Double-stranded ribonucleic acid targeting pnpla3

Abstract

The present application belongs to the field of biomedicine, and relates to a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3, a pharmaceutically acceptable salt thereof, a ligand conjugate thereof, or a pharmaceutical composition.

Description

Double-stranded RNA targeting PNPLA3

[0001] Citation of relevant applications

[0002] This application claims priority and benefits to Chinese Patent Application No. 202411896271.X, filed with the State Intellectual Property Office of the People's Republic of China on December 20, 2024, and Chinese Patent Application No. 202511902724.X, filed with the State Intellectual Property Office of the People's Republic of China on December 16, 2025, the entire contents of which are hereby incorporated herein by reference. Technical Field

[0003] This application belongs to the field of biomedicine and relates to a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3, its pharmaceutically acceptable salt, its ligand conjugate, or a pharmaceutical composition. Background Technology

[0004] Advances in nucleic acid chemistry have greatly accelerated the clinical development of oligonucleotide drugs, including antisense oligonucleotides (ASO), splice-switching oligonucleotides (SSO), aptamers, and small interfering nucleotides (siRNA). Although these approaches utilize various target recognition and gene regulation mechanisms, their molecular structures highlight the importance of chemical modification in improving the properties and long-term durability of oligonucleotide drugs.

[0005] Unmodified RNAi reagents exhibit poor metabolic stability and are readily degraded in vivo (more than 50% degradation within one minute). Although significant progress has been made in overcoming the inherent metabolic problems of natural RNA through chemical modifications, such as the introduction of certain 2'-ribose and terminal backbone modifications, there remains a need in the field for novel modified RNAi reagents to obtain RNAi agents with enhanced efficacy, stability, reduced toxicity, and improved safety.

[0006] Nonalcoholic fatty liver disease (NAFLD) is a group of diseases characterized by a liver fat content exceeding 5%, excluding alcohol consumption and other specific pathogen infections. With the advancement of basic research, especially the recent discovery of human gene mutations, many new pathogenesis mechanisms and targets have been provided for the development of new drugs for NASH. NASH is highly correlated with genetic background, and its incidence varies by region and ethnicity. Genome-wide association studies (GWAS) have found that the Patatin-like phospholipase domain protein 3 (PNPLA3) gene variant (I148M) is more common in populations with a high incidence of fatty liver and is currently the gene mutation most strongly associated with NASH.

[0007] PNPLA3 is highly expressed in the liver, particularly in hepatocytes, and functions to hydrolyze retinate, mediating the rate-limiting step in triglyceride hydrolysis. Free fatty acids are central to the pathogenesis of NASH. Free fatty acids derived from triglyceride lipolysis in adipose tissue travel to the liver via the bloodstream. Triglycerides in lipid droplets release fatty acids back into the hepatocyte free fatty acid pool by regulating lipolysis. PNPLA3 participates in this lipolysis process, and single nucleotide mutants of PNPLA3 are closely associated with the development of NASH, reflecting the importance of this lipolysis regulation. When isoleucine at position 148 of PNPLA3 is mutated to methionine, it interferes with the entry of lipid substrates into its domain that catalyzes lipolysis, disrupting PNPLA3's lipolytic function. When fatty acids are disrupted through β-oxidation or triglyceride formation, they lead to the formation of lipotoxic substances, resulting in endoplasmic reticulum stress, oxidative stress, and activation of inflammasomes. These processes are associated with phenotypic changes in NASH, including hepatocellular damage, inflammation, stellate cell activation, and the progressive accumulation of excessive extracellular matrix. Studies have shown that approximately 50% of NASH patients have the I148M mutation phenotype of PNPLA3, and patients with the I148M mutation are more prone to hepatocellular fibrosis and have a higher mortality rate from liver disease. Currently, there are no marketed drugs targeting PNPLA3 for the treatment of NASH, but related antisense oligonucleotide drugs are undergoing active phase II clinical pharmacodynamic evaluations, and no serious adverse side effects have been reported, indicating that this target has a good safety profile for the treatment of NASH. In conclusion, PNPLA3 may be an effective and low-toxicity drug target for combating NASH.

[0008] Small interfering RNA (siRNA) can inhibit the expression of target genes by inhibiting or blocking the translation or transcription of target genes in a sequence-specific manner based on the RNA interference (RNAi) mechanism, thereby exerting an inhibitory effect at the mRNA level and achieving the purpose of treating diseases.

[0009] Invention Overview

[0010] On the one hand, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, and the sequence 1 and sequence 2 being at least substantially complementary;

[0011] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification;

[0012] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification.

[0013] On the other hand, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, said dsRNA comprising a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, said antisense strand being at least substantially complementary to a target gene, said sense strand comprising sequence 1.

[0014] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides with 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides with 2'-F modification.

[0015] On the other hand, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, and the sequence 1 and sequence 2 being at least substantially complementary.

[0016] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides with 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides with 2'-F modification.

[0017] On the other hand, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, said dsRNA comprising a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, said antisense strand being at least substantially complementary to a target gene, said antisense strand comprising sequence 2.

[0018] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification.

[0019] On the other hand, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, and the sequence 1 and sequence 2 being at least substantially complementary.

[0020] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification.

[0021] On the one hand, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, and the sequence 1 and sequence 2 being at least substantially complementary;

[0022] The sequence 1 has 19 to 21 nucleotides, and the 7th to 11th nucleotides (specifically, for example, the 7th, 8th, 9th, 10th or 11th nucleotides) at the 5' end of the sequence 1 contain one to three nucleotides containing a 2'-deoxy modification.

[0023] The sequence 2 has 19 to 23 nucleotides, and the 6th or 15th nucleotide from the 5' end to the last nucleotide (specifically, for example, the 6th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd or 23rd nucleotides) contains one to three nucleotides containing a 2'-deoxy modification.

[0024] On the other hand, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, said dsRNA comprising a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, said antisense strand being at least substantially complementary to a target gene, said sense strand comprising sequence 1.

[0025] The sequence 1 has 19 to 21 nucleotides, and the 7th to 11th nucleotides (specifically, for example, the 7th, 8th, 9th, 10th or 11th) at the 5' end of the sequence 1 contain one to three nucleotides containing a 2'-deoxy modification.

[0026] On the other hand, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, and the sequence 1 and sequence 2 being at least substantially complementary.

[0027] The sequence 1 has 19 to 21 nucleotides, and the 7th to 11th nucleotides (specifically, for example, the 7th, 8th, 9th, 10th or 11th) at the 5' end of the sequence 1 contain one to three nucleotides containing a 2'-deoxy modification.

[0028] On the other hand, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, said dsRNA comprising a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, said antisense strand being at least substantially complementary to a target gene, said antisense strand comprising sequence 2.

[0029] The sequence 2 has 19 to 23 nucleotides, and the 6th or 15th nucleotide from the 5' end to the last nucleotide (specifically, for example, the 6th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd or 23rd positions) of the sequence 2 contains one to three nucleotides containing a 2'-deoxy modification.

[0030] On the other hand, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, and the sequence 1 and sequence 2 being at least substantially complementary.

[0031] The sequence 2 has 19 to 23 nucleotides, and the 6th or 15th nucleotide from the 5' end to the last nucleotide (specifically, for example, the 6th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd or 23rd positions) of the sequence 2 contains one to three nucleotides containing a 2'-deoxy modification.

[0032] In some embodiments, each nucleotide of sequence 1 and sequence 2 is an optionally modified nucleotide (unless otherwise stated).

[0033] In some implementations, each nucleotide of sequence 1 and sequence 2 is a modified nucleotide.

[0034] On one hand, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, said dsRNA comprising a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, said antisense strand being at least substantially complementary to a target gene, said sense strand comprising the sequence shown in formula (I), and said antisense strand comprising the sequence shown in formula (II).

[0035] Formula (I): 5'-(Na) x N1N2N3N4N5N6N7N8N9N10N11N12N13N14N15N16N17N18N19-3',

[0036] Formula (II): 5'-N1'N2'N3'N4'N5'N6'N7'N8'N9'N10'N11'N12'N13'N14'N15'N16'N17'N18'N19'N20'N21'(Nb) y -3';

[0037] Each of Na, N1, N2, N3, N4, N6, N10, N11, N12, N13, N14, N15, N16, N17, N18, and N19 independently represents an unmodified or modified nucleotide.

[0038] The N5 represents a modified nucleotide, the modification comprising either a 2'-OMe modification or a 2'-F modification.

[0039] N7, N8, and N9 each independently represent a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification, and the modifications of N7, N8, and N9 are not all selected from 2'-F modification.

[0040] The x is selected from 0, 1, or 2;

[0041] The N1', N3', N4', N5', N7', N8', N9', N10', N11', N12', and N13' each independently represent an unmodified or modified nucleotide.

[0042] N2' and N14' each independently represent a modified nucleotide.

[0043] The N6' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification.

[0044] The N15', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification.

[0045] The N16' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification.

[0046] The value of y is selected from 0, 1, or 2.

[0047] On the other hand, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, said dsRNA comprising a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, said antisense strand being at least substantially complementary to a target gene, said sense strand comprising the sequence shown in formula (I).

[0048] Formula (I): 5'-(Na) x N1N2N3N4N5N6N7N8N9N10N11N12N13N14N15N16N17N18N19-3',

[0049] Each of Na, N1, N2, N3, N4, N6, N10, N11, N12, N13, N14, N15, N16, N17, N18, and N19 independently represents an unmodified or modified nucleotide.

[0050] The N5 represents a modified nucleotide, the modification comprising either a 2'-OMe modification or a 2'-F modification.

[0051] N7, N8, and N9 each independently represent a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification, and the modifications of N7, N8, and N9 are not all selected from 2'-F modification.

[0052] The x is selected from 0, 1 or 2.

[0053] On the other hand, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, said dsRNA comprising a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, said antisense strand being at least substantially complementary to a target gene, said antisense strand comprising the sequence shown in formula (II).

[0054] Formula (II): 5'-N1'N2'N3'N4'N5'N6'N7'N8'N9'N10'N11'N12'N13'N14'N15'N16'N17'N18'N19'N20'N21'(Nb) y -3';

[0055] Wherein, N1', N3', N4', N5', N7', N8', N9', N10', N11', N12', and N13' each independently represent an unmodified or modified nucleotide.

[0056] N2' and N14' each independently represent a modified nucleotide.

[0057] The N6' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification.

[0058] The N15', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification.

[0059] The N16' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification.

[0060] The value of y is selected from 0, 1, or 2.

[0061] In some implementations, x is selected from 0.

[0062] In some embodiments, x is selected from 1, then Na represents a modified nucleotide Na1.

[0063] In some embodiments, x is selected from 2, then Na represents two modified nucleotides Na1Na2, wherein Na1 is linked to N1.

[0064] In some implementations, y is selected from 0.

[0065] In some embodiments, y is selected from 1, then Nb represents a modified nucleotide Nb1.

[0066] In some embodiments, y is selected from 2, then Nb represents two modified nucleotides Nb1Nb2, wherein Nb1 is connected to N21'.

[0067] In another aspect, this application provides a double-stranded ribonucleic acid (dsRNA) and a pharmaceutically acceptable salt thereof, the double-stranded ribonucleic acid (dsRNA) as described in this application, and the pharmaceutically acceptable salt as described in this application.

[0068] In another aspect, this application provides a double-stranded ribonucleic acid (dsRNA) or a ligand conjugate thereof, wherein the double-stranded ribonucleic acid (dsRNA) is as described in this application, and the ligand is as described in this application.

[0069] In another aspect, this application provides a pharmaceutical composition comprising the dsRNA of this application, a pharmaceutically acceptable salt thereof or a ligand conjugate thereof; and optionally a pharmaceutically acceptable excipient (carrier or excipient).

[0070] In another aspect, this application provides a kit for treating and / or preventing diseases related to a target gene, comprising the dsRNA of this application, a pharmaceutically acceptable salt thereof, a ligand conjugate thereof, or a pharmaceutical composition thereof; and optionally including instructions for use for treating and / or preventing diseases related to the target gene.

[0071] In another aspect, this application provides a method for treating and / or preventing diseases associated with a target gene, comprising administering a therapeutically effective amount of the application's dsRNA, a pharmaceutically acceptable salt thereof, a ligand conjugate thereof, or a pharmaceutical composition to a subject (hereinafter also referred to as a subject).

[0072] On the other hand, this application provides the use of the dsRNA of this application, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition in the preparation of medicaments for treating and / or preventing diseases related to the target gene.

[0073] On the other hand, this application provides the use of the dsRNA of this application, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition in the treatment and / or prevention of diseases related to the target gene.

[0074] On the other hand, this application provides the dsRNA of this application, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition for the treatment and / or prevention of diseases related to the target gene.

[0075] In some embodiments, the dsRNA of this application, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition is used as a single therapeutic agent for treating and / or preventing diseases associated with the target gene.

[0076] In some embodiments, the dsRNA of this application, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition may be used in combination with other therapeutic agents for treating and / or preventing diseases associated with the target gene.

[0077] Invention Details

[0078] The following will describe exemplary embodiments of this application. However, those skilled in the art will understand that the scope of protection of this application is not limited thereto. Various modifications, alterations or changes can be made based on the spirit and concept of this application, and the content of such modifications, alterations or changes still falls within the scope of this application.

[0079] In some embodiments, this application also provides a dsRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, said dsRNA comprising a sense strand and an antisense strand, said sense strand and antisense strand having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the full length of the nucleotide sequences of the sense strand and antisense strand described herein.

[0080] In some embodiments, the dsRNA of this application is siRNA.

[0081] In some embodiments, the dsRNA of this application, its pharmaceutically acceptable salt, or its ligand conjugate is an RNAi drug.

[0082] Length of double chain

[0083] In some embodiments, the positive chain has 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 nucleotides, or a number of nucleotides within the range of the above values. In some embodiments, the positive chain has 19 to 35 nucleotides, 19 to 30 nucleotides, 19 to 27 nucleotides, 19 to 25 nucleotides, 19 to 23 nucleotides, or 19 to 21 nucleotides.

[0084] In some embodiments, the antisense strand has 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 nucleotides, or a number of nucleotides within the range of the above values. In some embodiments, the antisense strand has 19 to 35 nucleotides, 19 to 30 nucleotides, 19 to 27 nucleotides, 19 to 25 nucleotides, 19 to 23 nucleotides, 21 to 23 nucleotides, 21 to 25 nucleotides, 21 to 27 nucleotides, or 21 to 30 nucleotides.

[0085] In some embodiments, the positive strand has 19 to 21 nucleotides. In other embodiments, the positive strand has 19 or 21 nucleotides.

[0086] In some embodiments, the antisense strand has 21 to 23 nucleotides. In some embodiments, the dsRNA comprises a sense strand and an antisense strand, the antisense strand having 21 or 23 nucleotides.

[0087] In some embodiments, sequence 1 has 19, 20, or 21 nucleotides.

[0088] In some embodiments, sequence 2 has 19, 20, 21, 22, or 23 nucleotides.

[0089] Complementary or mismatched

[0090] In some embodiments, the antisense strand of the dsRNA of this application, its pharmaceutically acceptable salt, or its ligand conjugate has a mismatch of 1 to 5 bases with the target gene. In some embodiments, the antisense strand of the dsRNA of this application, its pharmaceutically acceptable salt, or its ligand conjugate has a mismatch of 1, 2, 3, 4, or 5 bases with the target gene.

[0091] In some embodiments, the antisense strand of the dsRNA of this application, its pharmaceutically acceptable salt, or its ligand conjugate has sufficient complementarity with the target gene to produce RNA interference (RNAi).

[0092] In some implementations, the at least substantially complementary includes substantially complementary and fully complementary.

[0093] In some embodiments, the antisense strand being at least substantially complementary to the target gene means that the antisense strand and the target gene have 3, 2, or 1 base mismatches, or that the antisense strand and the target gene are completely complementary.

[0094] In some embodiments, the site where the antisense strand has a base mismatch with the target gene is selected from the first position of the 5' end of the antisense strand, the second position of the 5' end of the antisense strand, the first position of the 3' end of the antisense strand, or the second position of the 3' end of the antisense strand.

[0095] In some implementations, the antisense strand is completely complementary to the target gene.

[0096] In some implementations, the sense strand and the antisense strand are at least substantially complementary, meaning that the sense strand and the antisense strand have 3, 2, or 1 base mismatches.

[0097] In some embodiments, the sites where the sense and antisense strands have base mismatches are selected from one or more sites (e.g., 3, 2, or 1 sites) selected from the following positions: position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8, position 9, position 10, position 11, position 12, position 13, position 14, position 15, position 16, position 17, position 18, position 19, position 20, or position 21 of the sense strand.

[0098] In some embodiments, the site where the sense strand and antisense strand have a base mismatch is selected from the 13th, 14th, 16th, 17th, or 19th position of the 5' end of the sense strand.

[0099] In some embodiments, the site where the sense strand and antisense strand have a base mismatch is selected from the first position of the 5' end of the antisense strand, the second position of the 5' end of the antisense strand, the first position of the 3' end of the antisense strand, or the second position of the 3' end of the antisense strand.

[0100] In some implementations, the justice chain and the antisense chain are completely complementary.

[0101] In some embodiments, sequence 2 is at least substantially complementary to the target gene, wherein substantially complementary means that sequence 2 has 3, 2 or 1 base mismatch with the target gene.

[0102] In some embodiments, the site where the sequence 2 has a base mismatch with the target gene is selected from the 1st position of the 5' end, the 2nd position of the 5' end, the 1st position of the 3' end, or the 2nd position of the 3' end of the sequence 2.

[0103] In some implementations, sequence 2 is completely complementary to the target gene.

[0104] In some embodiments, sequence 1 and sequence 2 are at least substantially complementary, meaning that sequence 1 and sequence 2 have 3, 2, or 1 base mismatches.

[0105] In some embodiments, the sites where the sense and antisense strands have base mismatches are selected from the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, and 2nd positions of the 5' end of the antisense strand. One or more sites (e.g., 3, 2 or 1 sites) from the following positions: 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22 or 23 of the 5' end of the antisense chain.

[0106] In some embodiments, the sites where the sequences 1 and 2 have base mismatches are selected from one or more sites (e.g., 3, 2, or 1 sites) at the 5' end of sequence 1, specifically at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21.

[0107] In some embodiments, the sites where the bases of sequence 1 and sequence 2 are mismatched are selected from one or more sites (e.g., 3, 2 or 1 sites) at the 13th, 14th, 16th, 17th or 19th position of the 5' end of sequence 1.

[0108] In some embodiments, the sites where the sequences 1 and 2 have base mismatches are selected from one or more sites (e.g., 3, 2, or 1 sites) of the 5' end of sequence 2, specifically the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, or 23rd positions.

[0109] In some embodiments, the site where the bases of sequence 1 and sequence 2 are mismatched is selected from the 1st position of the 5' end, the 2nd position of the 5' end, the 1st position of the 3' end, or the 2nd position of the 3' end of sequence 1.

[0110] In some implementations, sequence 1 and sequence 2 are completely complementary.

[0111] In some embodiments, the sequence shown in formula (II) is at least substantially complementary to the target gene, wherein substantially complementary means that the sequence shown in formula (II) has 3, 2 or 1 base mismatch with the target gene.

[0112] In some embodiments, the site where the sequence shown in Formula (II) has a base mismatch with the target gene is selected from the 1st position at the 5' end, the 2nd position at the 5' end, the 1st position at the 3' end, or the 2nd position at the 3' end of the sequence shown in Formula (II).

[0113] In some embodiments, the sequence represented by formula (II) is completely complementary to the target gene.

[0114] In some embodiments, the sequence shown in formula (I) is at least substantially complementary to the sequence shown in formula (II), wherein substantially complementary means that the sequence shown in formula (I) and the sequence shown in formula (II) have 3, 2 or 1 base mismatches.

[0115] In some embodiments, the sites where the sequence shown in Formula (I) and the sequence shown in Formula (II) have base mismatches are selected from one or more sites (e.g., 3, 2 or 1 sites) of the 5' end 1, 5' end 2, 5' end 3, 5' end 4, 5' end 5, 5' end 6, 5' end 7, 5' end 8, 5' end 9, 5' end 10, 5' end 11, 5' end 12, 5' end 13, 5' end 14, 5' end 15, 5' end 16, 5' end 17, 5' end 18 or 5' end 19 of the sequence shown in Formula (I).

[0116] In some embodiments, the sites where the sequence shown in Formula (I) and the sequence shown in Formula (II) have base mismatches are selected from one or more sites (e.g., three, two or one sites) of Na1, Na2, N1, N2, N3, N4, N5, N6, N7, N8, N9, N10, N11, N12, N13, N14, N15, N16, N17, N18 or N19.

[0117] In some embodiments, the sites where the sequence shown in Formula (I) and the sequence shown in Formula (II) have base mismatches are selected from one or more sites (e.g., 3, 2 or 1 sites) at the 13th, 14th, 16th, 17th or 19th position of the 5' end of the sequence shown in Formula (I).

[0118] In some embodiments, the sites where the sequence shown in Formula (I) and the sequence shown in Formula (II) have base mismatches are selected from one or more sites (e.g., 3, 2 or 1 sites) of N13, N14, N16, N17 or N19.

[0119] In some embodiments, the sites where the sequence shown in Formula (I) and the sequence shown in Formula (II) have base mismatches are selected from one or more sites (e.g., 3, 2, or 1 sites) of the 5' end 1, 5' end 2, 5' end 3, 5' end 4, 5' end 5, 5' end 6, 5' end 7, 5' end 8, 5' end 9, 5' end 10, 5' end 11, 5' end 12, 5' end 13, 5' end 14, 5' end 15, 5' end 16, 5' end 17, 5' end 18, 5' end 19, 5' end 20, or 5' end 21 of the sequence shown in Formula (II).

[0120] In some embodiments, the sites where the sequence shown in Formula (I) and the sequence shown in Formula (II) have base mismatches are selected from one or more sites (e.g., three, two or one sites) among N1', N2', N3', N4', N5', N6', N7', N8', N9', N10', N11', N12', N13', N14', N15', N16', N17', N18', N19', N20', N21', Nb1 or Nb2.

[0121] In some embodiments, the sites where the sequence shown in Formula (I) and the sequence shown in Formula (II) have base mismatches are selected from the 1st position at the 5' end, the 2nd position at the 5' end, the 1st position at the 3' end, or the 2nd position at the 3' end of the sequence shown in Formula (I).

[0122] In some embodiments, the sites where the sequence shown in Formula (I) and the sequence shown in Formula (II) have base mismatches are selected from one or more sites (e.g., 3, 2 or 1 sites) of N1', N2', N20' or N21'.

[0123] In some embodiments, the sequence shown in (I) is completely complementary to the sequence shown in (II).

[0124] Dual-chain region

[0125] In some embodiments, the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40, 19 to 35, 19 to 30, or 19 to 25 nucleotides. In some embodiments, the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 23 nucleotides.

[0126] In the double-stranded ribonucleic acid (dsRNA), pharmaceutically acceptable salts thereof, or ligand conjugates thereof of this application, the dsRNA comprises a double-stranded region formed by the complementary sequences of the sense and antisense strands, and the dsRNA comprises one or two blunt ends, or one or two overhanging ends.

[0127] In the double-stranded RNA (dsRNA), its pharmaceutically acceptable salt, or its ligand conjugates of this application, the dsRNA comprises a double-stranded region formed by the complementary sequences of the sense and antisense strands. The dsRNA includes one or two blunt ends, or one or two overhanging ends, located at one or both ends of the double-stranded region. Specifically, for example, the 3' end of the sense strand forms a blunt end with the 5' end of the antisense strand; the 5' end of the sense strand forms a blunt end with the 3' end of the antisense strand; the 3' end of the sense strand forms an overhanging end with the 5' end of the antisense strand; or the 5' end of the sense strand forms an overhanging end with the 3' end of the antisense strand.

[0128] In some implementations, the sequences of the sense and antisense strands are complementary to form a double-stranded region, the length of which is selected from 12 to 40 pairs of nucleotides.

[0129] In some embodiments, the sense and antisense strands complement each other to form a double-stranded region, the length of which is selected from 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40 pairs of nucleotides, or a number of nucleotides within the range formed by the above values.

[0130] In some embodiments, the sense and antisense strands complement each other to form a double-stranded region, the length of which is selected from 14 to 38 pairs of nucleotides, 15 to 36 pairs of nucleotides, 16 to 34 pairs of nucleotides, 17 to 32 pairs of nucleotides, 18 to 30 pairs of nucleotides, 18 to 28 pairs of nucleotides, 19 to 26 pairs of nucleotides, 19 to 25 pairs of nucleotides, 19 to 24 pairs of nucleotides, 19 to 23 pairs of nucleotides, or 19 to 21 pairs of nucleotides.

[0131] In some implementations, the sense and antisense strands complement each other to form a double-stranded region, the length of which is selected from 19 or 21 pairs of nucleotides.

[0132] In some embodiments, the sense chain and the antisense chain complement each other to form a double-stranded region, and the 3' end of the sense chain and the 5' end of the antisense chain complement each other to form a flat end.

[0133] In some implementations, the sense chain and the antisense chain complement each other to form a double-chain region, and the 3' end of the sense chain and the 5' end of the antisense chain complement each other to form a protruding end.

[0134] In some implementations, the sense chain and the antisense chain complement each other to form a double-chain region, and the 5' end of the sense chain and the 3' end of the antisense chain complement each other to form a protruding end.

[0135] In some embodiments, sequence 1 and sequence 2 are complementary to form a double-stranded region, the length of which is selected from 17 to 23 pairs of nucleotides.

[0136] In some embodiments, sequence 1 and sequence 2 are complementary to form a double-stranded region, the length of which is selected from 17, 18, 19, 20, 21, 22 or 23 pairs of nucleotides, or a number of nucleotides within the range of the above values.

[0137] In some embodiments, sequence 1 and sequence 2 are complementary to form a double-stranded region, the length of which is selected from 18 to 22 pairs of nucleotides or 19 to 21 pairs of nucleotides.

[0138] In some embodiments, sequence 1 and sequence 2 are complementary to form a double-stranded region, the length of which is selected from 19 or 21 pairs of nucleotides.

[0139] In some embodiments, sequence 1 and sequence 2 are complementary to form a double-stranded region, and the first nucleotide at the 3' end of sequence 1 is complementary to the first nucleotide at the 5' end of sequence 2.

[0140] In some embodiments, sequence 1 and sequence 2 are complementary to form a double-stranded region, and the 3' end of sequence 1 and the 5' end of sequence 2 are complementary to form a flat end.

[0141] In some embodiments, sequence 1 and sequence 2 are complementary to form a double-stranded region, and the 5' end of sequence 1 and the 3' end of sequence 2 are complementary to form a flat end.

[0142] In some embodiments, sequence 1 and sequence 2 are complementary to form a double-stranded region, and the 3' end of sequence 1 and the 5' end of sequence 2 are complementary to form a protruding end.

[0143] In some embodiments, sequence 1 and sequence 2 are complementary to form a double-stranded region, and the 5' end of sequence 1 and the 3' end of sequence 2 are complementary to form a protruding end.

[0144] In some embodiments, the sequences shown in Formula (I) and Formula (II) are complementary to form a double-stranded region, the length of which is selected from 17 to 23 pairs of nucleotides.

[0145] In some embodiments, the sequences shown in Formula (I) and Formula (II) are complementary to form a double-stranded region, the length of which is selected from 17, 18, 19, 20, 21, 22 or 23 pairs of nucleotides, or a number of nucleotides within the range of the above values.

[0146] In some embodiments, the sequences shown in Formula (I) and Formula (II) are complementary to form a double-stranded region, the length of which is selected from 18 to 22 pairs of nucleotides or 19 to 21 pairs of nucleotides.

[0147] In some embodiments, the sequences shown in Formula (I) and Formula (II) are complementary to form a double-stranded region, the length of which is selected from 19 or 21 pairs of nucleotides.

[0148] In some embodiments, the sequences shown in formula (I) and formula (II) are complementary to form a double-stranded region, wherein N19 of the sequence shown in formula (I) is complementary to N1' of the sequence shown in formula (II).

[0149] In some embodiments, the sequences shown in formula (I) and formula (II) are complementary to form a double-stranded region, and the 3' end of the sequence shown in formula (I) and the 5' end of the sequence shown in formula (II) are complementary to form a flat end.

[0150] In some embodiments, the sequences shown in formula (I) and formula (II) are complementary to form a double-stranded region, and the 5' end of the sequence shown in formula (I) and the 3' end of the sequence shown in formula (II) are complementary to form a flat end.

[0151] In some embodiments, the sequences shown in formula (I) and formula (II) are complementary to form a double-stranded region, and the 3' end of the sequence shown in formula (I) and the 5' end of the sequence shown in formula (II) are complementary to form a protruding end.

[0152] In some embodiments, the sequences shown in formula (I) and formula (II) are complementary to form a double-stranded region, and the 5' end of the sequence shown in formula (I) and the 3' end of the sequence shown in formula (II) are complementary to form a protruding end.

[0153] Phosphate backbone modification

[0154] As used in this application, in addition to the ribosome modifications mentioned herein, the dsRNA also includes phosphate backbone modifications.

[0155] In some embodiments, the phosphate backbone modification includes phosphate ester modification of linking bonds between nucleotides and terminal phosphate ester modification.

[0156] In some embodiments, the dsRNA includes at least one linker phosphate ester modification, which includes a thiophosphate ester (PS), a dithiophosphate ester (PS2), a methylphosphonate ester (MP), a methoxypropylphosphonate ester (MOP), or an aminophosphonate ester.

[0157] In some embodiments, the linker phosphate ester modification is selected from thiophosphate ester (PS) bonds.

[0158] In some embodiments, the positive chain comprises at least one linker phosphate modification. In some embodiments, the positive chain comprises 1 to 10 linker phosphate modifications. In some embodiments, the positive chain comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 linker phosphate modifications, or a number of linker phosphate modifications within the range of the aforementioned values. In some embodiments, the positive chain comprises 1 to 4 linker phosphate modifications, 1 to 2 linker phosphate modifications, or 2 to 4 linker phosphate modifications.

[0159] In some embodiments, the antisense chain comprises at least one linker phosphate modification. In some embodiments, the antisense chain comprises 1 to 10 linker phosphate modifications. In some embodiments, the antisense chain comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 linker phosphate modifications, or a number of linker phosphate modifications within the range of the aforementioned values. In some embodiments, the antisense chain comprises 1 to 4 linker phosphate modifications, 1 to 2 linker phosphate modifications, or 2 to 4 linker phosphate modifications.

[0160] In some embodiments, the linker phosphate modification is present in the middle or at both ends of the sense and / or antisense chains.

[0161] In some embodiments, the linker phosphate modification is present between the first 6 nucleotides at the 5' end of the sense strand, between the first 6 nucleotides at the 3' end of the sense strand, between the first 6 nucleotides at the 5' end of the antisense strand, and / or between the first 6 nucleotides at the 3' end of the antisense strand.

[0162] In some embodiments, the linker phosphate modification is present between the first four nucleotides at the 5' end of the sense strand, between the first four nucleotides at the 3' end of the sense strand, between the first four nucleotides at the 5' end of the antisense strand, and / or between the first four nucleotides at the 3' end of the antisense strand.

[0163] In some embodiments, the linker phosphate modification is present at at least one of the following locations (i.e., one or more locations, for example, one, two, three, four, five, or six locations):

[0164] Between the first and second nucleotides at the 5' end of the positive strand;

[0165] Between the second and third nucleotides at the 5' end of the positive strand;

[0166] Between the first and second nucleotides at the 3' end of the positive strand;

[0167] Between the second and third nucleotides at the 3' end of the positive strand;

[0168] Between the first and second nucleotides at the 5' end of the antisense strand;

[0169] Between the second and third nucleotides at the 5' end of the antisense strand;

[0170] Between the first and second nucleotides at the 3' end of the antisense strand; or

[0171] Between the second and third nucleotides at the 3' end of the antisense strand.

[0172] In some embodiments, the linker phosphate modification is present between the first and second nucleotides at the 5' end of the sense strand, between the second and third nucleotides at the 5' end of the sense strand, between the first and second nucleotides at the 5' end of the antisense strand, between the second and third nucleotides at the 5' end of the antisense strand, between the first and second nucleotides at the 3' end of the antisense strand, and / or between the second and third nucleotides at the 3' end of the antisense strand.

[0173] In some embodiments, sequence 1 includes at least one linker phosphate modification. In some embodiments, sequence 1 includes 1 to 10 linker phosphate modifications. In some embodiments, sequence 1 includes 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 linker phosphate modifications, or a number of linker phosphate modifications within the range of the aforementioned values. In some embodiments, sequence 1 includes 1 to 4 linker phosphate modifications, 1 to 2 linker phosphate modifications, or 2 to 4 linker phosphate modifications.

[0174] In some embodiments, sequence 2 includes at least one linker phosphate modification. In some embodiments, sequence 2 includes 1 to 10 linker phosphate modifications. In some embodiments, sequence 2 includes 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 linker phosphate modifications, or a number of linker phosphate modifications within the range of the aforementioned values. In some embodiments, sequence 2 includes 1 to 4 linker phosphate modifications, 1 to 2 linker phosphate modifications, or 2 to 4 linker phosphate modifications.

[0175] In some embodiments, the linker phosphate modification is present between the first 6 nucleotides at the 5' end of sequence 1, between the first 6 nucleotides at the 3' end of sequence 1, between the first 6 nucleotides at the 5' end of sequence 2, and / or between the first 6 nucleotides at the 3' end of sequence 2.

[0176] In some embodiments, the linker phosphate modification is present between the first four nucleotides at the 5' end of sequence 1, between the first four nucleotides at the 3' end of sequence 1, between the first four nucleotides at the 5' end of sequence 2, and / or between the first four nucleotides at the 3' end of sequence 2.

[0177] In some embodiments, the linker phosphate modification is present at at least one of the following locations (i.e., one or more locations, for example, one, two, three, four, five, or six locations):

[0178] Between the first and second nucleotides at the 5' end of sequence 1;

[0179] Between the second and third nucleotides at the 5' end of sequence 1;

[0180] Between the first and second nucleotides at the 3' end of sequence 1;

[0181] Between the second and third nucleotides at the 3' end of sequence 1;

[0182] Between the first and second nucleotides at the 5' end of sequence 2;

[0183] Between the second and third nucleotides at the 5' end of sequence 2;

[0184] Between the first and second nucleotides at the 3' end of sequence 2; or

[0185] Between the second and third nucleotides at the 3' end of sequence 2.

[0186] In some embodiments, the linker phosphate modification is present between the first and second nucleotides at the 5' end of sequence 1, between the second and third nucleotides at the 5' end of sequence 1, between the first and second nucleotides at the 5' end of sequence 2, between the second and third nucleotides at the 5' end of sequence 2, between the first and second nucleotides at the 3' end of sequence 2, and / or between the second and third nucleotides at the 3' end of sequence 2.

[0187] In some embodiments, the sequence represented by formula (I) includes at least one linker phosphate modification. In some embodiments, the sequence represented by formula (I) includes 1 to 10 linker phosphate modifications. In some embodiments, the sequence represented by formula (I) includes 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 linker phosphate modifications, or a number of linker phosphate modifications within the range of the aforementioned values. In some embodiments, the sequence represented by formula (I) includes 1 to 4 linker phosphate modifications, 1 to 2 linker phosphate modifications, or 2 to 4 linker phosphate modifications.

[0188] In some embodiments, the sequence represented by formula (II) includes at least one linker phosphate modification. In some embodiments, the sequence represented by formula (II) includes 1 to 10 linker phosphate modifications. In some embodiments, the sequence represented by formula (II) includes 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 linker phosphate modifications, or a number of linker phosphate modifications within the range of the aforementioned values. In some embodiments, the sequence represented by formula (II) includes 1 to 4 linker phosphate modifications, 1 to 2 linker phosphate modifications, or 2 to 4 linker phosphate modifications.

[0189] In some embodiments, the linker phosphate modification is present between the first 6 nucleotides at the 5' end of the sequence shown in Formula (I), between the first 6 nucleotides at the 3' end of the sequence shown in Formula (I), between the first 6 nucleotides at the 5' end of the sequence shown in Formula (II), and / or between the first 6 nucleotides at the 3' end of the sequence shown in Formula (II).

[0190] In some embodiments, the linker phosphate modification is present between the first four nucleotides at the 5' end of the sequence shown in Formula (I), between the first four nucleotides at the 3' end of the sequence shown in Formula (I), between the first four nucleotides at the 5' end of the sequence shown in Formula (II), and / or between the first four nucleotides at the 3' end of the sequence shown in Formula (II).

[0191] In some embodiments, the linker phosphate modification is present at at least one of the following locations (i.e., one or more locations, for example, one, two, three, four, five, or six locations):

[0192] Between the first and second nucleotides at the 5' end of the sequence shown in formula (I);

[0193] Between the second and third nucleotides at the 5' end of the sequence shown in formula (I);

[0194] Between the first and second nucleotides at the 3' end of the sequence shown in formula (I);

[0195] Between the second and third nucleotides at the 3' end of the sequence shown in formula (I);

[0196] Between the first and second nucleotides at the 5' end of the sequence shown in formula (II);

[0197] Between the second and third nucleotides at the 5' end of the sequence shown in formula (II);

[0198] Between the first and second nucleotides at the 3' end of the sequence shown in formula (II); or

[0199] Between the second and third nucleotides at the 3' end of the sequence shown in formula (II).

[0200] In some embodiments, the linker phosphate modification is present between the first and second nucleotides at the 5' end of the sequence shown in Formula (I), between the second and third nucleotides at the 5' end of the sequence shown in Formula (I), between the first and second nucleotides at the 5' end of the sequence shown in Formula (II), between the second and third nucleotides at the 5' end of the sequence shown in Formula (II), between the first and second nucleotides at the 3' end of the sequence shown in Formula (II), and / or between the second and third nucleotides at the 3' end of the sequence shown in Formula (II).

[0201] In some embodiments, the terminal phosphate modification includes modification at the 5' or 3' end of the nucleotide.

[0202] In some embodiments, the 5' or 3' end modification of the nucleotide occurs at the 5' or 3' end of the sense strand and / or the antisense strand.

[0203] In some embodiments, the 5' end modification of the nucleotide occurs at the 5' end of either the sense or antisense strand. In some embodiments, the 5' end modification of the nucleotide occurs at the 5' end of the antisense strand.

[0204] In some embodiments, the 5' or 3' end modification of the nucleotide occurs at the 5' or 3' end of sequence 1 and / or sequence 2.

[0205] In some embodiments, the 5' end modification of the nucleotide occurs at the 5' end of sequence 1 or sequence 2. In some embodiments, the 5' end modification of the nucleotide occurs at the 5' end of sequence 2.

[0206] In some embodiments, the 5' or 3' end modification of the nucleotide occurs at the 5' or 3' end of the sequence shown in Formula (I) and / or the sequence shown in Formula (II).

[0207] In some embodiments, the 5' end modification of the nucleotide occurs at the 5' end of the sequence shown in Formula (I) or Formula (II).

[0208] In some embodiments, the terminal phosphate modification comprises nucleotide 5'-phosphate, 5'-methylphosphonate (5'-MP), 5'-thiophosphate (5'-PS), or 5'-(E)-vinylphosphonate (5'-(E)-VP) modification.

[0209] In some embodiments, the 5' end of the nucleotide comprises a 5'-(E)-vinylphosphonate.

[0210] In some embodiments, the 5' end of the nucleotide is modified with 5'-(E)-vinyl phosphate.

[0211] In some embodiments, the 5' end of the antisense chain comprises 5'-(E)-vinylphosphonate.

[0212] In some embodiments, the 5' end of the antisense chain is modified with 5'-(E)-vinyl phosphate.

[0213] In some embodiments, the 5' end of sequence 2 contains 5'-(E)-vinylphosphonate.

[0214] In some embodiments, the 5' end of sequence 2 is modified with 5'-(E)-vinyl phosphate.

[0215] In some embodiments, the 5' end of the sequence represented by formula (II) contains 5'-(E)-vinylphosphonate.

[0216] In some embodiments, the 5' end of the sequence represented by formula (II) is modified with 5'-(E)-vinyl phosphate.

[0217] Additional sequences

[0218] In some embodiments, the sense or antisense strand optionally includes an additional sequence of 1-4 nucleotides in length. In some embodiments, the additional sequence is 1, 2, 3, or 4 nucleotides in length, or a number of nucleotides within the range of these values. In some embodiments, the additional sequence is 1 or 2 nucleotides in length.

[0219] In some implementations, each nucleotide in the additional sequence is independently modified or unmodified.

[0220] In some implementations, the additional sequence is selected from unmodified or modified A, G, C, U, or T.

[0221] In some implementations, the additional sequence is selected from unmodified or modified U or T.

[0222] In some embodiments, each nucleotide in the additional sequence contains a 2'-OMe, 2'-F, or 2'-deoxy modification.

[0223] In some embodiments, the additional sequence is selected from 2'-O-methyl modified U (also known as u) or deoxythymidine nucleotide (also known as dT).

[0224] In some embodiments, when the extra sequence is two nucleotides, the extra sequence is selected from unmodified or modified UU or TT. In some embodiments, when the overhang is two nucleotides, the overhang is selected from UU or dTdT.

[0225] In some implementations, the additional sequence of the justice chain is attached to the 5' or 3' end of sequence 1.

[0226] In some implementations, the additional sequence of the antisense chain is attached to the 5' or 3' end of sequence 2.

[0227] In some implementations, the additional sequence of the justice chain is attached to the 5' or 3' end of the sequence shown in formula (I).

[0228] In some embodiments, the additional sequence of the antisense chain is attached to the 5' or 3' end of the sequence shown in formula (II).

[0229] In some implementations, the additional sequence of the justice chain may be complementary to the antisense chain to form a double-chain region, or non-complementary to form a protruding end.

[0230] In some implementations, the additional sequence of the antisense chain may be complementary to the sense chain to form a double-chain region, or non-complementary to form a protruding end.

[0231] In some implementations, the additional sequence of the antisense strand may be non-complementary, substantially complementary, or completely complementary to the target gene mRNA.

[0232] When the sense and / or antisense chains include additional sequences, and when phosphate backbone modifications are involved, refer to the detailed description of the sense and / or antisense chains in the phosphate backbone modification section above.

[0233] ligands

[0234] In some embodiments, the ligand includes a branching group and a linking group.

[0235] In some embodiments, the ligand is linked to the dsRNA via a linker group. Hereinafter, the dsRNA with the ligand linked is referred to as a dsRNA-ligand conjugate.

[0236] In some embodiments, the ligand is linked to one or more target groups via a branching group. In some embodiments, the branching group comprises a target group.

[0237] In some embodiments, the ligand contains at least one targeting group. In some embodiments, the ligand contains one, two, three, four, or five targeting groups. In some embodiments, the ligand contains two, three, or four targeting groups. In some embodiments, the ligand contains three targeting groups.

[0238] In some embodiments, the targeting group is selected from the GalNAc group.

[0239] In some embodiments, the ligand contains one, two, three, four, or five GalNAc groups. In some embodiments, the ligand contains two, three, or four GalNAc groups. In some embodiments, the ligand contains three GalNAc groups.

[0240] In some embodiments, the branching groups are selected from:

[0241] In some embodiments, the linking group is selected from:

[0242] In some embodiments, the ligand is selected from:

[0243] In some embodiments, the ligand is attached to either the sense or antisense strand. In some embodiments, the ligand is attached to the 5' or 3' end of either the sense or antisense strand. In some embodiments, the ligand is attached to the 5' or 3' end of the sense strand. In some embodiments, the ligand is attached to the 3' end of the sense strand.

[0244] In some embodiments, the ligand is linked to the sense or antisense strand of the dsRNA via a phosphate-linked structure or a thiophosphate-linked structure. In some embodiments, the ligand is linked to the sense strand via a phosphate-linked structure or a thiophosphate-linked structure. In some embodiments, the ligand is linked to the 3' end of the sense strand via a phosphate-linked structure or a thiophosphate-linked structure.

[0245] In some embodiments, the dsRNA ligand conjugate is shown below:

[0246] In some embodiments, the dsRNA ligand conjugate is selected from:

[0247] In some embodiments, the dsRNA ligand conjugate is selected from:

[0248] target genes

[0249] The dsRNA, its pharmaceutically acceptable salt, or its ligand conjugates disclosed herein may be used to treat and / or prevent diseases, conditions, or disorders associated with the target gene.

[0250] In some implementations, the disease, symptom, or disorder associated with the target gene refers to an abnormal overexpression of the target gene.

[0251] In some embodiments, the target genes include, but are not limited to: angiotensinogen (AGT), APOA, APOB, ANGPTL3, APOC3, desialyl glycoprotein receptor (ASGPR), AT3, C3, C5, Col1A1, EGFR, coagulation factor XI (FXI), F12, FVII, HBV, HCV, HDV, HIV, HSD17B13, MARC1, PD1, PD-L1, PCSK9, PKK, p53, RPTOR, PNPLA3 (Patatin-like phospholipase domain 3), SCN9A, SCD1, antithrombin III (SERPINC1), STAT3, TTR, TIMP-1, or TMPRSS6. In some embodiments, the target gene is the PNPLA3 (Patatin-like phospholipase domain 3) gene.

[0252] bases or modified bases

[0253] The nucleotides of the dsRNA in this application include natural base nucleotides, non-base nucleotides, and non-natural base nucleotides.

[0254] The nucleotides of the dsRNA in this application include natural base nucleotides.

[0255] In some embodiments, the natural base nucleotides include A, U, C, G, and T.

[0256] In some embodiments, the base-free nucleotide refers to a nucleotide that does not contain a base but only a ribose ring or a phosphate group.

[0257] In some embodiments, the non-natural base nucleotide refers to a nucleotide containing nucleobases other than adenine, guanine, cytosine, uracil, or thymine.

[0258] In some embodiments, the non-natural base nucleotides include nucleotides having a purine or pyrimidine ring structure, as well as nucleotides without a purine or pyrimidine ring structure, such as optionally substituted C5-10 aryl groups, and optionally substituted 5-10 aryl groups containing 1 to 3 heteroaryl groups selected from N, O, or S.

[0259] In some embodiments, the bases of the non-natural nucleotides include, but are not limited to: inosine, xanthine, hypoxanthine, nubularine, isoguanine, tubercidine, and substituted or modified analogs of adenine, guanine, cytosine, and uracil, such as 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, and 5-halogenated uracil. Pyrimidines and cytosines, 5-propynyluracil and cytosines, 6-azouracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thionuracil, 5-halogenated uracils, 5-(2-aminopropyl)uracil, 5-aminoallyluracil, 8-halogenated, amino, mercapto, thioalkyl, hydroxyl and other 8-substituted adenines and guanines, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine, 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O- 6-substituted purines (including 2-aminopropyladenine), 5-propynyluracil and 5-propynylcytosine, dihydrouracil, 3-deazo-5-azacytosine, 2-aminopurine, 5-alkyluracil, 7-alkylguanine, 5-alkylcytosine, 7-deazoadenine, N6,N6-dimethyladenine, 2,6-diaminopurine, 5-amino-allyl-uracil, N3-methyluracil, substituted 1,2,4-triazoles, 2-pyridones, 5-nitroindole, 3-nitropyrrole, 5-methoxy Uracil, uracil-5-hydroxyacetic acid, 5-methoxycarbonylmethyluracil, 5-methyl-2-thiouracil, 5-methoxycarbonylmethyl-2-thiouracil, 5-methylaminomethyl-2-thiouracil, 3-(3-amino-3-carboxypropyl)uracil, 3-methylcytosine, 5-methylcytosine, N4-acetylcytosine, 2-thiocytosine, N6-methyladenine, N6-isopentyladenine, 2-methylthio-N6-isopentenyladenine, N-methylguanine, or an O-alkylated base.

[0260] As used in this application, abase-free nucleotides or non-natural nucleonucleotides may be present at any position in the dsRNA. Sequence 1, the sequence shown in Formula (I), or the sense strand may contain abase-free nucleotides or non-natural nucleonucleotides, or sequence 2, the sequence shown in Formula (II), or the antisense strand may contain abase-free nucleotides or non-natural nucleonucleotides. When two or more abase-free nucleotides or non-natural nucleonucleotides are contained in the dsRNA, they may be present in sequence 1, the sequence shown in Formula (I), or the sense strand, or sequence 2, the sequence shown in Formula (II), or simultaneously in both sequence 1, the sequence shown in Formula (I), and sequence 2, the sequence shown in Formula (II).

[0261] In some embodiments, the sequence 1, the sequence shown in formula (I), or the positive strand contains one or more non-natural base nucleotides.

[0262] In some embodiments, the sequence 1, the sequence of formula (I), or the positive strand contains one, two, three, four, or five non-natural base nucleotides.

[0263] In some embodiments, the sequence 1, the sequence of formula (I), or the positive strand contains one or two non-natural base nucleotides.

[0264] In some embodiments, the sequence 2, the sequence of formula (II), or the antisense strand contains one or more non-natural base nucleotides.

[0265] In some embodiments, the sequence 2, the sequence of formula (II), or the antisense strand contains one, two, three, four, or five non-natural base nucleotides.

[0266] In some embodiments, the sequence 2, the sequence shown in formula (II), or the antisense strand contains a non-natural base nucleotide.

[0267] In some embodiments, one or more nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 of the 5' end of sequence 1 each independently comprises a non-natural base nucleotide.

[0268] In some embodiments, when the sequence 1 has 19 nucleotides, one or more nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 at the 5' end each independently contain a non-natural base nucleotide.

[0269] In some embodiments, when the sequence 1 has 19 nucleotides, one or more nucleotides at the 13th, 14th, 16th, 17th or 19th position of the 5' end each independently contain a non-natural base nucleotide.

[0270] In some embodiments, when the sequence 1 has 19 nucleotides, one or more nucleotides at the 14th, 17th, or 19th position of the 5' end each independently contain a non-natural base nucleotide.

[0271] In some embodiments, when sequence 1 has 20 nucleotides, one or more nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 at the 5' end each independently contain a non-natural base nucleotide.

[0272] In some embodiments, when the sequence 1 has 20 nucleotides, one or more nucleotides at positions 14, 15, 17, 18, or 20 at the 5' end each independently contain a non-natural base nucleotide.

[0273] In some embodiments, when the sequence 1 has 20 nucleotides, one or more nucleotides at the 15th, 18th, or 20th position of the 5' end each independently contain a non-natural base nucleotide.

[0274] In some embodiments, when the sequence 1 has 21 nucleotides, one or more nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 at the 5' end each independently contain a non-natural base nucleotide.

[0275] In some embodiments, when the sequence 1 has 21 nucleotides, one or more nucleotides at positions 15, 16, 18, 19, or 21 at the 5' end each independently contain a non-natural base nucleotide.

[0276] In some embodiments, when the sequence 1 has 21 nucleotides, one or more nucleotides at the 16th, 19th, or 21st position of the 5' end each independently contain a non-natural base nucleotide.

[0277] In some embodiments, one or more nucleotides of the sequence Na1, Na2, N1, N2, N3, N4, N5, N6, N7, N8, N9, N10, N11, N12, N13, N14, N15, N16, N17, N18 or N19 of Formula (I) each independently comprise a non-natural base nucleotide.

[0278] In some embodiments, one or more nucleotides of N13, N14, N16, N17 or N19 of the sequence shown in Formula (I) each independently contain non-natural base nucleotides.

[0279] In some embodiments, one or more nucleotides in N14, N17, or N19 of the sequence shown in Formula (I) each independently contain non-natural base nucleotides.

[0280] In some embodiments, one or more nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23 at the 5' end of sequence 2 each independently comprise a non-natural base nucleotide.

[0281] In some embodiments, when sequence 2 has 21 nucleotides, one or more nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 at the 5' end each independently contain a non-natural base nucleotide.

[0282] In some embodiments, when sequence 2 has 22 nucleotides, one or more nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 at the 5' end each independently contain a non-natural base nucleotide.

[0283] In some embodiments, when sequence 2 has 23 nucleotides, one or more nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23 at the 5' end each independently contain a non-natural base nucleotide.

[0284] In some embodiments, one or more of N1', N2', N3', N4', N5', N6', N7', N8', N9', N10', N11', N12', N13', N14', N15', N16', N17', N18', N19', N20', N21', Nb1 or Nb2 of the sequence shown in formula (II) each independently contains a non-natural base nucleotide.

[0285] In some embodiments, the bases of the non-natural nucleotide are selected from the following structure X:

[0286] In some embodiments, the non-natural nucleoside is selected from the following structures (x):

[0287] Exemplary implementation of the modification

[0288] Sequence 1 of the Justice Chain

[0289] In some embodiments, the dsRNA described in this application comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, wherein the antisense strand is at least substantially complementary to the target gene, and the sense strand comprises sequence 1.

[0290] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides with 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides with 2'-F modification.

[0291] In some embodiments, the dsRNA described in this application comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, wherein the antisense strand is at least substantially complementary to the target gene, and the antisense strand comprises sequence 2.

[0292] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification.

[0293] In some implementations, sequence 1 and sequence 2 are at least substantially complementary.

[0294] In some embodiments, unless otherwise stated, the remaining nucleotides in sequence 1 contain a 2'-OMe modification.

[0295] In some embodiments, unless otherwise stated, the remaining nucleotides in sequence 2 contain a 2'-OMe modification.

[0296] In some embodiments, sequence 1 contains two or three nucleotides with 2'-F modification, and sequence 1 does not contain three consecutive nucleotides with 2'-F modification.

[0297] In some embodiments, sequence 1 contains two or three nucleotides with 2'-F modification, sequence 1 does not contain three consecutive nucleotides with 2'-F modification, and the nucleotides of sequence 1 other than the nucleotides with 2'-F modification contain 2'-OMe modification or 2'-deoxy modification.

[0298] In some embodiments, sequence 1 contains two or three nucleotides with 2'-F modification, sequence 1 does not contain three consecutive nucleotides with 2'-F modification, and the nucleotides of sequence 1 other than the nucleotides with 2'-F modification contain 2'-OMe modification.

[0299] In some embodiments, the nucleotide containing the 2'-F modification in Sequence 1 is located at positions 5 to 11 of the 5' end of Sequence 1. In some embodiments, the nucleotide containing the 2'-F modification in Sequence 1 is located at positions 5, 6, 7, 8, 9, 10, or 11 of the 5' end of Sequence 1. In some embodiments, the nucleotide containing the 2'-F modification in Sequence 1 is located at positions 5, 7, 8, or 9 of the 5' end of Sequence 1. In some embodiments, the nucleotide containing the 2'-F modification in Sequence 1 is located at positions 6, 8, 9, or 10 of the 5' end of Sequence 1. In some embodiments, the nucleotide containing the 2'-F modification in Sequence 1 is located at positions 7, 9, 10, or 11 of the 5' end of Sequence 1. In some embodiments, the nucleotide containing the 2'-F modification in Sequence 1 is located at positions 7, 8, or 9 of the 5' end of Sequence 1. In some embodiments, the nucleotide containing the 2'-F modification in sequence 1 is located at position 8, 9, or 10 of the 5' end of sequence 1. In some embodiments, the nucleotide containing the 2'-F modification in sequence 1 is located at position 9, 10, or 11 of the 5' end of sequence 1.

[0300] In some embodiments, the 7th to 11th nucleotides at the 5' end of sequence 1 contain two nucleotides with 2'-F modification.

[0301] In some embodiments, the 7th to 11th nucleotides at the 5' end of sequence 1 contain two nucleotides with 2'-F modification, and the remaining nucleotides contain 2'-OMe modification or 2'-deoxy modification.

[0302] In some embodiments, when sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 7th, 8th and 9th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0303] In some embodiments, when sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification, 2) the 7th and 8th nucleotides at the 5' end contain a 2'-F modification, and 3) the 9th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-deoxy modification.

[0304] In some embodiments, when sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification, 2) the 7th and 9th nucleotides at the 5' end contain a 2'-F modification, and 3) the 8th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-deoxy modification.

[0305] In some embodiments, when sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification, 2) the 8th and 9th nucleotides at the 5' end contain a 2'-F modification, and 3) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-deoxy modification.

[0306] In some embodiments, when sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification, 2) two of the nucleotides at the 8th, 9th and 10th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0307] In some embodiments, when sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification, 2) the 8th and 9th nucleotides at the 5' end contain a 2'-F modification, and 3) the 10th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-deoxy modification.

[0308] In some embodiments, when sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification, 2) the 8th and 10th nucleotides at the 5' end contain a 2'-F modification, and 3) the 9th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-deoxy modification.

[0309] In some embodiments, when sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification, 2) the 9th and 10th nucleotides at the 5' end contain a 2'-F modification, and 3) the 8th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-deoxy modification.

[0310] In some embodiments, when sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 9th, 10th and 11th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0311] In some embodiments, when sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification, 2) the 9th and 10th nucleotides at the 5' end contain a 2'-F modification, and 3) the 11th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-deoxy modification.

[0312] In some embodiments, when sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification, 2) the 9th and 11th nucleotides at the 5' end contain a 2'-F modification, and 3) the 10th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-deoxy modification.

[0313] In some embodiments, when sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification, 2) the 10th and 11th nucleotides at the 5' end contain a 2'-F modification, and 3) the 9th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-deoxy modification.

[0314] In some embodiments, the 7th to 11th nucleotides at the 5' end of Sequence 1 contain one or two nucleotides containing a 2'-deoxy modification. In some embodiments, the 7th, 8th, 9th, 10th, or 11th nucleotides at the 5' end of Sequence 1 contain one or two nucleotides containing a 2'-deoxy modification.

[0315] In some embodiments, the 7th, 8th, or 9th position of the 5' end of Sequence 1 contains one or two nucleotides containing a 2'-deoxy modification. In some embodiments, the 8th, 9th, or 10th position of the 5' end of Sequence 1 contains one or two nucleotides containing a 2'-deoxy modification. In some embodiments, the 9th, 10th, or 11th position of the 5' end of Sequence 1 contains one or two nucleotides containing a 2'-deoxy modification.

[0316] In some embodiments, the 7th to 11th nucleotides at the 5' end of Sequence 1 contain a nucleotide with a 2'-deoxy modification. In some embodiments, when Sequence 1 has 19 nucleotides, the 7th to 9th nucleotides at the 5' end contain a nucleotide with a 2'-deoxy modification.

[0317] In some embodiments, when sequence 1 has 19 nucleotides, the 7th nucleotide at the 5' end contains a 2'-deoxy modification.

[0318] In some embodiments, when sequence 1 has 19 nucleotides, the 8th nucleotide at the 5' end contains a 2'-deoxy modification.

[0319] In some embodiments, when sequence 1 has 19 nucleotides, the 9th nucleotide at the 5' end contains a 2'-deoxy modification.

[0320] In some embodiments, when sequence 1 has 20 nucleotides, the 8th to 10th nucleotides at the 5' end contain a nucleotide with a 2'-deoxy modification.

[0321] In some embodiments, when sequence 1 has 20 nucleotides, the 8th nucleotide at the 5' end contains a 2'-deoxy modification.

[0322] In some embodiments, when sequence 1 has 20 nucleotides, the 9th nucleotide at the 5' end contains a 2'-deoxy modification.

[0323] In some embodiments, when sequence 1 has 20 nucleotides, the 10th nucleotide at the 5' end contains a 2'-deoxy modification.

[0324] In some embodiments, when sequence 1 has 21 nucleotides, the 9th to 11th nucleotides at the 5' end contain a nucleotide with a 2'-deoxy modification.

[0325] In some embodiments, when sequence 1 has 21 nucleotides, the 9th nucleotide at the 5' end contains a 2'-deoxy modification.

[0326] In some embodiments, when sequence 1 has 21 nucleotides, the 10th nucleotide at the 5' end contains a 2'-deoxy modification.

[0327] In some embodiments, when sequence 1 has 21 nucleotides, the 11th nucleotide at the 5' end contains a 2'-deoxy modification.

[0328] The justice chain: Na, N1, N2, N3, N4, N6, N10, N11, N12, N13, N14, N15, N16, N17, N18, and N19

[0329] In some embodiments, each of Na, N1, N2, N3, N4, N6, N10, N11, N12, N13, N14, N15, N16, N17, N18, and N19 independently represents a modified nucleotide, the modification of which does not include 2'-F modification.

[0330] In some embodiments, each of Na, N1, N2, N3, N4, N6, N10, N11, N12, N13, N14, N15, N16, N17, N18, and N19 independently represents a modified nucleotide, the modification comprising a 2'-OMe modification.

[0331] Justice Chain N5

[0332] In some embodiments, N5 includes a 2'-F modification.

[0333] In some embodiments, N5 includes a 2'-OMe modification.

[0334] Justice Chain N7, N8 and N9

[0335] In some embodiments, N7, N8, and N9 each independently represent a modified nucleotide, the modification including 2'-OMe modification, 2-F modification, or 2'-deoxy modification, and the modifications of N7, N8, and N9 are not all selected from 2'-OMe modification or 2'-F modification.

[0336] In some embodiments, at least one (e.g., one or both) of N7, N8 and N9 contains 2'-F modification, and each of the others independently contains 2'-OMe modification or 2'-deoxygenation modification.

[0337] In some embodiments, one of N7, N8 and N9 contains a 2'-F modification, and each of the others independently contains a 2'-OMe modification or a 2'-deoxygenation modification.

[0338] In some embodiments, N7 includes 2'-F modification, and N8 and N9 each independently include 2'-OMe modification or 2'-deoxygenation modification.

[0339] In some embodiments, N7 includes a 2'-F modification, N8 includes a 2'-OMe modification, and N9 includes a 2'-OMe modification.

[0340] In some embodiments, N7 contains 2'-F modification, N8 contains 2'-deoxygenation modification, and N9 contains 2'-deoxygenation modification.

[0341] In some embodiments, N7 contains 2'-F modification, N8 contains 2'-OMe modification, and N9 contains 2'-deoxygenation modification.

[0342] In some embodiments, N7 contains 2'-F modification, N8 contains 2'-deoxygenation modification, and N9 contains 2'-OMe modification.

[0343] In some embodiments, N8 includes 2'-F modification, and N7 and N9 each independently include 2'-OMe modification or 2'-deoxygenation modification.

[0344] In some embodiments, N8 includes a 2'-F modification, N7 includes a 2'-OMe modification, and N9 includes a 2'-OMe modification.

[0345] In some embodiments, N8 contains 2'-F modification, N7 contains 2'-deoxygenation modification, and N9 contains 2'-deoxygenation modification.

[0346] In some embodiments, N8 contains 2'-F modification, N7 contains 2'-OMe modification, and N9 contains 2'-deoxygenation modification.

[0347] In some embodiments, N8 contains 2'-F modification, N7 contains 2'-deoxygenation modification, and N9 contains 2'-OMe modification.

[0348] In some embodiments, N9 includes a 2'-F modification, and N7 and N8 each independently include a 2'-OMe modification or a 2'-deoxygenation modification.

[0349] In some embodiments, N9 includes a 2'-F modification, N7 includes a 2'-OMe modification, and N8 includes a 2'-OMe modification.

[0350] In some embodiments, N9 contains 2'-F modification, N7 contains 2'-deoxygenation modification, and N8 contains 2'-deoxygenation modification.

[0351] In some embodiments, N9 contains 2'-F modification, N7 contains 2'-OMe modification, and N8 contains 2'-deoxygenation modification.

[0352] In some embodiments, N9 contains 2'-F modification, N7 contains 2'-deoxygenation modification, and N8 contains 2'-OMe modification.

[0353] In some embodiments, two of N7, N8 and N9 contain 2'-F, and each of the remaining ones contains 2'-OMe modification or 2'-deoxygenation modification.

[0354] In some embodiments, N7 and N8 contain 2'-F modification, and N9 contains 2'-OMe modification or 2'-deoxygenation modification.

[0355] In some embodiments, N7 and N8 contain 2'-F modifications, and N9 contains 2'-OMe modifications.

[0356] In some embodiments, N7 and N8 contain 2'-F modification, and N9 contains 2'-deoxygenation modification.

[0357] In some embodiments, N7 and N9 contain 2'-F modification, and N8 contains 2'-OMe modification or 2'-deoxygenation modification.

[0358] In some embodiments, N7 and N9 contain 2'-F modifications, and N8 contains 2'-OMe modifications.

[0359] In some embodiments, N7 and N9 contain 2'-F modification, and N8 contains 2'-deoxygenation modification.

[0360] In some embodiments, N8 and N9 contain 2'-F modification, and N7 contains 2'-OMe modification or 2'-deoxygenation modification.

[0361] In some embodiments, N8 and N9 contain 2'-F modifications, and N7 contains 2'-OMe modifications.

[0362] In some embodiments, N8 and N9 contain 2'-F modification, and N7 contains 2'-deoxygenation modification.

[0363] Justice chain variables Na and x

[0364] In some implementations, x is selected from 0.

[0365] In some embodiments, x is selected from 1, then Na represents a modified nucleotide Na1.

[0366] In some embodiments, x is selected from 2, then Na represents two modified nucleotides Na1Na2, wherein Na1 is linked to N1.

[0367] Sequence 2 of the antisense chain

[0368] In some embodiments, sequence 2 contains two, three, or four nucleotides containing a 2'-F modification. In some embodiments, the nucleotides containing a 2'-F modification are located at positions 2, 6, 14, or 16 of the 5' end of sequence 2. In some embodiments, the nucleotides containing a 2'-F modification are located at positions 2 and 14 of the 5' end of sequence 2, and position 6 or 16 of the 5' end optionally contains a 2'-F modification, a 2'-OMe modification, or a 2'-deoxy modification. In some embodiments, the nucleotides containing a 2'-F modification are located at positions 2, 14, and 16 of the 5' end of sequence 2, and position 6 of the 5' end optionally contains a 2'-F modification, a 2'-OMe modification, or a 2'-deoxy modification.

[0369] In some embodiments, sequence 2 contains two, three, or four nucleotides containing 2'-F modification, and the nucleotides of sequence 2 other than the nucleotides containing 2'-F modification contain 2'-OMe modification or 2'-deoxy modification.

[0370] In some embodiments, sequence 2 contains two, three, or four nucleotides containing 2'-F modification, and the nucleotides of sequence 2 other than the nucleotides containing 2'-F modification contain 2'-OMe modification.

[0371] In some embodiments, the nucleotides at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, and 13 at the 5' end of sequence 2 each independently contain a 2'-OMe modification.

[0372] In some embodiments, the nucleotides at positions 2 and 14 of the 5' end of sequence 2 contain a 2'-F modification.

[0373] In some embodiments, the 6th nucleotide at the 5' end of sequence 2 contains a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification. In some embodiments, the 6th nucleotide at the 5' end of sequence 2 contains a 2'-OMe modification or a 2'-deoxy modification. In some embodiments, the 6th nucleotide at the 5' end of sequence 2 contains a 2'-F modification or a 2'-deoxy modification.

[0374] In some embodiments, the 6th nucleotide at the 5' end of sequence 2 contains a 2'-F modification.

[0375] In some embodiments, the 6th nucleotide at the 5' end of sequence 2 does not contain a 2'-F modification, but contains a 2'-OMe modification or a 2'-deoxy modification. In some embodiments, the 6th nucleotide at the 5' end of sequence 2 contains a 2'-OMe modification. In some embodiments, the 6th nucleotide at the 5' end of sequence 2 contains a 2'-deoxy modification.

[0376] In some embodiments, the 5' end 16th nucleotide of sequence 2 contains a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification. In some embodiments, the 5' end 16th nucleotide of sequence 2 contains a 2'-F modification.

[0377] In some embodiments, one to three nucleotides at positions 15 to 19, 15 to 20, 15 to 21, 15 to 22, or 15 to 23 of the 5' end of sequence 2 contain a 2'-deoxy modification.

[0378] In some embodiments, one of the nucleotides at positions 15 to 19, 15 to 20, 15 to 21, 15 to 22, or 15 to 23 at the 5' end of sequence 2 contains a 2'-deoxy modification.

[0379] In some embodiments, one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end of sequence 2 contains a 2'-deoxy modification.

[0380] In some embodiments, one of the nucleotides at positions 15, 17, 18, 19, 20, 21, 22, or 23 of the 5' end of sequence 2 contains a 2'-deoxy modification.

[0381] In some embodiments, sequence 2: 1) the nucleotides at positions 2, 14, and 16 of the 5' end contain 2'-F modification, and 2) the nucleotide at position 6 of the 5' end does not contain 2'-F modification, but contains 2'-OMe modification or 2'-deoxy modification.

[0382] In some embodiments, sequence 2 includes: 1) nucleotides 2, 14, and 16 at the 5' end containing 2'-F modification; 2) nucleotide 6 at the 5' end not containing 2'-F modification, but containing 2'-OMe modification or 2'-deoxy modification; and 3) the remaining nucleotides of sequence 2 containing 2'-OMe modification.

[0383] In some embodiments, sequence 2 includes: 1) nucleotides at positions 2, 6, and 14 of the 5' end containing a 2'-F modification; 2) nucleotide at position 16 of the 5' end containing a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification; and 3) one of the nucleotides at positions 15 to 19, 15 to 20, 15 to 21, 15 to 22, or 15 to 23 of the 5' end containing a 2'-deoxy modification.

[0384] In some embodiments, sequence 2 includes: 1) nucleotides at positions 2, 6, and 14 of the 5' end containing a 2'-F modification; 2) nucleotide at position 16 of the 5' end containing a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification; 3) one nucleotide at positions 15 to 19, 15 to 20, 15 to 21, 15 to 22, or 15 to 23 of the 5' end containing a 2'-deoxy modification; and 4) the remaining nucleotides of sequence 2 containing a 2'-OMe modification.

[0385] In some embodiments, sequence 2 includes: 1) nucleotides at positions 2, 6, and 14 of the 5' end containing a 2'-F modification; 2) nucleotide at position 16 of the 5' end containing a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification; and 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end containing a 2'-deoxy modification.

[0386] In some embodiments, sequence 2 includes: 1) nucleotides at positions 2, 6, and 14 of the 5' end containing a 2'-F modification; 2) nucleotide at position 16 of the 5' end containing a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end containing a 2'-deoxy modification; and 4) the remaining nucleotides of sequence 2 containing a 2'-OMe modification.

[0387] In some embodiments, sequence 2 includes: 1) nucleotides 2 and 14 at the 5' end containing 2'-F modification; 2) nucleotide 6 at the 5' end not containing 2'-F modification, but containing 2'-OMe modification or 2'-deoxy modification; 3) nucleotide 16 at the 5' end containing 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; and 4) one of the nucleotides at positions 15 to 19, 15 to 20, 15 to 21, 15 to 22, or 15 to 23 at the 5' end containing 2'-deoxy modification.

[0388] In some embodiments, sequence 2 includes: 1) nucleotides 2 and 14 at the 5' end containing 2'-F modification; 2) nucleotide 6 at the 5' end not containing 2'-F modification, but containing 2'-OMe modification or 2'-deoxy modification; 3) nucleotide 16 at the 5' end containing 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one nucleotide at positions 15 to 19, 15 to 20, 15 to 21, 15 to 22, or 15 to 23 at the 5' end containing 2'-deoxy modification; and 5) the remaining nucleotides of sequence 2 containing 2'-OMe modification.

[0389] In some embodiments, sequence 2 includes: 1) nucleotides 2 and 14 at the 5' end containing 2'-F modification; 2) nucleotide 6 at the 5' end not containing 2'-F modification, but containing 2'-OMe modification or 2'-deoxy modification; 3) nucleotide 16 at the 5' end containing 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; and 4) one of the nucleotides at the 5' end, either 15, 16, 17, 18, 19, 20, 21, 22, or 23, containing 2'-deoxy modification.

[0390] In some embodiments, sequence 2 includes: 1) nucleotides at positions 2 and 14 of the 5' end containing 2'-F modification; 2) nucleotide at position 6 of the 5' end not containing 2'-F modification, but containing 2'-OMe modification or 2'-deoxy modification; 3) nucleotide at position 16 of the 5' end containing 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end containing 2'-deoxy modification; and 5) the remaining nucleotides of sequence 2 containing 2'-OMe modification.

[0391] In some embodiments, the 6th or 15th nucleotide from the 5' end of sequence 2 to the last nucleotide contains one or two nucleotides containing a 2'-deoxy modification.

[0392] In some embodiments, the 6th or 15th nucleotide from the 5' end of sequence 2 to the last nucleotide (e.g., the 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, or 23rd nucleotide) contains a nucleotide with a 2'-deoxy modification.

[0393] In some embodiments, the 6th nucleotide at the 5' end or the last nucleotide at the 5' end of sequence 2 contains one or two nucleotides containing a 2'-deoxy modification.

[0394] In some embodiments, the 6th nucleotide at the 5' end of sequence 2 contains a 2'-deoxy modification.

[0395] In some embodiments, the 15th nucleotide at the 5' end of sequence 2 up to the last nucleotide contains one or two nucleotides containing a 2'-deoxy modification.

[0396] In some embodiments, the nucleotides from the 15th to the last (e.g., the 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, or 23rd) of the 5' end of sequence 2 contain a nucleotide with a 2'-deoxy modification.

[0397] In some embodiments, the last nucleotide at the 5' end of sequence 2 contains a 2'-deoxy modification.

[0398] In some embodiments, one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 at the 5' end of sequence 2 contains a 2'-deoxy modification.

[0399] In some embodiments, when sequence 2 has 21 nucleotides, one of the nucleotides at positions 15, 16, 17, 18, 19, 20, or 21 at the 5' end of sequence 2 contains a 2'-deoxy modification.

[0400] In some embodiments, when sequence 2 has 22 nucleotides, one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, or 22 at the 5' end of sequence 2 contains a 2'-deoxy modification.

[0401] In some embodiments, when sequence 2 has 23 nucleotides, one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 at the 5' end of sequence 2 contains a 2'-deoxy modification.

[0402] In some embodiments, when sequence 2 has 21 nucleotides, the 21st nucleotide at the 5' end of sequence 2 contains a 2'-deoxy modification.

[0403] In some embodiments, when sequence 2 has 22 nucleotides, the 22nd nucleotide at the 5' end of sequence 2 contains a 2'-deoxy modification.

[0404] In some embodiments, when sequence 2 has 23 nucleotides, the 23rd nucleotide at the 5' end of sequence 2 contains a 2'-deoxy modification.

[0405] Antonyms N1', N3', N4', N5', N7', N8', N9', N10', N11', N12', and N13'

[0406] In some embodiments, N1', N3', N4', N5', N7', N8', N9', N10', N11', N12', and N13' each independently represent a modified nucleotide, the modification of which does not include 2'-F modification.

[0407] In some embodiments, N1', N3', N4', N5', N7', N8', N9', N10', N11', N12', and N13' each independently represent a modified nucleotide, the modification including a 2'-OMe modification.

[0408] antisense chains N2' and N14'

[0409] In some embodiments, N2' and N14' each independently represent a modified nucleotide, the modification of which does not include 2'-OMe modification.

[0410] In some embodiments, N2' and N14' each independently represent a modified nucleotide, the modification including a 2'-F modification.

[0411] antisense chain N6'

[0412] In some implementations, N6' includes a 2'-F modification.

[0413] In some embodiments, the N6' includes 2'-OMe modification or 2'-deoxygenation modification.

[0414] In some embodiments, N6' includes a 2'-OMe modification.

[0415] In some embodiments, the N6' includes a 2'-deoxygenation modification.

[0416] The antisense chains N15', N16', N17', N18', N19', N20', N21' and each Nb

[0417] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including 2'-OMe modification.

[0418] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide that does not contain 2'-deoxy modification.

[0419] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, at least one of which contains a 2'-deoxy modification.

[0420] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein three, two or one nucleotide contain a 2'-deoxy modification.

[0421] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein two or one nucleotide contains a 2'-deoxy modification.

[0422] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein two nucleotides contain a 2'-deoxy modification.

[0423] In some embodiments, N15', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification; N16' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification or 2'-deoxy modification; wherein, a nucleotide includes 2'-deoxy modification.

[0424] In some embodiments, N15', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification; N16' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification or 2'-deoxy modification; wherein, two nucleotides contain 2'-deoxy modification.

[0425] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein two nucleotides contain a 2'-deoxy modification; except for the nucleotides containing a 2'-deoxy modification, each of the remaining N15', N17', N18', N19', N20', N21' and each Nb each independently contains a 2'-OMe modification, and the remaining N16' contains either a 2'-OMe modification or a 2'-F modification.

[0426] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein one of the nucleotides contains a 2'-deoxy modification; except for the nucleotides containing a 2'-deoxy modification, each of the remaining N15', N17', N18', N19', N20', N21' and each Nb each independently contains a 2'-OMe modification, and the remaining N16' contains either a 2'-OMe modification or a 2'-F modification.

[0427] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein only N15' contains a 2'-deoxy modification.

[0428] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein N15' contains a 2'-deoxy modification, N16' contains a 2'-OMe modification or a 2'-F modification, and N17', N18', N19', N20', N21' and each Nb contain a 2'-OMe modification. In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein N15' contains a 2'-deoxy modification, N16' contains a 2'-F modification, and N17', N18', N19', N20', N21' and each Nb contain a 2'-OMe modification.

[0429] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein only N16' contains a 2'-deoxy modification.

[0430] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein N16' contains a 2'-deoxy modification, and N15', N17', N18', N19', N20', N21' and each Nb contain a 2'-OMe modification.

[0431] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein only N17' contains a 2'-deoxy modification.

[0432] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein N17' contains a 2'-deoxy modification, N15', N18', N19', N20', N21' and each Nb contain a 2'-OMe modification, and N16' contains either a 2'-OMe modification or a 2'-F modification. In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein N17' contains a 2'-deoxy modification, N15', N18', N19', N20', N21' and each Nb contain a 2'-OMe modification, and N16' contains a 2'-F modification.

[0433] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein only N18' contains a 2'-deoxy modification.

[0434] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein N18' contains a 2'-deoxy modification, N15', N17', N19', N20', N21' and each Nb contain a 2'-OMe modification, and N16' contains either a 2'-OMe modification or a 2'-F modification. In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein N18' contains a 2'-deoxy modification, N15', N17', N19', N20', N21' and each Nb contain a 2'-OMe modification, and N16' contains a 2'-F modification.

[0435] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein only N19' contains a 2'-deoxy modification.

[0436] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein N19' contains a 2'-deoxy modification, N15', N17', N18', N20', N21' and each Nb contain a 2'-OMe modification, and N16' contains either a 2'-OMe modification or a 2'-F modification. In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein N19' contains a 2'-deoxy modification, N15', N17', N18', N20', N21' and each Nb contain a 2'-OMe modification, and N16' contains a 2'-F modification.

[0437] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein only N20' contains a 2'-deoxy modification.

[0438] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein N20' contains a 2'-deoxy modification, N15', N17', N18', N19', N21' and each Nb contain a 2'-OMe modification, and N16' contains either a 2'-OMe modification or a 2'-F modification. In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb each independently represent a modified nucleotide, wherein N20' contains a 2'-deoxy modification, N15', N17', N18', N19', N21' and each Nb contain a 2'-OMe modification, and N16' contains a 2'-F modification.

[0439] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein only N21' contains a 2'-deoxy modification.

[0440] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein N21' contains a 2'-OMe modification or a 2'-deoxy modification, N15', N17', N18', N19', N20' and each Nb contain a 2'-OMe modification, and N16' contains a 2'-OMe modification or a 2'-F modification. In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein N21' contains a 2'-deoxy modification, N15', N17', N18', N19', N20' and each Nb contain a 2'-OMe modification, and N16' contains a 2'-F modification.

[0441] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein only one nucleotide in Nb contains a 2'-deoxy modification.

[0442] In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein one nucleotide in Nb contains a 2'-deoxy modification, N15', N17', N18', N19', N20', N21' and each of the remaining Nb contain a 2'-OMe modification, and N16' contains either a 2'-OMe modification or a 2'-F modification. In some embodiments, N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, wherein one nucleotide in Nb contains a 2'-deoxy modification, N15', N17', N18', N19', N20', N21' and each of the remaining Nb contain a 2'-OMe modification, and N16' contains a 2'-F modification.

[0443] antisense chain N16'

[0444] In some implementations, N16' includes a 2'-OMe modification or a 2'-F modification.

[0445] In some implementations, N16' includes a 2'-OMe modification.

[0446] In some implementations, N16' includes a 2'-F modification.

[0447] In some embodiments, the N16' contains a 2'-deoxygenation modification.

[0448] antisense chain Nb

[0449] In some implementations, y is selected from 0.

[0450] In some embodiments, y is selected from 1, then Nb represents a modified nucleotide Nb1.

[0451] In some embodiments, when y is selected from 1, Nb1 contains 2'-deoxygenation modification, N15', N17', N18', N19', N20' and N21' contain 2'-OMe modification, and N16' contains 2'-F modification.

[0452] In some embodiments, y is selected from 2, then Nb represents two modified nucleotides Nb1Nb2, wherein Nb1 is connected to N21'.

[0453] In some embodiments, when y is selected from 2, Nb1 and Nb2 contain 2'-OMe modification, and one of N15', N16', N17', N18', N19', N20' and N21' contains 2'-deoxygenation modification.

[0454] In some embodiments, when y is selected from 2, Nb1 and Nb2 contain 2'-OMe modification, and N15', N17', N18', N19', N20', and N21' each independently represent a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification; N16' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; wherein, one of N15', N16', N17', N18', N19', N20', and N21' contains 2'-deoxy modification.

[0455] In some embodiments, when y is selected from 2, Nb1 and Nb2 contain 2'-OMe modification, one of N15', N16', N17', N18', N19', N20' and N21' contains 2'-deoxy modification, the remaining N15', N17', N18', N19', N20' and N21' contain 2'-OMe modification, and the remaining N16' contains either 2'-OMe modification or 2'-F modification.

[0456] In some embodiments, when y is selected from 2, Nb1 contains 2'-deoxygenation modification, N15', N17', N18', N19', N20', N21' and Nb2 contain 2'-OMe modification, and N16' contains 2'-F modification.

[0457] In some embodiments, when y is selected from 2, Nb2 contains 2'-deoxygenation modification, N15', N17', N18', N19', N20', N21' and Nb1 contain 2'-OMe modification, and N16' contains 2'-F modification.

[0458] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, the sequence 1 and sequence 2 being at least substantially complementary, and each nucleotide of the sequence 1 and sequence 2 being a modified nucleotide;

[0459] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification;

[0460] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification;

[0461] When the sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 7th, 8th and 9th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0462] When the sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 8th, 9th and 10th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0463] When the sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 9th, 10th and 11th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0464] Sequence 2: 1) The nucleotides at positions 2 and 14 of the 5' end contain a 2'-F modification; 2) The nucleotide at position 6 of the 5' end contains a 2'-OMe modification or a 2'-deoxy modification; 3) The nucleotide at position 16 contains a 2'-F modification, a 2'-OMe modification, or a 2'-deoxy modification; or,

[0465] Sequence 2: 1) The nucleotides at positions 2, 14, and 16 of the 5' end contain 2'-F modification, and 2) The nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification.

[0466] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, the sequence 1 and sequence 2 being at least substantially complementary, and each nucleotide of the sequence 1 and sequence 2 being a modified nucleotide;

[0467] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification;

[0468] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification;

[0469] When the sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 7th, 8th and 9th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0470] When the sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 8th, 9th and 10th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0471] When the sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 9th, 10th and 11th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0472] Sequence 2: 1) The nucleotides at positions 2 and 14 of the 5' end contain a 2'-F modification; 2) The nucleotide at position 6 of the 5' end contains a 2'-OMe modification or a 2'-deoxy modification; 3) The nucleotide at position 16 contains a 2'-F modification, a 2'-OMe modification, or a 2'-deoxy modification; or,

[0473] Sequence 2: 1) The nucleotides at positions 2, 14, and 16 of the 5' end contain a 2'-F modification, and 2) The nucleotide at position 6 of the 5' end contains a 2'-OMe modification or a 2'-deoxy modification;

[0474] The remaining nucleotides of sequence 1 and sequence 2 contain 2'-OMe modification.

[0475] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, the sequence 1 and sequence 2 being at least substantially complementary, and each nucleotide of the sequence 1 and sequence 2 being a modified nucleotide;

[0476] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification;

[0477] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification;

[0478] When the sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 7th, 8th and 9th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0479] When the sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 8th, 9th and 10th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0480] When the sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 9th, 10th and 11th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0481] When sequence 2 has 21 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, or 21 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-F modification.

[0482] When sequence 2 has 22 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, or 22 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-F modification.

[0483] When sequence 2 has 23 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-F modification.

[0484] or,

[0485] When sequence 2 has 21 nucleotides: 1) the nucleotides at positions 2, 6, and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, or 21 of the 5' end contains 2'-deoxy modification.

[0486] When sequence 2 has 22 nucleotides: 1) the nucleotides at positions 2, 6, and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, or 22 of the 5' end contains 2'-deoxy modification.

[0487] When sequence 2 has 23 nucleotides: 1) the nucleotides at positions 2, 6, and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end contains 2'-deoxy modification.

[0488] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, the sequence 1 and sequence 2 being at least substantially complementary, and each nucleotide of the sequence 1 and sequence 2 being a modified nucleotide;

[0489] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification;

[0490] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification;

[0491] When the sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 7th, 8th and 9th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0492] When the sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 8th, 9th and 10th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0493] When the sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 9th, 10th and 11th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0494] When sequence 2 has 21 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, or 21 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-F modification.

[0495] When sequence 2 has 22 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, or 22 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-F modification.

[0496] When sequence 2 has 23 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-F modification.

[0497] or,

[0498] When sequence 2 has 21 nucleotides: 1) the nucleotides at positions 2, 6, and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, or 21 of the 5' end contains 2'-deoxy modification.

[0499] When sequence 2 has 22 nucleotides: 1) the nucleotides at positions 2, 6, and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, or 22 of the 5' end contains 2'-deoxy modification.

[0500] When sequence 2 has 23 nucleotides: 1) the nucleotides at positions 2, 6, and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end contains 2'-deoxy modification.

[0501] The remaining nucleotides of sequence 1 and sequence 2 contain 2'-OMe modification.

[0502] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, the sequence 1 and sequence 2 being at least substantially complementary, and each nucleotide of the sequence 1 and sequence 2 being a modified nucleotide;

[0503] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification;

[0504] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification;

[0505] When the sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 7th, 8th and 9th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0506] When the sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 8th, 9th and 10th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0507] When the sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 9th, 10th and 11th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0508] When sequence 2 has 21 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification; 3) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, or 21 of the 5' end contains 2'-deoxy modification.

[0509] When sequence 2 has 22 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification; 3) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, or 22 of the 5' end contains 2'-deoxy modification.

[0510] When sequence 2 has 23 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification; 3) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end contains 2'-deoxy modification.

[0511] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA) of a pharmaceutically acceptable salt or a ligand conjugate thereof, the dsRNA comprising a sense strand and an antisense strand, each strand having 14 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, the sequence 1 and sequence 2 being at least substantially complementary, and each nucleotide of the sequence 1 and sequence 2 being a modified nucleotide;

[0512] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification;

[0513] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification;

[0514] When the sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 7th, 8th and 9th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0515] When the sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 8th, 9th and 10th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0516] When the sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 9th, 10th and 11th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification.

[0517] When sequence 2 has 21 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification; 3) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, or 21 of the 5' end contains 2'-deoxy modification.

[0518] When sequence 2 has 22 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification; 3) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, or 22 of the 5' end contains 2'-deoxy modification.

[0519] When sequence 2 has 23 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification; 3) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end contains 2'-deoxy modification.

[0520] The remaining nucleotides of sequence 1 and sequence 2 contain 2'-OMe modification.

[0521] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising the sequence shown in formula (I), and the antisense strand comprising the sequence shown in formula (II).

[0522] Formula (I): 5'-(Na) x N1N2N3N4N5N6N7N8N9N10N11N12N13N14N15N16N17N18N19-3',

[0523] Formula (II): 5'-N1'N2'N3'N4'N5'N6'N7'N8'N9'N10'N11'N12'N13'N14'N15'N16'N17'N18'N19'N20'N21'(Nb) y -3';

[0524] Each of Na, N1, N2, N3, N4, N6, N10, N11, N12, N13, N14, N15, N16, N17, N18, and N19 independently represents a modified nucleotide, the modification including a 2'-OMe modification.

[0525] The N5 represents a modified nucleotide, the modification comprising either a 2'-OMe modification or a 2'-F modification.

[0526] N7, N8, and N9 each independently represent a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification, wherein two of N7, N8, and N9 contain 2'-F modification, and each of the remaining ones contains either 2'-OMe modification or 2'-deoxy modification.

[0527] The x is selected from 0, 1, or 2; when x is selected from 1, Na represents a modified nucleotide Na1; when x is selected from 2, Na represents two modified nucleotides Na1Na2, wherein Na1 is linked to N1.

[0528] N1', N3', N4', N5', N7', N8', N9', N10', N11', N12', and N13' each independently represent a modified nucleotide, the modification including a 2'-OMe modification.

[0529] N2' and N14' each independently represent a modified nucleotide, the modification including a 2'-F modification.

[0530] The N6' represents a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification.

[0531] The N15', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification.

[0532] The N16' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification.

[0533] or,

[0534] The N15', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including a 2'-OMe modification.

[0535] The N16' represents a modified nucleotide, the modification including 2'-OMe modification or 2'-F modification.

[0536] The y is selected from 0, 1 or 2; when y is selected from 1, Nb represents a modified nucleotide Nb1; when y is selected from 2, Nb represents two modified nucleotides Nb1Nb2, wherein Nb1 is connected to N21'.

[0537] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising the sequence shown in formula (I), and the antisense strand comprising the sequence shown in formula (II).

[0538] Formula (I): 5'-(Na)xN1N2N3N4N5N6N7N8N9N10N11N12N13N14N15N16N17N18N19-3',

[0539] Formula (II): 5'-N1'N2'N3'N4'N5'N6'N7'N8'N9'N10'N11'N12'N13'N14'N15'N16'N17'N18'N19'N20'N21'(Nb)y-3';

[0540] Each of Na, N1, N2, N3, N4, N6, N10, N11, N12, N13, N14, N15, N16, N17, N18, and N19 independently represents a modified nucleotide, the modification including a 2'-OMe modification.

[0541] The N5 represents a modified nucleotide, the modification comprising either a 2'-OMe modification or a 2'-F modification.

[0542] N7, N8, and N9 each independently represent a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification, wherein two of N7, N8, and N9 contain 2'-F, and each of the remaining ones contains either 2'-OMe modification or 2'-deoxy modification.

[0543] The x is selected from 0, 1, or 2; when x is selected from 1, Na represents a modified nucleotide Na1; when x is selected from 2, Na represents two modified nucleotides Na1Na2, wherein Na1 is linked to N1.

[0544] N1', N3', N4', N5', N7', N8', N9', N10', N11', N12', and N13' each independently represent a modified nucleotide, the modification including a 2'-OMe modification.

[0545] N2' and N14' each independently represent a modified nucleotide, the modification including a 2'-F modification.

[0546] The N6' represents a modified nucleotide, the modification comprising a 2'-F modification.

[0547] N15', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification; N16' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; wherein, a nucleotide includes 2'-deoxy modification, or,

[0548] Each of N15', N16', N17', N18', N19', N20', N21' and each Nb independently represents a modified nucleotide, wherein one of the nucleotides contains a 2'-deoxy modification. Except for the nucleotides containing a 2'-deoxy modification, each of the remaining N15', N17', N18', N19', N20', N21' and each Nb independently contains a 2'-OMe modification, and the remaining N16' contains either a 2'-OMe modification or a 2'-F modification.

[0549] The y is selected from 0, 1 or 2; when y is selected from 1, Nb represents a modified nucleotide Nb1; when y is selected from 2, Nb represents two modified nucleotides Nb1Nb2, wherein Nb1 is connected to N21'.

[0550] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising the sequence shown in formula (I), and the antisense strand comprising the sequence shown in formula (II).

[0551] Formula (I): 5'-(Na)xN1N2N3N4N5N6N7N8N9N10N11N12N13N14N15N16N17N18N19-3',

[0552] Formula (II): 5'-N1'N2'N3'N4'N5'N6'N7'N8'N9'N10'N11'N12'N13'N14'N15'N16'N17'N18'N19'N20'N21'(Nb)y-3';

[0553] Each of Na, N1, N2, N3, N4, N6, N10, N11, N12, N13, N14, N15, N16, N17, N18, and N19 independently represents a modified nucleotide, the modification including a 2'-OMe modification.

[0554] The N5 represents a modified nucleotide, the modification comprising either a 2'-OMe modification or a 2'-F modification.

[0555] N7, N8, and N9 each independently represent a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification, wherein two of N7, N8, and N9 contain 2'-F, and each of the remaining ones contains either 2'-OMe modification or 2'-deoxy modification.

[0556] The x is selected from 0, 1, or 2; when x is selected from 1, Na represents a modified nucleotide Na1; when x is selected from 2, Na represents two modified nucleotides Na1Na2, wherein Na1 is linked to N1.

[0557] N1', N3', N4', N5', N7', N8', N9', N10', N11', N12', and N13' each independently represent a modified nucleotide, the modification including a 2'-OMe modification.

[0558] N2' and N14' each independently represent a modified nucleotide, the modification including a 2'-F modification.

[0559] The N6' represents a modified nucleotide, the modification comprising a 2'-OMe modification or a 2'-deoxy modification.

[0560] N15', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification; N16' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; wherein, a nucleotide includes 2'-deoxy modification, or,

[0561] Each of N15', N16', N17', N18', N19', N20', N21' and each Nb independently represents a modified nucleotide, wherein one of the nucleotides contains a 2'-deoxy modification. Except for the nucleotides containing a 2'-deoxy modification, each of the remaining N15', N17', N18', N19', N20', N21' and each Nb independently contains a 2'-OMe modification, and the remaining N16' contains either a 2'-OMe modification or a 2'-F modification.

[0562] The y is selected from 0, 1 or 2; when y is selected from 1, Nb represents a modified nucleotide Nb1; when y is selected from 2, Nb represents two modified nucleotides Nb1Nb2, wherein Nb1 is connected to N21'.

[0563] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, said dsRNA comprising a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, said antisense strand being at least substantially complementary to a target gene, said sense strand comprising the sequence shown in formula (I), said antisense strand comprising the sequence shown in formula (II), said formulas (I) and (II) being selected from the following groups in Table 1:

[0564] Table 1

[0565] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, said dsRNA comprising a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, said antisense strand being at least substantially complementary to a target gene, said sense strand comprising the sequence shown in formula (I), said antisense strand comprising the sequence shown in formula (II), said formulas (I) and (II) being selected from the following groups in Table 2:

[0566] Table 2

[0567] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, said dsRNA comprising a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, said antisense strand being at least substantially complementary to a target gene, said sense strand comprising the sequence shown in formula (I), said antisense strand comprising the sequence shown in formula (II), said formulas (I) and (II) being selected from the following groups in Table 3:

[0568] Table 3

[0569] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, said dsRNA comprising a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, said antisense strand being at least substantially complementary to a target gene, said sense strand comprising the sequence shown in formula (I), said antisense strand comprising the sequence shown in formula (II), said formulas (I) and (II) being selected from the following groups in Table 4:

[0570] Table 4

[0571] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, the antisense strand being at least substantially complementary to a target gene, and the sense strand or antisense strand being selected from the groups listed in Table 5 below:

[0572] Table 5

[0573] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, the antisense strand being at least substantially complementary to a target gene, and the sense strand or antisense strand being selected from the groups listed in Table 6 below:

[0574] Table 6

[0575] Modification implementation methods containing non-natural base X

[0576] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, the sequence 1 and sequence 2 being at least substantially complementary, and each nucleotide of the sequence 1 and sequence 2 being a modified nucleotide;

[0577] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification;

[0578] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification;

[0579] When sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 7th, 8th, and 9th positions at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 14th, 17th, or 19th positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0580] When sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 8th, 9th, and 10th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 15th, 18th, or 20th positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0581] When sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 9th, 10th, and 11th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 16th, 19th, or 21st positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0582] Sequence 2: 1) The nucleotides at positions 2 and 14 of the 5' end contain a 2'-F modification; 2) The nucleotide at position 6 of the 5' end contains a 2'-OMe modification or a 2'-deoxy modification; 3) The nucleotide at position 16 contains a 2'-F modification, a 2'-OMe modification, or a 2'-deoxy modification; or,

[0583] Sequence 2: 1) The nucleotides at positions 2, 14, and 16 of the 5' end contain 2'-F modification, and 2) The nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification.

[0584] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, the sequence 1 and sequence 2 being at least substantially complementary, and each nucleotide of the sequence 1 and sequence 2 being a modified nucleotide;

[0585] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification;

[0586] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification;

[0587] When sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 7th, 8th, and 9th positions at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 14th, 17th, or 19th positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0588] When sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 8th, 9th, and 10th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 15th, 18th, or 20th positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0589] When sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 9th, 10th, and 11th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 16th, 19th, or 21st positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0590] Sequence 2: 1) The nucleotides at positions 2 and 14 of the 5' end contain a 2'-F modification; 2) The nucleotide at position 6 of the 5' end contains a 2'-OMe modification or a 2'-deoxy modification; 3) The nucleotide at position 16 contains a 2'-F modification, a 2'-OMe modification, or a 2'-deoxy modification; or,

[0591] Sequence 2: 1) The nucleotides at positions 2, 14, and 16 of the 5' end contain a 2'-F modification, and 2) The nucleotide at position 6 of the 5' end contains a 2'-OMe modification or a 2'-deoxy modification;

[0592] The remaining nucleotides of sequence 1 and sequence 2 contain 2'-OMe modification.

[0593] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, the sequence 1 and sequence 2 being at least substantially complementary, and each nucleotide of the sequence 1 and sequence 2 being a modified nucleotide;

[0594] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification;

[0595] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification;

[0596] When sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 7th, 8th, and 9th positions at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 14th, 17th, or 19th positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0597] When sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 8th, 9th, and 10th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 15th, 18th, or 20th positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0598] When sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 9th, 10th, and 11th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 16th, 19th, or 21st positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0599] When sequence 2 has 21 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, or 21 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-F modification.

[0600] When sequence 2 has 22 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, or 22 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-F modification.

[0601] When sequence 2 has 23 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-F modification.

[0602] or,

[0603] When sequence 2 has 21 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, or 21 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification.

[0604] When sequence 2 has 22 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, or 22 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification.

[0605] When sequence 2 has 23 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification.

[0606] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, the sequence 1 and sequence 2 being at least substantially complementary, and each nucleotide of the sequence 1 and sequence 2 being a modified nucleotide;

[0607] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification;

[0608] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification;

[0609] When sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 7th, 8th, and 9th positions at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 14th, 17th, or 19th positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0610] When sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 8th, 9th, and 10th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 15th, 18th, or 20th positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0611] When sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 9th, 10th, and 11th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 16th, 19th, or 21st positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0612] When sequence 2 has 21 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, or 21 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-F modification.

[0613] When sequence 2 has 22 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, or 22 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-F modification.

[0614] When sequence 2 has 23 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-F modification.

[0615] or,

[0616] When sequence 2 has 21 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, or 21 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification.

[0617] When sequence 2 has 22 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, or 22 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification.

[0618] When sequence 2 has 23 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 3) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end contains 2'-deoxy modification; 4) the nucleotide at position 6 of the 5' end contains 2'-OMe modification.

[0619] The remaining nucleotides of sequence 1 and sequence 2 contain 2'-OMe modification.

[0620] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, the sequence 1 and sequence 2 being at least substantially complementary, and each nucleotide of the sequence 1 and sequence 2 being a modified nucleotide;

[0621] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification;

[0622] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification;

[0623] When sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 7th, 8th, and 9th positions at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 14th, 17th, or 19th positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0624] When sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 8th, 9th, and 10th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 15th, 18th, or 20th positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0625] When sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 9th, 10th, and 11th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 16th, 19th, or 21st positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0626] When sequence 2 has 21 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification; 3) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, or 21 of the 5' end contains 2'-deoxy modification.

[0627] When sequence 2 has 22 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification; 3) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, or 22 of the 5' end contains 2'-deoxy modification.

[0628] When sequence 2 has 23 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification; 3) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end contains 2'-deoxy modification.

[0629] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 14 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising sequence 1, the antisense strand comprising sequence 2, the sequence 1 and sequence 2 being at least substantially complementary, and each nucleotide of the sequence 1 and sequence 2 being a modified nucleotide;

[0630] The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification;

[0631] The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification;

[0632] When sequence 1 has 19 nucleotides: 1) the 5th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 7th, 8th, and 9th positions at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 14th, 17th, or 19th positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0633] When sequence 1 has 20 nucleotides: 1) the 6th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 8th, 9th, and 10th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 15th, 18th, or 20th positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0634] When sequence 1 has 21 nucleotides: 1) the 7th nucleotide at the 5' end contains a 2'-OMe modification or a 2'-F modification; 2) two of the nucleotides at the 9th, 10th, and 11th nucleotides at the 5' end contain a 2'-F modification, and the remaining nucleotide contains a 2'-OMe modification or a 2'-deoxy modification; 3) one or more of the 16th, 19th, or 21st positions at the 5' end of sequence 1 independently contain a non-natural nucleotide, the base of which is selected from the following structure X:

[0635] When sequence 2 has 21 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification; 3) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, or 21 of the 5' end contains 2'-deoxy modification.

[0636] When sequence 2 has 22 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification; 3) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, or 22 of the 5' end contains 2'-deoxy modification.

[0637] When sequence 2 has 23 nucleotides: 1) the nucleotides at positions 2 and 14 of the 5' end contain 2'-F modification; 2) the nucleotide at position 6 of the 5' end contains 2'-OMe modification or 2'-deoxy modification; 3) the nucleotide at position 16 of the 5' end contains 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; 4) one of the nucleotides at positions 15, 16, 17, 18, 19, 20, 21, 22, or 23 of the 5' end contains 2'-deoxy modification.

[0638] The remaining nucleotides of sequence 1 and sequence 2 contain 2'-OMe modification.

[0639] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising the sequence shown in formula (I), and the antisense strand comprising the sequence shown in formula (II).

[0640] Formula (I): 5'-(Na) x N1N2N3N4N5N6N7N8N9N10N11N12N13N14N15N16N17N18N19-3',

[0641] Formula (II): 5'-N1'N2'N3'N4'N5'N6'N7'N8'N9'N10'N11'N12'N13'N14'N15'N16'N17'N18'N19'N20'N21'(Nb) y -3';

[0642] Each of Na, N1, N2, N3, N4, N6, N10, N11, N12, N13, N14, N15, N16, N17, N18, and N19 independently represents a modified nucleotide, the modification including a 2'-OMe modification.

[0643] The N5 represents a modified nucleotide, the modification comprising either a 2'-OMe modification or a 2'-F modification.

[0644] N7, N8, and N9 each independently represent a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification, wherein two of N7, N8, and N9 contain 2'-F modification, and each of the remaining ones contains either 2'-OMe modification or 2'-deoxy modification.

[0645] The x is selected from 0, 1, or 2; when x is selected from 1, Na represents a modified nucleotide Na1; when x is selected from 2, Na represents two modified nucleotides Na1Na2, wherein Na1 is linked to N1.

[0646] N1', N3', N4', N5', N7', N8', N9', N10', N11', N12', and N13' each independently represent a modified nucleotide, the modification including a 2'-OMe modification.

[0647] N2' and N14' each independently represent a modified nucleotide, the modification including a 2'-F modification.

[0648] The N6' represents a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification.

[0649] The N15', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification.

[0650] The N16' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification.

[0651] or,

[0652] N15', N17', N18', N19', and N20' each independently represent a modified nucleotide, the modification including a 2'-OMe modification.

[0653] Each of N21' and each Nb independently represents a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification.

[0654] The N16' represents a modified nucleotide, the modification including 2'-OMe modification or 2'-F modification.

[0655] The y is selected from 0, 1 or 2; when y is selected from 1, Nb represents a modified nucleotide Nb1; when y is selected from 2, Nb represents two modified nucleotides Nb1Nb2, wherein Nb1 is connected to N21'.

[0656] Each of N14, N17, or N19 independently comprises one or more non-natural nucleotide bases, the bases of which are selected from the following structure X:

[0657] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising the sequence shown in formula (I), and the antisense strand comprising the sequence shown in formula (II).

[0658] Formula (I): 5'-(Na)xN1N2N3N4N5N6N7N8N9N10N11N12N13N14N15N16N17N18N19-3',

[0659] Formula (II): 5'-N1'N2'N3'N4'N5'N6'N7'N8'N9'N10'N11'N12'N13'N14'N15'N16'N17'N18'N19'N20'N21'(Nb)y-3';

[0660] Each of Na, N1, N2, N3, N4, N6, N10, N11, N12, N13, N14, N15, N16, N17, N18, and N19 independently represents a modified nucleotide, the modification including a 2'-OMe modification.

[0661] The N5 represents a modified nucleotide, the modification comprising either a 2'-OMe modification or a 2'-F modification.

[0662] N7, N8, and N9 each independently represent a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification, wherein two of N7, N8, and N9 contain 2'-F, and each of the remaining ones contains either 2'-OMe modification or 2'-deoxy modification.

[0663] The x is selected from 0, 1, or 2; when x is selected from 1, Na represents a modified nucleotide Na1; when x is selected from 2, Na represents two modified nucleotides Na1Na2, wherein Na1 is linked to N1.

[0664] N1', N3', N4', N5', N7', N8', N9', N10', N11', N12', and N13' each independently represent a modified nucleotide, the modification including a 2'-OMe modification.

[0665] N2' and N14' each independently represent a modified nucleotide, the modification including a 2'-F modification.

[0666] The N6' represents a modified nucleotide, the modification including 2'-F modification or 2'-OMe modification.

[0667] N15', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification; N16' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; wherein, a nucleotide includes 2'-deoxy modification, or,

[0668] Each of N15', N16', N17', N18', N19', N20', N21' and each Nb independently represents a modified nucleotide, wherein one of the nucleotides contains a 2'-deoxy modification. Except for the nucleotides containing a 2'-deoxy modification, each of the remaining N15', N17', N18', N19', N20', N21' and each Nb independently contains a 2'-OMe modification, and the remaining N16' contains either a 2'-OMe modification or a 2'-F modification.

[0669] The y is selected from 0, 1 or 2; when y is selected from 1, Nb represents a modified nucleotide Nb1; when y is selected from 2, Nb represents two modified nucleotides Nb1Nb2, wherein Nb1 is connected to N21'.

[0670] Each of N14, N17, or N19 independently comprises one or more non-natural nucleotide bases, the bases of which are selected from the following structure X:

[0671] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising the sequence shown in formula (I), and the antisense strand comprising the sequence shown in formula (II).

[0672] Formula (I): 5'-(Na)xN1N2N3N4N5N6N7N8N9N10N11N12N13N14N15N16N17N18N19-3',

[0673] Formula (II): 5'-N1'N2'N3'N4'N5'N6'N7'N8'N9'N10'N11'N12'N13'N14'N15'N16'N17'N18'N19'N20'N21'(Nb)y-3';

[0674] Each of Na, N1, N2, N3, N4, N6, N10, N11, N12, N13, N14, N15, N16, N17, N18, and N19 independently represents a modified nucleotide, the modification including a 2'-OMe modification.

[0675] The N5 represents a modified nucleotide, the modification comprising either a 2'-OMe modification or a 2'-F modification.

[0676] N7, N8, and N9 each independently represent a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification, wherein two of N7, N8, and N9 contain 2'-F, and each of the remaining ones contains either 2'-OMe modification or 2'-deoxy modification.

[0677] The x is selected from 0, 1, or 2; when x is selected from 1, Na represents a modified nucleotide Na1; when x is selected from 2, Na represents two modified nucleotides Na1Na2, wherein Na1 is linked to N1.

[0678] N1', N3', N4', N5', N7', N8', N9', N10', N11', N12', and N13' each independently represent a modified nucleotide, the modification including a 2'-OMe modification.

[0679] N2' and N14' each independently represent a modified nucleotide, the modification including a 2'-F modification.

[0680] The N6' represents a modified nucleotide, the modification comprising a 2'-OMe modification or a 2'-deoxy modification.

[0681] N15', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification; N16' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; wherein, a nucleotide includes 2'-deoxy modification, or,

[0682] Each of N15', N16', N17', N18', N19', N20', N21' and each Nb independently represents a modified nucleotide, wherein one of the nucleotides contains a 2'-deoxy modification. Except for the nucleotides containing a 2'-deoxy modification, each of the remaining N15', N17', N18', N19', N20', N21' and each Nb independently contains a 2'-OMe modification, and the remaining N16' contains either a 2'-OMe modification or a 2'-F modification.

[0683] The y is selected from 0, 1 or 2; when y is selected from 1, Nb represents a modified nucleotide Nb1; when y is selected from 2, Nb represents two modified nucleotides Nb1Nb2, wherein Nb1 is connected to N21'.

[0684] Each of N14, N17, or N19 independently comprises one or more non-natural nucleotide bases, the bases of which are selected from the following structure X:

[0685] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising the sequence shown in formula (I), the antisense strand comprising the sequence shown in formula (II), wherein n13, n14, n16, n17, or n19 each independently comprises one or more non-natural nucleotide bases, the bases of which are selected from the following structure X: Equations (I) and (II) are selected from the following groups in Table 7:

[0686] Table 7

[0687] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising the sequence shown in formula (I), the antisense strand comprising the sequence shown in formula (II), wherein n13, n14, n16, n17, or n19 each independently comprises one or more non-natural nucleotide bases, the bases of which are selected from the following structure X: Equations (I) and (II) are selected from the following groups in Table 8:

[0688] Table 8

[0689] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising the sequence shown in formula (I), the antisense strand comprising the sequence shown in formula (II), wherein n13, n14, n16, n17, or n19 each independently comprises one or more non-natural nucleotide bases, the bases of which are selected from the following structure X: Equations (I) and (II) are selected from the following groups in Table 9:

[0690] Table 9

[0691] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, each strand having 19 to 40 nucleotides, the antisense strand being at least substantially complementary to a target gene, the sense strand comprising the sequence shown in formula (I), the antisense strand comprising the sequence shown in formula (II), wherein n13, n14, n16, n17, or n19 each independently comprises one or more non-natural nucleotide bases, the bases of which are selected from the following structure X: Equations (I) and (II) are selected from the following groups in Table 10:

[0692] Table 10

[0693] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, the antisense strand being at least substantially complementary to a target gene, and each of n13, n14, n16, n17, or n19 independently comprises one or more non-natural nucleotide bases, the bases of which are selected from the following structure X: The justice chain or antisense chain is selected from the following groups in Table 11:

[0694] Table 11

[0695] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the dsRNA comprises a sense strand and an antisense strand, the antisense strand being at least substantially complementary to a target gene, and each of n13, n14, n16, n17, or n19 independently comprises one or more non-natural nucleotide bases, the bases of which are selected from the following structure X: The justice chain or antisense chain is selected from the following groups in Table 12:

[0696] Table 12

[0697] Targeting PNPLA3 mRNA

[0698] This application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the sense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.1, and the antisense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.2, and the sense or antisense strand has 19-23 nucleotides (e.g., 19, 20, 21, 22, or 23 nucleotides):

[0699] 5'-CUUGCUACCCAUUAGGAUX 1 -3'(SEQ ID NO.1),

[0700] 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 -3'(SEQ ID NO.2);

[0701] The X 1 Y 1 Y 2 Or Y 3 Selected from A, U, C, T, or G;

[0702] The justice chain has the sequence shown in equation (I) above, where x is 0, 1, or 2.

[0703] The antisense chain has the sequence shown in equation (II) above, where y is 0, 1 or 2.

[0704] This application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the sense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.1, with a difference of no more than 3 nucleotides, and the antisense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.2, with a difference of no more than 3 nucleotides.

[0705] 5'-CUUGCUACCCAUUAGGAUX 1 -3'(SEQ ID NO.1),

[0706] 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 -3'(SEQ ID NO.2);

[0707] The X 1 Y 1 Y 2 Or Y 3 Each is independently selected from A, U, C, or G.

[0708] The justice chain or antisense chain is optionally modified.

[0709] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the sense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.1, differing by no more than 3 nucleotides:

[0710] 5'-CUUGCUACCCAUUAGGAUX 1 -3'(SEQ ID NO.1);

[0711] The X 1 Choose from A, U, C or G.

[0712] The chain of justice may optionally be modified.

[0713] In some embodiments, the sense strand of the dsRNA targeting PNPLA3 mRNA has the sequence shown in formula (I) above, where x is 0.

[0714] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the antisense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.2, differing by no more than 3 nucleotides:

[0715] 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 -3'(SEQ ID NO.2);

[0716] The Y 1 Y 2 Or Y 3 Each is independently selected from A, U, C, T, or G.

[0717] The antisense chain is optionally modified.

[0718] In some embodiments, the antisense strand of the dsRNA targeting PNPLA3 mRNA has the sequence shown in formula (II) above, where y is 0.

[0719] This application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the sense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.1, with a difference of no more than 3 nucleotides, and the antisense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.2, with a difference of no more than 3 nucleotides.

[0720] 5'-CUUGCUACCCAUUAGGAUX 1 -3'(SEQ ID NO.1),

[0721] 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 -3'(SEQ ID NO.2);

[0722] The X 1 Y 1 Y 2 Or Y 3 Each is independently selected from A, U, C, T, or G.

[0723] The positive chain comprises a sequence in which one or more non-natural nucleotides are replaced by non-natural nucleotides in the sequence shown in SEQ ID NO.1.

[0724] The antisense strand comprises a sequence in which one or more non-natural nucleotides are replaced by non-natural nucleotides in the sequence shown in SEQ ID NO.2.

[0725] The justice chain or antisense chain is optionally modified.

[0726] This application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the sense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.1, with a difference of no more than 3 nucleotides, and the antisense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.2, with a difference of no more than 3 nucleotides.

[0727] 5'-CUUGCUACCCAUUAGGAUX 1 -3'(SEQ ID NO.1),

[0728] 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 -3'(SEQ ID NO.2);

[0729] The X 1 Y 1 Y 2 Or Y 3 Each is independently selected from A, U, C, or G.

[0730] The justice chain has the sequence shown in equation (I) above, where x is 0, 1, or 2, preferably 0.

[0731] The antisense chain has the sequence shown in formula (II) above, where y is 0, 1, or 2, preferably 0.

[0732] The positive chain comprises a sequence in which one or more non-natural nucleotides are replaced by non-natural nucleotides in the sequence shown in SEQ ID NO.1.

[0733] The antisense strand comprises a sequence in which one or more non-natural base nucleotides are replaced with non-natural base nucleotides in the sequence shown in SEQ ID NO.2.

[0734] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the sense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.1, differing by no more than 3 nucleotides:

[0735] 5'-CUUGCUACCCAUUAGGAUX 1 -3'(SEQ ID NO.1);

[0736] The X 1 Choose from A, U, C or G.

[0737] The positive chain comprises a sequence in which one or more non-natural nucleotides are replaced by non-natural nucleotides in the sequence shown in SEQ ID NO.1.

[0738] The chain of justice may optionally be modified.

[0739] This application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the antisense strand contains at least 15 consecutive nucleotides in the nucleotide sequence shown in SEQ ID NO.2, with a difference of no more than 3 nucleotides:

[0740] 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 -3'(SEQ ID NO.2);

[0741] The Y 1 Y 2 Or Y 3 Each is independently selected from A, U, C, or G.

[0742] The antisense strand comprises a sequence in which one or more non-natural nucleotides are replaced by non-natural nucleotides in the sequence shown in SEQ ID NO.2.

[0743] The antisense chain is optionally modified.

[0744] This application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the sense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.1, with a difference of no more than 3 nucleotides, and the antisense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.2, with a difference of no more than 3 nucleotides.

[0745] 5'-CUUGCUACCCAUUAGGAUX 1 -3'(SEQ ID NO.1),

[0746] 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 -3'(SEQ ID NO.2);

[0747] The X 1 Y 1 Y 2 Or Y 3 Each is independently selected from A, U, C, or G.

[0748] The nucleotides at positions 1, 2, 3, 4, 6, 10, 11, 12, 13, 14, 15, 16, 17, 18, and 19 of the 5' end of the positive strand are either unmodified or modified nucleotides.

[0749] The 5th nucleotide at the 5' end of the positive strand contains a 2'-OMe modification or a 2'-F modification.

[0750] The nucleotides at positions 7, 8, and 9 of the 5' end of the positive strand contain 2'-OMe, 2'-F, or 2'-deoxy modifications, and are not all selected from 2'-F modifications;

[0751] The nucleotides at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, and 13 of the 5' end of the antisense strand are either unmodified or modified nucleotides.

[0752] The nucleotides at positions 2 and 14 of the 5' end of the antisense strand are modified nucleotides.

[0753] The nucleotide at position 6 of the 5' end of the antisense strand contains a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification.

[0754] The nucleotides at positions 15, 17, 18, 19, 20, and 21 of the 5' end of the antisense strand contain a 2'-OMe modification or a 2'-deoxy modification.

[0755] The nucleotide at position 16 of the 5' end of the antisense strand contains a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification.

[0756] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the sense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.1, differing by no more than 3 nucleotides:

[0757] 5'-CUUGCUACCCAUUAGGAUX 1 -3'(SEQ ID NO.1);

[0758] The nucleotides at positions 1, 2, 3, 4, 6, 10, 11, 12, 13, 14, 15, 16, 17, 18, and 19 of the 5' end of the positive strand are either unmodified or modified nucleotides.

[0759] The X 1 Choose from A, U, C or G.

[0760] The 5th nucleotide at the 5' end of the positive strand contains a 2'-OMe modification or a 2'-F modification.

[0761] The nucleotides at positions 7, 8, and 9 of the 5' end of the positive strand contain 2'-OMe, 2'-F, or 2'-deoxy modifications, and are not all selected from 2'-F modifications.

[0762] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the antisense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.2, differing by no more than 3 nucleotides:

[0763] 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 -3'(SEQ ID NO.2);

[0764] The Y 1 Y 2 Or Y 3 Each is independently selected from A, U, C, or G.

[0765] The nucleotides at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, and 13 of the 5' end of the antisense strand are either unmodified or modified nucleotides.

[0766] The nucleotides at positions 2 and 14 of the 5' end of the antisense strand are modified nucleotides.

[0767] The nucleotide at position 6 of the 5' end of the antisense strand contains a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification.

[0768] The nucleotides at positions 15, 17, 18, 19, 20, and 21 of the 5' end of the antisense strand contain a 2'-OMe modification or a 2'-deoxy modification.

[0769] The nucleotide at position 16 of the 5' end of the antisense strand contains a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification.

[0770] In some embodiments, the X 1 Selected from A or U. In some embodiments, X 1 Selected from A.

[0771] In some embodiments, the Y 1 Selected from A or U. In some embodiments, Y 1 Selected from U.

[0772] In some embodiments, the Y 2 Selected from A, U, C, or G. In some embodiments, the Y... 2 Selected from U.

[0773] In some embodiments, the Y 3 Selected from U, C, or G. In some embodiments, the Y 3 Selected from U or T.

[0774] This application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the antisense strand is at least substantially complementary to the target gene, the sense strand comprises sequence 1, the antisense strand comprises sequence 2, and sequences 1 and 2 are at least substantially complementary. Sequence 1 has the nucleotide sequence shown in SEQ ID NO. 1, and sequence 2 has the nucleotide sequence shown in SEQ ID NO. 1.

[0775] 5'-CUUGCUACCCAUUAGGAUX 1 -3'(SEQ ID NO.1),

[0776] 5'-Y 1 AUCCUAAUGGGUAGCAAGY2 Y 3 -3'(SEQ ID NO.2);

[0777] The X 1 Y 1 Y 2 Or Y 3 Each is independently selected from A, U, C, or G.

[0778] Optionally, the sense strand comprises a sequence in which one or more non-natural nucleotides are replaced by non-natural nucleotides in the sequence shown in SEQ ID NO.1, or the antisense strand comprises a sequence in which one or more non-natural nucleotides are replaced by non-natural nucleotides in the sequence shown in SEQ ID NO.2.

[0779] The modifications for Sequence 1 and Sequence 2 are defined as above.

[0780] In some embodiments, the positive chain comprises a sequence in which one or more non-natural base nucleotides are replaced with non-natural base nucleotides in a sequence as shown in SEQ ID NO.1.

[0781] In some embodiments, the positive chain includes a sequence in which one, two, three, four, or five non-natural nucleotides are replaced with non-natural nucleotides in a sequence as shown in SEQ ID NO.1.

[0782] In some embodiments, the positive chain comprises a sequence in which one or two nucleotides are replaced with non-natural base nucleotides in a sequence as shown in SEQ ID NO.1.

[0783] In some embodiments, the positive chain comprises a sequence in which a non-natural base nucleotide is replaced with a nucleotide in the sequence shown in SEQ ID NO.1.

[0784] In some embodiments, the antisense strand comprises a sequence in which one or more nucleotides are replaced with non-natural base nucleotides in a sequence as shown in SEQ ID NO.2.

[0785] In some embodiments, the antisense strand comprises a sequence in which one, two, three, four, or five nucleotides are replaced with non-natural base nucleotides in a sequence as shown in SEQ ID NO.2.

[0786] In some embodiments, the antisense strand comprises a sequence in which one or two nucleotides are replaced with non-natural base nucleotides in a sequence as shown in SEQ ID NO.2.

[0787] In some embodiments, the antisense strand comprises a sequence in which a non-natural base nucleotide is replaced with a nucleotide in a sequence as shown in SEQ ID NO.2.

[0788] In some embodiments, the positive strand includes a sequence in which one or more nucleotides are replaced with non-natural base nucleotides in the sequence shown in SEQ ID NO.1, wherein the non-natural base nucleotide substitutions occur at any position in SEQ ID NO.1.

[0789] In some embodiments, the positive strand includes a sequence in which one or more nucleotides are replaced by non-natural base nucleotides in the sequence shown in SEQ ID NO.1, wherein the non-natural base nucleotide replacement occurs at positions 1 to 19 of the 5' end of SEQ ID NO.1.

[0790] In some embodiments, the positive strand includes a sequence in which one or more nucleotides are replaced by non-natural base nucleotides in the sequence shown in SEQ ID NO.1, wherein the non-natural base nucleotide substitution occurs at position 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 or 19 at the 5' end of SEQ ID NO.1.

[0791] In some embodiments, the positive strand includes a sequence in which one or more nucleotides are replaced by non-natural base nucleotides in the sequence shown in SEQ ID NO.1, wherein the non-natural base nucleotide substitutions occur at position 12, 13, 14, 15, 16, 17, 18 or 19 at the 5' end of SEQ ID NO.1.

[0792] In some embodiments, the positive strand includes a sequence in which one or more nucleotides are replaced by non-natural base nucleotides in the sequence shown in SEQ ID NO.1, wherein the non-natural base nucleotide substitution occurs at position 14, 17 or 19 of the 5' end of SEQ ID NO.1.

[0793] In some embodiments, the antisense strand comprises a sequence in which one or more nucleotides are replaced by non-natural base nucleotides in the sequence shown in SEQ ID NO.2, wherein the non-natural base nucleotide substitutions occur at any position in SEQ ID NO.2.

[0794] In some embodiments, the antisense strand comprises a sequence in which one or more nucleotides are replaced by non-natural base nucleotides in the sequence shown in SEQ ID NO.2, wherein the non-natural base nucleotide replacement occurs at positions 1 to 21 of the 5' end of SEQ ID NO.2.

[0795] In some embodiments, the antisense strand comprises a sequence in which one or more nucleotides are replaced by non-natural base nucleotides in the sequence shown in SEQ ID NO.2, wherein the non-natural base nucleotide substitution occurs at position 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 at the 5' end of SEQ ID NO.2.

[0796] In some embodiments, the bases of the non-natural nucleotide are selected from the following structure X:

[0797] In some embodiments, the non-natural nucleoside is selected from (x):

[0798] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 1, 2, 3, 4, 6, 10, 11, 12, 13, 14, 15, 16, 17, 18 and 19 of the 5' end of the -3'(SEQ ID NO.1) are modified nucleotides, and the modifications do not include 2'-F modification.

[0799] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 1, 2, 3, 4, 6, 10, 11, 12, 13, 14, 15, 16, 17, 18 and 19 of the 5' end of the -3'(SEQ ID NO.1) are modified nucleotides, including 2'-OMe modification.

[0800] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotide at the 5th position of the 5' end of the -3' (SEQ ID NO.1) contains a 2'-F modification.

[0801] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The 5th nucleotide at the 5' end of the -3'(SEQ ID NO.1) contains a 2'-OMe modification.

[0802] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 7, 8 and 9 of the 5' end of the -3' (SEQ ID NO.1) contain a 2'-OMe modification, a 2'-F modification or a 2'-deoxy modification, and at least one (e.g. one or two) nucleotides contain a 2'-F modification.

[0803] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 7, 8 and 9 of the 5' end of the -3'(SEQ ID NO.1) contain 2'-OMe modification, 2'-F modification or 2'-deoxy modification, and are not all selected from 2'-OMe modification or 2'-F modification.

[0804] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 One of the nucleotides at positions 7, 8 and 9 of the 5' end of the -3'(SEQ ID NO.1) contains a 2'-F modification, and each of the remaining nucleotides independently contains a 2'-OMe modification or a 2'-deoxy modification.

[0805] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The 7th nucleotide at the 5' end of the -3' (SEQ ID NO.1) contains a 2'-F modification, and the 8th and 9th nucleotides each independently contain a 2'-OMe modification or a 2'-deoxy modification.

[0806] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The 7th nucleotide at the 5' end of the -3'(SEQ ID NO.1) contains a 2'-F modification, the 8th nucleotide contains a 2'-OMe modification, and the 9th nucleotide contains a 2'-OMe modification.

[0807] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotide at position 7 of the 5' end of the -3'(SEQ ID NO.1) contains a 2'-F modification, the nucleotide at position 8 contains a 2'-deoxy modification, and the nucleotide at position 9 contains a 2'-deoxy modification.

[0808] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1The 7th nucleotide at the 5' end of the -3'(SEQ ID NO.1) contains a 2'-F modification, the 8th nucleotide contains a 2'-OMe modification, and the 9th nucleotide contains a 2'-deoxy modification.

[0809] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The 7th nucleotide at the 5' end of the -3'(SEQ ID NO.1) contains a 2'-F modification, the 8th nucleotide contains a 2'-deoxy modification, and the 9th nucleotide contains a 2'-OMe modification.

[0810] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotide at position 8 of the 5' end of the -3' (SEQ ID NO.1) contains a 2'-F modification, and the nucleotides at positions 7 and 9 each contain either a 2'-OMe modification or a 2'-deoxy modification.

[0811] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotide at position 8 of the 5' end of the -3'(SEQ ID NO.1) contains a 2'-F modification, the nucleotide at position 7 contains a 2'-OMe modification, and the nucleotide at position 9 contains a 2'-OMe modification.

[0812] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotide at position 8 of the 5' end of the -3'(SEQ ID NO.1) contains a 2'-F modification, the nucleotide at position 7 contains a 2'-deoxy modification, and the nucleotide at position 9 contains a 2'-deoxy modification.

[0813] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The 8th nucleotide at the 5' end of the -3'(SEQ ID NO.1) contains a 2'-F modification, the 7th nucleotide contains a 2'-OMe modification, and the 9th nucleotide contains a 2'-deoxy modification.

[0814] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotide at position 8 of the 5' end of the -3'(SEQ ID NO.1) contains a 2'-F modification, the nucleotide at position 7 contains a 2'-deoxy modification, and the nucleotide at position 9 contains a 2'-OMe modification.

[0815] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1The nucleotide at position 9 of the 5' end of the -3' (SEQ ID NO.1) contains a 2'-F modification, and the nucleotides at positions 7 and 8 each contain either a 2'-OMe modification or a 2'-deoxy modification.

[0816] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotide at position 9 of the 5' end of the -3'(SEQ ID NO.1) contains a 2'-F modification, the nucleotide at position 7 contains a 2'-OMe modification, and the nucleotide at position 8 contains a 2'-OMe modification.

[0817] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotide at position 9 of the 5' end of the -3'(SEQ ID NO.1) contains a 2'-F modification, the nucleotide at position 7 contains a 2'-deoxy modification, and the nucleotide at position 8 contains a 2'-deoxy modification.

[0818] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotide at position 9 of the 5' end of the -3'(SEQ ID NO.1) contains a 2'-F modification, the nucleotide at position 7 contains a 2'-OMe modification, and the nucleotide at position 8 contains a 2'-deoxy modification.

[0819] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotide at position 9 of the 5' end of the -3'(SEQ ID NO.1) contains a 2'-F modification, the nucleotide at position 7 contains a 2'-deoxy modification, and the nucleotide at position 8 contains a 2'-OMe modification.

[0820] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 Two of the nucleotides at positions 7, 8, and 9 of the 5' end of the -3'(SEQ ID NO.1) contain a 2'-F modification, and each of the remaining nucleotides independently contains either a 2'-OMe modification or a 2'-deoxy modification.

[0821] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 7 and 8 of the 5' end of the -3' (SEQ ID NO.1) contain a 2'-F modification, and the nucleotide at position 9 contains a 2'-OMe modification or a 2'-deoxy modification.

[0822] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1The nucleotides at positions 7 and 8 of the 5' end of the -3'(SEQ ID NO.1) contain a 2'-F modification, and the nucleotide at position 9 contains a 2'-OMe modification.

[0823] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 7 and 8 of the 5' end of the -3' (SEQ ID NO.1) contain a 2'-F modification, and the nucleotide at position 9 contains a 2'-deoxy modification.

[0824] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 7 and 9 of the 5' end of the -3' (SEQ ID NO.1) contain a 2'-F modification, and the nucleotide at position 8 contains a 2'-OMe modification or a 2'-deoxy modification.

[0825] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 7 and 9 of the 5' end of the -3' (SEQ ID NO.1) contain a 2'-F modification, and the nucleotide at position 8 contains a 2'-OMe modification.

[0826] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 7 and 9 of the 5' end of the -3' (SEQ ID NO.1) contain a 2'-F modification, and the nucleotide at position 8 contains a 2'-deoxy modification.

[0827] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 8 and 9 of the 5' end of the -3' (SEQ ID NO.1) contain a 2'-F modification, and the nucleotide at position 7 contains a 2'-OMe modification or a 2'-deoxy modification.

[0828] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 8 and 9 of the 5' end of the -3' (SEQ ID NO.1) contain a 2'-F modification, and the nucleotide at position 7 contains a 2'-OMe modification.

[0829] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 8 and 9 of the 5' end of the -3' (SEQ ID NO.1) contain a 2'-F modification, and the nucleotide at position 7 contains a 2'-deoxy modification.

[0830] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, and 13 of the 5' end of the -3'(SEQ ID NO.2) are modified nucleotides, excluding the 2'-F modification.

[0831] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, and 13 of the 5' end of the -3'(SEQ ID NO.2) are modified nucleotides, and the modification includes 2'-OMe modification.

[0832] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 -3'(SEQ ID NO.2) The nucleotides at positions 2 and 14 of the 5' end are modified nucleotides, and the modification does not include 2'-OMe modification.

[0833] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 -3'(SEQ ID NO.2) The nucleotides at positions 2 and 14 of the 5' end are modified nucleotides, and the modification includes 2'-F modification.

[0834] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotide at position 6 of the 5' end of the -3' (SEQ ID NO.2) contains a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification.

[0835] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The 6th nucleotide at the 5' end of the -3' (SEQ ID NO.2) contains a 2'-F modification.

[0836] In some embodiments, the antisense chain 5'-Y 1AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotide at position 6 of the 5' end of the -3' (SEQ ID NO.2) contains a 2'-OMe modification or a 2'-deoxy modification.

[0837] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) do not contain 2'-deoxy modification.

[0838] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 At least one of the nucleotides at positions 15, 16, 17, 18, 19, 20 and 21 of the 5' end of the -3'(SEQ ID NO.2) contains a 2'-deoxy modification.

[0839] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 Three, two, or one of the nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) contains a 2'-deoxy modification.

[0840] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 One of the nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) contains a 2'-deoxy modification.

[0841] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20 and 21 of the 5' end of the -3'(SEQ ID NO.2) contain 2'-OMe modification or 2'-deoxy modification, and the nucleotide at position 16 contains 2'-OMe modification, 2'-F modification or 2'-deoxy modification. Two of the above nucleotides contain 2'-deoxy modification.

[0842] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20 and 21 of the 5' end of the -3'(SEQ ID NO.2) contain a 2'-OMe modification or a 2'-deoxy modification, and the nucleotide at position 16 contains a 2'-OMe modification, a 2'-F modification or a 2'-deoxy modification. One of the above nucleotides contains a 2'-deoxy modification.

[0843] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 Two nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) contain a 2'-deoxy modification, the remaining nucleotides at positions 15, 17, 18, 19, 20, or 21 contain a 2'-OMe modification, and the remaining nucleotide at position 16 contains either a 2'-OMe modification or a 2'-F modification.

[0844] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 One of the nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) contains a 2'-deoxy modification, the remaining nucleotides at positions 15, 17, 18, 19, 20, or 21 contain a 2'-OMe modification, and the remaining nucleotide at position 16 contains either a 2'-OMe modification or a 2'-F modification.

[0845] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) each independently represent a modified nucleotide, with only position 15 containing a 2'-deoxy modification.

[0846] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) each independently represent a modified nucleotide, with only position 15 containing a 2'-deoxy modification, position 16 containing a 2'-F modification, and positions 17, 18, 19, 20, and 21 containing a 2'-OMe modification.

[0847] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) each independently represent a modified nucleotide, of which only the nucleotide at position 16 contains either a 2'-OMe modification or a 2'-deoxy modification.

[0848] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3' (SEQ ID NO.2) each independently represent a modified nucleotide, of which only nucleotide 16 contains a 2'-OMe modification or a 2'-deoxy modification, and nucleotides 15, 17, 18, 19, 20, and 21 contain a 2'-OMe modification.

[0849] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) each independently represent a modified nucleotide, with only the nucleotide at position 16 containing a 2'-deoxy modification.

[0850] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) each independently represent a modified nucleotide, of which only position 16 contains a 2'-deoxy modification, and positions 15, 17, 18, 19, 20, and 21 contain a 2'-OMe modification.

[0851] In some embodiments, the antisense chain 5'-Y 1AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3' (SEQ ID NO.2) each independently represent a modified nucleotide containing the 2'-OMe modification.

[0852] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) each independently represent a modified nucleotide, with only position 17 containing a 2'-deoxy modification.

[0853] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) each independently represent a modified nucleotide, with only position 17 containing a 2'-deoxy modification, position 16 containing a 2'-F modification, and positions 15, 18, 19, 20, and 21 containing a 2'-OMe modification.

[0854] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) each independently represent a modified nucleotide, with only position 18 containing a 2'-deoxy modification.

[0855] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) each independently represent a modified nucleotide, with only position 18 containing a 2'-deoxy modification, position 16 containing a 2'-F modification, and positions 15, 17, 19, 20, and 21 containing a 2'-OMe modification.

[0856] In some embodiments, the antisense chain 5'-Y 1AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3' (SEQ ID NO.2) each independently represent a modified nucleotide, with only nucleotide position 19 containing a 2'-deoxy modification.

[0857] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) each independently represent a modified nucleotide, with only nucleotide 19 containing a 2'-deoxy modification, nucleotide 16 containing a 2'-F modification, and nucleotides 15, 17, 18, 20, and 21 containing a 2'-OMe modification.

[0858] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3' (SEQ ID NO.2) each independently represent a modified nucleotide, with only the nucleotide at position 20 containing a 2'-deoxy modification.

[0859] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) each independently represent a modified nucleotide, with only position 20 containing a 2'-deoxy modification, position 16 containing a 2'-F modification, and positions 15, 17, 18, 19, and 21 containing a 2'-OMe modification.

[0860] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20 and 21 of the 5' end of the -3'(SEQ ID NO.2) each independently represent a modified nucleotide, of which only the nucleotide at position 21 contains a 2'-deoxy modification.

[0861] In some embodiments, the antisense chain 5'-Y1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the -3'(SEQ ID NO.2) each independently represent a modified nucleotide, with only position 21 containing a 2'-deoxy modification, position 16 containing a 2'-F modification, and positions 15, 17, 18, 19, and 20 containing a 2'-OMe modification.

[0862] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotide at position 16 of the 5' end of the -3'(SEQ ID NO.2) contains a 2'-OMe modification.

[0863] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotide at position 16 of the 5' end of the -3'(SEQ ID NO.2) contains a 2'-F modification.

[0864] In some embodiments, the antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotide at position 16 of the 5' end of the -3'(SEQ ID NO.2) contains a 2'-deoxy modification.

[0865] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 1, 2, 3, 4, 6, 10, 11, 12, 13, 14, 15, 16, 17, 18, and 19 of the 5' end of the -3' (SEQ ID NO.1) each independently contain a 2'-OMe modification.

[0866] The 5th nucleotide at the 5' end of the positive strand contains a 2'-OMe modification or a 2'-F modification.

[0867] Two of the nucleotides at positions 7, 8, and 9 of the 5' end of the positive strand contain a 2'-F modification, and the remaining one contains a 2'-OMe modification or a 2'-deoxy modification;

[0868] The antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3The nucleotides at positions 2 and 14 of the 5' end of the -3' (SEQ ID NO.2) each independently contain a 2'-F modification.

[0869] The nucleotide at the 5' end of the antisense strand contains a 2'-OMe modification or a 2'-deoxy modification.

[0870] The nucleotides at positions 15, 17, 18, 19, 20, and 21 of the 5' end of the antisense strand each independently contain either a 2'-OMe modification or a 2'-deoxy modification.

[0871] The nucleotide at position 16 of the 5' end of the antisense strand contains a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification;

[0872] or,

[0873] The nucleotides at positions 15, 17, 18, 19, 20, and 21 of the 5' end of the antisense strand each independently contain a 2'-OMe modification.

[0874] The nucleotide at position 16 of the 5' end of the antisense strand contains a 2'-OMe modification or a 2'-F modification.

[0875] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 1, 2, 3, 4, 6, 10, 11, 12, 13, 14, 15, 16, 17, 18, and 19 of the 5' end of the -3' (SEQ ID NO.1) each independently contain a 2'-OMe modification.

[0876] The 5th nucleotide at the 5' end of the positive strand contains a 2'-OMe modification or a 2'-F modification.

[0877] Two of the nucleotides at positions 7, 8, and 9 of the 5' end of the positive strand contain a 2'-F modification, and the remaining one contains either a 2'-OMe modification or a 2'-deoxy modification.

[0878] The antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 2 and 14 of the 5' end of the -3' (SEQ ID NO.2) each independently contain a 2'-F modification.

[0879] The nucleotide at the 5' end of the antisense strand contains a 2'-F modification or a 2'-OMe modification;

[0880] The antisense strand has nucleotides at positions 15, 17, 18, 19, 20, and 21 at the 5' end modified with 2'-OMe or 2'-deoxy, and nucleotide at position 16 contains 2'-OMe, 2'-F, or 2'-deoxy modification; wherein, one of the nucleotides contains 2'-deoxy modification.

[0881] or,

[0882] The nucleotide at the 5' end of the antisense strand contains a 2'-F modification.

[0883] One of the nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the antisense strand contains a 2'-deoxy modification. Apart from the nucleotide containing the 2'-deoxy modification, each of the remaining nucleotides at positions 15, 17, 18, 19, 20, and 21 of the 5' end of the antisense strand independently contains a 2'-OMe modification. The remaining nucleotide at position 16 of the 5' end of the antisense strand contains either a 2'-OMe modification or a 2'-F modification.

[0884] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 1, 2, 3, 4, 6, 10, 11, 12, 13, 14, 15, 16, 17, 18, and 19 of the 5' end of the -3' (SEQ ID NO.1) each independently contain a 2'-OMe modification.

[0885] The 5th nucleotide at the 5' end of the positive strand contains a 2'-OMe modification or a 2'-F modification.

[0886] Two of the nucleotides at positions 7, 8, and 9 of the 5' end of the positive strand contain a 2'-F modification, and the remaining one contains either a 2'-OMe modification or a 2'-deoxy modification.

[0887] The antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 2 and 14 of the 5' end of the -3' (SEQ ID NO.2) each independently contain a 2'-F modification.

[0888] The nucleotide at the 5' end of the antisense strand contains a 2'-OMe modification or a 2'-deoxy modification.

[0889] The antisense strand has nucleotides at positions 15, 17, 18, 19, 20, and 21 at the 5' end modified with 2'-OMe or 2'-deoxy, and nucleotide at position 16 contains 2'-OMe, 2'-F, or 2'-deoxy modification; wherein, one of the nucleotides contains 2'-deoxy modification.

[0890] or

[0891] One of the nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 at the 5'-end of the antisense strand contains a 2'-deoxy modification. Except for the nucleotide containing the 2'-deoxy modification, each of the remaining nucleotides at positions 15, 17, 18, 19, 20, and 21 at the 5'-end of the antisense strand independently contains a 2'-OMe modification, and the remaining nucleotide at position 16 at the 5'-end of the antisense strand contains a 2'-OMe modification or a 2'-F modification.

[0892] In some embodiments, the sense strand is 5'-CUUGCUACCCAUUAGGAUX 1 -3'(SEQ ID NO.1), where X 1 is selected from A or U, preferably A, and the sense strand comprises a sequence in which one or more nucleotides (preferably one nucleotide) are replaced with unnatural base nucleotides in the sequence shown in SEQ ID NO.1, and the replacement of the unnatural base nucleotides occurs at positions 12, 14, 17, or 19 at the 5'-end of SEQ ID NO.1;

[0893] The antisense strand 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 -3'(SEQ ID NO.2), where Y 1 and Y 2 is preferably U, and Y 3 is preferably U or T.

[0894] In some embodiments, the sense strand is 5'-CUUGCUACCCAUUAGGAUX 1 -3'(SEQ ID NO.1), where X 1 is selected from A or U, preferably A, and the sense strand comprises a sequence in which one or more nucleotides (preferably one nucleotide) are replaced with unnatural base nucleotides in the sequence shown in SEQ ID NO.1, and the replacement of the unnatural base nucleotides occurs at positions 12, 14, 17, or 19 at the 5'-end of SEQ ID NO.1;

[0895] The antisense strand 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 -3'(SEQ ID NO.2), where Y 1 and Y 2 is preferably U, and the antisense strand may or may not contain Y3 Y 3 It can be U; preferably, in Y 3 When it is not present, the antisense chain can contain additional sequences, such as dT.

[0896] In some implementations, the justice chain 5'-CUUGCUACCCAUUAGGAUX 1 The nucleotides at positions 1, 2, 3, 4, 6, 10, 11, 12, 13, 14, 15, 16, 17, 18, and 19 of the 5' end of the -3'(SEQ ID NO.1) each independently contain a 2'-OMe modification, preferably, X 1 Selected from A or U, preferably A;

[0897] The fifth nucleotide at the 5' end of the positive strand contains a 2'-OMe modification.

[0898] Two of the nucleotides at positions 7, 8, and 9 of the 5' end of the positive strand contain a 2'-F modification, and the remaining one contains either a 2'-OMe modification or a 2'-deoxy modification.

[0899] The positive strand comprises a sequence in which one or more nucleotides (preferably one nucleotide) are replaced by non-natural base nucleotides in the sequence shown in SEQ ID NO.1, wherein the non-natural base nucleotide replacement occurs at position 12, 14, 17 or 19 at the 5' end of SEQ ID NO.1;

[0900] The antisense chain 5'-Y 1 AUCCUAAUGGGUAGCAAGY 2 Y 3 The nucleotides at positions 2 and 14 of the 5' end of the -3' (SEQ ID NO.2) each independently contain a 2'-F modification.

[0901] The nucleotide at the 5' end of the antisense strand contains a 2'-OMe modification.

[0902] The antisense strand is modified with 2'-OMe at positions 15, 17, 18, 19, 20 and 21 of the 5' end, and the nucleotide at position 16 contains a 2'-F modification; wherein, one of the nucleotides contains a 2'-deoxy modification, preferably at position 21 (especially dT).

[0903] In some embodiments, as used herein, in addition to the ribogroup modifications mentioned herein, phosphate backbone modifications are also included. In some embodiments, the sense or antisense strand optionally includes an additional sequence. The phosphate backbone modifications, additional sequences, and ligands are as described herein.

[0904] In some embodiments, as used in this application, in addition to the ribose group modification mentioned in this application, thiophosphate bond (PS) modification is also included, and the number and position of the modification are as described in this application.

[0905] In some embodiments, this application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the nucleotide sequence of the sense or antisense strand of the dsRNA is selected from Tables 13A, 13B, 13C, or 13D below:

[0906] Table 13A.

[0907] Table 13B.

[0908] Table 13C.

[0909] Table 13D.

[0910] Wherein, L represents a ligand, which is as described above; in some embodiments, the ligand is selected from L01 ligand or L02 ligand; wherein, the sequence ID number of the positive strand refers to the sequence ID number of the nucleic acid sequence in the positive strand; the L at the 3' end of the positive strand sequence of the dsRNA ligand conjugate indicates that a ligand is attached at the 3' end of the positive strand sequence in the dsRNA ligand conjugate. Wherein, both uppercase X and lowercase x represent nucleotides having the structure shown by base X, wherein lowercase x indicates that the sugar ring of the nucleotide also has a 2'-OMe modification.

[0911] In another aspect, this application provides a double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, and a pharmaceutically acceptable salt thereof, wherein the double-stranded ribonucleic acid (dsRNA) is as described in this application, and the pharmaceutically acceptable salt is as described in this application.

[0912] In another aspect, this application provides a double-stranded ribonucleic acid (dsRNA) or a ligand conjugate thereof that targets PNPLA3 mRNA, wherein the double-stranded ribonucleic acid (dsRNA) is as described in this application, and the ligand is as described in this application.

[0913] In another aspect, this application provides a pharmaceutical composition targeting PNPLA3 mRNA, comprising the dsRNA of this application, a pharmaceutically acceptable salt thereof or a ligand conjugate thereof; and optionally a pharmaceutically acceptable excipient (carrier or excipient).

[0914] In another aspect, this application provides a kit for treating and / or preventing diseases associated with PNPLA3, comprising the dsRNA of this application, a pharmaceutically acceptable salt thereof, a ligand conjugate thereof, or a pharmaceutical composition thereof; and optionally, instructions for use.

[0915] In another aspect, this application provides a method for treating and / or preventing diseases associated with PNPLA3, comprising administering a therapeutically effective amount of the application's dsRNA, a pharmaceutically acceptable salt thereof, a ligand conjugate thereof, or a pharmaceutical composition to a subject (hereinafter also referred to as a subject).

[0916] On the other hand, this application provides the use of the dsRNA described herein, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition in the preparation of medicaments for the treatment and / or prevention of diseases associated with PNPLA3.

[0917] On the other hand, this application provides the use of the dsRNA described herein, its pharmaceutically acceptable salts, its ligand conjugates, or pharmaceutical compositions in the treatment and / or prevention of diseases associated with PNPLA3.

[0918] On the other hand, this application provides the dsRNA of this application, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition for the treatment and / or prevention of diseases associated with PNPLA3.

[0919] On another front, this application provides a kit for treating and / or preventing diseases, comprising the dsRNA of this application, a pharmaceutically acceptable salt thereof, a ligand conjugate thereof, or a pharmaceutical composition thereof; and optionally, instructions for use.

[0920] In another aspect, this application provides a method for treating and / or preventing a disease, comprising administering a therapeutically effective amount of the application's dsRNA, a pharmaceutically acceptable salt thereof, a ligand conjugate thereof, or a pharmaceutical composition to a subject (hereinafter also referred to as a subject).

[0921] On the other hand, this application provides the use of the dsRNA described herein, its pharmaceutically acceptable salts, its ligand conjugates, or pharmaceutical compositions in the preparation of medicaments for treating and / or preventing diseases.

[0922] On the other hand, this application provides the use of the dsRNA described herein, its pharmaceutically acceptable salts, its ligand conjugates, or pharmaceutical compositions in the treatment and / or prevention of diseases.

[0923] On the other hand, this application provides the dsRNA of this application, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition for the treatment and / or prevention of diseases.

[0924] In some embodiments, the PNPLA3-related diseases include non-alcoholic fatty liver disease.

[0925] In some embodiments, the PNPLA3-related diseases include simple fatty liver (steatohepatitis), non-alcoholic steatohepatitis, cirrhosis (irreversible late-stage liver scarring), or PNPLA3-related obesity.

[0926] In some embodiments, the disease includes non-alcoholic fatty liver disease.

[0927] In some embodiments, the disease includes simple fatty liver (steatohepatitis), non-alcoholic steatohepatitis, cirrhosis (irreversible late-stage scarring of the liver), or obesity.

[0928] In some embodiments, the dsRNA of this application, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition is used as a single therapeutic agent for treating and / or preventing diseases associated with PNPLA3.

[0929] In some embodiments, the dsRNA of this application, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition may be used in combination with other therapeutic agents for the treatment and / or prevention of diseases associated with PNPLA3.

[0930] Pharmaceutically acceptable salts

[0931] In some embodiments, the salts described above are selected from alkali addition salts, acid addition salts, and combinations thereof.

[0932] In some embodiments, the base addition salt is selected from sodium, potassium, calcium, ammonium, organic amine, magnesium salts and combinations thereof, and the acid addition salt is selected from inorganic acid salts, organic acid salts and combinations thereof.

[0933] In some embodiments, the inorganic acid is selected from hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, and combinations thereof, and the organic acid is selected from acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and combinations thereof. Beneficial effects

[0934] This application provides chemical modification patterns for dsRNA that improve the efficacy, stability, duration of action, and / or safety of dsRNA gene silencing activity. The chemical modification patterns for dsRNA described in this application can be universally applied to double-stranded ribonucleic acids with different sequences and targets.

[0935] The double-stranded ribonucleic acid, its pharmaceutically acceptable salt, or its ligand conjugates disclosed in this application have good PNPLA3 inhibitory activity.

[0936] The double-stranded ribonucleic acid (BRNA), its pharmaceutically acceptable salt, or its ligand conjugates disclosed in this application exhibit good PNPLA3 inhibitory activity, in vitro silencing activity, and in vivo (AAV-PNPLA3 mice) inhibition or reduction of PNPLA3 mRNA expression. The BRNA, its pharmaceutically acceptable salt, or its ligand conjugates disclosed in this application demonstrate good stability and can exert a sustained effect in vivo over a long period. Furthermore, the BRNA, its pharmaceutically acceptable salt, or its ligand conjugates disclosed in this application exhibit low off-target effects, minimal toxicity, and high safety. Therefore, they possess promising potential for drug development.

[0937] Definitions and Explanations

[0938] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in accordance with the meaning as understood by one of ordinary skill in the art. When trade names appear herein, they are intended to refer to the corresponding product or its active ingredient.

[0939] The terms "optional" or "optionally" mean that the event or situation subsequently described may or may not occur, and the description includes both the occurrence and non-occurrence of the event or situation. For example, "optionally includes" means to include or not include; "optionally modified" means modified or unmodified.

[0940] In this article, "one or more" refers to an integer from one to fifty (or from one to ten). For example, "one or more" means one, two, three, four, five, six, seven, eight, nine, or ten; or, "one or more" means one, two, three, four, five, or six; or, "one or more" means one, two, or three.

[0941] In this application, unless the context clearly indicates otherwise, “or” means to select one, two or more options, including “and”, but not equivalent to “and”.

[0942] In this application, unless otherwise stated, the terms "comprising, including, and containing" or equivalents are open-ended expressions, meaning that in addition to the listed elements, components, or steps, other unspecified elements, components, or steps may be covered.

[0943] In this application, the phrase "the sense strand or antisense strand is optionally modified" means that any nucleotide of the sense strand or antisense strand is optionally modified.

[0944] When any variable appears more than once in a compound, nucleotide, single-stranded, or double-stranded structure, its definition is independent in each case. For example, the modifications described in this application occur independently; that is, unless otherwise specified, a modification of the sense strand does not affect a modification of the antisense strand, a modification of one nucleotide does not affect a modification of another nucleotide, and a modification of the sugar group of one nucleotide does not affect other modifications of the same nucleotide (e.g., modifications at another location on the sugar group of the same nucleotide, base modifications of the same nucleotide, phosphate backbone modifications of the same nucleotide, etc.). The effects include whether a modification has occurred and the type of modification used.

[0945] As is known in the art, the terms "interfering RNA" or "RNAi" or "interfering RNA sequence" refer to single-stranded RNA (e.g., mature miRNA) or double-stranded RNA (e.g., double-stranded RNA such as siRNA, aiRNA, or pre-miRNA) that, when in the same cell as a target gene or sequence, are capable of reducing or inhibiting the expression of that target gene or sequence (e.g., by mediating the degradation of mRNA complementary to the interfering RNA sequence or inhibiting the translation or transcription of mRNA complementary to the interfering RNA sequence). The interfering RNA may have substantially or completely identical identity to the target gene or sequence, or may include mismatched regions (i.e., mismatched sequences).

[0946] The double-stranded RNA in this application functions as interfering RNA. In this application, the double-stranded RNA is sometimes referred to as siRNA.

[0947] As is known in the art, the terms "mismatch region," "mismatch sequence," or "base mismatch" refer to a portion of an interfering RNA (e.g., siRNA, aiRNA, miRNA) sequence that is not 100% complementary to its target sequence. Interfering RNA (e.g., siRNA, aiRNA, miRNA) may have at least 1, 2, 3, 4, 5, 6, or more mismatch regions. Mismatch regions may be continuous or separated by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more nucleotides. A mismatch region may comprise a single nucleotide or may comprise 2, 3, 4, 5, 6, or more nucleotides.

[0948] The term "identity" refers to the similarity between two nucleotide sequences or two amino acid sequences. Sequence identity preferably involves the percentage of nucleotides or amino acids in a sequence that occupy the same position in two or more sequences of the same length. Specifically, the "% identity" of two amino acid sequences or two nucleotide sequences can be determined by aligning the sequences for optimal comparison (e.g., introducing vacancies in either sequence to achieve optimal alignment with the other) and comparing the amino acids or nucleotides at the corresponding positions. Vacancies are generally considered dissimilar positions, regardless of their actual position in the alignment. An "optimal alignment" is generally the alignment of the two sequences that results in the highest percentage of identity. The percentage of identity is determined by the number of identical nucleotides in the compared sequences (i.e., % identity = number of identical positions / total number of positions × 100). The sequence identity of this application is at least 80%, 85%, 90%, or 95%, preferably at least 90%. Non-limiting examples include: 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and 100%. The percentage of identity between two sequences can be determined using mathematical algorithms known to those skilled in the art.

[0949] As is known in the art, interfering RNA includes "small interfering RNA (siRNA)" with lengths, for example, about 14-40, 15-60, 15-50, 15-40, 15-35, 15-30, 15-25, 18-30, 19-25, 19-23, or 19-21 nucleotides. The double-stranded ribonucleic acid of this application can function as such siRNA and can have the corresponding length.

[0950] As used in this application, for a particular sequence, the nucleotide positions of the sense or antisense strand are counted starting from the 5' end (unless otherwise specified), for example as follows:

[0951] For example, for the sequence shown in equation (I), if x is selected from 0, then the sequence shown in equation (I) is 5'-N1N2N3N4N5N6N7N8N9N10N11N12N13N14N15N16N17N18N19-3', where the first bit of the 5' end of this sequence is N1, the second bit is N2, the third bit is N3, and so on up to the 17th bit is N17, the 18th bit is N18, and the 19th bit is N19;

[0952] For example, for the sequence shown in equation (I), if x is selected from 1, then the sequence shown in equation (I) is 5'-Na1N1N2N3N4N5N6N7N8N9N10N11N12N13N14N15N16N17N18N19-3', where the first position of the 5' end of this sequence is Na1, the second position is N1, the third position is N2, and so on up to the 18th position is N17, the 19th position is N18, and the 20th position is N19;

[0953] For example, for the sequence shown in equation (I), if x is selected from 2, then the sequence shown in equation (I) is 5'-Na2Na1N1N2N3N4N5N6N7N8N9N10N11N12N13N14N15N16N17N18N19-3', where the first position of the 5' end of this sequence is Na2, the second position is Na1, the third position is N1, and so on up to the 19th position is N17, the 20th position is N18, and the 21st position is N19;

[0954] For example, for the sequence shown in equation (II), if y is selected from 0, then the sequence shown in equation (II) is 5'-N1'N2'N3'N4'N5'N6'N7'N8'N9'N10'N11'N12'N13'N14'N15'N16'N17'N18'N19'N20'N21'-3'. In this sequence, the first bit of the 5' end is N1', the second bit is N2', the third bit is N3', and so on up to the 19th bit is N19', the 20th bit is N20', and the 21st bit is N21'.

[0955] For example, for the sequence shown in equation (II), if y is selected from 1, then the sequence shown in equation (II) is 5'-N1'N2'N3'N4'N5'N6'N7'N8'N9'N10'N11'N12'N13'N14'N15'N16'N17'N18'N19'N20'N21'Nb1-3', where the first bit of the 5' end of this sequence is N1', the second bit is N2', the third bit is N3', and so on up to the 20th bit is N20', the 21st bit is N21', and the 22nd bit is Nb1;

[0956] For example, for the sequence shown in equation (II), if y is selected from 2, then the sequence shown in equation (II) is 5'-N1'N2'N3'N4'N5'N6'N7'N8'N9'N10'N11'N12'N13'N14'N15'N16'N17'N18'N19'N20'N21'Nb1Nb2-3', where the first bit of the 5' end of this sequence is N1', the second bit is N2', the third bit is N3', and so on up to the 21st bit is N21', the 22nd bit is Nb1, and the 23rd bit is Nb2;

[0957] For example, for dsRNAs with a 19-nucleotide sense strand and a 21-nucleotide antisense strand, such as 5'-CUUGCUACCCAUUAGGAUA-3', the 5' end has C at position 1, U at position 2, U at position 3, and so on up to A at position 17, U at position 18, and A at position 19; and 5'-UAUCCUAAUGGGUAGCAAGUU-3' has U at position 1, A at position 2, U at position 3, and so on up to G at position 19, U at position 20, and U at position 21.

[0958] If an additional sequence or overhang is joined at the end (5' end or 3' end) of a particular sequence to form a new sequence, or if a particular sequence is included in a longer sequence to form a new sequence, the nucleotide positions of the new sequence are counted starting from the 5' end, and the nucleotide positions of the new sequence are independent of the positions of the particular sequence, that is, the nucleotide positions of the new sequence counted starting from the 5' end do not affect the nucleotide positions of the particular sequence counted starting from the 5' end.

[0959] As is known in the art, and unless otherwise stated, when the term "complementary" is used to describe the relationship between a first nucleic acid sequence and a second nucleic acid sequence, it refers to the ability of an oligonucleotide or polynucleotide containing the first nucleic acid sequence to hybridize with an oligonucleotide or polynucleotide containing the second nucleic acid sequence under specific conditions and form a double-stranded structure. As described in this application, a "complementary" sequence may also include a double-stranded structure formed by base pairs consisting of non-Watson-Crick base pairs and / or non-natural or modified nucleotides, or it may be a double-stranded structure formed entirely by base pairs consisting of non-Watson-Crick base pairs and / or non-natural or modified nucleotides, provided that the above requirements regarding their hybridization ability are met.

[0960] As is known in the art, a “fully complementary” sequence comprises an oligonucleotide or polynucleotide containing a first nucleic acid sequence and an oligonucleotide or polynucleotide containing a second nucleic acid sequence, with base pairing along the full length of both the first and second nucleic acid sequences.

[0961] As is known in the art, “fundamentally complementary” means that two nucleic acid sequences are completely complementary or that at least 85% (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of their overlapping nucleotides are complementary; or that two nucleic acid sequences are completely complementary or that there is a mismatch of 1 to 5 bases.

[0962] The terms “complementary,” “fully complementary,” and “fundamentally complementary” used in this application may be used based on the base pairing between the sense and antisense strands of the dsRNA, or between the antisense strand of the dsRNA and the target sequence, as can be understood from the content in which they are used.

[0963] As is known in the art, the terms "double-stranded ribonucleic acid," "double-stranded RNA," or "dsRNA" are used interchangeably. The term "dsRNA" comprises two antiparallel and complementary nucleic acid strands with a "sense" or "antisense" orientation relative to the target RNA (e.g., the PNPLA3 gene). In embodiments of this application, dsRNA can degrade the target RNA (e.g., mRNA) via RNA interference (RNAi) mechanisms.

[0964] The double-stranded RNA of this application contains a sense strand and an antisense strand. The term "sense strand" or "sensory strand" refers to a single strand in the dsRNA double helix that is substantially complementary to a region of the antisense strand. The terms "antisense strand," "guide strand," or "guide strand" refer to a single strand in the dsRNA double helix that is substantially complementary to a region of the target sequence. If the sense strand is not perfectly complementary to the antisense strand, mismatches may occur within the molecule or in the terminal regions. Typically, the most tolerable mismatches are in the terminal regions.

[0965] As is known in the art, the double strands of dsRNA can have the same or different numbers of nucleotides. The length of the double-stranded region formed by complementarity can be any length that allows for the degradation of the target RNA, and the possible lengths are in the range of about 9 to 40 pairs of nucleotides, such as 15 to 30 pairs, 16 to 28 pairs, 19 to 21 pairs, etc.

[0966] As is known in the art, outside the double-stranded region, dsRNA may include one or more nucleotide overhangs, which refer to at least one unpaired nucleotide / nucleoside analogue. For example, an overhang exists when the 3' end of one strand of the dsRNA extends beyond the 5' end of the other strand (or vice versa). The nucleotides at the "overhang" may include 0-5 nucleotides, where "0" indicates no "overhang" and "5" indicates 5 additional nucleotides (i.e., not paired with the other single strand) on a single strand of the dsRNA double strand. These optional "overhangs" may be located at the 5' and / or 3' ends of any single strand of the two strands of the dsRNA. In some embodiments, the "overhang" comprises 0-5 nucleotides. In some embodiments, the "overhang" comprises 0-2 nucleotides. In some embodiments, the "overhang" at the 3' and / or 5' ends of the sense strand of the dsRNA has 0-2 nucleotides. In some embodiments, the "overhang" at the 3' and / or 5' ends of the antisense strand of the dsRNA has 0-2 nucleotides. The nucleotide forming the "protrusion" can be A, G, C, U, or T, or a modified structure thereof. The nucleotide forming the "protrusion" can be U, T, or dT, or a modified structure thereof. In some embodiments, the "protrusion" includes, but is not limited to, "TT," "dTdT," "UU," or their corresponding modified structures, such as 2'-methoxy-modified UU, i.e., uu. In some embodiments, the "protrusion" at the 3' and / or 5' ends of the antisense strand of the dsRNA is substantially complementary to the target RNA. In some embodiments, the "protrusion" at the 3' and / or 5' ends of the antisense strand of the dsRNA is completely complementary to the target RNA. In some embodiments, the "protrusion" at the 3' end of the antisense strand of the dsRNA is completely complementary to the target RNA. In some embodiments, the "protrusion" at the 3' end of the antisense strand of the dsRNA is selected from unmodified or modified GA. The terms "flat," "flat-ended," "blunt," or "blunt-ended" refer to the absence of unpaired nucleotides at the end of the dsRNA, i.e., no nucleotide protrusion. dsRNAs with blunt ends or blunt ends at both ends are dsRNAs that are double-stranded throughout their entire length, meaning that there are no nucleotide overhangs at either end of the molecule.

[0967] In this application, the dsRNA or any single strand thereof is optionally modified, and both unmodified and modified ribonucleic acid are within the scope of protection of this application. Modifications to the dsRNA or any single strand thereof may be located at the 5' and / or 3' ends, nucleotides, or internucleotide linkages, etc., and all such modifications are within the scope of protection of this application. Synthesis or modification can be performed using methods known in the art.

[0968] In this application, the types of nucleotide modifications include, but are not limited to, modifications occurring at the bases, (ribo)glycosyl groups, or linkages between nucleotides (phosphate backbone modifications). The modifications described in this application occur independently; that is, unless otherwise specified, modification of the sense strand does not affect modification of the antisense strand, modification of one nucleotide does not affect modification of another nucleotide, and the type of modification of one nucleotide does not affect other types of modification (e.g., modification of a glycosyl group does not affect other modifications of the same nucleotide, such as modifications at another location on the glycosyl group of the same nucleotide, base modifications of the same nucleotide, phosphate backbone modifications of the same nucleotide, etc.). The effects include whether modification occurs and the type of modification used.

[0969] In this application, a nucleotide may contain one or more types of modifications, such as only a base modification, a (ribo)syl modification, or a phosphate backbone modification; or simultaneously a base modification, a (ribo)syl modification, or a phosphate backbone modification. Unless otherwise indicated, a description or limitation of a certain type of modification contained in a nucleotide does not preclude the denial that the nucleotide may also contain other types of modifications. For example, "the N5 represents a modified nucleotide, the modification comprising a 2'-OMe or a 2'-F modification" means that N5 contains a (ribo)syl 2'-OMe or a 2'-F modification, and optionally also contains other types of modifications (such as a base modification or a phosphate backbone modification).

[0970] In this application, the modified bases or base modifications refer to the removal of bases from natural bases A, U, C, G, and T (i.e., obtaining base-free nucleotides) or the chemical alteration of non-natural bases (the chemical alteration includes, but is not limited to, altering the structure of atoms, functional groups, or functional groups using methods such as substitution, addition, exclusion, cyclization, and cyclization). The modified bases or base modifications are within the scope of this application. It will be understood in the art that modified bases may result in mismatches, which may affect the pairing properties of the sense and antisense strands of dsRNA or the pairing properties of the antisense strand with the target mRNA, thereby potentially having a beneficial effect on the function and role of dsRNA.

[0971] In this application, the modification of the nucleotide includes, but is not limited to, occurring on the (ribo)glycosyl group of the nucleotide, including one or more substituted or de-substituted glycosyl groups, such as the removal of a hydroxyl group from the carbonyl group, or fluorination, amination, alkylation, hydroxyalkylation, or hydroxyalkenylation. The modification on the glycosyl group can occur at various positions on the sugar ring (e.g., at the 1', 2', 3', or 4' positions of the glycosyl group), or as an isonucleotide, a bridged nucleotide, or an acyclic nucleotide. Exemplarily, the modification on the glycosyl group of the nucleotide includes, but is not limited to, 2'-deoxy (dehydroxylation), 2'-fluorination, 2'-amino, 2'-methyl, 2'-ethyl, 2'-methyl-O-methyl, 2'-ethyl-O-methyl, 2'-O-methyl, 2'-O-ethyl, 2'-O-ethyl-O-methyl, or 2'-O-allyl, with the following possible structures:

[0972] Base represents a base.

[0973] In this application, the phosphate backbone modification includes linker phosphate ester modification between nucleotides and terminal phosphate ester modification, etc., wherein the phosphate backbone modification includes substitution or replacement of atoms or functional groups of phosphate ester groups. For example, linker phosphate ester modification includes thiophosphate (PS), dithiophosphate (PS2), methylphosphonate (MP), methoxypropylphosphonate (MOP), or aminophosphonate. The linker phosphate ester modification can occur between nucleotides at any position in the sequence, such as at the 5' end, 3' end, or in the middle of the sequence. The structure includes, but is not limited to, the following (tautomers, free bases, or salts are all within the scope of protection):

[0974] In this context, R is independently selected from H, hydroxyl groups, or 2' modifications on sugar groups, and Base represents a base.

[0975] In this application, the terminal phosphate modification includes 5' and / or 3' end modifications, referring to modifications occurring at the 5' and / or 3' end of dsRNA or any single strand thereof, such as phosphorylation, conjugation, or reverse bonding. The terminal phosphate modification occurring at the 5' or 3' position of the terminal nucleotide of the sequence is equivalent to occurring at the 5' or 3' end of the sequence. Taking the 5' end as an example, it includes, but is not limited to, 5'-phosphate, 5'-methylphosphonate (5'-MP), 5'-thiophosphate (5'-PS), or 5'-(E)-vinylphosphonate (5'-(E)-VP), with the following structure:

[0976] Where Base represents a base, and Z is selected from H, hydroxyl, or 2' modification on a sugar group.

[0977] In this application, the capital letters G, C, A, U, or T typically represent nucleotides containing guanine, cytosine, adenine, uracil, or thymine as bases. The lowercase letters g, c, a, and u represent nucleotides whose corresponding capital letters are modified with a 2'-methoxy group, i.e., g, c, a, and u represent 2'-O-methyl G, 2'-O-methyl C, 2'-O-methyl A, and 2'-O-methyl U, respectively. A capital letter followed by a lowercase f to its right indicates that the corresponding capital letter's nucleotide is modified with a 2'-fluorine group, i.e., Gf, Cf, Af, and Uf represent 2'-fluoro G, 2'-fluoro C, 2'-fluoro A, and 2'-fluoro U, respectively. A capital letter followed by a lowercase d to its left indicates that the nucleotide represented by the capital letter is 2'-deoxy modified. Specifically, dG, dC, dA, dU, or dT represent 2'-deoxy G, 2'-deoxy C, 2'-deoxy A, 2'-deoxy U, or 2'-deoxy T, respectively.

[0978] In this application, when the sequence involves the sense chain, antisense chain, sequence 1, sequence 2, additional sequence, the sequence shown in formula (I), and / or the sequence shown in formula (II), the uppercase letter X represents a sequence containing a base. The nucleotide. The lowercase letter 'x' indicates that the nucleotide represented by its corresponding uppercase letter is modified with 2'-methoxy, that is, 'x' represents 2'-O-methyl X, and its structure is selected from

[0979] In this application, the 2'-deoxy-modified uracil nucleotide (U) includes both nucleotides formed by removing the 2'-hydroxyl group from the ribosome of the uracil nucleotide (2'-deoxyuracil nucleotide, dU) and thymine nucleotides (2'-deoxythymine nucleotide, dT). These two can be used interchangeably and are both within the scope of protection of this application. In other words, both 2'-deoxythymine nucleotide (dT) and 2'-deoxyuracil nucleotide (dU) can be used as 2'-deoxy-modified uracil nucleotide (U), which is also within the scope of protection of this application.

[0980] The lowercase letter 's' indicates that the two nucleotide residues adjacent to 's' are linked by a thiophosphate group. For example, "csu" indicates that the 'c' and 'u' residues are linked by a thiophosphate group. 'VP-' indicates that the nucleotide to the right of the hyphen is a (E)-vinyl phosphate modified nucleotide. For example, "VP-u" indicates a (E)-vinyl phosphate modified 2'-O-methyl U.

[0981] In this application, the letter N represents a nucleotide with an undefined base, and the number or letter to its right is used to distinguish it from other Ns. A lowercase n indicates that the nucleotide is modified with 2'-OMe. A lowercase f to the right of an uppercase N indicates that the nucleotide is modified with 2'-fluorine. A d to the left of an uppercase N indicates that the nucleotide is modified with 2'-deoxy.

[0982] In this application, the ligand is a group linked to dsRNA, comprising a branched group and a linker, wherein the dsRNA, linker, and branched group are sequentially linked (e.g., as shown in Formula 104). The branched group contains at least one (e.g., one, two, three, four, or five) targeting group that is pharmaceutically acceptable, targeting the dsRNA to a specific tissue or enhancing cellular uptake. The targeting group is, for example, but not limited to, a GalNAc (N-acetylgalactosamine, e.g., as shown in Formula 105) group. Multiple targeting groups are linked in series or parallel via the branched group. The GalNAc group can be monovalent, divalent, trivalent, or tetravalent. The terms monovalent, divalent, trivalent, and tetravalent, as used herein, refer to the molar ratio of dsRNA molecules to GalNAc molecules in a dsRNA-ligand conjugate formed by the dsRNA molecule and a ligand containing GalNAc as a targeting group, respectively, being 1:1, 1:2, 1:3, and 1:4. In some embodiments, when the dsRNA of this application is conjugated with a ligand containing GalNAc, the GalNAc molecule is trivalent or tetravalent. In some embodiments, when the dsRNA of this application is conjugated with a ligand containing GalNAc, the GalNAc molecule is trivalent.

[0983] In this application, the ligand can be attached to the phosphate group, 2'-hydroxyl group, 3'-hydroxyl group, or base of the nucleotide. The ligand can be attached to any nucleotide of the dsRNA, including but not limited to the 5' or 3' terminal nucleotide of the sense or antisense strand, or a non-terminal intermediate nucleotide. When the ligand is attached to the end of the dsRNA strand, it can be attached to the phosphate group of the nucleotide; when the ligand is attached to an intermediate nucleotide of the dsRNA, it can be attached to the sugar ring or base of the nucleotide.

[0984] In this application, the types of ligands or their preparation methods may refer to methods known in the art, including but not limited to the ligands and their preparation methods described in WO2009082607, WO2014025805, WO2015006740, and WO2021249484, the entire disclosure of which is incorporated herein by reference. Exemplary ligands include, but are not limited to, L01 or L02 as described above. The L01 ligand described in this application is the same as the L96 ligand in the prior art.

[0985] In this application, unless otherwise stated, "conjugation" refers to the connection between two or more chemical parts, each having a specific function, in a non-covalent or covalent manner; correspondingly, "conjugated compound" refers to a compound formed by the non-covalent or covalent connection between the respective chemical parts. In this application, conjugated compounds connected to each other in a covalent manner are preferred.

[0986] In this application, the ligand is attached to the 5' or 3' end of the sense or antisense strand. Preferably, the ligand is attached to the 5' or 3' end of the sense strand. More preferably, the ligand is attached to the 3' end of the sense strand. Exemplarily, the dsRNA-ligand conjugate formed by the attachment of the dsRNA to the ligand is shown in Formula 102 or Formula 103 below:

[0987] The compounds of this application (e.g., the double-stranded ribonucleic acid of this application, its pharmaceutically acceptable salts, and its ligand conjugates) may exist in specific geometric or stereoisomeric forms, all of which are within the scope of this application. This application envisions all such compounds, including (R)- and (S)-enantiomers, diastereomers, racemic mixtures, and other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this application. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this application.

[0988] Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

[0989] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being in a non-mirror relationship with each other.

[0990] Unless otherwise specified, use wedge-shaped solid line keys. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by a straight solid line key. and straight dashed key The relative configuration of the center of a solid is indicated by a wavy line. Indicates wedge-shaped solid line key or wedge-shaped dashed key Or use wavy lines Indicates a straight solid line key and / or straight dashed key

[0991] Unless otherwise stated, the terms "rich in one isomer," "isomer enrichment," "rich in one enantiomer," or "enantiomer enrichment" mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0992] Unless otherwise stated, the terms "isomer excess" or "enantiomer excess" refer to the difference between the relative percentages of two isomers or two enantiomers. For example, if one isomer or enantiomer is 90% and the other isomer or enantiomer is 10%, then the isomer or enantiomer excess (ee value) is 80%.

[0993] Optically active (R)- and (S)- isomers, as well as D- and L- isomers, can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. To obtain an enantiomer of a compound of this application, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide a pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a salt of the diastereomeric isomer is formed with a suitable optically active acid or base, followed by diastereomeric resolution using conventional methods known in the art, and then the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomeric isomers is typically accomplished by using chromatography employing a chiral stationary phase and optionally combined with chemical derivatization (e.g., from amines to carbamates). The compounds of this application may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, compounds can be labeled with radioactive isotopes, such as tritium ( 3 H), Iodine-125 ( 125 I) or C-14 14 C). For example, deuterium can be used to replace hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds in this application, regardless of whether they are radioactive, are included within the scope of this application.

[0994] The term “treatment” means administering the compound or preparation described in this application (e.g., the double-stranded ribonucleic acid of this application, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition) to improve or eliminate a disease or one or more symptoms associated with said disease, and includes: (i) suppressing the disease or disease state, i.e., curbing its development; (ii) alleviating the disease or disease state, even if the disease or disease state subsides.

[0995] The term “prevention” means administering the compound or preparation described in this application (e.g., the double-stranded RNA of this application, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition) to prevent a disease or one or more symptoms associated with said disease, and includes: preventing the occurrence of the disease or disease state in a subject, particularly when such subjects are susceptible to the disease state but have not yet been diagnosed with the disease state.

[0996] The terms “subject,” “patient,” or “object” are used interchangeably herein and refer to an animal that has become a subject of treatment, observation, or experimentation. In some embodiments, the subject is a mammal, preferably a primate, and more preferably a human.

[0997] The term "therapeutic effective amount" refers to the amount of the compound of this application (e.g., the double-stranded RNA of this application, its pharmaceutically acceptable salt, or its ligand conjugate) used to treat or prevent a particular disease, condition, or disorder; (ii) reduce, improve, or eliminate one or more symptoms of a particular disease, condition, or disorder; or (iii) prevent or delay the onset of one or more symptoms of a particular disease, condition, or disorder described herein. The amount of the compound of this application constituting a "therapeutic effective amount" varies depending on the compound, the disease state and its severity, the route of administration, and the age of the subject to be treated, but may routinely be determined by a person skilled in the art based on their own knowledge and the content of this application.

[0998] The therapeutic dose of the compound in this application may be determined based on factors such as the specific purpose of treatment, the method of administration, the patient's health and condition, and the prescribing physician's judgment. The proportion or concentration of the compound in the pharmaceutical composition may not be fixed and depends on various factors, including dosage, chemical properties (e.g., hydrophobicity), and route of administration. For example, the compound may be provided as a physiologically buffered saline solution containing about 0.1–10% w / v of the compound for parenteral administration. Some typical dosage ranges are from about 1 μg / kg to about 1 g / kg body weight / day. In some embodiments, the dosage range is from about 0.01 mg / kg to about 100 mg / kg body weight / day. The dosage is likely to depend on variables such as the type and severity of the disease or condition, the general health status of the specific patient, the relative biological potency of the selected compound, the excipient formulation, and the route of administration. The effective dose can be obtained by extrapolation from dose-response curves derived from in vitro or animal model testing systems.

[0999] The term "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and other animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[1000] As pharmaceutically acceptable salts, for example, metal salts, ammonium salts, salts formed with organic bases, salts formed with inorganic acids, salts formed with organic acids, and salts formed with basic or acidic amino acids may be mentioned.

[1001] The term "pharmaceutical composition" refers to a mixture of one or more compounds of this application (e.g., the double-stranded ribonucleic acid of this application, its pharmaceutically acceptable salt, or its ligand conjugate) with a pharmaceutically acceptable carrier, excipient, or excipient. The purpose of a pharmaceutical composition is to facilitate the administration of the compound of this application to an organism. In this document, the terms "pharmaceutical composition" and "formulation" have the same meaning and are used interchangeably. Carriers, excipients, or excipients as used herein include any and all solvents, diluents or other liquid excipients, dispersants or suspending agents, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, etc., suitable for the desired particular dosage form. Various carriers, excipients, or excipients used to formulate pharmaceutically acceptable compositions and methods for their preparation may be employed using known techniques. The use of any conventional carrier media, except those incompatible with the compounds of this application (e.g., producing any adverse biological effects or otherwise interacting harmfully with any other component of the pharmaceutically acceptable composition), is covered within the scope of this application. In some specific embodiments, the carriers, excipients or excipients used herein are those commonly used in the field of dsRNA drug delivery.

[1002] The pharmaceutical compositions of this application can be prepared by combining the compounds of this application with suitable pharmaceutically acceptable carriers, excipients or excipients, for example, formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalers, gels, microspheres and aerosols, etc.

[1003] Typical routes of administration for the compounds of this application or their pharmaceutical compositions include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, vaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[1004] The pharmaceutical composition of this application can be manufactured using methods well known in the art, such as conventional mixing, dissolving, granulation, sugar-coated pill making, grinding, emulsification, freeze drying, etc.

[1005] The kit of this application includes the double-stranded ribonucleic acid of this application, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition; and optionally instructions for using the double-stranded ribonucleic acid of this application, its pharmaceutically acceptable salt, its ligand conjugate, or pharmaceutical composition to treat and / or prevent hepatitis B virus infection.

[1006] The solvents used in this application are commercially available.

[1007] Unless otherwise specified, the solvent ratios used in column chromatography and preparative thin-layer silica gel chromatography in this application are all volume ratios.

[1008] It is known in the art that modified nucleotide groups can be introduced into the dsRNA described in this application using nucleoside monomers with corresponding modifications. Those skilled in the art can learn from the prior art about methods for preparing the corresponding modified nucleoside monomers and methods for introducing the modified nucleotide groups into dsRNA. All modified nucleoside monomers can be obtained commercially or prepared using known methods.

[1009] As is known in the art, the desired ribonucleic acid can be obtained through conventional ribonucleic acid preparation methods (e.g., solid-phase synthesis and liquid-phase synthesis), for example, through phosphoramide solid-phase synthesis technology. The preparation method of the double-stranded ribonucleic acid, its pharmaceutically acceptable salt, or its ligand conjugates of this application comprises the following steps: sequentially linking nucleotide monomers in a 3' to 5' orientation according to the nucleotide type or sequence of the sense or antisense strand of the double-stranded ribonucleic acid to synthesize the sense and antisense strands. The linking of each nucleotide monomer includes four steps: deprotection, coupling, capping, oxidation, or sulfidation. Those skilled in the art can use conventional reaction conditions, reagent types and amounts, or adjust them according to experimental conditions to achieve the deprotection, coupling, capping, oxidation, or sulfidation reactions.

[1010] In some embodiments, for the synthesis of ribonucleic acid containing ligands, the ligand can be linked to the ribonucleic acid through a coupling reaction during or after the synthesis of oligonucleotides. Alternatively, the ligand can be linked to a solid support first, and then the nucleoside monomer can be sequentially linked to the ligand-solid support in a 3' to 5' direction.

[1011] Methods for purification and desalting are well known to those skilled in the art. For example, ribonucleic acid (RNA) can be purified by preparative ion chromatography. Alternatively, RNA can be desalted by reversed-phase chromatography or ultrafiltration centrifugation.

[1012] Annealing methods are well known to those skilled in the art. For example, the sense and antisense chains can be mixed in a 1:1 molar ratio, heated to 70-95°C, and then cooled to room temperature to form a double-chain structure.

[1013] During the synthesis process, the concentration of ribonucleic acid can be detected by, for example, ion exchange chromatography, or the molecular weight can be determined by liquid chromatography-mass spectrometry, or the concentration can be determined by micro spectrophotometer to control the synthesis quality. Such detection methods are well known to those skilled in the art.

[1014] Unless otherwise specified, singular terms encompass plural terms, and plural terms encompass singular terms. Unless otherwise specified, the words "a" or "an" mean "at least one" or "at least one". Unless otherwise specified, the use of "or" means "and / or".

[1015] For purposes of description and disclosure, all patents, patent applications, and other identified publications are expressly incorporated herein by reference. These publications are provided solely because their publication predates the filing date of this application. All statements regarding the dates of these documents or representations of their contents are based on information available to the applicant and do not constitute any acknowledgment of the accuracy of the dates or contents of these documents. Furthermore, in any country, any reference to these publications herein does not constitute an endorsement that such publication is part of the general knowledge in the art.

[1016] In this article, DIC represents N,N'-diisopropylcarbodiimide; DMAP represents 4-dimethylaminopyridine; TCA represents trichloroacetic acid; DCM represents dichloromethane; and ACN represents acetonitrile. Detailed Implementation

[1017] The present application is described in detail below through examples, but this does not imply any adverse limitation thereof. The compounds of this application can be prepared by various synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of this application. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of this application without departing from the spirit and scope of this application.

[1018] The prepared double-stranded ribonucleic acid, its pharmaceutically acceptable salt, or its ligand conjugate can be confirmed as the target product by detection methods such as mass spectrometry.

[1019] Reagents and Materials

[1020] Commercially available protecting monomers used in the synthesis include, but are not limited to:

[1021] reagents used in synthesis

[1022] Preparation Example 1: GalNAc ligands (ligands containing GalNAc groups as targeting groups) were attached to a solid support.

[1023] Weigh 1.0 g of amino-CPG (amino loading: 30-40 μmol / g), add 5 mL of decapping agent, react for 1 min, drain the reagent, and repeat the above operation 4 times. Based on a loading of 35 μmol, add 2 eq of GalNAc ligand / acetonitrile solution, then add a condensation reagent consisting of 2.5 eq DIC and 2.5 eq DMAP. React at 25-30℃ for 24 h. After the reaction, filter off the solvent, wash with anhydrous acetonitrile, and then add GalNAc ligand and condensation reagent according to the above dosage, reacting for another 24 h. After the reaction, wash the solid support with anhydrous acetonitrile, add 4 mL of a 1:1 mixture of Capping A and Capping B, react for 2 min, drain, add another 4 mL of the mixture, react for 2 min, drain, wash with anhydrous acetonitrile, and vacuum dry at 30℃ for 1 h for later use. This example uses L01 ligand (or L02 ligand).

[1024] Preparation Example 2: Preparation of Double-Stranded Ribonucleic Acid

[1025] 2.1.1 Synthesis of GalNAc ligand-ssRNA (sense strand)

[1026] On a K&A nucleic acid synthesizer, the aforementioned GalNAc ligand-solid support was placed in a synthesis column, and ssRNA was synthesized using the standard phosphoramide technique. Specific steps included:

[1027] 1) DMT removal (deprotection): First wash the resin twice with acetonitrile, then remove the DMT on the resin with 3% trichloroacetic acid / DCM, and wash with acetonitrile 4 to 5 times.

[1028] 2) Condensation (coupling): Add monomer and condensation reagent to the synthesis column and condense at room temperature for 10 minutes. Wash with acetonitrile 4-5 times.

[1029] 3) Capping: First use Capping A, then use Capping B to cap the unreacted hydroxyl groups. Wash with acetonitrile 4-5 times.

[1030] 4) Oxidation: Add oxidizing agent, oxidize for 2 minutes, then wash with acetonitrile 4 to 5 times.

[1031] Repeat steps 1) to 4) until all sequences are synthesized. Finally, remove DMT with a decapping agent and wash with acetonitrile 4 to 5 times.

[1032] Formation of thiophosphate bonds: Replace the oxidizing agent with a thioating agent and set the thioating time to 10 minutes to complete the formation of thiophosphate bonds.

[1033] 2.1.2 Synthesis of ssRNA (ligandless sense strand)

[1034] On a K&A nucleic acid synthesizer, a universal CPG solid-phase carrier is placed in a synthesis column, and ssRNA is synthesized according to the standard phosphoramide technique. Specific steps include:

[1035] 1) DMT removal (deprotection): The resin is first washed twice with acetonitrile, and then DMT on the resin is removed with 3% trichloroacetic acid / DCM. The resin is washed 4 to 5 times with acetonitrile.

[1036] 2) Condensation (coupling): Add monomer and condensation reagent to the synthesis column and condense at room temperature for 10 minutes. Wash with acetonitrile 4-5 times.

[1037] 3) Capping: First use Capping A, then use Capping B to cap the unreacted hydroxyl groups. Wash with acetonitrile 4-5 times.

[1038] 4) Oxidation: Add oxidizing agent, oxidize for 2 minutes, then wash with acetonitrile 4 to 5 times.

[1039] Repeat steps 1) to 4) until all sequences are synthesized. Finally, remove DMT with a decapping agent and wash with acetonitrile 4 to 5 times.

[1040] Formation of thiophosphate bonds: Replace the oxidizing agent with a thioating agent and set the thioating time to 10 minutes to complete the formation of thiophosphate bonds.

[1041] 2.2 Synthesis of asRNA (ligandless antisense strand)

[1042] On a K&A nucleic acid synthesizer, the above-mentioned CPG solid-phase carrier was placed into a synthesis column and synthesized according to the method described in 2.1.1 or 2.1.2.

[1043] For the sense and / or antisense chains containing x, the appropriate phosphoramide monomers are synthesized according to the sequence shown using standard phosphoramide techniques.

[1044] 2.3 Separation of ssRNA or asRNA from the solid-phase carrier

[1045] The lysis reagent was ammonia water:ethanol = 3:1. 5 ml of the lysis reagent was added per 100 mg of solid support, and the reaction was stirred at 65-70 °C for 3 h. After the reaction was complete, the mixture was allowed to cool to room temperature and precipitated with ice-cold n-butanol, then placed in a -20 °C freezer for 30 min. The precipitate was obtained by centrifugation, followed by washing with n-butanol, repeated twice. Finally, the precipitate was washed with acetone, centrifuged, and vacuum dried to obtain the crude product, which was then analyzed by mass spectrometry.

[1046] The LC-MS (negative ion mode) conditions are as follows:

[1047] Chromatographic column: HILIC chromatographic column

[1048] Mobile phase A: 20 mm ammonium formate, pH 6.2

[1049] Mobile phase B: 20 ​​mmol ammonium formate + 95% ACN

[1050] Column temperature: 45℃

[1051] Flow rate: 1 mL / min.

[1052] 2.4 Annealing to form siRNA (dsRNA)

[1053] Dissolve the crude single-stranded RNA in water and remove the vector by membrane filtration. Determine the concentrations of the sense and antisense strands using a micro spectrophotometer, add the sense and antisense strands (to a molar ratio of 1:1), mix, denature at 94°C for 4 min, and anneal at room temperature.

[1054] 2.5 Purification of siRNA (dsRNA)

[1055] Purification was performed using a DNAPac RP 10*150mm 4μm column. Mobile phase A was 0.1mol / L triethylamine, pH 8.0, and mobile phase B was 0.1mol / L triethylamine, pH 8.0 + 50% ACN. Detection wavelengths were 215nm and 260nm. The main peak was collected, and rotary evaporation was used to remove most of the solvent to obtain the target product.

[1056] 2.6 Desalination and content calibration

[1057] Using 3K ultrafiltration centrifuge tubes, centrifuge at 12000×g for 12min to concentrate and desalinate the liquid; after the concentrated liquid is ready, water is added to replace it, and this process is repeated more than 5 times.

[1058] Concentration determination: The concentration of the concentrated sample was determined using a micro spectrophotometer.

[1059] Double-stranded ribonucleic acid (dsRNA), its pharmaceutically acceptable salts, or their ligand conjugates are shown in Tables 13A, 13B, 13C, and 13D below:

[1060] Table 13A.

[1061] Table 13B.

[1062] Table 13C.

[1063] Table 13D.

[1064] In this context, the uppercase letters G, C, A, U, and T typically represent nucleotides containing guanine, cytosine, adenine, uracil, and thymine as bases, respectively. The lowercase letters g, c, a, and u represent nucleotides whose ribonucleotides are modified with 2'-OMe, i.e., g, c, a, and u represent 2'-O-methyl G, 2'-O-methyl C, 2'-O-methyl A, and 2'-O-methyl U, respectively. The uppercase letter followed by the lowercase letter f to its right indicates its corresponding... The uppercase letters represent nucleotides whose ribonucleotides are 2'-fluorinated, i.e., Gf, Cf, Af, and Uf represent 2'-fluoroG, 2'-fluoroC, 2'-fluoroA, and 2'-fluoroU, respectively; uppercase letters followed by lowercase 'd' to their left indicate that the corresponding nucleotides are 2'-deoxy, i.e., dG, dC, dA, and dT represent 2'-deoxyG, 2'-deoxyC, 2'-deoxyA, 2'-deoxyU, or 2'-deoxyT, respectively; the uppercase letter X indicates the presence of a base. The nucleotide; the lowercase letter x indicates that the corresponding uppercase letter represents a nucleotide modified by 2'-methoxy, that is, x represents 2'-O-methyl X, and its structure is shown on the right. The lowercase letter 's' indicates that the two nucleotide residues adjacent to 's' are linked by a phosphate thioester group. For example, "csu" indicates that the 'c' and 'u' residues are linked by a phosphate thioester group. 'L' represents a ligand. For example, the structures of ligands L01 and L02 are shown below:

[1065] For example, TDPN2M328L01 indicates that the L ligand in TDPN2M328L is L01, and other cases are similar.

[1066] The aforementioned double-stranded ribonucleic acid, its pharmaceutically acceptable salt, or its ligand conjugate (the ligand is selected from L01 ligand or L02 ligand) can be prepared according to the method described in this application or methods known in the art.

[1067] The mass spectrometry data of the double-stranded ribonucleic acid, its pharmaceutically acceptable salt, or its ligand conjugates obtained in this application are as expected, confirming that the target product has been obtained.

[1068] Experimental Example 2: siRNA-mediated in vitro gene silencing in primary mouse liver cells

[1069] SPF-grade C57BL / 6 mice, aged 4-6 weeks, were purchased from Shanghai Slack Laboratory Animal Co., Ltd., and injected via tail vein with 1×10 11 The AAV-CTM-PNPLA3-I148M virus (human full-length PNPLA3 gene, purchased from Shandong Weizhen Biotechnology Co., Ltd.) was used to anesthetize mice with Virbac 50 (8V8KA) 14 days later, and liver perfusion was performed. The perfusion fluid was purchased from Liver Biotechnolog. After perfusion, the liver capsule was torn open with forceps, and the hepatocytes were gently shaken out to form a milky suspension. The suspension was filtered through a 70μm (200 mesh) sterile sieve and centrifuged twice. The extracted mouse primary hepatocytes were selected and pre-coated with 0.25mL / well collagen (Corning, 354236) in 48-well plates (Corning, 354236). After 5 minutes, the collagen was poured out, and the cell suspension was diluted to 4×10⁻⁶. 5 Cell density of cells / mL, at 1×10 5 Seed 0.25 mL of the prepared siRNA ligand conjugate into 48-well plates. Add 5 μL of the prepared siRNA ligand conjugate to each well of the cell-containing culture plate and incubate at 37°C for 48 h. Extract cellular RNA using an automated nucleic acid extractor (ALLSHENG, Auto-Pure96) and a universal RNA extraction kit using magnetic beads (ONREW, RNC643-03H). Add 200 μL of lysis buffer PLB to the cultured cells, vortex or pipette to disperse the cells, incubate at room temperature for five minutes, then start the program. Follow the prompts to place the sample or reagent into the corresponding slot of the KingFisher Flex nucleic acid extractor.

[1070] After extraction, the extracted mRNA was reverse transcribed using 5×PrimeScript Master Mix (TAKARA, RR036A-1) and the experimental procedure was performed according to the manufacturer's instructions. The reverse transcription reaction was carried out by gently mixing the mixture under the following conditions: 37℃ for 15 min (reverse transcription reaction), 85℃ for 5 sec (reverse transcriptase inactivation reaction), and incubation at 4℃.

[1071] Next, qPCR experiments were performed using the 2×ChamQ SYBR qPCR kit (Vazyme, Q331-02-AA). After the reagents in the kit were completely melted, the mixture was stirred and then centrifuged briefly. qPCR detection was then performed using a Roche Lightcycle-480 real-time PCR instrument.

[1072] The primer sequences used in the PCR process are as follows:

[1073] Human PNPLA3-Foward: 5'-CCTTGGTATGTTCCTGCTTCA-3' (SEQ ID NO. 44);

[1074] Human PNPLA3-Reverse: 5'-ACACGGTGATGGTTGTTTTGG-3' (SEQ ID NO. 45);

[1075] Mouse GAPDH-Forward: 5'-TGCACCACCAACTGCTTAG-3' (SEQ ID NO. 46);

[1076] Mouse GAPDH-Reverse: 5'-GGATGCAGGATGATGTTC-3' (SEQ ID NO. 47).

[1077] The expression level of PNPLA3 mRNA was calculated using a relative quantitative method, namely the ΔΔCt value method.

[1078] The calculation method is as follows: ΔCt(siRNA)=Ct(PNPLA3)–Ct(GAPDH) ΔCt(NC)=Ct(PNPLA3)–Ct(GAPDH) ΔΔCt(siRNA)=ΔCt(siRNA)-ΔCt(average NC) ΔΔCt(NC)=ΔCt(NC)-ΔCt(average NC) Relative expression level of PNPLA3 mRNA=2^(-ΔΔCt(siRNA)) Silencing efficiency of siRNA on PNPLA3 mRNA=(1-2^(-ΔΔCt(siRNA)))×100%.

[1079] The calculated inhibition rates of the siRNA ligand conjugates are shown in Table 14 below.

[1080] Table 14. Determination of silencing efficiency of siRNA ligand conjugates in mouse primary liver cells

[1081] Experimental Example 3: siRNA-mediated silencing of PNPLA3 in mice

[1082] The viral solution (AAV-CTM-hPNPLA3-I148M, Shandong Weizhen Biotechnology Co., Ltd.) was prepared with sterile PBS (pH=7.0) to a concentration of 1×10⁻⁶ before injection. 11 vg / ml solution. 4-6 week old C57BL / 6 mice (purchased from Shanghai Silex Laboratory Animal Co., Ltd.) were each injected via tail vein with 200 μL, which is equivalent to 2 × 10⁶ AAV-CTM-PNPLA3-I148M virus per mouse. 10Vg was administered subcutaneously 7 days later, with a dosage of 10 ml / kg mouse body weight. The day of the first administration was designated as day 0 post-administration. On days 7 and 21 post-administration, 5 mg of liver tissue was collected from each group. After homogenization using a tissue homogenizer (Roche, MagNALyser), total RNA was extracted from the liver tissue using the Takara RNA column extraction kit according to the standard operating procedure for total RNA extraction. After extraction, the extracted RNA was reverse transcribed using 5×PrimeScript Master Mix (TAKARA, RR036A-1) according to the manufacturer's instructions. The reverse transcription reaction was performed after gentle mixing under the following conditions: 37℃ for 15 min (reverse transcription reaction), 85℃ for 5 sec (reverse transcriptase inactivation reaction), and incubation at 4℃.

[1083] Next, qPCR experiments were performed using the 2×ChamQ SYBR qPCR kit (Vazyme, Q331-02-AA). After the reagents in the kit were completely melted, the mixture was stirred and then centrifuged briefly. qPCR detection was then performed using a Roche Lightcycle-480 real-time PCR instrument.

[1084] The primer sequences used in the PCR process are as follows:

[1085] Human PNPLA3-Foward: 5'-CCTTGGTATGTTCCTGCTTCA-3' (SEQ ID NO. 44);

[1086] Human PNPLA3-Reverse: 5'-ACACGGTGATGGTTGTTTTGG-3' (SEQ ID NO. 45);

[1087] Mouse GAPDH-Forward: 5'-TGCACCACCAACTGCTTAG-3' (SEQ ID NO. 46);

[1088] Mouse GAPDH-Reverse: 5'-GGATGCAGGATGATGTTC-3' (SEQ ID NO. 47).

[1089] The expression level of PNPLA3 mRNA was calculated using a relative quantitative method, namely the ΔΔCt value method.

[1090] The calculation method is as follows: ΔCt(siRNA)=Ct(PNPLA3)–Ct(GAPDH) ΔCt(NC)=Ct(PNPLA3)–Ct(GAPDH) ΔΔCt(siRNA)=ΔCt(siRNA)-ΔCt(average NC) ΔΔCt(NC)=ΔCt(NC)-ΔCt(average NC) Relative expression level of PNPLA3 mRNA=2^(-ΔΔCt(siRNA)) Silencing efficiency of siRNA on PNPLA3 mRNA=(1-2^(-ΔΔCt(siRNA)))×100%.

[1091] The calculated inhibition rates of siRNA ligand conjugates in mice are shown in Table 15 below.

[1092] Table 15. Determination of silencing efficiency of siRNA ligand conjugates in mouse liver

[1093] The above experimental results indicate that the double-stranded ribonucleic acid ligand conjugate of this application inhibits or reduces PNPLA3 mRNA expression in vivo (AAV-PNPLA3 mice).

[1094] Experimental Example 4: siRNA-mediated in vitro gene silencing in Hep3B cells

[1095] Hep3B cells were digested with trypsin, and the cell suspension was diluted to 1.5 × 10⁻⁶. 5 A cell density of 1.5 × 10⁶ cells / mL was established. 4 90 μL of siRNA was seeded per well in a 96-well plate (collagen-coated for 5 minutes). 0.3 μL of RNAiMAX per well was added to 10 μL of OPTI-MEM per well and mixed. 0.5 μL of siRNA of different concentrations per well was added to 10 μL of OPTI-MEM containing RNAiMAX per well and mixed. The mixture was incubated at room temperature for 5 minutes. Then, 10 μL of this mixture was added to each well of the 96-well plate and incubated at 37°C with 5% CO2 for 48 hours. After incubation, RNA was extracted from the cells using an automated nucleic acid extractor (ALLSHENG, Auto-Pure96) and a universal RNA extraction kit using magnetic beads (ONREW, RNC643-03H). 200 μL of lysis buffer PLB was added to the cultured cells. The cells were vortexed or dispersed using a pipette. After incubation at room temperature for 5 minutes, the program was started. Following the prompts, the 96-well plate containing the sample or reagent was placed into the corresponding slot of the KingFisher Flex nucleic acid extractor.

[1096] After extraction, the extracted mRNA was reverse transcribed using 5×PrimeScript Master Mix (TAKARA, RR036A-1) and the experimental procedure was performed according to the manufacturer's instructions. The reverse transcription reaction was carried out by gently mixing the mixture under the following conditions: 37℃ for 15 min (reverse transcription reaction), 85℃ for 5 sec (reverse transcriptase inactivation reaction), and incubation at 4℃.

[1097] Next, qPCR experiments were performed using the 2×ChamQ SYBR qPCR kit (Vazyme, Q331-02-AA). After the reagents in the kit were completely melted, the mixture was stirred and then centrifuged briefly. qPCR detection was then performed using a Roche Lightcycle-480 real-time PCR instrument.

[1098] The primer sequences used in the PCR process are shown in Table 16 below.

[1099] Table 16

[1100] The expression level of PNPLA3 mRNA was calculated using a relative quantitative method, namely the ΔΔCt value method.

[1101] The calculation method is as follows: ΔCt(siRNA)=Ct(PNPLA3)–Ct(GAPDH) ΔCt(NC)=Ct(PNPLA3)–Ct(GAPDH) ΔΔCt(siRNA)=ΔCt(siRNA)-ΔCt(average NC) ΔΔCt(NC)=ΔCt(NC)-ΔCt(average NC) Relative expression level of PNPLA3 mRNA=2^(-ΔΔCt(siRNA)) Silencing efficiency of siRNA on PNPLA3 mRNA=(1-2^(-ΔΔCt(siRNA)))×100%.

[1102] The test results are shown in Tables 17 and 18.

[1103] Table 17

[1104] Table 18

Claims

1. A double-stranded ribonucleic acid (dsRNA) targeting PNPLA3 mRNA, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, wherein the sense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.1, and the antisense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.2, and the antisense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.2, and the antisense strand contains at least 3 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.

2. 5’-CUUGCUACCCAUUAGGAUX 1 -3’(SEQ ID NO.1), 5′-Y 1 AUCCUAAUGGGUAGCAAGY 2 THE 3 -3'(SEQ ID NO.2): The X 1 Y 1 Y 2 Or Y 3 Each is independently selected from A, U, C, or G. The justice chain or antisense chain is optionally modified.

2. The double-stranded ribonucleic acid (dsRNA) as described in claim 1, its pharmaceutically acceptable salt, or a ligand conjugate thereof, wherein, The antisense strand is at least substantially complementary to the target gene. The sense strand contains sequence 1, and the antisense strand contains sequence 2. Sequence 1 and sequence 2 are at least substantially complementary. Sequence 1 contains the sequence shown in SEQ ID NO. 1, and sequence 2 contains the sequence shown in SEQ ID NO.

1. The sequence 1 has 19 to 21 nucleotides, the sequence 1 contains two or three nucleotides containing 2'-F modification, and the sequence 1 does not contain three consecutive nucleotides containing 2'-F modification; The second sequence has 19 to 23 nucleotides, and contains two, three or four nucleotides containing 2'-F modification, and optionally contains nucleotides containing 2'-deoxy modification.

3. The double-stranded ribonucleic acid (dsRNA) as described in claim 1 or 2, its pharmaceutically acceptable salt, or a ligand conjugate thereof, wherein, Sequence 1 contains two or three nucleotides with 2'-F modification, but sequence 1 does not contain three consecutive nucleotides with 2'-F modification. The nucleotides of sequence 1, other than those containing the 2'-F modified nucleotide, contain 2'-OMe modification or 2'-deoxy modification; or, The nucleotides modified by 2'-F in sequence 1 are located at positions 5 to 11 from the 5' end of sequence 1.

4. The double-stranded ribonucleic acid (dsRNA), its pharmaceutically acceptable salt, or a ligand conjugate thereof as described in any one of claims 1-3, wherein, Sequence 2 contains two, three, or four nucleotides containing a 2'-F modification, and the nucleotides of Sequence 2 other than those containing the 2'-F modification contain a 2'-OMe modification or a 2'-deoxy modification; or, The nucleotides at positions 2 and 14 of the 5' end of sequence 2 contain a 2'-F modification; or, The 6th nucleotide from the 5' end of sequence 2 contains a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification; or, The nucleotide at position 16 of the 5' end of sequence 2 contains a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification; or, One to three nucleotides at positions 15-19, 15-20, 15-21, 15-22, or 15-23 at the 5' end of sequence 2 contain a 2'-deoxy modification; or, The nucleotides at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, and 13 at the 5' end of sequence 2 each independently contain a 2'-OMe modification.

5. The double-stranded ribonucleic acid (dsRNA), its pharmaceutically acceptable salt, or its ligand conjugate as described in any one of claims 1 to 4, wherein, Sequence 1 contains one or more non-natural nucleotide bases; or... One or more nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 at the 5' end of sequence 1 each independently contain a non-natural base nucleotide.

6. The double-stranded ribonucleic acid (dsRNA) as described in any one of claims 1 to 5, its pharmaceutically acceptable salt, or a ligand conjugate thereof, wherein, Sequence 2 contains one or more non-natural nucleotide bases; or... One or more nucleotides at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23 at the 5' end of sequence 2 each independently contain a non-natural base nucleotide.

7. The double-stranded ribonucleic acid (dsRNA) as described in claim 1, its pharmaceutically acceptable salt, or a ligand conjugate thereof, wherein, The dsRNA comprises a sense strand and an antisense strand, each having 19 to 40 nucleotides. The antisense strand is at least substantially complementary to the target gene. The sense strand comprises the sequence shown in formula (I), and the antisense strand comprises the sequence shown in formula (II). Formula (I): 5'-(Na) x N1N2N3N4N5N6N7N8N9N10N11N12N13N14N15N16N17N18N19-3', Equation (II): 5'-N1'N2'N3'N4'N5'N6'N7'N8'N9'N10'N11'N12'N13'N14'N15'N16'N17'N18'N19'N20'N21'(Nb) y -3'; Each of Na, N1, N2, N3, N4, N6, N10, N11, N12, N13, N14, N15, N16, N17, N18, and N19 independently represents an unmodified or modified nucleotide. The N5 represents a modified nucleotide, the modification comprising either a 2'-OMe modification or a 2'-F modification. N7, N8, and N9 each independently represent a modified nucleotide, the modification including 2'-OMe modification, 2-F modification, or 2'-deoxy modification, and not all of N7, N8, and N9 are selected from 2'-F modification. The x is selected from 0, 1, or 2; The N1', N3', N4', N5', N7', N8', N9', N10', N11', N12', and N13' each independently represent an unmodified or modified nucleotide. N2' and N14' each independently represent a modified nucleotide. The N6' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification. The N15', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification. The N16' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification. The value of y is selected from 0, 1, or 2.

8. The double-stranded ribonucleic acid (dsRNA) of claim 7, its pharmaceutically acceptable salt, or a ligand conjugate thereof, wherein, Each of Na, N1, N2, N3, N4, N6, N10, N11, N12, N13, N14, N15, N16, N17, N18, and N19 independently represents a modified nucleotide, which does not include a 2'-F modification; or, The N5 represents a modified nucleotide, the modification comprising a 2'-OMe modification; or, Each of N7, N8, and N9 independently represents a modified nucleotide, the modification including 2'-OMe modification, 2-F modification, or 2'-deoxy modification, and not all of N7, N8, and N9 are selected from 2'-OMe modification or 2'-F modification; or, One or two of N7, N8, and N9 contain 2'-F modification, and each of the remaining ones independently contains 2'-OMe modification or 2'-deoxygenation modification; or, The x is selected from 0; or, If x is selected from 1, then Na represents a modified nucleotide Na1; or, The x is selected from 2, then Na represents two modified nucleotides Na1Na2, where Na1 is linked to N1.

9. The double-stranded ribonucleic acid (dsRNA) as described in claim 7 or 8, its pharmaceutically acceptable salt, or a ligand conjugate thereof, wherein, The N1', N3', N4', N5', N7', N8', N9', N10', N11', N12', and N13' each independently represent a modified nucleotide. The modification does not include the 2'-F modification; or, The N2' and N14' each independently represent a modified nucleotide, which does not include the 2'-OMe modification; or, The N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide that does not contain a 2'-deoxy modification; or, The N15', N16', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, at least one of which contains a 2'-deoxy modification; or, N15', N17', N18', N19', N20', N21' and each Nb independently represent a modified nucleotide, the modification including 2'-OMe modification or 2'-deoxy modification; N16' represents a modified nucleotide, the modification including 2'-OMe modification, 2'-F modification, or 2'-deoxy modification; wherein one or two nucleotides contain 2'-deoxy modification; or, The y is selected from 0; or, If y is selected from 1, then Nb represents a modified nucleotide Nb1; or, If y is selected from 2, then Nb represents two modified nucleotides Nb1Nb2, where Nb1 is connected to N21'.

10. The double-stranded ribonucleic acid (dsRNA) as described in any one of claims 7 to 9, its pharmaceutically acceptable salt, or a ligand conjugate thereof, wherein, The sequence represented by formula (I) contains one or more non-natural base nucleotides; or, One or more nucleotides in Na1, Na2, N1, N2, N3, N4, N5, N6, N7, N8, N9, N10, N11, N12, N13, N14, N15, N16, N17, N18 or N19 of the sequence shown in Formula (I) each independently contain one or more non-natural base nucleotides.

11. The double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof as described in any one of claims 6 to 9, wherein the sequence represented by formula (II) comprises one or more non-natural base nucleotides; or, One or more of N1', N2', N3', N4', N5', N6', N7', N8', N9', N10', N11', N12', N13', N14', N15', N16', N17', N18', N19', N20', N21', Nb1 or Nb2 of the sequence shown in formula (II) each independently contains one or more non-natural base nucleotides.

12. The double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, as described in any one of claims 1-11, wherein, The sense strand comprises a sequence in which one or more non-natural nucleotides are replaced by non-natural nucleotides in the sequence shown in SEQ ID NO.1, and the antisense strand comprises a sequence in which one or more non-natural nucleotides are replaced by non-natural nucleotides in the sequence shown in SEQ ID NO.

2.

13. The double-stranded ribonucleic acid (dsRNA) as described in claim 1, its pharmaceutically acceptable salt, or a ligand conjugate thereof, wherein, The sense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.1, with a difference of no more than 3 nucleotides; the antisense strand contains at least 15 consecutive nucleotides of the nucleotide sequence shown in SEQ ID NO.2, with a difference of no more than 3 nucleotides. 5’-CUUGCUACCCAUUAGGAUX 1 -3’(SEQ ID NO.1), 5′-Y 1 AUCCUAAUGGGUAGCAAGY 2 THE 3 -3'(SEQ ID NO.2): The X 1 Y 1 Y 2 Or Y 3 Each is independently selected from A, U, C, or G. The nucleotides at positions 1, 2, 3, 4, 6, 10, 11, 12, 13, 14, 15, 16, 17, 18, and 19 of the 5' end of the positive strand are either unmodified or modified nucleotides. The 5th nucleotide at the 5' end of the positive strand contains a 2'-OMe modification or a 2'-F modification. The nucleotides at positions 7, 8, and 9 of the 5' end of the positive strand contain 2'-OMe, 2'-F, or 2'-deoxy modifications, and are not all selected from 2'-F modifications; The nucleotides at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, and 13 of the 5' end of the antisense strand are either unmodified or modified nucleotides. The nucleotides at positions 2 and 14 of the 5' end of the antisense strand are modified nucleotides. The nucleotide at position 6 of the 5' end of the antisense strand contains a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification. The nucleotides at positions 15, 17, 18, 19, 20, and 21 of the 5' end of the antisense strand contain a 2'-OMe modification or a 2'-deoxy modification. The nucleotide at position 16 of the 5' end of the antisense strand contains a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification.

14. The double-stranded ribonucleic acid (dsRNA), its pharmaceutically acceptable salt, or a ligand conjugate thereof as described in any one of claims 1 to 13, wherein, The nucleotides at positions 1, 2, 3, 4, 6, 10, 11, 12, 13, 14, 15, 16, 17, 18, and 19 of the 5' end of the positive strand are modified nucleotides, excluding 2'-F modification; or, The nucleotides at positions 7, 8, and 9 of the 5' end of the positive strand contain a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification, and are not all selected from 2'-OMe modifications or 2'-F modifications; or, One or two of the nucleotides at positions 7, 8, and 9 of the 5' end of the positive strand contain a 2'-F modification, and each of the remaining nucleotides independently contains a 2'-OMe modification or a 2'-deoxy modification.

15. The double-stranded ribonucleic acid (dsRNA), its pharmaceutically acceptable salt, or a ligand conjugate thereof as described in any one of claims 1 to 14, wherein, The nucleotides at positions 1, 3, 4, 5, 7, 8, 9, 10, 11, 12, and 13 of the 5' end of the antisense strand are modified nucleotides, excluding 2'-F modification; or, The nucleotides at positions 2 and 14 of the 5' end of the antisense strand are modified nucleotides, and the modification does not include 2'-OMe modification; or, The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the antisense strand do not contain a 2'-deoxy modification; or, At least one of the nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the antisense strand contains a 2'-deoxy modification; or, The nucleotides at positions 15, 16, 17, 18, 19, 20, and 21 of the 5' end of the antisense strand contain a 2'-OMe modification or a 2'-deoxy modification, and the nucleotide at position 16 contains a 2'-OMe modification, a 2'-F modification, or a 2'-deoxy modification. Two or one of the aforementioned nucleotides contain a 2'-deoxy modification.

16. The double-stranded ribonucleic acid (dsRNA) as described in any one of claims 1 to 15, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, selected from the double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof described in Tables 13A, 13B, 13C, or 13D.

17. The double-stranded ribonucleic acid (dsRNA), a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, as described in any one of claims 5 to 16, wherein, The bases of the non-natural nucleotide are selected from the following structure X:

18. The double-stranded ribonucleic acid (dsRNA) as described in any one of claims 1 to 17, its pharmaceutically acceptable salt, or a ligand conjugate thereof, wherein, The dsRNA includes phosphate backbone modifications; optionally, The phosphate backbone modification includes phosphate ester modification of linking bonds between nucleotides and terminal phosphate ester modification; optionally, The phosphate backbone comprises at least one linking phosphate ester modification, wherein the linking phosphate ester modification includes a thiophosphate bond, a dithiophosphate bond, a methylphosphonate bond, a methoxypropylphosphonate bond, or an aminophosphonate bond; optionally, The sense or antisense chain contains 1 to 10 linking phosphate ester modifications; optionally, The terminal phosphate modification comprises nucleotide 5'-phosphate, 5'-methylphosphonate, 5'-thiophosphate, or 5'-(E)-vinylphosphonate.

19. The double-stranded ribonucleic acid (dsRNA) as described in any one of claims 1 to 18, its pharmaceutically acceptable salt, or a ligand conjugate thereof, wherein, The ligand of the conjugate contains at least one targeting group; or the ligand contains one, two, three, four, or five targeting groups; or, The targeting group is a GalNAc group; or... The ligand includes the following branched groups: or, The ligands are selected from:

20. A pharmaceutical composition comprising, as described in any one of claims 1-19, a pharmaceutically acceptable salt thereof or a ligand conjugate thereof, and optionally a pharmaceutically acceptable excipient.

21. Use of the double-stranded ribonucleic acid (dsRNA) of any one of claims 1-19, a pharmaceutically acceptable salt thereof, or a ligand conjugate thereof, or the pharmaceutical composition of claim 20, in the preparation of a medicament for treating and / or preventing diseases related to a target gene, wherein, The target gene is PNPLA3; or The diseases mentioned include simple fatty liver (steatohepatitis), non-alcoholic steatohepatitis, cirrhosis (irreversible late-stage scarring of the liver), or PNPLA3-related obesity.

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