13 1082 CDS AAGGGCACGGUUAGC 13 UGUGCCGCUAACCGUGC 51 2 GGCACC 4 CCUUCC 3 U13 1085 CDS GGCACGGUUAGCGGC 13 UAGGGUGCCGCUAACCG 51 3 ACCCUA 5 UGCCCU 4 U13 1088 CDS ACGGUUAGCGGCACC 13 UAUGAGGGUGCCGCUAA 51 4 CUCAUA 6 CCGUGC 5 U13 1089 CDS CGGUUAGCGGCACCC 13 UUAUGAGGGUGCCGCUA 51 5 UCAUAA 7 ACCGUG 6 U13 1090 CDS GGUUAGCGGCACCCU 13 UCUAUGAGGGUGCCGCU 51 6 CAUAGA 8 AACCGU 7 U13 1097 CDS GGCACCCUCAUAGGC 13 UUCCAGGCCUAUGAGGG 51 7 CUGGAA 9 UGCCGC 8 U13 1099 CDS CACCCUCAUAGGCCU 14 AACUCCAGGCCUAUGAG 518 GGAGUU 0 GGUGCC 9PAT059857-PCT-SEC01U13 1100 CDS ACCCUCAUAGGCCUG 14 AAACUCCAGGCCUAUGA 52 9 GAGUUU 1 GGGUGC 0 U14 1102 CDS CCUCAUAGGCCUGGA 14 AUAAACUCCAGGCCUAU 52 0 GUUUAU 2 GAGGGU 1 U14 1102 CDS CCUCAUAGGCCUGGA 14 UUAAACUCCAGGCCUAU 52 1 GUUUAA 3 GAGGGU 2 U14 1106 CDS AUAGGCCUGGAGUUU 14 UCGAAUAAACUCCAGGC 52 2 AUUCGG 4 CUAUGA 3 U14 1107 CDS UAGGCCUGGAGUUUA 14 UCCGAAUAAACUCCAGG 52 3 UUCGGA 5 CCUAUG 4 U14 1108 CDS AGGCCUGGAGUUUAU 14 UUCCGAAUAAACUCCAG 52 4 UCGGAA 6 GCCUAU 5 U14 1111 CDS CCUGGAGUUUAUUCG 14 UUUUUCCGAAUAAACUC 52 5 GAAAAA 7 CAGGCC 6 U14 1112 CDS CUGGAGUUUAUUCGG 14 UCUUUUCCGAAUAAACU 52 6 AAAAGA 8 CCAGGC 7 U14 1113 CDS UGGAGUUUAUUCGGA 14 UGCUUUUCCGAAUAAAC 52 7 AAAGCC 9 UCCAGG 8 U14 1118 CDS UUUAUUCGGAAAAGC 15 UAGCUGGCUUUUCCGAA 52 8 CAGCUG 0 UAAACU 9 U14 1168 CDS GCUGCCCCUGGCGGG 15 UACCCACCCGCCAGGGG 53 9 UGGGUA 1 CAGCAG 0 U15 1169 CDS CUGCCCCUGGCGGGU 15 UUACCCACCCGCCAGGG 53 0 GGGUAA 2 GCAGCA 1 U15 1170 CDS UGCCCCUGGCGGGUG 15 UGUACCCACCCGCCAGG 53 1 GGUACA 3 GGCAGC 2 U15 1221 CDS UGGCGAGGGCUGGGG 15 UCACGACCCCAGCCCUC 53 2 UCGUGC 4 GCCAGG 3 U15 1228 CDS GGCUGGGGUCGUGCU 15 UUGACCAGCACGACCCC 53 3 GGUCAA 5 AGCCCU 4 U15 1234 CDS GGUCGUGCUGGUCAC 15 UCAGCGGUGACCAGCAC 53 4 CGCUGA 6 GACCCC 5 U15 1237 CDS CGUGCUGGUCACCGC 15 UCGGCAGCGGUGACCAG 53 5 UGCCGA 7 CACGAC 6 U15 1238 CDS GUGCUGGUCACCGCU 15 UCCGGCAGCGGUGACCA 53 6 GCCGGA 8 GCACGA 7 U15 1241 CDS CUGGUCACCGCUGCC 15 UUUGCCGGCAGCGGUGA 53 7 GGCAAA 9 CCAGCA 8 U15 1246 CDS CACCGCUGCCGGCAAC 16 UGGAAGUUGCCGGCAGC 53 8 UUCCA 0 GGUGAC 9 U15 1252 CDS UGCCGGCAACUUCCG 16 UCGUCCCGGAAGUUGCC 54 9 GGACGA 1 GGCAGC 0 U16 1253 CDS GCCGGCAACUUCCGG 16 AUCGUCCCGGAAGUUGC 540 GACGAU 2 CGGCAG 1PAT059857-PCT-SEC01U16 1253 CDS GCCGGCAACUUCCGG 16 UUCGUCCCGGAAGUUGC 54 1 GACGAA 3 CGGCAG 2 U16 1254 CDS CCGGCAACUUCCGGG 16 UAUCGUCCCGGAAGUUG 54 2 ACGAUA 4 CCGGCA 3 U16 1255 CDS CGGCAACUUCCGGGA 16 UCAUCGUCCCGGAAGUU 54 3 CGAUGA 5 GCCGGC 4 U16 1256 CDS GGCAACUUCCGGGAC 16 UGCAUCGUCCCGGAAGU 54 4 GAUGCA 6 UGCCGG 5 U16 1257 CDS GCAACUUCCGGGACG 16 AGGCAUCGUCCCGGAAG 54 5 AUGCCU 7 UUGCCG 6 U16 1257 CDS GCAACUUCCGGGACG 16 UGGCAUCGUCCCGGAAG 54 6 AUGCCA 8 UUGCCG 7 U16 1258 CDS CAACUUCCGGGACGA 16 UAGGCAUCGUCCCGGAA 54 7 UGCCUA 9 GUUGCC 8 U16 1260 CDS ACUUCCGGGACGAUG 17 UGCAGGCAUCGUCCCGG 54 8 CCUGCC 0 AAGUUG 9 U16 1292 CDS GCCUCAGCUCCCGAG 17 UAUGACCUCGGGAGCUG 55 9 GUCAUA 1 AGGCUG 0 U17 1293 CDS CCUCAGCUCCCGAGG 17 UGAUGACCUCGGGAGCU 55 0 UCAUCA 2 GAGGCU 1 U17 1296 CDS CAGCUCCCGAGGUCA 17 UUGUGAUGACCUCGGGA 55 1 UCACAA 3 GCUGAG 2 U17 1299 CDS CUCCCGAGGUCAUCA 17 UAACUGUGAUGACCUCG 55 2 CAGUUA 4 GGAGCU 3 U17 1308 CDS UCAUCACAGUUGGGG 17 UGGUGGCCCCAACUGUG 55 3 CCACCA 5 AUGACC 4 U17 1309 CDS CAUCACAGUUGGGGC 17 UUGGUGGCCCCAACUGU 55 4 CACCAA 6 GAUGAC 5 U17 1354 CDS GGGGACUUUGGGGAC 17 AAGUUGGUCCCCAAAGU 55 5 CAACUU 7 CCCCAG 6 U17 1354 CDS GGGGACUUUGGGGAC 17 UAGUUGGUCCCCAAAGU 55 6 CAACUA 8 CCCCAG 7 U17 1355 CDS GGGACUUUGGGGACC 17 AAAGUUGGUCCCCAAAG 55 7 AACUUU 9 UCCCCA 8 U17 1355 CDS GGGACUUUGGGGACC 18 UAAGUUGGUCCCCAAAG 55 8 AACUUA 0 UCCCCA 9 U17 1357 CDS GACUUUGGGGACCAA 18 UCAAAGUUGGUCCCCAA 56 9 CUUUGA 1 AGUCCC 0 U18 1359 CDS CUUUGGGGACCAACU 18 UGCCAAAGUUGGUCCCC 56 0 UUGGCA 2 AAAGUC 1 U18 1361 CDS UUGGGGACC AACUUU 18 UCGGCCAAAGUUGGUCC 56 1 GGCCGC 3 CCAAAG 2 U18 1365 CDS GGACCAACUUUGGCC 18 UACAGCGGCCAAAGUUG 562 GCUGUA 4 GUCCCC 3PAT059857-PCT-SEC01U18 1367 CDS ACCAACUUUGGCCGC 18 UACACAGCGGCCAAAGU 56 3 UGUGUG 5 UGGUCC 4 U18 1376 CDS GGCCGCUGUGUGGAC 18 AAAGAGGUCCACACAGC 56 4 CUCUUU 6 GGCCAA 5 U18 1376 CDS GGCCGCUGUGUGGAC 18 UAAGAGGUCCACACAGC 56 5 CUCUUA 7 GGCCAA 6 U18 1378 CDS CCGCUGUGUGGACCU 18 UCAAAGAGGUCCACACA 56 6 CUUUGA 8 GCGGCC 7 U18 1379 CDS CGCUGUGUGGACCUC 18 UGCAAAGAGGUCCACAC 56 7 UUUGCA 9 AGCGGC 8 U18 1401 CDS CAGGGGAGGACAUCA 19 UACCAAUGAUGUCCUCC 56 8 UUGGUA 0 CCUGGG 9 U18 1409 CDS GACAUCAUUGGUGCC 19 UCUGGAGGCACCAAUGA 57 9 UCCAGA 1 UGUCCU 0 U19 1414 CDS CAUUGGUGCCUCCAG 19 UAGUCGCUGGAGGCACC 57 0 CGACUA 2 AAUGAU 1 U19 1430 CDS GACUGCAGCACCUGC 19 UACAAAGCAGGUGCUGC 57 1 UUUGUA 3 AGUCGC 2 U19 1433 CDS UGCAGCACCUGCUUU 19 UGACACAAAGCAGGUGC 57 2 GUGUCA 4 UGCAGU 3 U19 1435 CDS CAGCACCUGCUUUGU 19 UGUGACACAAAGCAGGU 57 3 GUCACA 5 GCUGCA 4 U19 1478 CDS GCCCACGUGGCUGGC 19 UGCAAUGCCAGCCACGU 57 4 AUUGCA 6 GGGCAG 5 U19 1488 CDS CUGGCAUUGCAGCCA 19 UCAUCAUGGCUGCAAUG 57 5 UGAUGA 7 CCAGCC 6 U19 1489 CDS UGGCAUUGCAGCCAU 19 AGCAUCAUGGCUGCAAU 57 6 GAUGCU 8 GCCAGC 7 U19 1489 CDS UGGCAUUGCAGCCAU 19 UGCAUCAUGGCUGCAAU 57 7 GAUGCA 9 GCCAGC 8 U19 1494 CDS UUGCAGCCAUGAUGC 20 UAGACAGCAUCAUGGCU 57 8 UGUCUG 0 GCAAUG 9 U19 1502 CDS AUGAUGCUGUCUGCC 20 UGGCUCGGCAGACAGCA 58 9 GAGCCG 1 UCAUGG 0 U20 1504 CDS GAUGCUGUCUGCCGA 20 UCCGGCUCGGCAGACAG 58 0 GCCGGA 2 CAUCAU 1 U20 1505 CDS AUGCUGUCUGCCGAG 20 UUCCGGCUCGGCAGACA 58 1 CCGGAG 3 GCAUCA 2 U20 1522 CDS GGAGCUCACCCUGGC 20 AACUCGGCCAGGGUGAG 58 2 CGAGUU 4 CUCCGG 3 U20 1522 CDS GGAGCUCACCCUGGC 20 UACUCGGCCAGGGUGAG 58 3 CGAGUA 5 CUCCGG 4 U20 1523 CDS GAGCUCACCCUGGCC 20 UAACUCGGCCAGGGUGA 584 GAGUUA 6 GCUCCG 5PAT059857-PCT-SEC01U20 1526 CDS CUCACCCUGGCCGAG 20 UCUCAACUCGGCCAGGG 58 5 UUGAGA 7 UGAGCU 6 U20 1533 CDS UGGCCGAGUUGAGGC 20 UUCUCUGCCUCAACUCG 58 6 AGAGAC 8 GCCAGG 7 U20 1538 CDS GAGUUGAGGCAGAGA 20 UAUCAGUCUCUGCCUCA 58 7 CUGAUA 9 ACUCGG 8 U20 1546 CDS GCAGAGACUGAUCCA 21 UAGAAGUGGAUCAGUC 58 8 CUUCUA 0 UCUGCCU 9 U20 1547 CDS CAGAGACUGAUCCAC 21 AGAGAAGUGGAUCAGU 59 9 UUCUCU 1 CUCUGCC 0 U21 1547 CDS CAGAGACUGAUCCAC 21 UGAGAAGUGGAUCAGU 59 0 UUCUCA 2 CUCUGCC 1 U21 1548 CDS AGAGACUGAUCCACU 21 UAGAGAAGUGGAUCAG 59 1 UCUCUG 3 UCUCUGC 2 U21 1562 CDS UUCUCUGCCAAAGAU 21 UAUGACAUCUUUGGCAG 59 2 GUCAUC 4 AGAAGU 3 U21 1571 CDS AAAGAUGUCAUCAAU 21 UGCCUCAUUGAUGACAU 59 3 GAGGCC 5 CUUUGG 4 U21 1577 CDS GUCAUCAAUGAGGCC 21 UAACCAGGCCUCAUUGA 59 4 UGGUUA 6 UGACAU 5 U21 1578 CDS UCAUCAAUGAGGCCU 21 UGAACCAGGCCUCAUUG 59 5 GGUUCC 7 AUGACA 6 U21 1598 CDS CCUGAGGACCAGCGG 21 UAGUACCCGCUGGUCCU 59 6 GUACUA 8 CAGGGA 7 U21 1600 CDS UGAGGACCAGCGGGU 21 UUCAGUACCCGCUGGUC 59 7 ACUGAC 9 CUCAGG 8 U21 1601 CDS GAGGACCAGCGGGUA 22 UGUCAGUACCCGCUGGU 59 8 CUGACA 0 CCUCAG 9 U21 1603 CDS GGACCAGCGGGUACU 22 UGGGUCAGUACCCGCUG 60 9 GACCCA 1 GUCCUC 0 U22 1695 CDS CAGCACACUCGGGGC 22 UUGUAGGCCCCGAGUGU 60 0 CUACAA 2 GCUGAC 1 U22 1696 CDS AGCACACUCGGGGCC 22 UGUGUAGGCCCCGAGUG 60 1 UACACG 3 UGCUGA 2 U22 1697 CDS GCACACUCGGGGCCU 22 UCGUGUAGGCCCCGAGU 60 2 ACACGA 4 GUGCUG 3 U22 1698 CDS CACACUCGGGGCCUA 22 UCCGUGUAGGCCCCGAG 60 3 CACGGA 5 UGUGCU 4 U22 1699 CDS ACACUCGGGGCCUAC 22 AUCCGUGUAGGCCCCGA 60 4 ACGGAU 6 GUGUGC 5 U22 1699 CDS ACACUCGGGGCCUAC 22 UUCCGUGUAGGCCCCGA 60 5 ACGGAA 7 GUGUGC 6 U22 1700 CDS CACUCGGGGCCUACA 22 UAUCCGUGUAGGCCCCG 606 CGGAUA 8 AGUGUG 7PAT059857-PCT-SEC01U22 1703 CDS UCGGGGCCUACACGG 22 UGCCAUCCGUGUAGGCC 60 7 AUGGCC 9 CCGAGU 8 U22 1708 CDS GCCUACACGGAUGGC 23 UCUGUGGCCAUCCGUGU 60 8 CACAGA 0 AGGCCC 9 U22 1709 CDS CCUACACGGAUGGCC 23 UGCUGUGGCCAUCCGUG 61 9 ACAGCA 1 UAGGCC 0 U23 1756 CDS GCUGCUGAGCUGCUC 23 AAACUGGAGCAGCUCAG 61 0 CAGUUU 2 CAGCUC 1 U23 1756 CDS GCUGCUGAGCUGCUC 23 UAACUGGAGCAGCUCAG 61 1 CAGUUA 3 CAGCUC 2 U23 1759 CDS GCUGAGCUGCUCCAG 23 UAGAAACUGGAGCAGCU 61 2 UUUCUA 4 CAGCAG 3 U23 1800 CDS GCGAGCGCAUGGAGG 23 UUUGGGCCUCCAUGCGC 61 3 CCCAAA 5 UCGCCC 4 U23 1823 CDS GGCAAGCUGGUCUGC 23 UGCCCGGCAGACCAGCU 61 4 CGGGCA 6 UGCCCC 5 U23 1829 CDS CUGGUCUGCCGGGCC 23 UUUGUGGGCCCGGCAGA 61 5 CACAAA 7 CCAGCU 6 U23 1831 CDS GGUCUGCCGGGCCCA 23 UCGUUGUGGGCCCGGCA 61 6 CAACGA 8 GACCAG 7 U23 1832 CDS GUCUGCCGGGCCCAC 23 AGCGUUGUGGGCCCGGC 61 7 AACGCU 9 AGACCA 8 U23 1832 CDS GUCUGCCGGGCCCAC 24 UGCGUUGUGGGCCCGGC 61 8 AACGCA 0 AGACCA 9 U23 1833 CDS UCUGCCGGGCCCACA 24 AAGCGUUGUGGGCCCGG 62 9 ACGCUU 1 CAGACC 0 U24 1833 CDS UCUGCCGGGCCCACA 24 UAGCGUUGUGGGCCCGG 62 0 ACGCUA 2 CAGACC 1 U24 1835 CDS UGCCGGGCCCACAAC 24 AAAAGCGUUGUGGGCCC 62 1 GCUUUU 3 GGCAGA 2 U24 1835 CDS UGCCGGGCCCACAAC 24 UAAAGCGUUGUGGGCCC 62 2 GCUUUA 4 GGCAGA 3 U24 1836 CDS GCCGGGCCCACAACGC 24 UAAAAGCGUUGUGGGCC 62 3 UUUUA 5 CGGCAG 4 U24 1860 CDS GUGAGGGUGUCUACG 24 UAAUGGCGUAGACACCC 62 4 CCAUUA 6 UCACCC 5 U24 1861 CDS UGAGGGUGUCUACGC 24 UCAAUGGCGUAGACACC 62 5 CAUUGC 7 CUCACC 6 U24 1862 CDS GAGGGUGUCUACGCC 24 UGCAAUGGCGUAGACAC 62 6 AUUGCA 8 CCUCAC 7 U24 1863 CDS AGGGUGUCUACGCCA 24 UGGCAAUGGCGUAGACA 62 7 UUGCCA 9 CCCUCA 8 U24 1867 CDS UGUCUACGCCAUUGC 25 UACCUGGCAAUGGCGUA 628 CAGGUG 0 GACACC 9PAT059857-PCT-SEC01U24 1914 CDS GCGUCCACACAGCUCC 25 UUGGUGGAGCUGUGUG 63 9 ACCAA 1 GACGCUG 0 U25 1938 CDS AGGCCAGCAUGGGGA 25 UACGGGUCCCCAUGCUG 63 0 CCCGUG 2 GCCUCA 1 U25 1943 CDS AGCAUGGGGACCCGU 25 UUGGACACGGGUCCCCA 63 1 GUCCAC 3 UGCUGG 2 U25 1945 CDS CAUGGGGACCCGUGU 25 UAGUGGACACGGGUCCC 63 2 CCACUA 4 CAUGCU 3 U25 1946 CDS AUGGGGACCCGUGUC 25 UCAGUGGACACGGGUCC 63 3 CACUGC 5 CCAUGC 4 U25 1977 CDS GCCACGUCCUCACAG 25 UGCAGCCUGUGAGGACG 63 4 GCUGCA 6 UGGCCC 5 U25 2006 CDS UGGGAGGUGGAGGAC 25 UCCAAGGUCCUCCACCU 63 5 CUUGGC 7 CCCAGU 6 U25 2017 CDS GGACCUUGGCACCCA 25 UGCUUGUGGGUGCCAAG 63 6 CAAGCA 8 GUCCUC 7 U25 2018 CDS GACCUUGGCACCCAC 25 UGGCUUGUGGGUGCCAA 63 7 AAGCCA 9 GGUCCU 8 U25 2020 CDS CCUUGGCACCCACAA 26 UGCGGCUUGUGGGUGCC 63 8 GCCGCA 0 AAGGUC 9 U25 2022 CDS UUGGCACCCACAAGC 26 UAGGCGGCUUGUGGGU 64 9 CGCCUG 1 GCCAAGG 0 U26 2025 CDS GCACCCACAAGCCGCC 26 UCACAGGCGGCUUGUGG 64 0 UGUGA 2 GUGCCA 1 U26 2027 CDS ACCCACAAGCCGCCUG 26 UAGCACAGGCGGCUUGU 64 1 UGCUG 3 GGGUGC 2 U26 2038 CDS GCCUGUGCUGAGGCC 26 UCUCGUGGCCUCAGCAC 64 2 ACGAGA 4 AGGCGG 3 U26 2039 CDS CCUGUGCUGAGGCCA 26 ACCUCGUGGCCUCAGCA 64 3 CGAGGU 5 CAGGCG 4 U26 2039 CDS CCUGUGCUGAGGCCA 26 UCCUCGUGGCCUCAGCA 64 4 CGAGGA 6 CAGGCG 5 U26 2040 CDS CUGUGCUGAGGCCAC 26 UACCUCGUGGCCUCAGC 64 5 GAGGUA 7 ACAGGC 6 U26 2044 CDS GCUGAGGCCACGAGG 26 UGCUGACCUCGUGGCCU 64 6 UCAGCA 8 CAGCAC 7 U26 2045 CDS CUGAGGCCACGAGGU 26 UGGCUGACCUCGUGGCC 64 7 CAGCCA 9 UCAGCA 8 U26 2048 CDS AGGCCACGAGGUCAG 27 UUUGGGCUGACCUCGUG 64 8 CCCAAC 0 GCCUCA 9 U26 2053 CDS ACGAGGUCAGCCCAA 27 UACUGGUUGGGCUGACC 65 9 CCAGUG 1 UCGUGG 0 U27 2054 CDS CGAGGUCAGCCCAAC 27 UCACUGGUUGGGCUGAC 650 CAGUGA 2 CUCGUG 1PAT059857-PCT-SEC01U27 2086 CDS GGAGGCCAGCAUCCA 27 UAAGCGUGGAUGCUGGC 65 1 CGCUUA 3 CUCCCU 2 U27 2120 CDS CCAGGUCUGGAAUGC 27 UACUUUGCAUUCCAGAC 65 2 AAAGUA 4 CUGGGG 3 U27 2135 CDS AAAGUCAAGGAGCAU 27 UAUUCCAUGCUCCUUGA 65 3 GGAAUC 5 CUUUGC 4 U27 2136 CDS AAGUCAAGGAGCAUG 27 UGAUUCCAUGCUCCUUG 65 4 GAAUCC 6 ACUUUG 5 U27 2140 CDS CAAGGAGCAUGGAAU 27 UCCGGGAUUCCAUGCUC 65 5 CCCGGA 7 CUUGAC 6 U27 2248 CDS GGCCUACGCCGUAGA 27 UUGUUGUCUACGGCGUA 65 6 CAACAA 8 GGCCCC 7 U27 2249 CDS GCCUACGCCGUAGAC 27 UGUGUUGUCUACGGCGU 65 7 AACACA 9 AGGCCC 8 U27 2250 CDS CCUACGCCGUAGACA 28 ACGUGUUGUCUACGGCG 65 8 ACACGU 0 UAGGCC 9 U27 2250 CDS CCUACGCCGUAGACA 28 UCGUGUUGUCUACGGCG 66 9 ACACGA 1 UAGGCC 0 U28 2251 CDS CUACGCCGUAGACAA 28 UACGUGUUGUCUACGGC 66 0 CACGUA 2 GUAGGC 1 U28 2253 CDS ACGCCGUAGACAACA 28 UACACGUGUUGUCUACG 66 1 CGUGUG 3 GCGUAG 2 U28 2254 CDS CGCCGUAGACAACAC 28 ACACACGUGUUGUCUAC 66 2 GUGUGU 4 GGCGUA 3 U28 2254 CDS CGCCGUAGACAACAC 28 UCACACGUGUUGUCUAC 66 3 GUGUGA 5 GGCGUA 4 U28 2255 CDS GCCGUAGACAACACG 28 UACACACGUGUUGUCUA 66 4 UGUGUA 6 CGGCGU 5 U28 2274 CDS UAGUCAGGAGCCGGG 28 UGACGUCCCGGCUCCUG 66 5 ACGUCA 7 ACUACA 6 U28 2277 CDS UCAGGAGCCGGGACG 28 UGCUGACGUCCCGGCUC 66 6 UCAGCA 8 CUGACU 7 U28 2293 CDS CAGCACUACAGGCAG 28 UUGGUGCUGCCUGUAGU 66 7 CACCAA 9 GCUGAC 8 U28 2296 CDS CACUACAGGCAGCAC 29 UCGCUGGUGCUGCCUGU 66 8 CAGCGA 0 AGUGCU 9 U28 2322 CDS CCGUGACAGCCGUUG 29 AGAUGGCAACGGCUGUC 67 9 CCAUCU 1 ACGGCC 0 U29 2322 CDS CCGUGACAGCCGUUG 29 UGAUGGCAACGGCUGUC 67 0 CCAUCA 2 ACGGCC 1 U29 2326 CDS GACAGCCGUUGCCAU 29 UAGCAGAUGGCAACGGC 67 1 CUGCUA 3 UGUCAC 2 U29 2330 CDS GCCGUUGCCAUCUGC 29 UCGGCAGCAGAUGGCAA 672 UGCCGA 4 CGGCUG 3PAT059857-PCT-SEC01U29 2428 3' GGCUGGGGCUGAGCU 29 UUUUAAAGCUCAGCCCC 67 3 UTR UUAAAA 5 AGCCCU 4 U29 2429 3' GCUGGGGCUGAGCUU 29 AUUUUAAAGCUCAGCCC 67 4 UTR UAAAAU 6 CAGCCC 5 U29 2429 3' GCUGGGGCUGAGCUU 29 UUUUUAAAGCUCAGCCC 67 5 UTR UAAAAA 7 CAGCCC 6 U29 2430 3' CUGGGGCUGAGCUUU 29 UAUUUUAAAGCUCAGCC 67 6 UTR AAAAUA 8 CCAGCC 7 U29 2431 3' UGGGGCUGAGCUUUA 29 UCAUUUUAAAGCUCAGC 67 7 UTR AAAUGG 9 CCCAGC 8 U29 2432 3' GGGGCUGAGCUUUAA 30 ACCAUUUUAAAGCUCAG 67 8 UTR AAUGGU 0 CCCCAG 9 U29 2432 3' GGGGCUGAGCUUUAA 30 UCCAUUUUAAAGCUCAG 68 9 UTR AAUGGA 1 CCCCAG 0 U30 2435 3' GCUGAGCUUUAAAAU 30 UGAACCAUUUUAAAGCU 68 0 UTR GGUUCA 2 CAGCCC 1 U30 2618 3' UCCCUCACUGUGGGG 30 UAAAUGCCCCACAGUGA 68 1 UTR CAUUUC 3 GGGAGG 2 U30 2619 3' CCCUCACUGUGGGGC 30 UGAAAUGCCCCACAGUG 68 2 UTR AUUUCA 4 AGGGAG 3 U30 2621 3' CUCACUGUGGGGCAU 30 UGUGAAAUGCCCCACAG 68 3 UTR UUCACA 5 UGAGGG 4 U30 2624 3' ACUGUGGGGCAUUUC 30 AAUGGUGAAAUGCCCCA 68 4 UTR ACCAUU 6 CAGUGA 5 U30 2624 3' ACUGUGGGGCAUUUC 30 UAUGGUGAAAUGCCCCA 68 5 UTR ACCAUA 7 CAGUGA 6 U30 2625 3' CUGUGGGGC AUUUCA 30 UAAUGGUGAAAUGCCCC 68 6 UTR CCAUUA 8 ACAGUG 7 U30 2643 3' UUCAAACAGGUCGAG 30 UCACAGCUCGACCUGUU 68 7 UTR CUGUGC 9 UGAAUG 8 U30 2763 3' ACCAAGGAGGCAGGA 31 UAAGAAUCCUGCCUCCU 68 8 UTR UUCUUC 0 UGGUGG 9 U30 2770 3' AGGCAGGAUUCUUCC 31 UCCAUGGGAAGAAUCCU 69 9 UTR CAUGGA 1 GCCUCC 0 U31 2837 3' GUGAGUGUGAAAGGU 31 AUCAGCACCUUUCACAC 69 0 UTR GCUGAU 2 UCACCC 1 U31 2849 3' GGUGCUGAUGGCCCU 31 UAGAUGAGGGCCAUCAG 69 1 UTR CAUCUA 3 CACCUU 2 U31 2853 3' CUGAUGGCCCUCAUC 31 UCUGGAGAUGAGGGCCA 69 2 UTR UCCAGA 4 UCAGCA 3 U31 2907 3' UAAUGGAGGCUUAGC 31 UAGAAAGCUAAGCCUCC 69 3 UTR UUUCUG 5 AUUAAU 4 U31 2916 3' CUUAGCUUUCUGGAU 31 UAUGCCAUCCAGAAAGC 694 UTR GGCAUA 6 UAAGCC 5PAT059857-PCT-SEC01U31 2917 3' UUAGCUUUCUGGAUG 31 AGAUGCCAUCCAGAAAG 69 5 UTR GCAUCU 7 CUAAGC 6 U31 2917 3' UUAGCUUUCUGGAUG 31 UGAUGCCAUCCAGAAAG 69 6 UTR GCAUCA 8 CUAAGC 7 U31 2918 3' UAGCUUUCUGGAUGG 31 UAGAUGCCAUCCAGAAA 69 7 UTR CAUCUA 9 GCUAAG 8 U31 2925 3' CUGGAUGGCAUCUAG 32 UUCUGGCUAGAUGCCAU 69 8 UTR CCAGAA 0 CCAGAA 9 U31 2930 3' UGGCAUCUAGCCAGA 32 UCAGCCUCUGGCUAGAU 70 9 UTR GGCUGG 1 GCCAUC 0 U32 3035 3' AACACCCAAAGGUGG 32 UGCAGGCCACCUUUGGG 70 0 UTR CCUGCG 2 UGUUGC 1 U32 3104 3' CUGUCUCAGCCAACCC 32 UGAGCGGGUUGGCUGA 70 1 UTR GCUCA 3 GACAGUG 2 U32 3105 3' UGUCUCAGCCAACCC 32 UGGAGCGGGUUGGCUG 70 2 UTR GCUCCA 4 AGACAGU 3 U32 3109 3' UCAGCCAACCCGCUCC 32 UUAGUGGAGCGGGUUG 70 3 UTR ACUAC 5 GCUGAGA 4 U32 3111 3' AGCCAACCCGCUCCAC 32 UGGUAGUGGAGCGGGU 70 4 UTR UACCC 6 UGGCUGA 5 U32 3190 3' GCGUGCCUGCCAAGC 32 UUGUGAGCUUGGCAGGC 70 5 UTR UCACAA 7 ACGCCC 6 U32 3248 3' CUGAAGCCAAGCCUC 32 UAAGAAGAGGCUUGGC 70 6 UTR UUCUUA 8 UUCAGAG 7 U32 3251 3' AAGCCAAGCCUCUUC 32 AAGUAAGAAGAGGCUU 70 7 UTR UUACUU 9 GGCUUCA 8 U32 3251 3' AAGCCAAGCCUCUUC 33 UAGUAAGAAGAGGCUU 70 8 UTR UUACUA 0 GGCUUCA 9 U32 3252 3' AGCCAAGCCUCUUCU 33 UAAGUAAGAAGAGGCU 71 9 UTR UACUUC 1 UGGCUUC 0 U33 3314 3' GGGAAGGGGAACACA 33 UUGGUCUGUGUUCCCCU 71 0 UTR GACCAA 2 UCCCAG 1 U33 3330 3' ACCAGGAAGCUCGGU 33 UCACUCACCGAGCUUCC 71 1 UTR GAGUGA 3 UGGUCU 2 U33 3333 3' AGGAAGCUCGGUGAG 33 UCAUCACUCACCGAGCU 71 2 UTR UGAUGG 4 UCCUGG 3 U33 3415 3' UGGCGGAGAUGCUUC 33 UCCUUAGAAGCAUCUCC 71 3 UTR UAAGGC 5 GCCAGG 4 U33 3455 3' AACAACUGUCCCUCC 33 UCUCAAGGAGGGACAGU 71 4 UTR UUGAGC 6 UGUUGG 5 U33 3458 3' AACUGUCCCUCCUUG 33 UGUGCUCAAGGAGGGAC 71 5 UTR AGCACC 7 AGUUGU 6 U33 3475 3' CACCAGCCCCACCCAA 33 UUUGCUUGGGUGGGGC 716 UTR GCAAA 8 UGGUGCU 7PAT059857-PCT-SEC01U33 3480 3' GCCCCACCCAAGCAAG 33 UUCUGCUUGCUUGGGUG 71 7 UTR CAGAA 9 GGGCUG 8 U33 3481 3' CCCCACCCAAGCAAGC 34 UGUCUGCUUGCUUGGGU 71 8 UTR AGACA 0 GGGGCU 9 U33 3485 3' ACCCAAGCAAGCAGA 34 UAAAUGUCUGCUUGCUU 72 9 UTR CAUUUA 1 GGGUGG 0 U34 3486 3' CCCAAGCAAGCAGAC 34 AUAAAUGUCUGCUUGCU 72 0 UTR AUUUAU 2 UGGGUG 1 U34 3486 3' CCCAAGCAAGCAGAC 34 UUAAAUGUCUGCUUGCU 72 1 UTR AUUUAA 3 UGGGUG 2 U34 3488 3' CAAGCAAGCAGACAU 34 AGAUAAAUGUCUGCUU 72 2 UTR UUAUCU 4 GCUUGGG 3 U34 3492 3' CAAGCAGACAUUUAU 34 UAAAAGAUAAAUGUCU 72 3 UTR CUUUUA 5 GCUUGCU 4 U34 3498 3' GACAUUUAUCUUUUG 34 UAGACCCAAAAGAUAAA 72 4 UTR GGUCUA 6 UGUCUG 5 U34 3501 3' AUUUAUCUUUUGGGU 34 UGACAGACCCAAAAGAU 72 5 UTR CUGUCC 7 AAAUGU 6 U34 3502 3' UUUAUCUUUUGGGUC 34 AGGACAGACCCAAAAGA 72 6 UTR UGUCCU 8 UAAAUG 7 U34 3502 3' UUUAUCUUUUGGGUC 34 UGGACAGACCCAAAAGA 72 7 UTR UGUCCA 9 UAAAUG 8 U34 3505 3' AUCUUUUGGGUCUGU 35 UAGAGGACAGACCCAAA 72 8 UTR CCUCUC 0 AGAUAA 9 U34 3543 3' CAACUUUUCUAGACC 35 AAAACAGGUCUAGAAA 73 9 UTR UGUUUU 1 AGUUGGC 0 U35 3543 3' CAACUUUUCUAGACC 35 UAAACAGGUCUAGAAA 73 0 UTR UGUUUA 2 AGUUGGC 1 U35 3544 3' AACUUUUCUAGACCU 35 UAAAACAGGUCUAGAA 73 1 UTR GUUUUG 3 AAGUUGG 2 U35 3547 3' UUUUCUAGACCUGUU 35 AAGCAAAACAGGUCUAG 73 2 UTR UUGCUU 4 AAAAGU 3 U35 3551 3' CUAGACCUGUUUUGC 35 ACAAAAGCAAAACAGGU 73 3 UTR UUUUGU 5 CUAGAA 4 U35 3551 3' CUAGACCUGUUUUGC 35 UCAAAAGCAAAACAGGU 73 4 UTR UUUUGA 6 CUAGAA 5 U35 3553 3' AGACCUGUUUUGCUU 35 UUACAAAAGCAAAACAG 73 5 UTR UUGUAA 7 GUCUAG 6 U35 3558 3' UGUUUUGCUUUUGUA 35 UCAAGUUACAAAAGCAA 73 6 UTR ACUUGA 8 AACAGG 7 U35 3559 3' GUUUUGCUUUUGUAA 35 UUCAAGUUACAAAAGCA 73 7 UTR CUUGAA 9 AAACAG 8 U35 3561 3' UUUGCUUUUGUAACU 36 UCUUCAAGUUACAAAAG 738 UTR UGAAGA 0 CAAAAC 9PAT059857-PCT-SEC01U35 3562 3' UUGCUUUUGUAACUU 36 AUCUUCAAGUUACAAAA 74 9 UTR GAAGAU 1 GCAAAA 0 U36 3562 3' UUGCUUUUGUAACUU 36 UUCUUCAAGUUACAAAA 74 0 UTR GAAGAA 2 GCAAAA 1 U36 3563 3' UGCUUUUGUAACUUG 36 UAUCUUCAAGUUACAAA 74 1 UTR AAGAUA 3 AGCAAA 2 U36 3564 3' GCUUUUGUAACUUGA 36 AUAUCUUCAAGUUACAA 74 2 UTR AGAUAU 4 AAGCAA 3 U36 3564 3' GCUUUUGUAACUUGA 36 UUAUCUUCAAGUUACAA 74 3 UTR AGAUAA 5 AAGCAA 4 U36 3567 3' UUUGUAACUUGAAGA 36 UAAAUAUCUUCAAGUU 74 4 UTR UAUUUA 6 ACAAAAG 5 U36 3568 3' UUGUAACUUGAAGAU 36 AUAAAUAUCUUCAAGU 74 5 UTR AUUUAU 7 UACAAAA 6 U36 3587 3' AUUCUGGGUUUUGUA 36 AAAUGCUACAAAACCCA 74 6 UTR GCAUUU 8 GAAUAA 7 U36 3587 3' AUUCUGGGUUUUGUA 36 UAAUGCUACAAAACCCA 74 7 UTR GCAUUA 9 GAAUAA 8 U36 3590 3' CUGGGUUUUGUAGCA 37 UAAAAAUGCUACAAAAC 74 8 UTR UUUUUA 0 CCAGAA 9 U36 3595 3' UUUUGUAGCAUUUUU 37 AUUAAUAAAAAUGCUA 75 9 UTR AUUAAU 1 CAAAACC 0 U37 3597 3' UUGUAGCAUUUUUAU 37 AUAUUAAUAAAAAUGC 75 0 UTR UAAUAU 2 UACAAAA 1 U37 3600 3' UAGCAUUUUUAUUAA 37 ACCAUAUUAAUAAAAA 75 1 UTR UAUGGU 3 UGCUACA 2 U37 3604 3' AUUUUUAUUAAUAUG 37 AGUCACCAUAUUAAUAA 75 2 UTR GUGACU 4 AAAUGC 3 U37 3606 3' UUUUAUUAAUAUGGU 37 AAAGUCACCAUAUUAAU 75 3 UTR GACUUU 5 AAAAAU 4 U37 3607 3' UUUAUUAAUAUGGUG 37 AAAAGUCACCAUAUUAA 75 4 UTR ACUUUU 6 UAAAAA 5 U37 3608 3' UUAUUAAUAUGGUGA 37 AAAAAGUCACCAUAUUA 75 5 UTR CUUUUU 7 AUAAAA 6 U37 3609 3' UAUUAAUAUGGUGAC 37 UAAAAAGUCACCAUAUU 75 6 UTR UUUUUA 8 AAUAAA 7 U37 3610 3' AUUAAUAUGGUGACU 37 UUAAAAAGUCACCAUAU 75 7 UTR UUUUAA 9 UAAUAA 8 U37 3612 3' UAAUAUGGUGACUUU 38 UUUUAAAAAGUCACCAU 75 8 UTR UUAAAA 0 AUUAAU 9 U37 3613 3' AAUAUGGUGACUUUU 38 AUUUUAAAAAGUCACCA 769 UTR UAAAAU 1 UAUUAA 0 * site of mRNA target is located in human PCSK9 mRNA sequence in SEQ ID NO: 2.PAT059857-PCT-SEC01

[0211] 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 modifications, for example, a modification in a nucleobase, a ribose sugar ring, and / or a phosphate group (i.e., phosphodi ester 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.

[0212] 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. In some embodiments, the dsRNA includes an antisense strand having 14 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequencePAT059857-PCT-SEC01selected from SEQ ID NOs: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. In some embodiments, the dsRNA includes an antisense strand having 21PAT059857-PCT-SEC01contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence selected from SEQ ID NOs: 382 to 760.

[0213] 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. InPAT059857-PCT-SEC01some 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760.

[0214] 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: 3 to 381. 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: 382 to 760. In some embodiments, the dsRNA includes a sense strand having 11 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotidePAT059857-PCT-SEC01sequence selected from SEQ ID NOs: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. In some embodiments, the dsRNA includes an antisense strand having 14 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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 fromPAT059857-PCT-SEC01SEQ ID NOs: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 3 to 381. 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: 382 to 760. 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: 382 to 760. 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: 382 to 760.

[0215] 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: 3 to 381 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: 382 to 760. 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: 3 to 381 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: 382 to 760. In some embodiments, the dsRNA includes (i)PAT059857-PCT-SEC01a 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: 3 to 381 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: 382 to 760. 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: 3 to 381 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: 382 to 760. 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: 3 to 381 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: 382 to 760. 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: 3 to 381 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: 382 to 760. 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: 3 to 381 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: 382 to 760.

[0216] In certain aspects, 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 PCSK9 mRNA. In certain aspects, a dsRNA as described herein may target a nucleotide region selected from regions of (i) 600-800; (ii) 800 to 1000; (iii) 1000-1200; (iv) 3100-3300; or (v) 3400-3600 of a human PCSK9 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%,PAT059857-PCT-SEC01about 95%, about 96%, about 97%, about 98%, about 99% or 100%) identity to SEQ ID NO: 2 (human PCSK9 isoform, transcript variant 1, mRNA (GenBank: NM_174936.4)). In some embodiments, the target region is selected from regions of (i) 650-750; (ii) 850-950; (iii) 1050-1150; (iv) 3200-3300; (v) 3400-3500; or (vi) 3500-3600 of a human PCSK9 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: 2 (human PCSK9 isoform, transcript variant 1, mRNA (GenBank: NM_174936.4)).

[0217] In some embodiments, the antisense strand targets a region of (i) 600-800; (ii) 800-1000; (iii) 1000-1200; (iv) 3100-3300; or (v) 3400-3600 nucleotides in a human PCSK9 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: 2 (human PCSK9 isoform, transcript variant 1, mRNA (GenBank: NM_174936.4)). In some embodiments, the antisense strand targets a region of (i) 650-750; (ii) 850-950; (iii) 1050-1150; (iv) 3200-3300; (v) 3400-3500; or (vi) 3500-3600 nucleotides in a human PCSK9 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: 2 (human PCSK9 isoform, transcript variant 1, mRNA (GenBank: NM_174936.4)). In some embodiments, the antisense strand targets a region of 3500th to 3600th nucleotides in a human PCSK9 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: 2 (human PCSK9 isoform, transcript variant 1, mRNA (GenBank: NM_174936.4)).

[0218] Exemplary selected siRNA sequences targeting the regions described herein (e.g., target region of (i) 600-800; (ii) 800 to 1000; (iii) 1000-1200; (iv) 3100-3300; or (v) 3400-3600) are shown in Table 2.PAT059857-PCT-SEC01Table 2: Selected sequences of PCSK9 siRNA (unmodified nucleotide sequences) siRN Site of Sense strand SEQ Antisense strand SEQ A No. mRN ID ID A NO. NO. Target*CUAGACCUGUUUUGCU ACAAAAGCAAAACAGGUC761 777 Pl 3551 UUUGU UAGAA CAAGCAGACAUUUAUC UAAAAGAUAAAUGUCUGC762 778 P2 3492 UUUUA UUGCU CAACUUUUCUAGACCU AAAACAGGUCUAGAAAAG763 779 P3 3543 GUUUU UUGGC GCUUUUGUAACUUGAA AUAUCUUCAAGUUACAAA764 780 P4 3564 GAUAU AGCAA UUGAAGUUGCCCCAUG UUCGACAUGGGGCAACUU765 781 P5 713 UCGAA CAAGG CCUCAUAGGCCUGGAG AUAAACUCCAGGCCUAUG766 782 P6 1102 UUUAU AGGGU AAGCCAAGCCUCUUCU AAGUAAGAAGAGGCUUGG767 783 P7 3251 UACUU CUUCA AGCCAAGCCUCUUCUU UAAGUAAGAAGAGGCUUG768 784 P8 3252 ACUUA GCUUC UUUUCUAGACCUGUUU AAGCAAAACAGGUCUAGA769 785 P9 3547 UGCUU AAAGU AGACCUGUUUUGCUUU UUACAAAAGCAAAACAGG770 786 PIO 3553 UGUAA UCUAG UUGUAACUUGAAGAUA AUAAAUAUCUUCAAGUUA771 787 Pll 3568 UUUAU CAAAA GUUUUGCUUUUGUAAC UUCAAGUUACAAAAGCAA772 788 P12 3559 UUGAA AACAG UGGUGCUAGCCUUGCG UGGAACGCAAGGCUAGCA773 789 P13 432 UUCCA CCAGC AGUGACCACCGGGAAA UUCGAUUUCCCGGUGGUC774 790 P14 881 UCGAA ACUCU UAAUGGAGGCUUAGCU UAGAAAGCUAAGCCUCCA775 791 P15 2907 UUCUA UUAAU AGACACCAGCAUACAG UCACUCUGUAUGCUGGUG776 792P16 865 AGUGA UCUAG* site of mRNA target is located in human PCSK9 mRNA sequence in SEQ ID NO: 2.

[0219] 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 fromPAT059857-PCT-SEC01SEQ ID NOs: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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:PAT059857-PCT-SEC01761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792.

[0220] 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. In some embodiments, the dsRNA includes a sense strand having 12 contiguous nucleotides differing by no more than 2PAT059857-PCT-SEC01nucleotides from the nucleotide sequence selected from SEQ ID NOs: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. In some embodiments, the dsRNA includes a sense strand having 19 contiguous nucleotides differing by no more than 2PAT059857-PCT-SEC01nucleotides from the nucleotide sequence selected from SEQ ID NOs: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792.

[0221] 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. In some embodiments, the dsRNA includes a sense strand having 14 contiguousPAT059857-PCT-SEC01nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 761 to 776. In some embodiments, the dsRNA includes an antisense strand having 14 contiguous nucleotides differing by no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. 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: 761 to 776. 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: 777 to 792. In some embodiments, the dsRNA includes a sense strand having 21 contiguous nucleotides differing byPAT059857-PCT-SEC01no more than 1 nucleotide from the nucleotide sequence selected from SEQ ID NOs: 761 to 776. 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: 777 to 792. 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: 777 to 792. 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: 777 to 792.

[0222] 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: 761 to 776 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: 777 to 792. 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: 761 to 776 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: 777 to 792. 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: 761 to 776 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: 777 to 792. 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: 761 to 776 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: 777 to 792. 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: 761 to 776 and (ii) an antisense strand forming a duplex with the sensePAT059857-PCT-SEC01strand 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: 777 to 792. 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: 761 to 776 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: 777 to 792. 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: 761 to 776 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: 777 to 792.

[0223] 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: 3 to 792), 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: 3 to 792).Modification Pattern

[0224] 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 RN A- induced silencing complex, and / or to mitigate or reduce off-targeting risk (e.g., to PCSK9 off-targeting risk).

[0225] 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 thereby imposing a longer inPAT059857-PCT-SEC01vivo 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 PCSK9). 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. In certain aspects, 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

[0226] 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. Nonlimiting 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 phosphodiester linkage), 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 non-natural nucleobase modification, a modification in a tetrahydropyran, a threofuranosyl nucleotide (TNA) modification, 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 glycol nucleic acid (GNA) modification, and a 2-O-(N-methylacetamide) modification. For example, a modified nucleotide may include a singlePAT059857-PCT-SEC01modification, or two or more modifications at the positions at which the chemical modification groups do not hinder or intervene each other.

[0227] In some embodiments, each of the modified nucleotides is independently selected from TNA, GNA, LNA, 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). In some embodiments, each of the modified nucleotides contain independently selected from TNA modification, GNA modification, LNA 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 TN As. In some embodiments, the modified nucleotides include at least one GNAs. In some embodiments, the modified nucleotides include at least one LNAs. 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).

[0228] 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), also known as “LNA havingBasea structure of e.g.,, or a pharmaceutically acceptable salt thereof, wherein is an attachment point to the adjacent nucleotides.PAT059857-PCT-SEC01

[0229] In some embodiments, a ribose ring may be replaced with a glycol moiety linked toOphosphate and the GNA includes a moiety of, or a pharmaceutically acceptable salt O BaseO.p<°I OHthereof. In some embodiments, the GNA may have a structure of or 3. pharmaceutically acceptable salt thereof. In some embodiments, the phosphodiester linkage in the GNA may be modified, e.g., with phosphorothioate group and modified GNA may have theo.p,°structureof. or a pharmaceutically acceptable salt thereof. The GNA may further include one or more substituents replacing hydrogen(s) and such modified GNA may be encompassed by the definition of GNA herein.

[0230] 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, or a pharmaceutically acceptable salt thereof. In some embodiments,the TNA may have a structureof or a pharmaceutically acceptable salt thereof. In some embodiments, the phosphodi ester linkage in the TNA may be modified, e.g., withPAT059857-PCT-SEC01phosphor othioate group and modified TNA may include a structureof “’•k. or a pharmaceutically acceptable salt thereof. The TNA may further include one or more substituents at 1', 3' and / or 4' positions and such modified TNA may be encompassed by the definition of TNA herein.

[0231] In certain aspects, the modified nucleotide 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.

[0232] 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 include one or more selected from hydrogen (i.e. deoxy), halogen (e.g., -F), substituted or unsubstituted alkyl (e.g., C1-C12 alkyl), substituted or unsubstituted heteroalkyl (e.g., -O-(Ci-Ci2 alkyl), -N-(Ci-Ci2 alkyl), -C(O)NH-(Ci-Ci2 alkyl), -NHC(O)-(Ci-Ci2 alkyl), and -C(O)-(Ci-Ci2 alkyl)). In some embodiments, the modification group may be hydrogen, -F, -O-alkyl (e.g., C1-C4 alkyl), or -O-alkoxyalkyl (e.g., -O-(Ci-C4 alkylene)-(Ci-C4 alkoxyl)). Any of the alkyl, heteroalkyl, alkylene in the disclosure are optionally substituted with one or more of hydroxyl (-OH), C1-C3 alkyl (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.

[0233] 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, thePAT059857-PCT-SEC01modified 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). In some embodiments, the modified nucleotides do not include 2'-deoxy modifications (DNA).

[0234] 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'-0-M0E or “2'-M0E”) 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'-0-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'-M0E”) 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'-0-NMA) modification.

[0235] 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 (absence of a nucleobase) or methylated nucleobase modification at nucleobase (e.g., thymine (T) or 5-methyl cytosine (5mC)).

[0236] 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 aboutPAT059857-PCT-SEC0150%, 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 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.

[0237] In certain aspects, the modified nucleotide may include a modification in a phosphate or phosphodiester linkage, in other words, an internucleoside linkage modification (e.g., phosphorothioate, phosphorodithioate, methylphosphonate, methylene phosphonate, or vinyl phosphonate (VP) linkage). In some embodiments, the linkage modification may include ■“YSH°=RTphosphorothioate (PS) having a structure of ’, which may be an Rp isomer or an Sp isomer. In some embodiments, the linkage modification may include phosphorothioate (PS)9 / SHhaving a structure of °=Rb *, which may be a stereopure Rp isomer. In some embodiments,?^SH the linkage modification may include phosphorothioate (PS) having a structure of °=Rb ' which may be a stereopure Sp isomer.PAT059857-PCT-SEC01

[0238] For example, the modified nucleotide including 3'-PS modification can be representedor a pharmaceutically acceptable salt thereof, wherein R represents H, OH or a substituent (e.g., -F, -CH3, -OMe, or MOE) and » is an attachment point to the adjacent nucleotides. In some embodiments, the 3'-PS group may be a stereopure Sp isomer. In some embodiments, the 3'-PS group may be a stereopure Rp isomer.

[0239] 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.

[0240] 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 vinyl phosphonate (VP)) attached or linked to the 5' terminal group of the first nucleotide.

[0241] In some embodiments, the first nucleotide from the 5' end of each strand (e.g., sense strand and antisense strand) includes a 5'-vinyl phosphonate (5'-VP) group that is a chemical°=pmoiety having the structure of*, or a pharmaceutically acceptable salt thereof, wherein represents the point of attachment to the 5' carbon of the pentofuranosyl sugar of a nucleotide. In some embodiments, the first nucleotide from the 5' end of each strand (e.g., sense strand andO=R antisense strand) may include (E)-vinyl phosphonate (VP) having a structure of, or a pharmaceutically acceptable salt thereof, whereinrepresents the point of attachment to the 4' carbon of the pentofuranosyl sugar. In some embodiments, the first nucleotide from the 5' end of each strand (e.g., sense strand and antisense strand) may include (Z)-vinyl phosphonate having aPAT059857-PCT-SEC01O=P. Asstructure of\= / or a pharmaceutically acceptable salt thereof, wherein5represents the point of attachment to the 4' carbon of the pentofuranosyl sugar.

[0242] In certain aspects, one or more of the modified nucleotides contain a 2' modification (e.g., 2'-0Me, 2'-F, 2'-M0E, 2'-deoxy, etc.) and an internucleoside linkage modification (e.g., phosphorothioate or (E)-vinyl phosphonate). In some embodiments, one or more of the modified nucleotides contain 2'-0Me modification and phosphorothioate group. In some embodiments, one or more of the modified nucleotides contain 2'-0Me modification and (E)-vinyl phosphonate 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)-vinyl phosphonate group. In some embodiments, one or more of the modified nucleotides contain 2'-M0E modification and phosphorothioate group. In some embodiments, one or more of the modified nucleotides contain 2'-M0E modification and (E)-vinyl phosphonate 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'-0Me modification and (E)-vinyl phosphonate group. In some embodiments, one or more of the modified nucleotides are GNA containing (E)-vinyl phosphonate group. In some embodiments, one or more of the modified nucleotides are GNA containing a phosphorothioate group. In some embodiments, one or more of the modified nucleotides are TNA containing (E)-vinyl phosphonate group. In some embodiments, one or more of the modified nucleotides are TNA containing a phosphorothioate group.

[0243] 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 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.PAT059857-PCT-SEC01Sense Strand (SS)

[0244] In certain aspects, a sense 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 sense strand is entirely made of modified nucleotides.

[0245] In certain aspects, a sense strand of the dsRNA as described herein includes two or more 2'-M0E modifications. In some embodiments, the sense strand includes two, four, six or eight 2'-M0E modifications. In some embodiments, the sense strand includes two 2'-M0E modifications. In some embodiments, the sense strand includes four 2'-M0E modifications. In some embodiments, the sense strand includes six 2'-M0E modifications. In some embodiments, the sense strand includes eight 2'-M0E modifications.

[0246] In some embodiments, the 2'-M0E modified nucleotides in the sense strand asdescribed herein include a structure of, or a pharmaceutically acceptablesalt thereof, wherein71is an attachment point to a linkage (e.g., phosphate or phosphorothioate group) or the adjacent nucleotides and “Base” is a nucleobase.

[0247] In some embodiments, the 2'-M0E modified nucleotides in the sense strand asI OH described herein include a structure of or apharmaceutically acceptable salt thereof, wherein1is an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacent nucleotides and “Base” is a nucleobase. In somePAT059857-PCT-SEC01embodiments, the 2'-M0E modified nucleotides in the sense strand as described herein include aor a pharmaceutically acceptable salt thereof.

[0248] In some embodiments, the 2'-MOE modified nucleotides include a structure ofthereof, whereinis an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacent nucleotides. In some embodiments, the 2'-M0E modified nucleotides include aor a pharmaceutically acceptable salt thereof. In some embodiments, the 2'-M0E modified nucleotides include aor a pharmaceutically acceptable salt thereof.PAT059857-PCT-SEC01

[0249] In some embodiments, the 2'-MOE modified nucleotides include a nucleotide havingacceptable salt thereof, whereinis an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacent nucleotides. In some embodiments, the 2'-M0E modified nucleotidesinclude a nucleotide having a structure of, or a pharmaceutically acceptable salt thereof. In some embodiments, the 2'-M0E modified nucleotides include anucleotide having a structure of or a pharmaceutically acceptable salt thereof. In some embodiments, the 2'-M0E modifiedPAT059857-PCT-SEC01nucleotides include a nucleotide having a structure of or a pharmaceutically acceptable salt thereof.

[0250] In some embodiments, the 2'-M0E modified nucleotides in the sense strand asNH2described herein include a nucleotide having a structure ofNH2, or a pharmaceutically acceptable salt thereof, wherein1is an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacent nucleotides. In some embodiments, the 2'-M0E modified nucleotides in the sense strand as described herein include aPAT059857-PCT-SEC01nucleotide having a structure of or a pharmaceutically acceptable salt thereof.

[0251] In some embodiments, the 2'-M0E modified nucleotides include a nucleotide havingor a pharmaceuticallyacceptable salt thereof, whereinis an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacent nucleotides. In some embodiments, the 2'-M0E modified nucleotidesinclude a nucleotide having a structure of or a pharmaceutically acceptable salt thereof.PAT059857-PCT-SEC01

[0252] In certain aspects, at least one of the 2'-M0E modified nucleotides in the sense, or a pharmaceutically acceptable salt thereof, wherein71is 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'-M0E modified nucleotides in the sense strand as describedherein has a structure of or a pharmaceutically acceptable salt thereof.

[0253] In some embodiments, the first nucleotide from the 5' end of the sense strandincludes a structure of or a pharmaceutically acceptablePAT059857-PCT-SEC01salt thereof. In some embodiments, the first nucleotide from the 5' end of the sense strandincludes a structure of, or a pharmaceutically acceptable salt thereof.

[0254] In some embodiments, the first nucleotide from the 3' end of the sense strandacceptable salt thereof. In some embodiments, the first nucleotide from the 3' end of the sensePAT059857-PCT-SEC01acceptable salt thereof, wherein1is an attachment point to a ligand. In some embodiments, thefirst nucleotide from the 3' end of the sense strand includes a structure of or a pharmaceutically acceptable salt thereof.

[0255] In certain aspects, at least one of the 2'-M0E modified nucleotides in the sensestrand as described herein has a structure of, or a pharmaceutically acceptable salt thereof, whereinis 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'-M0E modified nucleotides in the sense strand as describedherein has a structure of or a pharmaceutically acceptable salt thereof.PAT059857-PCT-SEC01

[0256] In some embodiments, the first nucleotide from the 5' end of the sense strandincludes a structure of or a pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 5' end of the sensestrand includes a structure of, or a pharmaceutically acceptable salt thereof.

[0257] In some embodiments, the first nucleotide from the 3' end of the sense strandor a pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 3' end of the sensestrand includes a structure ofPAT059857-PCT-SEC01or a pharmaceutically acceptable salt thereof, whereinis an attachment point to a ligand. In some embodiments, the first nucleotide from the 3' end of thesense strand includes a structure of or a pharmaceutically acceptable salt thereof.

[0258] In certain aspects, at least one of the 2'-M0E modified nucleotides in the sensestrand as described herein has a structureofPAT059857-PCT-SEC01or a pharmaceutically acceptable saltthereof, whereinis 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'-M0E modified nucleotides inthe sense strand as described herein has a structureof pharmaceutically acceptable salt thereof.

[0259] In some embodiments, the first nucleotide from the 5' end of the sense strandincludes a structure of or a pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 5' end of the sensePAT059857-PCT-SEC01strand includes a structure of, or a pharmaceutically acceptable salt thereof.

[0260] In some embodiments, the first nucleotide from the 3' end of the sense strand has aacceptable salt thereof. In some embodiments, the first nucleotide from the 3' end of the sensePAT059857-PCT-SEC01, or a pharmaceutically acceptable salt thereof, whereinis an attachment point to a ligand. In some embodiments, the first nucleotide from the 3' end of thesense strand has a structureof or a pharmaceutically acceptable salt thereof.

[0261] In certain aspects, at least one of the 2'-M0E modified nucleotides in the sense ostrand as described herein has a structureofacceptable salt thereof, whereinis 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'-M0E modifiedPAT059857-PCT-SEC01nucleotides in the sense strand as described herein has a structure ofor a pharmaceutically acceptable salt thereof.

[0262] In some embodiments, the first nucleotide from the 5' end of the sense strandincludes a structure of pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 5'end of the sense strand includes a structure of or a pharmaceutically acceptable salt thereof.

[0263] In some embodiments, the first nucleotide from the 3' end of the sense strand has aPAT059857-PCT-SEC01pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 3' end of the sense strand has a structure ofacceptable salt thereof, wherein71is an attachment point to a ligand. In some embodiments, the first nucleotide from the 3' end of the sense strand has a structure ofoor a pharmaceutically acceptable salt thereof.

[0264] In certain aspects, the 2'-M0E modified nucleotides locate at both 5' and 3' ends of a sense strand so as to form a structural confinement (“2'-M0E clamp”) at the sense strand termini. In some embodiments, the 2'-M0E clamps may be symmetric and having the same number of 2'-PAT059857-PCT-SEC01MOE modified nucleotides at both 5' and 3' ends of the sense strand. For example, the sense strand includes one 2'-M0E modified nucleotide at 5' end and one 2'-M0E modified nucleotide at 3' end; two 2'-M0E modified nucleotides at 5' end and two 2'-M0E modified nucleotides at 3' end; or three 2'-M0E modified nucleotides at 5' end and three 2'-M0E modified nucleotides at 3' end. In some embodiments, the 2'-M0E clamps may be asymmetric and having different numbers of 2'-M0E nucleotides at 5' and 3' ends of the sense strand. For example, the sense strand includes one 2'-M0E modified nucleotide at 5' end only; one 2'-M0E modified nucleotide at 3' end only; two 2'-M0E modified nucleotides at 5' end only; two 2'-M0E modified nucleotides at 3' end only; one 2'-M0E modified nucleotide at 5' end and two 2'-M0E modified nucleotides at 3' end; or two 2'-M0E modified nucleotides at 5' end and one 2'-M0E modified nucleotide at 3' end.

[0265] In certain aspects, the sense strand includes one 2'-M0E modified nucleotide at 5' end and one 2'-M0E modified nucleotide at 3' end. In some embodiments, the sense strand includes only one 2'-M0E modified nucleotide at 5' end and only one 2'-M0E modified nucleotide at 3' end. In some embodiments, the sense strand includes only one 2'-M0E modified nucleotide at 5' end. In some embodiments, the sense strand includes only one 2'-M0E modified nucleotide at 3' end.

[0266] In certain aspects, the sense strand includes at least two contiguous 2'-M0E modified nucleotides at 5' end and at least two 2'-M0E modified nucleotides at 3' end. In some embodiments, the sense strand includes only two 2'-M0E modified nucleotides at 5' end and only two 2'-M0E modified nucleotides at 3' end. In some embodiments, the sense strand includes only two 2'-M0E modified nucleotides at 5' end. In some embodiments, the sense strand includes only two 2'-M0E modified nucleotides at 3' end.

[0267] In certain aspects, the sense strand includes one or two contiguous 2’ -MOE modified nucleotides at 5’ end and / or one or two contiguous 2’ -MOE modified nucleotides at 3’ end, where the sense strand comprises a nucleotide sequence selected from SEQ ID NOs: 761-776. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 761. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 762. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 763. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 764. InPAT059857-PCT-SEC01some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 765. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 766. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 767. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 768. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 769. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 770. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 771. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 772. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 773. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 774. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 775. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 776.

[0268]

[0269] In certain aspects, the sense strand is 21 nucleotides in length. In some embodiments, the sense strand includes one, two, three, or four 2'-M0E 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'-M0E 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'-M0E 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'-M0E 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'-M0E modified nucleotide at the 3rd to 19th nucleotides from 5' end of the sense strands.

[0270] 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 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.

[0271] 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 twoPAT059857-PCT-SEC01TNAs at 5' end and only two TN As 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.

[0272] In some embodiments, the TNAs in the sense strand as described herein include a0structure of or a pharmaceutically acceptable salt thereof, wherein1is an attachment point to a linkage (e.g., phosphate or phosphorothioate group) or the adjacent nucleotides and “Base” is a nucleobase.

[0273] In some embodiments, the TNAs in the sense strand as described herein include astructure of or a pharmaceutically 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 embodiments, the TNAs in the dsRNA asdescribed herein include a structureof or a pharmaceutically acceptable salt thereof.PAT059857-PCT-SEC01

[0274] In some embodiments, the TNAs include a structure ofor a pharmaceutically acceptable salt thereof,wherein71is an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacentnucleotides. In some embodiments, the TNAs include a structure of pharmaceutically acceptable salt thereof.

[0275] In some embodiments, the TNAs include a nucleotide having a structure ofacceptable salt thereof, whereinis an attachment point to a terminal group (e.g., H, OH, orPAT059857-PCT-SEC01salt) or the adjacent nucleotides. In some embodiments, the TNAs include a nucleotide having aor a pharmaceutically acceptable salt thereof.

[0276] In some embodiments, the TNAs in the dsRNA as described herein include anucleotide having a structure ofNH2, or a pharmaceutically acceptable salt thereof, wherein1is an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacent nucleotides. In some embodiments, the TNAs in the dsRNA as described herein include a nucleotide having aNH2structure of or a pharmaceutically acceptable salt thereof.PAT059857-PCT-SEC01

[0277] In some embodiments, the TNAs include a nucleotide having a structure ofoo=p— OHior a pharmaceuticallyacceptable salt thereof, whereinis an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacent nucleotides. In some embodiments, the TNAs include a nucleotide having aO=p— SHstructure of < AA IAA>, or a pharmaceutically acceptable salt thereof.

[0278] In certain aspects, at least one of the TNAs in the sense strand as described hereinpharmaceutically acceptable salt thereof, wherein71is an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacent nucleotides. In some embodiments, at least one of theTNAs in the sense strand as described herein has a structure of pharmaceutically acceptable salt thereof.PAT059857-PCT-SEC01

[0279] In some embodiments, the first nucleotide from the 5' end of the sense strandincludes a structure of, or a pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 5' end of the sense strandincludes a structure of or a pharmaceutically acceptable salt thereof.

[0280] In some embodiments, the first nucleotide from the 3' end of the sense strandO=p— OH o=P— SHI Iincludes a structure of OH, OH or a pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 3' end of the sensePAT059857-PCT-SEC01or a pharmaceuticallyacceptable salt thereof and71is an attachment point to a ligand. In some embodiments, the firstnucleotide from the 3' end of the sense strand includes a structure of pharmaceutically acceptable salt thereof.

[0281] In certain aspects, at least one of the TNAs in the sense strand as described hereinpharmaceutically acceptable salt thereof, whereinis an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacent nucleotides. In some embodiments, at least one of theo O=p-— SH TNAs in the sense strand as described herein has a structure of, Q]* pharmaceutically acceptable salt thereof.PAT059857-PCT-SEC01

[0282] In some embodiments, the first nucleotide from the 5' end of the sense strandincludes a structure of or a pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 5' end of the sensestrand includes a structure of or a pharmaceutically acceptable salt thereof.

[0283] In some embodiments, the first nucleotide from the 3' end of the sense strand NHoo oO=p— OH O=p— SHI Iincludes a structure of OH OH, or a pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 3' end of the sensestrand includes a structure ofPAT059857-PCT-SEC01pharmaceutically acceptable salt thereof and1is an attachment point to a ligand. In some embodiments, the first nucleotide from the 3' end of the sense strand includes a structure ofOH, or a pharmaceutically acceptable salt thereof.

[0284] In certain aspects, at least one of the TNAs in the sense strand as described hereinpharmaceutically acceptable salt thereof, whereinis an attachment point to a terminal group (e.g., H, OH, or salt) or the adjacent nucleotides. In some embodiments, at least one of theO=P— SH TNAs in the sense strand as described herein has a structure of, or a pharmaceutically acceptable salt thereof.PAT059857-PCT-SEC01

[0285] In some embodiments, the first nucleotide from the 5' end of the sense strando oO=p-— OH o=P— - SHincludes a structure of, or a pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 5' end of the sensestrand includes a structure of or a pharmaceutically acceptable salt thereof.

[0286] In some embodiments, the first nucleotide from the 3' end of the sense strandincludes a structure of OH OH, or a pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 3' end of the sensestrand includes a structure ofPAT059857-PCT-SEC01or a pharmaceutically acceptable salt thereof and1is an attachment point to a ligand. In some embodiments, the first nucleotide from the 3' end of the sense strandincludes a structure of OH, or a pharmaceutically acceptable salt thereof.

[0287] In certain aspects, at least one of the TNAs in the sense strand as described hereino=p— SH, or a pharmaceutically acceptable salt thereof, whereinis anPAT059857-PCT-SEC01attachment point to a terminal group (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 structure of ooO=P— SHor a pharmaceutically acceptable salt thereof.

[0288] In some embodiments, the first nucleotide from the 5' end of the sense strandpharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 5'end of the sense strand includes a structure of or a pharmaceutically acceptable salt thereof.

[0289] In some embodiments, the first nucleotide from the 3' end of the sense strandincludes a structure of pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 3'PAT059857-PCT-SEC01oend of the sense strand includes a structure ofthereof and is an attachment point to a ligand. In some embodiments, the first nucleotideNH2from the 3' end of the sense strand includes a structure of OH, or a pharmaceutically acceptable salt thereof.

[0290] In certain aspects, the TN As locate at both 5' and 3' ends of a sense strand so as to form 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 TN As 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 numbersPAT059857-PCT-SEC01of TN As 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.

[0291] 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.

[0292] 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.

[0293] In certain aspects, the sense strand includes one or two contiguous 2’ -TNA modified nucleotides at 5’ end and / or one or two contiguous 2’ -TNA modified nucleotides at 3’ end, where the sense strand comprises a nucleotide sequence selected from SEQ ID NOs: 761-776. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 761. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 762. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 763. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 764. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 765. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 766. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 767. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 768. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 769. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 770. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 771. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 772. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 773. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 774. InPAT059857-PCT-SEC01some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 775. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 776.

[0294] 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. In some embodiments, the sense strand does not include a TNA at the 3rd to 19th nucleotides from 5' end of the sense strands.

[0295] 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.

[0296] 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 positionsPAT059857-PCT-SEC01from 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.

[0297] In certain aspects, a sense strand of the dsRNA as described herein includes one, two, three, or four 2'-deoxy modifications (DNA). In some embodiments, the sense strand of the dsRNA includes one 2'-deoxy modified nucleotide. In some embodiments, the sense strand includes two 2'-deoxy modified nucleotides. In some embodiments, the sense strand includes three 2'-deoxy modified nucleotides. In some embodiments, the sense strand includes four 2'-deoxy modified nucleotides.

[0298] In certain aspects, a sense strand of the dsRNA as described herein includes one, two, three, or four deoxythymidines (dT). In some embodiments, the sense strand of the dsRNA includes one deoxythymidine (dT). In some embodiments, the sense strand includes two deoxythymidines. In some embodiments, the sense strand includes three deoxythymidines (dT). In some embodiments, the sense strand includes four deoxythymidines.

[0299] In some embodiments, the sense strand is 21 nucleotides in length. In some embodiments, one or more 2'-deoxy modified nucleotides locate at 5th, 7th, 8th, and / or 9th positions from the 5' end of the sense strand. In some embodiments, one or more 2'-deoxy modified nucleotides locate at 6th, 8th, 9th, and / or 10th positions from the 5' end of the sense strand. In some embodiments, one or more 2'-deoxy modified nucleotides locate at 7th, 9th, 10th, and / or 11th positions from the 5' end of the sense strand. In some embodiments, one or more 2'-deoxy modified nucleotides locate at 8th, 10th, 11th, and / or 12th positions from the 5' end of the sense strand. In some embodiments, one or more 2'-deoxy modified nucleotides locate at 9th, 11th, 12th, and / or 13th positions from the 5' end of the sense strand.

[0300] In some embodiments, the sense strand is 21 nucleotides in length and one 2'-deoxy modified nucleotide locates at 7th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one 2'-deoxy modified nucleotide locates at 8th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one 2'-deoxy modified nucleotide locates at 9th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one 2'-deoxy modified nucleotide locates at 10th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one 2'-deoxyPAT059857-PCT-SEC01modified nucleotide locates at 11th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one 2'-deoxy modified nucleotide locates at 12th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one 2'-deoxy modified nucleotide locates at 13th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one 2'-deoxy modified nucleotide locates at 14th position from the 5' end of the sense strand.

[0301] In some embodiments, the sense strand is 21 nucleotides in length and one deoxythymidine (dT) locates at 7th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one deoxythymidine (dT) locates at 8th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one deoxythymidine (dT) locates at 9th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one deoxythymidine (dT) locates at 10th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one deoxythymidine (dT) locates at 11th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one deoxythymidine (dT) locates at 12th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one deoxythymidine (dT) locates at 13th position from the 5' end of the sense strand. In some embodiments, the sense strand is 21 nucleotides in length and one deoxythymidine (dT) locates at 14th position from the 5' end of the sense strand.

[0302] In some embodiments, when the sense strand is 21 nucleotides in length, 2'-F modified nucleotides locate at 5th, 7th, and 8th from the 5' end of the sense strand and one deoxythymidine (dT) locates at 9th position from the 5' end of the sense strand. In some embodiments, when the sense strand is 21 nucleotides in length, 2'-F modified nucleotides locate at 6th, 8th, and 9th from the 5' end of the sense strand and one deoxythymidine (dT) locates at 10th position from the 5' end of the sense strand. In some embodiments, when the sense strand is 21 nucleotides in length, 2'-F modified nucleotides locate at 7th, 9th, and 10th from the 5' end of the sense strand and one deoxythymidine (dT) locates at 11th position from the 5' end of the sense strand. In some embodiments, when the sense strand is 21 nucleotides in length, 2'-FPAT059857-PCT-SEC01modified nucleotides locate at 8th, 10th, and 11th from the 5' end of the sense strand and one deoxythymidine (dT) locates at 12th position from the 5' end of the sense strand. In some embodiments, when the sense strand is 21 nucleotides in length, 2'-F modified nucleotides locate at 9th, 11th, and 12th from the 5' end of the sense strand and one deoxythymidine (dT) locates at 13th position from the 5' end of the sense strand.

[0303] In some embodiments, the sense strand includes 2'-0Me modified nucleotides in the remaining positions in the sense strand.

[0304] In certain aspects, the sense strand includes one to six phosphorothioate (PS) linkages between nucleosides. In some embodiments, the sense strand includes one, two, three, or four phosphorothioate (PS) linkages between nucleosides.

[0305] 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.

[0306] 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 19th and 20th 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. 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.

[0307] 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 nucleotidesPAT059857-PCT-SEC01positioned 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.

[0308] 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.

[0309] 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 aPAT059857-PCT-SEC01stereopure 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.

[0310] In certain aspects, a sense strand of the dsRNA as described herein includes one or more TNAs, one or more 2'-F modified nucleotides, one or more 2'-deoxy modified nucleotides, and one or more 2'-0Me modified nucleotides. In certain aspects, a sense strand of the dsRNA as described herein consists of one or more TNAs, one or more 2'-F modified nucleotides, one or more 2'-deoxy modified nucleotides, and one or more 2'-0Me modified nucleotides.

[0311] In certain aspects, a sense strand of the dsRNA as described herein may have a Formula (I),5'-Xl-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-3' (I) wherein:each Xi, X2. X20, and X21 is independently a 2'-M0E modified nucleotide;each X3 to X19 is independently selected from a 2'-deoxy modified nucleotide, 2'-M0E modified nucleotide, 2'-F modified nucleotide, and 2'-0Me modified nucleotide.

[0312] In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence selected from the SEQ ID NOs: 761-776. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 761. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 762. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 763. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 764. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 765. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 766. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 767. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 768. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 769. In some embodiments, the nucleotide sequence of the sense strand ofPAT059857-PCT-SEC01Formula (I) comprises a sequence SEQ ID NO: 770. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 771. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 772. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 773. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 774. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 775. In some embodiments, the nucleotide sequence of the sense strand of Formula (I) comprises a sequence SEQ ID NO: 776.

[0313] In some embodiments, the first nucleotide from the 5' end of the sense strand (Xi) of Formula (I) is a 2'-M0E modified nucleotide with a nucleobase T. In some embodiments, the first nucleotide from the 5' end of the sense strand (Xi) is a 2'-M0E modified nucleotide with a nucleobase 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'-M0E modified nucleotide with a nucleobase A, 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'-M0E modified nucleotide with a nucleobase T. In some embodiments, the second nucleotide from the 5' end of the sense strand (X2) is a 2'-M0E modified nucleotide with a nucleobase G. In some embodiments, the second nucleotide from the 5' end of the sense strand (X2) is a 2'-M0E modified nucleotide with a nucleobase A. In some embodiments, the second nucleotide from the 5' end of the sense strand (X2) is a 2'-M0E modified nucleotide with a nucleobase methylated cytosine (e.g., 5-methylcytosine or N4-methylcytosine).

[0314] In some embodiments, the first nucleotide from the 3' end of the sense strand (X21) is a 2'-M0E 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'-M0E modified nucleotide with a nucleobase A. In some embodiments, the first nucleotide from the 3' end of the sense strand (X21) is a 2'-M0E modified nucleotide with a nucleobase T. In some embodiments, the second nucleotide from the 3' end of the sense strand (X20) is a 2'-M0E modified nucleotide with a nucleobase A. In some embodiments, the second nucleotide from the 3' end of the sense strand (X20) is a 2'-M0E modified nucleotide with a nucleobase G. In some embodiments, the secondPAT059857-PCT-SEC01nucleotide from the 3' end of the sense strand (X20) is a 2'-M0E modified nucleotide with a nucleobase T. In some embodiments, the second nucleotide from the 3' end of the sense strand (X20) is a 2'-M0E modified nucleotide with a nucleobase methylated cytosine (e.g., 5-methylcytosine or N4-methylcytosine).

[0315] In certain aspects, in Formula (I), X3 to X19 do not include a 2'-M0E modified nucleotide. In certain aspects, X3 to X19 do not include a TNA. In some embodiments, each X3 to X19 is selected from deoxyribonucleotide, 2'-F modified nucleotides and 2'-0Me modified nucleotides. In some embodiments, at least one of X3 to X19 is not a deoxyribonucleotide.

[0316] In some embodiments, each Xr, Xf+2, and Xr+3 is 2'-F modified nucleotide and Xr+4 is 2'-deoxy 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, and Xs are 2'-F modified nucleotides, and X9 is 2'-deoxy modified nucleotide (e.g., dT). In some embodiments, Xe, Xs, and X9 are 2'-F modified nucleotides and X10 is 2'-deoxy modified nucleotide (e.g., dT). In some embodiments, X7, X9, and Xioare 2'-F modified nucleotides, and X11 is 2'-deoxy modified nucleotide (e.g., dT). In some embodiments, Xs, X10, and X11 are 2'-F modified nucleotides, and X12 is 2'-deoxy modified nucleotide (e.g., dT). In some embodiments, X9, X11, and X12 are 2'-F modified nucleotides, and X13 is2'-deoxy modified nucleotide (e.g., dT).

[0317] In some embodiments, the sense strand includes 2'-OMe modified nucleotides in the remaining positions in the sense strand.

[0318] In some embodiments, at least two nucleotides from Xi, X2, X19, and X20 contain a 3'-PS group, respectively. In some embodiments, two nucleotides from Xi, X2, X19, and X20 contain a 3'-PS group, respectively. In some embodiments, three nucleotides from Xi, X2, X19, and X20 contain a 3'-PS group, respectively. In some embodiments, each Xi, X2, X19, and X20 contains a 3'-PS group. In some embodiments, each Xi and X2 contains a 3'-PS group.

[0319] In some embodiments, at least four from Xi, X2, X3, X4, X17, Xis, X19, and / or X20 contain 3'-PS groups. In some embodiments, four from Xi, X2, X3, X4, X17, Xis, X19, and / or X20 contain a 3'-PS group, respectively. In some embodiments, six from Xi, X2, X3, X4, X17, Xis, X19, and / or X20 contain a 3'-PS group, respectively. In some embodiments, Xi, X2, X3, X4, X17, Xis, X19, and X20 contain a 3'-PS group, respectively.PAT059857-PCT-SEC01

[0320] In some embodiments, in X3 to Xis, two to six nucleotides contain 3'-PS groups. In some embodiments, in X3 to Xis, two nucleotides contain a 3'-PS group, respectively, respectively. In some embodiments, in X3 to Xis, three nucleotides contain a 3'-PS group, respectively. In some embodiments, in X3 to Xis, four nucleotides contain a 3'-PS group, respectively. In some embodiments, in X3 to Xis, five nucleotides contain a 3'-PS group, respectively. In some embodiments, in X3 to Xis, six nucleotides contain a 3'-PS group, respectively.

[0321] In certain aspects, the sense strand includes 2'-M0E modified nucleotides positioned at the 1st, 2nd, 20th, and 21st nucleotides from the 5' end of the sense strand.

[0322] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) 2'-M0E modifications 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.

[0323] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) 2'-M0E modifications at the 1st, 2nd, 20th and / or 21st nucleotides from the 5' end of the sense strand; 2'-F modifications at 7th, 9th, and 10th nucleotides from the 5' end of the sense strand; 2'-deoxy modification at 11th nucleotide; and 2'-0Me modifications in the remaining nucleotides.

[0324] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) 2'-M0E modifications at the 1st, 2nd, 20th and / or 21st nucleotides from the 5' end of the sense strand; 2'-F modifications at 7th, 9th, and 10th nucleotides from the 5' end of the sense strand; and 2'-deoxy modification at 11th nucleotide; and(ii) 3'-PS modifications at the 1st and 2nd nucleotides from 5' end of the sense strand.

[0325] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) 2'-M0E modifications at the 1st, 2nd, 20th and / or 21st nucleotides from the 5' end of the sense strand; 2'-F modifications at 7th, 9th, and 10th nucleotides from the 5' end of the sense strand; and 2'-deoxy modification at 11th nucleotide; and(ii) 3'-PS modifications at the 1st, 2nd, 19th, and 20th nucleotides from 5' end of the sense strand.PAT059857-PCT-SEC01

[0326] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) 2'-MOE modifications at the 1st, 2nd, 20th and / or 21st nucleotides from the 5' end of the sense strand; 2'-F modifications at 7th, 9, and 10th nucleotides from the 5' end of the sense strand; 2'-deoxy modification at 11th nucleotide; 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.

[0327] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) 2'-MOE modifications at the 1st, 2nd, 20th and / or 21st nucleotides from the 5' end of the sense strand; 2'-F modifications at 7th, 9, and 10th nucleotides from the 5' end of the sense strand; 2'-deoxy modification at 11th nucleotide; 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.

[0328] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) 2'-M0E modifications at the 1st, 2nd, 20th and 21st nucleotides from the 5' end of the sense strand; 2'-F modifications at 7th, 9, and 10th nucleotides from the 5' end of the sense strand; 2'-deoxy modification at 11th nucleotide; 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.

[0329] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) 2'-M0E modifications at the 1st, 2nd, 20th and 21st nucleotides from the 5' end of the sense strand; 2'-F modifications at 7th, 9, and 10th nucleotides from the 5' end of the sense strand; 2'-deoxy modification at 11th nucleotide; 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.

[0330] In certain aspects, a sense strand of the dsRNA as described herein may have a Formula (I'),5'-Xl-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-3' (I') wherein:PAT059857-PCT-SEC01each Xi, X2. X20, and X21 is independently a TNA; andeach X3 to X19 is independently selected from 2'-deoxy modified nucleotide, 2'-MOE modified nucleotide, 2'-F modified nucleotide, and 2'-OMe modified nucleotide.

[0331] In some embodiments, the nucleotide sequence of the sense strand of Formula (F) comprises a sequence selected from the SEQ ID NOs: 761-776 of Table 2. In some embodiments, the nucleotide sequence of the sense strand of Formula (F) comprises a sequence SEQ ID NO: 761. In some embodiments, the nucleotide sequence of the sense strand of Formula (F) comprises a sequence SEQ ID NO: 762. In some embodiments, the nucleotide sequence of the sense strand of Formula (F) comprises a sequence SEQ ID NO: 763. In some embodiments, the nucleotide sequence of the sense strand of Formula (F) comprises a sequence SEQ ID NO: 764. In some embodiments, the nucleotide sequence of the sense strand of Formula (F) comprises a sequence SEQ ID NO: 765. In some embodiments, the nucleotide sequence of the sense strand of Formula (I’) comprises a sequence SEQ ID NO: 766. In some embodiments, the nucleotide sequence of the sense strand of Formula (I’) comprises a sequence SEQ ID NO: 767. In some embodiments, the nucleotide sequence of the sense strand of Formula (I’) comprises a sequence SEQ ID NO: 768. In some embodiments, the nucleotide sequence of the sense strand of Formula (F) comprises a sequence SEQ ID NO: 769. In some embodiments, the nucleotide sequence of the sense strand of Formula (I’) comprises a sequence SEQ ID NO: 770. In some embodiments, the nucleotide sequence of the sense strand of Formula (F) comprises a sequence SEQ ID NO: 771. In some embodiments, the nucleotide sequence of the sense strand of Formula (I’) comprises a sequence SEQ ID NO: 772. In some embodiments, the nucleotide sequence of the sense strand of Formula (I’) comprises a sequence SEQ ID NO: 773. In some embodiments, the nucleotide sequence of the sense strand of Formula (I’) comprises a sequence SEQ ID NO: 774. In some embodiments, the nucleotide sequence of the sense strand of Formula (I’) comprises a sequence SEQ ID NO: 775. In some embodiments, the nucleotide sequence of the sense strand of Formula (I’) comprises a sequence SEQ ID NO: 776.

[0332] In certain aspects, in Formula (I'), X3 to X19 do not include a 2'-MOE modified nucleotide. In some embodiments, each X3 to X19 is selected from deoxyribonucleotide, 2'-F modified nucleotides and 2'-OMe modified nucleotides. In some embodiments, at least one of X3 to X19 is not a deoxyribonucleotide.PAT059857-PCT-SEC01

[0333] In some embodiments, each Xf, Xf+2, and Xf+3 is 2'-F modified nucleotide and Xf+4 is 2'-deoxy 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, Xs, X7, and Xs are 2'-F modified nucleotides, and X9 is 2'-deoxy modified nucleotide (e.g., dT). In some embodiments, Xe, Xs, and X9 are 2'-F modified nucleotides and X10 is 2'-deoxy modified nucleotide (e.g., dT). In some embodiments, X7, X9, and Xioare 2'-F modified nucleotides, and X11 is 2'-deoxy modified nucleotide (e.g., dT). In some embodiments, Xs, X10, and X11 are 2'-F modified nucleotides, and X12 is 2'-deoxy modified nucleotide (e.g., dT). In some embodiments, X9, X11, and X12 are 2'-F modified nucleotides, and X13 is2'-deoxy modified nucleotide (e.g., dT).

[0334] In some embodiments, the sense strand includes 2'-OMe modified nucleotides in the remaining positions in the sense strand.

[0335] In some embodiments, at least two nucleotides from Xi, X2, X19, and X20 contain a 3'-PS group, respectively. In some embodiments, two from Xi, X2, X19, and X20 contains 3'-PS group. In some embodiments, three nucleotides from Xi, X2, X19, and X20 contain a 3'-PS group, respectively. In some embodiments, each Xi, X2, X19, and X20 contains a 3'-PS group. In some embodiments, each Xi and X2 contains a 3'-PS group.

[0336] In some embodiments, at least four from Xi, X2, X3, X4, X17, Xis, X19, and / or X20 contain 3'-PS groups. In some embodiments, four from Xi, X2, X3, X4, X17, Xis, X19, and / or X20 contain a 3'-PS group, respectively. In some embodiments, six from Xi, X2, X3, X4, X17, Xis, X19, and / or X20 contain a 3'-PS group, respectively. In some embodiments, Xi, X2, X3, X4, X17, Xis, X19, and X20 contain a 3'-PS group, respectively.

[0337] In some embodiments, in X3 to Xis, two to six nucleotides contain 3'-PS groups. In some embodiments, in X3 to Xis, two nucleotides contain a 3'-PS group, respectively. In some embodiments, in X3 to Xis, three nucleotides contain a 3'-PS group, respectively. In some embodiments, in X3 to Xis, four nucleotides contain a 3'-PS group, respectively. In some embodiments, in X3 to Xis, five nucleotides contain a 3'-PS group, respectively. In some embodiments, in X3 to Xis, six nucleotides contain a 3'-PS group, respectively.

[0338] 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.PAT059857-PCT-SEC01

[0339] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) TN As 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.

[0340] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) TN As at the 1st, 2nd, 20th and / or 21st nucleotides from the 5' end of the sense strand;2'-F modifications at 7th, 9th, and 10th nucleotides from the 5' end of the sense strand; 2'-deoxy modification at 11th nucleotide; and 2'-0Me modifications in the remaining nucleotides.

[0341] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) TN As at the 1st, 2nd, 20th and / or 21st nucleotides from the 5' end of the sense strand;2'-F modifications at 7th, 9th, and 10th nucleotides from the 5' end of the sense strand; and 2'-deoxy modification at 11th nucleotide; and(ii) 3'-PS modifications at the 1st and 2nd nucleotides from 5' end of the sense strand.

[0342] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) TN As at the 1st, 2nd, 20th and / or 21st nucleotides from the 5' end of the sense strand;2'-F modifications at 7th, 9th, and 10th nucleotides from the 5' end of the sense strand; and 2'-deoxy modification at 11th nucleotide; and(ii) 3'-PS modifications at the 1st, 2nd, 19th, and 20th nucleotides from 5' end of the sense strand.

[0343] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) TN As at the 1st, 2nd, 20th and / or 21st nucleotides from the 5' end of the sense strand;2'-F modifications at 7th, 9th, and 10th nucleotides from the 5' end of the sense strand; 2'-deoxy modification at 11th nucleotide; 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.

[0344] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) TN As at the 1st, 2nd, 20th and / or 21st nucleotides from the 5' end of the sense strand;2'-F modifications at 7th, 9th, and 10th nucleotides from the 5' end of the sensePAT059857-PCT-SEC01strand; 2'-deoxy modification at 11th nucleotide; 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.

[0345] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) TN As at the 1st, 2nd, 20th and 21st nucleotides from the 5' end of the sense strand;2'-F modifications at 7th, 9th, and 10th nucleotides from the 5' end of the sense strand; 2'-deoxy modification at 11th nucleotide; 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.

[0346] In some embodiments, the sense strand having 21 nucleotides in length includes:(i) TN As at the 1st, 2nd, 20th and 21st nucleotides from the 5' end of the sense strand;2'-F modifications at 7th, 9th, and 10th nucleotides from the 5' end of the sense strand; 2'-deoxy modification at 11th nucleotide; 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.

[0347] Exemplary modification patterns of sense strands are shown in Table 3.Table 321-mer SS 2'-MOE TNA 2'F 2'-deoxy 2'-OMe 3'-PS modification modified position modified modified modified linkage pattern No. nucleotide nucleotide nucleotide nucleotideposition position position position SSI 7, 9, 10, 11 1, 2, 3, 4, 1, 25, 6, 8, 12,13, 14, 15,16, 17, 18,19, 20, 21SS2 1, 2, 20, 21 7, 9, 10, 11 3, 4, 5, 6, 1, 28, 12, 13,14, 15, 16,17, 18, 19SS3 1, 2, 20, 21 7, 9, 10 11 3, 4, 5, 6, 1, 28, 12, 13,14, 15, 16,17, 18, 19PAT059857-PCT-SEC01SS4 1, 2, 20, 21 7, 9, 10, 11 3, 4, 5, 6, 1, 2, 19, 208, 12, 13,14, 15, 16,17, 18, 19SS5 1, 2, 20, 21 7, 9, 10 11 3, 4, 5, 6, 1, 2, 19, 208, 12, 13,14, 15, 16,17, 18, 19SS6 1, 2, 20, 7, 9, 10, 11 3, 4, 5, 6, 1, 221 8, 12, 13,14, 15, 16,17, 18, 19SS7 1, 2, 20, 7, 9, 10 11 3, 4, 5, 6, 1, 221 8, 12, 13,14, 15, 16,17, 18, 19SS8 1, 2, 20, 7, 9, 10, 11 3, 4, 5, 6, 1, 2, 19, 2021 8, 12, 13,14, 15, 16,17, 18, 19SS9 1, 2, 20, 7, 9, 10 11 3, 4, 5, 6, 1, 2, 19, 2021 8, 12, 13,14, 15, 16,17, 18, 19SS10 1, 2, 20, 21 None At least At least one At least 1, 2, 19, one among among the one among and / or 20 the remaining theremaining positions remainingpositions positionsSS11 None 1, 2, 20, At least At least one At least 1, 2, 19,21 one among among the one among and / or 20 the remaining theremaining positions remainingpositions positions

[0348] In some embodiments, the sense strand having 21 nucleotides in length has the modification pattern of SSI. 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 ofPAT059857-PCT-SEC01SS5. In some embodiments, the sense strand having 21 nucleotides in length has the modification pattern of SS6. In some embodiments, the sense strand having 21 nucleotides in length has the modification pattern of SS7. In some embodiments, the sense strand having 21 nucleotides in length has the modification pattern of SS8. In some embodiments, the sense strand having 21 nucleotides in length has the modification pattern of SS9. In some embodiments, the sense strand having 21 nucleotides in length has the modification pattern of SS1O. In some embodiments, the sense strand having 21 nucleotides in length has the modification pattern of SSI 1. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS 10, or SSI 1, and comprises a nucleotide sequence selected from SEQ ID NOs: 761-776. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS10, or SSI 1, and comprises SEQ ID NO: 761. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS10, or SS11, and comprises SEQ ID NO: 762. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS10, or SSI 1, and comprises SEQ ID NO: 763. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS10, or SSI 1, and comprises SEQ ID NO: 764. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS10, or SSI 1, and comprises SEQ ID NO: 765. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS10, or SSI 1, and comprises SEQ ID NO: 766. In some embodiments, the nucleotide sequence of the sense strand comprises SEQ ID NO: 767. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS10, or SS11, and comprises SEQ ID NO: 768. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS10, or SSI 1, and comprises SEQ ID NO: 769. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS10, or SSI 1, and comprises SEQ ID NO: 770. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS10, or SSI 1, and comprises SEQ ID NO: 771. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS10, or SSI 1, andPAT059857-PCT-SEC01comprises SEQ ID NO: 772. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS1O, or SS11, and comprises SEQ ID NO: 773. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS1O, or SSI 1, and comprises SEQ ID NO: 774. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS1O, or SSI 1, and comprises SEQ ID NO: 775. In some embodiments, the sense strand has a modification pattern of SSI, SS2, SS3, SS4, SS5, SS6, SS7, SS8, SS9, SS1O, or SSI 1, and comprises SEQ ID NO: 776. In some embodiments, the sense strand has a modification pattern of SSI and comprises SEQ ID No: 761. In some embodiments, the sense strand has a modification pattern of SS2 and comprises SEQ ID No: 761. In some embodiments, the sense strand has a modification pattern of SS3 and comprises SEQ ID No: 761. In some embodiments, the sense strand has a modification pattern of SS4 and comprises SEQ ID No: 761. In some embodiments, the sense strand has a modification pattern of SS5 and comprises SEQ ID No: 761. In some embodiments, the sense strand has a modification pattern of SS6 and comprises SEQ ID No: 761. In some embodiments, the sense strand has a modification pattern of SS7 and comprises SEQ ID No: 761. In some embodiments, the sense strand has a modification pattern of SS8 and comprises SEQ ID No: 761. In some embodiments, the sense strand has a modification pattern of SS9 and comprises SEQ ID No: 761. In some embodiments, the sense strand has a modification pattern of SSI 0 and comprises SEQ ID No: 761. In some embodiments, the sense strand has a modification pattern of SSI 1 and comprises SEQ ID No: 761.Antisense Strand (AS)

[0349] 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.

[0350] 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., phosphor othioate, phosphorodithioate, methylphosphonate, methylene phosphonate, or vinyl phosphonate (VP))PAT059857-PCT-SEC01attached or linked to the 5' terminal group of the first nucleotide) attached or linked to the 5' terminal group of the first nucleotide.

[0351] In certain aspects, the antisense strand includes 5'-vinyl phosphonate (5'-VP) group at the first nucleotide from the 5' end in the antisense strand. The “5'- VP” is a chemical moiety°=Phaving the structure of*, or a pharmaceutically acceptable salt thereof, where the wavy line represent the point of attachment to the 5' carbon of the pentofuranosyl sugar of a nucleotide.

[0352] In some embodiments, the first nucleotide from the 5' end in the antisense strandO=R.includes (E)-vinyl phosphonate (VP) having a structure of, or a pharmaceutically acceptable salt thereof, wherein the wavy line presents the point of attachment to the 4' carbon of the pentofuranosyl sugar of a nucleotide. In some embodiments, the first nucleotide from the 5'OHi ' OHend in the antisense strand includes (Z)-vinyl phosphonate having a structure of ' - ', or a pharmaceutically acceptable salt thereof, wherein the wavy line presents the point of attachment to the 4' carbon of the pentofuranosyl sugar of a nucleotide.

[0353] In some embodiments, the first nucleotide from the 5' end of the antisense strand hasO=RO Basea structure of, or a pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 5' end of thePAT059857-PCT-SEC01antisense strand has a structureof or a pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 5' end of the antisense strand has aor a pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 5' end of theantisense strand has a structure ofor a pharmaceutically acceptable salt thereof.

[0354] In some embodiments, the first nucleotide from the 5' end of the antisense strand hasor a pharmaceuticallyacceptable salt thereof, wherein1is an attachment point to the adjacent nucleotides. In some embodiments, the first nucleotide from the 5' end of the antisense strand has a structure ofPAT059857-PCT-SEC01or a pharmaceutically acceptable salt thereof. In some embodiments, the first nucleotide from the 5' end of the antisense strand has a structure ofor a pharmaceutically acceptable saltthereof, wherein1is an attachment point to the adjacent nucleotides. In some embodiments, the first nucleotide from the 5' end of the antisense strand has a structure ofor a pharmaceutically acceptable salt thereof.

[0355] In certain aspects, 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 somePAT059857-PCT-SEC01embodiments, 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.

[0356] 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, 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.

[0357] In certain aspects, an antisense strand of the dsRNA as described herein does not include a 2'-M0E modification. Alternatively, in certain aspects, the antisense strand includes one to four 2'-M0E modified nucleotides. In some embodiments, the antisense strand includes one 2'-M0E modified nucleotide. In some embodiments, the antisense strand includes two 2'-MOE modified nucleotides. In some embodiments, the antisense strand includes three 2'-M0E modified nucleotides. In some embodiments, the antisense strand includes four 2'-M0E modified nucleotides.

[0358] In certain aspects, the antisense strand includes at least one GNA. In some embodiments, the antisense strand includes only one GNA.

[0359] 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. In some embodiments, the antisense strand includes only one GNA at the 7th nucleotide from 5' end of the antisense strand.PAT059857-PCT-SEC01

[0360] In certain aspects, the antisense strand includes at least one TNA. In some embodiments, the antisense strand includes only one TNA.

[0361] In certain aspects, the antisense strand is 23 nucleotides in length. In some embodiments, the antisense strand includes only one TNA at the 3rd nucleotide from 5' end of the antisense strand. In some embodiments, the antisense strand includes only one TNA at the 4th nucleotide from 5' end of the antisense strand. In some embodiments, the antisense strand includes only one TNA at the 5th nucleotide from 5' end of the antisense strand. In some embodiments, the antisense strand includes only one TNA at the 6th nucleotide from 5' end of the antisense strand. In some embodiments, the antisense strand includes only one TNA at the 7th nucleotide from 5' end of the antisense strand. In some embodiments, the antisense strand includes only one TNA at the 8th nucleotide from 5' end of the antisense strand. In some embodiments, the antisense strand includes one or two TNA at the position 3, 5, 6, and / or 7 from 5’ end of the antisense strand, where the antisense strand has a nucleotide sequence of any one selected from SEQ ID NOs: 777-792. In some embodiments, the antisense strand includes one or two TNA at the position 3, 5, 6, and / or 7 from 5’ end of the antisense strand, where the antisense strand has a nucleotide sequence of SEQ ID NO:777. In some embodiments, the antisense strand includes one TNA at the position 3, 5, 6, or 7 from 5’ end of the antisense strand, where the antisense strand has a nucleotide sequence of any one selected from SEQ ID NOs: 777-792. In some embodiments, the antisense strand includes one TNA at the position 3, 5, 6, or 7 from 5’ end of the antisense strand, where the antisense strand has a nucleotide sequence of SEQ ID NO: 777. In some embodiments, the antisense strand includes two TNA at the position 3, and 5, 6, or 7 from 5’ end of the antisense strand, where the antisense strand has a nucleotide sequence of any one selected from SEQ ID NOs: 777-792. In some embodiments, the antisense strand includes two TNA at the position 3, and 5, 6, or 7 from 5’ end of the antisense strand, where the antisense strand has a nucleotide sequence of SEQ ID NO: 777.

[0362] In certain aspects, the antisense strand includes at least one 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC). In some embodiments, the antisense strand includes only one 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC).

[0363] In certain aspects, the antisense strand is 23 nucleotides in length. In some embodiments, the antisense strand includes only one 2'-deoxy modified nucleotide (e.g., dT, dA,PAT059857-PCT-SEC01dG, or dC) at the 3rd nucleotide from 5' end of the antisense strand. In some embodiments, the antisense strand includes only one 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC) at the 4th nucleotide from 5' end of the antisense strand. In some embodiments, the antisense strand includes only one 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC) at the 5th nucleotide from 5' end of the antisense strand. In some embodiments, the antisense strand includes only one 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC) at the 6th nucleotide from 5' end of the antisense strand. In some embodiments, the antisense strand includes only one 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC) at the 7th nucleotide from 5' end of the antisense strand. In some embodiments, the antisense strand includes only one 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC) at the 8th nucleotide from 5' end of the antisense strand.

[0364] 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 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.

[0365] In certain aspects, the antisense strand includes two to eight phosphorothioate (PS) linkages between nucleosides.

[0366] 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.PAT059857-PCT-SEC01

[0367] 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.

[0368] 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 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.

[0369] 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

[0370] In certain aspects, an antisense strand of dsRNA may have a Formula (II):5'-Xl’-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:PAT059857-PCT-SEC01each Xr to X23’ is independently selected from a 2'-deoxy modified nucleotide, 2'-F modified nucleotide, 2'-0Me modified nucleotide, 2'-M0E modified nucleotide, TNA, and GNA; andXr further includes a 5'-(E)-vinyl phosphonate group.

[0371] In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) is selected from SEQ ID NOs: 777-792. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 777. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 778. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 779. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 780. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 781. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 782. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 783. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 784. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 785. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 786. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 787. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 788. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 789. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 790. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 791. In some embodiments, the nucleotide sequence of the antisense strand of Formula (II) comprises SEQ ID NO: 792.

[0372] In certain aspects, in Formula (II), Xr to X23’ do not include a 2'-M0E modified nucleotide. In some embodiments, each Xr to X23’ is independently selected from 2'-F modified nucleotides and 2'-OMe modified nucleotides.PAT059857-PCT-SEC01

[0373] Alternatively, in certain aspects, in Formula (II), Xr to X23’ include one to four 2'-MOE modified nucleotides. In some embodiments, one of Xr, X9’, X10’, and X23’may be 2'-MOE modified nucleotide. In some embodiments, two of Xr, X9’, X10’, and X23’may be 2'-MOE modified nucleotides. In some embodiments, three of Xr, X9’, X10’, and X23’may be 2'-MOE modified nucleotides. In some embodiments, Xr, X9’, X10’, and X23’ may be 2'-MOE modified nucleotide.

[0374] In some embodiments, X2’ is a 2'-F modified nucleotide. In some embodiments, Xe’ is a 2'-F modified nucleotide. In some embodiments, Xi4’ is a 2'-F modified nucleotide. In some embodiments, Xi6’is a 2'-F modified nucleotide. In some embodiments, two of X2’, Xe’, Xi4’and Xi6’ are 2'-F modified nucleotides. In some embodiments, three of X2’, Xe’, Xi4’ and Xi6’ are 2'-F modified nucleotides. In some embodiments, each X2’, Xe’, Xi4’ and Xi6’ is a 2'-F modified nucleotide.

[0375] In some embodiments, Xr to X23’ may include at least one GNA. In some embodiments, Xr to X23’ may include only one GNA. In some embodiments, X3’ is a GNA. In some embodiments, X4’ is a GNA. In some embodiments, X5’ is a GNA. In some embodiments, Xe’ is a GNA. In some embodiments, X7’ is a GNA.

[0376] In some embodiments, Xr to X23’ may include at least one TNA. In some embodiments, Xr to X23’ may include only one TNA. In some embodiments, X3’ is a TNA. In some embodiments, X4’ is a TNA. In some embodiments, X5’ is a TNA. In some embodiments, Xe’ is a TNA. In some embodiments, X7’ is a TNA.

[0377] In some embodiments, Xr to X23’ may include at least one 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC). In some embodiments, Xr to X23’ may include only one 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC). In some embodiments, X3’ is a 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC). In some embodiments, X4’ is a 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC). In some embodiments, X5’ is a 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC). In some embodiments, Xe’ is a 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC). In some embodiments, X7’ is a 2'-deoxy modified nucleotide (e.g., dT, dA, dG, or dC). In some embodiments, Xr to X23’ may include only dT. In some embodiments, X3’ is dT. In some embodiments, X4’ is dT. In some embodiments, X5’ is dT. In some embodiments, Xe’ is dT. In some embodiments, X7’ is dT.PAT059857-PCT-SEC01

[0378] In some embodiments, the antisense strand includes 2'-OMe modified nucleotides in the remaining positions in the antisense strand.

[0379] In some embodiments, at least two from Xr, X2’, X21’, and X22’ contain 3'-PS groups. In some embodiments, two from Xr, X2’, X21’, and X22’ contain a 3'-PS group, respectively. In some embodiments, three from Xr, X2’, X21’, and X22’ contain a 3'-PS group. In some embodiments, each Xr, X2’, X21’, and X22’ contains a 3'-PS group.

[0380] In some embodiments, at least four from Xr, X2’, X3’, X4’, X19’, X20’, X21’, and X22’ contain 3'-PS groups. In some embodiments, four from Xr, X2’, X3’, X4’, X19’, X20’, X21’, and X22’ contain a 3'-PS group, respectively. In some embodiments, six from Xr, X2’, X3’, X4’, X19’, X20’, X21’, and X22’ contain a 3'-PS group, respectively. In some embodiments, Xr, X2’, X3’, X4’, X19’, X20’, X21’, and X22’ contain a 3'-PS group, respectively.

[0381] 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.

[0382] 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.

[0383] 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 22nd nucleotides from 5' end of the antisense strand.

[0384] 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;PAT059857-PCT-SEC01(ii) 2'-F modifications at 2nd, 6th, 14th, and / or 16th nucleotides 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 22nd nucleotides from 5' end of the antisense strand.

[0385] 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 5th nucleotide 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 22nd nucleotides from 5' end of the antisense strand.

[0386] 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 16th nucleotides from the 5' end of the antisense strand; GNA at 6th nucleotide 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 22nd nucleotides from 5' end of the antisense strand.

[0387] 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 7th nucleotide 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 22nd nucleotides from 5' end of the antisense strand.

[0388] In some embodiments, the antisense strand having 23 nucleotides in length includes:PAT059857-PCT-SEC01(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; TNA at 3rd nucleotide 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.

[0389] 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; TNA at 5th nucleotide 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 22nd nucleotides from 5' end of the antisense strand.

[0390] 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 16th nucleotides 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 22nd nucleotides from 5' end of the antisense strand.

[0391] 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; TNA at 7th nucleotide from the 5' end of the antisense strand; and 2'-OMe modifications in the remaining nucleotides; andPAT059857-PCT-SEC01(iii) 3'-PS modifications at the 1st, 2nd, 21st, and / or 22nd nucleotides from 5' end of the antisense strand.

[0392] 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; a 2'-deoxy modification at 5th nucleotide 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 22nd nucleotides from 5' end of the antisense strand.

[0393] 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 16th nucleotides 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 22nd nucleotides from 5' end of the antisense strand.

[0394] 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; a 2'-deoxy modification at 7th nucleotide 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 22nd nucleotides from 5' end of the antisense strand.

[0395] 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;PAT059857-PCT-SEC01(ii) 2'-F modifications at 2nd, 6th, 14th, and / or 16th nucleotides from the 5' end of the antisense strand; GNA at 3rd and 5th nucleotides 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 22nd nucleotides from 5' end of the antisense strand.

[0396] 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 16th nucleotides from the 5' end of the antisense strand; GNA at 3rd and 6th nucleotides from the 5' end of the antisense strand; and 2'-OMe modificati...

Claims

PAT059857-PCT-SEC01WHAT IS CLAIMED:

1. A double stranded RNAi (dsRNAi) agent comprising:(i) a sense strand comprising a nucleotide sequence selected from SEQ ID Nos. 3 to 381; and(ii) an antisense strand forming a duplex with the sense strand and comprising a nucleotide sequence selected from SEQ ID Nos. 382 to 760.

2. A double stranded RNAi (dsRNAi) agent comprising:(i) a sense strand comprising a nucleotide sequence selected from SEQ ID Nos. 761 to 776; and(ii) an antisense strand forming a duplex with the sense strand and comprising a nucleotide sequence selected from SEQ ID Nos. 777 to 792.

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

4. The dsRNAi agent of any one of claims 1 to 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 threofiiranosyl nucleotide (TNA) modification, a 2'-5'-linkage modification, a conformationally restricting modification, an abasic modification, a 2'- aminomodification, a 2'-O-allyl modification, 2'-C-alkyl modification, a 2'-O-alkoxyalkyl modification, a morpholino modification, a phosphoramidate modification, a nonnatural 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'-deoxyPAT059857-PCT-SEC01modification, 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.

6. The dsRNAi agent of any one of claims 1 through 5, wherein the dsRNAi agent comprises a 3'-phosphorothioate (PS) modification.

7. The dsRNAi agent of any one of claims 3 to 6, wherein each of the modified nucleotides independently comprises one or more modifications selected from 2'- deoxy modification, 2'-O-methyl (2'-0Me) modification, 2'-fluoro (2'-F) modification, 2'-O-methoxyethyl (2'-M0E) modification, the modification containing anon-natural nucleobase, TNA, GNA, 3'-phosphorothioate (PS) modification, and 5'- vinyl -phosphonate (5'-VP) modification.

8. The dsRNAi agent of any one of claims 1 to 7, wherein the sense strand comprises one or two 2'-MOE modifications positioned at the 1st and / or 2nd nucleotides from the 5' end.

9. The dsRNAi agent of any one of claims 1 to 8, wherein the sense strand comprises one or two 2'-MOE modifications positioned at the 1st and / or 2nd nucleotides from the 3' end.

10. The dsRNAi agent of any one of claims 1 to 7, wherein the sense strand comprises one or two TNAs positioned at the 1st and / or 2nd nucleotides from the 5' end.

11. The dsRNAi agent of any one of claims 1 to 7 and 10, wherein the sense strand comprises one or two TNAs positioned at the 1st and / or 2nd nucleotides from the 3' end.

12. The dsRNAi agent of any one of claims 1 to 11, wherein the antisense strand comprises a 5'-VP modification at the 1st nucleotide from the 5' end.PAT059857-PCT-SEC0113. The dsRNAi agent of any one of claims 1 to 12, wherein the antisense strand comprises a 5'-VP-2'-OMe modification at the 1st position from the 5' end.

14. The dsRNAi agent of any one of claims 1 to 13, wherein each of the sense strand and the antisense strand independently comprises two, three, four, five or six 2'-F modified nucleotides.

15. The dsRNAi agent of any one of claims 1 to 14, wherein the sense strand comprises one or two 3'-PS modifications at the 1st and / or 2nd nucleotides from the 5' end.

16. The dsRNAi agent of any one of claims 1 to 15, wherein the sense strand comprises one or two 3'-PS modifications at the 1st and / or 2nd nucleotides from the 3' end.

17. The dsRNAi agent of any one of claims 1 to 16, wherein the antisense strand comprises one or two 3'-PS modifications at the 1st and / or 2nd nucleotides from the 5' end, and / or one or two 3'-PS modifications at the 1st and / or 2nd nucleotides from the 3' end.

18. The dsRNAi agent of any one of claims 1 to 17, wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.

19. The dsRNAi agent of claim 18, wherein the sense strand comprises one to four 2'- MOE modifications positioned at the 1st, 2nd, 20th, and / or 21st nucleotides from the 5' end.

20. The dsRNAi agent of any one of claims 18 to 19, wherein the sense strand does not comprise a 2'-MOE modification at the 3rd to 19th positions from the 5' end.

21. The dsRNAi agent of claim 18, wherein the sense strand comprises one to four TNAs positioned at the 1st, 2nd, 20th, and / or 21st nucleotides from the 5' end.PAT059857-PCT-SEC0122. The dsRNAi agent of any one of claims 18 and 21, wherein the sense strand does not comprise a 2'-MOE modifications and TNA at the 3rd to 19th positions from the 5' end.

23. The dsRNAi agent of any one of claims 18 to 22, wherein the sense strand comprises two, three, or four 2'-F modifications positioned at the 7th, 9th, 10th, and / or 11th nucleotides from the 5' end.24.. The dsRNAi agent of any one of claims 18 to 23, wherein the sense strand comprises one or two 2'-deoxy modifications positioned at the 10th and / or 11th nucleotides from the 5' end.

25. The dsRNAi agent of claim 18 to 24, wherein the sense strand comprises (i) 2'-F modifications positioned at the 7th, 9th, and 10th nucleotides from the 5' end and (ii) a 2'-deoxy modification positioned at the 11th nucleotide from the 5' end.

26. The dsRNAi agent of claim 19 to 25, wherein the remaining nucleotides in the sense strand comprise 2'-OMe modifications.

27. The dsRNAi agent of any one of claims 18 to 26, wherein the antisense strand comprises a 5'-(E)-VP modification at the 1st nucleotide from the 5' end.

28. The dsRNAi agent of any one of claims 18 to 26, wherein the antisense strand comprises a 5'-(E)-VP-2'-OMe modification at the 1st nucleotide from the 5' end.

29. The dsRNAi agent of any one of claims 18 to 28, wherein the antisense strand comprises two, three, or four 2'-F modifications positioned at the 2nd, 6th, 14th, and / or 16th nucleotides from the 5' end.

30. The dsRNAi agent of any one of claims 18 to 29, wherein the antisense strand comprises 2'-F modifications positioned at the 2nd, 6th, 14th, and 16th nucleotides from the 5' end.PAT059857-PCT-SEC0131. The dsRNAi agent of any one of claims 18 to 29, wherein the antisense strand comprises 2'-F modifications positioned at the 2nd, 6th, 14th, and 16th nucleotides from the 5' end and a TNA positioned at the 3rd nucleotide from the 5' end.

32. The dsRNAi agent of any one of claims 18 to 29, wherein the antisense strand comprises 2'-F modifications positioned at the 2nd, 6th, 14th, and 16th nucleotides from the 5' end and a TNA, GNA or 2'-deoxy modification positioned at the 5th nucleotide from the 5' end.

33. The dsRNAi agent of any one of claims 18 to 29, wherein the antisense strand comprises 2'-F modifications positioned at the 2nd, 14th, and 16th nucleotides from the 5' end and a TNA, GNA or 2'-deoxy modification positioned at the 6th nucleotide from the 5' end.

34. The dsRNAi agent of any one of claims 18 to 29, wherein the antisense strand comprises 2'-F modifications positioned at the 2nd, 6th, 14th, and 16th nucleotides and a TNA, GNA or 2'-deoxy modification positioned at the 7th nucleotide.

35. The dsRNAi agent of any one of claims 27 to 34, wherein the remaining nucleotides in antisense strand comprise 2'-OMe modified modifications.

36. The dsRNAi agent of any one of claims 18 to 35, wherein the sense strand comprises one to eight 3'-PS group at the 1st, 2nd, 3rd, 4th, 17th, 18th, 19th and / or 20th nucleotides from the 5' end.

37. The dsRNAi agent of any one of claims 18 to 36, wherein the antisense strand comprises one to eight 3'-PS group at the 1st, 2nd, 3rd, 4th, 19th, 20th, 21st and / or 22nd nucleotides from the 5' end.

38. The dsRNAi agent of any one of claims 15, 16, 17, 36 and 37, wherein at least one of the 3'-PS groups in each sense strand and antisense strand has a stereopure Rp configuration.PAT059857-PCT-SEC0139. The dsRNAi agent of any one of claims 15, 16, 17, 36 and 37, wherein at least one of the 3'-PS groups in each sense strand and antisense strand has a stereopure Sp configuration.

40. A double stranded RNAi (dsRNAi) agent comprising:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andan antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andan antisense strand comprising SEQ ID NO: 859;(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;or(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883.

41. The dsRNAi agent of any one of claims 1 through 40, further comprising a ligand.

42. The dsRNAi agent of claim 1, wherein the ligand comprises a N-acetylgalactosamine (GalNAc) moiety.

43. The dsRNAi agent of claim 1 or 42, wherein the ligand has a structure of:wherein:each L1is independently a linker which may be same or different in each occurrence; L2is a linker;PAT059857-PCT-SEC01n is an integer from 1 to 3; andis an attachment point to the sense strand or the antisense strand, or to a conjugate linker conjugated to the sense strand or the antisense strand.

44. The dsRNAi agent of claim 43, wherein the ligand comprises the following structurewherein:each pl, p2, p3, ql, q2, rl, r2 and r3 is independently an integer from 0 to 12; each nl, n2, and n3 is independently an integer from 1 to 3; and“*” is an attachment point to L2.

45. The dsRNAi agent of claim 42, wherein the ligand has a structure of:PAT059857-PCT-SEC01NHAc (F),wherein: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, or to a conjugate linker conjugated to the sense strand or the antisense strand.

46. The dsRNAi agent of claim 45, wherein the ligand has a structure of:(F-l),PAT059857-PCT-SEC01HO OHwherein:each pl 1 and ql 1 is independently an integer from 0 to 12;each zl, z2, and z3 is independently an integer of 0 to 12; andis an attachment point to the sense strand or the antisense strand, or to a conjugate linker conjugated to the sense strand or the antisense strand.

47. The dsRNAi agent of any one of claims 1 to 46, wherein the ligand comprises thePAT059857-PCT-SEC01whereinis an attachment point to the sense strand or the antisense strand or to a conjugate linker conjugated to the sense strand or the antisense strand.

48. The dsRNAi agent of claim 47, wherein the ligand is conjugated to 3' end of the sense strand to form the following structure:PAT059857-PCT-SEC01OHOH or -SH.

49. The dsRNAi agent of claim 47, wherein the ligand is conjugated to 5' end of the sense strand to form the following structure:OHor a pharmaceutically acceptable salt thereof, wherein W is -OH or -SH.

50. The dsRNAi agent of claim 48 or 49, wherein W is -OH.PAT059857-PCT-SEC0151. A double stranded RNAi (dsRNAi) agent comprising:(a) a sense strand consisting of SEQ ID NO: 906, andan antisense strand consisting of SEQ ID NO: 963;(b) a sense strand consisting of SEQ ID NO: 907, andan antisense strand consisting of SEQ ID NO: 964;(c) a sense strand consisting of SEQ ID NO: 947, andan antisense strand consisting of SEQ ID NO: 1004;(d) a sense strand consisting of SEQ ID NO: 948, andan antisense strand consisting of SEQ ID NO: 1005;(e) a sense strand consisting of SEQ ID NO: 949, andan antisense strand consisting of SEQ ID NO: 1006;or(f) a sense strand consisting of SEQ ID NO: 950, andan antisense strand consisting of SEQ ID NO: 1007;wherein the ligand (L96) is conjugated to the 3' end of the sense strand to form the following schematic:OHor a pharmaceutically acceptable salt thereof, wherein W is -OH.PAT059857-PCT-SEC0152. The dsRNAi agent of any one of claims 40 to 51, wherein the dsRNAi agent is in a pharmaceutically acceptable salt form.

53. The dsRNAi agent of claim 52, wherein the pharmaceutically acceptable salt is a sodium salt.

54. A pharmaceutical composition comprising the dsRNAi agent of any one of claims 1 to 53 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

55. The pharmaceutical composition of claim 54, wherein the composition is in an aqueous solution form.

56. A method of inhibiting PCSK9 expression in a cell, the method comprising:(a) contacting the cell with the dsRNAi agent of any one of claims 1 to 53 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 54 to 55; and(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a PCSK9 gene, thereby inhibiting expression of the PCSK9 gene in the cell.

57. A method of lowering a level of low-density lipoprotein cholesterol (LDL-C) in a subject in need thereof, comprising administering to the subject the dsRNAi agent of any one of claims 1 to 53 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 54 to 55.

58. A method of treating lipidemia mediated by PCSK9 expression in a subject in need thereof, comprising administering to the subject the dsRNAi agent of any one of claims 1 to 53 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 54 to 55.

59. A method of treating or preventing atherosclerotic cardiovascular disease (ASCVD) in a subject in need thereof, comprising administering to the subject the dsRNAi agentPAT059857-PCT-SEC01of any one of claims 1 to 53 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 54 to 55.

60. A method of reducing or preventing cardiovascular event in a subject in need thereof, comprising administering to the subject the dsRNAi agent of any one of claims 1 to 53 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 54 to 55.

61. The method of claim 60, wherein the cardiovascular event is cardiovascular death, non-fatal myocardial infarction (MI), non-fatal ischemic stroke, urgent coronary revascularization, coronary heart disease (CHD) death, or any combination thereof.

62. A method of reducing or preventing a major limb adverse event (MALE) in a subject in need thereof, comprising administering to the subject the dsRNAi agent of any one of claims 1 to 53 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 54 to 55.

63. The method of claim 62, wherein the MALE is acute lower limb ischemia, lower limb amputation due to ischemia, urgent lower limb revascularization for ischemia, or any combination thereof.

64. A method of inhibiting PCSK9 expression in a cell, the method comprising:(i) contacting the cell with a dsRNAi agent, wherein the dsRNAi agent comprises:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andan antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andan antisense strand comprising SEQ ID NO: 859;(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;PAT059857-PCT-SEC01or(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883, (ii) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a PCSK9 gene, thereby inhibiting expression of the PCSK9 gene in the cell.

65. A method of lowering a level of low-density lipoprotein cholesterol (LDL-C) in a subject in need thereof, comprising administering to the subject a dsRNAi agent, wherein the dsRNAi agent comprises:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andan antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andan antisense strand comprising SEQ ID NO: 859;(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;or(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883.

66. A method of treating lipidemia mediated by PCSK9 expression in a subject in need thereof, comprising administering to the subject a dsRNAi agent, wherein the dsRNAi agent comprises:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andan antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andan antisense strand comprising SEQ ID NO: 859;PAT059857-PCT-SEC01(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;or(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883.

67. A method of treating or preventing atherosclerotic cardiovascular disease (ASCVD) in a subject in need thereof, comprising administering to the subject a dsRNAi agent, wherein the dsRNAi agent comprises:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andan antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andan antisense strand comprising SEQ ID NO: 859;(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;or(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883.

68. A method of reducing or preventing cardiovascular event in a subject in need thereof, comprising administering to the subject a dsRNAi agent, wherein the dsRNAi agent comprises:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andan antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andPAT059857-PCT-SEC01an antisense strand comprising SEQ ID NO: 859;(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;or(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883.

69. The method of claim 68, wherein the cardiovascular event is cardiovascular death, non-fatal myocardial infarction (MI), non-fatal ischemic stroke, urgent coronary revascularization, coronary heart disease (CHD) death, or any combination thereof.

70. A method of reducing or preventing a major limb adverse event (MALE) in a subject in need thereof, comprising administering to the subject a dsRNAi agent, wherein the dsRNAi agent comprises:(a) a sense strand comprising SEQ ID NO: 800, andan antisense strand comprising SEQ ID NO: 853;(b) a sense strand comprising SEQ ID NO: 801, andan antisense strand comprising SEQ ID NO: 854;(c) a sense strand comprising SEQ ID NO: 806, andan antisense strand comprising SEQ ID NO: 859;(d) a sense strand comprising SEQ ID NO: 811, andan antisense strand comprising SEQ ID NO: 864;(e) a sense strand comprising SEQ ID NO: 813, andan antisense strand comprising SEQ ID NO: 866;or(f) a sense strand comprising SEQ ID NO: 830, andan antisense strand comprising SEQ ID NO: 883.

71. The method of claim 70, wherein the MALE is acute lower limb ischemia, lower limb amputation due to ischemia, urgent lower limb revascularization for ischemia, or any combination thereof.PAT059857-PCT-SEC01 72. The method of any one of claims 64 to 71, wherein the dsRNAi agent furtherwhereinPAT059857-PCT-SEC01is an atachment point to the sense strand or the antisense strand or to a conjugate linker conjugated to the sense strand or the antisense strand.

73. The method of any one of claims 57 to 72, wherein the subject is a human.

74. The method of any one of claims 57 to 72, wherein the subject has or is diagnosed with hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, congestive heart disease (CHD) or atherosclerosis.

75. The method of any one of claims 57 to 74, wherein the dsRNAi agent or the pharmaceutical composition is administered to the subject subcutaneously or intravenously.

76. A kit comprising the dsRNAi agent of any one of claims 1 to 53 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 54 to 55.

77. The kit of claim 76, further comprising an applicator.

78. The kit of claim 77, wherein the applicator is a syringe.

79. The kit of claim 78, wherein the applicator is a pre-fdled syringe.

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