Modified nucleoside, sirna comprising same, composition, and use method therefor

Abstract

Provided are a modified nucleoside, a use and method for using the nucleoside to prepare an RNAi preparation for extrahepatic delivery, and an RNAi preparation for extrahepatic delivery obtained thereby. The RNAi preparation can effectively deliver RNAi to extrahepatic tissue, such as muscle tissue, like skeletal muscle tissue and myocardial tissue, and adipose tissue.

Description

Modified nucleosides, siRNAs containing them, compositions thereof, and methods of use thereof

[0001] Cross-reference to related applications

[0002] This application is based on and claims priority to Chinese patent applications No. 202411595544.7, filed on November 8, 2024, and No. 202510899205.6, filed on June 30, 2025, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This invention provides a modified nucleoside, the use and method of preparing an RNAi activator using said nucleoside, and the RNAi activator obtained therefrom, which can effectively deliver RNAi to specific tissues or cells, such as muscle tissue (e.g., skeletal muscle tissue or cardiac muscle tissue) or cells, or adipose tissue or cells. Specifically, this invention provides a nucleoside modified with a long-chain hydrocarbon chain and a double-stranded ribonucleic acid (dsRNA) activator prepared using said nucleoside for inhibiting the expression of target genes, compositions comprising such dsRNA activators, and methods of use thereof in subjects treating diseases or conditions related to the genes targeted by said dsRNA activators. Background Technology

[0004] RNA interference (RNAi) is a gene silencing phenomenon induced by RNA (endogenous or exogenous). The mechanism involves short antisense RNAs inhibiting gene expression by blocking the transcription or translation of genes with complementary sequences. RNAi has been confirmed to be widespread in various organisms, but the origin and biosynthesis of core RNAi components differ significantly among species. Currently, it is generally believed that 19–30 nt non-coding small RNAs (sRNAs) play a crucial role in RNAi. sRNAs can be classified into three categories based on their origin, structure, and biological function: small interfering RNAs (siRNAs), microRNAs (miRNAs), and piwiRNAs (piwi-interacting RNAs). In 2001, researchers discovered that the Dicer enzyme is a key enzyme in the RNAi initiation step. Dicer cleaves long dsRNAs into mature siRNAs; then, the siRNA single strand is loaded onto the Argonaute protein, which has endonuclease activity, forming the RNA-induced silencing complex (RISC). After siRNA binds to complementary messenger RNA (mRNA), the target mRNA is either directly cleaved by Argonaute or mediated by RISC to inhibit mRNA translation. Researchers subsequently discovered that RNA-dependent RNA polymerase (RdRP) is involved in the production and replication of secondary small RNAs, further enhancing and amplifying the silencing effect.

[0005] In 2003, siRNA was first used as a drug in mammals. However, the initial clinical trials of siRNAs showed little RNAi efficacy and were accompanied by uncertain interference effects. Immune-related toxicity also occurred during the trials, ultimately leading to failure. Subsequent second-wave clinical trials found that systemic application of siRNA nanoformulations could produce effects in humans, but the efficacy was limited, exhibiting significant dose-limiting toxicity and insufficient therapeutic efficacy. Researchers have made some progress by improving siRNA delivery, sequence selection, chemical formulation, and delivery mechanisms. In 2018, the RNAi drug ONPATTRO (patisiran) received approval from the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMA). This new type of siRNA drug is primarily used to treat hereditary transthyretin amyloidosis.

[0006] Effective delivery of RNAi activators to cells in vivo requires specific targeting and avoidance of the extracellular environment, particularly serum proteins. RNAi-based therapies have shown promising clinical results in the treatment of liver-related conditions. When administered systemically, RNAi activators naturally accumulate in the liver. Similarly, when administered topically, although RNAi activators can achieve significant reductions in target genes, their distribution in muscle or adipose tissue is limited, and reductions in target genes are observed only in a small portion of the tissue, severely limiting their therapeutic applications.

[0007] Therefore, effectively delivering RNAi to specific tissues such as muscle and adipose tissue remains a challenge, and new and improved compositions and methods for in vivo delivery of RNAi active agents are still needed to realize and enhance the therapeutic potential of RNAi active agents. Summary of the Invention

[0008] Invention Summary

[0009] The inventors have discovered that by introducing one or more long hydrocarbon chains by introducing the compounds of the present invention at one or more positions on at least one strand of double-stranded RNA, it is possible to efficiently deliver RNAi to specific tissues or cells (e.g., muscle tissue (such as skeletal muscle tissue or cardiac tissue) or cells, or adipose tissue or cells).

[0010] Therefore, in one aspect, the present invention provides a compound of formula (I) or its subforms such as formulas (Ia) to (If), or a salt thereof, which, when introduced into double-stranded RNA, for example, by adding the backbone structure of the compound to one or more positions of the double-stranded RNA (e.g., at the 5' end and / or the 3' end) and / or by replacing nucleotides at one or more positions of the double-stranded RNA (e.g., internal positions) with the backbone structure of the compound, enables the efficient delivery of RNAi to specific tissues or cells (e.g., muscle tissue (such as skeletal muscle tissue or cardiac muscle tissue) or cells, or adipose tissue or cells).

[0011] The variables are defined as described in the text.

[0012] On the other hand, the present invention provides an RNAi activator that can effectively deliver RNAi to specific tissues or cells, such as muscle tissue (e.g., skeletal muscle tissue or cardiac muscle tissue) or cells, or adipose tissue or cells.

[0013] In some embodiments, the present invention provides a double-stranded RNA (dsRNA) activator for inhibiting the expression of a target gene (e.g., the SOD1 gene or the ACVR1C gene), comprising an antisense strand complementary to the target gene (e.g., the SOD1 gene); a sense strand complementary to the antisense strand and forming a double-stranded region with the antisense strand; and one or more long-chain hydrocarbon chains conjugated to one or more sites (e.g., internal sites) on at least one strand, wherein the dsRNA activator is adapted for delivery to a specific tissue or cell, such as muscle tissue (e.g., skeletal muscle tissue or cardiac muscle tissue) or cells, and adipose tissue or cells.

[0014] In some embodiments, the conjugated one or more long-chain hydrocarbon chains are introduced by adding the backbone structure of one or more compounds of the present invention to the 5' or 3' end of at least one of the sense and / or antisense chains, and / or by replacing nucleotides at one or more internal positions on at least one chain with the backbone structure of one or more compounds of the present invention, thereby introducing the conjugated one or more long-chain hydrocarbon chains onto the nucleotide chain.

[0015] The present invention also provides a pharmaceutical composition comprising the dsRNA activator of the present invention and optionally a pharmaceutically acceptable carrier.

[0016] The present invention also provides a pharmaceutical combination comprising the dsRNA activator of the present invention, and one or more other therapeutic agents effective for the prevention or treatment of diseases or conditions related to the target gene targeted by the dsRNA activator, such as other therapeutic agents for the prevention or treatment of skeletal muscle diseases, cardiomyopathy or adipose tissue diseases, such as other therapeutic agents effective for SOD1 or ACVR1C-related diseases or conditions.

[0017] The present invention also provides the use of the dsRNA activator and / or pharmaceutical composition and / or pharmaceutical combination of the present invention in the preparation of a medicament for the prevention or treatment of diseases or conditions associated with the target gene targeted by the dsRNA activator, such as for the prevention or treatment of skeletal muscle diseases, cardiomyopathy or adipose tissue diseases, such as SOD1 or ACVR1C-related diseases or conditions.

[0018] The present invention also provides a method for preventing or treating diseases or conditions related to the target genes targeted by the dsRNA activator, such as SOD1 or ACVR1C-related diseases or conditions, the method comprising administering an effective amount of the dsRNA activator and / or pharmaceutical composition and / or pharmaceutical combination of the present invention to a subject in need.

[0019] In another aspect, the present invention provides the use of compounds of formula (I) or subforms such as formulas (Ia) to (If) or salts thereof for the preparation of nucleic acid conjugates, such as RNAi activators, such as dsRNA activators, that can be efficiently delivered to specific tissues or cells (e.g., muscle tissue (e.g., skeletal muscle tissue or cardiac tissue) or cells, or adipose tissue or cells).

[0020] In another aspect, the present invention provides a method for preparing nucleic acid conjugates, such as RNAi activators and dsRNA activators, for delivery to specific tissues or cells (e.g., muscle tissue (such as skeletal muscle tissue or cardiac tissue) or cells, or adipose tissue or cells), the method comprising adding the backbone structure of the compound of formula (I) of the present invention at one or more positions of double-stranded RNA (e.g., at the 5' end and / or the 3' end) and / or replacing nucleotides at one or more positions of double-stranded RNA (e.g., internal positions) with the backbone structure of the compound of formula (I) of the present invention.

[0021] Invention Details

[0022] Before describing the invention in detail below, it should be understood that the invention is not limited to the specific methodologies, approaches, and reagents described herein, as these can vary. The materials, methods, and examples described herein are illustrative only and are not intended to limit the scope of the invention, which is defined solely by the appended claims. Furthermore, other features, objects, and advantages of the invention will become apparent from this specification and the accompanying drawings.

[0023] All publications, patent applications, patents and other references mentioned in this article are incorporated in full by way of citation.

[0024] I. Compounds

[0025] In one aspect, the present invention provides a compound that can be incorporated into double-stranded RNA, for example, by adding the backbone structure of the compound at one or more positions (e.g., at the 5' end and / or the 3' end) of the double-stranded RNA and / or replacing nucleotides at one or more positions (e.g., internal positions) of the double-stranded RNA with the backbone structure of the compound, for the purpose of efficiently delivering the double-stranded RNA to a specific tissue or cell (e.g., muscle tissue (such as skeletal muscle tissue or cardiac tissue) or cell, or adipose tissue or cell).

[0026] In some embodiments, the present invention provides compounds of formula (I) or salts thereof (e.g., pharmaceutically acceptable salts), solvates (e.g., hydrates), or isotopically labeled compounds (e.g., deuterated compounds):

[0027] in:

[0028] B is H, a base (including modified or unmodified bases), or -L. B -(C 1-30 (hydrocarbon group), of which L B It is a linker;

[0029] R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support, optionally wherein R1 and R2 are not both C 1-30 hydrocarbon group;

[0030] X is selected from non-existent, -O-, -S-, -SS-, -OC(=O)-, -C(=O)-O-, -NR4C(=O)-, -C(=O)NR4-, -NR4-, -O-NR4-, -NR4-O-, -O-NR4C(=O)- and -C(=O)NR4-O-;

[0031] R3 is H or C. 1-30 hydrocarbon group;

[0032] R B For H or C 1-6 alkyl;

[0033] Or R B Together with XR3, they form C 2-5 An alkylene chain, wherein the alkylene chain is optionally substituted by one or more groups independently selected from the following: hydroxyl, halogen, cyano, mercapto, azide, nitro, NR. a R b C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 1-6 Alkoxy;

[0034] R1' represents H and -C. 0-6 alkylene-O-(C 1-30 hydrocarbon group), -C 0-6 alkylene-S-(C 1-30 hydrocarbon group), -C 0-6 Alkylene-NR4-(C 1-30 hydrocarbon group), -C 0-6 Alkylene-C(O)-(C 1-30 (hydrocarbon group) or -C 0-6 Alkylene-C(S)-(C 1-30 (hydrocarbon group);

[0035] Or R1' is -C 0-6 Alkylene -O-, -C 0-6 Alkylene-S-, -C0-6 Alkylene-NR4-, -C 0-6 alkylene-C(O)- or -C 0-6 Alkylene-C(S)-,XR3 indicates Furthermore, R1' connects with XR3 to form a bridge ring, where the wavy line indicates that the valence bond is connected to R1', and the asterisk indicates that the valence bond is connected to the rest of the molecule;

[0036] R m and R n Each is independently H or C 1-6 Alkyl, or R m and R n The connection forms a single bond;

[0037] R4 is H or C. 1-6 alkyl;

[0038] R a and R b Each is independently H or C 1-6 alkyl;

[0039] Wherein, the C 1-30 The hydrocarbon group is optionally selected from hydroxyl, halogen, cyano, mercapto, azide, nitro, NR a R b C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 3-8 Cyclic hydrocarbon group, C 3-8 Cyclic hydrocarbon group, 3-8 membered heterocyclic group, alkynyl group, C 6-10 Aryl or C 5-10 Substitution of heteroaryl groups, and / or optionally, the C 1-30 A C-shaped carbon atom is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 cycloalkyl, wherein formula (I) contains at least one of the C 1-30 Hydrocarbon functional group.

[0040] In some implementation schemes, R m and R n The connection forms a single bond.

[0041] In some implementation schemes, R m and R n The connection forms a single bond, and R1' and R B Each is H.

[0042] In some implementation schemes, R m and R n Each is independently H or C 1-6 Alkyl group, preferably H.

[0043] In some embodiments, formula (I) may have the structure of any one of formulas (Ia), (Ib), or (Ic):

[0044] The variables are defined as described in this paper, for example, as defined in equation (I).

[0045] In some embodiments, the hydrocarbon group or the C 1-30 Hydrocarbon groups can have 12-30 carbon atoms (C 12-30 Hydrocarbon group), 14-24 carbon atoms (C 14-24 (hydrocarbon group) or 16-22 carbon atoms (C 16-22 (hydrocarbon group) or 16-21 carbon atoms (C 16-21 Those with hydrocarbon groups.

[0046] In some embodiments, the hydrocarbon group or the C 1-30 The hydrocarbon group is optionally selected from hydroxyl, halogen, cyano, mercapto, azide, nitro, NR a R b C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 3-8 Cyclic hydrocarbon group, C 3-8 Cyclic hydrocarbon group, 3-8 membered heterocyclic group, C 6-10 Aryl or C 5-10 Substitution of heteroaryl groups, and / or optionally of the hydrocarbon group or the C 1-30 A C-shaped carbon atom is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 cycloalkyl, wherein R a and R b Each is independently H or C 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups.

[0047] In some embodiments, the hydrocarbon group or the C 1-30 The hydrocarbon group is optionally selected from hydroxyl, halogen, cyano, mercapto, azide, nitro, NR a R b C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 3-8 Cyclic hydrocarbon group, C 3-8 Substitution of the cyclic hydrocarbon group with an oxygen group, and / or optionally the hydrocarbon group or the C 1-30 A C-shaped carbon atom is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 cycloalkyl, wherein R a and R b Each is independently H or C1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups.

[0048] In some embodiments, the hydrocarbon group or the C 1-30 The hydrocarbon group is optionally selected from hydroxyl, halogen, cyano, mercapto, azide, nitro, NR a R b C 1-4 Haloalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy group substitution, and / or optionally the hydrocarbon group or the C 1-30 A C-shaped carbon atom is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 cycloalkyl, wherein R a and R b Each is independently H or C 1-4 Alkyl groups, such as methyl groups.

[0049] In some embodiments, the hydrocarbon group or the C 1-30 The hydrocarbon group is unsubstituted.

[0050] In some embodiments, the hydrocarbon group or the C 1-30 The hydrocarbon group is either fully saturated (e.g., as an alkyl group) or partially unsaturated (e.g., containing one or more carbon-carbon double bonds and / or carbon-carbon triple bonds, such as alkenyl or ynyl groups), preferably fully saturated.

[0051] In some embodiments, the hydrocarbon group or the C 1-30 The hydrocarbon group can be straight-chain or branched, preferably straight-chain.

[0052] In some embodiments, the hydrocarbon group or the C 1-30 The hydrocarbon group is connected to the rest of the molecule through a carbon atom at the 1-position or the last carbon atom.

[0053] In some embodiments, the hydrocarbon group includes alkyl, alkenyl, and alkynyl groups, optionally being straight-chain or branched. In some embodiments, the hydrocarbon group is C10. 1-30 Alkyl, such as C 12-30 Alkyl, C 14-24 Alkyl, C 16-22 Alkyl or C 16-21 Alkyl group, optionally straight-chain or branched. In some embodiments, the hydrocarbon group is C10. 2-30 alkenyl groups, such as C 12-30 alkenyl, C 14-24 alkenyl, C 16- 22 alkenyl or C 16-21 The alkenyl group, optionally, is straight-chain or branched. In some embodiments, the hydrocarbon group is C10.2-30 Alkyne groups, such as C 12-30 alkynyl group, C 14-24 alkynyl group, C 16-22 alkynyl or C 16-21 The alkynyl group, optionally the alkenyl group, is straight-chain or branched.

[0054] In some embodiments, the hydrocarbon group is C 16 C 17 C 18 C 19 C 20 C 21 Or C 22 Hydrocarbon groups, such as hexadecyl, hexadecylene, hexadecadienyl, heptadecanyl, heptadecanyl, heptadecanyl, octadecyl, octadecylene, octadecadienyl, octadectrienyl, nonadecanyl, nonadecanylene, nonadecanadienyl, nonadecantrienyl, eicosyl, eicosylene, eicosyladienyl, eicosyltrienyl, dodecyl, dodecylene, dodecylene, dodecylene, 6-octyltetradecyl, 10-hexylhexadecyl, all cis 7,10,13,16,19-docosapentaenyl, all cis 4,7,10,13,16,19-docosahexaenyl, all cis Formula 13,16-docosadienyl, all-cis-7,10,13,16-docosatetraenyl, all-cis-4,7,10,13,16-docosapentenyl or cis-13-docosaenyl, particularly 1-docoalkyl, 6-octyltetradecane-1-yl, 10-hexylhexadecane-1-yl, cis-docosa-13-en-1-yl, docosane-9-yl, docosane-2-yl, docosane-10-yl, docosane-11-yl or cis-4,7,10,13,16,19-docosahexaen-1-yl, wherein the hydrocarbon group may optionally be substituted with a group selected from the following: hydroxyl, amino, cyano, nitro, halogen, C 1-6 Alkoxy, C 3-8 Cyclic hydrocarbon group, C 3-8 Cycloalkyloxy group, 3-8 membered heterocyclic group, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, mercapto, azide, alkynyl, aryl, or heteroaryl, or optionally, a C10 group is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 Cycloalkyl.

[0055] In some embodiments, the hydrocarbon group is 1-hexadecyl, 1-eicosyl, 1-monodecyl, or 1-docodecyl.

[0056] In some implementations, formula (I) includes one of the C 1-30Hydrocarbon functional groups. In some embodiments, formula (I) comprises two of the C groups. 1-30 Hydrocarbon functional groups. In some embodiments, formula (I) comprises three of the C groups. 1-30 Hydrocarbon functional group.

[0057] In some embodiments, the present invention provides a nucleoside analog having the structure of formula (Ib) or a salt thereof:

[0058] in:

[0059] B is H or a modified or unmodified base;

[0060] R1 and R2 are independently H, hydroxyl protecting groups, reactive phosphorus groups, or solid supports;

[0061] X is selected from -O-, -S-, -SS-, -OC(=O), -C(=O)-O-, -NR4C(=O)-, -C(=O)NR4-, -NR4-, -O-NR4-, -NR4-O-, -O-NR4C(=O)-, or -C(=O)NR4-O-, where R4 is H or C. 1-6 Alkyl; and

[0062] R3 is C 1-30 Hydrocarbon groups, such as C 14-24 hydrocarbon group, C 16-22 The hydrocarbon group may optionally be substituted with a group selected from the following: hydroxyl, amino, cyano, nitro, halogen, C 1-6 Alkoxy, C 3-8 Cyclic hydrocarbon group, C 3-8 Cycloalkyloxy group, 3-8 membered heterocyclic group, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, mercapto, azide, alkynyl, aryl, or heteroaryl, or optionally, a C10 group is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 Cycloalkyl.

[0063] B

[0064] In some implementations, B is H, a base (including modified or unmodified bases), or -L. B -(C 1-30 (hydrocarbon group).

[0065] In some embodiments, B is H. In some embodiments, the backbone structure of the compound of the present invention wherein B is H is added to the 5' end and / or 3' end (preferably the 5' end or 3' end of the sense strand and / or antisense strand) of the dsRNA activator, or is used to replace the original nucleotide at an internal position of the sense strand and / or antisense strand.

[0066] In some embodiments, B is a base, such as a modified or unmodified (nuclear) base. In some embodiments, the skeletal structure of the compounds of the present invention in which B is a base (including a modified or unmodified base) is used to replace the original nucleotide at an internal position on the sense and / or antisense strands.

[0067] In some embodiments, B is an unmodified (nuclear)base, such as a native base. In other embodiments, B is a modified (nuclear)base, preferably any nucleobase capable of pairing with a native base without substantially affecting unwinding behavior, intracellular enzyme recognition, or the activity of the iRNA double strand.

[0068] In some implementations, B is a modified or unmodified purine or pyrimidine base.

[0069] In some embodiments, B is a modified or unmodified adenine base, guanine base, xanthine base, hypoxanthine base, cytosine base, thymine base, or uracil base, preferably a modified or unmodified adenine base, guanine base, cytosine base, thymine base, or uracil base.

[0070] In some implementations, B is adenine, guanine, cytosine, thymine, uracil, modified adenine, modified guanine, modified cytosine, modified thymine, or modified uracil.

[0071] Optionally, the modification includes, but is not limited to, substitution, denitrification, and hydrogenation. Optionally, for substitution modification, the substituent can be one or more functional groups or ligands, such as those selected from hydroxyl, halogen, cyano, azide, nitro, thio (=S) (e.g., carbonyl oxygen is substituted by a thio group), NR a N b 、(NR a N b )-C 1-6 Alkylene, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1- 6-alkoxy, C 1-6 Alkyl group, C 1-6 Alkyl-C(=S)-, C 6-10 Aryl, C 6-10 Aryl-C 1-6 Alkylene-, C 6-10 Aryl-C 1-6 imidene-, C 6-10 Aryl-C 1-6 Ethyne-, C5-10 heteroaryl, C 5-10 heteroaryl-C 1-6 Alkylene-, C 5-10 heteroaryl-C 1-6 imidene-, C 5-10 heteroaryl-C 1-6 α- and glycosyl groups, where R a and R b Each is independently H or C 1-6 Alkyl groups, preferably H or C 1-4 Alkyl groups, such as methyl groups.

[0072] Preferably, for substitution modification, the substituent can be selected from hydroxyl, halogen, cyano, azide, nitro, thio (=S), NR. a N b 、(NR a N b )-C 1-6 Alkylene, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkyl group, C 1-6 Alkyl-C(=S)-, C 6-10 Aryl, C 6-10 Aryl-C 1-6 Alkylene-, C 6-10 Aryl-C 1-6 imidene-, C 6-10 Aryl-C 1-6 Imyynyl-, where R a and R b Each is independently H or C 1-6 Alkyl groups, preferably H or C 1-4 Alkyl groups, such as methyl groups.

[0073] More preferably, for substitution modification, the substituent can be selected from hydroxyl, halogen, nitro, thio (=S), (NR) a N b )-C 1-6 Alkylene, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Hydroxyalkyl, C 1-6 Alkyl group, C 1-6 Alkyl-C(=S)-, C 6-10 Aryl, C 6-10 Aryl-C 1-6 Alkylene-, C 6-10 Aryl-C1-6 imidene-, C 6-10 Aryl-C 1-6 Imyynyl-, where R a and R b Each is independently H or C 1-6 Alkyl groups, preferably H or C 1-4 Alkyl groups, such as methyl groups.

[0074] More preferably, for substitution modification, the substituent can be selected from C 1-6 Alkyl group, C 1-6 Alkyl-C(=S)-, phenyl-C 1-4 alkylene-, phenyl-C 1-6 alkenyl-, phenyl-C 1-6 Isomerynyl-, such as C 1-6 Alkyl groups (such as butyryl) and phenyl-C 1-4 Alkylene (e.g., benzyl).

[0075] In some embodiments, B is a natural base, preferably adenine, guanine, cytosine, thymine, or uracil.

[0076] In some embodiments, B is a modified base, for example selected from 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxymethyl)uracil, 5-carboxymethylaminomethyl-2-thiouracil, 5-carboxymethylaminomethyluracil, dihydrouracil, β-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5 -Methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, β-D-mannosylQ nucleoside, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-hydroxyacetic acid (v), wybutoxosine, pseudouracil, Q nucleoside (queosine), 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-hydroxyacetic acid methyl ester, uracil-5-hydroxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl)uracil, (acp3)w, and 2,6-diaminopurine.

[0077] In some embodiments, B may be selected from 2,4-difluorotoluene, nitropyrrolyl, nitroindolyl, 8-aza-7-deadenine, 4-fluoro-6-methylbenzimidazole, 4-methylbenzimidazole, 3-methylisoquinolone, 5-methylisoquinolone, 3-methyl-7-propynylisoquinolone, 7-azaindolyl, 6-methyl-7-azaindolyl, imidazopyridyl, 9-methyl-imidazopyridyl, pyrrolopyrazinyl, isoquinolone, 7-propynylisoquinolone, propynyl-7-azaindolyl, 2,4,5-trimethylphenyl, 4-methylindolyl, 4,6-dimethylindolyl, phenyl, naphthyl, anthraceneyl, phenanthryl, pyreneyl, stilbene, tetraphenyl, pentaphenyl, and their structural derivatives.

[0078] In some embodiments, B is a modified or unmodified adenine base, guanine base, xanthine base, hypoxanthine base, cytosine base, thymine base, or uracil base, optionally the modification being performed by C. 1-6 Alkyl group, C 1-6 Alkyl-C(=S)-, phenyl-C 1-4 alkylene-, phenyl-C 1-6 alkenyl- or phenyl-C 1-6 Ethyne-substituted, for example, by C 1-6 Alkyl (such as butyryl) or phenyl-C 1-4 Alkylene-(e.g., benzyl) substitution.

[0079] In some embodiments, B is a modified or unmodified adenine base, guanine base, cytosine base, thymine base, or uracil base, optionally the modification being performed by C. 1-6 Alkyl group, C 1-6 Alkyl-C(=S)-, phenyl-C 1-4 alkylene-, phenyl-C 1-6 alkenyl- or phenyl-C 1-6 Ethyne-substituted, for example, by C 1-6 Alkyl (such as butyryl) or phenyl-C 1-4 Alkylene-(e.g., benzyl) substitution.

[0080] In some embodiments, B is a modified or unmodified adenine base, guanine base, cytosine base, thymine base, or uracil base, optionally the modification being performed by C. 1-6 Alkyl (such as butyryl) or benzyl substitution.

[0081] In some implementations, B is H or selected from the natural bases A, U, C, G, and T.

[0082] In some implementations, B is selected from non-natural purine bases.

[0083] In some implementations, B is selected from non-natural pyrimidine bases.

[0084] In some implementation schemes, B is selected from:

[0085] ◆Adenine, guanine, cytosine, thymine, uracil, xanthine, hypoxanthine;

[0086] ◆2-(halogenated)adenine, 2-(C 1-6 Alkyl)adenine, 2-(NR) a N b Adenine, 2-(aminoalkyl)adenine, 2-(methylthio)-N6-(isopentenyl)adenine, 6-(alkyl)adenine, 7-(denitro)adenine, 8-(halogenated)adenine, 8-(hydroxy)adenine, 8-(alkyl)adenine, 8-(alkenyl)adenine, 8-(alkynyl)adenine, 8-(amino)adenine, 8-(thioalkyl)adenine, 8-(thiol)adenine, N6-(alkyl)adenine, N6,N6-(dialkyl)adenine;

[0087] ◆2-(alkyl)guanine, 6-(alkyl)guanine, 7-(alkyl)guanine, 7-(denitro)guanine, 8-(alkyl)guanine, 8-(alkenyl)guanine, 8-(alkynyl)guanine, 8-(amino)guanine, 8-(halogenated)guanine, 8-(hydroxy)guanine, 8-(thioalkyl)guanine, 8-(thiol)guanine, N-(alkyl)guanine;

[0088] ◆2-(thio)cytosine, 3-(de-nitro)-5-(aza)cytosine, 3-(alkyl)cytosine, 5-(alkyl)cytosine, 5-(alkynyl)cytosine, 5-(halogenated)cytosine, 5-(trifluoromethyl)cytosine, 6-(azo)cytosine, N4-(acetyl)cytosine;

[0089] ◆3-(3-amino-3-carboxypropyl)uracil, 2-(thio)uracil, 5-(methyl)-2-(thio)uracil, 5-(methylaminomethyl)-2-(thio)uracil, 4-(thio)uracil, 5-(methyl)-4-(thio)uracil, 5-(methylaminomethyl)-4-(thio)uracil, 5-(methyl)-2,4-(dithio)uracil, 5-(methylaminomethyl)-2,4-(dithio)uracil, 5-(2-aminopropyl)uracil, 5-(alkyl)uracil, 5-(alkynyl)uracil, 5-(allylamino)uracil, 5-(aminoallyl)uracil, 5-(aminoalkyl)uracil, 5-(guanidinealkyl)uracil, 5-(1,3- Diazol-1-alkyl)uracil, 5-(cyanoalkyl)uracil, 5-(dialkylaminoalkyl)uracil, 5-(dimethylaminoalkyl)uracil, 5-(halogenated)uracil, 5-(methoxy)uracil, uracil-5-oxyacetic acid, 5-(methoxycarbonylmethyl)-2-(thio)uracil, 5-(methoxycarbonyl-methyl)uracil, 5-(propynyl)uracil, 5-(propynyl)uracil, 5-(trifluoromethyl)uracil, 6-(azo)uracil, dihydrouracil, N3-(methyl)uracil, 5-uracil (i.e., pseudouracil), 2-(thio)pseudouracil, 4-(thio)pseudouracil, 2,4-(dithio)pseudouracil, 5-(alkyl)pseudouracil 5-(methyl)pseudoruracil, 5-(alkyl)-2-(thio)pseudoruracil, 5-(methyl)-2-(thio)pseudoruracil, 5-(alkyl)-4-(thio)pseudoruracil, 5-(methyl)-4-(thio)pseudoruracil, 5-(alkyl)-2,4-(dithio)pseudoruracil, 5-(methyl)-2,4-(dithio)pseudoruracil, 1-substituted pseudouracil, 1-substituted 2-(thio)pseudoruracil, 1-substituted 4-(thio)pseudoruracil, 1-substituted 2,4-(dithio)pseudoruracil, 1-(aminocarbonylvinyl)-pseudoruracil, 1-(aminocarbonylvinyl)-2-(thio)pseudoruracil, 1-(aminocarbonylvinyl)-4-(thio)pseudoruracil Pyrimidine, 1-(aminocarbonylvinyl)-2,4-(dithio)pseudouracil, 1-(aminoalkylaminocarbonylvinyl)-pseudouracil, 1-(aminoalkylamino-carbonylvinyl)-2(thio)-pseudouracil, 1-(aminoalkylaminocarbonylvinyl)-4-(thio)pseudouracil, 1-(aminoalkylaminocarbonylvinyl)-2,4-(dithio)pseudouracil, 1,3-(diaza)-2-(oxo)-phenoxazin-1-yl, 1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl, 1,3-(diaza)-2-(oxo)-phenoxazin-1-yl, 1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl, 7-substituted 1,3-(diaza)-2-(oxo)-phenoxazine-1-yl, 7-substituted 1-(diaza)-2-(thio)-3-(diaza)-phenoxazine-1-yl, 7-substituted 1,3-(diaza)-2-(oxo)-phenoxazine-1-yl, 7-substituted 1-(diaza)-2-(thio)-3-(diaza)-phenoxazine-1-yl, 7-(aminoalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenoxazine-1-yl, 7-(aminoalkylhydroxy)-1-(diaza)-2-(thio)-3-(diaza)-phenoxazine-1-yl, 7-(aminoalkylhydroxy)-1,3-(diaza) -2-(oxo)-phenoxazine-1-yl, 7-(aminoalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazine-1-yl, 7-(guanidinealkylhydroxy)-1,3-(diaza)-2-(oxo)-phenoxazine-1-yl, 7-(guanidinealkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazine-1-yl, 7-(guanidinealkyl-hydroxy)-1,3-(diaza)-2-(oxo)-phenoxazine-1-yl, 7-(guanidinealkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazine-1-yl, 1,3,5-(triaza)-2 6-(dioxa)-naphthalene, inosine, xanthine, hypoxanthine, muscarin, tuberculin, isoguanosine, inosine, 2-aza-inosine, 7-deaza-inosine, nitroimidazolyl, nitropyrazolyl, nitrobenzimidazolyl, nitroindazolyl, aminoindole, pyrrolopyrimidinyl, 3-(methyl)isoquinolone, 5-(methyl)isoquinolone, 3-(methyl)-7-(propynyl)isoquinolone, 7-(aza)indole, 6-(methyl)-7-(aza)indole, imidazopyridyl, 9-(methyl)imidazopyridyl, pyrrolopyrazinyl, isoquinolone, 7-(propynyl)isoquinolone Propynyl-7-(aza)indolyl, 2,4,5-(trimethyl)phenyl, 4-(methyl)indolyl, 4,6-(dimethyl)indolyl, phenyl, naphthyl, anthraceneyl, phenanthryl, pyrene, stilbene, tetraphenyl, pentaphenyl, difluorotolyl, 4-(fluoro)-6-(methyl)benzimidazole, 4-(methyl)benzimidazole, 6-(azo)thymidine, 2-pyridone, 5-nitroindolyl, 3-nitropyrrole, 6-(aza)pyrimidine, 2-(amino)purine, 2,6-(diamino)purine, 5-substituted pyrimidine, N2-substituted purine, N6-substituted purine, O6-substituted purine, substituted 1,2,4-Triazole, pyrrolo-pyrimidin-2-one-3-yl, 6-phenyl-pyrrolo-pyrimidin-2-one-3-yl, para-substituted 6-phenyl-pyrrolo-pyrimidin-2-one-3-yl, ortho-substituted 6-phenyl-pyrrolo-pyrimidin-2-one-3-yl, di-ortho-substituted 6-phenyl-pyrrolo-pyrimidin-2-one-3-yl, p-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-one-3-yl, ortho-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-one-3-yl, di-ortho-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-one-3-yl, pyridinopyrimidin-3-yl, 2-oxo-7-amino-pyridinopyrimidin-3-yl, 2-oxo-pyridinopyrimidin-3-yl or any O-alkylated or N-alkylated derivative thereof.

[0090] In some implementations, B is independently selected from the following bases:

[0091] In some implementations, B is independently selected from the following bases:

[0092] In some implementations, B is -L B -(C 1-30 (Hydrocarbon group). In some embodiments, B is -L B -(C 1-30 The skeletal structure of the compound of the present invention (containing a hydrocarbon group) is added to the 5' end and / or 3' end (preferably the 5' end or 3' end of the sense strand) of the sense and / or antisense strands of the dsRNA activator. Optionally, the C 1-30 The hydrocarbon group is optionally substituted as described herein, and / or optionally, a C10 group is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 Cycloalkyl. Optionally, the C 1-30 The hydrocarbon group is further defined as in this article.

[0093] In some implementations, B is -L B -(C 1-30 Hydrocarbon group), preferably -L B -(C 12-30 (hydrocarbon group), more preferably -L B -(C 14-24 Hydrocarbon group), for example -L B -(C 16-22 (hydrocarbon group) or -L B -(C 20-22 (Hydrocarbon group). Optionally, the hydrocarbon group is fully saturated or partially unsaturated, and / or the hydrocarbon group is straight-chain or branched.

[0094] In some implementations, B is -L B -(C1-30 Alkyl groups, such as -L B -(C 12-30 alkyl), -L B -(C 14-24 alkyl), -L B -(C 16-22 alkyl) or -L B -(C 20-22 Alkyl group). Optionally, the alkyl group is straight-chain or branched.

[0095] In some implementations, B is -L B -(C 2-30 alkenyl), for example -L B -(C 12-30 alkenyl), -L B -(C 14-24 alkenyl), -L B -(C 16-22 alkenyl) or -L B -(C 20-22 Alkenyl group). Optionally, the alkenyl group is straight-chain or branched.

[0096] In some implementations, B is -L B -(C 2-30 (alkynyl group), such as -L B -(C 12-30 alkynyl group), -L B -(C 14-24 alkynyl group), -L B -(C 16-22 (alkynyl group) or -L B -(C 20-22 (Alkyne group). Optionally, the alkynyl group is straight-chain or branched.

[0097] In some implementations, L B This indicates non-existence, -O-, -S-, -C(O)-, -C(S)-, -N(R4)-, -C 1-16 Alkylene -, -(NHCH2CH2) m1 -、-(OCH2CH2) m2 -、-(SCH2CH2) m3 - A 4-8 membered subheterocyclic group or any combination of two or more thereof, wherein m1, m2, and m3 are each independently 1, 2, 3, 4, 5, or 6. Optionally, the 4-8 membered subheterocyclic group contains 1, 2, or 3 nitrogen atoms as heteroatoms.

[0098] In some implementations, L B The following characters represent non-existent elements: -O-, -S-, -C(O)-, -C(S)-, -N(R4)-, and -(NHCH2CH2).m1 -、-(NHCH2CH2) m1 -C(O)-、-(NHCH2CH2) m1 -C(S)-、-(OCH2CH2) m2 -、-(OCH2CH2) m2 -C(O)-、-(OCH2CH2) m2 -C(S)-、-(SCH2CH2) m3 -、-(SCH2CH2) m3 -C(O)-、-(SCH2CH2) m3 -C(S)-, 4-8 membered heterocyclic group, -N(R4)-C(O)-, -N(R4)-C(S)-, -N(R4)-C 1-16 Alkylene -C(O)-, -N(R4)-C 1-16 Alkylene-C(S)-, -N(R4)-C(O)-C 1-16 Alkylene-, -N(R4)-C(S)-C 1-16 Alkylene-,-N(R4)-C(O)-C 1- 16 Alkylene -C(O)-N(R4)-, -N(R4)-C(S)-C 1-16 Alkylene -C(O)-N(R4)-, -N(R4)-C(S)-C 1-16 Alkylene -C(S)-N(R4)-, -N(R4)-C(O)-C 1-16 Alkylene-C(S)-N(R4)-, -(4-8 membered heterocyclic)-C 1-16 Alkylene-(OCH2CH2) m2 -CO-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(SCH2CH2) m2 -CO-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(SCH2CH2) m2 -C(S)-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(OCH2CH2) m2 -C(S)-N(R4)-, wherein m1, m2, and m3 are each independently 1, 2, 3, 4, 5, or 6. Optionally, the 4-8 membered heterocyclic group is a 5-6 membered heteroaryl group, such as a triazolyl group.

[0099] In some implementations, L BThis indicates the absence of -O-, -S-, -C(O)-, -C(S)-, -N(R4)-, 4-8 membered subheterocyclic groups, -N(R4)-C(O)-, -N(R4)-C(S)-, -N(R4)-C(O)-C 1-16 Alkylene -C(O)-N(R4)-, -N(R4)-C(S)-C 1-16 Alkylene -C(O)-N(R4)-, -N(R4)-C(S)-C 1- 16 Alkylene -C(S)-N(R4)-, -N(R4)-C(O)-C 1-16 Alkylene-C(S)-N(R4)-, -(4-8 membered heterocyclic)-C 1-16 Alkylene-(OCH2CH2) m2 -CO-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(SCH2CH2) m2 -CO-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(SCH2CH2) m2 -C(S)-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(OCH2CH2) m2 -C(S)-N(R4)-, where m2 is 1, 2, 3, 4, 5, or 6. Optionally, the 4-8 membered subheterocyclic group contains 1, 2, or 3 nitrogen atoms as heteroatoms.

[0100] In some implementations, L B This represents -O-, 4-8 membered subheterocyclic groups, -N(R4)-C(O)-, -N(R4)-C(S)-, and -N(R4)-C(O)-C. 1-16 Alkylene -C(O)-N(R4)-, -N(R4)-C(S)-C 1-16 Alkylene -C(O)-N(R4)-, -N(R4)-C(S)-C 1-16 Alkylene -C(S)-N(R4)-, -N(R4)-C(O)-C 1-16 Alkylene-C(S)-N(R4)-, -(4-8 membered heterocyclic)-C 1-16 Alkylene-(OCH2CH2) m2 -CO-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(SCH2CH2) m2 -CO-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(SCH2CH2) m2-C(S)-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(OCH2CH2) m2 -C(S)-N(R4)-, where m2 is 1, 2, 3, 4, 5, or 6. Optionally, R4 is H or C. 1-4 Alkyl groups, such as methyl groups. Optionally, the 4-8 membered heterocyclic group contains 1, 2, or 3 nitrogen atoms as heteroatoms.

[0101] In some implementations, L B This represents -O-, 5-6-membered heteroaryl, -N(H)-C(O)-, -N(H)-C(S)-, -N(H)-C(O)-C 1-16 alkylene-C(O)-N(H)-, -(5-6-membered heteroaryl)-C 1-16 Alkylene-(OCH2CH2) m2 -CO-N(H)-, where m2 is 1, 2, 3, 4, 5, or 6. Optionally, the 5-6-membered heteroaryl group contains 1, 2, or 3 nitrogen atoms as heteroatoms, such as a triazolyl group.

[0102] In some implementations, L B This represents -O-, 5-6-membered heteroaryl, -N(H)-C(O)-, -N(H)-C(S)-, -N(H)-C(O)-C 1-12 alkylene-C(O)-N(H)-, -(5-6-membered heteroaryl)-C 1-6 Alkylene-(OCH2CH2) m2 -CO-N(H)-, where m2 is 1, 2, 3, 4, 5, or 6. Optionally, the 5-6-membered heteroaryl group contains 1, 2, or 3 nitrogen atoms as heteroatoms, such as a triazolyl group.

[0103] Understandable, L B The options can be connected in any direction, such as from left to right or from right to left.

[0104] In some implementations, B is selected from: H, adenine, guanine, cytosine, thymine, uracil, (For example ), -O-(C 12-30 Alkyl), -N(H)-C(O)-(C 12-30 alkyl), Optionally, the alkyl group is straight-chain or branched, preferably straight-chain.

[0105] In some implementations, B is selected from: adenine, guanine, cytosine, thymine, uracil, (For example ).

[0106] In some implementations, B or the -L B -(C 1-30 The hydrocarbon group is selected from: -O-(C 12-30 Alkyl), -N(H)-C(O)-(C 12-30 alkyl), Optionally, the alkyl group is straight-chain or branched, preferably straight-chain.

[0107] In some implementations, B or the -L B -(C 1-30 The hydrocarbon group is selected from: -O-(C 22 Straight-chain alkyl), -N(H)-C(O)-(C 21 (linear alkyl)

[0108] R1 and R2

[0109] In some embodiments, R1 and R2 are independently H, a hydroxyl protecting group, a reactive phosphorus group, or a C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 Hydrocarbon group.

[0110] In some embodiments, R1 and R2 are independently H, a hydroxyl protecting group, a reactive phosphorus group, or C. 1-30 Hydrocarbon group; optionally, R1 and R2 are not both C 1-30 Hydrocarbon group.

[0111] In some embodiments, one of R1 and R2 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 One component is a hydrocarbon group or solid support, and the other is a reactive phosphorus group. For example, in some embodiments, R1 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 A hydrocarbon group or solid support, and R2 is a reactive phosphorus group. In some embodiments, R1 is a reactive phosphorus group, and R2 is an H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or a C. 1-30 Hydrocarbon group or solid support. Preferably, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group. More preferably, R1 is a reactive phosphorus group, and R2 is a C24-hydroxyl protecting group. 1-30Hydrocarbon group. More preferably, R1 is a reactive phosphorus group, and R2 is a hydroxyl protecting group (such as DMTr). More preferably, both R1 and R2 are reactive phosphorus groups, and the reactive phosphorus groups may be the same or different. More preferably, R1 is a solid support, and R2 is a hydroxyl protecting group (such as DMTr).

[0112] In some embodiments, R1 is a hydroxyl protecting group; preferably, R1 is 4,4'-dimethoxytriphenylmethyl (DMTr).

[0113] In some embodiments, R2 is a reactive phosphorus group; preferably, R2 is (2-cyanoethyl-N,N-diisopropyl)phosphamide (-P-(N(i-Pr)2)O-CH2CH2-CN).

[0114] In some embodiments, the hydroxyl protecting group is selected from: trimethylsilyl (TMS), triethylsilyl (TES), dimethylisopropylsilyl (DMIPS), diethylisopropylsilyl (DEIPS), tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), triisopropylsilyl (TIPS), acetyl (Ac), chloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl (TFA), p-methoxybenzoyl, allyloxycarbonyl (Alloc), 2,2,2-trichloroethoxycarbonyl (Troc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), benzyl... The hydroxyl protecting group is 4,4'-dimethoxytriphenylmethyl (Bn), p-methoxybenzyl (PMB), allyl, triphenylmethyl (Tr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzylmethyl (PMBM), 4,4'-dimethoxytriphenylmethyl (DMTr), monomethoxytriphenylmethyl (MMT), 9-fluorenylmethoxycarbonyl (Fmoc), o-nitrophenylcarbonyl, p-phenylazophenylcarbonyl, benzoyl, p-chlorobenzoyl, and 5'-(α-methyl-2-nitropiperyl)oxycarbonyl (MeNPOC). Preferably, the hydroxyl protecting group is 4,4'-dimethoxytriphenylmethyl (DMTr).

[0115] In some embodiments, the reactive phosphorus group is derived from: a phosphate group (e.g., -P(=O)(OH)2), a thiophosphate group (e.g., -P(=S)(OH)2), an O,S-thiophosphate group (e.g., -P(=O)(OH)(SH)), a phosphite group (e.g., -P(OH)2), a phosphoramidite group, an activated phosphate group, an activated thiophosphate group, or an activated phosphite group.

[0116] In some embodiments, the reactive phosphorus group is derived from: an activated phosphate ester group, an activated phosphite ester group, or a phosphoramidite group.

[0117] In some embodiments, the reactive phosphorus group is derived from or is a phosphoramidite group, for example... Where R a and R b Each is independently H or C 1-6 Alkyl groups, and R c C is the optional replacement. 1-6 Alkyl groups, optionally substituted with hydroxyl, halogen, cyano, azide, NH2, or nitro groups. Preferably, R c It is a C that can be substituted with a cyano group. 1-6 Alkyl group. For example, the reactive phosphorus group is derived from (2-cyanoethyl-N,N-diisopropyl)phosphamide.

[0118] In some embodiments, the reactive phosphorus group is derived from a phosphate ester group, a thiophosphate ester group, or an O,S-thiophosphate ester group.

[0119] In some embodiments, the reactive phosphorus group is derived from either a phosphate ester group or a thiophosphate ester group.

[0120] In some implementations, the C 1-30 Hydrocarbon groups are as defined herein. For example, the C... 1-30 The hydrocarbon group is optionally substituted as described herein, and / or optionally, a C10 group is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 Cycloalkyl.

[0121] In some implementations, the C 1-30 The hydrocarbon group is C 12-30 Hydrocarbon group, preferably C 14-24 Hydrocarbon group, more preferably C 16-22 The hydrocarbon group is fully saturated or partially unsaturated, and / or the hydrocarbon group is straight-chain or branched. In some embodiments, the C 1-30 Hydrocarbon groups include C 1-30 Alkyl, C 2- 30 alkenyl and C 2-30 The alkynyl group, optionally the alkyl, alkenyl, or alkynyl group, is straight-chain or branched. In some embodiments, the hydrocarbon group is C10. 1-30 Alkyl, such as C 12-30 Alkyl, C 14-24 Alkyl or C 16-22 Alkyl group, optionally straight-chain or branched. In some embodiments, the hydrocarbon group is C10. 2-30 alkenyl groups, such as C 12-30alkenyl, C 14-24 alkenyl or C 16-22 The alkenyl group, optionally, is straight-chain or branched. In some embodiments, the alkynyl group is C0. 2-30 Alkyne groups, such as C 12-30 alkynyl group, C 14-24 alkynyl or C 16-22 The alkynyl group, optionally, is straight-chain or branched.

[0122] In some embodiments, the solid support is a resin, for example...

[0123] In some embodiments, R1 is a hydroxyl protecting group such as DMTr, and R2 is a phosphoramidite group, such as... Where R a and R b Each is independently H or C 1-6 Alkyl groups, and R c It is a C group that is optionally substituted with a hydroxyl, halogen, cyano, azide, NH2, or nitro group. 1-6 Alkyl groups, preferably C groups optionally substituted with cyano groups. 1-6 Alkyl group. Preferably, R1 is a hydroxyl protecting group such as DMTr, and R2 is...

[0124] In some embodiments, R1 is a phosphoramidite group, such as Where R a and R b Each is independently H or C 1-6 Alkyl groups, and R c It is a C group that is optionally substituted with a hydroxyl, halogen, cyano, azide, NH2, or nitro group. 1-6 Alkyl groups, preferably C groups optionally substituted with cyano groups. 1-6 Alkyl group, and R2 is C 1-30 Hydrocarbon group, optionally the C 1-30 The hydrocarbon group is as defined herein. Preferably, R1 is... R2 is C 1-30 Alkyl, such as C 12-30 Alkyl, C 14-24 Alkyl or C 16-22 Alkyl groups, optionally, are straight-chain or branched, preferably straight-chain.

[0125] In some embodiments, R1 is a phosphoramidite group, such as Where R a and R b Each is independently H or C 1-6 Alkyl groups, and R c It is a C group that is optionally substituted with a hydroxyl, halogen, cyano, azide, NH2, or nitro group. 1-6Alkyl groups, preferably C groups optionally substituted with cyano groups. 1-6 Alkyl group, and R2 is a hydroxyl protecting group such as DMTr. Preferably, R1 is... R2 and hydroxyl protecting groups, such as DMTr.

[0126] In some embodiments, both R1 and R2 are phosphoramidyl groups, such as Where R a and R b Each is independently H or C 1-6 Alkyl groups, and R c It is a C group that is optionally substituted with a hydroxyl, halogen, cyano, azide, NH2, or nitro group. 1-6 Alkyl groups, preferably C groups optionally substituted with cyano groups. 1-6 Alkyl groups, and R2 is a hydroxyl protecting group such as DMTr. Preferably, both R1 and R2 are alkyl groups.

[0127] In some implementations, R1 is a solid support such as R2 and hydroxyl protecting groups, such as DMTr.

[0128] XR3 and R B

[0129] In some implementations, X is selected from non-existent, -O-, -S-, -SS-, -OC(=O), -C(=O)-O-, -NR4C(=O)-, -C(=O)NR4-, -NR4-, -O-NR4-, -NR4-O-, -O-NR4-C(O)-, and -C(=O)NR4-O-, where R4 is H or C. 1-6 Alkyl groups, preferably H or C 1-4 Alkyl groups, such as methyl groups.

[0130] In some implementations, X is selected from non-existent, -O-, -S-, -O-NR4-, -NR4-O-, -O-NR4-C(O)-, and -C(=O)NR4-O-, where R4 is H or C. 1-6 Alkyl groups, preferably H or C 1-4 Alkyl groups, such as methyl groups.

[0131] In some implementations, X is selected from non-existent, -O-, -O-NR4-, -NR4-O-, and -O-NR4-C(O)-, where R4 is H or C. 1-6 Alkyl groups, preferably H or C 1-4 Alkyl groups, such as methyl groups.

[0132] In some implementations, X is absent.

[0133] In some embodiments, X is selected from -O-, -O-NR4-, -NR4-O-, and -O-NR4C(=O)-, where R4 is H or methyl.

[0134] In some implementations, X is selected from *-O-NR4-, *-NR4-O-, and *-O-NR4-CO-, where R4 is H or C. 1-6 Alkyl groups, preferably H or C 1-4 Alkyl groups, such as methyl groups, and wherein the *-labeled valence bond is connected to a pentose ring. Preferably, X is selected from *-ON(CH3)-, *-N(CH3)-O-, *-O-NH-CO-, and *-ON(CH3)-CO-, wherein the *-labeled valence bond is connected to a pentose ring.

[0135] In some implementations, R3 is H or C. 1-30 A hydrocarbon group, wherein the hydrocarbon group is optionally substituted as described herein, and / or optionally a C10 group is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 Cycloalkyl.

[0136] In some implementations, R3 is H.

[0137] In some implementations, R3 is C 1-6 A hydrocarbon group, wherein the hydrocarbon group is fully saturated or partially unsaturated. In some embodiments, R3 is C 1- 6-alkyl (such as methyl), C 2-6 alkenyl or C 2-6 Alkyne group.

[0138] In some implementations, R3 is C 1-30 Hydrocarbon group, preferably C 12-30 hydrocarbon group, C 14-24 hydrocarbon group, C 16-22 hydrocarbon group or C 16-21 The hydrocarbon group is fully saturated or partially unsaturated, and / or the hydrocarbon group is straight-chain or branched. In some embodiments, the C 1-30 Hydrocarbon groups include C 1-30 Alkyl, C 2-30 alkenyl and C 2-30 The alkynyl group, optionally the alkyl, alkenyl, or alkynyl group, is straight-chain or branched. In some embodiments, the hydrocarbon group is C10. 1-30 Alkyl, such as C 12-30 Alkyl, C 14-24 Alkyl, C 16-22 Alkyl or C 16-21 Alkyl group, optionally straight-chain or branched. In some embodiments, the hydrocarbon group is C10. 2-30 alkenyl groups, such as C 12-30 alkenyl, C 14-24 alkenyl, C16-22 alkenyl or C 16-21 The alkenyl group, optionally, is straight-chain or branched. In some embodiments, the alkynyl group is C0. 2-30 Alkyne groups, such as C 12-30 alkynyl group, C 14-24 alkynyl group, C 16-22 alkynyl or C 16- 21 The alkynyl group, optionally, is straight-chain or branched.

[0139] In some implementations, R3 is C 16-22 Hydrocarbon groups, such as C 16 C 17 C 18 C 19 C 20 C 21 Or C 22 Hydrocarbon groups, such as hexadecyl, hexadecylene, hexadecadienyl, heptadecanyl, heptadecanyl, heptadecanyl, octadecyl, octadecylene, octadecadienyl, octadectrienyl, nonadecanyl, nonadecanylene, nonadecanadienyl, nonadecantrienyl, eicosyl, eicosylene, eicosyladienyl, eicosyltrienyl, dodecyl, dodecylene, dodecylene, dodecylene, 6-octyltetradecyl, 10-hexylhexadecyl, all cis 7,10,13,16,19-docosapentaenyl, all cis 4,7,10,13,16,19-docosahexaenyl, all cis Formula 13,16-docosadienyl, all-cis-7,10,13,16-docosatetraenyl, all-cis-4,7,10,13,16-docosapentenyl or cis-13-docosaenyl, particularly 1-docoalkyl, 6-octyltetradecane-1-yl, 10-hexylhexadecane-1-yl, cis-docosa-13-en-1-yl, docosane-9-yl, docosane-2-yl, docosane-10-yl, docosane-11-yl or cis-4,7,10,13,16,19-docosahexaen-1-yl, wherein the hydrocarbon group may optionally be substituted with a group selected from the following: hydroxyl, amino, cyano, nitro, halogen, C 1-6 Alkoxy, C 3-8 Cyclic hydrocarbon group, C 3-8 Cycloalkyloxy group, 3-8 membered heterocyclic group, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, mercapto, azide, alkynyl, aryl, or heteroaryl, or optionally, a C10 group is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 Cycloalkyl.

[0140] In some embodiments, R3 is 1-hexadecyl, 1-eicosyl, 1-monodecyl, or 1-docodecyl.

[0141] In some implementation schemes, R B For H or C 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups.

[0142] In some implementation schemes, R B For H.

[0143] In some implementation schemes, R B Together with XR3, they form C 2-5 The alkylene chain, such as a C3 or C4 alkylene chain, is optionally substituted by one or more groups independently selected from the following: hydroxyl, halogen, cyano, mercapto, azide, nitro, NR. a R b C 1- 6-alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 1-6 alkoxy group, where R a and R b Each is independently H or C 1-6 Alkyl, preferably C 1-4 Alkyl groups, such as methyl groups.

[0144] In some implementation schemes, R B Together with XR3, they form C 2-5 Alkylene chains, such as C3 alkylene chains or C4 alkylene chains.

[0145] In some implementations, (I) may have the structure of formula (Id):

[0146] The variables are as defined in this paper.

[0147] R1'

[0148] In some implementations, R1' is H or -C 0-6 alkylene-O-(C 1-30 hydrocarbon group), -C 0-6 alkylene-S-(C 1-30 hydrocarbon group), -C 0-6 Alkylene-NR4-(C 1-30 hydrocarbon group), -C 0-6 Alkylene-C(O)-(C 1-30 hydrocarbon group), -C 0-6 Alkylene-C(S)-(C1-30 (hydrocarbon group), wherein the C 1-30 Hydrocarbon groups are as defined in this article.

[0149] In some implementations, R1' is H or -C 0-6 alkylene-O-(C 1-30 (hydrocarbon group) or -C 0-6 alkylene-S-(C 1-30 Hydrocarbon group), preferably H or -C 0-6 alkylene-O-(C 1-30 Hydrocarbon group), more preferably H or -C 1-6 alkylene-O-(C 1-30 (hydrocarbon group), wherein the C 1-30 Hydrocarbon groups are as defined in this article.

[0150] In some implementations, R1' is H.

[0151] In some implementations, R1' is -C 0-6 alkylene-O-(C 1-30 Hydrocarbon group), preferably -C 1-6 alkylene-O-(C 1-30 (hydrocarbon group).

[0152] Optionally, the C 1-30 The hydrocarbon group is C 12-30 Hydrocarbon group, preferably C 14-24 Hydrocarbon group, more preferably C 16-22 hydrocarbon group or C 16-21 The hydrocarbon group is fully saturated or partially unsaturated, and / or the hydrocarbon group is straight-chain or branched. In some embodiments, the C 1-30 Hydrocarbon groups include C 1-30 Alkyl, C 2-30 alkenyl and C 2-30 The alkynyl group, optionally the alkyl, alkenyl, or alkynyl group, is straight-chain or branched. In some embodiments, the hydrocarbon group is C10. 1-30 Alkyl, such as C 12-30 Alkyl, C 14-24 Alkyl, C 16-22 Alkyl or C 16-21 Alkyl group, optionally straight-chain or branched. In some embodiments, the hydrocarbon group is C10. 2-30 alkenyl groups, such as C 12-30 alkenyl, C 14-24 alkenyl, C 16-22 alkenyl or C 16-21 The alkenyl group, optionally, is straight-chain or branched. In some embodiments, the alkynyl group is C0. 2-30 Alkyne groups, such as C 12-30 alkynyl group, C 14-24 alkynyl group, C16- 22 alkynyl or C 16-21 The alkynyl group, optionally, is straight-chain or branched.

[0153] In some implementations, R1' is -C 0-6 Alkylene -O-, -C 0-6 Alkylene-S-, -C 0-6 Alkylene-NR4-, -C 0-6 Alkylene -C(O)-, -C 0- 6-alkylene-C(S)-, preferably -O-, -S-, -NR4-, -C(O)-, -C(S)-, more preferably -C(O)- or -C(S)-, XR3 indicates Furthermore, R1' connects with XR3 to form a bridge ring, where the wavy line indicates that the valence bond is connected to R1', and the asterisk indicates that the valence bond is connected to the rest of the molecule.

[0154] In some implementations, R1' is -C(O)-, and XR3 represents... Furthermore, R1' connects to XR3 to form a bridging ring, where the wavy line indicates the valence bond connecting to R1', and the asterisk indicates the valence bond connecting to the rest of the molecule. For example, in some embodiments, formula (I) has the structure shown in formula (Ie):

[0155] In some embodiments, a compound of formula (Ia) or a salt thereof (e.g., a pharmaceutically acceptable salt), a solvate (e.g., a hydrate), or an isotopically labeled compound (e.g., a deuterated compound) is provided:

[0156] in:

[0157] B is H, a base (including modified or unmodified bases), or -L. B -(C 1-30 (hydrocarbon group), of which L B It is a linker; preferably, B is H or a base (including modified or unmodified bases);

[0158] R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 hydrocarbon group;

[0159] X is selected from non-existent, -O-, -O-NR4-, -NR4-O-, and -O-NR4-C(O)-;

[0160] R3 is H or C. 1-30 Hydrocarbon group, preferably C 1-30hydrocarbon group;

[0161] R B For H or C 1-6 Alkyl groups, preferably H;

[0162] R1' is H or -C 0-6 alkylene-O-(C 1-30 Hydrocarbon group), preferably H;

[0163] R4 is H or C. 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups;

[0164] Wherein C 1-30 The hydrocarbon group is fully saturated or partially unsaturated, and wherein formula (Ia) contains at least one of the C groups. 1-30 Hydrocarbon functional groups. Optionally, the variables are further defined as herein. Optionally, the C 1-30 The hydrocarbon group is C 12-30 hydrocarbon group, C 14-24 hydrocarbon group or C 16-22 The hydrocarbon group is fully saturated or partially unsaturated, and / or the hydrocarbon group is straight-chain or branched. Optionally, the hydrocarbon group is C24. 1-30 Alkyl, such as C 12-30 Alkyl, C 14-24 Alkyl or C 16-22 Alkyl group, optionally straight-chain or branched. Optionally, the hydrocarbon group is C10. 2-30 alkenyl groups, such as C 12-30 alkenyl, C 14-24 alkenyl or C 16-22 Alkenyl group, optionally the alkenyl group is straight-chain or branched. Optionally, the alkynyl group is C10. 2- 30 Alkyne groups, such as C 12-30 alkynyl group, C 14-24 alkynyl or C 16-22 The alkynyl group is optionally straight-chain or branched. Optionally, the reactive phosphorus group is derived from a phosphate ester group, a thiophosphate ester group, an O,S-thiophosphate ester group, or a phosphoramidite group.

[0165] In some embodiments, R1 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 A hydrocarbon group or solid support, and R2 is a reactive phosphorus group. In some embodiments, R1 is a reactive phosphorus group, and R2 is an H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or a C. 1-30 Hydrocarbon group or solid support. Preferably, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group. More preferably, R1 is a reactive phosphorus group, and R2 is a C24-hydroxyl protecting group. 1-30Hydrocarbon group. More preferably, R1 is a reactive phosphorus group, and R2 is a hydroxyl protecting group (such as DMTr). More preferably, both R1 and R2 are reactive phosphorus groups, and the reactive phosphorus groups may be the same or different. More preferably, R1 is a solid support, and R2 is a hydroxyl protecting group (such as DMTr).

[0166] In some embodiments, a compound of formula (Ia) or a salt thereof (e.g., a pharmaceutically acceptable salt), a solvate (e.g., a hydrate), or an isotopically labeled compound (e.g., a deuterated compound) is provided, wherein B is H. The backbone structure of the compound of formula (Ia) wherein B is H can be added to the 5' end and / or 3' end (preferably the 5' end or 3' end of the sense strand) of the dsRNA activator, or can be used to replace the pronucleotide at an internal position of the sense strand and / or antisense strand.

[0167] In some embodiments, a compound of formula (Ia) or a salt thereof (e.g., a pharmaceutically acceptable salt), a solvate (e.g., a hydrate), or an isotope-labeled compound (e.g., a deuterated compound) is provided, wherein:

[0168] B is H;

[0169] R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 hydrocarbon group;

[0170] X is non-existent, -O-, -O-NR4-, -NR4-O-, or -O-NR4-C(O)-; preferably, it is non-existent, -O-, -O-NR4-, or -NR4-O-;

[0171] R3 is H or C. 1-30 Hydrocarbon group, preferably C 1-30 hydrocarbon group;

[0172] R B For H or C 1-6 Alkyl groups, preferably H;

[0173] R1' is H or -C 0-6 alkylene-O-(C 1-30 Hydrocarbon group), preferably H;

[0174] R4 is H or C. 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups;

[0175] Wherein C 1-30 The hydrocarbon group is fully saturated or partially unsaturated, and wherein formula (Ia) contains at least one of the C groups. 1-30Hydrocarbon functional groups. Optionally, the variables are further defined as described herein, for example, as defined above.

[0176] In some embodiments, a compound of formula (Ia) or a salt (e.g., a pharmaceutically acceptable salt), a solvate (e.g., a hydrate), or an isotopically labeled compound (e.g., a deuterated compound) is provided, wherein B is a base (including modified or unmodified bases). The skeletal structure of the compound of formula (Ia), wherein B is a base, can be used to replace the original nucleotide at an internal position of the sense and / or antisense strands.

[0177] In some embodiments, a compound of formula (Ia) or a salt thereof (e.g., a pharmaceutically acceptable salt), a solvate (e.g., a hydrate), or an isotope-labeled compound (e.g., a deuterated compound) is provided, wherein:

[0178] B is a base (including modified or unmodified bases);

[0179] R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 Hydrocarbon group; preferably, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group;

[0180] X is selected from -O-, -O-NR4-, -NR4-O-, and -O-NR4-C(O)-;

[0181] R3 is H or C. 1-30 Hydrocarbon group, preferably C 1-30 hydrocarbon group;

[0182] R B For H or C 1-6 Alkyl groups, preferably H;

[0183] R1' is H or -C 0-6 alkylene-O-(C 1-30 Hydrocarbon group), preferably H;

[0184] R4 is H or C. 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups;

[0185] Wherein C 1-30 The hydrocarbon group is fully saturated or partially unsaturated, and wherein formula (Ia) contains at least one of the C groups. 1-30 Hydrocarbon functional groups. Optionally, the variables are further defined as described herein, for example, as defined above.

[0186] In some embodiments, a compound of formula (Ia) or a salt thereof (e.g., a pharmaceutically acceptable salt), a solvate (e.g., a hydrate), or an isotope-labeled compound (e.g., a deuterated compound) is provided, wherein:

[0187] B is H or a base (including modified or unmodified bases), preferably a base (including modified or unmodified bases);

[0188] R1 and R2 are independently H, a hydroxyl protecting group, a reactive phosphorus group, or a C. 1-30 Hydrocarbon group; optionally, R1 and R2 are not both C 1-30 Hydrocarbon group; preferably, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group;

[0189] X is selected from -O-;

[0190] R3 is C 1-6 A hydrocarbon group, wherein the hydrocarbon group is fully saturated or partially unsaturated;

[0191] R B For H or C 1-6 Alkyl groups, preferably H;

[0192] R1' is -C 0-6 alkylene-O-(C 1-30 Hydrocarbon group), preferably -C 1-6 alkylene-O-(C 1-30 (hydrocarbon group);

[0193] Wherein C 1-30 The hydrocarbon groups are either fully saturated or partially unsaturated. Optionally, the variables are further defined as described herein, for example, as defined above.

[0194] In some embodiments, a compound of formula (Ib) or a salt thereof (e.g., a pharmaceutically acceptable salt), a solvate (e.g., a hydrate), or an isotopically labeled compound (e.g., a deuterated compound) is provided:

[0195] in:

[0196] B is H, a base (including modified or unmodified bases), or -L. B -(C 1-30 (hydrocarbon group), of which L B It is a linker; preferably, B is H or a modified or unmodified base;

[0197] R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 hydrocarbon group

[0198] X is selected from non-existent, -O-, -O-NR4-, -NR4-O-, and -O-NR4-C(O)-;

[0199] R3 is H or C. 1-30 Hydrocarbon group, preferably C 1-30 hydrocarbon group;

[0200] R4 is H or C. 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups;

[0201] Wherein C 1-30 The hydrocarbon group is fully saturated or partially unsaturated, and wherein formula (Ib) contains at least one of the C groups. 1-30 Hydrocarbon functional groups. Optionally, the variables are further defined as herein. Optionally, the C 1-30 The hydrocarbon group is C 12-30 hydrocarbon group, C 14-24 hydrocarbon group or C 16-22 The hydrocarbon group is fully saturated or partially unsaturated, and / or the hydrocarbon group is straight-chain or branched. Optionally, the hydrocarbon group is C24. 1-30 Alkyl, such as C 12-30 Alkyl, C 14-24 Alkyl or C 16-22 Alkyl group, optionally straight-chain or branched. Optionally, the hydrocarbon group is C10. 2-30 alkenyl groups, such as C 12-30 alkenyl, C 14-24 alkenyl or C 16-22 Alkenyl group, optionally the alkenyl group is straight-chain or branched. Optionally, the alkynyl group is C10. 2- 30 Alkyne groups, such as C 12-30 alkynyl group, C 14-24 alkynyl or C 16-22 The alkynyl group is optionally straight-chain or branched. Optionally, the reactive phosphorus group is derived from a phosphate ester group, a thiophosphate ester group, an O,S-thiophosphate ester group, or a phosphoramidite group.

[0202] In some embodiments, R1 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 A hydrocarbon group or solid support, and R2 is a reactive phosphorus group. In some embodiments, R1 is a reactive phosphorus group, and R2 is an H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or a C. 1-30 Hydrocarbon group or solid support. Preferably, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group. More preferably, R1 is a reactive phosphorus group, and R2 is a C24-hydroxyl protecting group. 1-30Hydrocarbon group. More preferably, R1 is a reactive phosphorus group, and R2 is a hydroxyl protecting group (such as DMTr). More preferably, both R1 and R2 are reactive phosphorus groups, and the reactive phosphorus groups may be the same or different. More preferably, R1 is a solid support, and R2 is a hydroxyl protecting group (such as DMTr).

[0203] In some embodiments, a compound of formula (Ib) or a salt (e.g., a pharmaceutically acceptable salt), a solvate (e.g., a hydrate), or an isotopically labeled compound (e.g., a deuterated compound) is provided, wherein B is H. The backbone structure of the compound of formula (Ib) wherein B is H can be added to the 5' end and / or 3' end (preferably the 5' end or 3' end of the sense strand) of the dsRNA activator, or can be used to replace the pronucleotide at an internal position of the sense strand and / or antisense strand.

[0204] In some embodiments, a compound of formula (Ib) or a salt thereof (e.g., a pharmaceutically acceptable salt), a solvate (e.g., a hydrate), or an isotope-labeled compound (e.g., a deuterated compound) is provided, wherein:

[0205] B is H;

[0206] R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 hydrocarbon group;

[0207] X is non-existent, -O-, -O-NR4-, -NR4-O-, or -O-NR4-C(O)-; preferably, it is non-existent, -O-, -O-NR4-, or -NR4-O-;

[0208] R3 is H or C. 1-30 Hydrocarbon group, preferably C 1-30 hydrocarbon group;

[0209] R4 is H or C. 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups;

[0210] Wherein C 1-30 The hydrocarbon group is fully saturated or partially unsaturated, and wherein formula (Ib) contains at least one of the C groups. 1-30 Hydrocarbon functional groups. Optionally, the variables are further defined as described herein, for example, as defined above.

[0211] In some embodiments, a compound of formula (Ib) or a salt (e.g., a pharmaceutically acceptable salt), a solvate (e.g., a hydrate), or an isotopically labeled compound (e.g., a deuterated compound) is provided, wherein B is a base (including modified or unmodified bases). The skeletal structure of the compound of formula (Ib) wherein B is a base can be used to replace the pronucleotide at an internal position of the sense and / or antisense strands.

[0212] In some embodiments, a compound of formula (Ib) or a salt thereof (e.g., a pharmaceutically acceptable salt), a solvate (e.g., a hydrate), or an isotope-labeled compound (e.g., a deuterated compound) is provided, wherein:

[0213] B is a base (including modified or unmodified bases);

[0214] R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 Hydrocarbon group; preferably, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group;

[0215] X is selected from -O-, -O-NR4-, -NR4-O-, and -O-NR4-C(O)-;

[0216] R3 is H or C. 1-30 Hydrocarbon group, preferably C 1-30 hydrocarbon group;

[0217] R4 is H or C. 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups;

[0218] Wherein C 1-30 The hydrocarbon group is fully saturated or partially unsaturated, and wherein formula (Ib) contains at least one of the C groups. 1-30 Hydrocarbon functional groups. Optionally, the variables are further defined as described herein, for example, as defined above.

[0219] Optionally, the skeletal structure of the compound of formula (Ib), where B is a base (including modified or unmodified bases), is used to replace the original nucleotide at an internal position of the sense and / or antisense strands.

[0220] In some embodiments, a compound of formula (Ic) or a salt thereof (e.g., a pharmaceutically acceptable salt), a solvate (e.g., a hydrate), or an isotopically labeled compound (e.g., a deuterated compound) is provided:

[0221] in:

[0222] B is H or a base (including modified or unmodified bases), preferably a base (including modified or unmodified bases);

[0223] R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 hydrocarbon group;

[0224] X is selected from non-existent, -O-, -O-NR4-, -NR4-O-, and -O-NR4-C(O)-, for example, -NR4-O-;

[0225] R3 is H or C. 1-30 Hydrocarbon group, preferably C 1-30 hydrocarbon group;

[0226] R B For H or C 1-6 Alkyl groups, preferably H;

[0227] R1' is H;

[0228] Wherein C 1-30 The hydrocarbon group is fully saturated or partially unsaturated, and wherein formula (Ic) contains at least one of the C groups. 1-30 Hydrocarbon functional groups. Optionally, the variables are further defined as herein. Optionally, the C 1-30 The hydrocarbon group is C 12-30 hydrocarbon group, C 14-24 hydrocarbon group or C 16-22 The hydrocarbon group is fully saturated or partially unsaturated, and / or the hydrocarbon group is straight-chain or branched. Optionally, the hydrocarbon group is C24. 1-30 Alkyl, such as C 12-30 Alkyl, C 14-24 Alkyl or C 16-22 Alkyl group, optionally straight-chain or branched. Optionally, the hydrocarbon group is C10. 2-30 alkenyl groups, such as C 12-30 alkenyl, C 14-24 alkenyl or C 16-22 Alkenyl group, optionally the alkenyl group is straight-chain or branched. Optionally, the alkynyl group is C10. 2- 30 Alkyne groups, such as C 12-30 alkynyl group, C 14-24 alkynyl or C 16-22 The alkynyl group is optionally straight-chain or branched. Optionally, the reactive phosphorus group is derived from a phosphate ester group, a thiophosphate ester group, an O,S-thiophosphate ester group, or a phosphoramidite group.

[0229] In some embodiments, R1 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 A hydrocarbon group or solid support, and R2 is a reactive phosphorus group. In some embodiments, R1 is a reactive phosphorus group, and R2 is an H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or a C. 1-30 Hydrocarbon group or solid support. Preferably, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group. More preferably, R1 is a reactive phosphorus group, and R2 is a C24-hydroxyl protecting group. 1-30 Hydrocarbon group. More preferably, R1 is a reactive phosphorus group, and R2 is a hydroxyl protecting group (such as DMTr). More preferably, both R1 and R2 are reactive phosphorus groups, and the reactive phosphorus groups may be the same or different. More preferably, R1 is a solid support, and R2 is a hydroxyl protecting group (such as DMTr).

[0230] Optionally, the skeletal structure of the compound of formula (Ic), where B is a base (including modified or unmodified bases), is used to replace the original nucleotide at an internal position of the sense and / or antisense strands.

[0231] In some embodiments, a compound of formula (Id) or a salt thereof (e.g., a pharmaceutically acceptable salt), a solvate (e.g., a hydrate), or an isotopically labeled compound (e.g., a deuterated compound) is provided:

[0232] in:

[0233] B is -L B -(C 1-30 (hydrocarbon group), of which L B It is a linker;

[0234] R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 hydrocarbon group;

[0235] R1' is H or -C 0-6 alkylene-O-(C 1-30 Hydrocarbon group), preferably H;

[0236] Wherein C 1-30 The hydrocarbon group may be fully saturated or partially unsaturated. Optionally, the variables are as defined herein. Optionally, the C 1-30 The hydrocarbon group is C 12-30 hydrocarbon group, C 14-24 hydrocarbon group or C 16-22 The hydrocarbon group is fully saturated or partially unsaturated, and / or the hydrocarbon group is straight-chain or branched; optionally, the hydrocarbon group is C 1-30 Alkyl, such as C12-30 Alkyl, C 14-24 Alkyl or C 16-22 Alkyl group, optionally the alkyl group is straight-chain or branched; optionally, the hydrocarbon group is C10. 2-30 alkenyl groups, such as C 12-30 alkenyl, C 14-24 alkenyl or C 16-22 Alkenyl group, optionally the alkenyl group is straight-chain or branched; optionally, the alkynyl group is C 2-30 Alkyne groups, such as C 12-30 alkynyl group, C 14-24 alkynyl or C 16-22 The alkynyl group, optionally, is straight-chain or branched.

[0237] Optional, L B This represents -O-, 5-6-membered heteroaryl, -N(H)-C(O)-, -N(H)-C(S)-, -N(H)-C(O)-C 1-16 alkylene-C(O)-N(H)-, -(5-6-membered heteroaryl)-C 1-16 Alkylene-(OCH2CH2) m2 -CO-N(H)-, where m2 is 1, 2, 3, 4, 5, or 6. Preferably, L B This represents -O-, 5-6-membered heteroaryl, -N(H)-C(O)-, -N(H)-C(S)-, -N(H)-C(O)-C 1-12 alkylene-C(O)-N(H)-, -(5-6-membered heteroaryl)-C 1-6 Alkylene-(OCH2CH2) m2 -CO-N(H)-, where m2 is 1, 2, 3, 4, 5, or 6. Optionally, the 5-6-membered heteroaryl group contains 1, 2, or 3 nitrogen atoms as heteroatoms, such as a triazolyl group.

[0238] Optionally, the reactive phosphorus group is derived from a phosphate ester group, a thiophosphate ester group, an O,S-thiophosphate ester group, or a phosphoramidite group.

[0239] Optionally, the solid support is a resin, for example...

[0240] In some embodiments, R1 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 A hydrocarbon group or solid support, and R2 is a reactive phosphorus group. In some embodiments, R1 is a reactive phosphorus group, and R2 is an H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or a C. 1-30Hydrocarbon group or solid support. Preferably, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group. More preferably, R1 is a reactive phosphorus group, and R2 is a C24-hydroxyl protecting group. 1-30 Hydrocarbon group. More preferably, R1 is a reactive phosphorus group, and R2 is a hydroxyl protecting group (such as DMTr). More preferably, both R1 and R2 are reactive phosphorus groups, and the reactive phosphorus groups may be the same or different. More preferably, R1 is a solid support, and R2 is a hydroxyl protecting group (such as DMTr).

[0241] Optional, where B is -L B -(C 1-30 The skeletal structure of a compound of formula (Id) with a hydrocarbon group is added to the 5' end and / or 3' end (preferably the 5' end or 3' end of the sense strand) of the sense and / or antisense strands of the dsRNA activator.

[0242] In some embodiments, compounds of formula (Ie) or salts thereof (e.g., pharmaceutically acceptable salts), solvates (e.g., hydrates), or isotopically labeled compounds (e.g., deuterated compounds) are provided:

[0243] in:

[0244] B is a base (including modified or unmodified bases);

[0245] R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 hydrocarbon group or C 1-30 Hydrocarbon group; optionally, R1 and R2 are not both C 1-30 hydrocarbon group;

[0246] R3 is C 1-30 hydrocarbon group;

[0247] R B For H or C 1-6 Alkyl groups, preferably H;

[0248] Wherein C 1-30 The hydrocarbon group may be fully saturated or partially unsaturated. Optionally, the variables are further defined as herein. Optionally, the C 1-30 The hydrocarbon group is C 12-30 hydrocarbon group, C 14-24 hydrocarbon group or C 16-22 The hydrocarbon group is fully saturated or partially unsaturated, and / or the hydrocarbon group is straight-chain or branched. Optionally, the hydrocarbon group is C24. 1-30 Alkyl, such as C 12-30 Alkyl, C 14-24 Alkyl or C 16-22Alkyl group, optionally straight-chain or branched. Optionally, the hydrocarbon group is C10. 2-30 alkenyl groups, such as C 12-30 alkenyl, C 14-24 alkenyl or C 16-22 Alkenyl group, optionally the alkenyl group is straight-chain or branched. Optionally, the alkynyl group is C10. 2-30 Alkyne groups, such as C 12-30 alkynyl group, C 14-24 alkynyl or C 16-22 The alkynyl group, optionally, is straight-chain or branched.

[0249] Optionally, the reactive phosphorus group is derived from a phosphate ester group, a thiophosphate ester group, an O,S-thiophosphate ester group, or a phosphoramidite group.

[0250] In some embodiments, R1 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 A hydrocarbon group or solid support, and R2 is a reactive phosphorus group. In some embodiments, R1 is a reactive phosphorus group, and R2 is an H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or a C. 1-30 Hydrocarbon group or solid support. Preferably, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group. More preferably, R1 is a reactive phosphorus group, and R2 is a C24-hydroxyl protecting group. 1-30 Hydrocarbon group. More preferably, R1 is a reactive phosphorus group, and R2 is a hydroxyl protecting group (such as DMTr). More preferably, both R1 and R2 are reactive phosphorus groups, and the reactive phosphorus groups may be the same or different. More preferably, R1 is a solid support, and R2 is a hydroxyl protecting group (such as DMTr).

[0251] Optionally, the backbone structure of the compound of formula (Ie), where B is a base (including modified or unmodified bases), is used to replace the original nucleotide at an internal position of the sense and / or antisense strands.

[0252] In some embodiments, the compound of formula (I) has a structure of formula (If) or (If'), preferably having the structure of formula (If).

[0253] B, X, and R3 are defined as above, for example, as defined in equation (I).

[0254] In some embodiments, in formula (If) or (If'), B is H or a base (including modified or unmodified bases), optionally said modified or unmodified bases are further defined as herein.

[0255] In some embodiments, in formula (If) or (If'), X is selected from -O-, *-O-NR4-, *-NR4-O-, and *-O-NR4-CO-, preferably selected from -O-, *-ON(CH3)-, *-N(CH3)-O-, *-O-NH-CO-, and *-ON(CH3)-CO-, wherein the valence bond indicated by * is connected to the pentose ring, and R3 and B are as defined herein.

[0256] In some implementations, R3 is C in equation (If) or (If'). 1-30 Hydrocarbon group, optionally the C 1-30 The hydrocarbon group is further defined as in this article.

[0257] In some implementations, in equation (If) or (If'), B is H, X is -O-, and R3 is C. 1-30 Hydrocarbon group.

[0258] In some embodiments, in formula (If) or (If'), B is a base (including modified or unmodified bases); X is selected from -O-, *-O-NR4-, *-NR4-O-, and *-O-NR4-CO-, preferably selected from -O-, *-ON(CH3)-, *-N(CH3)-O-, *-O-NH-CO-, and *-ON(CH3)-CO-, wherein the * indicates a valence bond connected to a pentose ring; and R3 is C 1-30 Hydrocarbon group.

[0259] In some embodiments, the compound of formula (I) has the structures of formulas (If-1), (If-2), (If-3), and (Ife-4).

[0260] Where B is H or as defined above, preferably H or a base (including modified or unmodified bases).

[0261] In some embodiments, the compound of formula (I) has the structure of the following formula:

[0262] X and R3 are defined as above, for example, as defined in equation (If).

[0263] In some embodiments, the compound of formula (I) has the structure of formula (Ig) or (Ig'), preferably the structure of formula (Ig):

[0264] Wherein, B is as defined herein. Preferably, B is -L B -(C 1-30 Hydrocarbon group). Optionally, the -L B -(C 1-30(Hydrocarbon group) is further defined as in this article.

[0265] In some embodiments, the compound of formula (I) is selected from:

[0266] II. dsRNA activator

[0267] In some aspects, the present invention provides RNAi activators that inhibit target genes, such as dsRNA activators, wherein the RNAi activator comprises the backbone structure of the compounds of the present invention as defined in Part I at one or more positions on at least one strand. In some embodiments, the dsRNA activator is siRNA. In some embodiments, the siRNA comprises a double-stranded ribonucleic acid (dsRNA) molecule for inhibiting the expression of target genes in cells.

[0268] In some embodiments, the target gene is present (e.g., predominantly present) in specific tissues or cells, such as muscle tissue (e.g., skeletal muscle tissue or cardiac muscle tissue) or cells, or adipose tissue or cells. In some embodiments, the target gene is, for example, the SOD1 or ACVR1C gene.

[0269] Intrinsic RNAi (RNA interference) mechanisms in organisms typically involve a series of processes, including: Dicer processing long dsRNA into short 19-21 base pairs (bp) siRNA; the siRNA binding to the Ago protein to form an RNA-induced silencing complex (RISC); the Ago protein cleaving the sense strand of the siRNA and releasing it; subsequently, the mature RISC bound to the antisense strand cleaves the mRNA that is anticomplementary to the antisense strand through a sequence complementation mechanism. Based on this RNA interference mechanism, various artificial RNAi molecules with different structures have been developed. These structures can enter the RNAi pathway at different stages to achieve sequence-specific cleavage of target gene transcripts. See, for example, Molecules 2019, 24, 2211; doi:10.3390 / molecules24122211 (in its entirety incorporated herein by reference). Artificial RNAi molecules with such structures include, for example, siRNA molecules having a double-stranded region (and optionally one or two overhangs), long-chain siRNA molecules that can serve as substrates for the Dicer enzyme, short hairpin RNAs (shRNAs) that can be processed by Dicer to produce siRNA structures, and long single-stranded siRNA molecules containing only an antisense strand. It is understood that these molecular forms all fall within the scope of the RNAi activators of the present invention. Furthermore, siRNA or dsRNA may contain modified nucleotides, and may also contain ligands that deliver siRNA or dsRNA to targeted tissues or cells in vivo. These dsRNAs or siRNAs with modified nucleotides, as well as dsRNAs and siRNAs with modified nucleotides and ligands, are also dsRNA activators of the present invention.

[0270] In some embodiments, the dsRNA activator of the present invention, such as siRNA (including siRNA having modified nucleotides and siRNA having modified nucleotides and the backbone structure of the compounds of the present invention), inhibits the expression of target genes (e.g., the SOD1 gene or the ACVR1C gene) at least 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99%, for example in specific tissues or cells, such as muscle tissue (e.g., skeletal muscle tissue or cardiac tissue) or cells, or adipose tissue or cells, as determined by, for example, PCR or by protein-based methods (e.g., by immunofluorescence analysis, using, for example, Western blotting or flow cytometry techniques). In some embodiments, the inhibition rate of expression is determined in suitable biological cell lines (e.g., muscle cells or adipocytes) using the qPCR method provided herein, for example, with a concentration of dsRNA such as siRNA at approximately 10 nM. In some embodiments, the inhibition rate of expression is determined in suitable biological cell lines using, for example, serially diluted concentrations of dsRNA such as siRNA, using the qPCR method provided herein.

[0271] In some embodiments, the dsRNA activators of the present invention, such as siRNA (including siRNA with modified nucleotides and siRNA with modified nucleotides and the backbone structure of the compounds of the present invention), are capable of effectively inhibiting the expression of target genes in vivo (e.g., in specific tissues or cells), such as in muscle tissue (e.g., skeletal muscle tissue or cardiac tissue) or cells, or in adipose tissue or cells.

[0272] In some embodiments, the dsRNA activator of the present invention, such as siRNA (including siRNA having modified nucleotides and siRNA having modified nucleotides and the backbone structure of the compounds of the present invention), inhibits the expression of target genes (e.g., the SOD1 gene or the ACVR1C gene) in vivo (e.g., in specific tissues or cells) by at least about 30%, 35%, about 40%, 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%, for example by detection of homogenates of mouse adipose tissue (e.g., subcutaneous fat and / or gonadal fat, or muscle tissue such as skeletal muscle (e.g., quadriceps femoris) or myocardial tissue or cells) extracted after a single intravenous administration to mice, as described in, for example, as in Examples 4, 5, 8, or 9.

[0273] In some embodiments, the dsRNA activator of the present invention comprises an antisense strand containing a complementary region that is complementary (substantially complementary or fully complementary) to at least a portion (e.g., the target sequence) of the mRNA formed during the expression of a target gene (e.g., the SOD1 gene or the ACVR1C gene). In some embodiments, the length of the complementary region is about 15 to 30 nucleotides, such as 16 to 30 nucleotides, 17 to 30 nucleotides, or 18 to 30 nucleotides (e.g., lengths of about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, or 15 nucleotides). In some embodiments, the length of the complementary region is between 18 and 23 nucleotides. In some embodiments, the length of the complementary region is 19 to 23 nucleotides. In some embodiments, the length of the complementary region is 18 to 21 nucleotides. In some embodiments, the length of the complementary region is 18, 19, 20, 21, 22, or 23 nucleotides. In some embodiments, the length of the complementary region is 15, 16, 17, 18, 19, 20, 21, 22, or 23 nucleotides. In some embodiments, the antisense strand is complementary to the mRNA target sequence starting from the first or second nucleotide from the 5' end. In some embodiments, the complementary region of the antisense strand comprises the first or second nucleotide from the 5' end to the third, second, or first nucleotide from the 3' end. In some embodiments, the complementary region of the antisense strand comprises all antisense strand nucleotides starting from the second nucleotide from the 5' end. In some embodiments, the complementary region of the antisense strand comprises at least the following nucleotides, starting from the 5' end: nucleotides 1-16, 1-17, 1-18, 1-19, 1-20, 1-21, 1-22, 1-23, 2-16, 2-17, 2-18, 2-19, 2-20, 2-21, 2-22, or 2-23. In some embodiments, the complementary region of the antisense strand comprises at least the following nucleotides, starting from the 5' end: nucleotides 2-19. In some implementations, the complementary region of the antisense strand comprises, or is composed of, consecutive nucleotides at positions 1-19, 1-20, 1-21, 1-22, 1-23, 2-19, 2-20, 2-21, 2-22, or 2-23, starting from the 5' end of the antisense strand.

[0274] In some implementations, the dsRNA comprises two complementary RNA strands that form a double-stranded structure (double-stranded region or double-stranded region) under conditions that will cause the dsRNA to hybridize, namely the antisense strand and the sense strand.

[0275] In some embodiments, the antisense strand of the dsRNA includes a complementary region (antisense complement) that is substantially or completely complementary to the target sequence. Therefore, the antisense complement of the dsRNA can be substantially or completely complementary to the target sequence. The target sequence can be derived from the sequence of mRNA formed during the expression of the target gene (e.g., the SOD1 gene or the ACVR1C gene). In some embodiments, the antisense complement is substantially complementary to the target sequence, for example, it is mismatched with the target sequence at 1, 2, 3, 4, or 5 nucleotides (preferably 1 or 2 nucleotides at the 5' end and / or the 3' end, e.g., the first nucleotide at the 5' end of the antisense strand). In some embodiments, the antisense complement is completely complementary to the target sequence.

[0276] In some embodiments, the antisense strand of the dsRNA, starting from the second nucleotide from the 5' end, is completely complementary to the corresponding portion of the target sequence. In some embodiments, the antisense strand of the dsRNA, from the second nucleotide from the 5' end to the first, second, or third nucleotide from the 3' end, is completely complementary to the corresponding portion of the target sequence. In some embodiments, the entire length of the antisense strand of the dsRNA, starting from the second nucleotide from the 5' end, is completely complementary to the corresponding portion of the target sequence. In some embodiments, nucleotides 2-16, 2-17, 2-18, 2-19, 2-20, 2-21, 2-22, or 2-23 of the antisense strand of the dsRNA, starting from the 5' end, are completely complementary to the corresponding portion of the target sequence. In some embodiments, consecutive nucleotides from positions 2-19, 2-20, 2-21, 2-22, or 2-23 of the antisense strand of the dsRNA, starting from the 5' end, are completely complementary to the corresponding portion of the target sequence. In some embodiments, the antisense strand of the dsRNA has the same number of nucleotides as the target sequence and is completely complementary to the corresponding portion of the target sequence in all nucleotide sequences except for the first nucleotide at the 5' end, wherein the first nucleotide of the antisense strand is U or A.

[0277] In some embodiments, the antisense strand of the dsRNA is completely complementary to the corresponding portion of the target sequence, starting from the first nucleotide at the 5' end. In some embodiments, the antisense strand of the dsRNA is completely complementary to the corresponding portion of the target sequence from the first nucleotide at the 5' end to the first, second, or third nucleotide at the 3' end. In some embodiments, the entire length of the antisense strand of the dsRNA, starting from the first nucleotide at the 5' end, is completely complementary to the corresponding portion of the target sequence. In some embodiments, nucleotides 1-16, 1-17, 1-18, 1-19, 1-20, 1-21, 1-22, or 1-23 of the antisense strand of the dsRNA, starting from the 5' end, are completely complementary to the corresponding portion of the target sequence. In some embodiments, the antisense strand of the dsRNA contains consecutive nucleotides at positions 1-19, 1-20, 1-21, 1-22, or 1-23, starting from the 5' end, that are completely complementary to the corresponding portion of the target sequence. In some embodiments, the full length of the antisense strand is completely complementary to the target sequence.

[0278] When referring to "the corresponding portion of the target sequence" in this document, "the corresponding portion of the target sequence" means a consecutive nucleotide sequence in the target sequence that is completely complementary to the antisense strand. For example, when the target sequence is 21 nucleotides and its consecutive nucleotides from position 1 to 20, counting from the 5' end, are completely complementary to the consecutive nucleotides from position 2 to 21, counting from the 5' end, the "corresponding portion of the target sequence" refers to the consecutive nucleotides from position 1 to 20, counting from the 5' end, of the target sequence.

[0279] In some implementations, the target sequence comprises, or is composed of, a continuous sequence of the mRNA of a target gene (e.g., the SOD1 gene or the ACVR1C gene).

[0280] In some implementations, the antisense strand contains 1, 2, 3, 4, or 5 non-complementary sites (mismatches) with the target sequence, for example, mismatches at 1-3 nucleotides. In some implementations, the antisense strand is completely complementary to the target sequence.

[0281] In some embodiments, the antisense strand of the dsRNA is completely complementary to the target sequence of the target gene (e.g., the SOD1 gene or the ACVR1C gene) except for one, two, or three nucleotides. In some embodiments, the antisense strand of the dsRNA is completely complementary to the target sequence of the target gene (e.g., the SOD1 gene or the ACVR1C gene) in the region excluding the first and / or second nucleotides at the 5' end, and to the target sequence in the region excluding the first and / or second nucleotides at the 3' end. In some embodiments, the antisense strand of the dsRNA is completely complementary to the target sequence of the target gene (e.g., the SOD1 gene or the ACVR1C gene) in the region excluding the first nucleotide at the 5' end, and to the target sequence in the region excluding the first nucleotide at the 3' end.

[0282] In some embodiments, the first nucleotide at the 5' end of the antisense strand of the dsRNA is U or A, for example, U, to facilitate recognition by the Ago2 protein to form the RICS complex.

[0283] In some embodiments, the sense strand of the dsRNA of the present invention includes a region complementary to the antisense strand, such that the two strands hybridize and form a double-stranded structure (double-stranded region) when combined under appropriate conditions.

[0284] In some embodiments, the length of the sense strand and the antisense strand is independently 15-30 nucleotides, such as 17-27, 19-25, 18-24, 18-23, 19-22, or 19-21 nucleotides. In some embodiments, the length of the antisense strand or sense strand is independently no more than 27, 26, 25, 24, 23, 22, 21, 20, or 19 nucleotides. In some embodiments, the length of the antisense strand or sense strand is independently not less than 15, 16, 17, 18, 19, 20, or 21 nucleotides. In some embodiments, the length of the sense strand is 18-21 nucleotides (e.g., 18, 19, 20, or 21 nucleotides), and the length of the antisense strand is 19-23 nucleotides (e.g., 19, 20, 21, 22, or 23 nucleotides). In some embodiments, the length of the sense strand is 18, 19, 20, or 21 nucleotides, and the length of the antisense strand is 19-23 nucleotides. In some embodiments, the length of the sense strand is 19 nucleotides, and the length of the antisense strand is 21 nucleotides. In some embodiments, the length of the sense strand is 21 nucleotides, and the length of the antisense strand is 21 nucleotides. In some embodiments, the length of the sense strand is 21 nucleotides, and the length of the antisense strand is 23 nucleotides.

[0285] In some embodiments, the antisense and sense strands hybridize to form a double-stranded region. In some embodiments, the length of the double-stranded region is 15 to 30 nucleotide pairs. In some embodiments, the length of the double-stranded region is 15 to 25 nucleotide pairs or 16 to 25 nucleotide pairs. In some embodiments, the length of the double-stranded region is 16 to 24 nucleotide pairs or 17 to 24 nucleotide pairs. In some embodiments, the length of the double-stranded region is 17 to 23 nucleotide pairs or 18 to 23 nucleotide pairs. In some embodiments, the length of the double-stranded region is 16 to 22 nucleotide pairs, 17 to 22 nucleotide pairs, 18 to 22 nucleotide pairs, or 19 to 22 nucleotide pairs. In some embodiments, the length of the double-stranded region is 16 to 21 nucleotide pairs, for example, 16, 17, 18, 19, 20, or 21 nucleotide pairs. In some embodiments, the length of the double-stranded region is 18 to 21 nucleotide pairs. In some implementations, the length of the double-stranded region is 18, 19, 20, or 21 nucleotide pairs.

[0286] In some embodiments, the double-stranded region formed by the sense and antisense strands is completely complementary. In other embodiments, the double-stranded region formed by the sense and antisense strands is substantially complementary, and may contain one, two, three, four, or five non-complementary sites (mismatches), for example, located at the 5' and / or 3' ends of the sense and / or antisense strands or located within the sense and / or antisense strands. In some embodiments, non-complementary sites (base mismatches) are included between the antisense and sense strands due to the introduction of the compound of the present invention into the antisense or sense strand.

[0287] In some embodiments, the length of the fully complementary double-stranded region is at least 13, 14, 15, 16, 17, 18, or 19 nucleotides. In some embodiments, the length of the fully complementary double-stranded region is between 15 and 25 nucleotide pairs, 16 and 25 nucleotide pairs, 16 and 24 nucleotide pairs, 17 and 24 nucleotide pairs, 17 and 23 nucleotide pairs, 18 and 23 nucleotide pairs, or 19 and 22 nucleotides. In some embodiments, the length of the fully complementary double-stranded region is 16, 17, 18, 19, 20, or 21 nucleotides.

[0288] In some embodiments, the number of completely complementary nucleotide pairs in the double-stranded region is at least 15, 16, 17, 18, 19, 20, or 21 nucleotide pairs. In some embodiments, the number of completely complementary nucleotide pairs in the double-stranded region is between 15 and 25, 16 and 25, 16 and 24, 17 and 24, 17 and 23, 18 and 23, or 19 and 22 nucleotide pairs. In some embodiments, the number of completely complementary nucleotide pairs in the double-stranded region is 16, 17, 18, 19, 20, or 21 nucleotide pairs.

[0289] The dsRNA described herein may further comprise one or more single-stranded nucleotide overhangs, for example, 1 to 4, 2 to 4, 1 to 3, 2 to 3, 1, 2, 3, or 4 nucleotides. In some embodiments, dsRNA having at least one nucleotide overhang has better repressive properties relative to its blunt-ended counterpart. The nucleotide overhang may include or consist of nucleotide / nucleoside analogs, including deoxynucleotides / nucleosides. The overhang may be on the sense strand, antisense strand, or any combination thereof. Furthermore, the overhanging nucleotide may be present at the 5' end, 3' end, or both ends of the antisense strand or sense strand of the dsRNA.

[0290] In some embodiments, one or both of the sense strand and the antisense strand include a 3' overhang and / or a 5' overhang having at least 1, 2, or 3 nucleotides; for example, one or both of the sense strand and the antisense strand include a 3' overhang and / or a 5' overhang having at least 1 nucleotide. In some embodiments, at least one strand includes a 3' overhang or a 5' overhang having at least 1 nucleotide. In some embodiments, at least one strand includes a 3' overhang or a 5' overhang having at least 2 nucleotides. In some embodiments, at least one strand includes a 3' overhang or a 5' overhang having at least 3 nucleotides.

[0291] In some preferred embodiments, the antisense strand has a 3' overhang and / or a 5' overhang of at least one nucleotide, for example, the antisense strand comprises a 3' overhang and / or a 5' overhang of one nucleotide. In some preferred embodiments, the antisense strand has a 3' overhang and / or a 5' overhang of at least two nucleotides, for example, the antisense strand comprises a 3' overhang and / or a 5' overhang of two nucleotides. In some preferred embodiments, the antisense strand has a 3' overhang and / or a 5' overhang of at least three nucleotides, for example, the antisense strand comprises a 3' overhang and / or a 5' overhang of three nucleotides. In a preferred embodiment, the antisense strand has a 3' overhang of one, two, or three nucleotides at the 3' end, for example, a 3' overhang of two nucleotides.

[0292] In some embodiments, the sense strand includes a 5' overhang having at least 1, 2, or 3 nucleotides, and / or the antisense strand includes a 3' overhang having at least 1, 2, or 3 nucleotides.

[0293] In some embodiments, the antisense strand of the dsRNA has a 3' overhang, for example, a 2-nucleotide 3' overhang, and a blunt end at the 5' end. In some embodiments, both the antisense and sense strands of the dsRNA are blunt at both ends.

[0294] In some embodiments, the present invention relates to a double-stranded RNA (dsRNA) activator for inhibiting the expression of a target gene (e.g., the SOD1 gene or the ACVR1C gene), wherein the dsRNA activator comprises a sense strand and an antisense strand forming a double-stranded region, wherein the antisense strand is completely complementary to the corresponding portion of the target sequence of the target gene (e.g., the SOD1 gene or the ACVR1C gene) at least from the 5' end at positions 2-19 (e.g., positions 2-20, 2-21, 2-22, 2-23, or the full length), for example, wherein the antisense strand is completely complementary to the target sequence of the target gene (e.g., the SOD1 gene or the ACVR1C gene) in a region other than the first nucleotide at the 5' end, wherein the first nucleotide at the 5' end of the antisense strand is A or U, for example, U.

[0295] In some embodiments, the dsRNA activator of the present invention is conjugated with one or more long-chain hydrocarbon chains at one or more positions on the sense and / or antisense strands, for example by replacing the nucleotides at said positions with a compound backbone structure of the present invention bearing said long-chain hydrocarbon chains, or by adding a compound backbone structure bearing said long-chain hydrocarbon chains to the 5' or 3' end of the sense and / or antisense strands. In some embodiments, the conjugated long-chain hydrocarbon chains make the resulting dsRNA activator suitable for delivery to specific tissues or cells, such as muscle tissue (e.g., skeletal muscle tissue or cardiac muscle tissue) or cells, and adipose tissue or cells.

[0296] In some embodiments, the hydrocarbon chain may be saturated or unsaturated and may be straight-chain or branched, and may optionally be substituted with groups selected from: hydroxyl, amino, alkylamino, sulfonate, phosphate, mercapto, azide, and alkynyl. In some embodiments, the hydrocarbon chain conjugated to one or more positions on the at least one chain is C 1-30 Hydrocarbon groups, such as those defined herein, such as C 14-24 Hydrocarbon group, preferably C 16 -C 22 Hydrocarbon groups, such as C 16 C 17 C 18 C 19 C 20 C 21 Or C 22 Hydrocarbon groups, such as hexadecyl, hexadecylene, hexadecadienyl, heptadecanyl, heptadecanyl, heptadecanyl, octadecyl, octadecylene, octadecadienyl, octadectrienyl, nonadecanyl, nonadecanylene, nonadecanadienyl, nonadecantrienyl, eicosyl, eicosylene, eicosyladienyl, eicosyltrienyl, dodecyl, dodecylene, dodecylene, dodecylene, 6-octyltetradecyl, 10-hexylhexadecyl, all cis 7,10,13,16,19-docosapentaenyl, all cis 4,7,10,13,16,19-docosahexaenyl, all cis 13,16-docosadienyl, all-cis-7,10,13,16-docosatetraenyl, all-cis-4,7,10,13,16-docosapentenyl or cis-13-docosaenyl, particularly 1-docoalkyl, 6-octyltetradecane-1-yl, 10-hexylhexadecane-1-yl, cis-docosa-13-en-1-yl, docosane-9-yl, docosane-2-yl, docosane-10-yl, docosane-11-yl or cis-4,7,10,13,16,19-docosahexaen-1-yl, wherein the hydrocarbon group may optionally be substituted with a group selected from the following: hydroxyl, amino, cyano, nitro, halogen, C 1- 6-alkoxy, C 3-8 Cyclic hydrocarbon group, C 3-8 Cycloalkyloxy group, 3-8 membered heterocyclic group, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, mercapto, azide, alkynyl, aryl, or heteroaryl, or optionally, a C10 group is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6Cycloalkyl. In some embodiments, the hydrocarbon chain conjugated to one or more positions on the at least one chain is 1-hexadecyl, 1-eicosyl, 1-monoalkyl, or 1-docoalkyl.

[0297] In some embodiments, the dsRNA activator of the present invention comprises, at one or more positions on the sense and / or antisense strands, the skeletal structure of a compound of formula (I) or any subform thereof such as formulas (Ia) to (If) of the present invention, or the skeletal structure of a specific compound synthesized in Example 1, wherein the skeletal structure of the compound of formula (I) is as follows:

[0298] The variables are defined as described in this paper (e.g., Part I).

[0299] The skeletal structure of the compound is introduced by adding the skeletal structure to the 5' and / or 3' ends of the sense and / or antisense strands, or by replacing one or more internal positions of the sense and / or antisense strands with the skeletal structure, wherein when the corresponding structure is used to replace nucleotides at one or more internal positions of the sense and / or antisense strands, And ------ are respectively linked to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide; preferably, Linked to the 3' carbon or corresponding position of the previous nucleotide, ------ linked to the 5' carbon or corresponding position of the next nucleotide; or when the corresponding structure is added to the 5' or 3' end of the sense and / or antisense strands. One of the two symbols is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other symbol represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is added to the 5' end of the sense strand and / or antisense strand, Linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ linked to the 5' carbon of the 5' terminal nucleotide or the corresponding position; or when the corresponding structure is added to the 3' end of the sense strand and / or antisense strand, ------ linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position.

[0300] In some embodiments, the compound of formula (I) has formula (Ia), and its skeletal structure is shown in the following formula:

[0301] In some embodiments, the compound of formula (I) has a (Ib) structure, and its skeletal structure is shown in the following formula:

[0302] In some embodiments, the compound of formula (I) has the structure of formula (Ic), and its skeletal structure is shown in the following formula:

[0303] In some embodiments, the compound of formula (I) has the structure of formula (Id), and its skeletal structure is shown in the following formula:

[0304] In some embodiments, the compound of formula (I) has the structure of formula (Ie), and its skeletal structure is shown in the following formula:

[0305] In some embodiments, the sense and / or antisense strands of the dsRNA activator of the present invention comprise, at one or more positions, the skeletal structure of a compound of formula (I) or any subform thereof such as formulas (Ia) to (Ig) of the present invention, or the skeletal structure of a specific compound synthesized in Example 1, wherein the skeletal structure of the compound of formula (I) is shown in formula (I”) or any subform thereof.

[0306] The variables are as defined herein, and the backbone structure of the compound is introduced by adding the backbone structure to the 5' and / or 3' ends of the sense and / or antisense strands, or by replacing one or more internal positions of the sense and / or antisense strands with the backbone structure, wherein when the corresponding structure is used to replace nucleotides at one or more internal positions of the sense and / or antisense strands, And ------ are respectively linked to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide; preferably, Linked to the 3' carbon or corresponding position of the previous nucleotide, ------ linked to the 5' carbon or corresponding position of the next nucleotide; or when the corresponding structure is added to the 5' or 3' end of the sense and / or antisense strands. One of the two symbols is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other symbol represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is added to the 5' end of the sense strand and / or antisense strand, Linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ linked to the 5' carbon of the 5' terminal nucleotide or the corresponding position; or when the corresponding structure is added to the 3' end of the sense strand and / or antisense strand, ------ linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position.

[0307] In some embodiments, nucleotides added to or located at the 5' or 3' end of the sense strand and / or antisense strand, or at internal positions of the sense strand, are replaced with one or more structures independently selected from the following:

[0308] Wherein, Y is oxygen or sulfur, and wherein, when the corresponding structure is used to replace a nucleotide at one or more internal positions of the sense and / or antisense strands, And ------ are respectively linked to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide; preferably, Linked to the 3' carbon or corresponding position of the previous nucleotide, ------ linked to the 5' carbon or corresponding position of the next nucleotide; or

[0309] When the corresponding structure is added to the 5' or 3' end of the justice chain and / or antisense chain, One of the two symbols is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other symbol represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is added to the 5' end of the sense strand and / or antisense strand, Linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ linked to the 5' carbon of the 5' terminal nucleotide or the corresponding position; or when the corresponding structure is added to the 3' end of the sense strand and / or antisense strand, ------ linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position.

[0310] In some embodiments, the skeletal structures corresponding to ND001, ND002, ND003, ND009, ND010, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, and ND060 are shown below, wherein the compounds have the same ND number as their skeletal structures:

[0311] Wherein, Y is oxygen or sulfur, and wherein, when the corresponding structure is used to replace a nucleotide at one or more internal positions of the sense and / or antisense strands, And ------ are respectively linked to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide; preferably, Linked to the 3' carbon or corresponding position of the previous nucleotide, ------ linked to the 5' carbon or corresponding position of the next nucleotide; or when the corresponding structure is added to the 5' or 3' end of the sense and / or antisense strands. One of the two symbols is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other symbol represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is added to the 5' end of the sense strand and / or antisense strand, Linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ linked to the 5' carbon of the 5' terminal nucleotide or the corresponding position; or when the corresponding structure is added to the 3' end of the sense strand and / or antisense strand, ------ linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position.

[0312] In some embodiments, nucleotides added to or located at one or more internal positions of the positive and / or negative strands at the 5' or 3' end are replaced with one or more structures independently selected from the following:

[0313] Wherein, when the corresponding structure is used to replace nucleotides at one or more internal positions of the sense strand and / or antisense strand, And ------ are respectively linked to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide; preferably, Linked to the 3' carbon or corresponding position of the previous nucleotide, ------ linked to the 5' carbon or corresponding position of the next nucleotide; or when the corresponding structure is added to the 5' or 3' end of the sense and / or antisense strands. One of the two symbols is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other symbol represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is added to the 5' end of the sense strand and / or antisense strand, Linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ linked to the 5' carbon of the 5' terminal nucleotide or the corresponding position; or when the corresponding structure is added to the 3' end of the sense strand and / or antisense strand, ------ linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position.

[0314] In some embodiments, the skeletal structures corresponding to ND001, ND002, ND003, ND009, ND010, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, and ND060 are shown below, wherein the compounds have the same ND number as their skeletal structures:

[0315] Wherein, when the corresponding structure is used to replace nucleotides at one or more internal positions of the sense strand and / or antisense strand, And ------ are respectively linked to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide; preferably, Linked to the 3' carbon or corresponding position of the previous nucleotide, ------ linked to the 5' carbon or corresponding position of the next nucleotide; or

[0316] When the corresponding structure is added to the 5' or 3' end of the justice chain and / or antisense chain, One of the two symbols is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other symbol represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is added to the 5' end of the sense strand and / or antisense strand, Linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ linked to the 5' carbon of the 5' terminal nucleotide or the corresponding position; or when the corresponding structure is added to the 3' end of the sense strand and / or antisense strand, ------ linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ It is attached to the 3' carbon of the 3' terminal nucleotide or the corresponding position. It should be understood that the structure of ND010' depends on whether it is added to the 5' end or the 3' end. When it is added to the 5' end of the positive and / or negative strands, the structure of ND010' is shown as ND010'-2; when it is added to the 3' end of the positive and / or negative strands, the structure of ND010' is shown as ND010'-1.

[0317] In some embodiments, the dsRNA activator of the present invention is, at one or more positions on the positive strand, a compound of the present invention, such as a compound of formula (I) or any subform thereof of the present invention, such as formulas (Ia) to (If), or the skeletal structure of a specific compound synthesized in Example 1, wherein the skeletal structure of the compound is introduced into the positive strand by adding the skeletal structure to the 5' end and / or 3' end of the positive strand, or by replacing the skeletal structure at one or more internal positions on the positive strand.

[0318] In some embodiments, the backbone structure of the compound of the present invention is added to the 5' end and / or 3' end (preferably the 5' end or 3' end of the sense strand) of the dsRNA activator, for example, by linking it to the 5' end or 3' end nucleotide of the sense strand via a phosphate ester bond or a thiophosphate ester bond. In some specific embodiments, when the backbone structure of the compound of the present invention is added to the 5' end and / or 3' end (preferably the 5' end or 3' end of the sense strand) of the sense strand and / or antisense strand of the dsRNA activator, the B in the compound is selected from H, a base (e.g., a modified or unmodified base), or -L. B -(C 1-30 (Hydrocarbon group). In some more specific embodiments, when the backbone structure of the compound of the present invention is added to the 5' end and / or 3' end (preferably the 5' end or 3' end of the sense strand or antisense strand) of the dsRNA activator, the B in the compound is selected from H or -L. B -(C 1-30(Hydrocarbon group). In some specific embodiments, the skeletal structure of the compound that can be added to the 5' end and / or 3' end (preferably the 5' end or 3' end of the sense strand) of the sense strand and / or antisense strand of the dsRNA activator is selected from the skeletal structures of the following compounds: ND001, ND002, ND003, ND009, ND010, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, ND060. In some more specific embodiments, the skeletal structure of the compound that can be added to the 5' end and / or 3' end (preferably the 5' end or 3' end of the sense strand) of the sense strand and / or antisense strand of the dsRNA activator is preferably selected from the skeletal structures of the following compounds: ND009, ND010, ND030, ND031, ND032, ND033, ND036, ND045, ND046, ND047 and ND048.

[0319] In some more specific embodiments, the backbone structure of ND001, ND002, ND003, ND009, ND010, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, or ND060 may be added to the 5' and / or 3' ends (preferably the 5' end or the 3' end of the sense strand) of the dsRNA activator. In some embodiments, the skeleton structure is as shown in ND001', ND002', ND003', ND009', ND010', ND011', ND028', ND029', ND030', ND031', ND032', ND033', ND0036', ND042', ND0045', ND0046', ND0047', ND0048', ND052', ND053', ND054', ND055', ND056', ND057', ND058', or ND060'.

[0320] In some more specific embodiments, the backbone structure of ND010, ND009, ND036, ND031, ND048, or ND046 may be added to the 5' and / or 3' ends (preferably the 5' or 3' end of the sense strand) of the sense and / or antisense strands of the dsRNA activator. In some embodiments, the backbone structure is as shown in ND010', ND009', ND036', ND031', ND048', or ND046'.

[0321] In some embodiments, the backbone structure of the compound of the present invention replaces the original nucleotide at one or more internal positions on the sense and / or antisense strands. In some embodiments, the internal positions are selected from any one or more positions other than the three terminal positions at each end of at least one strand. In some embodiments, the internal positions are not located within the cleavage site region of the sense strand. In some embodiments, the internal positions are not positions 9-12 or 11-13 counting from the 5' end of the sense strand, for example, not or not included in any of these positions.

[0322] In some embodiments, the internal location does not include the cleavage site region of the antisense strand. In some embodiments, the internal location is not a location within the cleavage site region of the antisense strand. In some embodiments, the internal location is not the 12th-14th position counting from the 5' end of the antisense strand, for example, it is not or does not include any of those positions.

[0323] In some embodiments, the compound of formula (I) of the present invention is located at one or more of the following positions on the justice chain: positions 1-8 and 13-18 on the justice chain, for example, positions 1, 5, 6, 7, 15 on the justice chain, counted from the 5' end of the justice chain.

[0324] In some embodiments, the compound of formula (I) of the invention is located at one or more of the following positions on the antisense chain: positions 6-10 and 15-18 on the antisense chain, for example, positions 15 and 17 on the antisense chain, counted from the 5' end of the antisense chain.

[0325] In some implementations, the internal position is the 6th position counting from the 5' end of the justice chain.

[0326] In some embodiments, when the backbone structure of the compound of the present invention is used to replace nucleotides at one or more internal positions of the sense and / or antisense strands, the B in the compound is preferably H, a base (e.g., a modified or unmodified base), or -L. B -(C 1-30The base (H) is preferably a base complementary to the base of the nucleotide at the corresponding position on the other chain (e.g., a modified or unmodified base). For example, if the skeletal structure of the compound of the present invention is at position 6 from the 5' end on the sense chain, then the B in the compound is preferably an unmodified base complementary to the base of the nucleotide at the corresponding position on the antisense chain, or H.

[0327] In some specific embodiments, the backbone structure of the compound that can be used to replace the internal position of the pronucleotide in the sense strand and / or antisense strand is selected from the backbone structures of the following compounds: ND001, ND002, ND003, ND009, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, ND060; preferably selected from the backbone structures of the following compounds: ND001, ND002, ND009, ND011, ND032 or ND033.

[0328] In some more specific implementations, the backbone structure of ND001, ND002, ND003, ND009, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, or ND060 can be used to replace the original nucleotides at internal positions of the sense and / or antisense strands. In some embodiments, the skeleton structure is as shown in ND001', ND002', ND003', ND009', ND011', ND028', ND029', ND030', ND031', ND032', ND033', ND0036', ND042', ND0045', ND0046', ND0047', ND0048', ND052', ND053', ND054', ND055', ND056', ND057', ND058', or ND060'.

[0329] In some more specific embodiments, the backbone structure of ND001, ND002, ND009, ND011, ND032, or ND033 can be used to replace the original nucleotides at internal positions of the sense and / or antisense strands. In some embodiments, the backbone structure is as shown in ND001', ND002', ND009', ND011', ND032', or ND033'.

[0330] In some embodiments, the dsRNA of the present invention further comprises other modified nucleotides. As used herein, “modified nucleotide” refers to a nucleotide other than a ribonucleotide (2'-hydroxynucleotide). In some embodiments, at least 50% (e.g., at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100%) of the nucleotides are modified nucleotides. Exemplary modified nucleosides include, but are not limited to: 2'-O-methyl modified nucleosides, nucleosides containing a 5'-thiophosphate group, terminal nucleosides linked to a cholesterol derivative or a dodecanoic acid diecamide group, locked nucleosides, base-free nucleosides, 2'-deoxyribonucleosides, 2'-fluorinated nucleosides, 2'-amino-modified nucleosides, 2'-alkyl-modified nucleosides, morpholino nucleosides, non-locked nucleosides (UNA), aminophosphates, or nucleosides containing non-natural bases, or any combination thereof.

[0331] In some embodiments, in the dsRNA activator of the present invention, substantially all nucleotides of the sense strand are modified nucleotides; or substantially all nucleotides of the antisense strand are modified nucleotides; or substantially all nucleotides of both the sense strand and the antisense strand are modified nucleotides.

[0332] In some embodiments, all or substantially all nucleotides of the dsRNA activator are modified nucleotides. As described herein, a dsRNA activator in which substantially all nucleotides are modified nucleotides refers to a dsRNA activator having four or fewer (i.e., 0, 1, 2, 3, or 4) nucleotides that are native ribonucleotides in both the sense and antisense strands. As used herein, a sense strand in which substantially all nucleotides are modified nucleotides refers to a sense strand having two or fewer (i.e., 0, 1, or 2) nucleotides that are native ribonucleotides in the sense strand. As used herein, an antisense strand in which substantially all nucleotides are modified nucleotides refers to an antisense strand having two or fewer (i.e., 0, 1, or 2) nucleotides that are native ribonucleotides in the antisense strand.

[0333] In some embodiments, all nucleotides in the sense strand of the dsRNA activator are modified nucleotides and / or all nucleotides in the antisense strand are modified nucleotides; or all nucleotides in both the sense strand and the antisense strand are modified nucleotides.

[0334] In some embodiments, the oligonucleotide in the dsRNA molecule may contain at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least fifteen, at least twenty or more modified nucleosides, or all nucleosides of the oligonucleotide may be modified nucleosides, wherein for each of the multiple modified nucleosides in the dsRNA molecule, the modification is independent and need not be the same.

[0335] In some implementations, the dsRNA sequence (i.e., the nucleoside base sequence) that represses a specific target gene contains different modifications in the phosphate backbone, ribose portion, and / or base portion.

[0336] Examples of nucleoside base modifications that can be used to generate dsRNA activators include the substitution of nucleotides containing uracil, guanine, or adenine with nucleotides containing, for example, inosine; and the replacement of adenine and cytosine in oligonucleotides with guanine and uracil, respectively, to form GU Wobble base pairing with the target mRNA. In addition, other examples of modified nucleoside bases that can be used to generate RNAi activators include, but are not limited to: 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xantine, 4-acetylcytosine, 5-(carboxyhydroxymethyl)uracil, 5-carboxymethylaminomethyl-2-thiouracil, 5-carboxymethylaminomethyluracil, dihydrouracil, β-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7 -Methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, β-D-mannosyl queosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-hydroxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-hydroxyacetic acid methyl ester, uracil-5-hydroxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl)uracil, (acp3)w, and 2,6-diaminopurine. These modified nucleoside bases are all within the scope of this invention.

[0337] Examples of ribosome modifications that can be used to generate dsRNA active agents include ribosome structures modified by replacing one of the following: a hexose ring (HNA), a threonose ring (TNA), a locked nucleic acid (LNA), a bicyclic ring with a bimolecular bridge between the C2 and C4 carbons on the ribosome, or a non-locked nucleic acid (UNA, a ribosome lacking a bond between the C2 and C3 carbons). Examples of usable sugar-modified nucleosides also include, for example, bicyclic hexose nucleic acids (WO 2011 / 017521) or tricyclic nucleic acids (WO 2013 / 154798). Modified nucleosides also include nucleosides in which the sugar portion is replaced by a non-sugar portion, such as in the case of peptide nucleic acids (PNA) or morpholino nucleic acids. Sugar modification also includes modifications by replacing the naturally occurring 2'-OH group on the ribosome ring of the RNA nucleoside with other groups. Furthermore, substituents can be introduced, for example, at the 2', 3', 4', or 5' positions of the sugar ring.

[0338] In some embodiments, the dsRNA activator of the present invention may comprise a 2' sugar-modified nucleotide, such as a 2'-substituted nucleoside. Examples of 2'-substituted modified nucleosides are 2'-O-alkyl-RNA nucleoside, 2'-O-methyl-RNA nucleoside, 2'-alkoxy-RNA nucleoside, 2'-O-methoxyethyl-RNA nucleoside (MOE), 2'-amino-DNA nucleoside, 2'-fluoro-RNA nucleoside, and 2'-F-ANA nucleoside. Other examples may be found, for example, in Freier and Altmann; Nucl. Acid Res., 1997, 25, 4429-4443 and Uhlmann; Curr. Opinion in Drug Development, 2000, 3(2), 293-213 and Deleavey and Damha, Chemistry and Biology 2012, 19, 937. In some embodiments, the RNAi activator according to the present invention comprises at least one 2'-modified nucleotide. In some embodiments, the 2'-modification is selected from 2'-deoxy, 2'-fluoro, 2'-O-methyl, 2'-O-methoxyethyl (2'-O-MOE), 2'-O-allyl, 2'-O-aminopropyl (2'-O-AP), 2'-O-dimethylaminoethyl (2'-O-DMAOE), 2'-O-dimethylaminopropyl (2'-O-DMAP), 2'-O-dimethylaminoethoxyethyl (2'-O-DMAEOE), and 2'-ON-methylacetamido (2'-O-NMA). In some embodiments, the RNAi activator according to the invention comprises at least one 2'-modified nucleoside selected from the following: 2'-O-alkyl-RNA nucleoside, 2'-O-methyl-RNA nucleoside, 2'-alkoxy-RNA nucleoside, 2'-O-methoxyethyl-RNA nucleoside (MOE), 2'-amino-DNA nucleoside, 2'-fluoro-RNA nucleoside, and 2'-F-ANA nucleoside.

[0339] In some embodiments, examples of modifications that may be mentioned, in addition to nucleoside analogs comprising formula (I) of the present invention, may further include, for example: 2'-O-methyl nucleotide modification, 2'-fluoronucleotide modification, 2'-deoxyribonucleotide modification, locked nucleotide (LNA) modification, unlocked nucleotide (UNA) modification, conformation-restricted nucleotide modification, 2'-O-methoxyethyl nucleotide modification, debased nucleotide modification, 2'-amino nucleotide modification, 2'-O-allyl-nucleotide modification, and 2'-C-alkyl-nucleotide modification. 2'-O-alkylnucleotides, morpholinonucleotides, aminophosphamide nucleotide modifications, nucleotide modifications with non-natural bases, tetrahydropyranonucleotide modifications, 1,5-dehydrated hexadiol nucleotide modifications, cyclohexenyl nucleotide modifications, nucleotide modifications containing thiophosphate groups, nucleotide modifications containing methylphosphate groups, nucleotide modifications containing 2'-phosphates, nucleotide modifications containing 5'-phosphates, thermostable nucleotide modifications, ethylene glycol nucleotide (GNA) modifications, and 2-O-(N-methylacetamide) nucleotide modifications; and combinations thereof.

[0340] In some embodiments, the dsRNA activator according to the invention may optionally also comprise a chemical modification at the 5' and / or 3' ends, i.e., a non-nucleotide or nucleoside chemical moiety linked to the end of the oligonucleotide chain (sense and / or antisense strand) of RNAi. Examples of chemical moieties linked to the 3' end of the oligonucleotide chain can be found, for example, in WO 2005 / 021749 and WO 2007 / 128477. Examples of chemical moieties linked to the 5' end of the oligonucleotide chain may include, but are not limited to, 5'-terminal phosphate ester modifications, such as 5'-vinylphosphonate (5'-VP) such as 5'-(E)-vinylphosphonate (5'-(E)-VP), 5'-methylphosphonate (5'-MP), (S)-5'-C-methyl analogues, and 5'-thiophosphate (5'-PS).

[0341] In some embodiments, at least one of the modified nucleotides is selected from the group consisting of: deoxynucleotides, 3'-terminal deoxythymidine (dT) nucleotides, 2'-O-methyl modified nucleotides, 2'-fluorine modified nucleotides, 2'-deoxy modified nucleotides, locked nucleotides, 2'-5'-linked ribonucleotides (3'-RNA), unlocked nucleotides, conformation-restricted nucleotides, restricted ethyl nucleotides, base-free nucleotides, 2'-amino modified nucleotides, 2'-O-allyl modified nucleotides, 2'-C-alkyl modified nucleotides, 2'-methoxyethyl modified nucleotides, 2'- O-alkyl modified nucleotides, morpholinonucleotides, aminophosphates, nucleotides including non-natural bases, tetrahydropyran modified nucleotides, 1,5-dehydrohexyl modified nucleotides, cyclohexenyl modified nucleotides, nucleotides including thiophosphate groups (e.g., nucleosides containing 5'-thiophosphate groups), nucleotides including methylphosphonate groups, nucleotides including 5'-phosphates, nucleotides including 5'-phosphate mimics, vinyl-phosphonate nucleotides, heat-labile nucleotides, ethylene glycol modified nucleotides (GNA), nucleotides including 2'-phosphates and 2-O-(N-methylacetamide) modified nucleotides; and combinations thereof.

[0342] In some embodiments, at least one of the modified nucleotides is selected from the group consisting of: nonlocked nucleotides (UNA), locked nucleotides (LNA), threononucleotides (TNA), CeNA, 2'-methoxyethyl, 2'-O-alkyl, 2'-O-allyl, 2'-C-allyl, 2'-fluoro, 2'-deoxy, and ethylene glycol; and combinations thereof. In some embodiments, at least one of the modified nucleotides in the modified nucleotides of the dsRNA activator is selected from the group consisting of: deoxynucleotides, 2'-O-methyl modified nucleotides, 2'-fluoro modified nucleotides, 2'-deoxy modified nucleotides, nucleotides comprising 2'-phosphate groups, and nucleotides comprising thiophosphate groups; and combinations thereof.

[0343] In some embodiments, the antisense strand of the dsRNA activator of the present invention contains 2'-methoxy (2'-O-methyl) modified nucleotides, for example, all nucleotides are 2'-methoxy modified nucleotides or 1-21 or 1-23 nucleotides are 2'-methoxy modified nucleotides, for example, at least 15, 16, or 17 nucleotides are 2'-methoxy modified nucleotides. In some embodiments, the sense strand of the dsRNA activator of the present invention contains 2'-methoxy (2'-O-methyl) modified nucleotides, for example, all nucleotides are 2'-methoxy modified nucleotides or 1-21 nucleotides are 2'-methoxy modified nucleotides, for example, at least 15 or 16 nucleotides are 2'-methoxy modified nucleotides.

[0344] In some embodiments, the antisense strand of the dsRNA activator of the present invention contains 2'-fluorine modified nucleotides, for example, 1-5 nucleotides are 2'-fluorine modified nucleotides, for example, 4, 5 or 6 nucleotides are 2'-fluorine modified nucleotides. In some embodiments, the sense strand of the dsRNA activator of the present invention contains 2'-fluorine modified nucleotides, for example, 1-5 nucleotides are 2'-fluorine modified nucleotides, for example, 3 or 4 nucleotides are 2'-fluorine modified nucleotides.

[0345] In some embodiments, one or more nucleotides of the dsRNA activator are linked by a non-standard bond or backbone (i.e., a modified nucleotide bond or a modified backbone). In some embodiments, the modified nucleotide bond is a covalent nucleotide bond containing a non-phosphate group. In some embodiments, the modified nucleoside internucleotide bond or skeleton includes, but is not limited to: a 5'-thiophosphate group (represented herein as lowercase "s"), a chiral thiophosphate, a thiophosphate, a dithiophosphate, a triphosphate, an aminoalkyl phosphate triester, an alkylphosphonate (e.g., a methylphosphonate or a 3'-alkylenephosphonate), a chiral phosphonate, a hypophosphonate, a phosphoramide (e.g., a 3'-aminophosphoramide, an aminoalkylphosphoramide, or a thiophosphoramide), a thioalkyl-phosphonate, a thioalkyl phosphate, a morpholino bond, a borophosphate having a normal 3'-5' bond, a borophosphate analog having a 2'-5' bond, or a borophosphate having an antipolarity wherein adjacent nucleoside unit pairs are 3'-5' to 5'-3' or 2'-5'-2' bonds. In some embodiments, the modified nucleoside internucleotide bond or skeleton does not contain a phosphorus atom. In some embodiments, the modified nucleoside interbonds that do not contain phosphorus atoms include, but are not limited to: short-chain alkyl or cycloalkyl sugar interbonds, mixed heteroatom and alkyl or cycloalkyl sugar interbonds, or one or more short-chain heteroatom or heterocyclic sugar interbonds. In some embodiments, the modified nucleoside interskeletons include, but are not limited to: siloxane skeletons, sulfide skeletons, sulfoxide skeletons, sulfone skeletons, formylacetyl and thioformylacetyl skeletons, methyleneformylacetyl and thioformylacetyl skeletons, olefin-containing skeletons, aminosulfonic acid skeletons, methyleneimine and methylenehydrazine skeletons, sulfonate and sulfonamide skeletons, amide skeletons, and other skeletons having mixed N, O, S, and CH2 components.

[0346] In some embodiments, the sense strand of the dsRNA activator may contain 1, 2, 3, 4, 5, or 6 phosphate-thioester bonds. In some embodiments, the antisense strand of the dsRNA activator may contain 1, 2, 3, 4, 5, or 6 phosphate-thioester bonds. In some embodiments, both the sense and antisense strands may independently contain 1, 2, 3, 4, 5, or 6 phosphate-thioester bonds. In some embodiments, the sense strand of the dsRNA activator may contain 1, 2, 3, or 4 phosphate-thioester bonds. In some embodiments, the antisense strand of the dsRNA activator may contain 1, 2, 3, or 4 phosphate-thioester bonds. In some embodiments, both the sense and antisense strands may independently contain 1, 2, 3, or 4 phosphate-thioester bonds.

[0347] In some embodiments, the dsRNA activator's sense strand contains four phosphate-thioester nucleoside bonds. In some embodiments, these two phosphate-thioester nucleoside bonds are located between nucleotides at positions 1-3 starting from the 5' end of the sense strand. In some embodiments, the dsRNA activator's antisense strand contains four phosphate-thioester nucleoside bonds. In some embodiments, these four phosphate-thioester nucleoside bonds are located between nucleotides at positions 1-3 starting from the 5' end and between nucleotides at positions 1-3 starting from the 3' end of the antisense strand. In some embodiments, the dsRNA activator contains four phosphate-thioester nucleoside bonds in both the sense and antisense strands.

[0348] In some embodiments, when the compound of the present invention is added to the 5' or 3' end of the positive strand, the compound is linked to the 5' or 3' nucleotide of the positive strand via a phosphate-thioester nucleoside bond. In this case, the dsRNA activator with the compound of the present invention added to the 5' or 3' end of the positive strand still contains two phosphate-thioester bonds at the 5' and / or 3' ends of the positive strand, but the phosphate-thioester bonds are present between the compound of the present invention at the 5' end of the positive strand and the 1-2 nucleotides counted from the 5' end; and / or the phosphate-thioester bonds are present between the compound of the present invention at the 3' end of the positive strand and the 1-2 nucleotides counted from the 3' end.

[0349] In this document, when referring to the position of the positive or negative chain "starting from the 5' end" or "counting from the 5' end", if the compound of the present invention is added to the 5' end of said chain, it is not counted as the first position. Similarly, when referring to the position of the positive or negative chain "starting from the 3' end" or "counting from the 3' end", if the compound of the present invention is added to the 3' end of said chain, it is not counted as the first position.

[0350] In some embodiments, the 5' end nucleotide of the antisense strand has a 5'-vinyl phosphonate (5'-VP) modification, such as a 5'-(E)-VP modification.

[0351] Other modifications applicable to the dsRNA activator of the present invention can also be found in, for example, the modifications listed in WO2023044094A1, WO2023245060A2, or WO2018 / 027106, the entire contents of which are incorporated herein by reference.

[0352] In some embodiments, the modified nucleotides (5'-3') in the antisense strand of the dsRNA activator of the present invention have the following modification patterns:

[0353] VPNmsNfsNmNmNmNfNmNfNfNmNmNmNmNfNmNfNmNmNmNmNmsNmsNm; or

[0354] VPNmsNfsNmNmNmNfNmNfNfNmNmNmNmNfNmNfNmNmNmsNmsNm; or

[0355] VPNmsNfsNmNmNmNmNfNmNmNmNmNfNmNfNmNfNmNmNmNmsNm; or

[0356] VPNmsNfsNmNmNmNfNmNmNmNmNmNmNmNfNmNfNmNmNmsNmsNm;

[0357] in,

[0358] Nf = any nucleotide modified with 2'-fluorine;

[0359] Nm = any 2'-methoxynucleotide;

[0360] 's' indicates that the two nucleotides are linked by a phosphate thioester bond;

[0361] VP indicates that the 5' end nucleotide of the antisense strand has a 5'-vinyl phosphonate (5'-VP) modification, such as 5'-(E)-VP modification;

[0362] For example, the 5' nucleotide VPNms of the antisense strand can be VPUms, with the following structure:

[0363] In some embodiments, the modified nucleotides (5'-3') in the sense strand of the dsRNA activator of the present invention have the following modification patterns:

[0364] NmsNmsNmNmNm(ND)NfNmNfNfNfNmNmNmNmNmNmNmNmNmsNmsNm, or

[0365] NmsNmsNmNmNm(ND)NfNfNfNmNmNmNmNmNmNmNmNmsNmsNm, or

[0366] (ND)sNmsNmNmNmNmNmNfNmNfNfNfNfNmNmNmNmNmNmNmNmsNmsNm, or

[0367] NmsNmsNmNmNmNmNfNmNfNfNfNmNmNmNmNmNmNmNmNmNmsNms(ND);

[0368] in,

[0369] Nf = any nucleotide modified with 2'-fluorine;

[0370] Nm = any 2'-methoxynucleotide;

[0371] 's' represents a thiophosphate bond;

[0372] (ND) = The form of the compounds of the present invention in a nucleic acid chain, such as the skeletal structure of the compounds of formula (I) or any subform thereof of the present invention, such as the compounds of formula (Ia) to (If) or the specific compounds listed in the examples, such as formula (I”) or any subform thereof.

[0373] In some embodiments, the modified nucleotides (5'-3') in the sense strand of the dsRNA activator of the present invention have the following modification patterns:

[0374] NmsNmsNmNmNm(ND)NfNmNfNfNfNmNmNmNmNmNmNmNmNmsNmsNm, or

[0375] NmsNmsNmNmNm(ND)NfNfNfNmNmNmNmNmNmNmNmNmsNmsNm,

[0376] in,

[0377] Nf = any 2'-fluorine modified nucleotide

[0378] Nm = any 2'-methoxynucleotide;

[0379] 's' represents a thiophosphate bond;

[0380] (ND) = The form of the compounds of the present invention in a nucleic acid chain, such as the skeletal structure of compounds of formula (I) or any subform thereof of the present invention, such as those of formulas (Ia) to (If), or the specific compounds listed in the examples, such as formula (I”) or any subform thereof; wherein the B in the compound is selected from H, a base (e.g., a modified or unmodified base) or -L. B -(C 1-30(Hydrocarbon group), preferably, the compound is selected from ND001, ND002, ND003, ND009, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, ND060, for example selected from ND001, ND002, ND009, ND011, ND032 or ND033, for example, the skeletal structure is such as ND001', N The structures shown in ND002', ND003', ND009', ND011', ND028', ND029', ND030', ND031', ND032', ND033', ND0036', ND042', ND0045', ND0046', ND0047', ND0048', ND052', ND053', ND054', ND055', ND056', ND057', ND058' or ND060' are preferred, with the structures shown in ND001', ND002', ND009', ND011', ND032' or ND033' being preferred.

[0381] In some embodiments, the modified nucleotides (5'-3') in the sense strand of the dsRNA activator of the present invention have the following modification patterns:

[0382] (ND)sNmsNmNmNmNmNmNfNmNfNfNfNfNmNmNmNmNmNmNmNmsNmsNm, or

[0383] NmsNmsNmNmNmNmNfNmNfNfNfNmNmNmNmNmNmNmNmNmNmsNms(ND);

[0384] in,

[0385] Nf = any 2'-fluorine modified nucleotide

[0386] Nm = any 2'-methoxynucleotide;

[0387] 's' represents a thiophosphate bond;

[0388] (ND) = The form of the compounds of the present invention in a nucleic acid chain, such as the skeletal structure of compounds of formula (I) or any subform thereof of the present invention, such as those of formulas (Ia) to (If), or the specific compounds listed in the examples, such as formula (I') or any subform thereof; wherein the B in the compound is selected from H, a base (e.g., a modified or unmodified base) or -L. B -(C1-30 (Hydrocarbon group), preferably, the compound is selected from ND001, ND002, ND003, ND009, ND010, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND036, ND042, ND045, ND046, ND047, ND048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, ND060, for example selected from ND010, ND009, ND036, ND031, ND048 or ND046, for example, the skeletal structure is such as ND001', ND00 The structures shown in ND003', ND009', ND010', ND011', ND028', ND029', ND030', ND031', ND032', ND033', ND0036', ND042', ND0045', ND0046', ND0047', ND0048', ND052', ND053', ND054', ND055', ND056', ND057', ND058' or ND060' are preferred, with the structures shown in ND010', ND009', ND036', ND031', ND048' or ND046' being preferred.

[0389] In some embodiments, the modified nucleotides in the antisense and sense strands of the dsRNA activator of the present invention have the following modification patterns:

[0390] in,

[0391] Nf = any nucleotide modified with 2'-fluorine;

[0392] Nm = any 2'-methoxynucleotide;

[0393] s represents a thiophosphate bond; VP represents a 5'-vinyl phosphonate (5'-VP) modification at the 5' end of the antisense strand, such as 5'-(E)-VP modification;

[0394] (ND) = The form of the compounds of the present invention in a nucleic acid chain, such as the skeletal structure of compounds of formula (I) or any subform thereof of the present invention, such as those of formulas (Ia) to (If), or the specific compounds listed in the examples, such as those of formula (I”).

[0395] For example, the 5' nucleotide VPNms of the antisense strand can be VPUms, with the following structure:

[0396] In some embodiments, the modified nucleotides in the antisense and sense strands of the dsRNA activator of the present invention have the following modification patterns:

[0397] in,

[0398] Nf = any nucleotide modified with 2'-fluorine;

[0399] Nm = any 2'-methoxynucleotide;

[0400] s represents a thiophosphate bond; VP represents a 5'-vinyl phosphonate (5'-VP) modification at the 5' end of the antisense strand, such as 5'-(E)-VP modification;

[0401] (ND) = The form of the compounds of the present invention in a nucleic acid chain, such as the skeletal structure of compounds of formula (I) or any subform thereof of the present invention, such as those of formulas (Ia) to (If), or the specific compounds listed in the examples, such as formula (I”) or any subform thereof; wherein the B in the compound is selected from H, a base (e.g., a modified or unmodified base) or -L. B -(C 1-30 (Hydrocarbon group), preferably, the compound is selected from ND001, ND002, ND003, ND009, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, ND060, for example selected from ND001, ND002, ND009, ND011, ND032 or ND033, for example, the skeletal structure is such as ND001', N The structures shown in ND002', ND003', ND009', ND011', ND028', ND029', ND030', ND031', ND032', ND033', ND0036', ND042', ND0045', ND0046', ND0047', ND0048', ND052', ND053', ND054', ND055', ND056', ND057', ND058' or ND060' are preferred, as shown in ND001', ND002', ND009', ND011', ND032' or ND033'.

[0402] For example, the 5' nucleotide VPNms of the antisense strand can be VPUms, with the following structure:

[0403] In some embodiments, the modified nucleotides in the antisense and sense strands of the dsRNA activator of the present invention have the following modification patterns:

[0404] in,

[0405] Nf = any nucleotide modified with 2'-fluorine;

[0406] Nm = any 2'-methoxynucleotide;

[0407] s represents a thiophosphate bond; VP represents a 5'-vinyl phosphonate (5'-VP) modification at the 5' end of the antisense strand, such as 5'-(E)-VP modification;

[0408] (ND) = The form of the compounds of the present invention in a nucleic acid chain, such as the skeletal structure of compounds of formula (I) or any subform thereof of the present invention, such as those of formulas (Ia) to (If), or the specific compounds listed in the examples, such as formula (I') or any subform thereof; wherein the B in the compound is selected from H, a base (e.g., a modified or unmodified base) or -L. B -(C 1-30(Hydrocarbon group), preferably, the compound is selected from ND001, ND002, ND003, ND009, ND010, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, ND060, for example, the compound is selected from the structure shown in ND009, ND010, ND030, ND031, ND032, ND033, ND036, ND045, ND046, ND047, ND048, for example, the skeleton structure is as shown in ND001', ND002', ND003' The structures shown in ND009', ND010', ND011', ND028', ND029', ND030', ND031', ND032', ND033', ND0036', ND042', ND0045', ND0046', ND0047', ND0048', ND052', ND053', ND054', ND055', ND056', ND057', ND058', or ND060' are preferred. For example, the antisense strand 5' end nucleotide VPNms can be VPUms, with the following structure:

[0409] In some embodiments, the antisense and sense strands of the dsRNA activator of the present invention are as follows:

[0410] in,

[0411] Nf = any nucleotide modified with 2'-fluorine;

[0412] Nm = any 2'-methoxynucleotide;

[0413] 's' represents a thiophosphate bond;

[0414] VP indicates that the 5' end nucleotide of the antisense strand has a 5'-vinyl phosphonate (5'-VP) modification, such as 5'-(E)-VP modification;

[0415] For example, the 5' nucleotide VPNms of the antisense strand can be VPUms, with the following structure:

[0416] The structures of ND001', ND002', ND009', ND010'(ND010'-2), ND011', ND031', ND032', ND033', ND036', ND046' and ND048' are as defined in this document.

[0417] In some embodiments, the present invention provides dsRNA molecules that inhibit SOD1 gene expression via RNA interference. The dsRNA molecules according to the present invention exhibit good SOD1 gene repressive activity and can effectively deliver RNAi to specific tissues or cells, such as muscle tissue (e.g., skeletal muscle tissue or cardiac muscle tissue) or cells, and adipose tissue or cells.

[0418] In some embodiments, the dsRNA molecule of the present invention for inhibiting the SOD1 gene comprises a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises at least 15 consecutive nucleotides, such as 15, 16, 17, 18, 19, 20, or 21, that differ from the nucleotide sequence of 5′-CAUUUUAAUCCUCACUCUAAA-3′ (SEQ ID NO: 3) by no more than 3, for example 3, 2, 1, or 0 nucleotides, and the antisense strand comprises at least 15 consecutive nucleotides, such as 15, 16, 17, 18, 19, 20, 21, 22, or 23, that differ from the nucleotide sequence of 5′-UUUAGAGUGAGGAUUAAAAUGAG-3′ (SEQ ID NO: 1) or 5′-UUUAGAGUGAGGAUUAAAAUG-3′ (SEQ ID NO: 2) by no more than 3, for example 3, 2, 1, or 0 nucleotides.

[0419] The nucleotide at at least one internal position of the positive strand is replaced with the skeletal structure of a compound of formula (I) of the present invention or any subform thereof such as formulas (Ia) to (If) or a specific compound listed in the embodiments, wherein the internal position is an internal position as described herein, for example, not a position within the cleavage site region of the positive strand, for example, the internal position is the 6th position from the 5' end of the positive strand.

[0420] Alternatively, the positive chain may have at least one (preferably one) skeletal structure of a compound of formula (I) or any subform thereof of the present invention, such as formulas (Ia) to (If), added to the 5' end or 3' end;

[0421] Optionally, all nucleotides of the sense strand and the other nucleotides of the antisense strand contain nucleotide modifications selected from the following: 2′-O-methyl modification and 2′-fluorine modification and deoxynucleotide; wherein the sense strand contains at least 2, 3 or 4 2′-fluorine modifications; wherein the antisense strand contains at least 2, 3, 4 or 5 2′-fluorine modifications; wherein the sense strand contains 4 thiophosphate bonds; wherein the antisense strand contains 4 thiophosphate bonds.

[0422] In some embodiments, the dsRNA activator of the present invention, such as siRNA, inhibits the expression of SOD1 (e.g., the human SOD1 gene) by at least about 30%, about 40%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, or about 97%, as determined by, for example, PCR or by a protein-based method (e.g., by immunofluorescence analysis, using, for example, Western blotting or flow cytometry).

[0423] In some embodiments, the SOD1-targeting dsRNA activators of the present invention, such as siRNA (including siRNA with modified nucleotides and siRNA with modified nucleotides and the backbone structure of the compounds of the present invention), are capable of effectively inhibiting the expression of target genes in vivo (e.g., in specific tissues or cells), such as muscle tissue (e.g., skeletal muscle tissue or cardiac tissue) or cells, and adipose tissue or cells.

[0424] In some embodiments, the SOD1-targeting dsRNA activator of the present invention, such as siRNA (including siRNA having modified nucleotides and siRNA having modified nucleotides and the backbone structure of the compounds of the present invention), inhibits SOD1 expression in vivo (e.g., in specific tissues or cells) by at least about 30%, 35%, about 40%, 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%, for example by detection of homogenates of mouse adipose tissue (e.g., subcutaneous fat and / or gonadal fat, or muscle tissue such as skeletal muscle (e.g., quadriceps femoris) or myocardial tissue or cells) extracted after a single intravenous administration to mice, as described in Example 4.

[0425] In some embodiments, the dsRNA activator for inhibiting SOD1 of the present invention comprises a sense strand and an antisense strand, wherein the antisense strand and the sense strand each comprise or consist of the nucleotide sequence shown in SEQ ID NO::

[0426] Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:10;

[0427] Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:11;

[0428] Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:12;

[0429] Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:13;

[0430] Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:14;

[0431] Antisense chain: SEQ ID NO:9; Justice chain: SEQ ID NO:15;

[0432] Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:16;

[0433] Antisense chain: SEQ ID NO:9; Justice chain: SEQ ID NO:17;

[0434] Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:18;

[0435] Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:19;

[0436] Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:20; or

[0437] Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:21.

[0438] In some embodiments, the present invention provides dsRNA molecules that inhibit ACVR1C gene expression via RNA interference. The dsRNA molecules according to the present invention exhibit good ACVR1C gene repressive activity and can efficiently deliver RNAi to specific tissues or cells, such as muscle tissue (e.g., skeletal muscle or cardiac muscle), or cells, as well as adipose tissue or cells.

[0439] In some embodiments, the dsRNA molecule of the present invention for inhibiting the ACVR1C gene comprises a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises at least 15 consecutive nucleotides, such as 15, 16, 17, 18, 19, 20, or 21, differing from the nucleotide sequence of 5′-UGAAAUGCUUGAUGAUACA-3′ (SEQ ID NO: 6) by no more than 3, such as 3, 2, 1, or 0 nucleotides, and the antisense strand comprises at least 15 consecutive nucleotides, such as 15, 16, 17, 18, 19, 20, 21, 22, or 23, differing from the nucleotide sequence of 5′-UGUAUCAUCAAGCAUUUCAGG-3′ (SEQ ID NO: 4) by no more than 3, such as 3, 2, 1, or 0 nucleotides, or...

[0440] The sense strand comprises at least 15 consecutive nucleotides, such as 15, 16, 17, 18, 19, 20, or 21, that differ from the nucleotide sequence of 5′-AAUGCUUGAUGAUACAAUA-3′ (SEQ ID NO: 7) by no more than 3, for example, 3, 2, 1, or 0 nucleotides. The antisense strand comprises at least 15 consecutive nucleotides, such as 15, 16, 17, 18, 19, 20, 21, 22, or 23, that differ from the nucleotide sequence of 5′-UAUUGUAUCAUCAAGCAUUUC-3′ (SEQ ID NO: 5) by no more than 3, for example, 3, 2, 1, or 0 nucleotides.

[0441] The nucleotide at at least one internal position of the positive strand is replaced with the skeletal structure of a compound of formula (I) of the present invention or any subform thereof such as formulas (Ia) to (If) or a specific compound listed in the embodiments, wherein the internal position is an internal position as described herein, for example, not a position within the cleavage site region of the positive strand, for example, the internal position is the 6th position from the 5' end of the positive strand.

[0442] Alternatively, the positive chain may have at least one (preferably one) skeletal structure of a compound of formula (I) or any subform thereof of the present invention, such as formulas (Ia) to (If), added to the 5' end or 3' end;

[0443] Optionally, all nucleotides of the sense strand and the other nucleotides of the antisense strand contain nucleotide modifications selected from the following: 2′-O-methyl modification and 2′-fluorine modification and deoxynucleotide; wherein the sense strand contains at least 2, 3 or 4 2′-fluorine modifications; wherein the antisense strand contains at least 2, 3, 4 or 5 2′-fluorine modifications; wherein the sense strand contains 4 thiophosphate bonds; wherein the antisense strand contains 3 or 4 thiophosphate bonds.

[0444] In some embodiments, the dsRNA active agents of the present invention, such as siRNA, inhibit the expression of ACVR1C (e.g., the human ACVR1C gene) by at least about 30%, about 40%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, or about 97%, as determined by, for example, PCR or by protein-based methods (e.g., by immunofluorescence analysis using, for example, Western blotting or flow cytometry). In some embodiments, the ACVR1C-targeting dsRNA active agents of the present invention, such as siRNA (including siRNA having modified nucleotides and siRNA having modified nucleotides and the backbone structure of the compounds of the present invention), are capable of effectively inhibiting the expression of the target gene in vivo (e.g., in specific tissues or cells), such as muscle tissue (e.g., skeletal muscle tissue or cardiac tissue) or cells, and adipose tissue or cells.

[0445] In some embodiments, the ACVR1C-targeting dsRNA activator of the present invention, such as siRNA (including siRNA having modified nucleotides and siRNA having modified nucleotides and the backbone structure of the compounds of the present invention), inhibits ACVR1C expression in vivo (e.g., in specific tissues or cells) by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%, for example by detection of homogenates of mouse adipose tissue (e.g., subcutaneous fat and / or gonadal fat, or muscle tissue such as skeletal muscle (e.g., quadriceps femoris) or myocardial tissue or cells) extracted after a single intravenous administration to mice, as described in Example 5.

[0446] In some embodiments, the dsRNA activator for inhibiting ACVF1C of the present invention comprises a sense strand and an antisense strand, wherein the antisense strand and the sense strand each comprise or consist of the nucleotide sequence shown in SEQ ID NO::

[0447] Antisense chain: SEQ ID NO:22; Justice chain: SEQ ID NO:25;

[0448] Antisense chain: SEQ ID NO:22; Justice chain: SEQ ID NO:26;

[0449] Antisense chain: SEQ ID NO:23; Justice chain: SEQ ID NO:27; or

[0450] Antisense chain: SEQ ID NO:23; Justice chain: SEQ ID NO:28.

[0451] In some embodiments, the dsRNA activator is prepared or provided in the form of a salt, a mixed salt, or a free acid. In some embodiments, the dsRNA activator is prepared as a sodium salt. Such forms are within the scope of the invention disclosed herein.

[0452] In other embodiments, the dsRNA activator is a pharmaceutically acceptable salt. "Pharmaceutically acceptable salt" as used herein includes, but is not limited to, sodium, calcium, lithium, potassium, ammonium, magnesium, and mixtures thereof. Those skilled in the art will understand that, when provided as a polycationic salt, the dsRNA activator has one cation in each free acid group of the optionally modified phosphodiester backbone and / or any other acidic modification (e.g., a 5'-terminal phosphate group). For example, an oligonucleotide of length "n" nucleotides contains n-1 optionally modified phosphodiester groups, thus an oligonucleotide of length 21 nt can be provided as a salt having up to 20 cations (e.g., 20 sodium cations). Similarly, a dsRNA activator having a 21 nt sense strand and a 23 nt antisense strand can be provided as a salt having up to 42 cations (e.g., 42 sodium cations). In the foregoing examples, if the dsRNA activator further comprises a 5'-terminal phosphate group or a 5'-terminal vinyl phosphate group, the dsRNA activator may be provided as a salt having up to 44 cations (e.g., 44 sodium cations).

[0453] III. Oligonucleotide chains

[0454] In some aspects, the present invention provides an oligonucleotide chain or a pharmaceutically acceptable salt thereof, wherein the oligonucleotide chain comprises, at one or more positions, the skeletal structure of a compound of the present invention or a salt thereof, for example, the skeletal structure of a compound of formula (I) of the present invention or any subform thereof such as formulas (Ia) to (If), or the skeletal structure of a specific compound synthesized in Example 1, wherein the skeletal structure of the compound of formula (I) is shown below:

[0455] The variables are defined as described herein, wherein when the corresponding structure is located at one or more internal positions of the oligonucleotide chain, And ------ are respectively linked to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide; preferably, Linked to the 3' carbon or corresponding position of the previous nucleotide, ------ linked to the 5' carbon or corresponding position of the next nucleotide; or

[0456] When the corresponding structure is located at the 5' end or 3' end of the oligonucleotide chain, One of the two symbols represents a link to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other represents a link to a hydrogen atom, a terminal modification, or a terminal protecting group; preferably, when the corresponding structure is located at the 5' end of the oligonucleotide chain, Linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ linked to the 5' carbon of the 5' terminal nucleotide or the corresponding position; or when the corresponding structure is located at the 3' end of the oligonucleotide chain, ------ linked to hydrogen, a terminal modification, or a terminal protecting group, and Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position.

[0457] In some embodiments, the compound of formula (I) has formula (Ia), and its skeletal structure is shown in the following formula:

[0458] In some embodiments, the compound of formula (I) has a (Ib) structure, and its skeletal structure is shown in the following formula:

[0459] In some embodiments, the compound of formula (I) has the structure of formula (Ic), and its skeletal structure is shown in the following formula:

[0460] In some embodiments, the compound of formula (I) has the structure of formula (Id), and its skeletal structure is shown in the following formula:

[0461] In some embodiments, the compound of formula (I) has the structure of formula (Ie), and its skeletal structure is shown in the following formula:

[0462] In some embodiments, the skeletal structure of the compound of formula (I) is shown in the following formula or any subformulation thereof.

[0463] The variables are defined as described herein, wherein when the corresponding structure is located at one or more internal positions of the oligonucleotide chain, And ------ are respectively linked to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide; preferably, Linked to the 3' carbon or corresponding position of the previous nucleotide, ------ linked to the 5' carbon or corresponding position of the next nucleotide; or when the corresponding structure is located at the 5' end or 3' end of the oligonucleotide chain, One of the two symbols represents a link to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other represents a link to a hydrogen atom, a terminal modification, or a terminal protecting group; preferably, when the corresponding structure is located at the 5' end of the oligonucleotide chain, Linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ linked to the 5' carbon of the 5' terminal nucleotide or the corresponding position; or when the corresponding structure is located at the 3' end of the oligonucleotide chain, ------ linked to hydrogen, a terminal modification, or a terminal protecting group, and Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position.

[0464] In some embodiments, the skeletal structure of the compound of formula (I) is selected from the following structures:

[0465] Wherein, Y is oxygen or sulfur, and where the corresponding structure is located at one or more internal positions of the oligonucleotide chain, And ------ are respectively linked to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide; preferably, Linked to the 3' carbon or corresponding position of the previous nucleotide, ------ linked to the 5' carbon or corresponding position of the next nucleotide; or when the corresponding structure is located at the 5' end or 3' end of the oligonucleotide chain, One of the two symbols represents a link to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other represents a link to a hydrogen atom, a terminal modification, or a terminal protecting group; preferably, when the corresponding structure is located at the 5' end of the oligonucleotide chain, Linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ linked to the 5' carbon of the 5' terminal nucleotide or the corresponding position; or when the corresponding structure is located at the 3' end of the oligonucleotide chain, ------ linked to hydrogen, a terminal modification, or a terminal protecting group, and Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position.

[0466] In some embodiments, the skeletal structure of the compound of formula (I) is selected from the following structures:

[0467] Wherein, when the corresponding structure is located at one or more internal positions of the oligonucleotide chain, And ------ are respectively linked to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide; preferably, Linked to the 3' carbon or corresponding position of the previous nucleotide, ------ linked to the 5' carbon or corresponding position of the next nucleotide; or

[0468] When the corresponding structure is located at the 5' end or 3' end of the oligonucleotide chain, One of the two symbols represents a link to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other represents a link to a hydrogen atom, a terminal modification, or a terminal protecting group; preferably, when the corresponding structure is located at the 5' end of the oligonucleotide chain, Linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ linked to the 5' carbon of the 5' terminal nucleotide or the corresponding position; or when the corresponding structure is located at the 3' end of the oligonucleotide chain, ------ linked to hydrogen, a terminal modification, or a terminal protecting group, and Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position.

[0469] In some embodiments, the oligonucleotide chain of the present invention comprises at least one nucleotide and at least one compound of the present invention. In some embodiments, the oligonucleotide chain of the present invention comprises 15-30 nucleotides (e.g., lengths of about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16 or 15 nucleotides) and at least one, preferably one or two compounds of the present invention.

[0470] In some embodiments, the backbone structure of the compound of the present invention is located at the 5' end and / or 3' end (preferably the 5' end) of the oligonucleotide chain, for example, linked to adjacent nucleotides via phosphate ester bonds or thiophosphate ester bonds. In some specific embodiments, when the backbone structure of the compound of the present invention is located at the 5' end and / or 3' end (preferably the 5' end) of the oligonucleotide chain, the B in the compound is selected from H, a base (e.g., a modified or unmodified base), or -L. B -(C 1-30(Hydrocarbon group). In some more specific embodiments, when the backbone structure of the compound of the present invention is located at the 5' end and / or 3' end (preferably the 5' end) of the oligonucleotide chain, the B in the compound is selected from H or -L. B -(C 1-30 (Hydrocarbon group). In some specific embodiments, the skeletal structure of the compound located at the 5' end and / or 3' end (preferably the 5' end) of the oligonucleotide chain is selected from the skeletal structures of the following compounds: ND001, ND002, ND003, ND009, ND010, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, ND060. In some more specific embodiments, the skeletal structure of the compound located at the 5' end and / or 3' end (preferably the 5' end) of the oligonucleotide chain is preferably selected from the skeletal structures of the following compounds: ND009, ND010, ND030, ND031, ND032, ND033, ND036, ND045, ND046, ND047, and ND048. In some more specific embodiments, the skeletal structure of ND010, ND009, ND036, ND031, ND048, or ND046 may be located at the 5' end and / or 3' end (preferably the 5' end) of the oligonucleotide chain. In some embodiments, the skeleton structures ND001', ND002', ND003', ND009', ND010', ND011', ND028', ND029', ND030', ND031', ND032', ND033', ND0036', ND042', ND0045', ND0046', ND0047', ND0048', ND052', ND053', ND054', ND055', ND056', ND057', ND058', or ND060' are preferably those shown as ND010', ND009', ND036', ND031', ND048', or ND046'.

[0471] In some embodiments, the backbone structure of the compound of the present invention is located at one or more internal positions of the oligonucleotide chain. In some embodiments, the internal positions are selected from any one or more positions other than the three terminal positions at each end of at least one chain. In some embodiments, the internal positions are not located within the cleavage site region of the oligonucleotide chain. In some embodiments, the internal positions are not positions 9-12, 11-13, or 12-14 counting from the 5' end of the positive strand.

[0472] In some embodiments, the oligonucleotide chain is a compound of formula (I) of the present invention at one or more of the following positions: positions 1-8, 6-10, 13-18 or 15-18, for example, positions 1, 5, 6, 7, 15 or 17 on the oligonucleotide chain counting from the 5' end.

[0473] In some implementations, the internal position is the 6th position of the oligonucleotide chain counting from the 5' end.

[0474] In some embodiments, when the backbone structure of the compound of the present invention is located at one or more internal positions of the oligonucleotide chain (e.g., the 6th position counting from the 5' end of the oligonucleotide chain), the B in the compound is preferably H, a base (e.g., a modified or unmodified base) or -L. B -(C 1-30 (hydrocarbon group).

[0475] In some specific embodiments, the backbone structure of the compound located at one or more internal positions of the oligonucleotide chain may be selected from the backbone structures of the following compounds: ND001, ND002, ND003, ND009, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, ND060; preferably selected from the backbone structures of the following compounds: ND001, ND002, ND009, ND011, ND032 or ND033.

[0476] In some more specific embodiments, the backbone structure of ND001, ND002, ND003, ND009, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, or ND060 may be located at one or more internal positions of the oligonucleotide chain. In some embodiments, the skeleton structure is as shown in ND001', ND002', ND003', ND009', ND011', ND028', ND029', ND030', ND031', ND032', ND033', ND0036', ND042', ND0045', ND0046', ND0047', ND0048', ND052', ND053', ND054', ND055', ND056', ND057', ND058', or ND060'.

[0477] In some more specific embodiments, the backbone structure of ND001, ND002, ND009, ND011, ND032, or ND033 may be located at one or more internal positions of the oligonucleotide chain. In some embodiments, the backbone structure is as shown in ND001', ND002', ND009', ND011', ND032', or ND033'.

[0478] In some embodiments, the oligonucleotide chain of the present invention has the same composition as the sense or antisense strand of the dsRNA activator containing the backbone structure of the compound described herein, as defined in Section II herein.

[0479] In some embodiments, the oligonucleotide chain of the present invention has the same composition as the positive strand of the dsRNA activator as defined in Section II herein.

[0480] IV. Definition

[0481] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0482] To explain this specification, the following definitions will be used, and terms used in the singular may also include plural forms, where appropriate. It should be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be restrictive.

[0483] When used in conjunction with a numerical value, the term "about" or "approximately" means to encompass a range of numerical values ​​having a lower limit of 1%, 2%, 3%, 4%, or 5% smaller than the specified numerical value and an upper limit of 1%, 2%, 3%, 4%, or 5% larger than the specified numerical value. It should be understood that the specific value referred to by the term "about" or "approximately" is itself specific and preferably disclosed.

[0484] As used herein, the term “and / or” means any one of the options or two or more of the options.

[0485] As used herein, the terms “comprising” or “including” mean to include the stated elements, integers, or steps, but do not exclude any other elements, integers, or steps. In this document, when the terms “comprising” or “including” are used, unless otherwise specified, they also cover situations consisting of the stated elements, integers, or steps. For example, when referring to a justice chain that “comprising” a specific sequence, it is also intended to cover a justice chain consisting of that specific sequence.

[0486] In this document, "hydrocarbon group" refers to a straight-chain or branched group consisting of carbon and hydrogen atoms having a specified number of carbon atoms, and said group may be saturated or unsaturated. For example, "C 1-30 "Hydrocarbon group" refers to a straight-chain or branched saturated or unsaturated group consisting of 1-30 carbon atoms, including but not limited to alkyl, alkenyl, and alkynyl groups. "C" 1-30 Examples of "hydrocarbon group" include C 12-30 hydrocarbon group, C 14-24 hydrocarbon group, C 16-24 hydrocarbon group, C 16-22 hydrocarbon group, C 18-22 Hydrocarbon groups, etc. Preferably, the "C" 1-30 "Hydrocarbon group" is "C 16-22 "Hydrocarbon group", such as C 16 C 17 C 18 C 19 C 20 C 21 Or C 22Hydrocarbon groups, such as hexadecyl, hexadecylene, hexadecadienyl, heptadecanyl, heptadecanyl, heptadecanyl, octadecyl, octadecylene, octadecadienyl, octadectrienyl, nonadecanyl, nonadecanylene, nonadecanadienyl, nonadecantrienyl, eicosyl, eicosylene, eicosyladienyl, eicosyltrienyl, dodecyl, dodecylene, dodecylene, dodecylene, 6-octyltetradecyl, 10-hexylhexadecyl, all cis 7,10,13,16,19-docosapentaenyl, all cis 4,7,10,13,16,19-docosahexaenyl, all cis Formula 13,16-docosadienyl, all-cis-7,10,13,16-docosatetraenyl, all-cis-4,7,10,13,16-docosapentenyl and cis-13-docosaenyl, such as 1-docoalkyl, 6-octyltetradecane-1-yl, 10-hexylhexadecane-1-yl, cis-docosa-13-en-1-yl, docosane-9-yl, docosane-2-yl, docosane-10-yl, docosane-11-yl and cis-4,7,10,13,16,19-docosahexaen-1-yl, wherein the hydrocarbon group may optionally be substituted with a group selected from the following: hydroxyl, amino, cyano, nitro, halogen, C 1-6 Alkoxy, C 3-8 Cyclic hydrocarbon group, C 3-8 Cycloalkyloxy group, 3-8 membered heterocyclic group, C 1- 6-alkylamino, di(C 1-6 Alkyl) amino, mercapto, azide, alkynyl, aryl (e.g., C) 6-10 aryl) or heteroaryl (such as C) 5-10 (heteroaryl), or optionally, a C12 group is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 Cycloalkyl; preferably, the hydrocarbon group is 1-hexadecyl, 1-eicosyl, 1-monoalkyl or 1-docoalkyl.

[0487] As used herein, the term "halogen" refers to F, Cl, Br, or I.

[0488] As used herein, the term "hydroxyl group" refers to the -OH group.

[0489] As used herein, the term "cyano" refers to the -CN group.

[0490] As used herein, the term "amino" refers to the -NH2 group.

[0491] As used herein, the term "nitro" refers to the -NO2 group.

[0492] As used herein, the term "azido" refers to the -N3 group.

[0493] As used herein, the term "thiol" refers to the -SH group.

[0494] As used herein, the term "alkyl" refers to a saturated aliphatic hydrocarbon group containing a specified number of carbon atoms. For example, "C 1-6 "Alkyl" refers to a saturated aliphatic hydrocarbon group containing 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, etc.

[0495] As used herein, the term "alkenyl" refers to a straight-chain or branched unsaturated hydrocarbon group containing one or more, for example, 1, 2, or 3 carbon-carbon double bonds (C=C), for example, having 2-10 carbon atoms (C=C). 2-10 ), 2-6 carbon atoms (C 2-6 ) or 2-4 carbon atoms (C 2-4 ).

[0496] As used herein, the term "alkynyl" refers to a straight-chain or branched unsaturated hydrocarbon group containing one or more, for example, 1, 2, or 3 carbon-carbon triple bonds (C≡C), such as having 2-10 carbon atoms (C≡C). 2-10 ), more preferably 2-6 carbon atoms (C 2-6 ), more preferably 2-4 carbon atoms (C 2-4 () is a straight-chain or branched unsaturated hydrocarbon group.

[0497] As used herein, the term "alkylene" refers to a fully saturated straight-chain or branched divalent hydrocarbon group consisting of carbon and hydrogen atoms. Alkylenes are, for example, C1... 1- 6-alkylene group, preferably C 1-3 Alkylene, more preferably C 1-2 Alkylenes. Representative examples include, but are not limited to, methylene, ethylene, and propylene. The term "CO alkylene" indicates a valence bond.

[0498] As used herein, the term "alkoxy" refers to an -O-alkyl group, wherein the alkyl group is as defined above.

[0499] The term "cyclic hydrocarbon group" as used in this article refers to a group containing 3-12 cyclic carbon atoms (C 3-12 (For example, 3-10 ring carbon atoms (C) 3-10 ), 3-8 ring carbon atoms (C 3-8 ), 5-7 ring carbon atoms (C 5-7 ), 4-7 ring carbon atoms (C 4-7 ) or 3-6 ring carbon atoms (C 3-6 A saturated or partially unsaturated cyclic hydrocarbon group; it may have one or more rings, for example, 1, 2 or 3, preferably 1 or 2 rings. For example, "C 3-8"Cycloalkyl group" refers to a cycloalkyl group having 3-8 ring carbon atoms. Cycloalkyl groups can include fused or bridged rings and spirocyclic rings. The ring of a cycloalkyl group can be saturated and may contain one or more, for example, one or two double bonds (i.e., partially unsaturated), but it is not fully conjugated and is not an "aryl" group as defined in this invention. Examples of cycloalkyl groups include, for example, C 3-6 Cycloalkyl groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and spiro[2.2]pentyl, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, etc.

[0500] As used herein, the term "heterocyclic group" refers to a saturated or partially unsaturated ring having 3-14 ring atoms (e.g., 4-14 ring atoms (4-14-membered heterocyclic group), 4-12 ring atoms (4-12-membered heterocyclic group), 4-10 ring atoms (4-10-membered heterocyclic group), 3-8 ring atoms (3-8-membered heterocyclic group), 4-8 ring atoms (4-8-membered heterocyclic group), 3-6 ring atoms (3-6-membered heterocyclic group), or 4-5 ring atoms (4-5-membered heterocyclic group)), wherein the ring atoms comprise one or more (e.g., 1, 2, or 3, preferably 1 or 2) heteroatoms independently selected from N, O, and S, and the remaining ring atoms are carbon atoms; it may have one or more rings, such as 1, 2, or 3, preferably 1 or 2 rings. N and S may optionally be oxidized to various oxidation states. For example, "3-8 membered heterocyclic group" means a heterocyclic group having 3-8 (3, 4, 5, 6, 7 or 8) ring atoms, which contains at least one, for example 1, 2 or 3, preferably 1 or 2 heteroatoms independently selected from N, O and S.

[0501] As used herein, the term "aryl" refers to a carbocyclic hydrocarbon group consisting of one or more rings, such as two fused rings, having 6-14 carbon atoms (e.g., 6-14 carbon atoms (6-14-membered aryl), 6-12 carbon atoms (6-12-membered aryl), 6-10 carbon atoms (6-10-membered aryl)), wherein at least one ring is an aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, phenanthryl, indenyl, indenyl, with phenyl and naphthyl being preferred.

[0502] As used herein, the term "heteroaryl" refers to a mono-, di-, or tricyclic ring system having 5-12 ring atoms (e.g., 5-10 ring atoms (5-10-membered heteroaryl), 5-6 ring atoms (5-6-membered heteroaryl), 9-12 ring atoms (9-12-membered heteroaryl)), wherein the ring atoms include one or more, for example 1, 2, 3, or 4, preferably 1, 2, or 3, heteroatoms independently selected from N, O, and S, and the remaining ring atoms are carbon atoms, wherein at least one ring is an aromatic ring, and wherein S and N may optionally be oxidized to various oxidation states.

[0503] As used herein, the term "hydroxyl protecting group" refers to a group that prevents the hydroxyl group from undergoing an undesirable chemical reaction and can be removed under specific conditions to release the hydroxyl group. Examples of such hydroxyl protecting groups include, but are not limited to: trimethylsilyl (TMS), triethylsilyl (TES), dimethylisopropylsilyl (DMIPS), diethylisopropylsilyl (DEIPS), tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), triisopropylsilyl (TIPS), acetyl (Ac), chloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl (TFA), p-methoxybenzoyl, allyloxycarbonyl (Alloc), 2,2,2-trichloroethoxycarbonyl (Troc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), benzyl (Bn), p-methoxybenzyl ( PMB), allyl, triphenylmethyl (Tr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzylmethyl (PMBM), 4,4'-dimethoxytriphenylmethyl (DMTr), monomethoxytriphenylmethyl (MMT), 9-fluorenylmethoxycarbonyl (Fmoc), o-nitrophenylcarbonyl, p-phenylazophenylcarbonyl, benzoyl, p-chlorobenzoyl and 5'-(α-methyl-2-nitropiperyl)oxycarbonyl (MeNPOC), especially 4,4'-dimethoxytriphenylmethyl (DMTr).

[0504] As used herein, a "reactive phosphorus group" refers to a phosphorus-containing group contained in a nucleotide unit or nucleotide analog unit that can react with a hydroxyl or amino group contained in another molecule, particularly in another nucleotide unit or another nucleotide analog, via a nucleophilic attack reaction. Typically, such a reaction produces an ester-type nucleoside bond linking the first nucleotide unit or the first nucleotide analog unit to the second nucleotide unit or the second nucleotide analog unit. The reactive phosphorus group can be selected from activated phosphate ester groups, activated phosphite ester groups, or phosphoramidite groups.

[0505] The compounds of the present invention can be in the form of salts, including acid addition salts and base salts. Preferably, the salt is in the form of a pharmaceutically acceptable salt. "Pharmaceutically acceptable salt" refers to those salts that retain the biological efficacy and properties of the free compound and are not biologically or otherwise undesirable. Acid addition salts can be formed from inorganic or organic acids. Inorganic acid salts include, for example, hydrochlorides, hydrobromates, sulfates, hydrogen sulfates, nitrates, carbonates, phosphates, etc., and organic acid salts include, formates, acetates, trifluoroacetates, propionates, glycolates, gluconates, lactates, pyruvates, oxalates, malates, malonates, glutarate, adipates, succinates, fumarates, maleates, tartrates, citrates, aspartate, sine, ascorbate, glutamate, and ortho- Aminobenzoate, benzoate, cinnamate, mandelate, dihydroxynaphthalate, phenylacetate, methanesulfonate, ethanesulfonate, ethanedisulfonate, benzenesulfonate, p-toluenesulfonate, xylenesulfonate, trimethylbenzenesulfonate (mesitylate), hydroxyethanesulfonate, naphthalenesulfonate, naphthalenedisulfonate, camphorsulfonate, salicylate, oleate, nicotinate, saccharinate, palmitate, stearate, furoate, hippurate, orotate, and pyruvate, etc. Salts also include those derived from inorganic bases, such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts; and those derived from non-toxic organic bases: primary, secondary, and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, tromethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Salts can be synthesized from parent compounds using conventional methods. Pharmaceutically usable salts are preferred. However, other salts can also be useful, for example, in separation or purification steps, and can be used during preparation, and are therefore included within the scope of this disclosure.

[0506] The compounds of this invention may contain one or more asymmetric carbon atoms. Therefore, the compounds may exist as diastereomers, enantiomers, or mixtures thereof. The synthesis of the compounds may employ racemic compounds, diastereomers, or isomers as starting materials or intermediates. A mixture of specific diastereomers can be isolated or enriched with one or more specific diastereomers by chromatographic or crystallographic methods. Similarly, enantiomer mixtures can be isolated or enriched with enantiomers using the same techniques or other techniques known in the art. The asymmetric carbon or nitrogen atom may each be in an R or S configuration, both of which are within the scope of this invention. In the structures shown herein, all stereoisomers are included as compounds of this invention unless the stereochemistry of any particular chiral atom is specified. Stereochemical definitions and conventions used herein follow those commonly used in the art.

[0507] The term "stereoisomer" refers to compounds that have the same chemical composition but differ in the spatial arrangement of their atoms or groups. Stereoisomers include diastereomers, enantiomers, and conformational isomers.

[0508] The term "diastereomer" refers to a stereoisomer that has two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers possess different physical properties, such as melting point, boiling point, spectral properties, and biological activity. Mixtures of diastereomers can be separated using high-resolution analytical techniques such as electrophoresis and chromatographic methods such as HPLC.

[0509] The term "enantiomer" refers to two stereoisomers of a compound that are non-overlapping mirror images of each other.

[0510] The term "tautomer" refers to structural isomers with different energies that can interconvert through low energy barriers. For example, proton tautomers (also known as proton shift tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via the rearrangement of some bonding electrons.

[0511] In the presence of a chiral center, the compounds of the present invention can exist as single enantiomers or mixtures of enantiomers, and those skilled in the art can determine stable and viable isomeric forms of the compounds of the present invention. According to one embodiment, a compound of formula (I) is provided, which is a single enantiomer with an enantiomer excess (%ee) >95%, >98%, or >99%. Preferably, the single enantiomer is present with an enantiomer excess (%ee) >99%.

[0512] As used herein, the term "isotope-labeled compound" refers to a compound in which one or more atoms constituting the compound contain an isotope in a non-natural proportion. The compounds of the present invention may contain an atomic isotope in a non-natural proportion on one or more atoms constituting the compound, thereby forming isotopic variations, whether or not they are radioactive, and are intended to be covered within the scope of the present invention. Unless otherwise indicated, the structural formulas described herein include compounds that differ in the presence of one or more isotope-enriched atoms. Examples of isotopes and pharmaceutically acceptable salts that may be incorporated into the compounds of the present invention include, but are not limited to, isotopes of hydrogen (e.g., ...). 2 H, 3 H); carbon isotopes (e.g., H); 11 C 13 C and 14 C); isotopes of chlorine (e.g.) 36 Cl); isotopes of fluorine (e.g., Cl); 18 F); isotopes of iodine (e.g., F); 123 I and 125 I); nitrogen isotopes (e.g.) 13 N and 15 N); isotopes of oxygen (e.g., N); 15 O、 17 O and 18 O); isotopes of phosphorus (e.g., O); phosphorus isotopes ... 32 P); and isotopes of sulfur (e.g. 35 S). Isotope-labeled compounds (e.g., those labeled with...) 3 H and 14 Those of C) can be used for the analysis of the distribution of compounds or substances in tissues. Tritium (i.e. 3 H) and carbon-14 (i.e. 14 C) Isotopes are useful because of their ease of preparation and detectability. Furthermore, heavier isotopes such as deuterium (i.e., 2 H) substitution can provide some therapeutic benefits arising from increased metabolic stability (e.g., increased in vivo half-life or reduced dose requirement). In some embodiments, in the compounds of the present invention, one or more carbon atoms are enriched. 13 C- or 14 Carbon substitution of C. Isotopes that emit positrons, such as... 15 O、 13 N、 11 C and 18 F can be used in positron emission tomography (PET) studies to detect substrate acceptor occupancy. It should be understood that isotopic variations of the compounds of this invention can generally be prepared using conventional methods and appropriate isotopic variations with suitable reagents.

[0513] Some compounds of the present invention can exist in non-solventized and solvated forms, including hydrated forms. The term "solvent" refers to an association or complex of one or more solvent molecules with a compound of the present invention. Examples of solvents that form solvates include water, isopropanol, ethanol, MeOH, DMSO, EA, acetic acid, and ethanolamine. The term "hydrate" refers to a complex in which the solvent molecule is water. Methods of solvation are well known in the art.

[0514] As used herein, the terms “optional,” “optional,” or “optionally” mean that the event or situation described below may or may not occur, and the description includes both cases where the event or situation occurs and cases where it does not occur. For example, “optionally substituted by…” includes both unsubstituted and substituted by one or more of the described substituents.

[0515] As used herein, a “target sequence” refers to a continuous portion of the nucleotide sequence of an mRNA molecule formed during the transcription of a gene (e.g., the SOD1 gene or ACVR1C), containing the mRNA as a primary transcription product of RNA processing. In some embodiments, the target portion of the sequence will be at least long enough to serve as a substrate for dsRNA-directed cleavage at or near that portion of the nucleotide sequence of the mRNA molecule formed during the transcription of a gene (e.g., the SOD1 gene or ACVR1C). For example, the length of the target sequence can be, for example, 15-36 nucleotides (“nt”), or any sub-length therein. As a non-limiting example, the length of the target sequence can be 15-30 nt, 15-26 nt, 15-23 nt, 15-22 nt, 15-21 nt, 15-20 nt, 15-19 nt, 15-18 nt, 15-17 nt, 18-30 nt, 18-26 nt, 18-23 nt, 18-22 nt, 18-21 nt, 18-20 nt, 18 nt, 19-30 nt, 19-26 nt, The target sequence is 19-23 nucleotides, 19-22 nucleotides, 19-21 nucleotides, 19-20 nucleotides, 19 nucleotides, 20-30 nucleotides, 20-26 nucleotides, 20-25 nucleotides, 20-24 nucleotides, 20-23 nucleotides, 20-22 nucleotides, 20-21 nucleotides, 20 nucleotides, 21-30 nucleotides, 21-26 nucleotides, 21-25 nucleotides, 21-24 nucleotides, 21-23 nucleotides, or 21-22 nucleotides, 21 nucleotides, 22 nucleotides, or 23 nucleotides. In some embodiments of the invention, the target sequence is preferably at least 18, 19, 20, 21, 22, or 23 nucleotides long. In some embodiments of the invention, the target sequence is about 19 to about 23 nucleotides long. In some embodiments of the invention, the target sequence is about 21 nucleotides long. In some embodiments of the invention, the target sequence is about 23 nucleotides long.

[0516] The "SOD1" used in this article refers to superoxide dismutase 1 (SOD1). SOD1 has been found to be closely related to the pathogenesis of amyotrophic lateral sclerosis (ALS) and is an important pathogenic gene for the disease. It was also the first ALS risk gene discovered. Furthermore, mutations in this gene are also found in some familial ALS cases, and these pathogenic mutations mostly occur in exons, leading to dysfunction of the SOD1-encoded protein. In addition, some mutations in SOD1 can make it more prone to Aβ-like aggregation. A few mutant proteins can also, like prions, make normal proteins within the same cell more prone to aggregation. These cells containing abnormal SOD1 aggregation can "infect" other normally functioning cells, causing their intracellular SOD1 to also aggregate.

[0517] The term "ACVR1C" (also known as ALK7) used in this article refers to the type I receptor of the transforming growth factor β (TGF-β) superfamily. The ACVR1C protein contains 493 amino acids, including the catalytic domain of the receptor serine / threonine kinase. Upon ligand binding, ACVR1C phosphorylates the SMAD transcription factor in the cytoplasm, causing it to translocate into the nucleus, where it interacts directly with DNA or forms complexes with other transcription factors, thereby regulating gene expression. Furthermore, ACVR1C is involved in the regulation of cell differentiation and apoptosis. ACVR1C is expressed in various tissues, including adipose tissue (RPKM 19.0) and the colon (RPKM 1.8). ACVR1C is associated with a variety of diseases. Due to its important role in multiple biological processes and its association with various diseases, ACVR1C is a potential therapeutic target.

[0518] The terms "base," "nucleobase," or "nucleoside base" used in this article are interchangeable. Bases are one of the basic building blocks of nucleic acids (DNA and RNA), and are the basic portions of nucleotides. Bases are primarily responsible for the coding and information storage functions of nucleic acids. In DNA and RNA, bases form complementary pairs through hydrogen bonds; this pairing rule is the basis of the double-stranded structure of nucleic acids. The base sequence encodes the genetic information of an organism, forming genes through different arrangements and combinations. The specific structure of bases enables them to interact with specific proteins (such as DNA polymerase and RNA polymerase), participating in biological processes such as DNA replication, transcription, and translation. Bases are mainly divided into two categories: purines and pyrimidines. Natural or unmodified bases generally include guanine, cytosine, adenine, thymine, and uracil. When used herein, the base may also encompass modified bases, such as 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xantine, 4-acetylcytosine, 5-(carboxyhydroxymethyl)uracil, 5-carboxymethylaminomethyl-2-thiouracil, 5-carboxymethylaminomethyluracil, dihydrouracil, β-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, and 7-methylguanine. 5-Methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, β-D-mannosyl Q nucleoside, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-hydroxyacetic acid (v), wybutoxosine, pseudouracil, Q nucleoside (queosine), 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-hydroxyacetic acid methyl ester, uracil-5-hydroxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl)uracil, (acp3)w, and 2,6-diaminopurine.

[0519] “G,” “C,” “A,” “T,” and “U” typically represent nucleotides containing guanine, cytosine, adenine, thymine, and uracil as bases, respectively. Unless otherwise specified, they encompass both native and modified nucleotides. It is understood that the terms “ribonucleotide” or “nucleotide” can also refer to modified nucleotides or surrogate replacement moiety. Those skilled in the art will appreciate that guanine, cytosine, adenine, and uracil can be substituted with other moieties without substantially altering the base-pairing properties of oligonucleotides including nucleotides containing such substitution moieties. For example, but not limited to, nucleotides containing inosine (a nucleoside compound formed by the combination of hypoxanthine and ribose) as a base can pair with nucleotides containing adenine, cytosine, or uracil. Therefore, in the nucleotide sequence of the dsRNA characteristic of this invention, nucleotides containing uracil, guanine, or adenine can be substituted with nucleotides containing, for example, inosine. In another example, adenine and cytosine at any position in the oligonucleotide can be replaced by guanine and uracil, respectively, to form a GU-wobbling base pairing with the target mRNA. Sequences containing such substitution moieties are suitable for the compositions and methods characteristic of this invention.

[0520] When this article refers to a "nucleotide sequence", it means a continuous nucleotide sequence, which can be a natural nucleotide or a modified nucleotide.

[0521] As used interchangeably herein, the terms “dsRNA,” “dsRNA activator,” “double-stranded RNA,” and “double-stranded RNA molecule” refer to a complex of ribonucleic acid molecules having a double-stranded structure comprising two antiparallel and substantially complementary nucleic acid strands having “sense” and “antisense” orientations relative to a target RNA (e.g., SOD1 or ACVR1C). In some embodiments of the invention, the double-stranded RNA (dsRNA) triggers the degradation of the target RNA (e.g., mRNA) through a post-transcriptional gene silencing mechanism referred to herein as RNA interference or RNAi. In some embodiments, the dsRNA of the invention is a small interfering RNA (siRNA). In some embodiments, when referred to herein as a dsRNA activator, it may also comprise a ligand linked to the double-stranded structure, the ligand facilitating the delivery of the dsRNA to a target tissue or target cell.

[0522] The term "siRNA" in this article refers to a class of double-stranded RNA molecules that can mediate the silencing of their complementary target RNA (e.g., mRNA, the transcript of a gene encoding a protein). siRNA is typically double-stranded, consisting of an antisense strand complementary to the target RNA and a sense strand complementary to that antisense strand. For convenience, such mRNA is also referred to herein as the mRNA to be silenced. Such genes are also called target genes. Typically, the RNA to be silenced is an endogenous gene or a pathogen gene.

[0523] The term "antisense strand" or "guide strand" refers to the strand of dsRNA that contains a region that is substantially complementary to the target gene (e.g., SOD1 or ACVR1C mRNA).

[0524] As used herein, the terms “sense strand” or “lackey strand” or “sense strand” refer to an oligonucleotide strand containing a region substantially complementary to the region of an antisense strand as defined herein, which can be complementary to the antisense strand to form a dsRNA, which can be complementary to the antisense strand to form a double-stranded region of the dsRNA.

[0525] In this document, unless otherwise specified, the terms "complementarity" or "complementarity" refer to the ability of an oligonucleotide or polynucleotide containing a first nucleotide sequence to hybridize with an oligonucleotide or polynucleotide containing a second nucleotide sequence under certain conditions and form a double-stranded structure. Those skilled in the art can determine the optimal complementarity of the two sequences and the conditions used to determine this complementarity based on the final application of the hybridized oligonucleotide or polynucleotide. Therefore, in this document, when describing the base pairing between the sense and antisense strands of RNAi or dsRNA, or between the antisense strand and target sequence of RNAi or dsRNA, the terms "complementarity" or "complementarity" should be understood to cover not only 100% complementarity (i.e., complete complementarity) but also less than 100% complementarity (i.e., substantially complementarity), that is, the presence of base mismatches in the complementary double-stranded nucleic acid region that do not substantially affect the RNAi or dsRNA's intended function. As those skilled in the art will appreciate, in double-stranded nucleic acid molecules, when a base on one strand forms a Watson-Crick base pair with a corresponding base on the other strand in a complementary manner, the bases at that position on both strands are considered to be "complementarily paired" or "matched." For example, the purine base adenine (A) is complementary to the pyrimidine base thymine (T) or uracil (U); the purine base guanine (C) is complementary to the pyrimidine base cytosine (G). Correspondingly, a "mismatch" refers to a situation in double-stranded nucleic acids where corresponding bases on one strand are not complementary to each other. However, it should be understood that nucleotides modified in the base portion of RNA nucleosides should also be considered complementary if Watson-Crick base pairing is permitted. Therefore, in this paper, nucleoside base “complementarity” encompasses Watson-Crick base pairing between unmodified and modified nucleobases (see, for example, Hirao et al. (2012) Accounts of Chemical Research, Vol. 45, p. 2055 and Bergstrom (2009) Current Protocols in Nucleic Acid Chemistry Suppl. 37 1.4.1).

[0526] In this document, for the purposes of this invention, the expression "complementary" or "complementarity" associated with double-stranded RNAi activators (such as dsRNAs like siRNA described herein) is preferably not less than 70%, meaning that at least 70% of the base positions in the double-stranded region formed by complementary hybridization are complementary, i.e., the number of mismatched positions in the continuous nucleotide sequence forming the double-stranded region is less than 30%. For example, for a 21-base-pair double-stranded region, not less than 70% complementarity means that the double-stranded region forms no more than 6, 5, 4, 3, 2, 1, or 0 mismatched base pairs during hybridization. Preferably, the presence of insertions and deletions is not allowed when calculating the complementarity of the continuous nucleotide sequence in the double-stranded region. Accordingly, in this document, the expression associated with RNAi activators, "complementary (antisense) sequence" to the target sequence, or "complementary (sense) sequence" to a portion of the antisense sequence, can be "completely complementary" or "substantially complementary." "Completely complementary" means that the two sequences have 100% complementarity. While the first sequence is referred to herein as “substantially complementary” to the second sequence, the ability of the two sequences to hybridize in the resulting duplex may contain one or more, but typically no more than 30%, 20%, or 10%, mismatched base pairs and still remain under conditions most relevant to their final application (e.g., repressing gene expression via a RISC pathway). When used herein, “perfectly complementary” means that the two strands are completely complementary.

[0527] As used herein, “perfect complementarity” means complete complementarity between two strands. It should be understood that when referring to complete complementarity of complementary or double-stranded regions, it means complete complementarity between two identical nucleotide strands that match during alignment. Therefore, it should be understood that when the two oligonucleotides of an RNAi or dsRNA are designed to form one or more single-stranded overhangs during hybridization, such overhangs will not be considered mismatches when determining complementarity. For example, for the purposes described herein, an RNAi containing a 19-nucleotide sense oligonucleotide strand and a 21-nucleotide antisense oligonucleotide strand can still be considered “perfectly complementary” if the longer antisense oligonucleotide contains a 19-nucleotide sequence that is completely complementary to the shorter sense oligonucleotide.

[0528] As used herein, the term "complementary region" refers to a region on the antisense strand that is complementary (substantially complementary or perfectly complementary) to a sequence defined herein (e.g., a target sequence, such as the SOD1 or ACVR1C mRNA target sequence). In cases where the complementary region is not perfectly complementary (substantially complementary) to the target sequence, mismatches can be located within the molecule or in terminal regions. Typically, the most tolerable mismatches are in terminal regions, such as within 5, 4, 3, 2, or 1 nucleotides at the 5' or 3' end of the dsRNA; for example, the first nucleotide at the 5' end of the antisense strand can tolerate a mismatch. In some embodiments, the double-stranded RNA activator of the present invention comprises nucleotide mismatches in the antisense strand. In some embodiments, the antisense strand of the double-stranded RNA activator of the present invention comprises no more than 4 mismatches with the target mRNA; for example, the antisense strand comprises 4, 3, 2, 1, or 0 mismatches with the target mRNA. In some embodiments, the 5' end nucleotide of the antisense strand of the double-stranded RNA activator of the present invention is mismatched with the target mRNA, for example, the 5' end of the antisense strand of the double-stranded RNA activator of the present invention is U, regardless of whether the 3' end of the target mRNA is A, which pairs with U. In some embodiments, the antisense strand of the double-stranded RNA activator of the present invention has no more than four mismatches with the sense strand; for example, the antisense strand contains four, three, two, one, or zero mismatches with the sense strand. In some embodiments, the nucleotide mismatch is, for example, within five, four, or three nucleotides from the 3' end of the antisense strand or the corresponding 5' end of the sense strand. In some embodiments, the nucleotide mismatch is, for example, within five, four, or three nucleotides from the 3' end of the sense strand or the corresponding 5' end of the antisense strand. In another embodiment, the nucleotide mismatch is, for example, at the 3' end nucleotide of the sense or antisense strand.

[0529] Generally, most nucleotides in each strand of a dsRNA molecule are ribonucleotides, but as described in detail herein, each or both strands may also contain one or more non-ribonucleotides, such as deoxyribonucleotides or modified nucleotides. Additionally, as used herein, “dsRNA” can comprise chemically modified ribonucleotides; dsRNA can contain substantial modifications at multiple nucleotide sites. As used herein, the term “modified nucleotide” refers to a nucleotide that independently has a modified sugar moiety, a modified internucleotide bond, or a modified nucleobase, or any combination thereof. Therefore, the term modified nucleotide encompasses substitution, addition, or removal of, for example, functional groups or atoms, of internucleotide bonds, sugar moieties, or nucleobases. Modifications of the active agents suitable for use in this invention include all types of modifications disclosed herein or known in the art.

[0530] As used herein, the term "nucleotide overhang" refers to at least one unpaired nucleotide that protrudes from the double-stranded structure or double-stranded region of dsRNA. A nucleotide overhang exists, for example, when the 3' end of one strand of dsRNA extends beyond the 5' end of the other strand, or vice versa. dsRNA may contain an overhang having at least one nucleotide; alternatively, the overhang may contain at least two, three, four, five, or more nucleotides. The nucleotide overhang may contain or consist of nucleotide / nucleoside analogs (including deoxynucleotides / nucleosides). One or more overhangs may be located on the sense strand, antisense strand, or any combination thereof. Additionally, one or more nucleotides of the overhang may be present at the 5' end, 3' end, or both ends of the antisense strand or sense strand of siRNA. In some embodiments, the overhang is located at the 3' end of the antisense strand, and is, for example, 1, 2, 3, 4, or 5 nucleotides, such as 2 nucleotides.

[0531] "Flat-ended" or "flat-ended" means that there are no unpaired nucleotides at that end of the dsRNA, i.e., no nucleotide overhang. A "flat-ended" dsRNA is a double-stranded dsRNA along its entire length, meaning that there are no nucleotide overhangs at either end of the molecule. The dsRNA of this invention encompasses dsRNAs with flat ends at both the 5' and 3' ends.

[0532] As used herein, the terms “double-stranded region” or “double-stranded body” or “double-stranded body region” are used interchangeably to refer to the double-stranded structure formed by the hybridization of the sense and antisense strands in dsRNA.

[0533] As used herein, the “internal position” of the sense or antisense strand refers to a position other than the positions of the 5’ and 3’ terminal nucleotides of the sense or antisense strand. Therefore, the nucleotide at the “internal position” of the sense or antisense strand is neither a 5’ nor a 3’ nucleotide. In some embodiments, the nucleotide at the “internal position” of the sense or antisense strand is not a nucleotide at the overhang of the sense or antisense strand, but a nucleotide within the double-stranded region (nucleotides at the ends of the non-double-stranded region). For naturally occurring oligonucleotides, the internucleotide bond includes a phosphate ester group that forms a phosphodiester bond between adjacent nucleosides. Herein, the term “modified internucleotide bond” is defined as a bond that covalently links two nucleosides together, other than a phosphodiester (PO) bond. The nucleotide chain of the RNAi according to the invention may contain one or more internucleotide bonds modified from natural phosphodiester bonds. Modified internucleotide bonds contemplated according to the invention include, but are not limited to: thiophosphate bonds, dithiophosphate bonds, methylphosphate bonds, selenophosphate bonds, phosphoramidite bonds, etc. In some embodiments, the nucleoside internucleotides modified in the oligonucleotides used for RNAi in this invention are phosphate thioester bonds.

[0534] When this document refers to replacing or substituting nucleotides at positions within the sense or antisense strand with compounds of the present invention (or the skeletal structure of compounds of the present invention), it means that nucleotides complementary to the nucleotides at positions in the sense strand and corresponding to the nucleotides at positions in the antisense strand, or nucleotides complementary to the nucleotides at positions in the antisense strand and corresponding to the target sequence, are replaced with the skeletal structure of the compounds of the present invention, thereby connecting the skeletal structure of the compounds of the present invention to the two nucleotides adjacent to that position via internucleotide bonds (e.g., phosphate ester bonds or thiophosphate ester bonds).

[0535] When this document refers to adding the compound of the present invention to the 5' or 3' end of the sense or antisense strand, it means that the compound of the present invention is added to the 5' or 3' end of the sense or antisense strand based on the original sequence, so that the compound of the present invention is linked to the 5' nucleotide via a nucleoside bond (e.g., a phosphate ester bond or a thiophosphate bond), or the compound of the present invention is linked to the 3' nucleotide via a nucleoside bond (e.g., a phosphate ester bond or a thiophosphate bond).

[0536] As used herein, the term "ligand moiety" refers to the chemical portion that conjugates to the double helix of dsRNA and can alter the distribution, targeting, or half-life of dsRNA. When "dsRNA" or "dsRNA activator" is used herein, it also encompasses dsRNA containing a ligand moiety unless the context explicitly contradicts this. Similarly, when "siRNA" or "siRNA activator" is used herein, it also encompasses siRNA containing a ligand moiety unless the context explicitly contradicts this.

[0537] As used herein, "the positive and / or antisense chains comprising the skeletal structure of a compound of formula (I) at one or more positions" and similar expressions refer to a structure formed by introducing the skeletal structure of a compound of formula (I) of the present invention at one or more positions (e.g., at the 5' end and / or 3' end) of the positive and / or antisense chains and / or replacing the nucleosides at one or more positions (e.g., internal positions) of the positive and / or antisense chains with the skeletal structure of a compound of formula (I) of the present invention, wherein the skeletal structure of the compound of formula (I) is as follows:

[0538] Wherein X, R3, and B are as defined herein, wherein when the corresponding structures are used to replace nucleotides at one or more internal positions of the sense and / or antisense strands, And ------ are respectively linked to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide; preferably, Linked to the 3' carbon or corresponding position of the previous nucleotide, ------ linked to the 5' carbon or corresponding position of the next nucleotide; or when the corresponding structure is added to the 5' or 3' end of the sense and / or antisense strands. One of the two symbols is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other symbol represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is added to the 5' end of the sense strand and / or antisense strand, Linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ linked to the 5' carbon of the 5' terminal nucleotide or the corresponding position; or when the corresponding structure is added to the 3' end of the sense strand and / or antisense strand, ------ linked to hydrogen, a terminal modification, or a terminal protecting group, and ------ Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position.

[0539] As used herein, the term “suppression” is used interchangeably with “reduction,” “silence,” “downregulation,” “knockdown,” and other similar terms, and includes any level of suppression.

[0540] The expression "inhibit target gene (e.g., SOD1 gene or ACVR1C gene)" is intended to refer to the inhibition of the activity or expression of any target gene (e.g., SOD1 gene or ACVR1C gene). The expression "inhibit target gene (e.g., SOD1 gene or ACVR1C gene) expression" is intended to refer to the inhibition of the expression of any target gene (e.g., SOD1 gene or ACVR1C gene) and variants or mutants of the target gene (e.g., SOD1 gene or ACVR1C gene). Therefore, the target gene (e.g., SOD1 gene or ACVR1C gene) can be a wild-type target gene (e.g., SOD1 gene or ACVR1C gene), a mutant target gene (e.g., SOD1 gene or ACVR1C gene), or a transgenic target gene (e.g., SOD1 gene or ACVR1C gene) in the case of genetically manipulated cells, cell groups, or organisms.

[0541] "Inhibition of target gene (e.g., SOD1 or ACVR1C gene) expression" includes inhibition of any level of target gene (e.g., SOD1 or ACVR1C gene) expression, such as at least partial repression of target gene (e.g., SOD1 or ACVR1C gene) expression. Target gene (e.g., SOD1 or ACVR1C gene) expression can be assessed based on the level or level change of any variable associated with target gene (e.g., SOD1 or ACVR1C gene) expression, such as the mRNA level of the target gene (e.g., SOD1 or ACVR1C gene) or the protein level encoded by the target gene (e.g., SOD1 or ACVR1C gene). This level can be assessed in individual cells or in a group of cells (including, for example, samples derived from an individual). Inhibition can be assessed by a decrease in the absolute or relative level of one or more variables associated with target gene (e.g., SOD1 or ACVR1C gene) expression compared to a control level. The control level can be any type of control level used in the art, such as a baseline level before administration or a level determined from similar untreated or controlled individuals, cells, or samples (e.g., a buffer-only control or an inert control).

[0542] The term "target gene (e.g., SOD1 gene or ACVR1C gene) related disease and / or condition and / or status" as used in this article refers to a disease or condition and / or status caused by or associated with abnormal expression and / or activity of a target gene (e.g., SOD1 gene or ACVR1C gene), including conditions, diseases, disorders and / or symptoms that may benefit from decreased expression, replication or protein activity of the target gene (e.g., SOD1 gene or ACVR1C gene), such as skeletal muscle disorders, cardiomyopathy or adipose tissue disorders.

[0543] The term "effective amount" refers to the amount or dose of the dsRNA active agent or composition or combination of the present invention that produces the intended effect in a patient requiring treatment or prevention after being administered in a single or multiple doses. Depending on the intended effect, it may include "therapeutic effective amount" and "preventive effective amount".

[0544] "Therapeutic effective dose" refers to the amount that, at the required dose and for the required duration, effectively achieves the desired therapeutic outcome. A therapeutic effective dose is also a dose in which any toxic or harmful effects of the dsRNA active agent or composition or combination are less than the beneficial therapeutic effect. Relative to an untreated subject, the "therapeutic effective dose" preferably inhibits a measurable parameter by at least about 30%, and more preferably at least about 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or even 100%. "Prophylactic effective dose" refers to the amount that, at the required dose and for the required duration, effectively achieves the desired preventative outcome. Typically, because prophylactic doses are used in individuals before or at an earlier stage of the disease, the prophylactic effective dose will be less than the therapeutic effective dose.

[0545] The terms “individual” or “subject” may be used interchangeably herein and include mammals. Mammals include, but are not limited to, domesticated animals (e.g., cattle, sheep, cats, dogs, and horses), primates (e.g., human and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). In some embodiments, the individual or subject is a human.

[0546] The term "pharmaceutical excipients" refers to diluents, adjuvants (e.g., Freund's adjuvants (complete and incomplete)), excipients, carriers, or stabilizers that are applied together with the active substance.

[0547] The term "pharmaceutical composition" refers to a composition that is present in a form that allows for the biological activity of the active ingredient contained therein, and that does not contain any additional ingredients that would have unacceptable toxicity to a subject administering the composition. In some embodiments, when referring to "pharmaceutical composition," it also encompasses pharmaceutical preparations formulated as formulations or articles.

[0548] The term "drug combination" refers to a non-fixed or fixed combination, including but not limited to a cassette or pharmaceutical composition. The term "non-fixed combination" means that active ingredients (e.g., (i) the dsRNA active agent of the present invention, and (ii) other therapeutic agents) are administered to a patient simultaneously, without a specific time limit, or sequentially at the same or different time intervals, in separate entities, wherein such administration to the patient provides a preventive or therapeutically effective level. In some embodiments, the dsRNA active agent and other therapeutic agents of the present invention used in the drug combination are administered at levels not exceeding those obtained when used alone. The term "fixed combination" means that two or more active agents are administered to a patient simultaneously in the form of a single entity. Preferably, the dosage and / or time interval of the two or more active agents are selected so that the combined use of the components produces an effect greater than that achieved by using any one component alone in treating a disease or condition. The components in a "drug combination" as defined herein are not limited to being present in the same cassette or pharmaceutical composition, but may be in separate formulations or cassettes. The components may be in separate formulations, which may be the same or different.

[0549] The term "combination therapy" refers to the administration of two or more therapeutic agents or modes of treatment to treat the disease described herein. Such administration includes the co-administration of these therapeutic agents in a substantially simultaneous manner, such as in a single capsule containing active ingredients in a fixed proportion. Alternatively, such administration includes the co-administration of individual active ingredients in multiple or separate containers (e.g., tablets, capsules, powders, and liquids). Powders and / or liquids may be reconstituted or diluted to the desired dose prior to administration. Furthermore, such administration includes the sequential administration of each type of therapeutic agent at substantially the same time or at different times. In either case, the treatment regimen will provide the beneficial effect of the combination of drugs in treating the condition or symptom described herein.

[0550] The term "other therapeutic agents" encompasses any therapeutic agent, other than the dsRNA active agent of the present invention or a pharmaceutical composition comprising it, that is effective for the prevention or treatment of target gene-related diseases and / or conditions (e.g., diseases and / or conditions caused by abnormal expression of the target gene).

[0551] When used in this article, "treatment" means to slow down, interrupt, block, alleviate, stop, reduce, or reverse the progression or severity of existing symptoms, conditions, circumstances, illnesses, or diseases.

[0552] When used in this article, "prevention" includes the suppression of the occurrence or development of a disease or condition or the symptoms of a particular disease or condition.

[0553] V. Pharmaceutical Composition

[0554] This invention also relates to pharmaceutical compositions comprising a dsRNA active agent as described herein or a pharmaceutically acceptable salt thereof. Such pharmaceutical compositions may comprise an effective amount of the dsRNA active agent of this invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically or physiologically acceptable carrier. The carrier is generally selected to suit the intended administration mode and may include features for altering, maintaining, or protecting, for example, the composition's pH, molar osmolality, viscosity, clarity, color, isotonicity, odor, sterility, stability, dissolution or release rate, absorption, or permeability. Typically, these carriers comprise aqueous solutions or alcohol / water solutions, emulsions, or suspensions, including saline and / or buffer media.

[0555] Suitable agents included in a pharmaceutical composition include, but are not limited to, antioxidants (e.g., ascorbic acid, sodium sulfite, or sodium bisulfite), buffers (e.g., borates, bicarbonates, Tris-HCl, citrates, phosphates, or other organic acids), fillers (e.g., mannitol or glycine), chelating agents (e.g., ethylenediaminetetraacetic acid (EDTA)), complexing agents (e.g., polyvinylpyrrolidone, β-cyclodextrin, or hydroxypropyl-β-cyclodextrin), monosaccharides, disaccharides, and other carbohydrates (e.g., glucose, mannose, or dextrin), proteins (e.g., free serum albumin, gelatin, or immunoglobulins), colorants, flavoring agents, emulsifiers, hydrophilic polymers (e.g., polyvinylpyrrolidone), low molecular weight peptides, and salt-forming counterions (e.g., acetones). Preservatives (e.g., sodium), preservatives (e.g., benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenylethanol, methylparaben, propylparaben, chlorhexidine, sorbic acid, or hydrogen peroxide), polyols (e.g., glycerol, propylene glycol, or polyethylene glycol), sugar alcohols (e.g., mannitol or sorbitol), suspending agents, surfactants or wetting agents (e.g., protonylene derivatives; PEG; sorbitol esters; polysorbates, such as polysorbate 20 or polysorbate 80; Triton; tromethamine; lecithin; cholesterol or tyloxapal), stabilizing agents (e.g., sucrose or sorbitol), tension enhancers (e.g., alkali metal halides, such as sodium chloride or potassium chloride or mannitol and sorbitol), and / or pharmaceutical adjuvants. Pharmaceutical compositions can be prepared using conventional excipients known in the art for pharmaceutical products, through any of a variety of techniques (Remington's Pharmaceutical Sciences, 21st edition, University of the Sciences in Philadelphia, Philadelphia, PA, USA (2006)).

[0556] In some embodiments, pharmaceutical compositions comprising the dsRNA active agent described herein may be administered orally to a subject patient in need. When the pharmaceutical compositions are administered orally, they may be formulated as tablets, capsules, granules, powders, or syrups.

[0557] In some preferred embodiments, pharmaceutical compositions comprising the dsRNA active agent described herein can be administered parenterally to a subject in need. When the pharmaceutical compositions are administered parenterally, they can be formulated as intravenous, intramuscular, subcutaneous, or intrathecal injections, or as infusions. Parenterical administration can be performed via subcutaneous, intramuscular, or intravenous injection using a syringe, optionally a pen syringe, or a mechanically driven syringe. Alternatively, parenterical administration can be performed using an infusion pump.

[0558] When considering parenteral administration, pharmaceutical compositions are typically in the form of sterile, pyrogen-free, and parenteral-acceptable compositions. Particularly suitable solvents for parenteral injection are properly preserved sterile isotonic solutions.

[0559] The pharmaceutical composition may be in lyophilized form, such as lyophilized cake.

[0560] The parenteral or oral formulations can be prepared by conventional methods. As needed, the dual dsRNA active agents described herein can be mixed with any conventional additives or excipients, such as binders, disintegrants, lubricants, corrosives, solubilizers, suspending agents, emulsifiers, coating agents, cyclodextrins and / or buffers.

[0561] In some embodiments, the pharmaceutical composition is formulated as a prolonged-release and reservoir formulation to provide prolonged release of the dsRNA active agent described herein. Examples of extended-release formulations for injection can be achieved by including agents that delay absorption (e.g., aluminum monostearate and gelatin) in the pharmaceutical composition. In some embodiments, controlled-release formulations are prepared using biodegradable, biocompatible polymers such as ethylene-vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoesters, and polylactic acid.

[0562] The dosage of the dsRNA active agent and pharmaceutical composition of the present invention can be determined according to the patient's weight, age, sex, disease severity, etc. Subjects can be given therapeutic doses of the pharmaceutical composition such as 0.5 mg / kg, 1 mg / kg, 1.5 mg / kg, 2 mg / kg, 2.5 mg / kg body weight, etc. Dosing frequency can be based on regularity, such as daily, weekly, every two weeks, every three weeks, every one month, every two months, every three months, every four months, every five months, every six months, every seven months, every eight months, every nine months, every ten months, every eleven months, annually, or longer, with repeated administration. After the initial treatment regimen, treatment can be given at a lower frequency; for example, after monthly administration for three months, administration can be continued for six months, one year, or longer. Administration of the pharmaceutical composition may reduce the level of target proteins in, for example, a patient's cells, tissues, blood, urine or other compartments by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% or more.

[0563] In some embodiments, the pharmaceutical composition is in the form of a pharmaceutical formulation. In one embodiment, the pharmaceutical composition comprises the dsRNA active agent of the present invention or a pharmaceutically acceptable salt thereof, and a combination of one or more other therapeutic agents.

[0564] In some embodiments, a medicine or pharmaceutical composition comprising the dsRNA active agent of the present invention or a pharmaceutically acceptable salt thereof having the desired purity can be prepared by mixing it with one or more optional pharmaceutical excipients.

[0565] In some embodiments, the dsRNA activator according to the invention may be present in a non-buffered solution, such as saline or water. In some embodiments, the dsRNA activator is in a buffered solution, such as an acetate, citrate, prolyl, carbonate, or phosphate, or any combination thereof, such as phosphate-buffered saline (PBS).

[0566] The pharmaceutical compositions or formulations of the present invention may further comprise more than one active ingredient, said active ingredient being required for a specific indication to be treated, preferably those active ingredients having complementary activities that do not adversely affect each other. In some embodiments, said other active ingredients are, for example, for the prevention or treatment of diseases and / or conditions related to genes (e.g., SOD1 or ACVR1C genes) targeted by said dsRNA active agents, such as diseases and / or conditions caused by abnormal expression and / or activity of genes (e.g., SOD1 or ACVR1C genes) targeted by said dsRNA active agents, or various therapeutic agents capable of benefiting from inhibiting the expression and / or activity of genes (e.g., SOD1 or ACVR1C genes) targeted by said dsRNA active agents. The active ingredients are suitably combined in amounts effective for the intended use.

[0567] In some embodiments, the pharmaceutical composition or formulation of the present invention may be contained in a vial or in a syringe.

[0568] VI. Drug combinations and pillboxes

[0569] The present invention also provides a pharmaceutical combination comprising the dsRNA activator of the present invention, and one or more other therapeutic agents, said other therapeutic agents being effective for the prevention or treatment of diseases and / or conditions related to genes (e.g., SOD1 or ACVR1C genes) targeted by said dsRNA activator, such as diseases and / or conditions caused by abnormal expression and / or activity of genes (e.g., SOD1 or ACVR1C genes) targeted by said dsRNA activator, or diseases and / or conditions that can benefit from inhibiting the expression and / or activity of genes (e.g., SOD1 or ACVR1C genes) targeted by said dsRNA activator, such as skeletal muscle diseases, cardiomyopathy, or adipose tissue diseases, such as SOD1 or ACVR1C-related diseases and / or conditions.

[0570] Another object of the present invention is to provide a complete pillbox containing the drug combination of the present invention, preferably said pillbox in the form of drug dosage units. This allows dosage units to be provided according to a dosing regimen or drug administration interval.

[0571] In some embodiments, the kit of the present invention comprises, within the same package:

[0572] - A first container containing a pharmaceutical composition comprising the dsRNA active agent of the present invention;

[0573] - A second container containing a pharmaceutical composition comprising other therapeutic agents.

[0574] VII. Uses and Methods

[0575] One aspect of the present invention provides a method for inhibiting the expression and / or activity of a target gene (e.g., the SOD1 gene or the ACVR1C gene) in cells, comprising reacting the cells with a dsRNA activator, pharmaceutical composition, or pharmaceutical combination product of the present invention, thereby inhibiting the expression of the target gene (e.g., the SOD1 gene or the ACVR1C gene) in the cells. In some embodiments, the cells are in a subject. In some embodiments, the subject suffers from a disease and / or condition associated with the target gene (e.g., the SOD1 gene or the ACVR1C gene).

[0576] In some embodiments, contacting the cells with the dsRNA activator inhibits the expression of a target gene (e.g., the SOD1 gene or the ACVR1C gene) by at least about 30%, about 40%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, or about 97%. In some embodiments, inhibiting SOD1 expression reduces the SOD1 protein level in the serum of the subject by at least about 30%, about 40%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99%. In some implementations, administration of the dsRNA activator to a subject results in a reduction in the accumulation of target gene (e.g., SOD1 gene or ACVR1C gene) protein or a decrease in the content of target gene (e.g., SOD1 gene or ACVR1C gene) protein in a specific tissue or cell (e.g., muscle tissue (e.g., skeletal muscle tissue or cardiac muscle tissue) or cell, or adipose tissue or cell) in the subject's body.

[0577] This invention provides, in one aspect, a method for preventing or treating a disease or condition in a subject, comprising administering to the subject an effective amount of the dsRNA activator, pharmaceutical composition, drug combination, or kit of the invention. In some embodiments, the disease or condition is a disease and / or condition related to a gene (e.g., the SOD1 or ACVR1C gene) targeted by the dsRNA activator, such as a disease and / or condition caused by abnormal expression and / or activity of the gene (e.g., the SOD1 or ACVR1C gene) targeted by the dsRNA activator, or a disease and / or condition that can benefit from inhibiting the expression and / or activity of the gene (e.g., the SOD1 or ACVR1C gene), such as musculoskeletal disorders, cardiomyopathy, or adipose tissue disorders, such as SOD1 or ACVR1C-related diseases or conditions. In some embodiments, the disease is a musculoskeletal disorder, cardiomyopathy, or adipose tissue disorder, such as a musculoskeletal disorder, cardiomyopathy, or adipose tissue disorder that can be alleviated, improved, or treated by inhibition of the target gene. In some embodiments, the disease is an SOD1 or ACVR1C-related disease and / or condition.

[0578] In some embodiments, the present invention relates to dsRNA activators, pharmaceutical compositions, drug combinations, or kits for therapeutic purposes, such as for the prevention or treatment of diseases and / or conditions associated with genes (e.g., SOD1 or ACVR1C genes) targeted by the dsRNA activator, such as diseases and / or conditions caused by abnormal expression and / or activity of genes (e.g., SOD1 or ACVR1C genes) targeted by the dsRNA activator, or diseases and / or conditions that can benefit from inhibiting the expression and / or activity of genes (e.g., SOD1 or ACVR1C genes) targeted by the dsRNA activator, such as skeletal muscle disorders, cardiomyopathy, or adipose tissue disorders (e.g., skeletal muscle disorders, cardiomyopathy, or adipose tissue disorders that can be alleviated, improved, or treated via target gene inhibition), such as SOD1 or ACVR1C-related diseases or conditions.

[0579] In some embodiments, the present invention relates to the use of the dsRNA activator, pharmaceutical composition, pharmaceutical combination, or kit of the present invention for the preparation of a medicament, for example for the prevention or treatment of diseases and / or conditions associated with a gene (e.g., SOD1 or ACVR1C gene) targeted by the dsRNA activator, such as diseases and / or conditions caused by abnormal expression and / or activity of the gene (e.g., SOD1 or ACVR1C gene) targeted by the dsRNA activator, or diseases and / or conditions that can benefit from inhibiting the expression and / or activity of the gene (e.g., SOD1 or ACVR1C gene) targeted by the dsRNA activator, such as skeletal muscle diseases, cardiomyopathy, or adipose tissue diseases (e.g., skeletal muscle diseases, cardiomyopathy, or adipose tissue diseases that can be alleviated, improved, or treated by target gene inhibition), such as SOD1 or ACVR1C-related diseases or conditions.

[0580] In some embodiments, the present invention relates to the use of the dsRNA activator, pharmaceutical composition, pharmaceutical combination, or kit of the present invention, for example, for the prevention or treatment of diseases and / or conditions related to genes (e.g., SOD1 or ACVR1C genes) targeted by the dsRNA activator, such as diseases and / or conditions caused by abnormal expression and / or activity of genes (e.g., SOD1 or ACVR1C genes) targeted by the dsRNA activator, or diseases and / or conditions that can benefit from inhibiting the expression and / or activity of genes (e.g., SOD1 or ACVR1C genes) targeted by the dsRNA activator, such as skeletal muscle diseases, cardiomyopathy, or adipose tissue diseases (e.g., skeletal muscle diseases, cardiomyopathy, or adipose tissue diseases that can be alleviated, improved, or treated by target gene inhibition), such as SOD1 or ACVR1C-related diseases or conditions.

[0581] In some embodiments, the disease or condition (e.g., musculoskeletal disease, cardiomyopathy, or adipose tissue disorder) is associated with increased expression or activity of the gene (target gene, such as SOD1 or ACVR1C gene) targeted by the dsRNA activator. In some embodiments, the disease or condition (e.g., musculoskeletal disease, cardiomyopathy, or adipose tissue disorder) is caused by (e.g., aberrant expression) of the targeted gene (target gene, such as SOD1 or ACVR1C gene). In some embodiments, the disease or condition (e.g., musculoskeletal disease, cardiomyopathy, or adipose tissue disorder) is an indication for which one benefits from reduced expression and / or activity of the targeted gene (target gene, such as SOD1 or ACVR1C gene).

[0582] In some embodiments, "increased expression or activity of the gene targeted by the dsRNA activator (target gene, such as SOD1 or ACVR1C gene)" refers to increased expression (e.g., nucleic acid or protein level) or activity of the encoded protein of the target gene (e.g., SOD1 or ACVR1C gene) in a subject-specific tissue or cell, such as muscle tissue (e.g., skeletal muscle tissue or cardiac muscle tissue) or cells, or adipose tissue or cells. In some embodiments, the target gene (e.g., SOD1 or ACVR1C gene) is expressed at moderate or high levels in a subject-specific tissue or cell, such as muscle tissue (e.g., skeletal muscle tissue or cardiac muscle tissue) or cells, or in adipose tissue or cells, in relation to a disease and / or condition associated with the target gene (e.g., SOD1 or ACVR1C gene). In some implementations, the amount of protein encoded by the target gene (e.g., the SOD1 or ACVR1C gene) in a subject-specific tissue or cell, such as muscle tissue (e.g., skeletal muscle tissue or cardiac tissue) or cells, or adipose tissue or cells, associated with a disease and / or condition is higher than the amount of protein encoded by the target gene (e.g., the SOD1 or ACVR1C gene) in the corresponding tissue or cells of a healthy control (e.g., a normal-weight control individual).

[0583] Depending on their therapeutic use, the dsRNA active agent or composition or drug or formulation comprising the present invention may also be administered in combination with one or more other therapies, such as other treatment modalities and / or other therapeutic agents, for the purposes described herein, such as for the prevention and / or treatment of the related diseases or conditions mentioned herein. Therefore, the present invention also relates to combination therapies of the dsRNA active agent or composition or drug or formulation comprising the present invention with one or more other therapies.

[0584] In other respects, the present invention provides the use of the dsRNA activator of the present invention or a composition or drug or formulation thereof in the manufacture or preparation of a drug for the purposes described herein, such as for the prevention or treatment of the related diseases or conditions mentioned herein, diseases and / or conditions related to the gene (e.g., SOD1 or ACVR1C gene) targeted by the dsRNA activator, such as diseases and / or conditions caused by abnormal expression and / or activity of the gene (e.g., SOD1 or ACVR1C gene) targeted by the dsRNA activator, or diseases and / or conditions that can benefit from inhibiting the expression and / or activity of the gene (e.g., SOD1 or ACVR1C gene) targeted by the dsRNA activator, such as skeletal muscle diseases, cardiomyopathy, or adipose tissue diseases (e.g., skeletal muscle diseases, cardiomyopathy, or adipose tissue diseases that can be alleviated, improved, or treated by inhibition of the target gene), such as SOD1 or ACVR1C related diseases or conditions.

[0585] In other respects, the present invention also provides the dsRNA activator of the present invention, or a composition, medicine, formulation, or combination product comprising thereof, for therapeutic purposes, such as for treating the related diseases or conditions mentioned herein, diseases and / or conditions associated with genes (e.g., SOD1 or ACVR1C genes) targeted by the dsRNA activator, such as diseases and / or conditions caused by abnormal expression and / or activity of genes (e.g., SOD1 or ACVR1C genes) targeted by the dsRNA activator, or diseases and / or conditions that can benefit from inhibiting the expression and / or activity of genes (e.g., SOD1 or ACVR1C genes) targeted by the dsRNA activator, such as skeletal muscle diseases, cardiomyopathy, or adipose tissue diseases (e.g., skeletal muscle diseases, cardiomyopathy, or adipose tissue diseases that can be alleviated, improved, or treated by target gene inhibition), such as SOD1 or ACVR1C-related diseases or conditions.

[0586] Subjects may be mammals, such as primates, preferably higher primates, such as humans (e.g., individuals who have the disease described herein or are at risk of having the disease described herein).

[0587] In some implementations, the subject has the disease described herein or is at risk of having the disease described herein.

[0588] The combination therapy of the present invention covers combined administration (e.g., two or more therapeutic agents contained in the same formulation or separate formulations) and separate administration. In the case of separate administration, the dsRNA active agent or composition or drug or formulation of the present invention may be administered before, simultaneously with, and / or after the administration of other therapeutic agents and / or active agents.

[0589] In some implementations, the diseases or conditions described herein are musculoskeletal disorders, cardiomyopathy, or adipose tissue disorders, such as those that can be alleviated, improved, or treated by target gene inhibition.

[0590] In some implementations, the cardiomyopathy is selected from the following: obstructive hypertrophic cardiomyopathy (HOCM); familial hypertrophic cardiomyopathy (FHC); heart failure with preserved ejection fraction (HFPEF); atrial fibrillation (AFIB); ventricular fibrillation (VFIB); angina pectoris; myocardial infarction (MI); heart failure with heart failure or reduced ejection fraction (HFREF); supraventricular tachycardia (SVT); hypertrophic cardiomyopathy (HCM).

[0591] In some implementations, the skeletal muscle disease is selected from the following: myosin-associated hypertrophy, congenital myasthenia gravis syndrome, and facioscapulohumeral muscular dystrophy (FSHD).

[0592] In some implementations, adipose tissue disorders are selected from the following: metabolic disorders, such as metabolic syndrome; carbohydrate disorders, such as type 2 diabetes, prediabetes; lipid metabolism disorders, such as hyperlipidemia, hypertension, cardiovascular disease; and weight disorders (such as overweight, obesity, severe obesity, or extreme obesity, encompassing obesity).

[0593] As used herein, the term "weight disorder" refers to an abnormal or excessive accumulation of fat in an individual that may be detrimental to health, encompassing overweight, obesity, severe obesity, or extreme obesity. Various standards exist for identifying different weight disorder conditions, such as BMI, body fat percentage, waist circumference, and waist-to-hip ratio. The "weight disorder" of this invention encompasses a condition identified as exceeding a normal weight using any of these indicators. In some embodiments, body fat percentage is used to determine weight disorder; for example, for men, a body fat percentage exceeding 25% is considered overweight, while for women, a body fat percentage exceeding 28%, 29%, or 30% is considered overweight. In some embodiments, BMI is used to determine overweight or obesity; for example, a BMI greater than or equal to 23 is considered overweight (Chinese standard), or a BMI greater than or equal to 25 is considered overweight (WHO BMI classification standard).

[0594] Therefore, when referring to the prevention or treatment of diseases and / or conditions related to target genes in this invention, weight management is also included.

[0595] Therefore, the present invention provides a method for weight management in subjects, including administering to the subject an effective amount of the dsRNA active agent, pharmaceutical composition, pharmaceutical combination, or cassette of the present invention.

[0596] The present invention also provides a method for reducing weight or body fat in subjects, including administering to the subjects an effective amount of the dsRNA active agent, pharmaceutical composition, pharmaceutical combination, or cassette of the present invention.

[0597] In some embodiments, the present invention relates to the dsRNA activator, pharmaceutical composition, pharmaceutical combination or kit of the present invention for use in therapies, such as for weight management, weight loss or reduction of body fat.

[0598] In some embodiments, the present invention relates to the use of the dsRNA active agent, pharmaceutical composition, pharmaceutical combination or kit of the present invention for weight management, weight loss or body fat reduction, or for the preparation of a medicament for weight management, weight loss or body fat reduction.

[0599] The dsRNA active agent can be administered to the subject intravenously, subcutaneously, or intramuscularly.

[0600] In some implementations, the dsRNA activator is administered intramuscularly to the subject.

[0601] In some implementations, the dsRNA activator is administered subcutaneously to the subject.

[0602] In some embodiments, the method of the present invention further includes administering to the subject an active agent or therapy suitable for treating or preventing the diseases or conditions mentioned herein, such as musculoskeletal disorders, cardiomyopathy, or adipose tissue disorders, or for weight management.

[0603] In some embodiments, the method or use further includes determining the level of a target gene (e.g., protein or nucleic acid level, such as mRNA level) in a specific tissue or cell, such as muscle tissue (e.g., skeletal muscle tissue or cardiac tissue) or cell, or adipose tissue or cell, in a subject before administering the dsRNA activator. In some embodiments, the level of the target gene is compared with the level of the target gene (e.g., protein or nucleic acid level, such as mRNA level) in a healthy subject (normal weight subject) or in healthy adipose tissue or adipocytes. If the level of the target gene in the subject or in adipose tissue or adipocytes is higher than the level of the target gene in a healthy control, then the subject is administered a dsRNA activator targeting the target gene or a composition, drug, formulation, or combination product containing the target gene. Attached Figure Description

[0604] Figure 1 illustrates the effect of siRNA on SOD1 mRNA expression in Hepa1-6 cells.

[0605] Figures 2A-2F depict the effects of siRNA on SOD1 mRNA expression in different mouse tissues (Figure 2A: gonadal adipose tissue; Figure 2B: subcutaneous adipose tissue; Figure 2C: quadriceps femoris muscle tissue; Figure 2D: heart tissue; Figure 2E: lung tissue; Figure 2F: kidney tissue). Example

[0606] Unless otherwise specified, solvent ratios refer to volume ratios.

[0607] In describing dsRNA molecules in this invention, including in the tables of the above and following embodiments, the following abbreviations are used to denote modified nucleotides, targeting groups, and linking groups. Unless otherwise specified in the sequence, as will be readily understood by those skilled in the art, when present in oligonucleotides, these monomers are interconnected by 5'-3'-phosphodiester bonds:

[0608] A = adenosine-3'-phosphate;

[0609] C = cytidine-3'-phosphate;

[0610] G = Guanosine-3'-phosphate;

[0611] U = uridine-3'-phosphate

[0612] T = 5'-methyluridine-3'-phosphate

[0613] Nf = any 2'-fluorine modified nucleotide

[0614] Nfs = any 2'-fluorine-modified nucleoside-3'-thiophosphate ester

[0615] Af = 2'-Fluoroadenosine-3'-phosphate

[0616] Afs = 2'-Fluoroadenosine-3'-Thiophosphate

[0617] Cf = 2'-Fluorocytidine-3'-phosphate

[0618] Cfs = 2'-Fluorocytidine-3'-Thiophosphate

[0619] Gf = 2'-Fluoroguanosine-3'-phosphate

[0620] Gfs = 2'-Fluoroguanosine-3'-Thiophosphate

[0621] Tf = 2'-Fluoro-5'-methyluridine-3'-phosphate

[0622] Tfs = 2'-Fluoro-5'-methyluridine-3'-thiophosphate

[0623] Uf = 2'-fluorouridine-3'-phosphate

[0624] Ufs = 2'-fluorouridine-3'-thiophosphate

[0625] Nm = any 2'-methoxynucleotide

[0626] Nms = any 2'-methoxynucleoside-3'-thiophosphate

[0627] Am = 2'-methoxyadenosine-3'-phosphate

[0628] Ams = 2'-methoxyadenosine-3'-thiophosphate

[0629] Tm = 2'-methoxythymidine-3'-phosphate

[0630] Tms = 2'-methoxythymidine-3'-thiophosphate

[0631] Um = 2'-methoxyuridine-3'-phosphate

[0632] Ums = 2'-methoxyuridine-3'-thiophosphate

[0633] Gm = 2'-methoxyguanosine-3'-phosphate

[0634] Gms = 2'-methoxyguanosine-3'-thiophosphate

[0635] Cm = 2'-methoxycytidine-3'-phosphate

[0636] Cms = 2'-methoxycytidine-3'-thiophosphate.

[0637] Example 1. Synthesis of a lipid delivery compound

[0638] Example 1.1 Synthesis of compound ND001

[0639] Step 1. 3',5'-Di-O-(tetraisopropyldisiloxy)-1-β-D-arabinuridine (ND001-2)

[0640] At room temperature, 1-β-D-arabinuridine (ND001-1) (20.00 g, 81.94 mmol) was dissolved in anhydrous pyridine (200.0 mL), and 1,3-dichloro-1,1,3,3-tetraisopropyldisiloxane (31.5 mL, 98.33 mmol) was slowly added dropwise at 0 °C. The mixture was then stirred at room temperature for 12 hours. The reaction was quenched with ice water (15.0 mL), and the solvent was removed by concentration under reduced pressure. The residue was dissolved in ethyl acetate (200.0 mL) and washed with saturated sodium bicarbonate solution and brine. The solution was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to remove the solvent and then slurried with acetonitrile (150.0 mL). The suspension was filtered, and the filter cake was washed with acetonitrile and dried to give a white solid powder ND001-2 (25.50 g, 65% yield). 1 H NMR (400MHz, CDCl3) δ10.00(s,1H),7.88(s,1H),6.13(d,J=6.1Hz,1H),5.73(d,J=9.7Hz,1H),4.61(dd,J=8.4 ,6.1Hz,1H),4.23–4.12(m,2H),4.05(dd,J=13.3,2.8Hz,1H),3.80(dt,J=9.0,2.2Hz,1H),1.19–1.03(m,28H). LCMS: m / z=469.2,[M+H] + .

[0641] Step 2.3',5'-Di-O-(tetraisopropyldisiloxy)-2'-O-trifluoromethanesulfonate-1-β-D-arabaruridine (ND001-3)

[0642] ND001-2 (25.20 g, 51.82 mmol) was dissolved in anhydrous dichloromethane (200.0 mL), and 4-dimethylaminopyridine (18.80 g, 155.46 mmol) was added at 0 °C, followed by the slow dropwise addition of trifluoromethanesulfonic anhydride (12.9 mL, 77.73 mmol). The mixture was then brought back to room temperature and stirred for 3 hours. The mixture was quenched with ice water (25.0 mL) and concentrated under reduced pressure to remove the solvent. The residue was dissolved in ethyl acetate (200.0 mL) and washed with saturated ammonium chloride aqueous solution and brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum and purified by rapid column chromatography to give the white solid product ND001-3 (12.02 g, 38% yield). LCMS: m / z = 619.2, [M+H] + .

[0643] Step 3.3',5'-Di-O-(tetraisopropyldisiloxy)-2'-O-phthalimideuridine (ND001-4)

[0644] N-hydroxyphthalimide (12.60 g, 58.20 mmol) was dissolved in anhydrous acetonitrile (100.0 mL) at room temperature, and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (8.7 mL, 58.20 mmol) was slowly added dropwise with stirring. Then, a solution of ND001-3 (12.00 g, 19.40 mmol) in anhydrous acetonitrile (30.0 mL) was slowly added dropwise. After the addition was complete, the reaction was stirred for 2 hours, followed by concentration under reduced pressure to remove the solvent. The residue was dissolved in ethyl acetate (200.0 mL) and extracted successively with sodium bicarbonate and brine. The combined organic phases were dried over anhydrous sodium sulfate and filtered. After concentration under reduced pressure to remove the solvent, the solution was purified by rapid column chromatography to give a white solid powder ND001-4 (11.00 g, 85% yield). 1 H NMR (400MHz, DMSO-d6): δ11.23(s,1H),7.87(s,4H),7.57(d,J=7.9Hz,1H),5.92(s,1H),5.49(d,J=7.9Hz,1H) ,5.02(d,J=4.3Hz,1H),4.37(dd,J=8.7,4.3Hz,1H),4.30-4.22(m,2H),4.04-3.94(m,1H),1.06-0.96(m,28H). LCMS: m / z=632.2,[M+H] + .

[0645] Step 4.3',5'-Di-O-(tetraisopropyldisiloxy)-2'-O-aminouridine (ND001-5)

[0646] At room temperature, ND001-4 (11.00 g, 17.43 mmol) was dissolved in anhydrous methanol (150.0 mL), followed by the addition of an aqueous solution (10 mL) of sodium carbonate (8.72 g, 81.92 mmol) and methylamine hydrochloride (5.56 g, 81.92 mmol). The reaction mixture was stirred for 12 hours after the addition was complete. The reaction solution was concentrated under reduced pressure to remove the organic solvent. Extraction was performed with water (100.0 mL) and ethyl acetate (80.0 mL × 3). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by rapid column chromatography to give the white solid product ND001-5 (5.80 g, 68% yield). LCMS: m / z = 502.2, [M+H] + .

[0647] Step 5.3',5'-Di-O-(tetraisopropyldisiloxy)-2'-O-(dococoylamino)uridine (ND001-6)

[0648] At room temperature, compound ND001-5 (2.40 g, 4.79 mmol) was dissolved in 1,2-dichloroethane (20.0 mL), followed by the addition of docosanoal (2.33 g, 7.18 mmol) and acetic acid (143 mg, 2.39 mmol). After reacting at 60 °C for 12 hours, sodium cyanoborohydride (3.08 g, 47.90 mmol) was added to the reaction solution and the mixture was stirred for 1 hour. The reaction was quenched by adding saturated ammonium chloride aqueous solution under ice-water bath conditions. The mixture was extracted three times with dichloromethane, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give compound ND001-6 (2.50 g, 64% yield) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ11.53(s,1H),9.85(d,J=8.0Hz,1H),7.57(d,J=8.0Hz,1H),5.65(d,J=8.7Hz,1H),5.61(d,J=9.4Hz,1H),5.08(d,J=6.2Hz ,1H),4.64–4.52(m,1H),4.02(t,J=7.1Hz,1H),3.89–3.77(m,2H),3.14– 3.06(m,2H),1.21-1.20(m,40H),1.02-0.95(m,28H),0.84–0.81(m,3H). LCMS: m / z=849.1,[M+H] + .

[0649] Step 6.3',5'-Di-O-(tetraisopropyldisiloxy)-2'-O-(eicosyl(methyl)amino)uridine (ND001-7)

[0650] At room temperature, compound ND001-6 (2.50 g, 3.08 mmol) was dissolved in 1,2-dichloroethane (25.0 mL). Paraformaldehyde (1.85 g, 61.70 mmol) and acetic acid (55 mg, 0.92 mmol) were added to the above solution, and the mixture was heated to 60 °C and stirred overnight. Sodium cyanoborohydride (3.08 g, 47.90 mmol) was added to the reaction solution, and the reaction was carried out at 60 °C for 1 hour. The reaction was quenched by adding saturated ammonium chloride aqueous solution under ice-water bath. The mixture was extracted three times with dichloromethane, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give a colorless oily compound ND001-7 (1.30 g, 51% yield). 1 H NMR (400MHz, DMSO-d6) δ11.36(s,1H),7.66(d,J=8.1Hz,1H),5.67(d,J=1.4Hz,1H),5.53(dd,J=8.1,2.1Hz,1H),4.35(d,J=5.4Hz,1H),4.26( dd,J=9.0,5.4Hz,1H),4.08(d,J=13.3Hz,1H),3.94–3.80(m,2H),2.61(s,3H),1.22-1.20(m,40H),1.05–1.00(m,28H),0.84(d,J=7.0Hz,3H).

[0651] Step 7.2'-O-(eicosyl(methyl)amino)uridine (ND001-8)

[0652] Compound ND001-7 (1.30 g, 1.58 mmol) was dissolved in methanol (15.0 mL), and ammonium fluoride (585 mg, 15.80 mmol) was added. The mixture was reacted at 65 °C for 10 hours. After concentration under reduced pressure, the crude product was purified by column chromatography to give a white solid compound ND001-8 (750 mg, yield 82%). 1H NMR (400MHz, DMSO-d6) δ11.30(s,1H),7.89(d,J=8.1Hz,1H),5.96(d,J=7.3Hz,1H),5.65(d,J=8.1Hz,1H),5.18(d,J=5.2Hz,1H),5.14–5.08(m,1H), 4.27(dd,J=7.3,5.0Hz,1H),4.10(dt,J=6.9,3.2Hz,1H),3.85(q,J=2.9Hz ,1H),3.56(d,J=3.9Hz,2H),2.52(s,3H),1.23(s,40H),0.87–0.82(m,3H). LCMS: m / z=582.4,[M+H] + .

[0653] Step 8.5′-O-(4,4′-dimethoxytriphenylmethyl)-2′-O-(eicosyl(methyl)amino)uridine (ND001-9)

[0654] Compound ND001-8 (750 mg, 1.29 mmol) was dissolved in anhydrous pyridine (7.5 mL) at room temperature. A solution of 4,4′-bismethoxytriphenylmethyl chloride (873 mg, 2.58 mmol) in anhydrous pyridine (7.5 mL) was slowly added, and the reaction was allowed to proceed overnight at room temperature. The reaction was quenched with saturated sodium bicarbonate solution, and the mixture was extracted three times with dichloromethane. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give a white solid compound ND001-9 (500 mg, 44% yield). 1 H NMR(400MHz,DMSO-d6)δ11.36(s,1H),7.64(d,J=8.1Hz,1H),7.39–7.35(m,2H),7.32(d,J=7.3Hz,2H),7 .26–7.21(m,5H),6.92–6.87(m,4H),5.95(d,J=6.4Hz,1H),5.34(d,J=8.0Hz,1H),5.29(d,J=5.6Hz,1H), 4.40–4.33(m,1H),4.19(td,J=5.4,3.3Hz,1H),3.95(q,J=3.5Hz,1H),3.74(s,6H),3.30–3.24(m,1H),3 .16(dd,J=10.7,2.9Hz,1H),2.58–2.54(m,3H),1.39(s,2H),1.20(d,J=15.3Hz,38H),0.87–0.81(m,3H). LCMS: m / z=906.3,[M+H] +.

[0655] Step 9.5′-O-(4,4′-dimethoxytriphenylmethyl)-2′-O-(eicosyl(methyl)amino)uridine-3′-(2-cyanoethyl-N,N-diisopropyl)phosphamide (ND001)

[0656] Compound ND001-9 (500 mg, 0.57 mmol) was dissolved in anhydrous dichloromethane (10.0 mL), and N,N-diisopropylethylamine (0.4 mL, 2.26 mmol) was added. N,N-diisopropylphosphonamide (2-cyanoethyl) ester (201 mg, 0.85 mmol) was added dropwise under an ice bath, and the reaction was carried out at room temperature for 2 hours under a nitrogen atmosphere. The reaction was quenched by adding saturated sodium bicarbonate solution under an ice bath. The mixture was extracted three times with dichloromethane, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give a white solid compound ND001 (370 mg, 60% yield). 1 H NMR(400MHz, DMSO-d6)δ11.40(s,1H),7.68(dd,J=8.1,6.3Hz,1H),7.42–7.34(m,2H),7.33–7.26(m,2H),7.25–7.20(m,4H),6.94– 6.81(m,4H),5.94(s,1H),5.43(d,J=8.0Hz,1H),4.52(dt,J=9.9,5.6Hz,1H),4.38(dt,J=10.4,4.9Hz,1H),4.06(d,J=3.8Hz,1H),3 .82(dd,J=7.9,5.7Hz,1H),3.73(d,J=3.1Hz,6H),3.60–3.47(m,2H),3.24(d,J=6.2Hz,2H),2.80–2.76(m,1H),2.61(d,J=5.9Hz,1 H),2.57–2.50(m,5H),1.41(s,2H),1.20(d,J=11.8Hz,38H),1.11(dd,J=8.7,6.7Hz,9H),0.96(d,J=6.7Hz,3H),0.87–0.81(m,3H). 31 P NMR (162MHz, DMSO-d6) δ 149.65, 149.06. LCMS: m / z=1106.4,[M+Na] + .

[0657] Example 1.2 Synthesis of compound ND002

[0658] 1. Synthesis of intermediate ND002-3

[0659] Step 1. Tert-butyl docosyloxycarbamate (ND002-2)

[0660] Tert-butyl hydroxy(methyl)carbamate (ND002-1) (3.91 g, 23.95 mmol) was dissolved in anhydrous N,N-dimethylformamide (120.0 mL). Sodium hydride (60 wt%, 1.92 g, 47.90 mmol) was added in portions under ice bath conditions. After reacting for 10 minutes, the mixture was moved to room temperature and reacted for 30 minutes. The mixture was then returned to an ice bath, and 1-bromodocosahexadecane (14.00 g, 35.93 mmol) was added. After reacting for 10 minutes, the mixture was moved to room temperature and reacted for another 16 hours. The reaction was quenched with water under ice bath conditions. The mixture was extracted three times with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give a white solid compound ND002-2 (6.98 g, 64% yield).

[0661] Step 2. O-Eocodecanyl-N-methylhydroxylamine (ND002-3)

[0662] Compound ND002-2 (4.60 g, 10.09 mmol) was dissolved in dichloromethane (50.0 mL), and trifluoroacetic acid (5.0 mL) was added. The reaction was carried out at room temperature for 8 hours. Most of the solvent was removed by vacuum concentration, and the mixture was neutralized to alkaline by adding saturated sodium bicarbonate solution. The mixture was extracted three times with dichloromethane, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by column chromatography to give a white solid compound ND002-3 (3.46 g, 96% yield). 1 H NMR (400MHz, CDCl3) δ3.67(t,J=6.8Hz,2H),2.72(s,3H),1.55(p,J=6.8Hz,2H),1.25(s,38H),0.95–0.80(m,3H).

[0663] 2. Synthesis of compound ND002

[0664] Step 1.3',5'-Di-O-(tetraisopropyldisiloxy)-2'-(eicosicoalkoxy(methyl)amino)uridine (ND002-4)

[0665] Anhydrous tetrahydrofuran (5.0 mL) was added to a pressure-resistant tube containing compounds ND001-3 (1.24 g, 2.00 mmol) and ND002-3 (1.42 g, 4.00 mmol), and the reaction was carried out at 100 °C for 40 hours. After concentration under reduced pressure, the crude product was purified by column chromatography to obtain a pale yellow oily compound ND002-4 (410 mg, yield 25%). 1 H NMR (400MHz, CDCl3) δ8.20(s,1H),7.77(d,J=8.1Hz,1H),6.22(s,1H),5.67(d,J= 10.5Hz,1H),4.41(dd,J=9.3,6.8Hz,1H),4.22–4.12(m,2H),3.97(dd,J=13.3,2. 6Hz,1H),3.76(td,J=7.0,3.8Hz,2H),3.15(d,J=8.1Hz,1H),2.84(s,3H),1.63(s ,2H),1.52(q,J=7.0Hz,2H),1.25(s,40H),1.13–1.03(m,24H),0.90–0.86(m,3H).

[0666] Step 2.2'-(Eicosekoxy(methyl)amino)uridine (ND002-5)

[0667] Compound ND002-4 (1.47 g, 1.78 mmol) was dissolved in methanol (18.0 mL), and ammonium fluoride (661 mg, 17.83 mmol) was added. The mixture was reacted at 65 °C for 10 hours. After concentration under reduced pressure, the crude product was purified by column chromatography to give a white solid compound ND002-5 (861 mg, yield 83%). 1 H NMR (400MHz, DMSO-d6) δ11.29(s,1H),7.82(d,J=8.2Hz,1H),6.29(d,J=8.2Hz,1H),5.63(dd,J=8.1,2.2Hz,1H),5.18(d,J=5.5Hz,1H),5.07(t,J=4 .6Hz,1H),4.12(t,J=5.7Hz,1H),3.88–3.74(m,1H),3.60–3.39(m,4H),3 .23(dd,J=8.2,5.3Hz,1H),2.59(s,3H),1.23(s,40H),0.91–0.79(m,3H). LCMS: m / z=582.4,[M+H] + .

[0668] Step 3.5′-O-(4,4′-dimethoxytriphenylmethyl)-2′-(eicosicoalkoxy(methyl)amino)uridine (ND002-6)

[0669] Compound ND002-5 (873 mg, 1.50 mmol) was dissolved in anhydrous pyridine (10.0 mL), and an anhydrous pyridine solution of 4,4′-bismethoxytriphenylmethyl chloride (1.02 g, 3.00 mmol) (5.0 mL) was slowly added. The reaction was allowed to proceed at room temperature for 15 hours. The reaction was quenched with saturated sodium bicarbonate solution, and the mixture was extracted three times with dichloromethane. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give a white solid compound ND002-6 (1.13 g, 85% yield). 1 H NMR (400MHz, DMSO-d6) δ11.33(s,1H),7.57(d,J=8.1Hz,1H),7.38(d,J=7.0Hz,2H),7.31(t,J=7.6Hz ,2H),7.24(d,J=8.8Hz,5H),6.89(d,J=8.8Hz,4H),6.27(d,J=7.2Hz,1H),5.42(d,J=8.1Hz,1H),5.2 5(d,J=5.5Hz,1H),4.11(td,J=5.5,2.9Hz,1H),3.97–3.88(m,1H),3.74(s,6H),3.61–3.43(m,2H),3 .31–3.21(m,2H),3.16(dd,J=10.3,3.9Hz,1H),2.61(s,3H),1.33–1.08(m,40H),0.90–0.78(m,3H).

[0670] Step 4.5′-O-(4,4′-dimethoxytriphenylmethyl)-2′-(eicosicoalkoxy(methyl)amino)uridine-3′-(2-cyanoethyl-N,N-diisopropyl)phosphamide (ND002)

[0671] Compound ND002-6 (865 mg, 0.98 mmol) was dissolved in anhydrous dichloromethane (10.0 mL), and N,N-diisopropylethylamine (0.7 mL, 3.92 mmol) was added. Then, N,N-diisopropylphosphonamide (2-cyanoethyl) ester (0.33 mL, 1.47 mmol) was added dropwise under ice bath conditions. After reacting for 30 minutes, the mixture was allowed to react at room temperature for 3.5 hours. The mixture was then returned to an ice bath, and N,N-diisopropylphosphonamide (2-cyanoethyl) ester (0.22 mL, 0.98 mmol) was added dropwise. After reacting for 30 minutes, the mixture was allowed to react at room temperature for 2 hours. The reaction was quenched with saturated sodium bicarbonate solution under ice bath conditions. The mixture was extracted three times with dichloromethane, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give a white solid compound ND002 (590 mg, 56% yield). 1 H NMR (400MHz, DMSO-d6) δ11.36(s,1H),7.67(d,J=8.1Hz,1H),7.38(t,J=8.1Hz,2H),7.35–7.17(m,7H),6.94–6.7 8(m,4H),6.28(d,J=5.4Hz,1H),5.42(d,J=8.1Hz,0.63H),5.37(d,J=8.1Hz,0.35H),4.41(tt,J=11.3,5.8Hz,1H) ,4.09(dd,J=11.7,5.0Hz,1H),3.80(q,J=6.5Hz,1H),3.73(d,J=2.7Hz,6H),3.69–3.42(m,6H),3.26(d,J=7.5Hz ,2H),2.76(t,J=5.9Hz,1H),2.70–2.55(m,4H),1.38–1.04(m,49H),0.95(d,J=6.7Hz,3H),0.84(t,J=6.7Hz,3H). 31 P NMR (162MHz, DMSO-d6) δ 149.18. LCMS: m / z=1106.4,[M+Na] + .

[0672] Example 1.3 Synthesis of compound ND003

[0673] Step 1.3',5'-Di-O-(tetraisopropyldisiloxy)-2'-O-(eicosanoylamino)uridine (ND003-1)

[0674] Docosanoic acid (408 mg, 1.20 mmol) was dissolved in anhydrous dichloromethane (20.0 mL), and N,N-diisopropylethylamine (0.43 mL, 2.49 mmol) and 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (455 mg, 1.20 mmol) were added at 0 °C. Then, a solution of ND001-5 (500 mg, 1.00 mmol) in dichloromethane (5.0 mL) was slowly added, and the reaction was carried out at room temperature for 6 hours. The reaction solution was washed with saturated sodium bicarbonate aqueous solution and brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum and purified by rapid column chromatography to obtain the target product ND003-1 (800 mg, 96% yield). 1 H NMR (400MHz, DMSO-d6) δ11.38(s,1H),11.09(s,1H),7.66(d,J=8.1Hz,1H),5.88(s,1H),5.55(d,J=8.1Hz,1H),4.48(d,J=5.2Hz,1H),4.43–4.32( m,1H),4.17–4.08(m,1H),4.00–3.87(m,2H),2.03–1.90(m,2H),1.57–1. 42(m,2H),1.35–1.15(m,36H),1.13–0.93(m,28H),0.85(t,J=6.8Hz,3H). LCMS: m / z=824.0,[M+H] + .

[0675] Step 2.2'-O-(eicosanoylamino)uridine (ND003-2)

[0676] ND003-1 (500 mg, 0.60 mmol) was dissolved in anhydrous methanol (5.0 mL) at room temperature, followed by the addition of ammonium fluoride (290 mg, 6.80 mmol), and the mixture was heated to 60 °C for 4 hours. The mixture was concentrated under reduced pressure to remove the solvent and purified by rapid column chromatography to obtain the target product ND003-2 (300 mg, yield 67%). 1H NMR (400MHz, DMSO-d6): δ11.50(s,1H),11.41(s,1H),7.92(d,J=8.0Hz,1H), 6.02(d,J=6.8Hz,1H),5.78–5.67(m,2H),5.23(t,J=4.8Hz,1H),4.38(td,J=6 .8,2.0Hz,1H),4.07(s,1H),3.96(d,J=1.6Hz,1H),3.66–3.50(m,2H),1.99(t ,J=7.2Hz,2H),1.57–1.42(m,2H),1.33–1.12(m,36H),0.85(t,J=6.8Hz,3H). LCMS: m / z = 581.9, [M+H] + .

[0677] Step 3.5′-O-(4,4′-dimethoxytriphenylmethyl)-2′-O-(eicosanoylamino)uridine (ND003-3)

[0678] At room temperature, ND003-2 (830 mg, 1.43 mmol) was dissolved in anhydrous pyridine (8.0 mL). Under nitrogen protection, a solution of 4,4′-bismethoxytriphenylmethyl chloride (725 mg, 2.14 mmol) in anhydrous pyridine (2.0 mL) was slowly added dropwise, followed by stirring at room temperature overnight. The reaction mixture was quenched with methanol (5.0 mL) and concentrated under reduced pressure to remove pyridine. The mixture was washed with saturated sodium bicarbonate aqueous solution and brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum and purified by rapid column chromatography to obtain the target product ND003-3 (650 mg, yield 53%). 1 H NMR (400MHz, DMSO-d6) δ11.56(s,1H),11.42(s,1H),7.65(d,J=8.1Hz,1H),7.36–7.20(m,9H),6.8 9(d,J=8.8Hz,4H),6.00(d,J=5.9Hz,1H),5.75(s,1H),5.36(d,J=8.1Hz,1H),4.54(t,J=5.4Hz,1H) ,4.22(d,J=4.4Hz,1H),4.02(q,J=3.8Hz,1H),3.73(s,6H),3.19(dd,J=10.7,3.1Hz,1H),3.02(d,J =7.2Hz,1H),2.02(t,J=7.4Hz,2H),1.56–1.42(m,2H),1.20(d,J=21.5Hz,36H),0.86–0.82(m,3H). LCMS: m / z=884.0,[M+H] + .

[0679] Step 4.5′-O-(4,4′-dimethoxytriphenylmethyl)-2′-(eicosanoylamino)uridine-3′-(2-cyanoethyl-N,N-diisopropyl)phosphamide (ND003)

[0680] Compound ND003 was prepared from compound ND003-3 using a method similar to that described in Example 1.1.

[0681] Example 1.4 Synthesis of compound ND009

[0682] Step 1. 1,2-O-(isopropylidene)-3,5-di-O-(phenylmethyl)-α-D-xylfuranose (ND009-2)

[0683] At room temperature, ND009-1 (10.00 g, 52.58 mmol) was dissolved in anhydrous tetrahydrofuran (100.0 mL), and sodium hydride (5.05 g, 210.32 mmol) was added to the above system, and the mixture was stirred for 1 hour. Benzyl bromide (35.97 g, 210.32 mmol) was added to the above system at 0 °C, and the reaction system was then heated to room temperature and stirred for 12 hours. The reaction system was quenched with ice water and extracted with dichloromethane. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to obtain a yellow oil ND009-2 (18.50 g, 95% yield). 1 H NMR (400MHz, DMSO-d6) δ7.41–7.25(m,10H),5.76–5.73(m,1H),4.71(t,J=4.1Hz,1H),4.66(d,J=11.9Hz,1H),4.50(q,J=6.3Hz,2H),4.45(d,J=1 2.1Hz,1H),4.06–3.99(m,1H),3.76(dd,J=9.2,4.3Hz,1H),3.67(dd,J=11.2,2.1Hz,1H),3.50(dd,J=11.2,5.1Hz,1H),1.47(s,3H),1.31(s,3H).

[0684] Step 2. 1-Deoxy-3,5-di-O-(phenylmethyl)-α-D-xylfuranose (ND009-3)

[0685] ND009-2 (7.00 g, 18.91 mmol) was dissolved in anhydrous dichloromethane (70.0 mL), and trimethylsilyl trifluoromethanesulfonate (21.01 g, 94.55 mmol) was added at 0 °C, followed by triethylsilane (11.00 g, 94.55 mmol). The reaction mixture was brought to room temperature and stirred for 3 hours. The reaction was quenched with saturated sodium bicarbonate solution, extracted with dichloromethane, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum and purified by rapid column chromatography to give a colorless oil, ND009-3 (3.90 g, 66% yield). 1 H NMR(400MHz, DMSO-d6)δ7.43–7.15(m,10H),4.71(d,J=12.0Hz,1H),4.56–4.44(m,3H),4.24(q,J=4.4Hz,1H),3 .96–3.85(m,2H),3.77(dd,J=6.8,4.7Hz,1H),3.59(ddd,J=24.0,9.9,3.3Hz,2H),3.47(dd,J=10.7,5.2Hz,1H).

[0686] Step 3. 1-Deoxy-2-O-(eicosyl)-3,5-di-O-(benzyl)-α-D-xylfuranose (ND009-4)

[0687] At room temperature, ND009-3 (2.88 g, 9.17 mmol) was dissolved in anhydrous N,N-dimethylformamide (30.0 mL), and sodium hydride (0.88 g, 36.67 mmol) was slowly added with stirring. The reaction mixture was stirred for one hour, followed by the slow addition of 1-bromodocosahexadecane (5.36 g, 13.75 mmol) and 18-crown ether-6 (0.24 g, 0.92 mmol). The reaction mixture was stirred for 12 hours, quenched with ice water, and extracted with dichloromethane. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum and purified by rapid column chromatography to give a white solid ND009-4 (4.07 g, 71% yield). 1 H NMR (400MHz, CDCl3) δ7.36–7.27(m,10H),4.66(d,J=11.9Hz,1H),4.54(m,3H),4.15–4.09(m,1H),4.05–3.97(m,1H), 3.94–3.84(m,3H),3.67–3.59(m,1H),3.55–3.40(m,3H),1.60(d,J=7.1Hz,2H),1.26(s,38H),0.88(t,J=6.8Hz,3H).

[0688] Step 4. 1-Deoxy-2-O-(dococoyl)-α-D-xylanose (ND009-5)

[0689] At room temperature, ND009-4 (3.85 g, 6.18 mmol) was dissolved in anhydrous tetrahydrofuran (50.0 mL), followed by the addition of Pd / C (1.50 g, 14.10 mmol). The reaction mixture was heated at 60 °C. ℃ The mixture was stirred for 12 hours under a hydrogen atmosphere. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent. The crude product was purified by rapid column chromatography to give a white solid product ND009-5 (2.37 g, yield 87%). 1 H NMR(400MHz, CDCl3) δ4.05(dd,J=9.7,5.0Hz,2H),3.94–3.89(m,1H),3.89–3.81(m,2H),3.79(dt,J=6.7,3.6Hz,1H),3.68 (dd,J=11.8,4.1Hz,1H),3.61–3.53(m,1H),3.52–3.44(m,1H),1.63(d,J=6.6Hz,2H),1.26(m,38H),0.88(t,J=6.8Hz,3H). LCMS: m / z=443.4,[M+H] + .

[0690] Step 5. 5-O-(4,4′-dimethoxytriphenylmethyl)-1-deoxy-2-O-(dococoyl)-α-D-xylfuranose (ND009-6)

[0691] Compound ND009-5 (2.37 g, 5.36 mmol) was dissolved in anhydrous pyridine (30.0 mL) at room temperature, and 4,4′-bismethoxytriphenylmethyl chloride (2.00 g, 5.89 mmol) was slowly added. The reaction was allowed to proceed for 12 hours at room temperature. The reaction was quenched with saturated sodium bicarbonate solution, and the mixture was extracted three times with dichloromethane. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the organic solvent. The crude product was purified by column chromatography to give compound ND009-6 (2.91 g, 73% yield) as a white solid. 1H NMR(400MHz,DMSO-d6)δ7.39(d,J=7.6Hz,2H),7.33–7.18(m,7H),6.91–6.83(m,4H),4.64 (d,J=6.5Hz,1H),3.92(dd,J=9.2,4.9Hz,1H),3.84(t,J=5.7Hz,1H),3.79(dt,J=8.7,4.0 Hz,2H),3.73(s,6H),3.65(dd,J=9.2,4.0Hz,1H),3.53(s,1H),3.43–3.38(m,1H),3.07(d ,J=3.0Hz,1H),2.97–2.89(m,1H),1.49(d,J=7.0Hz,2H),1.23(s,38H),0.90–0.81(m,3H).

[0692] Step 6. 5-O-(4,4′-dimethoxytriphenylmethyl)-1-deoxy-2-O-(eicosyl)-α-D-xylanose-3-(2-cyanoethyl-N,N-diisopropyl)phosphamide (ND009)

[0693] Compound ND009-6 (378 mg, 0.51 mmol) was dissolved in anhydrous dichloromethane (5.0 mL), and 4,5-dicyanimidazole (DCI) (48 mg, 0.41 mmol) was added. 2-cyanoethyl-N,N,N′,N′-tetraisopropylphosphonamide (P-reagent) (184 mg, 0.61 mmol) was added dropwise at 0 °C, and the reaction was carried out at room temperature for 2 hours under a nitrogen atmosphere. The reaction was quenched by adding saturated sodium bicarbonate solution in an ice bath. The mixture was extracted three times with dichloromethane, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give a white solid compound ND009 (290 mg, 61% yield). 1H NMR (400MHz, DMSO-d6) δ7.42–7.35(m,2H),7.29–7.22(m,6H),7.18(t,J=7.2Hz,1H),6.85(s ,4H),4.11(ddt,J=21.4,10.8,5.2Hz,1H),3.94(td,J=9.2,4.5Hz,3H),3.71(s,9H),3.60–3 .35(m,5H),3.25–3.11(m,1H),2.94(d,J=5.1Hz,1H),2.70(p,J=5.4Hz,1H),1.48(d,J=7.0H z,2H),1.22(d,J=4.9Hz,38H),1.11–1.02(m,9H),0.91(d,J=6.7Hz,3H),0.85–0.80(m,3H). 31 P NMR (162MHz, DMSO-d6) δ 148.33 (d, J=9.5Hz).

[0694] Example 1.5 Synthesis of compound ND010

[0695] Step 1. 1-O-methyl-2-deoxy-5-O-triphenylmethyl-D-ribose (ND010-2)

[0696] Compound 1-O-methyl-2-deoxy-D-ribose (ND010-1, 20.00 g, 135.00 mmol) was dissolved in anhydrous pyridine (200.0 mL). Triphenylchloromethane (45.20 g, 162.00 mmol) was added in portions under ice bath conditions. After reacting for 10 minutes, the mixture was moved to room temperature and reacted for 18 hours. After the reaction was completed, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give a colorless oily compound ND010-2 (35.00 g, 66% yield). 1 H NMR (400MHz, CDCl3) δ7.57–7.46(m,5H),7.41–7.20(m,10H),5.12–5.04(m,1H),4.49–4.36(m,1H),4.05–3.96(m, 1H),3.41–3.36(m,1H),3.32(s,3H),3.29–3.18(m,1H),2.32–2.16(m,1H),2.13–2.06(m,1H),1.38–1.26(m,1H).

[0697] Step 2. 1-O-methyl-2-deoxy-3-O-dococoyl-5-O-triphenylmethyl-D-ribose (ND010-3)

[0698] Compound ND010-2 (35.00 g, 89.70 mmol) was dissolved in N,N-dimethylformamide (350.0 mL). Sodium hydride (60 wt%, 7.17 g, 179.40 mmol) was added in portions under ice bath conditions. After reacting for 30 minutes, the mixture was brought to room temperature, and 1-bromodocosahexadecane (52.30 g, 134.50 mmol) was added. The reaction was allowed to proceed for 8 hours. After the reaction was completed, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give a colorless oily compound ND010-3 (33.00 g, 53% yield).

[0699] Step 3. 1,2-Dideoxy-3-O-dococoyl-D-ribose (ND010-4)

[0700] Compound ND010-3 (10.00 g, 14.30 mmol) was dissolved in anhydrous dichloromethane (100.0 mL). Triethylsilane (6.8 mL, 42.90 mmol) and boron trifluoride diethyl ether complex (11.0 mL, 42.9 mmol) were slowly added dropwise under ice bath conditions, and the reaction was carried out at 0 °C for 5 hours. After the reaction was completed, potassium carbonate (14.46 g, 104.70 mmol) was added and stirred for 30 minutes. Then, the reaction was quenched with saturated sodium bicarbonate solution. The mixture was extracted three times with dichloromethane, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give a white solid compound ND010-4 (3.50 g, yield 57%). 1 H NMR (400MHz, CDCl3) δ4.06–3.88(m,4H),3.84–3.74(m,1H),3.67–3.58(m,1H),3.53–3 .38(m,2H),2.11–1.96(m,2H),1.77(s,1H),1.37–1.23(m,40H),0.92(t,J=6.7Hz,3H).

[0701] Step 4. 2-Cyanoethyl[((2R,3S)-3-(eicosicoalkoxy)tetrahydrofuran-2-yl)methyl]diisopropylphosphonamide (ND010)

[0702] Compound ND010-4 (3.30 g, 7.73 mmol) was dissolved in anhydrous dichloromethane (66.0 mL). N,N-diisopropylethylamine (5.38 mL, 30.92 mmol) and N,N-diisopropylphosphonamide (2-cyanoethyl) ester (2.58 mL, 11.59 mmol) were slowly added dropwise under ice bath conditions. The reaction was carried out at 0 °C for 4 hours. After the reaction was complete, the reaction was quenched with saturated sodium bicarbonate solution. The mixture was extracted three times with dichloromethane, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give a white solid compound ND010 (1.40 g, 29% yield). 1 H NMR (400MHz, CDCl3) δ4.04–3.70(m,6H),3.69–3.55(m,2H),3.45(t,J=6.5Hz,2H),2.68(t,J= 6.5Hz,2H),2.08–1.95(m,2H),1.60(t,J=7.1Hz,2H),1.39–1.15(m,52H),0.98–0.86(m,3H). 31 P NMR (162MHz, CDCl3) δ149.71. LCMS: m / z=627.7,[M+H] + .

[0703] Example 1.6 Synthesis of compound ND011

[0704] Step 1. 3',5'-Di-O-(tetraisopropyldisiloxy)-2'-O-(N-methyleneamino)uridine (ND011-1)

[0705] ND001-5 (7.90 g, 15.75 mmol) was dissolved in 1,2-dichloroethane (80.0 mL) at room temperature, followed by the addition of acetic acid (946 mg, 15.75 mmol) and paraformaldehyde (615 mg, 20.47 mmol). The reaction mixture was then heated to 60 °C and reacted for 4 hours. After the reaction solution was returned to room temperature, it was concentrated under reduced pressure to remove the organic solvent. The product ND011-1 (7.5 g, 93% yield) was purified by column chromatography. LCMS: m / z = 514.0, [M+H] + .

[0706] Step 2. 3',5'-Di-O-(tetraisopropyldisiloxy)-2'-O-(N-methylamino)uridine (ND011-2)

[0707] ND011-1 (7.61 g, 14.80 mmol) was dissolved in anhydrous methanol (100.0 mL), and pyridine 4-methylbenzenesulfonic acid (18.60 g, 74.00 mmol) was added at 0 °C, followed by slow addition of sodium cyanoborohydride (1.91 g, 29.92 mmol). After stirring for 20 minutes, the mixture was brought back to room temperature and stirred for 2 hours. The mixture was quenched with saturated ammonium chloride (25.0 mL) and concentrated under reduced pressure to remove the solvent. The residue was dissolved in dichloromethane (60.0 mL) and washed with saturated sodium bicarbonate aqueous solution and brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum and purified by rapid column chromatography to give the target product ND011-2 (2.40 g, 31% yield). 1 H NMR (400MHz, CDCl3) δ8.68(s,1H),7.71(s,1H),5.87(s,1H),5.67(d,J=8.1Hz,1H),4.31–4.23(m,2H),4.19(dd,J=13.3,1.4Hz,1H), 4.05(d,J=9.0Hz,1H),3.97(dd,J=13.4,2.6Hz,1H),2.80(s,3H),1.31(d,J=19.4Hz,4H),1.10–1.02(m,24H).LCMS: m / z=515.9,[M+H] + .

[0708] Step 3. 3',5'-Di-O-(tetraisopropyldisiloxy)-2'-O-(N-methyleicosanoylamino)uridine (ND011-3)

[0709] Docosanoic acid (1.60 g, 4.70 mmol) was dissolved in anhydrous dichloromethane (20.0 mL), and N,N-diisopropylethylamine (2.2 mL, 12.80 mmol) and 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (2.45 g, 4.70 mmol) were added at 0 °C. Then, a solution of ND011-2 (2.20 g, 4.27 mmol) in dichloromethane (10.0 mL) was slowly added, and the mixture was refluxed at 60 °C for 40 hours. The reaction solution was washed with saturated sodium bicarbonate aqueous solution and brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum and purified by rapid column chromatography to obtain the target product ND011-3 (3.20 g, 91% yield). 1H NMR (400MHz, DMSO-d6) δ11.43(s,1H),7.69(d,J=8.1Hz,1H),5.88(s,1H),5.55(d,J=10 .1Hz,1H),4.74(d,J=5.1Hz,1H),4.35(dd,J=9.7,4.8Hz,1H),4.21(d,J=13.5Hz,1H),4. 04(d,J=9.4Hz,1H),3.96(d,J=11.4Hz,1H),3.26(s,3H),2.41–2.18(m,2H),1.56-1.42( m,6H),1.25(s,36H),1.11–0.96(m,24H),0.86(d,J=7.0Hz,3H).LCMS: m / z=838.9,[M+H] + .

[0710] Step 4. 2'-O-(N-methyleicosanoylamino)uridine (ND011-4)

[0711] ND011-3 (3.20 g, 3.82 mmol) was dissolved in anhydrous methanol (35.0 mL) at room temperature, followed by the addition of ammonium fluoride (1.40 g, 3.82 mmol), and the mixture was heated to 60 °C for 4 hours. The mixture was concentrated under reduced pressure to remove the solvent and purified by rapid column chromatography to obtain the target product ND011-4 (1.70 g, 75% yield). LCMS: m / z = 596.0, [M+H] + .

[0712] Step 5. 5′-O-(4,4′-dimethoxytriphenylmethyl)-2′-O-(N-methyleicosanoylamino)uridine (ND011-5)

[0713] At room temperature, ND011-4 (1.71 g, 2.87 mmol) was dissolved in anhydrous pyridine (20.0 mL), and a solution of 4,4′-bismethoxytriphenylmethyl chloride (1.94 g, 5.72 mmol) in anhydrous pyridine (5.0 mL) was slowly added dropwise under nitrogen protection, followed by stirring at room temperature overnight. The reaction mixture was quenched with methanol (10.0 mL) and concentrated under reduced pressure to remove pyridine. The mixture was washed with saturated sodium bicarbonate aqueous solution and brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum and purified by rapid column chromatography to obtain the target product ND011-5 (2.15 g, 84% yield). 1H NMR(400MHz, DMSO-d6)δ11.48(s,1H),7.74(d,J=8.1Hz,1H),7.44–7.33(m,4H),7.32–7.25(m,5H),6.94 (d,J=9.0Hz,4H),6.09(d,J=5.0Hz,1H),5.82(d,J=5.8Hz,1H),5.39(d,J=8.1Hz,1H),4.64(t,J=5.3Hz, 1H),4.34(s,1H),4.09(d,J=3.2Hz,1H),3.78(s,6H),3.45–3.38(m,1H),3.28(s,1H),3.24(s,3H),2.41 –2.18(m,2H)1.49(t,J=6.9Hz,2H),1.26(t,J=10.8Hz,36H),0.93–0.84(m,3H).LCMS: m / z=898.0,[M+H] + .

[0714] Step 6. 5′-O-(4,4′-dimethoxytriphenylmethyl)-2′-O-(N-methyleicosanoylamino)uridine-3′-(2-cyanoethyl-N,N-diisopropyl)phosphamide (ND011)

[0715] Compound ND011-5 (500 mg, 0.56 mmol) was dissolved in anhydrous dichloromethane (3.0 mL), and 4,5-dicyanimidazole (53 mg, 0.45 mmol) was added. N,N,N,N-tetraisopropylphosphonamide (2-cyanoethyl) ester (0.7 mL, 2.21 mmol) was added dropwise under an ice bath, and the reaction was carried out at room temperature for 16 hours under a nitrogen atmosphere. The reaction was quenched by adding saturated sodium bicarbonate solution under an ice bath. The mixture was extracted three times with dichloromethane, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to obtain the target product ND011 (240 mg, 40% yield). 1H NMR (400MHz, DMSO-d6) δ11.42(s,1H),7.75(d,J=8.1Hz,1H),7.41–7.23(m,9H),6.92–6.86(m,4H),6.04(d,J=4.9 Hz,1H),5.33(dd,J=8.0,1.6Hz,1H),4.80(t,J=4.9Hz,1H),4.51(dt,J=9.5,4.9Hz,1H),4.23(q,J=3.9Hz,1H),3.7 3(s,6H),3.71–3.50(m,4H),3.37(t,J=4.3Hz,2H),3.11(s,3H),2.60(t,J=5.9Hz,2H),2.41–2.18(m,2H),1.43(t, J=6.6Hz, 2H), 1.20 (d, J=12.0Hz, 36H), 1.12 (dd, J=9.3, 6.7Hz, 12H), 0.87–0.80 (m, 3H). LCMS: m / z=1120.0, [M+Na] + .

[0716] Example 1.7 Synthesis of compound ND028

[0717] Step 1. 3',5'-Di-O-(tetraisopropyldisiloxy)-2'-O-(hexadecylamino)uridine (ND028-1)

[0718] At room temperature, compound ND001-5 (4.21 g, 8.40 mmol) was dissolved in 1,2-dichloroethane (25.0 mL), followed by the addition of hexadecane (3.03 g, 12.60 mmol) and acetic acid (250 mg, 4.20 mmol). After reacting at 60 °C for 12 hours, sodium cyanoborohydride (10.56 g, 168.00 mmol) was added to the reaction solution, and the reaction was continued at 60 °C with stirring for another 12 hours. The reaction was quenched by adding saturated ammonium chloride aqueous solution in an ice-water bath. The mixture was extracted three times with dichloromethane, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give a white solid compound ND028-1 (1.51 g, 25% yield). 1H NMR (400MHz, DMSO-d6) δ11.56(d,J=7.9Hz,1H),9.80(d,J=51.3Hz,1H),7.63(dd,J= 8.1,2.1Hz,1H),5.73–5.60(m,2H),5.10(dd,J=22.1,6.0Hz,1H),4.61(ddd,J=17.4 ,9.4,5.9Hz,1H),4.08(dt,J=13.6,3.5Hz,1H),3.92(dd,J=16.1,5.2Hz,1H),3.88– 3.78(m,1H),3.12(s,2H),1.26(s,28H),1.12–1.02(m,28H),0.88(t,J=6.7Hz,3H). LCMS: m / z=726.5,[M+H] + .

[0719] Step 2. 3',5'-Di-O-(tetraisopropyldisiloxy)-2'-O-(hexadecyl(methyl)amino)uridine (ND028-2)

[0720] At room temperature, compound ND028-1 (1.51 g, 2.08 mmol), paraformaldehyde (1.24 g, 41.63 mmol), and acetic acid (37.3 mg, 0.62 mmol) were dissolved in 20.0 mL of 1,2-dichloroethane, and the mixture was heated to 60 °C and stirred overnight. Sodium cyanoborohydride (2.61 g, 41.63 mmol) was added to the reaction mixture, and the reaction was continued at 60 °C for 12 hours. The reaction was quenched by adding saturated ammonium chloride aqueous solution in an ice-water bath. The mixture was extracted three times with dichloromethane, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to give a colorless oily compound ND028-2 (300 mg, 20% yield). 1H NMR (400MHz, DMSO-d6) δ11.41(d,J=2.2Hz,1H),7.70(d,J=8.1Hz,1H),5.70(d,J=1.3Hz, 1H),5.57(dd,J=8.0,2.1Hz,1H),4.39(d,J=5.4Hz,1H),4.30(dd,J=9.0,5.4Hz,1H),4.1 2(dd,J=13.2,2.4Hz,1H),3.93(dd,J=13.3,2.6Hz,1H),3.88(d,J=9.0Hz,1H),2.70(s,2 H), 2.65 (s, 3H), 1.27 (dd, J = 7.4, 4.9Hz, 28H), 1.11–1.03 (m, 28H), 0.89 (t, J = 6.6Hz, 3H). LCMS: m / z=740.1,[M+H] + .

[0721] Step 3. 2'-O-(hexad...

Claims

Compounds of formula (I) or their salts, solvates or isotopically labeled compounds: in: B is H, a base (including modified or unmodified bases), or -L. B -(C 1-30 (hydrocarbon group), of which L B It is a linker; R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support, optionally wherein R1 and R2 are not both C 1-30 hydrocarbon group; X is selected from non-existent, -O-, -S-, -SS-, -OC(=O)-, -C(=O)-O-, -NR4C(=O)-, -C(=O)NR4-, -NR4-, -O-NR4-, -NR4-O-, -O-NR4C(=O)- or -C(=O)NR4-O-; R3 is H or C. 1-30 hydrocarbon group; R B For H or C 1-6 alkyl; Or R B Together with XR3, they form C 2-5 An alkylene chain, wherein the alkylene chain is optionally substituted by one or more groups independently selected from the following: hydroxyl, halogen, cyano, mercapto, azide, nitro, NR. a R b C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 1-6 Alkoxy; R1' represents H and -C 0-6 alkylene-O-(C 1-30 hydrocarbon group), -C 0-6 Alkylene-S-(C 1-30 hydrocarbon group), -C 0-6 Alkylene-NR4-(C 1-30 hydrocarbon group), -C 0-6 Alkylene-C(O)-(C 1-30 (hydrocarbon group) or -C 0-6 Alkylene-C(S)-(C 1-30 (hydrocarbon group); Or R1' is -C 0-6 Alkylene -O-, -C 0-6 Alkylene-S-, -C 0-6 Alkylene-NR4-, -C 0-6 alkylene-C(O)- or -C 0-6 Alkylene-C(S)-,XR3 indicates Furthermore, R1' connects with XR3 to form a bridge ring, where the wavy line indicates that the valence bond is connected to R1', and the asterisk indicates that the valence bond is connected to the rest of the molecule; R m and R n Each is independently H or C 1-6 Alkyl, or R m and R n The connection forms a single bond; R4 is H or C. 1-6 alkyl; R a and R b Each is independently H or C 1-6 alkyl; Wherein, the C 1-30 The hydrocarbon group is optionally selected from hydroxyl, halogen, cyano, mercapto, azide, nitro, NR a R b C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 3-8 Cyclic hydrocarbon group, C 3-8 Cyclic hydrocarbon group, 3-8 membered heterocyclic group, alkynyl group, C 6-10 Aryl or C 5-10 Substitution of heteroaryl groups, and / or optionally, the C 1-30 A C-shaped carbon atom is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 cycloalkyl, wherein formula (I) contains at least one of the C 1-30 Hydrocarbon functional group. The compound according to claim 1, or a salt, solvate, or isotopically labeled compound thereof, wherein formula (I) has the structure of any one of formulas (Ia), (Ib), or (Ic): in, Each variable is defined as described in claim 1. The compound according to claim 1 or 2, or its salt, solvate, or isotopically labeled compound, wherein the C 1-30 The hydrocarbon group is C 12-30 hydrocarbon group, C 14-24 hydrocarbon group, C 16-22 hydrocarbon group or C 16-21 hydrocarbon group; Optionally, the hydrocarbon group is optionally selected from hydroxyl, halogen, cyano, mercapto, azide, nitro, NR. a R b C 1-6 Haloalkyl, C 1- 6-hydroxyalkyl, C 1-6 Alkoxy, C 3-8 Cyclic hydrocarbon group, C 3-8 Cyclic hydrocarbon group, 3-8 membered heterocyclic group, C 6-10 Aryl or C 5-10 Substitution of the heteroaryl group, and / or optionally, the formation of a C12 carbon atom between two adjacent carbon atoms of the hydrocarbon group. 3-6 cycloalkyl, wherein R a and R b Each is independently H or C 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups; Optionally, the hydrocarbon group is optionally selected from hydroxyl, halogen, cyano, mercapto, azide, nitro, NR. a R b C 1-6 Haloalkyl, C 1- 6-hydroxyalkyl, C 1-6 Alkoxy, C 3-8 Cyclic hydrocarbon group, C 3-8 Substitution of the cyclic hydrocarbon group with an oxygen group, and / or optionally, the formation of a C12 carbon atom between two adjacent carbon atoms of the hydrocarbon group. 3-6 cycloalkyl, wherein R a and R b Each is independently H or C 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups; Optionally, the hydrocarbon group is optionally selected from hydroxyl, halogen, cyano, mercapto, azide, nitro, NR. a R b C 1-4 Haloalkyl, C 1- 4-hydroxyalkyl, C 1-4 Alkoxy group substitution, and / or optionally, the formation of a C12 carbon atom between two adjacent carbon atoms of the hydrocarbon group. 3-6 cycloalkyl, wherein R a and R b Each is independently H or C 1-4 Alkyl groups, such as methyl groups; Optionally, the hydrocarbon group is unsubstituted; Optionally, the hydrocarbon group is fully saturated or partially unsaturated, for example, the hydrocarbon group is alkyl, alkenyl or ynyl, preferably fully saturated; Optionally, the hydrocarbon group is straight-chain or branched, preferably straight-chain; Optionally, the hydrocarbon group is connected to the rest of the molecule via a carbon atom at position 1 or a carbon atom at position 3. The compound according to any one of the preceding claims, or its salt, solvate, or isotopically labeled compound, wherein the hydrocarbon group is alkyl, alkenyl, or alkynyl, preferably alkyl; optionally, the hydrocarbon group is C 1-30 Alkyl, such as C 12-30 Alkyl, C 14-24 Alkyl, C 16-22 Alkyl or C 16-21 Alkyl group, optionally the alkyl group is straight-chain or branched; optionally, the hydrocarbon group is C10. 2-30 alkenyl groups, such as C 12-30 alkenyl, C 14- 24 alkenyl, C 16-22 alkenyl or C 16-21 Alkenyl group, optionally the alkenyl group is straight-chain or branched; optionally, the hydrocarbon group is C 2-30 Alkyne groups, such as C 12-30 alkynyl group, C 14-24 alkynyl group, C 16-22 alkynyl or C 16-21 The alkynyl group, optionally the hydrocarbon group is straight-chain or branched; Optionally, the hydrocarbon group is C 16 C 17 C 18 C 19 C 20 C 21 Or C 22 Hydrocarbon groups, such as hexadecyl, hexadecylene, hexadecadienyl, heptadecanyl, heptadecanyl, heptadecanyl, octadecyl, octadecylene, octadecadienyl, octadectrienyl, nonadecanyl, nonadecanylene, nonadecanadienyl, nonadecantrienyl, eicosyl, eicosylene, eicosyladienyl, eicosyltrienyl, dodecyl, dodecylene, dodecylene, dodecylene, 6-octyltetradecyl, 10-hexylhexadecyl, all cis 7,10,13,16,19-docosapentaenyl, all cis 4,7,10,13,16,19-docosahexaenyl, all cis Formula 13,16-docosadienyl, all-cis-7,10,13,16-docosatetraenyl, all-cis-4,7,10,13,16-docosapentenyl or cis-13-docosaenyl, particularly 1-docoalkyl, 6-octyltetradecane-1-yl, 10-hexylhexadecane-1-yl, cis-docosa-13-en-1-yl, docosane-9-yl, docosane-2-yl, docosane-10-yl, docosane-11-yl or cis-4,7,10,13,16,19-docosahexaen-1-yl, wherein the hydrocarbon group may optionally be substituted with a group selected from the following: hydroxyl, amino, cyano, nitro, halogen, C 1-6 Alkoxy, C 3-8 Cyclic hydrocarbon group, C 3-8 Cycloalkyloxy groups, 3-8 membered heterocyclic groups, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, mercapto, azide, alkynyl, aryl, or heteroaryl, or optionally, a C10 group is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 Cycloalkyl; optionally, the hydrocarbon group is 1-hexadecyl, 1-eicosyl, 1-monoalkyl or 1-docoalkyl. The compound or its salt, solvate or isotopically labeled compound according to any one of the preceding claims, wherein B is H. The compound according to any one of claims 1-4, or its salt, solvate, or isotopically labeled compound, wherein, B is a base, such as a modified or unmodified base, such as a natural base (e.g., adenine, guanine, cytosine, thymine, or uracil) or any nucleobase that can pair with a natural base without substantially affecting unwinding behavior, intracellular enzyme recognition, or the activity of the iRNA double strand. Optionally, B is a modified or unmodified purine or pyrimidine base; preferably, B is a modified or unmodified adenine, guanine, xanthine, hypoxanthine, cytosine, thymine, or uracil base; more preferably, B is a modified or unmodified adenine, guanine, cytosine, thymine, or uracil base; optionally, the modification is substitution, denitrification, glycosylation, and / or hydrogenation, and optionally, for substitution modification, the substituent is one or more functional groups or ligands, such as those selected from hydroxyl, halogen, cyano, azide, nitro, thio (=S) (e.g., carbonyl oxygen is substituted by a thio group), NR a N b 、(NR a N b )-C 1-6 Alkylene, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 1-6 Alkyl group, C 1-6 Alkyl-C(=S)-, C 6-10 Aryl, C 6-10 Aryl-C 1-6 Alkylene-, C 6-10 Aryl-C 1-6 imidene-, C 6-10 Aryl-C 1-6 Ethyne-, C 5-10 heteroaryl, C 5-10 heteroaryl-C 1-6 Alkylene-, C 5-10 heteroaryl-C 1-6 imide- and C 5-10 heteroaryl-C 1-6 Imyynyl-, where R a and R b Each is independently H or C 1-6 Alkyl groups, preferably H or C 1-4 Alkyl groups, such as methyl groups; Optionally, B is a modified or unmodified adenine base, guanine base, xanthine base, hypoxanthine base, cytosine base, thymine base, or uracil base, wherein the modification is optionally performed by C. 1-6 Alkyl group, C 1-6 Alkyl-C(=S)-, phenyl-C 1-4 alkylene-, phenyl-C 1-6 alkenyl- or phenyl-C 1-6 Isomeryl-substituted, for example, by C 1-6 Alkyl (such as butyryl) or phenyl-C 1-4 Alkyl-(e.g., benzyl) substitution; Optionally, B is a modified or unmodified adenine, guanine, cytosine, thymine, or uracil base, wherein the modification is optionally performed by C. 1-6 Alkyl group, C 1-6 Alkyl-C(=S)-, phenyl-C 1-4 alkylene-, phenyl-C 1-6 alkenyl- or phenyl-C 1-6 Isomeryl-substituted, for example, by C 1-6 Alkyl (such as butyryl) or phenyl-C 1-4 Alkyl-(e.g., benzyl) substitution; Optionally, B is a modified or unmodified adenine, guanine, cytosine, thymine, or uracil base, wherein the modification is optionally performed by C. 1-6 Alkyl (such as butyryl) or benzyl substitution. The compound according to any one of claims 1-4, or its salt, solvate, or isotopically labeled compound, wherein B is -L B -(C 1-30 Hydrocarbon group), preferably -L B -(C 12-30 (hydrocarbon group), more preferably -L B -(C 14-24 Hydrocarbon group), for example -L B -(C 16-22 (hydrocarbon group) or -L B -(C 20- 22 (Hydrocarbon group); optionally, the hydrocarbon group is fully saturated or partially unsaturated, and / or the hydrocarbon group is straight-chain or branched; Optional, B is -L B -(C 1-30 Alkyl groups, such as -L B -(C 12-30 alkyl), -L B -(C 14-24 alkyl), -L B -(C 16-22 alkyl) or -L B -(C 20-22 Alkyl group), wherein the alkyl group is straight-chain or branched; Optional, B is -L B -(C 2-30 alkenyl), for example -L B -(C 12-30 alkenyl), -L B -(C 14-24 alkenyl), -L B -(C 16-22 alkenyl) or -L B -(C 20-22 (alkenyl), wherein the alkenyl group is straight-chain or branched; Optional, B is -L B -(C 2-30 (alkynyl group), such as -L B -(C 12-30 alkynyl group), -L B -(C 14-24 alkynyl group), -L B -(C 16-22 (alkynyl group) or -L B -(C 20-22 (alkynyl group), wherein the alkynyl group is straight-chain or branched. The compound according to any one of the preceding claims, or its salt, solvate, or isotopically labeled compound, wherein L B This indicates non-existence, -O-, -S-, -C(O)-, -C(S)-, -N(R4)-, -C 1-16 Alkylene -, -(NHCH2CH2) m1 -、-(OCH2CH2) m2 -、-(SCH2CH2) m3 - 4-8 member subheterocyclic groups or any combination of two or more thereof, wherein m1, m2 and m3 are each independently 1, 2, 3, 4, 5 or 6; Optional, L B The following characters represent non-existent elements: -O-, -S-, -C(O)-, -C(S)-, -N(R4)-, and -(NHCH2CH2). m1 -、-(NHCH2CH2) m1 -C(O)-、-(NHCH2CH2) m1 -C(S)-、-(OCH2CH2) m2 -、-(OCH2CH2) m2 -C(O)-、-(OCH2CH2) m2 -C(S)-、-(SCH2CH2) m3 -、-(SCH2CH2) m3 -C(O)-、-(SCH2CH2) m3 -C(S)-, 4-8 membered heterocyclic group, -N(R4)-C(O)-, -N(R4)-C(S)-, -N(R4)-C 1-16 Alkylene -C(O)-, -N(R4)-C 1-16 Alkylene-C(S)-, -N(R4)-C(O)-C 1-16 Alkylene-, -N(R4)-C(S)-C 1-16 Alkylene-,-N(R4)-C(O)-C 1- 16 Alkylene -C(O)-N(R4)-, -N(R4)-C(S)-C 1-16 Alkylene -C(O)-N(R4)-, -N(R4)-C(S)-C 1-16 Alkylene -C(S)-N(R4)-, -N(R4)-C(O)-C 1-16 Alkylene-C(S)-N(R4)-, -(4-8 membered heterocyclic)-C 1-16 Alkylene-(OCH2CH2) m2 -CO-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(SCH2CH2) m2 -CO-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(SCH2CH2) m2 -C(S)-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(OCH2CH2) m2 -C(S)-N(R4)-, where m1, m2 and m3 are each independently 1, 2, 3, 4, 5 or 6; Optional, L B This indicates the absence of -O-, -S-, -C(O)-, -C(S)-, -N(R4)-, 4-8 membered subheterocyclic groups, -N(R4)-C(O)-, -N(R4)-C(S)-, -N(R4)-C(O)-C 1-16 Alkylene -C(O)-N(R4)-, -N(R4)-C(S)-C 1-16 Alkylene -C(O)-N(R4)-, -N(R4)-C(S)-C 1-16 Alkylene -C(S)-N(R4)-, -N(R4)-C(O)-C 1-16 Alkylene-C(S)-N(R4)-, -(4-8 membered heterocyclic)-C 1-16 Alkylene-(OCH2CH2) m2 -CO-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(SCH2CH2) m2 -CO-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(SCH2CH2) m2 -C(S)-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(OCH2CH2) m2 -C(S)-N(R4)-, where m2 is 1, 2, 3, 4, 5 or 6; Optional, L B This represents -O-, 4-8 membered subheterocyclic groups, -N(R4)-C(O)-, -N(R4)-C(S)-, and -N(R4)-C(O)-C. 1-16 Alkylene -C(O)-N(R4)-, -N(R4)-C(S)-C 1-16 Alkylene -C(O)-N(R4)-, -N(R4)-C(S)-C 1-16 Alkylene -C(S)-N(R4)-, -N(R4)-C(O)-C 1-16 Alkylene-C(S)-N(R4)-, -(4-8 membered heterocyclic)-C 1-16 Alkylene-(OCH2CH2) m2 -CO-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(SCH2CH2) m2 -CO-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(SCH2CH2) m2 -C(S)-N(R4)-、-(4-8 membered heterocyclic group)-C 1-16 Alkylene-(OCH2CH2) m2 -C(S)-N(R4)-, where m2 is 1, 2, 3, 4, 5 or 6; Optional, L B This represents -O-, 5-6-membered heteroaryl, -N(H)-C(O)-, -N(H)-C(S)-, -N(H)-C(O)-C 1-16 alkylene-C(O)-N(H)-, -(5-6-membered heteroaryl)-C 1-16 Alkylene-(OCH2CH2) m2 -CO-N(H)-, where m2 is 1, 2, 3, 4, 5 or 6; optionally, the 5-6 quinone heteroaryl contains 1, 2 or 3 nitrogen atoms as heteroatoms, for example, a triazolyl group. The compound according to any one of claims 1-4, or its salt, solvate, or isotopically labeled compound, wherein B is selected from: H, adenine, guanine, cytosine, thymine, uracil, ... (For example ), -O-(C 12-30 Alkyl), -N(H)-C(O)-(C 12-30 alkyl), Optionally, the alkyl group is straight-chain or branched, preferably straight-chain; Optionally, B is selected from: adenine, guanine, cytosine, thymine, uracil, (For example )； Optionally, B is selected from: -O-(C 12-30 Alkyl), -N(H)-C(O)-(C 12-30 alkyl), Optionally, the alkyl group is straight-chain or branched, preferably straight-chain; Optionally, B is selected from: -O-(C 22 Straight-chain alkyl), -N(H)-C(O)-(C 21 (linear alkyl) The compound according to any one of the preceding claims, or its salt, solvate, or isotopically labeled compound, wherein one of R1 and R2 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 One is a hydrocarbon group or a solid support, and the other is a reactive phosphorus group; Optionally, R1 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 Hydrocarbon group or solid support, and R2 is a reactive phosphorus group; Optionally, R1 is a reactive phosphorus group, and R2 is a H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or a C. 1-30 Hydrocarbon-based or solid-phase support; Optionally, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group; Optionally, R1 is a reactive phosphorus group, and R2 is a C2 group. 1-30 hydrocarbon group; Optionally, R1 is a reactive phosphorus group and R2 is a hydroxyl protecting group (such as DMTr); Optionally, R1 and R2 are both reactive phosphorus groups, and the reactive phosphorus groups may be the same or different; Optionally, R1 is a solid support and R2 is a hydroxyl protecting group (such as DMTr); Optionally, the hydroxyl protecting group is selected from: trimethylsilyl (TMS), triethylsilyl (TES), dimethylisopropylsilyl (DMIPS), diethylisopropylsilyl (DEIPS), tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), triisopropylsilyl (TIPS), acetyl (Ac), chloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl (TFA), p-methoxybenzoyl, allyloxycarbonyl (Alloc), 2,2,2-trichloroethoxycarbonyl (Troc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), benzyl (Bn), p-methoxybenzyl (P) MB), allyl, triphenylmethyl (Tr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzylmethyl (PMBM), 4,4'-dimethoxytriphenylmethyl (DMTr), monomethoxytriphenylmethyl (MMT), 9-fluorenylmethoxycarbonyl (Fmoc), o-nitrophenylcarbonyl, p-phenylazophenylcarbonyl, benzoyl, p-chlorobenzoyl and 5'-(α-methyl-2-nitropiperyl)oxycarbonyl (MeNPOC), especially 4,4'-dimethoxytriphenylmethyl (DMTr); Optionally, the reactive phosphorus group is derived from: a phosphate group (e.g., -P(=O)(OH)2), a thiophosphate group (e.g., -P(=S)(OH)2), an O,S-thiophosphate group (e.g., -P(=O)(OH)(SH)), a phosphite group (e.g., -P(OH)2), a phosphoramidite group, an activated phosphate group, an activated thiophosphate group, or an activated phosphite group; Optionally, the reactive phosphorus group is derived from or is a phosphoramidite group, for example... Where R a and R b Each is independently H or C 1-6 Alkyl groups, and R c C is the optional replacement. 1-6 Alkyl groups, optionally substituted with hydroxyl, halogen, cyano, azide, NH2, or nitro groups, preferably R c It is a C that can be substituted with a cyano group. 1-6 Alkyl groups; for example, the reactive phosphorus group is derived from or is derived from (2-cyanoethyl-N,N-diisopropyl)phosphamide. Optionally, the reactive phosphorus group is derived from a phosphate ester group, a thiophosphate ester group, or an O,S-thiophosphate ester group; Optionally, the reactive phosphorus group is derived from either a phosphate ester group or a thiophosphate ester group; Optionally, the C 1-30 The hydrocarbon group is C 12-30 Hydrocarbon group, preferably C 14-24 Hydrocarbon group, more preferably C 16-22 The hydrocarbon group is fully saturated or partially unsaturated, and / or the hydrocarbon group is straight-chain or branched; optionally, the hydrocarbon group is C 1-30 Alkyl, such as C 12-30 Alkyl, C 14-24 Alkyl or C 16- 22 Alkyl group, optionally the alkyl group is straight-chain or branched; optionally, the hydrocarbon group is C10. 2-30 alkenyl groups, such as C 12-30 alkenyl, C 14-24 alkenyl or C 16- 22 Alkenyl group, optionally the alkenyl group is straight-chain or branched; optionally, the hydrocarbon group is C 2-30 Alkyne groups, such as C 12-30 alkynyl group, C 14-24 alkynyl or C 16- 22 The alkynyl group, optionally the hydrocarbon group is straight-chain or branched; Optionally, the solid support is a resin, for example... The compound according to any one of the preceding claims, or its salt, solvate, or isotopically labeled compound, wherein X is selected from non-existent, -O-, -S-, -O-NR4-, -NR4-O-, -O-NR4-C(O)-, and -C(=O)NR4-O-, where R4 is H or C. 1-6 Alkyl groups, preferably H or C 1-4 Alkyl groups such as methyl; preferably, X is selected from absent, -O-, -O-NR4-, -NR4-O-, and -O-NR4-C(O)-, wherein R4 is H or C. 1-6 Alkyl groups, preferably H or C 1-4 Alkyl groups such as methyl; and R3 is H. Or R3 is C 1-6 Hydrocarbon groups, such as C 1-6 Alkyl (such as methyl), C 2-6 alkenyl or C 2-6 alkynyl group; Or R3 is C 1-30 Hydrocarbon group, preferably C 12-30 hydrocarbon group, C 14-24 hydrocarbon group, C 16-22 hydrocarbon group or C 16-21 The hydrocarbon group is fully saturated or partially unsaturated, and / or the hydrocarbon group is straight-chain or branched; optionally, the hydrocarbon group is C 1-30 Alkyl, such as C 12-30 Alkyl, C 14-24 Alkyl, C 16- 22 Alkyl or C 16-21 Alkyl group, optionally the alkyl group is straight-chain or branched; optionally, the hydrocarbon group is C10. 2-30 alkenyl groups, such as C 12-30 alkenyl, C 14- 24 alkenyl, C 16-22 alkenyl or C 16-21 Alkenyl group, optionally the alkenyl group is straight-chain or branched; optionally, the alkynyl group is C 2-30 Alkyne groups, such as C 12- 30 alkynyl group, C 14-24 alkynyl group, C 16-22 alkynyl or C 16-21 The alkynyl group, optionally, is straight-chain or branched; Optionally, R3 is C 16-22 Hydrocarbon groups, such as C 16 C 17 C 18 C 19 C 20 C 21 Or C 22 Hydrocarbon groups, such as hexadecyl, hexadecylene, hexadecadienyl, heptadecanyl, heptadecanyl, heptadecanyl, octadecyl, octadecylene, octadecadienyl, octadectrienyl, nonadecanyl, nonadecanylene, nonadecanadienyl, nonadecantrienyl, eicosyl, eicosylene, eicosyladienyl, eicosyltrienyl, dodecyl, dodecylene, dodecylene, dodecylene, 6-octyltetradecyl, 10-hexylhexadecyl, all cis 7,10,13,16,19-docosapentaenyl, all cis 4,7,10,13,16,19-docosahexaenyl, all cis Formula 13,16-docosadienyl, all-cis-7,10,13,16-docosatetraenyl, all-cis-4,7,10,13,16-docosapentenyl or cis-13-docosaenyl, particularly 1-docoalkyl, 6-octyltetradecane-1-yl, 10-hexylhexadecane-1-yl, cis-docosa-13-en-1-yl, docosane-9-yl, docosane-2-yl, docosane-10-yl, docosane-11-yl or cis-4,7,10,13,16,19-docosahexaen-1-yl, wherein the hydrocarbon group may optionally be substituted with a group selected from the following: hydroxyl, amino, cyano, nitro, halogen, C 1-6 Alkoxy, C 3-8 Cyclic hydrocarbon group, C 3-8 Cycloalkyloxy groups, 3-8 membered heterocyclic groups, C 1-6 Alkylamino, di(C) 1-6 Alkyl)amino, mercapto, azide, alkynyl, aryl, or heteroaryl, or optionally, a C10 group is formed between two adjacent carbon atoms of the hydrocarbon group. 3-6 Cycloalkyl; preferably, R3 is 1-hexadecyl, 1-eicosyl, 1-monoalkyl or 1-docoalkyl. The compound according to any one of the preceding claims, or its salt, solvate, or isotopically labeled compound, wherein R B For H or C 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups, preferably R B For H; Or, R B Together with XR3, they form C 2-5 The alkylene chain, such as a C3 or C4 alkylene chain, is optionally substituted by one or more groups independently selected from the following: hydroxyl, halogen, cyano, mercapto, azide, nitro, NR. a R b C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 1-6 alkoxy group, where R a and R b Each is independently H or C 1-6 Alkyl, preferably C 1-4 Alkyl groups such as methyl; preferably, R B Together with XR3, they form C 2-5 Alkylene chain, such as a C3 or C4 alkylene chain; optionally, (I) has the structure of formula (Id): in, Each variable is as defined in the preceding claims. The compound according to any one of the preceding claims, or its salt, solvate, or isotopically labeled compound, wherein R1' represents H and -C 0-6 alkylene-O-(C 1-30 hydrocarbon group), -C 0-6 Alkylene-S-(C 1-30 hydrocarbon group), -C 0-6 Alkylene-NR4-(C 1-30 hydrocarbon group), -C 0-6 Alkylene-C(O)-(C 1-30 hydrocarbon group), -C 0-6 Alkylene-C(S)-(C 1-30 (Hydrocarbon group); preferably, R1' is H or -C 0-6 alkylene-O-(C 1-30 (hydrocarbon group) or -C 0- 6-alkylene-S-(C 1-30 Hydrocarbon group), preferably H or -C 0-6 alkylene-O-(C 1-30 Hydrocarbon group), more preferably H or -C 1-6 alkylene-O-(C 1-30 (hydrocarbon group); Optionally, the C 1-30 The hydrocarbon group is C 12-30 hydrocarbon group, C 14-24 hydrocarbon group, C 16-22 hydrocarbon group or C 16-21 The hydrocarbon group is fully saturated or partially unsaturated, and / or the hydrocarbon group is straight-chain or branched; optionally, the hydrocarbon group is C 1-30 Alkyl, such as C 12-30 Alkyl, C 14-24 Alkyl, C 16-22 Alkyl or C 16-21 Alkyl group, optionally the alkyl group is straight-chain or branched; optionally, the hydrocarbon group is C10. 2-30 alkenyl groups, such as C 12- 30 alkenyl, C 14-24 alkenyl, C 16-22 alkenyl or C 16-21 Alkenyl group, optionally the alkenyl group is straight-chain or branched; optionally, the alkynyl group is C 2- 30 Alkyne groups, such as C 12-30 alkynyl group, C 14-24 alkynyl group, C 16-22 alkynyl or C 16-21 The alkynyl group, optionally, is straight-chain or branched. The compound according to any one of the preceding claims, or its salt, solvate, or isotopically labeled compound, wherein R1' is -C 0-6 Alkylene -O-, -C 0-6 Alkylene-S-, -C 0-6 Alkylene-NR4-, -C 0-6 Alkylene -C(O)-, -C 0-6 Alkylene -C(S)-, preferably -O-, -S-, -NR4-, -C(O)-, -C(S)-, more preferably -C(O)- or -C(S)-, XR3 indicates Furthermore, R1' connects with XR3 to form a bridge ring, where the wavy line indicates that the valence bond is connected to R1', and the asterisk indicates that the valence bond is connected to the rest of the molecule; Optionally, R1' is -C(O)-, and XR3 represents... Furthermore, R1' connects with XR3 to form a bridge ring, where the wavy line indicates that the valence bond is connected to R1', and the asterisk indicates that the valence bond is connected to the rest of the molecule; Optionally, equation (I) has the structure shown in equation (Ie): The compound according to any one of claims 1-14, or a salt, solvate, or isotopically labeled compound thereof, wherein formula (I) has the structure of formula (Ia): in: B is H, a base (including modified or unmodified bases), or -L. B -(C 1-30 (hydrocarbon group), of which L B It is a linker; preferably, B is H or a base (including modified or unmodified bases); R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 hydrocarbon group; X is selected from non-existent, -O-, -O-NR4-, -NR4-O-, and -O-NR4-C(O)-; R3 is H or C. 1-30 Hydrocarbon group, preferably C 1-30 hydrocarbon group; R B For H or C 1-6 Alkyl groups, preferably H; R1' is H or -C 0-6 alkylene-O-(C 1-30 Hydrocarbon group), preferably H; R4 is H or C. 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups; Wherein C 1-30 The hydrocarbon group is fully saturated or partially unsaturated, and wherein formula (Ia) contains at least one of the C groups. 1-30 Hydrocarbon functional group; optionally, the C 1-30 The hydrocarbon group is C 12-30 Hydrocarbon group, preferably C 14-24 Hydrocarbon group, more preferably C 16-22 hydrocarbon group; Optionally, the reactive phosphorus group is derived from a phosphate ester group, a thiophosphate ester group, an O,S-thiophosphate ester group, or a phosphoramidite group. Optionally, R1 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 Hydrocarbon group or solid support, and R2 is a reactive phosphorus group; Optionally, R1 is a reactive phosphorus group, and R2 is a H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or a C. 1-30 Hydrocarbon-based or solid-phase support; Optionally, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group; Optionally, R1 is a reactive phosphorus group, and R2 is a C2 group. 1-30 hydrocarbon group; Optionally, R1 is a reactive phosphorus group and R2 is a hydroxyl protecting group (such as DMTr); Optionally, R1 and R2 are both reactive phosphorus groups, and the reactive phosphorus groups may be the same or different; Optionally, R1 is a solid support and R2 is a hydroxyl protecting group (such as DMTr). The compound according to claim 15, or its salt, solvate, or isotopically labeled compound, wherein... B is H; R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 hydrocarbon group; X is non-existent, -O-, -O-NR4-, -NR4-O-, or -O-NR4-C(O)-; preferably, it is non-existent, -O-, -O-NR4-, or -NR4-O-; R3 is H or C. 1-30 Hydrocarbon group, preferably C 1-30 hydrocarbon group; R B For H or C 1-6 Alkyl groups, preferably H; R1' is H or -C 0-6 alkylene-O-(C 1-30 Hydrocarbon group), preferably H; R4 is H or C. 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups; Wherein C 1-30 The hydrocarbon group is fully saturated or partially unsaturated, and wherein formula (Ia) contains at least one of the C groups. 1-30 hydrocarbon functional groups; Or, among them: B is a base (including modified or unmodified bases); R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 Hydrocarbon group; preferably, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group; X is selected from -O-, -O-NR4-, -NR4-O-, and -O-NR4-C(O)-; R3 is H or C. 1-30 Hydrocarbon group, preferably C 1-30 hydrocarbon group; R B For H or C 1-6 Alkyl groups, preferably H; R1' is H or -C 0-6 alkylene-O-(C 1-30 Hydrocarbon group), preferably H; R4 is H or C. 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups; Wherein C 1-30 The hydrocarbon group is fully saturated or partially unsaturated, and wherein formula (Ia) contains at least one of the C groups. 1-30 hydrocarbon functional groups; Or one of them B is H or a base (including modified or unmodified bases), preferably a base (including modified or unmodified bases); R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 Hydrocarbon group; preferably, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group; X is selected from -O-; R3 is C 1-6 A hydrocarbon group, wherein the hydrocarbon group is fully saturated or partially unsaturated; R B For H or C 1-6 Alkyl groups, preferably H; R1' is -C 0-6 alkylene-O-(C 1-30 Hydrocarbon group), preferably -C 1-6 alkylene-O-(C 1-30 (hydrocarbon group); Wherein C 1-30 The hydrocarbon group is either fully saturated or partially unsaturated. The compound according to any one of claims 1-14, or a salt, solvate, or isotopically labeled compound thereof, wherein formula (I) has the structure of formula (Ib): in: B is H, a base (including modified or unmodified bases), or -L. B -(C 1-30 (hydrocarbon group), of which L B It is a linker; preferably, B is H or a modified or unmodified base; R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 hydrocarbon group; X is selected from non-existent, -O-, -O-NR4-, -NR4-O-, and -O-NR4-C(O)-; R3 is H or C. 1-30 Hydrocarbon group, preferably C 1-30 hydrocarbon group; R4 is H or C. 1-6 Alkyl, such as C 1-4 Alkyl groups, such as methyl groups; Wherein C 1-30 The hydrocarbon group is fully saturated or partially unsaturated, and wherein formula (Ib) contains at least one of the C groups. 1-30 Hydrocarbon functional group. Optionally, the C 1-30 The hydrocarbon group is C 12-30 Hydrocarbon group, preferably C 14-24 Hydrocarbon group, more preferably C 16-22 hydrocarbon group; Optionally, the reactive phosphorus group is derived from a phosphate ester group, a thiophosphate ester group, an O,S-thiophosphate ester group, or a phosphoramidite group. Optionally, R1 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 Hydrocarbon group or solid support, and R2 is a reactive phosphorus group; Optionally, R1 is a reactive phosphorus group, and R2 is a H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or a C. 1-30 Hydrocarbon-based or solid-phase support; Optionally, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group; Optionally, R1 is a reactive phosphorus group, and R2 is a C2 group. 1-30 hydrocarbon group; Optionally, R1 is a reactive phosphorus group and R2 is a hydroxyl protecting group (such as DMTr); Optionally, R1 and R2 are both reactive phosphorus groups, and the reactive phosphorus groups may be the same or different; Optionally, R1 is a solid support and R2 is a hydroxyl protecting group (such as DMTr). The compound according to any one of claims 1-14, or a salt, solvate, or isotopically labeled compound thereof, wherein formula (I) has the structure of formula (Ic): in: B is H or a base (including modified or unmodified bases), preferably a base (including modified or unmodified bases); R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 hydrocarbon group; X is selected from non-existent, -O-, -O-NR4-, -NR4-O-, and -O-NR4-C(O)-, for example, -NR4-O-; R3 is H or C. 1-30 Hydrocarbon group, preferably C 1-30 hydrocarbon group; R B For H or C 1-6 Alkyl groups, preferably H; R1' is H; Wherein C 1-30 The hydrocarbon group is fully saturated or partially unsaturated, and wherein formula (Ic) contains at least one of the C groups. 1-30 Hydrocarbon functional group; optionally, the C 1-30 The hydrocarbon group is C 12-30 Hydrocarbon group, preferably C 14-24 Hydrocarbon group, more preferably C 16-22 hydrocarbon group; Optionally, the reactive phosphorus group is derived from a phosphate ester group, a thiophosphate ester group, an O,S-thiophosphate ester group, or a phosphoramidite group. Optionally, R1 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 Hydrocarbon group or solid support, and R2 is a reactive phosphorus group; Optionally, R1 is a reactive phosphorus group, and R2 is a H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or a C. 1-30 Hydrocarbon-based or solid-phase support; Optionally, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group; Optionally, R1 is a reactive phosphorus group, and R2 is a C2 group. 1-30 hydrocarbon group; Optionally, R1 is a reactive phosphorus group and R2 is a hydroxyl protecting group (such as DMTr); Optionally, R1 and R2 are both reactive phosphorus groups, and the reactive phosphorus groups may be the same or different; Optionally, R1 is a solid support and R2 is a hydroxyl protecting group (such as DMTr). The compound according to any one of claims 1-14, or a salt, solvate, or isotopically labeled compound thereof, wherein formula (I) has the structure of formula (Id): in: B is -L B -(C 1-30 (hydrocarbon group), of which L B It is a linker; R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 hydrocarbon group; R1' is H or -C 0-6 alkylene-O-(C 1-30 Hydrocarbon group), preferably H; Optionally, the C 1-30 The hydrocarbon group is fully saturated or partially unsaturated, and the formula (Id) contains at least one of the C groups. 1-30 Hydrocarbon functional group; optionally, the C 1-30 The hydrocarbon group is C 12-30 Hydrocarbon group, preferably C 14-24 Hydrocarbon group, more preferably C 16-22 hydrocarbon group; Optional, L B This represents -O-, 5-6-membered heteroaryl, -N(H)-C(O)-, -N(H)-C(S)-, -N(H)-C(O)-C 1-16 alkylene-C(O)-N(H)-, -(5-6-membered heteroaryl)-C 1-16 Alkylene-(OCH2CH2) m2 -CO-N(H)-, where m2 is 1, 2, 3, 4, 5 or 6; preferably, L B This represents -O-, 5-6-membered heteroaryl, -N(H)-C(O)-, -N(H)-C(S)-, -N(H)-C(O)-C 1-12 alkylene-C(O)-N(H)-, -(5-6-membered heteroaryl)-C 1-6 Alkylene-(OCH2CH2) m2 -CO-N(H)-, where m2 is 1, 2, 3, 4, 5 or 6; optionally, the 5-6-membered heteroaryl contains 1, 2 or 3 nitrogen atoms as heteroatoms, for example, a triazolyl group; Optionally, the reactive phosphorus group is derived from a phosphate ester group, a thiophosphate ester group, an O,S-thiophosphate ester group, or a phosphoramidite group. Optionally, the solid support is a resin, for example... Optionally, R1 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 Hydrocarbon group or solid support, and R2 is a reactive phosphorus group; Optionally, R1 is a reactive phosphorus group, and R2 is a H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or a C. 1-30 Hydrocarbon-based or solid-phase support; Optionally, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group; Optionally, R1 is a reactive phosphorus group, and R2 is a C2 group. 1-30 hydrocarbon group; Optionally, R1 is a reactive phosphorus group and R2 is a hydroxyl protecting group (such as DMTr); Optionally, R1 and R2 are both reactive phosphorus groups, and the reactive phosphorus groups may be the same or different; Optionally, R1 is a solid support and R2 is a hydroxyl protecting group (such as DMTr). The compound according to any one of claims 1-14, or a salt, solvate, or isotopically labeled compound thereof, wherein formula (I) has formula (Ie): in: B is a base (including modified or unmodified bases); R1 and R2 are independently H, hydroxyl protecting group, reactive phosphorus group, and C. 1-30 hydrocarbon group, C 1-30 Hydrocarbon-based or solid support; optionally, R1 and R2 are not both C 1-30 hydrocarbon group; R3 is C 1-30 hydrocarbon group; R B For H or C 1-6 Alkyl groups, preferably H; Wherein C 1-30 The hydrocarbon group is either fully saturated or partially unsaturated; Optionally, the reactive phosphorus group is derived from a phosphate ester group, a thiophosphate ester group, an O,S-thiophosphate ester group, or a phosphoramidite group. Optionally, R1 is H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or C. 1-30 Hydrocarbon group or solid support, and R2 is a reactive phosphorus group; Optionally, R1 is a reactive phosphorus group, and R2 is a H, a hydroxyl protecting group (such as DMTr), a reactive phosphorus group, or a C. 1-30 Hydrocarbon-based or solid-phase support; Optionally, R1 is a hydroxyl protecting group (such as DMTr), and R2 is a reactive phosphorus group; Optionally, R1 is a reactive phosphorus group, and R2 is a C2 group. 1-30 hydrocarbon group; Optionally, R1 is a reactive phosphorus group and R2 is a hydroxyl protecting group (such as DMTr); Optionally, R1 and R2 are both reactive phosphorus groups, and the reactive phosphorus groups may be the same or different; Optionally, R1 is a solid support and R2 is a hydroxyl protecting group (such as DMTr). The compound according to any one of claims 1-14, or a salt, solvate, or isotopically labeled compound thereof, wherein formula (I) has the structure of formula (If) or (If'), Wherein B, X and R3 are as defined in any one of claims 1-14; Optionally, B is H or a base (including modified or unmodified bases); Optionally, X is selected from -O-, *-O-NR4-, *-NR4-O- and *-O-NR4-CO-, and preferably from -O-, *-ON(CH3)-, *-N(CH3)-O-, *-O-NH-CO- and *-ON(CH3)-CO-, wherein the valence bond indicated by * is connected to the pentose ring; Optionally, R3 is C 1-30 hydrocarbon group, Optionally, the formula (I) has the structure of formulas (If-1), (If-2), (If-3), and (If-4). Wherein B is defined as in any one of claims 1-14, preferably H or a base (including modified or unmodified bases). The compound according to claim 21, or a salt, solvate, or isotopically labeled compound thereof, wherein formula (I) has the following structure, Wherein X and R3 are as defined in claim 21, for example as defined in formula (If). The compound according to claim 19, or a salt, solvate, or isotopically labeled compound thereof, wherein formula (I) has the structure of formula (Ig) or (Ig'). in, B is as defined in claim 19. According to claim 1, the compound or its salt, solvate or isotopically labeled compound, wherein the compound of formula (I) is selected from: A dsRNA activator or a pharmaceutically acceptable salt thereof The dsRNA activator comprises a sense strand and an antisense strand capable of forming a double-stranded region. The antisense strand comprises a complementary region complementary to at least 15, 16, 17, 18, 19, 20, 21, 22, or 23 consecutive nucleotides in the target sequence, and the complementary region contains no more than 3, 2, or 1 nucleotide mismatch with the at least 15 consecutive nucleotides in the target sequence. The target sequence is derived from any 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 consecutive nucleotides, for example, 19-23 consecutive nucleotides, of the mRNA of the target gene. The skeletal structure of a compound as defined in any one of claims 1-24, or a salt, solvate, or isotopically labeled compound thereof, is added to the 5' end and / or 3' end of the positive and / or negative strands, or is replaced at one or more internal locations of the positive and / or negative strands with the skeletal structure of a compound as defined in any one of claims 1-24, or a salt, solvate, or isotopically labeled compound thereof, wherein... The skeletal structure of the compound of formula (I) or its salt, solvate or isotopically labeled compound is shown in the following formula: Each variable is defined as described in any one of claims 1-24. Wherein, when the corresponding structure is used to replace nucleotides at one or more internal positions of the sense strand and / or antisense strand, Each nucleotide is attached to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide, respectively; preferably, Linked to the 3' carbon of the previous nucleotide or the corresponding position. Linked to the 5' carbon of the next nucleotide or the corresponding position; or When the corresponding structure is added to the 5' or 3' end of the justice chain and / or antisense chain, One of them is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is added to the 5' end of the sense and / or antisense strands, Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 5' carbon of the 5' terminal nucleotide or at the corresponding position; or when the corresponding structure is added to the 3' end of the sense strand and / or antisense strand. Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position; Optionally, the compound of formula (I) has the structure of formula (Ia), and its skeletal structure is shown in the following formula: Optionally, the compound of formula (I) has the structure of formula (Ib), and its skeletal structure is shown in the following formula: Optionally, the compound of formula (I) has the structure of formula (Ic), and its skeletal structure is shown in the following formula: Optionally, the compound of formula (I) has the structure of formula (Id), and its skeletal structure is shown in the following formula: Optionally, the compound of formula (I) has the structure of formula (Ie), and its skeletal structure is shown in the following formula: The dsRNA activator of claim 25 or a pharmaceutically acceptable salt thereof, wherein the skeletal structure of the compound of formula (I) or its salt is as shown in the following formula: Each variable is defined as described in any one of claims 1-24. Wherein, when the corresponding structure is used to replace nucleotides at one or more internal positions of the sense strand and / or antisense strand, Each nucleotide is attached to the 3' carbon or corresponding position of the previous nucleotide, and the other is attached to the 5' carbon or corresponding position of the next nucleotide; preferably, Linked to the 3' carbon of the previous nucleotide or the corresponding position. Linked to the 5' carbon or corresponding position of the next nucleotide; or when the corresponding structure is added to the 5' or 3' end of the sense and / or antisense strands. One of them is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is added to the 5' end of the sense and / or antisense strands, Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 5' carbon of the 5' terminal nucleotide or at the corresponding position; or when the corresponding structure is added to the 3' end of the sense strand and / or antisense strand. Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position. The dsRNA active agent of claim 25 or 26 or a pharmaceutically acceptable salt thereof, wherein, The backbone structure of a compound of formula (I) as defined in any one of claims 1-24 or a salt thereof is added to the 5' end or 3' end of the positive strand, or a nucleotide at one or more internal positions of the positive strand is replaced with the backbone structure of a compound of formula (I) as defined in any one of claims 1-24 or a salt thereof, wherein the internal position is not a position within the cleavage site region of the positive strand, preferably not the 9th-12th or 11th-13th positions counting from the 5' end of the positive strand; more preferably, the internal position is the 6th position counting from the 5' end of the positive strand. The dsRNA activator according to any one of claims 25-27, or a pharmaceutically acceptable salt thereof, in, When the skeletal structure of the compound or its salt is used to replace nucleotides at one or more internal positions of the sense and / or antisense strands, the B in the compound is preferably H, or a base complementary to the base of the nucleotide at the corresponding position of the other strand (modified or unmodified), or -L. B -(C 1-30 (Hydrocarbon group); for example, the compound is selected from ND001, ND002, ND003, ND009, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058 or ND060; preferably, the compound is selected from ND001, ND002, ND009, ND011, ND032 or ND033; When the skeletal structure of the compound or its salt is used to add to the 5' or 3' end of the positive chain, the B in the compound is a base (modified or unmodified), H, or -LB-(C1-30 hydrocarbon group), preferably H or -LB-(C1-30 hydrocarbon group); for example, the compound is selected from ND001, ND002, ND003, ND009, ND010, ND011, ND028, ND029, ND030, ND031, ND0 32, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, ND060, preferably selected from ND009, ND010, ND030, ND031, ND032, ND033, ND036, ND045, ND046, ND047, and ND048; The compounds represented by the compound codes mentioned above are as follows: The dsRNA active agent of any one of claims 25-28 or a pharmaceutically acceptable salt thereof, wherein the sense strand and / or antisense strand comprises one or more structures independently selected from the following, or the skeletal structures of the compounds ND001, ND002, ND003, ND009, ND010, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058 and ND060 are respectively shown in the following structures, wherein the ND numbers of the compounds are the same as those of their skeletal structures: in, Y is oxygen or sulfur, and Optionally, the sense and / or antisense chains comprise one or more structures independently selected from the following, or the skeletal structures of compounds ND001, ND002, ND003, ND009, ND010, ND011, ND028, ND029, ND030, ND031, ND032, ND033, ND0036, ND042, ND0045, ND0046, ND0047, ND0048, ND052, ND053, ND054, ND055, ND056, ND057, ND058, and ND060 are respectively shown in the following structures, wherein the compounds have the same ND number as their skeletal structures: Wherein, when the corresponding structure is used to replace nucleotides at one or more internal positions of the sense strand and / or antisense strand, Each nucleotide is attached to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide, respectively; preferably, Linked to the 3' carbon of the previous nucleotide or the corresponding position. Linked to the 5' carbon of the next nucleotide or the corresponding position; or When the corresponding structure is added to the 5' or 3' end of the justice chain and / or antisense chain, One of them is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other indicates that it is attached to a hydrogen, terminal modification or terminal protecting group; Preferably, when the corresponding structure is added to the 5' end of the sense chain and / or antisense chain, Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 5' carbon of the 5' terminal nucleotide or at the corresponding position; or when the corresponding structure is added to the 3' end of the sense strand and / or antisense strand. Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position. The dsRNA activator of any one of claims 25 to 29 or a pharmaceutically acceptable salt thereof, wherein the antisense strand comprises a complementary region substantially or completely complementary to the target sequence of the target gene, and the double-stranded region formed by the sense and antisense strands is completely or substantially complementary. Optionally, the antisense strand has the same number of nucleotides as the target sequence and is mismatched with the mRNA target sequence at the first nucleotide at the 5' end, for example, being completely complementary in the entire nucleotide sequence region except for the first nucleotide at the 5' end; optionally, the first nucleotide at the 5' end of the antisense strand is A or U, for example, U; Optionally, the bichain region formed by the justice chain and the antisense chain is completely complementary or may contain 1, 2 or 3 mismatches; Optionally, the length of the fully complementary double-stranded region is between 15 and 25 nucleotide pairs, 16 and 24 nucleotide pairs, 17 and 23 nucleotide pairs, or 18 and 22 nucleotide pairs. For example, the length of the fully complementary double-stranded region is 15, 16, 17, 18, or 19 nucleotides, such as 18 or 19 nucleotides. Alternatively, the number of fully complementary nucleotide pairs in the double-stranded region is between 15 and 25 nucleotide pairs, 16 and 25 nucleotide pairs, 16 and 24 nucleotide pairs, 17 and 24 nucleotide pairs, 17 and 23 nucleotide pairs, 18 and 23 nucleotide pairs, or 19 and 22 nucleotide pairs. For example, the number of fully complementary nucleotide pairs in the double-stranded region is 16, 17, 18, 19, 20, or 21 pairs of nucleotides, such as 17, 18, or 19 pairs of nucleotides. Optionally, the lengths of the positive and negative strands are each independently 15-30 nucleotides, for example 17-27 nucleotides, for example 19-25 nucleotides, for example 19-23 nucleotides, or for example 19-21 nucleotides. For example, the length of the positive strand is 19 nucleotides and the length of the negative strand is 21 nucleotides; or the length of the positive strand is 21 nucleotides and the length of the negative strand is 21 nucleotides; or the length of the positive strand is 21 nucleotides and the length of the negative strand is 23 nucleotides. The dsRNA activator of any one of claims 25 to 30 or a pharmaceutically acceptable salt thereof, wherein the dsRNA activator further comprises other modified nucleosides, such as: 2'-O-methyl modified nucleosides, nucleosides containing a 5'-thiophosphate group, terminal nucleosides linked to a cholesterol derivative or a dodecanoic acid didecanoic acid group, locked nucleosides, baseless nucleosides, 2'-deoxyribonucleosides, 2'-fluorinated nucleosides, 2'-amino-modified nucleosides, 2'-alkyl-modified nucleosides, morpholino nucleosides, non-locked nucleosides (UNA), aminophosphates or nucleosides containing non-natural bases, or any combination thereof; Optionally, the oligonucleotide in the dsRNA activator may comprise at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 or more modified nucleosides, or all nucleosides of the oligonucleotide may be modified nucleosides, wherein, For each of the multiple modified nucleosides in the dsRNA activator, the modification is independent and does not need to be the same; Optionally, the dsRNA activator may also optionally contain chemical modifications at the 5' and / or 3' ends, such as phosphate ester modifications at the 5' end, such as 5'-(E)-vinylphosphonate (5'-(E)-VP), 5'-methylphosphonate (5'-MP), (S)-5'-C-methyl analog and 5'-thiophosphate (5'-PS); Optionally, the antisense strand of the dsRNA activator contains 2'-methoxy (2'-O-methyl) modified nucleotides, for example, all nucleotides are 2'-methoxy modified nucleotides or 1-21 nucleotides are 2'-methoxy modified nucleotides, for example, 17 nucleotides are 2'-methoxy modified nucleotides, for example, the nucleotides at positions 1, 3-5, 7-13, 15 and 17-21 of the antisense strand of the dsRNA activator are 2'-methoxy modified nucleotides; Optionally, the positive strand of the dsRNA activator contains 2'-methoxy (2'-O-methyl) modified nucleotides, for example, all nucleotides are 2'-methoxy modified nucleotides, for example, 15, 16 or 17 nucleotides are 2'-methoxy modified nucleotides; Optionally, the dsRNA activator comprises 2'-fluorine modified nucleotides, for example, 1 to 5 nucleotides in the sense strand and / or antisense strand are 2'-fluorine modified nucleotides, for example, 3, 4 or 5 nucleotides in the sense strand and / or antisense strand are 2'-fluorine modified nucleotides. The dsRNA active agent of any one of claims 25 to 31, or a pharmaceutically acceptable salt thereof, comprises a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand and the antisense strand each have the following modification patterns: in, Nf = any 2'-fluorine modified nucleotide Nm = any 2'-methoxynucleotide; s represents a thiophosphate bond; VP represents a 5'-vinyl phosphonate (5'-VP) modification at the 5' end of the antisense strand, such as 5'-(E)-VP modification; (ND) = The skeletal structure of a compound as defined in any one of claims 1-24 or a salt, solvate or isotopically labeled compound thereof, such as that defined in any one of claims 25-29; For example, the 5' nucleotide VPNms of the antisense strand can be VPUms, with the following structure: The dsRNA activator of claim 32 or a pharmaceutically acceptable salt thereof, comprising a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand and the antisense strand each have the following modification patterns: in, Nf = any nucleotide modified with 2'-fluorine; Nm = any 2'-methoxynucleotide; 's' represents a thiophosphate bond; VP indicates that the 5' end nucleotide of the antisense strand has a 5'-vinyl phosphonate (5'-VP) modification, such as 5'-(E)-VP modification; For example, the 5' nucleotide VPNms of the antisense strand can be VPUms, with the following structure: The structures of ND001', ND002', ND009', ND010'(ND010'-2), ND011', ND031', ND032', ND033', ND036', ND046' and ND048' are as defined in claim 29. The dsRNA activator of claim 33 or a pharmaceutically acceptable salt thereof, comprising a sense strand and an antisense strand forming a double-stranded region, wherein the dsRNA is used to inhibit SOD1 and comprises a sense strand and an antisense strand, wherein the antisense strand and the sense strand each comprise or consist of the nucleotide sequence shown in SEQ ID NO: Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:10; Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:11; Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:12; Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:13; Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:14; Antisense chain: SEQ ID NO:9; Justice chain: SEQ ID NO:15; Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:16; Antisense chain: SEQ ID NO:9; Justice chain: SEQ ID NO:17; Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:18; Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:19; Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:20; or Antisense chain: SEQ ID NO:8; Justice chain: SEQ ID NO:21; or, The dsRNA is used to inhibit ACVR1C and comprises a sense strand and an antisense strand, wherein the antisense strand and the sense strand each comprise or consist of the nucleotide sequence shown in SEQ ID NO:: Antisense chain: SEQ ID NO:22; Justice chain: SEQ ID NO:25; Antisense chain: SEQ ID NO:22; Justice chain: SEQ ID NO:26; Antisense chain: SEQ ID NO:23; Justice chain: SEQ ID NO:27; or Antisense chain: SEQ ID NO:23; Justice chain: SEQ ID NO:

28. An oligonucleotide chain or a pharmaceutically acceptable salt thereof, comprising at one or more positions the skeletal structure of a compound of formula (I) as defined in any one of claims 1-24 or a salt thereof, the skeletal structure of said compound of formula (I) or a salt thereof being as follows: Each variable is defined as described in any one of claims 1-24. Wherein, when the corresponding structure is located at one or more internal positions of the oligonucleotide chain, Each nucleotide is attached to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide, respectively; preferably, Linked to the 3' carbon of the previous nucleotide or the corresponding position. Linked to the 5' carbon of the next nucleotide or the corresponding position; or When the corresponding structure is located at the 5' end or 3' end of the oligonucleotide chain, One of them is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is located at the 5' end of the oligonucleotide chain, Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 5' carbon of the 5' terminal nucleotide or at the corresponding position; or when the corresponding structure is located at the 3' end of the oligonucleotide chain. Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position; Optionally, the compound of formula (I) has the structure of formula (Ia), and its skeletal structure is shown in the following formula: Optionally, the compound of formula (I) has the structure of formula (Ib), and its skeletal structure is shown in the following formula: Optionally, the compound of formula (I) has the structure of formula (Ic), and its skeletal structure is shown in the following formula: Optionally, the compound of formula (I) has the structure of formula (Id), and its skeletal structure is shown in the following formula: Optionally, the compound of formula (I) has the structure of formula (Ie), and its skeletal structure is shown in the following formula: Optionally, the compound is located at the 5' end, 3' end, or internal position of the oligonucleotide chain, for example, at the 6th position counting from the 5' end. The oligonucleotide chain of claim 35 or a pharmaceutically acceptable salt thereof, wherein the skeletal structure of the compound of formula (I) or its salt is as shown in the following formula: Each variable is defined as described in any one of claims 1-24. Wherein, when the corresponding structure is located at one or more internal positions of the oligonucleotide chain, Each nucleotide is attached to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide, respectively; preferably, Linked to the 3' carbon of the previous nucleotide or the corresponding position. Linked to the 5' carbon of the next nucleotide or the corresponding position; or When the corresponding structure is located at the 5' end or 3' end of the oligonucleotide chain, One of them is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is located at the 5' end of the oligonucleotide chain, Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 5' carbon of the 5' terminal nucleotide or at the corresponding position; or when the corresponding structure is located at the 3' end of the oligonucleotide chain. Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position. The oligonucleotide chain of claim 35 or 36, or a pharmaceutically acceptable salt thereof, comprising one or more structures independently selected from the following: in, Y is oxygen or sulfur, and Optionally, the justice chain and / or antisense chain includes one or more structures independently selected from the following: Wherein, when the corresponding structure is located at one or more internal positions of the oligonucleotide chain, Each nucleotide is attached to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide, respectively; preferably, Linked to the 3' carbon of the previous nucleotide or the corresponding position. Linked to the 5' carbon of the next nucleotide or the corresponding position; or When the corresponding structure is located at the 5' end or 3' end of the oligonucleotide chain, One of them is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is located at the 5' end of the oligonucleotide chain, Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 5' carbon of the 5' terminal nucleotide or at the corresponding position; or when the corresponding structure is located at the 3' end of the oligonucleotide chain. Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position. A pharmaceutical composition comprising a dsRNA active agent as defined in any one of claims 25-34 or a pharmaceutically acceptable salt thereof, or an oligonucleotide chain as defined in any one of claims 35-37 or a pharmaceutically acceptable salt thereof. The use of the dsRNA activator as defined in any one of claims 25-34 or a pharmaceutically acceptable salt thereof, or the oligonucleotide chain as defined in any one of claims 35-37 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 38, in the manufacture or preparation of a medicament for diseases and / or conditions related to a gene (e.g., SOD1 or ACVR1C gene) targeted by the dsRNA activator, such as diseases and / or conditions caused by abnormal expression and / or activity of a gene (e.g., SOD1 or ACVR1C gene) targeted by the dsRNA activator, or diseases and / or conditions for which one can benefit from inhibiting the expression and / or activity of a gene (e.g., SOD1 or ACVR1C gene) targeted by the dsRNA activator, such as skeletal muscle disorders, cardiomyopathy, or adipose tissue disorders, such as SOD1 or ACVR1C-related diseases or conditions. A method for preventing or treating a disease in a subject, comprising administering to the subject an effective amount of the dsRNA active agent as defined in any one of claims 25-34 or a pharmaceutically acceptable salt thereof, or an oligonucleotide chain as defined in any one of claims 35-37 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 38. Optionally, the disease is a disease and / or condition related to a gene (e.g., SOD1 or ACVR1C gene) targeted by the dsRNA activator, such as a disease and / or condition caused by abnormal expression and / or activity of a gene (e.g., SOD1 or ACVR1C gene) targeted by the dsRNA activator, or a disease and / or condition that can benefit from inhibiting the expression and / or activity of a gene (e.g., SOD1 or ACVR1C gene) targeted by the dsRNA activator, such as skeletal muscle disease, cardiomyopathy, or adipose tissue disease, such as SOD1 or ACVR1C-related diseases or conditions. The use of a compound of formula (I) as defined in any one of claims 1 to 24, or a salt thereof, for the preparation of a dsRNA active agent that can be effectively delivered to a specific tissue or cell, such as muscle tissue (e.g., skeletal muscle tissue or cardiac tissue) or cell, or adipose tissue or cell. A method for preparing a dsRNA activator for delivery to a specific tissue or cell, such as muscle tissue (e.g., skeletal muscle tissue or cardiac muscle tissue) or cell, or adipose tissue or cell, the method comprising adding a backbone structure of a compound as defined in any one of claims 1-24 or a salt thereof, solvate thereof, or isotopically labeled compound to the 5' end and / or 3' end of a double-stranded RNA and / or replacing nucleosides at one or more positions of the double-stranded RNA with a backbone structure of a compound as defined in any one of claims 1-24 or a salt thereof, solvate thereof, or isotopically labeled compound, such that the backbone structure of a compound of formula (I) or a salt thereof is contained at one or more positions of the double-stranded RNA, wherein, The skeletal structure of the compound of formula (I) or its salt is shown in the following formula: Each variable is defined as described in any one of claims 1-24. Wherein, when the corresponding structure is located at one or more internal positions of the oligonucleotide chain, Each nucleotide is attached to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide, respectively; preferably, Linked to the 3' carbon of the previous nucleotide or the corresponding position. Linked to the 5' carbon of the next nucleotide or the corresponding position; or When the corresponding structure is located at the 5' end or 3' end of the oligonucleotide chain, One of them is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is located at the 5' end of the oligonucleotide chain, Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 5' carbon of the 5' terminal nucleotide or at the corresponding position; or when the corresponding structure is located at the 3' end of the oligonucleotide chain. Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position; Optionally, the compound of formula (I) has the structure of formula (Ia), and its skeletal structure is shown in the following formula: Optionally, the compound of formula (I) has the structure of formula (Ib), and its skeletal structure is shown in the following formula: Optionally, the compound of formula (I) has the structure of formula (Ic), and its skeletal structure is shown in the following formula: Optionally, the compound of formula (I) has the structure of formula (Id), and its skeletal structure is shown in the following formula: Optionally, the compound of formula (I) has the structure of formula (Ie), and its skeletal structure is shown in the following formula: The method of claim 42, wherein the skeletal structure of the compound of formula (I) is as shown in formula (I”): Each variable is defined as described in any one of claims 1-24. Wherein, when the corresponding structure is used to replace nucleotides at one or more internal positions of the sense strand and / or antisense strand, Each nucleotide is attached to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide, respectively; preferably, Linked to the 3' carbon of the previous nucleotide or the corresponding position. Linked to the 5' carbon of the next nucleotide or the corresponding position; or When the corresponding structure is added to the 5' or 3' end of the justice chain and / or antisense chain, One of them is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is added to the 5' end of the sense and / or antisense strands, Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 5' carbon of the 5' terminal nucleotide or at the corresponding position; or when the corresponding structure is added to the 3' end of the sense strand and / or antisense strand. Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position. The method of claim 43, the method comprising including, in the double-stranded RNA, one or more structures independently selected from the following: in, Y is oxygen or sulfur, and Wherein, when the corresponding structure is used to replace nucleotides at one or more internal positions of the sense strand and / or antisense strand, Each nucleotide is attached to the 3' carbon or corresponding position of the previous nucleotide and to the 5' carbon or corresponding position of the next nucleotide, respectively; preferably, Linked to the 3' carbon of the previous nucleotide or the corresponding position. Linked to the 5' carbon of the next nucleotide or the corresponding position; or When the corresponding structure is added to the 5' or 3' end of the justice chain and / or antisense chain, One of them is attached to the 3' or 5' carbon of the terminal nucleotide or the corresponding position, and the other represents attachment to a hydrogen, terminal modification, or terminal protecting group; preferably, when the corresponding structure is added to the 5' end of the sense and / or antisense strands, Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 5' carbon of the 5' terminal nucleotide or at the corresponding position; or when the corresponding structure is added to the 3' end of the sense strand and / or antisense strand. Connected to hydrogen, terminal modification, or terminal protecting group, and Linked to the 3' carbon of the 3' terminal nucleotide or the corresponding position.

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