Nucleotide substitutes with enhanced stability

Abstract

The present invention provides a compound of formula (X), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof. And a compound of formula (X) linked to the 5 '-terminal and / or the 3'-terminal of the oligonucleotide for increasing the stability of the double-stranded RNA or assisting the double-stranded RNA in degrading against exonuclease. The invention also relates to a double-stranded RNA, a vector, a cell, a pharmaceutical composition and a kit comprising the compound of formula (X). # imgabs0 #

Description

Nucleotide substitutes with enhanced stability

[0001] This application claims priority to Chinese application No. 202210744263.8 filed on June 27, 2022, Chinese application No. 202210864592.6 filed on July 21, 2022, and Chinese application No. 202310411631.1 filed on April 17, 2023, which are incorporated herein by reference in their entirety. Field of the Invention

[0002] The present invention belongs to the field of medicine, and in particular relates to nucleotide substitutes with enhanced stability, which are used to improve the stability of double-stranded RNA or help double-stranded RNA resist degradation by exonucleases. Background Art

[0003] RNA interference (RNAi) is a phenomenon in which double-stranded RNA (dsRNA) induces the efficient and specific degradation of target mRNA. Once inside the body, dsRNA is degraded by nucleases, thus affecting the efficiency and duration of RNAi. Therefore, introducing nucleotide substitutions at the ends of RNA molecules, where they are most susceptible to nucleases, may improve their stability.

[0004] Currently, molecules based on morpholine rings or dioxanes have been disclosed, which can improve the stability of double-stranded RNA after being linked to the 3' end of the sense strand. However, there is still a need in the art to develop more nucleotide analogs with enhanced stability.

[0005] Summary of the Invention

[0006] In one aspect, the present invention provides a compound of formula (X), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0007] wherein each group is as defined below.

[0008] In another aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0009] wherein each group is as defined below.

[0010] In another aspect, the present invention provides a compound of formula (Ia), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0011] wherein each group is as defined below.

[0012] In another aspect, the present invention provides a compound of formula (Ib), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0013] wherein each group is as defined below.

[0014] In another aspect, the present invention provides a compound of formula (Ic), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0015] wherein each group is as defined below.

[0016] In another aspect, the present invention provides a compound of formula (II), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0017] wherein each group is as defined below.

[0018] In another aspect, the present invention provides an oligonucleotide comprising one or more compounds of formula (X') at the 5' end and / or the 3' end:

[0019] wherein each group is as defined below.

[0020] In another aspect, the present invention provides an oligonucleotide comprising one or more compounds of formula (I') at the 5' end and / or the 3' end:

[0021] wherein each group is as defined below.

[0022] In another aspect, the present invention provides an oligonucleotide comprising one or more compounds of formula (Ia') at the 5' end and / or the 3' end:

[0023] wherein each group is as defined below.

[0024] In another aspect, the present invention provides an oligonucleotide comprising one or more compounds of formula (Ib') at the 5' end and / or the 3' end:

[0025] wherein each group is as defined below.

[0026] In another aspect, the present invention provides an oligonucleotide comprising one or more compounds of formula (Ic') at the 5' end and / or the 3' end:

[0027] wherein each group is as defined below.

[0028] In another aspect, the present invention provides an oligonucleotide comprising one or more compounds of formula (II') at the 5' end and / or the 3' end:

[0029] wherein each group is as defined below.

[0030] In another aspect, the present invention provides a double-stranded RNA having a sense strand and an antisense strand with fully complementary sequences, each strand having 14 to 30 nucleotides, the nucleotides being linked by phosphate groups, phosphorothioate groups or other linking molecules, wherein the sense strand has the following structure:

[0031] 5'(NT1) n1 -(NT2) n2 -(NT3) n3 3'

[0032] wherein each group is as defined below.

[0033] In another aspect, the present invention provides a vector comprising a nucleotide sequence encoding the aforementioned double-stranded RNA.

[0034] In another aspect, the present invention provides a cell containing the aforementioned double-stranded RNA or the aforementioned vector.

[0035] In another aspect, the present invention provides a pharmaceutical composition comprising the aforementioned double-stranded RNA, the aforementioned vector, or the aforementioned cell, and optionally a pharmaceutically acceptable carrier or excipient.

[0036] In another aspect, the present invention provides a kit comprising the aforementioned double-stranded RNA, the aforementioned vector, or the aforementioned cell.

[0037] The compounds of the present invention can be connected to the 3' end or 5' end of the sense strand to enhance the stability of the RNA double strand. In addition, the compounds of the present invention containing an amino group (including an N-containing heterocycle) can be connected to the 5' end or 3' end of the sense strand to enhance the stability of the RNA double strand and prevent the sense strand from being incorrectly loaded into the Ago2 protein.

[0038] Without being bound by theory, Kumar et al, Chem. Commun., 2019, 55, 5139-5142, mentioned that cations will form a mutual repulsion with Lys-570 and Lys-533 in the Ago2 MID region, resulting in the inability of nucleotide chains containing cations at the 5' end to be loaded into Ago2. Therefore, the introduction of substituted amines at the 5' end of the sense strand can form a cationic structure to reduce the off-target effect caused by the incorrect loading of the sense strand into Ago2 or to improve the loading of the antisense strand.

[0039] Detailed Description of the Invention

[0040] definition

[0041] Chemical definition

[0042] Definitions of specific functional groups and chemical terms are described in more detail below.

[0043] When a numerical range is listed, it is intended to include every value and sub-range within the stated range. For example, "C 1-6 "Alkyl" includes C1, C2, C3, C4, C5, C6, C 1-6 、C 1-5 、C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-5 、C 2-4 、C 2-3 、C 3-6 、C 3-5 、C 3-4 、C 4-6 、C 4-5 and C 5-6 alkyl.

[0044] “C 1-20 "Alkyl" refers to a straight or branched chain saturated hydrocarbon group having 1 to 20 carbon atoms. In some embodiments, C 1-6 Alkyl, C 1-4 Alkyl and C 1-2 Alkyl groups are preferred. 1-6 Examples of alkyl groups include: methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), pentyl (C5), neopentyl (C5), 3-methyl-2-butyl (C5), tert-pentyl (C5), and n-hexyl (C6). The term “C 1-6 "Alkyl" also includes heteroalkyl groups in which one or more (e.g., 1, 2, 3, or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). The alkyl group may be optionally substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. Conventional alkyl abbreviations include: Me(-CH3), Et(-CH2CH3), iPr(-CH(CH3)2), nPr(-CH2CH2CH3), n-Bu(-CH2CH2CH2CH3), or i-Bu(-CH2CH(CH3)2).

[0045] “C 2-20 "Alkenyl" refers to a straight or branched chain hydrocarbon group having 2 to 20 carbon atoms and at least one carbon-carbon double bond. In some embodiments, C2-6 Alkenyl and C 2-4 Alkenyl is preferred. 2-6 Examples of alkenyl groups include ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), pentenyl (C5), pentadienyl (C5), hexenyl (C6), and the like. The term "C 2- "Alkenyl" also includes heteroalkenyl groups in which one or more (e.g., 1, 2, 3, or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). The alkenyl group may be optionally substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0046] “C 2-20 "Alkynyl" refers to a straight or branched chain hydrocarbon group having 2 to 20 carbon atoms, at least one carbon-carbon triple bond, and optionally one or more carbon-carbon double bonds. In some embodiments, C 2-6 Alkynyl and C 2-4 Alkynyl is preferred. 2-6 Examples of alkynyl groups include, but are not limited to, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), pentynyl (C5), hexynyl (C6), and the like. The term "C 2-6 "Alkynyl" also includes heteroalkynyl groups in which one or more (e.g., 1, 2, 3, or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). Alkynyl groups can be optionally substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0047] “C 1-20 Alkylene" and "C 1-6 "Alkylene" refers to the group with the C 1-20 Alkyl and C 1-6 In some embodiments, C 1-4 Alkylene, C 2-4 Alkylene and C 1-3Alkylene is preferred. Unsubstituted alkylene includes, but is not limited to, methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), butylene (-CH2CH2CH2CH2-), pentylene (-CH2CH2CH2CH2CH2-), hexylene (-CH2CH2CH2CH2CH2CH2-), and the like. Exemplary substituted alkylenes, for example, alkylenes substituted with one or more alkyl (methyl) groups, include, but are not limited to, substituted methylene (-CH(CH3)-, -C(CH3)2-), substituted ethylene (-CH(CH3)CH2-, -CH2CH(CH3)-, -C(CH3)2CH2-, -CH2C(CH3) 2- ), substituted propylene (-CH(CH3)CH2CH2-, -CH2CH(CH3)CH2-, -CH2CH2CH(CH3)-, -C(CH3)2CH2CH2-, -CH2C(CH3)2CH2-, -CH2CH2C(CH3)2-), and the like.

[0048] “C 0-6 "Alkylene" refers to the chemical bond and the above-mentioned "C 1-6 "Alkylene".

[0049] "Halo" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

[0050] Therefore, “C 1-20 Halogenated alkyl, "C 1-6 Haloalkyl" and "C 1-4 "Haloalkyl" refers to the above-mentioned "C 1-20 Alkyl", "C 1-6 Alkyl" and "C 1-4 Alkyl", which is substituted by one or more halogen groups. In some embodiments, C 1-4 Halogenated alkyl is particularly preferred, more preferably C 1-2 Haloalkyl. Exemplary haloalkyl groups include, but are not limited to, -CF3, -CH2F, -CHF2, -CHFCH2F, -CH2CHF2, -CF2CF3, -CCl3, -CH2Cl, -CHCl2, 2,2,2-trifluoro-1,1-dimethyl-ethyl, and the like. The haloalkyl group can be substituted at any available point of attachment, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0051] “C 3-8 "Cycloalkyl" refers to a non-aromatic cyclic hydrocarbon group having 3 to 8 ring carbon atoms and zero heteroatoms. In some embodiments, C 3-7 Cycloalkyl and C3-6 Cycloalkyl is particularly preferred, more preferably C 4-6 Cycloalkyl and C 5-6 Cycloalkyl. Cycloalkyl also includes a ring system in which the above-mentioned cycloalkyl ring is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the cycloalkyl ring, and in such a case, the number of carbons continues to represent the number of carbons in the cycloalkyl system. Exemplary cycloalkyls include, but are not limited to, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), and the like. The cycloalkyl group may be optionally substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0052] "3-10 membered heterocyclyl" refers to a saturated or unsaturated radical of a 3- to 10-membered non-aromatic ring system having ring carbon atoms and 1 to 5 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon. In heterocyclyl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, as valence permits. In some embodiments, a 4-10 membered heterocyclyl is preferred, which is a 4 to 10 membered non-aromatic ring system having ring carbon atoms and 1 to 5 ring heteroatoms; in some embodiments, a 3-8 membered heterocyclyl is preferred, which is a 3 to 8 membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms; a 4-8 membered heterocyclyl is preferred, which is a 4 to 8 membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; a 4-7 membered heterocyclyl is preferred, which is a 4 to 7 membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; a 4-6 membered heterocyclyl is preferred, which is a 4 to 6 membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; and a 5-6 membered heterocyclyl is more preferred, which is a 5 to 6 membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms. Heterocyclyl also includes a ring system in which the above-mentioned heterocyclyl ring is fused to one or more cycloalkyl groups, wherein the point of attachment is on the cycloalkyl ring, or a ring system in which the above-mentioned heterocyclyl ring is fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring; and in such a case, the number of ring members continues to represent the number of ring members in the heterocyclyl ring system. Exemplary 3-membered heterocyclyls containing one heteroatom include, but are not limited to, aziridine, oxirane, and thiorenyl. Exemplary 4-membered heterocyclyls containing one heteroatom include, but are not limited to, azetidinyl, oxetane, and thiidine. Exemplary 5-membered heterocyclyls containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to, pyrazolidinyl, dioxolanyl, oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-membered heterocyclic groups containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing one heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithianyl, and dioxanyl. Exemplary 6-membered heterocyclic groups containing three heteroatoms include, but are not limited to, hexahydrotriazinyl. Exemplary 7-membered heterocyclic groups containing one heteroatom include, but are not limited to, azepanyl, oxepanyl, and thienyl.Exemplary 5-membered heterocyclyl groups fused to a C6 aryl ring (also referred to herein as 5,6-bicyclic heterocyclyl groups) include, but are not limited to, dihydroindolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinone, and the like. Exemplary 6-membered heterocyclyl groups fused to a C6 aryl ring (also referred to herein as 6,6-bicyclic heterocyclyl groups) include, but are not limited to, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like. Heterocyclyl groups also include those in which the above-mentioned heterocyclyl groups share one or two atoms with a cycloalkyl, heterocyclyl, aryl, or heteroaryl group to form a bridged or spirocyclic ring. The shared atoms may be carbon or nitrogen atoms, as valence permits. Heterocyclyl groups also include those in which the above-mentioned heterocyclyl and heterocyclyl groups may be optionally substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0053] “C 6-10 "Aryl" refers to a monocyclic or polycyclic (e.g., bicyclic) 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in a cyclic arrangement) having 6-10 ring carbon atoms and zero heteroatoms. In some embodiments, an aryl group has six ring carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, an aryl group has ten ring carbon atoms ("C 10 Aryl also includes ring systems in which an aryl ring as described above is fused to one or more cycloalkyl or heterocyclyl groups, and the point of attachment is on the aryl ring, in which case the number of carbon atoms continues to refer to the number of carbon atoms in the aryl ring system. Aryl groups can be optionally substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0054] "5-14 membered heteroaryl" refers to a group of a 5-14 membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6, 10, or 14 π electrons shared in a cyclic arrangement) having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In heteroaryl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom as long as valence permits. Heteroaryl bicyclic ring systems may include one or more heteroatoms in one or both rings. Heteroaryl also includes ring systems in which the above-mentioned heteroaryl rings are fused to one or more cycloalkyl or heterocyclyl groups, and the point of attachment is on the heteroaryl ring, in which case the number of carbon atoms continues to represent the number of carbon atoms in the heteroaryl ring system. In some embodiments, 5-10 membered heteroaryl is preferred, which is a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms. In other embodiments, 5-6 membered heteroaryls are particularly preferred and are 5-6 membered monocyclic or bicyclic 4n+2 aromatic ring systems having ring carbon atoms and 1-4 ring heteroatoms. Exemplary 5-membered heteroaryls containing one heteroatom include, but are not limited to, pyrrolyl, furanyl, and thienyl. Exemplary 5-membered heteroaryls containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryls containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl (e.g., 1,2,4-oxadiazolyl), and thiadiazolyl. Exemplary 5-membered heteroaryls containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryls containing one heteroatom include, but are not limited to, pyridinyl or pyridonyl. Exemplary 6-membered heteroaryls containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one heteroatom include, but are not limited to, azacycloheptatrienyl, oxepinyl, and thieptatrienyl. Exemplary 5,6-bicyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indanyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, but are not limited to, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. A heteroaryl group can be optionally substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0055] The divalent groups formed by removing another hydrogen from the above-defined alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups are collectively referred to as "subunits". The ring-forming groups such as cycloalkyl, heterocyclyl, aryl and heteroaryl groups are collectively referred to as "cyclyls".

[0056] Alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and the like are defined herein as optionally substituted groups.

[0057] Exemplary substituents on carbon atoms include, but are not limited to, halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR aa 、-ON(R bb )2、-N(R bb )2、-N(R bb )3 + X - 、-N(OR cc )R bb 、-SH、-SR aa 、-SSR cc 、-C(=O)R aa 、-CO2H、-CHO、-C(OR cc )2, -CO2R aa 、-OC(=O)R aa 、-OCO2R aa 、-C(=O)N(R bb )2、-OC(=O)N(R bb )2、-NR bb C(=O)R aa 、-NR bb CO2R aa 、-NR bb C(=O)N(R bb )2、-C(=NR bb )R aa 、-C(=NR bb )OR aa 、-OC(=NR bb )R aa 、-OC(=NR bb )OR aa 、-C(=NR bb )N(R bb )2、-OC(=NR bb )N(R bb )2、-NR bb C(=NR bb )N(R bb )2, -C(=O)NR bb SO2R aa 、-NRbb SO2R aa 、-SO2N(R bb )2, -SO2R aa 、-SO2OR aa 、-OSO2R aa 、-S(=O)R aa 、-OS(=O)R aa 、-Si(R aa )3、-OSi(R aa )3、-C(=S)N(R bb )2, -C(=O)SR aa 、-C(=S)SR aa 、-SC(=S)SR aa 、-SC(=O)SR aa 、-OC(=O)SR aa 、-SC(=O)OR aa 、-SC(=O)R aa 、-P(=O)2R aa 、-OP(=O)2R aa 、-P(=O)(R aa )2、-OP(=O)(R aa )2、-OP(=O)(OR cc )2、-P(=O)2N(R bb )2、-OP(=O)2N(R bb )2、-P(=O)(NR bb )2、-OP(=O)(NR bb )2、-NR bb P(=O)(OR cc )2、-NR bb P(=O)(NR bb )2、-P(R cc )2、-P(R cc )3、-OP(R cc )2、-OP(R cc )3、-B(R aa )2、-B(OR cc )2, -BR aa (OR cc ), alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently replaced by 0, 1, 2, 3, 4 or 5 R dd group substitution;

[0058] Or the two geminal hydrogen atoms on the carbon atom are replaced by groups =O, =S, =NN(R bb)2, =NNR bb C(=O)R aa 、=NNR bb C(=O)OR aa 、=NNR bb S(=O)2R aa 、=NR bb or = NOR cc replace;

[0059] R aa Each of R is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R aa The groups are combined to form a heterocyclyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl group is independently replaced by 0, 1, 2, 3, 4 or 5 R dd group substitution;

[0060] R bb Each of the following is independently selected from: hydrogen, -OH, -OR aa 、-N(R cc )2, -CN, -C(=O)R aa 、-C(=O)N(R cc )2, -CO2R aa 、-SO2R aa 、-C(=NR cc )OR aa 、-C(=NR cc )N(R cc )2、-SO2N(R cc )2, -SO2R cc 、-SO2OR cc 、-SOR aa 、-C(=S)N(R cc )2, -C(=O)SR cc 、-C(=S)SR cc 、-P(=O)2R aa 、-P(=O)(R aa )2、-P(=O)2N(R cc )2、-P(=O)(NR cc )2, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R bb The groups are combined to form a heterocyclyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl group is independently replaced by 0, 1, 2, 3, 4 or 5 R dd group substitution;

[0061] R ccEach of R is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R cc The groups are combined to form a heterocyclyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl group is independently replaced by 0, 1, 2, 3, 4 or 5 R dd group substitution;

[0062] R dd Each of the is independently selected from: halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR ee 、-ON(R ff )2、-N(R ff )2,、-N(R ff )3 + X - 、-N(OR ee )R ff 、-SH、-SR ee 、-SSR ee 、-C(=O)R ee 、-CO2H、-CO2R ee 、-OC(=O)R ee 、-OCO2R ee 、-C(=O)N(R ff )2、-OC(=O)N(R ff )2、-NR ff C(=O)R ee 、-NR ff CO2R ee 、-NR ff C(=O)N(R ff )2、-C(=NR ff )OR ee 、-OC(=NR ff )R ee 、-OC(=NR ff )OR ee 、-C(=NR ff )N(R ff )2、-OC(=NR ff )N(R ff )2、-NR ff C(=NR ff )N(R ff )2、-NR ff SO2R ee 、-SO2N(R ff )2, -SO2R ee 、-SO2OR ee 、-OSO2R ee、-S(=O)R ee 、-Si(R ee )3、-OSi(R ee )3、-C(=S)N(R ff )2, -C(=O)SR ee 、-C(=S)SR ee 、-SC(=S)SR ee 、-P(=O)2R ee 、-P(=O)(R ee )2、-OP(=O)(R ee )2、-OP(=O)(OR ee )2, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R gg Group substitution, or two geminal R dd Substituents may combine to form =O or =S;

[0063] R ee Each of R is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclyl and heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently replaced by 0, 1, 2, 3, 4 or 5 R gg group substitution;

[0064] R ff Each of R is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R ff The groups are combined to form a heterocyclic or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl and heteroaryl group is independently replaced by 0, 1, 2, 3, 4 or 5 R gg group substitution;

[0065] R gg Each of the independently: halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OC 1-6 Alkyl, -ON(C 1-6 Alkyl)2, -N(C 1-6 Alkyl)2, -N(C 1-6 Alkyl)3 + X - 、-NH(C 1-6 Alkyl)2 + X - 、-NH2(C 1-6 alkyl) + X - 、-NH3+ X - 、-N(OC 1-6 Alkyl)(C 1-6 Alkyl), -N(OH)(C 1-6 Alkyl), -NH(OH), -SH, -SC 1-6 Alkyl, -SS(C 1-6 alkyl), -C(=O)(C 1-6 alkyl), -CO2H, -CO2(C 1-6 alkyl), -OC(=O)(C 1-6 Alkyl), -OCO2(C 1-6 alkyl), -C(=O)NH2, -C(=O)N(C 1-6 alkyl)2, -OC(=O)NH(C 1-6 alkyl), -NHC(=O)(C 1-6 Alkyl), -N(C 1-6 alkyl)C(=O)(C 1-6 Alkyl), -NHCO2(C 1-6 alkyl), -NHC(=O)N(C 1-6 Alkyl)2, -NHC(=O)NH(C 1-6 alkyl), -NHC(=O)NH2, -C(=NH)O(C 1-6 alkyl), -OC(=NH)(C 1- 6 alkyl), -OC(=NH)OC 1-6 Alkyl, -C(=NH)N(C 1-6 Alkyl)2, -C(=NH)NH(C 1-6 alkyl), -C(=NH)NH2, -OC(=NH)N(C 1-6 Alkyl)2, -OC(NH)NH(C 1-6 alkyl), -OC(NH)NH2, -NHC(NH)N(C 1-6 Alkyl)2, -NHC(=NH)NH2, -NHSO2(C 1-6 Alkyl), -SO2N(C 1-6 Alkyl)2, -SO2NH(C 1-6 alkyl), -SO2NH2, -SO2C 1-6 Alkyl, -SO2OC 1-6 Alkyl, -OSO2C 1-6 Alkyl, -SOC 1-6 Alkyl, -Si(C 1-6 alkyl)3, -OSi(C 1-6 alkyl)3, -C(=S)N(C 1-6 alkyl)2、C(=S)NH(C 1-6alkyl), C(=S)NH2, -C(=O)S(C 1-6 alkyl), -C(=S)SC 1-6 Alkyl, -SC(=S)SC 1-6 Alkyl, -P(=O)2(C 1-6 alkyl), -P(=O)(C 1- 6 alkyl) 2, -OP(=O)(C 1-6 alkyl)2, -OP(=O)(OC 1-6 Alkyl)2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, C6-C 10 Aryl, C3-C7 heterocyclic, C5-C 10 heteroaryl; or two geminal R gg Substituents may combine to form =O or =S; wherein X - For the counter ion.

[0066] Exemplary substituents on nitrogen atoms include, but are not limited to, hydrogen, -OH, -OR aa 、-N(R cc )2, -CN, -C(=O)R aa 、-C(=O)N(R cc )2, -CO2R aa 、-SO2R aa 、-C(=NR bb )R aa 、-C(=NR cc )OR aa 、-C(=NR cc )N(R cc )2、-SO2N(R cc )2, -SO2R cc 、-SO2OR cc 、-SOR aa 、-C(=S)N(R cc )2, -C(=O)SR cc 、-C(=S)SR cc 、-P(=O)2R aa 、-P(=O)(R aa )2、-P(=O)2N(R cc )2、-P(=O)(NR cc )2, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R attached to the nitrogen atom ccThe groups are combined to form a heterocyclic or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl and heteroaryl group is independently replaced by 0, 1, 2, 3, 4 or 5 R dd group substituted, and wherein R aa 、R bb 、R cc and R dd As mentioned above.

[0067] Other definitions

[0068] The term "siRNA" herein refers to a class of double-stranded RNA molecules that can mediate the silencing of a target RNA (e.g., mRNA, e.g., a transcript of a gene encoding a protein) that is complementary thereto. siRNA is typically double-stranded, comprising an antisense strand complementary to the target RNA and a sense strand complementary to the antisense strand. For convenience, such mRNA is also referred to herein as mRNA to be silenced. Such a gene is also referred to as a target gene. Typically, the RNA to be silenced is an endogenous gene or a pathogen gene. In addition, RNA (e.g., tRNA) and viral RNA other than mRNA can also be targeted.

[0069] The term "antisense strand" refers to a strand of an siRNA that includes a region that is fully or substantially complementary to a target sequence. The term "sense strand" refers to a strand of an siRNA that includes a region that is substantially complementary to a region that is the antisense strand as that term is defined herein.

[0070] The term "complementary region" refers to a region on the antisense strand that is completely or substantially complementary to the target mRNA sequence. In the case where the complementary region is not completely complementary to the target sequence, mispairing can be located in the interior or terminal regions of the molecule. Typically, the most tolerated mispairing is located in the terminal regions, for example, within 5, 4, 3, 2 or 1 nucleotides at 5' and / or 3' ends. The antisense strand portion that is most sensitive to mispairing is referred to as a "seed region." For example, in a siRNA comprising a 19nt chain, the 19th position (from 5' to 3') can tolerate some mispairing.

[0071] The term "complementary" refers to the ability of a first polynucleotide to hybridize to a second polynucleotide under certain conditions, such as stringent conditions. For example, stringent conditions may include 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mM EDTA at 50° C. or 70° C. for 12-16 hours. In terms of meeting the above requirements relative to their ability to hybridize, "complementary" sequences may also include or be completely formed from non-Watson-Crick base pairs and / or base pairs formed from non-natural and modified nucleotides. Such non-Watson-Crick base pairs include, but are not limited to, G:U wobble base pairing or Hoogstein base pairing.

[0072] A polynucleotide that is "at least partially complementary" or "substantially complementary" to a messenger RNA (mRNA) refers to a polynucleotide that is substantially complementary to a contiguous portion of the mRNA of interest. For example, a polynucleotide is complementary to at least a portion of a PCSK9 mRNA if the sequence is substantially complementary to a non-interrupted portion of an mRNA encoding PCSK9. The terms "complementary," "fully complementary," and "substantially complementary" as used herein can be used with respect to base pairing between the sense and antisense strands of an siRNA, or between the antisense strand of an siRNA agent and a target sequence.

[0073] "shRNA" stands for short hairpin RNA. shRNA consists of two short inverted repeats. When cloned into an shRNA expression vector, the two short inverted repeats are separated by a stem-loop sequence, forming a hairpin structure controlled by a Pol III promoter. Five to six Ts are then attached to serve as a transcriptional terminator for RNA polymerase III.

[0074] A "nucleoside" is a compound composed of a purine or pyrimidine base and ribose or deoxyribose; a "nucleotide" is a compound composed of a purine or pyrimidine base, ribose or deoxyribose, and phosphate; an "oligonucleotide" is a nucleic acid molecule (RNA or DNA) having, for example, fewer than 100, 200, 300, or 400 nucleotides in length.

[0075] "Bases" are the fundamental building blocks of nucleosides, nucleotides, and nucleic acids. They contain nitrogen and are also called "nitrogenous bases." Unless otherwise specified, the capital letters A, U, T, G, and C represent the bases of nucleotides, representing adenine, uracil, thymine, guanine, and cytosine, respectively.

[0076] The "modification" of nucleotides herein includes, but is not limited to, methoxy modification, fluorination, phosphorothioate linkage, or conventional protecting group protection. For example, the fluorination-modified nucleotide refers to a nucleotide in which the hydroxyl group at the 2' position of the ribose group of the nucleotide is replaced by fluorine, and the methoxy-modified nucleotide refers to a nucleotide in which the 2'-hydroxyl group of the ribose group is replaced by a methoxy group.

[0077] " modified nucleotide " herein includes but is not limited to 2 '-O-methyl modified nucleotides, 2 '-fluoro modified nucleotides, 2 '-deoxy-modified nucleotides, inosine ribonucleotides, abasic nucleotides, reverse abasic deoxyribonucleotides, nucleotides comprising thiophosphate groups, vinyl phosphate modified nucleotides, locked nucleotides, 2 '-amino-modified nucleotides, 2 '-alkyl-modified nucleotides, morpholino nucleotides, phosphoramidates, the non-natural bases comprising nucleotides and the terminal nucleotides, deoxyribonucleotides or conventional protecting group protections on a cholesterol derivative or a dodecanoic acid didecylamide group. For example, the 2 '-fluoro modified nucleotides refer to nucleotides in which the hydroxyl group at the ribose group 2 ' position is replaced by fluorine. The 2 '-deoxy-modified nucleotides refer to nucleotides in which the 2 '-hydroxyl group at the ribose group is replaced by methoxy and formed.

[0078] "Ligand moiety" refers to a chemical moiety conjugated to an siRNA that is capable of altering the distribution, targeting, or lifetime of the siRNA. In preferred embodiments, such a ligand provides enhanced affinity for a selected target (e.g., a molecule, a cell or cell type, a compartment (e.g., a cell or organ compartment, a tissue, an organ, or a region of the body) compared to, for example, an siRNA in the absence of such a ligand.

[0079] " reactive phosphorus group " refers to the phosphorus-containing group contained in the nucleotide unit or in the nucleotide analog unit, and it can react with the hydroxyl or amine group contained in another molecule, especially in another nucleotide unit or in another nucleotide analog by nucleophilic attack reaction.Usually, such reaction produces the ester type internucleoside bond that the first nucleotide unit or the first nucleotide analog unit is connected to the second nucleotide unit or the second nucleotide analog unit.Reactive phosphorus group can be selected from phosphoramidites, H-phosphonates, alkyl-phosphonates, phosphate or phosphate mimetics, including but not limited to: natural phosphate, thiophosphate, phosphorodithioate, borane phosphate, borane thiophosphate, phosphonate, halogen-substituted phosphonate and phosphate, phosphoramidate, phosphodiester, phosphotriester, thiophosphate diester, thiophosphate triester, diphosphate and triphosphate.

[0080] A "protecting group" refers to any atom or group of atoms added to a molecule to prevent an existing group in the molecule from undergoing an undesirable chemical reaction. A "protecting group" may be an unstable chemical moiety known in the art that is used to protect reactive groups, such as hydroxyl, amino, and thiol groups, to prevent undesirable or inappropriate reactions during chemical synthesis. Protecting groups are typically used selectively and / or orthogonally to protect sites during reactions at other reactive sites and can then be removed to leave the unprotected group intact or available for further reactions.

[0081] A non-limiting list of protecting groups includes benzyl; substituted benzyl; alkylcarbonyl and alkoxycarbonyl (e.g., tert-butyloxycarbonyl (BOC), acetyl, or isobutyryl); arylalkylcarbonyl and arylalkoxycarbonyl (e.g., benzyloxycarbonyl); substituted methyl ethers (e.g., methoxymethyl ether); substituted ethyl ethers; substituted benzyl ethers; tetrahydropyranyl ethers; silyl (e.g., trimethylsilyl, triethylsilyl, triisopropylsilyl, tert-butyldimethylsilyl, tri-isopropylsilyloxymethyl, [2-(trimethylsilyl)ethoxy] methyl or tert-butyldiphenylsilyl); esters (e.g., benzoate); carbonates (e.g., methoxymethyl carbonate); sulfonates (e.g., tosylate or mesylate); acyclic ketals (e.g., dimethyl acetal); cyclic ketals (e.g., 1,3-dioxane, 1,3-dioxolane, and those described herein); acyclic acetals; cyclic acetals (e.g., those described herein); acyclic hemiacetals; cyclic hemiacetals; cyclic dithioketals (e.g., 1,3-dithiane or 1,3-dithiolane); orthoesters (e.g., those described herein), and triarylmethyl groups (e.g., trityl; monomethoxytrityl (MMTr); 4,4′-dimethoxytrityl (DMTr); 4,4′,4″-trimethoxytrityl (TMTr); and those described herein). Preferred protecting groups are selected from acetyl (Ac), benzoyl (Bzl), benzyl (Bn), isobutyryl (iBu), phenylacetyl, benzyloxymethyl acetal (BOM), β-methoxyethoxymethyl ether (MEM), methoxymethyl ether (MOM), p-methoxybenzyl ether (PMB), methylthiomethyl ether, neopentyl (Vl), methylthiomethyl ether, ... Valeryl (Piv), tetrahydropyranyl (THP), triphenylmethyl (Trt), methoxytrityl [(4-methoxyphenyl)diphenylmethyl] (MMT), dimethoxytrityl, [bis-(4-methoxyphenyl)phenylmethyl] (DMT), trimethylsilyl ether (TMS), tert-butyldimethylsilyl ether (TBDMS), tri-iso-propylsilyloxymethyl ether (TOM), tri-isopropylsilyl ether (TIPS), methyl ether, ethoxyethyl ether (EE) N,N-dimethylformamidine and 2-cyanoethyl (CE).

[0082] A "hydroxyl protecting group" is a group that protects the hydroxyl group from chemical reactions and can be removed under specific conditions to restore the hydroxyl group. These groups primarily include silane-type protecting groups, acyl-type protecting groups, or ether-type protecting groups, with the following being preferred:

[0083] Trimethylsilyl (TMS), triethylsilyl (TES), dimethylisopropylsilyl (DMIPS), diethylisopropylsilyl (DEIPS), tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), triisopropylsilyl (TIPS), acetyl (Ac), chloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl (TFA), benzoyl, p-methoxybenzoyl, 9-fluorenylmethoxycarbonyl (Fmoc), allyloxycarbonyl (Al loc), 2,2,2-trichloroethoxycarbonyl (Troc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), benzyl (Bn), p-methoxybenzyl (PMB), allyl, triphenylmethyl (Tr), bis-p-methoxytrityl (DMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzyloxymethyl (PMBM).

[0084] As used herein, the term "pharmaceutically acceptable salt" refers to those carboxylate salts, amino acid addition salts of the compounds of the present invention that are suitable for use in contact with patient tissues within the scope of sound medical judgment, do not produce undue toxicity, irritation, allergic response, etc., are commensurate with a reasonable benefit / risk ratio, and are effective for their intended use, including (where possible) zwitterionic forms of the compounds of the present invention.

[0085] The present invention includes tautomers, which are functional isomers produced by the rapid movement of an atom in two positions in a molecule. Compounds that exist in different tautomeric forms are not limited to any specific tautomer, but are intended to cover all tautomeric forms.

[0086] The compounds of the present invention may include one or more asymmetric centers and may therefore exist in a variety of stereoisomeric forms, for example, enantiomers and / or diastereomeric forms. For example, the compounds of the present invention may be individual enantiomers, diastereomers, or geometric isomers (e.g., cis and trans isomers), or may be in the form of mixtures of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers may be separated from the mixture by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers may be prepared by asymmetric synthesis.

[0087] The present invention also includes isotopically labeled compounds (isotopic variants) which are identical to those described in formula (I) but for which one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be introduced into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, for example 2 H. 3 H. 13 C. 11 C. 14 C. 15 N. 18 O. 17 O. 31 P. 32 P. 35 S. 18 F and 36 Cl. Compounds of the present invention containing the above-mentioned isotopes and / or other isotopes of other atoms, their prodrugs and pharmaceutically acceptable salts of the compounds or prodrugs are within the scope of the present invention. Certain isotopically labeled compounds of the present invention, such as those in which radioactive isotopes (e.g. 3 H and 14 C) can be used in drug and / or substrate tissue distribution assays. 3 H and carbon-14, i.e. 14 C isotopes are particularly preferred because they are easy to prepare and detect. 2 H, because greater metabolic stability can provide therapeutic benefits, such as prolonged in vivo half-life or reduced dosage requirements, and thus may be preferred in some cases. Isotopically labeled compounds of formula (I) of the present invention and their prodrugs can generally be prepared by substituting readily available isotopically labeled reagents for non-isotopically labeled reagents when carrying out the processes disclosed in the following schemes and / or the Examples and Preparations.

[0088] Compounds of the present invention

[0089] The present invention specifically relates to a compound of formula (X), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0090] in,

[0091] X1 is a chemical bond, O or S;

[0092] X2 is a chemical bond or NR1;

[0093] Y is a chemical bond or [C(R a )(R b )] 1-4 ;

[0094] R1 is selected from C 1-20 Alkyl, C 1-20 Halogenated alkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl, 5-14 membered heteroaryl or -[C(O)] p -C 1-20 Alkylene-(OCH2CH2) n R x , which are optionally replaced by one or more radicals selected from halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl or 5-14 membered heteroaryl; wherein R1 is optionally deuterated, up to fully deuterated;

[0095] R2 is selected from H, D or OL2;

[0096] R is selected from H, D, -L-OR', -L-NR"R"', R b or -C(OL2)(R c )(R d ), which are optionally deuterated, up to fully deuterated;

[0097] L1 and L2 are selected from H, a reactive phosphorus group or a protecting group;

[0098] R a 、R b 、R c and R d independently selected from H, D, halogen, CN, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -LC 3-8 Cycloalkyl or -L-4-7 membered heterocyclic group; wherein R a 、R b 、R c and R d Optionally deuterated, up to fully deuterated;

[0099] Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated;

[0100] R', R" and R'" are independently selected from H, -C 1-6 Alkylene-OH, -C 1-6 Alkylene-NH2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2- 6 alkenyl, C 2-6 Alkynyl, -C 0-6 Alkylene-C 3-8 Cycloalkyl, -C 0-6 Alkylene-3-10 membered heterocyclic group, -C 0-6 Alkylene-C 6-10 Aryl or -C 0- 6-alkylene-5-14 membered heteroaryl, wherein the above groups are optionally substituted by one or more selected from halogen, CN, NO2, OH, NH2, C 1-6 Alkyl or C 1-6 or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group;

[0101] R x Selected from H, D, OH, OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl; wherein R x Optionally deuterated, up to fully deuterated;

[0102] n=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0103] p = 0 or 1;

[0104] R s Selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0105] m = 0, 1, 2, 3, or 4;

[0106] Provided that one of R and R2 is an OL2 group.

[0107] X1

[0108] In one embodiment, X1 is a chemical bond; in another embodiment, X1 is O; in another embodiment, X1 is S.

[0109] X2

[0110] In one embodiment, X2 is a chemical bond; in another embodiment, X2 is NR1.

[0111] Y

[0112] In one embodiment, Y is a chemical bond; in another embodiment, Y is [C(R a )(R b )] 1-4 , such as C(R a )(R b ),[C(R a )(R b )]2、[C(R a )(R b )]3 or [C(R a )(R b )]4, preferably C(R a )(R b ).

[0113] R1

[0114] In one embodiment, R1 is C 1-20 Alkyl; in another embodiment, R1 is C 1-20 In another embodiment, R1 is C 2-20 In another embodiment, R1 is C 2-20 Alkynyl; in another embodiment, R1 is C 3-8 Cycloalkyl, preferably C 5-6 In another embodiment, R1 is a 3-10 membered heterocyclyl; in another embodiment, R1 is C 6- 10 Aryl; in another embodiment, R1 is 5-14 membered heteroaryl; in another embodiment, R1 is -[C(O)] p -C 1-20 Alkylene-(OCH2CH2) n R x In another embodiment, R1 is selected from C 1-20 Alkyl, C 1-20 Halogenated alkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, C 3- 8-cycloalkyl, 3-10-membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl; In another embodiment, R1 is selected from C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl; In another embodiment, R1 is selected from C3-8 Cycloalkyl or 3-10 membered heterocyclic group.

[0115] In a more specific embodiment, R1 is optionally replaced by one or more radicals selected from halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 In another more specific embodiment, R1 is optionally substituted by one or more groups selected from halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl or C 1-6 In another more specific embodiment, R1 is optionally substituted by one or more groups selected from halogen, CN, OR', SR', NR"R"', C 1-6 Alkyl or C 1-6 In another more specific embodiment, R1 is optionally substituted with deuterated alkyl, up to fully deuterated alkyl.

[0116] R2

[0117] In one embodiment, R2 is H; in another embodiment, R2 is D; in another embodiment, R2 is OL2.

[0118] R

[0119] In one embodiment, R is H; in another embodiment, R is D; in another embodiment, R is -L-OR'; in another embodiment, R is -L-NR"R"'; in another embodiment, R is R b In another embodiment, R is -C(OL2)(R c )(R d ); In another embodiment, R is selected from H, D, -L-OR', -L-NR"R"' or R b In another embodiment, one of R is -C(OL2)(R c )(R d ).

[0120] In a more specific embodiment, R is optionally deuterated, up to fully deuterated.

[0121] L1 and L2

[0122] In one embodiment, L1 and L2 are H; in another embodiment, L1 and L2 are reactive phosphorus groups; in another embodiment, L1 and L2 are protecting groups; in another embodiment, one of L1 and L2 is a reactive phosphorus group and the other is a protecting group.

[0123] R a 、R b 、R c and R d

[0124] In one embodiment, R a 、R b 、R c and R d is H; in another embodiment, R a 、R b 、R c and R d is D; in another embodiment, R a 、R b 、R c and R d is halogen; in another embodiment, R a 、R b 、R c and R d is CN; in another embodiment, R a 、R b 、R c and R d is -L-OR'; in another embodiment, R a 、R b 、R c and R d is -L-NR"R"'; in another embodiment, R a 、R b 、R c and R d C 1-6 Alkyl; in another embodiment, R a 、R b 、R c and R d C 1-6 haloalkyl; in another embodiment, R a 、R b 、R c and R d C 2-6alkenyl; in another embodiment, R a 、R b 、R c and R d C 2-6 Alkynyl; in another embodiment, R a 、R b 、R c and R d For-LC 3-8 Cycloalkyl; in another embodiment, R a 、R b 、R c and R d is -L-4-7 membered heterocyclyl; in another embodiment, R a 、R b 、R c and R d Independently selected from H, D, halogen, CN, -C 0-6 Alkyl-OR', -C 0-6 Alkyl-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl; in another embodiment, R a 、R b 、R c and R d Independently selected from H, D, halogen, CN, C 1-6 Alkyl or C 1-6 Halogenated alkyl.

[0125] In a more specific embodiment, R a 、R b 、R c and R d Optionally deuterated, up to fully deuterated.

[0126] L

[0127] In one embodiment, L is a chemical bond; in another embodiment, L is C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated.

[0128] R', R" and R"'

[0129] In one embodiment, R', R", and R'" are H; in another embodiment, R', R", and R'" are -C 1-6 Alkylene-OH; in another embodiment, R', R" and R"' are -C 1-6 Alkylene-NH2; in another embodiment, R', R" and R"' are C1- 6 alkyl; in another embodiment, R', R" and R"' are C 1-6 haloalkyl; in another embodiment, R', R" and R"' are C 2-6 In another embodiment, R', R" and R"' are C 2-6 Alkynyl; in another embodiment, R', R" and R"' are -C 0-6 Alkylene-C 3-8 Cycloalkyl; in another embodiment, R', R" and R"' are -C 0-6 Alkylene-3-10 membered heterocyclyl; in another embodiment, R', R" and R"' are -C 0-6 Alkylene-C 6-10 Aryl; in another embodiment, R', R" and R"' are -C 0- 6-alkylene-5-14 membered heteroaryl; in another embodiment, R", R"' and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group.

[0130] In a more specific embodiment, R', R" and R'" are optionally replaced by one or more selected from halogen, CN, NO2, OH, NH2, C 1-6 Alkyl or C 1-6 The haloalkyl group is substituted.

[0131] R x

[0132] In one embodiment, R x is H; in another embodiment, R x is D; in another embodiment, R x is OH; in another embodiment, R x For OC 1-6 Alkyl; in another embodiment, R x C 1-6 Alkyl; in another embodiment, R x C 1-6 haloalkyl; in another embodiment, R x C 2-6 alkenyl; in another embodiment, R x C 2-6 Alkynyl; in another embodiment, R x C 3-8 Cycloalkyl; in another embodiment, R x is a 3-10 membered heterocyclyl; in another embodiment, R x C 6-10 Aryl; in another embodiment, R xIt is a 5-14 membered heteroaryl group.

[0133] In a more specific embodiment, R x Optionally deuterated, up to fully deuterated.

[0134] n

[0135] In one embodiment, n=0; in another embodiment, n=1; in another embodiment, n=2; in another embodiment, n=3; in another embodiment, n=4; in another embodiment, n=5; in another embodiment, n=6; in another embodiment, n=7; in another embodiment, n=8; in another embodiment, n=9; in another embodiment, n=10.

[0136] p

[0137] In one embodiment, p=0; in another embodiment, p=1.

[0138] R

[0139] In one embodiment, R s is H; in another embodiment, R s is D; in another embodiment, R s C 1-6 Alkyl; in another embodiment, R s C 1-6 Halogenated alkyl.

[0140] In a more specific embodiment, R s Optionally deuterated, up to fully deuterated.

[0141] m

[0142] In one embodiment, m=0; in another embodiment, m=1; in another embodiment, m=2; in another embodiment, m=3; in another embodiment, m=4.

[0143] Any technical solution or any combination thereof in any of the above specific embodiments can be combined with any technical solution or any combination thereof in other specific embodiments. For example, any technical solution or any combination thereof of X1 can be combined with X2, Y, R1, R2, R, L1, L2, R a 、R b 、R c 、R d ,L,R',R",R"',R x 、R sThe present invention is intended to include combinations of all these technical solutions, which are not listed one by one due to space limitations.

[0144] The present invention also provides a vector comprising a nucleotide sequence encoding the siRNA of the present invention. The vector of the present invention is capable of amplifying or expressing the nucleotide sequence encoding the siRNA of the present invention linked thereto.

[0145] For example, siRNA targeting the PCSK9 gene can be expressed from a transcription unit inserted into a DNA or RNA vector. Expression can be short-lived (a few hours to a few weeks) or continuous (a few weeks to a few months or longer), depending on the specific construct and target tissue or cell type used. The coding nucleotides of the siRNA can be introduced into a linear construct, a circular plasmid or a viral vector. The nucleotides of the siRNA can be integrated into the cell genome for stable expression, or expressed in an extrachromosomal stable inheritance. In general, siRNA expression vectors are typically DNA plasmids or viral vectors.

[0146] Viral vector systems containing siRNA coding sequences include, but are not limited to: (a) adenoviral vectors; (b) retroviral vectors; (c) adeno-associated viral vectors; (d) herpes simplex virus vectors; (e) SV40 vectors; (f) polyoma virus vectors; (g) papilloma virus vectors; (h) picornavirus vectors; (i) poxvirus vectors; and (j) helper virus-dependent adenovirus or gut-free adenovirus.

[0147] The present invention also provides a cell containing the siRNA or vector of the present invention, wherein the siRNA or vector of the present invention can be transcribed in the cell.

[0148] The present invention specifically relates to the following technical solutions:

[0149] 1. A compound of formula (X), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0150] in,

[0151] X1 is a chemical bond, O or S;

[0152] X2 is a chemical bond or NR1;

[0153] Y is a chemical bond or [C(R a )(R b )] 1-4 ;

[0154] R1 is selected from C 1-20 Alkyl, C 1-20 Halogenated alkyl, C2-20 Alkenyl, C 2-20 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl, 5-14 membered heteroaryl or -[C(O)] p -C 1-20 Alkylene-(OCH2CH2) n R x , which are optionally replaced by one or more radicals selected from halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl or 5-14 membered heteroaryl; wherein R1 is optionally deuterated, up to fully deuterated;

[0155] R2 is selected from H, D or OL2;

[0156] R is selected from H, D, -L-OR', -L-NR"R"', R b or -C(OL2)(R c )(R d ), which are optionally deuterated, up to fully deuterated;

[0157] L1 and L2 are selected from H, a reactive phosphorus group or a protecting group;

[0158] R a 、R b 、R c and R d independently selected from H, D, halogen, CN, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -LC 3-8 Cycloalkyl or -L-4-7 membered heterocyclic group; wherein R a 、R b 、R c and R d Optionally deuterated, up to fully deuterated;

[0159] Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated;

[0160] R', R" and R'" are independently selected from H, -C1-6 Alkylene-OH, -C 1-6 Alkylene-NH2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2- 6 alkenyl, C 2-6 Alkynyl, -C 0-6 Alkylene-C 3-8 Cycloalkyl, -C 0-6 Alkylene-3-10 membered heterocyclic group, -C 0-6 Alkylene-C 6-10 Aryl or -C 0- 6-alkylene-5-14 membered heteroaryl, wherein the above groups are optionally substituted by one or more selected from halogen, CN, NO2, OH, NH2, C 1-6 Alkyl or C 1-6 or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group;

[0161] R x Selected from H, D, OH, OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl; wherein R x Optionally deuterated, up to fully deuterated;

[0162] n=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0163] p = 0 or 1;

[0164] R s Selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0165] m = 0, 1, 2, 3, or 4;

[0166] Provided that one of R and R2 is an OL2 group.

[0167] 2. The compound of formula (X) of technical solution 1, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein X1 is a chemical bond or O, preferably O.

[0168] 3. The compound of formula (X) according to technical solution 1 or 2, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein X2 is a chemical bond.

[0169] 4. The compound of formula (X) according to technical solution 1 or 2, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein X2 is NR1.

[0170] 5. The compound of formula (X) according to any one of technical solutions 1 to 4, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein Y is a chemical bond.

[0171] 6. The compound of formula (X) according to any one of technical solutions 1 to 4, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein Y is [C(R a )(R b )] 1-4 , preferably C(R a )(R b ).

[0172] 7. The compound of formula (X) according to any one of technical solutions 1 to 6, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R1 is selected from C 1-20 Alkyl, C 1-20 Halogenated alkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl, preferably selected from C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl, more preferably selected from C 3-8 Cycloalkyl or 3-10 membered heterocyclic group, especially C 5-6 Cycloalkyl.

[0173] 8. The compound of formula (X) of technical solution 7, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R1 is optionally replaced by one or more halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl or C 1-6 The haloalkyl group is substituted, preferably by one or more groups selected from halogen, CN, OR', SR', NR"R"', C 1-6 Alkyl or C 1-6 wherein R1 is optionally substituted with deuterium, up to full deuteration.

[0174] 9. The compound of formula (X) according to any one of technical solutions 1 to 8, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R2 is selected from H or D.

[0175] 10. The compound of formula (X) according to any one of technical solutions 1 to 8, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R2 is OL2.

[0176] 11. The compound of formula (X) according to any one of technical solutions 1 to 10, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R is selected from H, D, -L-OR', -L-NR"R"' or R b , which is optionally deuterated, up to fully deuterated.

[0177] 12. The compound of formula (X) according to any one of technical solutions 1 to 10, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein one of R is -C(OL2)(R c )(R d ), which are optionally deuterated, up to fully deuterated.

[0178] 13. The compound of formula (X) according to any one of technical solutions 1 to 12, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein L1 and L2 are H.

[0179] 14. A compound of formula (X) according to any one of technical solutions 1 to 12, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein one of L1 and L2 is a reactive phosphorus group, preferably a phosphoramidite (preferably -P(OCH2CH2CN)(N(iPr)2)), an H-phosphonate, an alkyl-phosphonate, a phosphate or a phosphate mimetic, such as a natural phosphate, a thiophosphate, a dithiophosphate, a boranephosphate, a boranephosphothioate, a phosphonate, a halogen-substituted phosphonate and a phosphate, an aminophosphate, a phosphodiester, a phosphotriester, a thiophosphate diester, a thiophosphate triester, a diphosphate or a triphosphate.

[0180] 15. The compound of formula (X) according to any one of technical solutions 1 to 12, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein one of L1 and L2 is a protecting group, preferably a hydroxy protecting group, such as trimethylsilyl (TMS), triethylsilyl (TES), dimethylisopropylsilyl (DMIPS), diethylisopropylsilyl (DEIPS), tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), triisopropylsilyl (TIPS), acetyl (Ac), chloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl (TFA), Benzoyl, p-methoxybenzoyl, 9-fluorenylmethoxycarbonyl (Fmoc), allyloxycarbonyl (Alloc), 2,2,2-trichloroethoxycarbonyl (Troc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), benzyl (Bn), p-methoxybenzyl (PMB), allyl, triphenylmethyl (Tr), bis-p-methoxytrityl (DMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzyloxymethyl (PMBM).

[0181] 16. The compound of formula (X) according to any one of technical solutions 1 to 15, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R a 、R b 、R c and R d Independently selected from H, D, halogen, CN, -C 0-6 Alkyl-OR', -C 0-6 Alkyl-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, preferably H, D, halogen, CN, C 1-6 Alkyl or C 1-6 Haloalkyl; wherein R a 、R b 、R c and R d Optionally deuterated, up to fully deuterated.

[0182] 17. The compound of formula (X) according to any one of technical solutions 1 to 16, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, which is selected from the following general formula:

[0183] Wherein each group is as defined in technical solutions 1-16.

[0184] 18. The compound of formula (X) of technical solution 17, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, which is a compound of formula (I):

[0185] in,

[0186] X1 is a chemical bond, O or S;

[0187] R1 is selected from C 1-20 Alkyl, C 1-20 Halogenated alkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl, 5-14 membered heteroaryl or -[C(O)] p -C 1-20 Alkylene-(OCH2CH2) n R x , which are optionally replaced by one or more radicals selected from halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl or 5-14 membered heteroaryl; wherein R1 is optionally deuterated, up to fully deuterated;

[0188] Any of R is -C(OL2)(R c )(R d ), the other two are R b ;

[0189] L1 and L2 are selected from H, a reactive phosphorus group or a protecting group;

[0190] R a 、R b 、R c and R d independently selected from H, D, halogen, CN, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -LC 3-8 Cycloalkyl or -L-4-7 membered heterocyclic group; wherein R a 、R b 、R c and R dOptionally deuterated, up to fully deuterated;

[0191] Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated;

[0192] R', R" and R'" are independently selected from H, -C 1-6 Alkylene-OH, -C 1-6 Alkylene-NH2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2- 6 alkenyl, C 2-6 Alkynyl, -C 0-6 Alkylene-C 3-8 Cycloalkyl, -C 0-6 Alkylene-3-10 membered heterocyclic group, -C 0-6 Alkylene-C 6-10 Aryl or -C 0- 6-alkylene-5-14 membered heteroaryl, wherein the above groups are optionally substituted by one or more selected from halogen, CN, NO2, OH, NH2, C 1-6 Alkyl or C 1-6 or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group;

[0193] R x Selected from H, D, OH, OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl; wherein R x Optionally deuterated, up to fully deuterated;

[0194] n=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0195] p = 0 or 1;

[0196] Preferably,

[0197] X1 is a chemical bond, O or S;

[0198] R1 is cyclohexyl, which is optionally deuterated, up to fully deuterated;

[0199] Any of R is -C(OL2)(R c )(R d ), the other two are R b ;

[0200] L1 and L2 are selected from DMTr or -P(OCH2CH2CN)(N(iPr)2);

[0201] R a 、R b 、R c and R d Independently selected from H or D.

[0202] 19. The compound of formula (X) according to technical solution 17, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, which is a compound of formula (Ia), (Ib) or (Ic):

[0203] in,

[0204] X1 is a chemical bond, O or S;

[0205] R1 is selected from C 1-20 Alkyl, C 1-20 Halogenated alkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl, 5-14 membered heteroaryl or -[C(O)] p -C 1-20 Alkylene-(OCH2CH2) n R x , which are optionally replaced by one or more radicals selected from halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl or 5-14 membered heteroaryl; wherein R1 is optionally deuterated, up to fully deuterated;

[0206] L1 and L2 are selected from H, a reactive phosphorus group or a protecting group;

[0207] R a 、R b 、R c and R d independently selected from H, D, halogen, CN, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -LC3-8 Cycloalkyl or -L-4-7 membered heterocyclic group; wherein R a 、R b 、R c and R d Optionally deuterated, up to fully deuterated;

[0208] Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated;

[0209] R', R" and R'" are independently selected from H, -C 1-6 Alkylene-OH, -C 1-6 Alkylene-NH2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2- 6 alkenyl, C 2-6 Alkynyl, -C 0-6 Alkylene-C 3-8 Cycloalkyl, -C 0-6 Alkylene-3-10 membered heterocyclic group, -C 0-6 Alkylene-C 6-10 Aryl or -C 0- 6-alkylene-5-14 membered heteroaryl, wherein the above groups are optionally substituted by one or more selected from halogen, CN, NO2, OH, NH2, C 1-6 Alkyl or C 1-6 or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group;

[0210] R x Selected from H, D, OH, OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl; wherein R x Optionally deuterated, up to fully deuterated;

[0211] n=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0212] p = 0 or 1;

[0213] Preferably,

[0214] X1 is a chemical bond, O or S;

[0215] R1 is selected from C 3-6Cycloalkyl or 5-6 membered heterocyclyl, which is optionally deuterated up to fully deuterated;

[0216] L1 and L2 are selected from DMTr or -P(OCH2CH2CN)(N(iPr)2);

[0217] R a 、R b 、R c and R d Independently selected from H, D, halogen, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0218] Preferably,

[0219] X1 is a chemical bond, O or S;

[0220] R1 is C 4-6 Cycloalkyl, which is optionally deuterated, up to fully deuterated;

[0221] L1 and L2 are selected from DMTr or -P(OCH2CH2CN)(N(iPr)2);

[0222] R a 、R b 、R c and R d Independently selected from H, D, halogen, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0223] Preferably,

[0224] X1 is a chemical bond or O;

[0225] R1 is cyclohexyl, which is optionally deuterated, up to fully deuterated;

[0226] L1 and L2 are selected from DMTr or -P(OCH2CH2CN)(N(iPr)2);

[0227] R a 、R b 、R c and R d Independently selected from H or D.

[0228] 20. The compound of formula (X) of technical solution 17, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, which is a compound of formula (II):

[0229] in,

[0230] Y is a chemical bond or C(Ra )(R b );

[0231] R is selected from H, D, -L-OR' or -L-NR"R"', which are optionally deuterated, up to fully deuterated;

[0232] L1 and L2 are selected from H, a reactive phosphorus group or a protecting group;

[0233] R a and R b independently selected from H, D, halogen, CN, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -LC 3-8 Cycloalkyl or -L-4-7 membered heterocyclic group; wherein R a and R b Optionally deuterated, up to fully deuterated;

[0234] Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated;

[0235] R', R" and R'" are independently selected from H, -C 1-6 Alkylene-OH, -C 1-6 Alkylene-NH2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2- 6 alkenyl, C 2-6 Alkynyl, -C 0-6 Alkylene-C 3-8 Cycloalkyl, -C 0-6 Alkylene-3-10 membered heterocyclic group, -C 0-6 Alkylene-C 6-10 Aryl or -C 0- 6-alkylene-5-14 membered heteroaryl, wherein the above groups are optionally substituted by one or more selected from halogen, CN, NO2, OH, NH2, C 1-6 Alkyl or C 1-6 or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group;

[0236] Preferably,

[0237] Y is a chemical bond or C(R a )(R b );

[0238] R is selected from H, D, -L-OR' or -L-NR"R"', which are optionally deuterated, up to fully deuterated;

[0239] L1 and L2 are selected from DMTr or -P(OCH2CH2CN)(N(iPr)2);

[0240] R a and R b are independently selected from H, D, halogen, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Haloalkyl, -LC 3-8 Cycloalkyl or 4-7 membered heterocyclic group; wherein R a and R b Optionally deuterated, up to fully deuterated;

[0241] Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated;

[0242] R', R" and R'" are independently selected from H, C 1-6 Alkyl or C 1-6 Haloalkyl; or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group;

[0243] Preferably,

[0244] Y is a chemical bond or C(R a )(R b );

[0245] R is selected from H, D or -C 1-6 Alkylene-NR"R"', which is optionally deuterated, up to fully deuterated;

[0246] L1 and L2 are selected from DMTr or -P(OCH2CH2CN)(N(iPr)2);

[0247] R a and R b Independently selected from H, D, halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, -C 0-6 Alkylene-NR"R"', C 4-6 Cycloalkyl or 5-6 membered heterocyclic group; wherein R a and R b Optionally deuterated, up to fully deuterated;

[0248] wherein R" and R"' are independently selected from H, C 1-4 Alkyl or C 1-4 Haloalkyl; or R", R'" and the nitrogen atom to which they are attached together form a 5-6 membered heterocyclic group;

[0249] Preferably,

[0250] Y is a chemical bond or C(R a )(R b );

[0251] R is selected from H, D, CH2NMe2 or CH2-morpholinyl, which is optionally deuterated, up to fully deuterated;

[0252] L1 and L2 are selected from DMTr or -P(OCH2CH2CN)(N(iPr)2);

[0253] R a and R b is independently selected from H, D, Me, NMe2, morpholinyl, CH2NMe2 or CH2-morpholinyl, which are optionally deuterated up to fully deuterated.

[0254] 21. The compound of any one of technical solutions 1 to 20, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the compound is selected from the following:

[0255] 22. An oligonucleotide comprising one or more compounds of formula (X') at the 5' and / or 3' end:

[0256] in,

[0257] R2 is selected from H, D or

[0258] R is selected from H, D, -L-OR', -L-NR"R"', R b or It is optionally deuterated, up to fully deuterated;

[0259] The condition is that one of R and R2 exists group, in which represents the chemical bond connecting to the nucleotides of the oligonucleotide, and the terminal Connected to H, reactive phosphorus groups or protecting groups,

[0260] The definitions of other groups are as defined in any one of technical solutions 1-21.

[0261] 23. The oligonucleotide of technical solution 22, wherein the compound of formula (X') is the following compound:

[0262] wherein each group is defined as in any one of technical solutions 1-22.

[0263] 24. The oligonucleotide of technical solution 22, wherein the compound of formula (X') is the following compound:

[0264] The order of 5'–>3' is from the to connect.

[0265] 25. A double-stranded RNA comprising a sense strand and an antisense strand with fully complementary sequences, each strand having 14 to 30 nucleotides, the nucleotides being linked by phosphate groups, phosphorothioate groups, or other linking molecules, wherein the sense strand has the following structure:

[0266] 5'(NT1) n1 -(NT2) n2 -(NT3) n3 3'

[0267] in,

[0268] NT1 and NT3 are independently compounds of formula (X');

[0269] NT2 is a modified or unmodified nucleotide;

[0270] n1 is 0, 1, 2, or 3;

[0271] n2 is an integer from 14 to 30;

[0272] n3 is 0, 1, 2, or 3;

[0273] And n1 and n3 are not 0 at the same time;

[0274] in,

[0275] R2 is selected from H, D or

[0276] R is selected from H, D, -L-OR', -L-NR"R"', R b or It is optionally deuterated, up to fully deuterated;

[0277] X1 is a chemical bond, O or S;

[0278] X2 is a chemical bond or NR1;

[0279] Y is a chemical bond or [C(R a )(R b )] 1-4 ;

[0280] R1 is selected from C 1-20Alkyl, C 1-20 Halogenated alkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl, 5-14 membered heteroaryl or -[C(O)] p -C 1-20 Alkylene-(OCH2CH2) n R x , which are optionally replaced by one or more radicals selected from halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl or 5-14 membered heteroaryl; wherein R1 is optionally deuterated, up to fully deuterated;

[0281] R a 、R b 、R c and R d independently selected from H, D, halogen, CN, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -LC 3-8 Cycloalkyl or -L-4-7 membered heterocyclic group; wherein R a 、R b 、R c and R d Optionally deuterated, up to fully deuterated;

[0282] Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated;

[0283] R', R" and R'" are independently selected from H, -C 1-6 Alkylene-OH, -C 1-6 Alkylene-NH2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2- 6 alkenyl, C 2-6 Alkynyl, -C 0-6 Alkylene-C 3-8 Cycloalkyl, -C 0-6 Alkylene-3-10 membered heterocyclic group, -C0-6 Alkylene-C 6-10 Aryl or -C 0- 6-alkylene-5-14 membered heteroaryl, wherein the above groups are optionally substituted by one or more selected from halogen, CN, NO2, OH, NH2, C 1-6 Alkyl or C 1-6 or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group;

[0284] R x Selected from H, D, OH, OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl; wherein R x Optionally deuterated, up to fully deuterated;

[0285] n=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0286] p = 0 or 1;

[0287] R s Selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0288] m = 0, 1, 2, 3, or 4;

[0289] The condition is that one of R and R2 exists group, in which represents the chemical bond connecting to the nucleotides of the oligonucleotide, and the terminal Attached to H, a reactive phosphorus group or a protecting group.

[0290] 26. The double-stranded RNA of technical solution 25, wherein X1 is a chemical bond or O, preferably O.

[0291] 27. The double-stranded RNA of technical solution 25 or 26, wherein X2 is a chemical bond.

[0292] 28. The double-stranded RNA of technical solution 25 or 26, wherein X2 is NR1.

[0293] 29. The double-stranded RNA of any one of technical solutions 25-28, wherein Y is a chemical bond.

[0294] 30. The double-stranded RNA of any one of technical solutions 25-28, wherein Y is [C(R a )(R b )] 1-4 , preferably C(R a )(R b ).

[0295] 31. The double-stranded RNA of any one of technical solutions 25-30, wherein R1 is selected from C 1-20 Alkyl, C 1-20 Halogenated alkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl, preferably selected from C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl, more preferably selected from C 3-8 Cycloalkyl or 3-10 membered heterocyclic group, especially C 5-6 Cycloalkyl.

[0296] 32. The double-stranded RNA of technical solution 31, wherein R1 is optionally replaced by one or more selected from halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl or C 1-6 The haloalkyl group is substituted, preferably by one or more groups selected from halogen, CN, OR', SR', NR"R"', C 1-6 Alkyl or C 1-6 wherein R1 is optionally substituted with deuterium, up to full deuteration.

[0297] 33. The double-stranded RNA of any one of technical solutions 25-32, wherein R2 is selected from H or D.

[0298] 34. The double-stranded RNA of any one of technical solutions 25-32, wherein R2 is

[0299] 35. The double-stranded RNA of any one of technical solutions 25-34, wherein R is selected from H, D, -L-OR', -L-NR"R"' or R b , which is optionally deuterated, up to fully deuterated.

[0300] 36. The double-stranded RNA of any one of technical solutions 25-34, wherein one of R is It is optionally deuterated, up to full deuteration.

[0301] 37. The double-stranded RNA of any one of technical solutions 25-36, wherein R a 、R b 、R c and R d Independently selected from H, D, halogen, CN, -C 0-6 Alkyl-OR', -C 0-6 Alkyl-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, preferably H, D, halogen, CN, C 1-6 Alkyl or C 1-6 Haloalkyl; wherein R a 、R b 、R c and R d Optionally deuterated, up to fully deuterated.

[0302] 38. The double-stranded RNA of any one of technical solutions 25-37, wherein the compound of formula (X') is selected from the following general formula:

[0303] Wherein each group is as defined in technical solutions 25-37.

[0304] 39. The double-stranded RNA of technical solution 38, wherein the compound of formula (X') is a compound of formula (I'):

[0305] in,

[0306] X1 is a chemical bond, O or S;

[0307] R1 is selected from C 1-20 Alkyl, C 1-20 Halogenated alkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl, 5-14 membered heteroaryl or -[C(O)] p -C 1-20 Alkylene-(OCH2CH2) n R x, which are optionally replaced by one or more radicals selected from halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl or 5-14 membered heteroaryl; wherein R1 is optionally deuterated, up to fully deuterated;

[0308] Any one of R The other two are R b ;

[0309] R a 、R b 、R c and R d independently selected from H, D, halogen, CN, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -LC 3-8 Cycloalkyl or -L-4-7 membered heterocyclic group; wherein R a 、R b 、R c and R d Optionally deuterated, up to fully deuterated;

[0310] Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated;

[0311] R', R" and R'" are independently selected from H, -C 1-6 Alkylene-OH, -C 1-6 Alkylene-NH2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2- 6 alkenyl, C 2-6 Alkynyl, -C 0-6 Alkylene-C 3-8 Cycloalkyl, -C 0-6 Alkylene-3-10 membered heterocyclic group, -C 0-6 Alkylene-C 6-10 Aryl or -C 0- 6-alkylene-5-14 membered heteroaryl, wherein the above groups are optionally substituted by one or more selected from halogen, CN, NO2, OH, NH2, C 1-6 Alkyl or C 1-6or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group;

[0312] R x Selected from H, D, OH, OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl; wherein R x Optionally deuterated, up to fully deuterated;

[0313] n=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0314] p = 0 or 1;

[0315] Preferably,

[0316] X1 is a chemical bond, O or S;

[0317] R1 is cyclohexyl, which is optionally deuterated, up to fully deuterated;

[0318] Any one of R The other two are R b ;

[0319] R a 、R b 、R c and R d Independently selected from H or D.

[0320] 40. The double-stranded RNA of technical solution 38, wherein the compound of formula (X') is a compound of formula (Ia'), (Ib') or (Ic'):

[0321] in,

[0322] X1 is a chemical bond, O or S;

[0323] R1 is selected from C 1-20 Alkyl, C 1-20 Halogenated alkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl, 5-14 membered heteroaryl or -[C(O)] p -C 1-20 Alkylene-(OCH2CH2) n R x, which are optionally replaced by one or more radicals selected from halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl or 5-14 membered heteroaryl; wherein R1 is optionally deuterated, up to fully deuterated;

[0324] R a 、R b 、R c and R d independently selected from H, D, halogen, CN, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -LC 3-8 Cycloalkyl or -L-4-7 membered heterocyclic group; wherein R a 、R b 、R c and R d Optionally deuterated, up to fully deuterated;

[0325] Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated;

[0326] R', R" and R'" are independently selected from H, -C 1-6 Alkylene-OH, -C 1-6 Alkylene-NH2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2- 6 alkenyl, C 2-6 Alkynyl, -C 0-6 Alkylene-C 3-8 Cycloalkyl, -C 0-6 Alkylene-3-10 membered heterocyclic group, -C 0-6 Alkylene-C 6-10 Aryl or -C 0- 6-alkylene-5-14 membered heteroaryl, wherein the above groups are optionally substituted by one or more selected from halogen, CN, NO2, OH, NH2, C 1-6 Alkyl or C 1-6 or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group;

[0327] R x Selected from H, D, OH, OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl; wherein R x Optionally deuterated, up to fully deuterated;

[0328] n=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0329] p = 0 or 1;

[0330] Preferably,

[0331] X1 is a chemical bond, O or S;

[0332] R1 is selected from C 3-6 Cycloalkyl or 5-6 membered heterocyclyl, which is optionally deuterated up to fully deuterated;

[0333] R a 、R b 、R c and R d Independently selected from H, D, halogen, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0334] Preferably,

[0335] X1 is a chemical bond, O or S;

[0336] R1 is C 4-6 Cycloalkyl, which is optionally deuterated, up to fully deuterated;

[0337] R a 、R b 、R c and R d Independently selected from H, D, halogen, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0338] Preferably,

[0339] X1 is a chemical bond or O;

[0340] R1 is cyclohexyl, which is optionally deuterated, up to fully deuterated;

[0341] Ra 、R b 、R c and R d Independently selected from H or D.

[0342] 41. The double-stranded RNA of technical solution 38, wherein the compound of formula (X') is a compound of formula (II'):

[0343] in,

[0344] Y is a chemical bond or C(R a )(R b );

[0345] R is selected from H, D, -L-OR' or -L-NR"R"', which are optionally deuterated, up to fully deuterated;

[0346] R a and R b independently selected from H, D, halogen, CN, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -LC 3-8 Cycloalkyl or -L-4-7 membered heterocyclic group; wherein R a and R b Optionally deuterated, up to fully deuterated;

[0347] Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated;

[0348] R', R" and R'" are independently selected from H, -C 1-6 Alkylene-OH, -C 1-6 Alkylene-NH2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2- 6 alkenyl, C 2-6 Alkynyl, -C 0-6 Alkylene-C 3-8 Cycloalkyl, -C 0-6 Alkylene-3-10 membered heterocyclic group, -C 0-6 Alkylene-C 6-10 Aryl or -C 0- 6-alkylene-5-14 membered heteroaryl, wherein the above groups are optionally substituted by one or more selected from halogen, CN, NO2, OH, NH2, C 1-6 Alkyl or C 1-6 or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group;

[0349] Preferably,

[0350] Y is a chemical bond or C(R a )(R b );

[0351] R is selected from H, D, -L-OR' or -L-NR"R"', which are optionally deuterated, up to fully deuterated;

[0352] R a and R b are independently selected from H, D, halogen, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Haloalkyl, -LC 3-8 Cycloalkyl or 4-7 membered heterocyclic group; wherein R a and R b Optionally deuterated, up to fully deuterated;

[0353] Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated;

[0354] R', R" and R'" are independently selected from H, C 1-6 Alkyl or C 1-6 Haloalkyl; or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group;

[0355] Preferably,

[0356] Y is a chemical bond or C(R a )(R b );

[0357] R is selected from H, D or -C 1-6 Alkylene-NR"R"', which is optionally deuterated, up to fully deuterated;

[0358] R a and R b Independently selected from H, D, halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, -C 0-6 Alkylene-NR"R"', C 4-6 Cycloalkyl or 5-6 membered heterocyclic group; wherein R a and R b Optionally deuterated, up to fully deuterated;

[0359] wherein R" and R"' are independently selected from H, C 1-4 Alkyl or C 1-4Haloalkyl; or R", R'" and the nitrogen atom to which they are attached together form a 5-6 membered heterocyclic group;

[0360] Preferably,

[0361] Y is a chemical bond or C(R a )(R b );

[0362] R is selected from H, D, CH2NMe2 or CH2-morpholinyl, which is optionally deuterated, up to fully deuterated;

[0363] R a and R b is independently selected from H, D, Me, NMe2, morpholinyl, CH2NMe2 or CH2-morpholinyl, which are optionally deuterated up to fully deuterated.

[0364] 42. The double-stranded RNA according to any one of technical solutions 25 to 41, wherein the compound of formula (X') is the following compound:

[0365] The order of 5'–>3' is from the to connect.

[0366] 43. The double-stranded RNA of any one of technical solutions 25-42, wherein NT1, n1, NT3 and n3 are selected from the following table, and the nucleotides are preferably linked by phosphate groups or phosphorothioate groups:

[0367] 44. The double-stranded RNA of technical solution 25, wherein n1 is 0, NT3 is selected from the compound of formula (I'), (Ia'), (Ib') or (Ic'), and n3 is 1 or 2, preferably 2.

[0368] 45. The double-stranded RNA of technical solution 25, wherein n1 is 1, NT1 is a compound of formula (II'), and n3 is 0.

[0369] 46. ​​The double-stranded RNA of technical solution 25, wherein n1 is 0, NT3 is a compound of formula (II'), and n3 is 1.

[0370] 47. The double-stranded RNA of technical solution 25, wherein n1 is 1, NT1 is selected from the compound of formula (I'), (Ia'), (Ib') or (Ic'), and n3 is 0.

[0371] 48. The double-stranded RNA of technical solution 25, wherein n1 is 1, NT1 is a compound of formula (II'), NT3 is selected from a compound of formula (I'), (Ia'), (Ib') or (Ic'), and n3 is 1 or 2, preferably 2.

[0372] 49. The double-stranded RNA of technical solution 25, wherein n1 is 2, NT1 is a compound of formula (I'), NT3 is selected from a compound of formula (I'), (Ia'), (Ib') or (Ic'), and n3 is 1 or 2, preferably 2.

[0373] 50. The double-stranded RNA of technical solution 25, wherein n1 is 2, NT1 is a compound of formula (I'), (Ia'), (Ib') or (Ic'), NT3 is selected from a compound of formula (I'), (Ia'), (Ib') or (Ic'), and n3 is 2.

[0374] 51. The double-stranded RNA of technical solution 25, wherein n1 is 1, NT1 is selected from the compound of formula (I'), (Ia'), (Ib') or (Ic'), NT3 is selected from the compound of formula (I'), (Ia'), (Ib') or (Ic'), and n3 is 1.

[0375] 52. The double-stranded RNA of technical solution 25, wherein n1 is 1, NT1 is selected from the compound of formula (I'), (Ia'), (Ib') or (Ic'), NT3 is selected from the compound of formula (I'), (Ia'), (Ib') or (Ic'), and n3 is 2.

[0376] 53. The double-stranded RNA of any one of technical solutions 25-52, which is further conjugated to a ligand portion comprising N-acetylgalactosamine, preferably conjugated to the ligand portion at its positive strand, preferably conjugated to the ligand portion at the 3' end of the positive strand, preferably conjugated to the ligand portion at the 5' end of the positive strand.

[0377] 54. The double-stranded RNA of technical solution 53, wherein the ligand comprises one or more GalNAc.

[0378] 55. The double-stranded RNA of technical solution 54, wherein a ligand comprising GalNAc is coupled to the 5' end of the sense strand, and the ligand is selected from NAG37.

[0379] 56. The double-stranded RNA of technical solution 54, wherein a ligand comprising GalNAc is coupled to the 3' end of the sense strand, and the ligand is selected from L96, GL6 or GL12, preferably GL6.

[0380] 57. The double-stranded RNA of technical solution 56, wherein n1 is 1, NT1 is a compound of formula (II'), NT3 is selected from a compound of formula (I'), (Ia'), (Ib') or (Ic'), n3 is 2, and GL6 is coupled to the 3' end of the positive chain.

[0381] 58. The double-stranded RNA of technical solution 57, wherein n1 is 1, NT1 is a compound of formula (II'), NT3 is selected from a compound of formula (I'), (Ia'), (Ib') or (Ic'), n3 is 2, and GL12 is coupled to the 3' end of the positive chain.

[0382] 59. The double-stranded RNA of any one of technical solutions 25-58, which is selected from small interfering RNA (siRNA) and short hairpin RNA (shRNA).

[0383] 60. A double-stranded RNA comprising a sense strand and an antisense strand with fully complementary sequences, each strand having 14 to 30 nucleotides, the nucleotides being linked by phosphate groups, phosphorothioate groups, or other linking molecules, wherein the sense strand has one of the following modification patterns:

[0384] Wherein N is a modified or unmodified nucleotide;

[0385] n is an integer from 14 to 30;

[0386] STM is selected from the compounds of formula (I'), (Ia'), (Ib') or (Ic');

[0387] LG is a ligand containing GalNAc, preferably GL6;

[0388] s is phosphorothioate.

[0389] 61. The double-stranded RNA of technical solution 60, wherein the modification pattern of the positive chain is selected from MP1-2, MP2-1, MP3-2, MP4-9 or MP4-10, preferably MP1-2 or MP2-1.

[0390] 62. The double-stranded RNA of technical solution 61, wherein the modification pattern of the positive strand is MP1-2.

[0391] 63. The double-stranded RNA of technical solution 61, wherein the modification pattern of the positive strand is MP2-1.

[0392] 64. The double-stranded RNA of technical solution 61, wherein the modification pattern of the positive strand is MP3-2.

[0393] 65. The double-stranded RNA of technical solution 61, wherein the modification pattern of the positive strand is MP4-9.

[0394] 66. The double-stranded RNA of technical solution 61, wherein the modification pattern of the positive strand is MP4-10.

[0395] 67. The double-stranded RNA of any one of technical solutions 61-66, wherein STM is STM1, STM2 or STM3, preferably STM1.

[0396] 68. The double-stranded RNA of any one of technical solutions 25-67, wherein the sense strand comprises one of the following nucleotide sequences:

[0397] FIN-FIN-FIN-CmsAmsGmUmGfUmUfCfUfUmGmCmUmCmUmAmUmAmAm-STM1-STM1, FIN-FIN-FIN-CmsAmsGmUmGfUmUfCfUfUmGmCmUmCmUmAmUmAmAm-STM2aP1-STM2aP1, FIN-FIN-FIN-Cms AmsGmUmGfUmUfCfUfUmGmCmUmCmUmAmUmAmAm-STM2aP2-STM2aP2, FIN-FIN-FIN-CmsAmsG mUmGfUmUfCfUfUmGmCmUmCmUmAmUmAmAm-STM3P1-STM3P1, FIN-FIN-FIN-CmsAmsGmUmGfUm UfCfUfUmGmCmUmCmUmAmUmAmAm-STM3P2-STM3P2, CmsAmsGmUmGfUmUfCfUfUmGmCmUmCmUm AmUmAmAm-STM1-STM1s-GL6, STM1s-AmsAmCmAmGmUmGfUmUfCfUfUmGmCmUmCmUmAmUmAmAms -STM1s-GL6, STM1s-UmsCmCmAmCmGmUmUmGfCfUfUmGmAmAmAmUmUmGmAmAms-STM1s-GL6, ST M1-STM1-UmsCmCmAmCmGmUmUmGfCfUfUmGmAmAmAmUmUmGmAmsAm-STM1-STM1s-GL6, STM1s- UmsCmCmAmCmGmUmUmGfCfUfUmGmAmAmAmUmUmGmAmAm-STM1-STM1s-GL6, STM1-STM1-UmsCmCmAmCmGmUmUmGfCfUfUmGmAmAmUmUmGmAmAm-STM1-STM1-GL6, ST M1s-AmsAmAmAmGmGmGmAmCfAfGfUmAmUmUmCmUmCmAmGmUms-STM1s-GL6 or STM1-STM1-AmsAmAmAmGmGmGmAmCfAfGfUmAmUmUmCmUmCmAmGmsUm-STM1-STM1s-GL6.

[0398] 69. The double-stranded RNA of technical solution 68, wherein the antisense strand comprises the following nucleotide sequence:

[0399] UmsUfsAmUmAmGfAmGmCmAmAmGmAmAfCmAfCmUmGmsUmsUm, UmsUfsCmAfAmUfUmUfCmAfAmGfCmAfAmCfGmUfGmsGfsAm, UmsAfsCmsUfGmAfUmCfAmAfAmUfAmUfGmUfUmGfAmGfsCm or AmsCfsUmGfAmGfAmAfUmAfCmUfGmUfCmCfCmUfUmsUfsUm.

[0400] 70. A vector comprising a nucleotide sequence encoding the double-stranded RNA according to any one of the aforementioned technical solutions 25-69.

[0401] 71. A cell containing the double-stranded RNA according to any one of technical solutions 25-69 or the vector according to technical solution 70.

[0402] 72. A pharmaceutical composition comprising the double-stranded RNA according to any one of technical solutions 25 to 69, the vector according to technical solution 70, or the cell according to technical solution 71, and optionally a pharmaceutically acceptable carrier or excipient.

[0403] 73. A kit comprising the double-stranded RNA according to any one of technical solutions 25-69, the vector according to technical solution 70, or the cell according to technical solution 71. Example

[0404] The single-stranded sequences used in the examples are as follows:

[0405] The double-stranded sequences used in the examples are as follows:

[0406] In this article, the meanings of the abbreviations are as follows:

[0407] A, U, G and C represent the natural ribonucleotides adenine, uracil, guanine and cytosine, respectively.

[0408] T represents thymidine ribonucleotide.

[0409] d indicates that the nucleotide adjacent to its right is a deoxyribonucleotide. For example, dA, dT, dG, and dC represent adenine deoxyribonucleotide, thymine deoxyribonucleotide, guanine deoxyribonucleotide, and cytosine deoxyribonucleotide, respectively.

[0410] i represents inosine ribonucleotide.

[0411] m indicates that the nucleotide adjacent to its left is a 2'-OCH3 modified nucleotide. For example, Am, Um, Gm, and Cm represent 2'-OCH3 modified A, U, G, and C.

[0412] f indicates that the nucleotide adjacent to its left is a 2'-F modified nucleotide. For example, Af, Uf, Gf, and Cf represent 2'-F modified A, U, G, and C, respectively.

[0413] "s" indicates that the two adjacent nucleotides and / or delivery vectors are linked by phosphorothioate.

[0414] VP indicates that the adjacent nucleotide on the right is a vinyl phosphate-modified nucleotide.

[0415] IB represents an inverted abasic deoxyribonucleotide, which can include the following three structures depending on its position / linking method in siRNA.

[0416] The numbering and structure of the compounds of the present invention in oligonucleotides are as follows, wherein the sequence of 5'-->3' from the compound to connect.

[0417] Specifically, according to the order of 5'–>3', if the corresponding structure is located in the middle of the nucleic acid chain, means that it is connected to the 3' carbon or corresponding position of the previous nucleotide or nucleotide analog through a phosphate group, a phosphorothioate group or other linking group, Indicates connection to the 5' carbon or corresponding position of the next nucleotide or nucleotide analog via a phosphate group, a phosphorothioate group or other linking group; if the corresponding structure is at the terminal position of the nucleic acid chain, it may also indicate connection to hydrogen, a terminal modification, a terminal protecting group, a delivery vector or other structure that can be used for the terminal of the nucleic acid chain; the optical isomers P1 / P2 are consistent with the corresponding P1 / P2 in the examples:

[0418] L96 represents a GalNAc delivery vector of the following structure well known in the art, wherein Indicates the position of the siRNA attached via a phosphate or phosphorothioate group:

[0419] NAG37 represents a GalNAc delivery vector of the following structure well known in the art, wherein Indicates the location where the siRNA is attached:

[0420] GL6 represents a GalNAc delivery vector of the following structure, wherein Indicates the position of the siRNA attached via a phosphate or phosphorothioate group:

[0421] GL12 represents a GalNAc delivery vector of the following structure, wherein Indicates the position of the siRNA attached via a phosphate or phosphorothioate group:

[0422] FIN-FIN-FIN- represents a GalNAc delivery vector of the following structure, wherein Indicates the location where the siRNA is attached:

[0423] Example 1: Synthesis of Compounds E1-P1 and E1-P2

[0424] 1. Preparation of Compound 1c

[0425] To a mixture of compounds 1a (29.0 g, 188 mmol) and 1b (22.2 g, 226 mmol) in dichloromethane (580 mL) was added acetic acid (2.16 mL, 37.3 mmol). After stirring at room temperature for 3 hours, sodium acetate borohydride (80.0 g, 377 mmol) was added and stirring continued at room temperature for 12 hours. After concentration under reduced pressure, the residue was separated by silica gel column chromatography (dichloromethane / methanol = 10:1) to obtain the title compound 1c (58.9 g, 97.6% yield).

[0426] 1 H NMR (400 MHz CDCl3)δ9.50(brs,3H),4.07-3.94(m,3H),3.64(d,J=4.8Hz,2H),3.39-3.36( m,2H),3.30-3.27(m,2H),2.88(tt,J1=11.6Hz,J2=3.2Hz,1H),2.74(td,J1=11 .6Hz,J2=4.8Hz,1H),2.64(t,J=11.2Hz,1H),2.08-2.05(m,2H),2.01(s,6.54H ),1.89-1.86(m,2H),1.69-1.65(m,1H),1.44-1.23(m,4H),1.17-1.06(m,1H).

[0427] 2. Preparation of Compound 1d

[0428] Oxalyl chloride (4.37 g, 34.4 mmol) was dissolved in dichloromethane (100 mL), evacuated, and filled with nitrogen. The temperature was cooled to -65°C, and dimethyl sulfoxide (2.69 g, 34.4 mmol) was added dropwise. The reaction was stirred for 15 minutes, followed by the addition of compound 1c (5.0 g, 15.6 mmol) and triethylamine (16.3 mL, 117 mmol), and stirring continued for 1 hour. The reaction was quenched with water. After the reaction solution was warmed to room temperature, it was extracted with ethyl acetate (200 mL x 2). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the organic solvent was concentrated under reduced pressure to afford the title compound 1d (12.0 g), which was used directly in the next step.

[0429] 1 H NMR(400MHz CDCl3)δ9.65(s,1H),4.02-3.96(m,2H),3.75-3.69(m,3H),2.97-2.94(m,1H),2.70-2.67(m,1H),2.46-2 .40(m,1H),2.37-2.31(1H),2.24-2.22(m,1H),1.85-1.78(m,4H),1.63-1.60(m,1H),1.28-1.05(m,5H).

[0430] 3. Preparation of Compound 1e

[0431] Triphenylmethylphosphonium bromide (16.3 g, 45.6 mmol) was suspended in tetrahydrofuran (150 mL), evacuated, and filled with nitrogen. Potassium tert-butoxide (45.6 mL, 1 M solution in tetrahydrofuran) was added, and the mixture was stirred at room temperature for 1 hour. Compound 1d (3.00 g, 15.2 mmol) was dissolved in tetrahydrofuran (10 mL) and added dropwise to the mixture. The reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was poured into saturated ammonium chloride solution, adjusted to pH 8 with saturated sodium bicarbonate, and extracted with ethyl acetate (100 mL x 2). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and filtered. The organic solvent was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1) to obtain the title compound 1e (10.1 g, 59.5% yield).

[0432] 1H NMR(400MHz CDCl3)δ5-85-5.77(m,1H),5.33-5.28(m,1H),5.17-5.15(m,1H),4.04-3 .97(m,1H),3.94-3.91(m,1H),3.72-3.67(m,1H),2.82-2.79(m,1H),2.72 -2.70(m,1H),2.39-2.33(m,1H),2.26-2.17(m,1H),2.13-2.07(m,1H),1. 89-1.87(m,2H),1.81-1.78(m,2H),1.64-1.62(m,1H),1.26-1.09(m,5H).

[0433] 4. Preparation of Compound 1f

[0434] To a solution of compound 1e (6.00 g, 21.5 mmol) and N-methylmorpholine oxide (3.28 g, 27.9 mmol) in acetone was added an aqueous solution of K₂OsO₄.2H₂O (158 mg, 430 μmol). The reaction was stirred at room temperature for 12 hours. The reaction mixture was filtered, the filtrate was concentrated under reduced pressure, and the organic solvent was concentrated. The residue was then separated by silica gel column chromatography (dichloromethane / methanol = 10:1) to afford the title compound 1f (9.01 g, 82.1% yield).

[0435] 1 H NMR(400MHz CD3OD)δ3.91-3.84(m,1H),3.69-3.44(m,5H),3.10-2.74(m,2H),2.38-2.11(m, 3H),1.96-1.94(m,2H),1.84-1.81(m,2H),1.67-1.64(m,1H),1.34-1.10(m,5H).

[0436] 5. Preparation of Compound 1g

[0437] To a solution of compound 1f (2.01 g, 7.85 mmol) in pyridine (12 mL) was added DMTrCl (2.79 g, 8.24 mmol) and the reaction was stirred at room temperature for 1 hour. The reaction was quenched with methanol. Multiple batches were combined and concentrated under reduced pressure. The residue was separated by silica gel column chromatography (dichloromethane / methanol = 10:1) to obtain the title compound 1g (4.01 g, 77.5% yield).

[0438] 1H NMR(400MHz DMSO-d6)δ7.42-7.39(m,2H),7.31-7.18(m,7H),6.89-6.86(m,4H),4.92-4.73(m,1H),3.77-3.67(m ,7H),3.56-3.33(m,3H),3.08-2.55(m,4H),2.21-1.97(m,3H),1.78-1.54(m,5H),1.23-1.02(m,5H).

[0439] 6. Preparation of Compounds 1g-P1 and 1g-P2

[0440] Compound 1g (6.02 g, 11.2 mmol) was separated by chiral column SFC (column: DAICEL CHIRALPAK IG (250 mm*50 mm, 10 um); mobile phase: A: CO2, B: [IPA containing 0.1% NH3.H2O]; B%: 60%-60%, 13 min) to give compound 1g-P1 (3.20 g) and compound 1g-P2 (1.80 g).

[0441] Compound 1g-P1:

[0442] m / z:ES+[M+H]+532.2

[0443] HPLC: retention time 1.580 min (column: Chiralpak IG-3, 50×4.6 mm ID, 3 μm; mobile phase: A: CO 2 , B: IPA (0.1% IPA, v / v); gradient: 0-0.2 min 5% B, 0.2-1.2 min 5-50% B, 1.2-2.2 min 50% B, 2.2-2.6 min 50%-5% B, 2.6-3.0 min 5% B; flow rate: 3.4 mL / min; column temperature: 35° C.)

[0444] 1H NMR(400MHz DMSO-d6)δ7.42-7.40(m,2H),7.31-7.12(m,7H),6.88-6.86(m,4H),4.94 (d,J=6.0Hz,1H),3.73(s,6H),3.69-3.67(m,1H),3.56-3.50(m,1H),3.39 -3.37(m,1H),3.01-2.95(m,2H),2.87-2.84(m,1H),2.58-2.55(m,1H),2. 20-2.15(m,2H),2.02-1.97(m,1H),1.78-1.54(m,5H),1.23-1.01(m,5H).

[0445] Compound 1g-P2:

[0446] m / z:ES+[M+H]+532.2

[0447] HPLC: retention time 2.048 min (column: Chiralpak IG-3, 50×4.6 mm ID, 3 μm; mobile phase: A: CO 2 , B: IPA (0.1% IPA, v / v); gradient: 0-0.2 min 5% B, 0.2-1.2 min 5-50% B, 1.2-2.2 min 50% B, 2.2-2.6 min 50%-5% B, 2.6-3.0 min 5% B; flow rate: 3.4 mL / min; column temperature: 35° C.)

[0448] 1 H NMR(400MHz DMSO-d6)δ7.41-7.39(m,2H),7.31-7.15(m,7H),6.88-6.86(m,4H),4.75(d,J=4.4Hz,1H),3.85-3.65(m,7H),3.54-3.41( m,3H),3.08-3.05(m,1H),2.85-2.82(m,1H),2.59-2.57(m,2H),2.17-2.07(m,3H),1.71-1.53(m,4H),1.24-1.02(m,5H).

[0449] 7. Preparation of Compounds E1-P1

[0450] To a solution of compound 1g-P1 (1.50 g, 2.82 mmol) in dichloromethane (15 mL) were added compound 1h (1.28 g, 4.23 mmol) and DCI (366 mg, 3.10 mmol). The reaction was stirred at 0°C for 0.5 hours. After concentration under reduced pressure, the residue was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1, 0.1% TEA) to obtain the title compound E1-P1 (700 mg, 28.2% yield).

[0451] m / z:ES+[M+H]+732.5

[0452] 1 H NMR(400MHz DMSO-d6)δ7.42-7.39(m,2H),7.32-7.19(m,7H),6.89-6.85(m,4H),3.92-3.38(m,14H),3.16-2.99(m,2H),2.88-2.80(m ,1H),2.76-2.73(m,1H),2.61-2.54(m,2H),2.22-2.10(m,2H),2.05-1.98(m,1H),1.71-1.53(m,5H),1.16-0.98(m,17H).

[0453] 8. Preparation of Compounds E1-P2

[0454] To a solution of compound 1g-P2 (1.10 g, 2.07 mmol) in dichloromethane (15 mL) were added compound 1h (935 mg, 3.10 mmol) and DCI (268 mg, 2.28 mmol). The reaction was stirred at 0°C for 0.5 hours. After concentration under reduced pressure, the residue was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1, 0.1% TEA) to obtain the title compound E1-P2 (515 mg, 26.7% yield).

[0455] m / z:ES+[M+H]+732.5

[0456] 1H NMR(400MHz DMSO-d6)δ7.41-4.39(m,2H),7.32-7.20(m,7H),6.89-6.85(m,4H),3.98-3.88(m,1H),3.83-3.37(m,13H),3.26-3.07(m,1H),3. 02-2.96(m,1H),2.78-2.75(m,1H),2.70-2.55(m,3H),2.19-1.95(m,3H),1.71-1.61(m,4H),155-152(m,1H),1.16-0.97(m,17H).

[0457] Example 2: Synthesis of Compound E2

[0458] 1. Preparation of Compound 2b

[0459] At room temperature, compound 2a (38.0 g, 231 mmol), TsNHBoc (75.4 g, 278 mmol), K2CO3 (6.40 g, 46.3 mmol), and TEBA (benzyltriethylammonium bromide, 5.27 g, 23.1 mmol) were added to a three-necked flask. The reaction was allowed to proceed at 95°C for 2 hours, then cooled to 25°C. TLC (PE / EA = 2 / 1, UV 254 nm) indicated the reaction was complete. The reaction solution was diluted with water (500 mL) and extracted with dichloromethane (200 mL x 3). The organic phase was concentrated under reduced pressure, and the crude product was purified by column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the title compound 2b (40.0 g).

[0460] 1 H NMR (400MHz, CDCl3) δ7.72 (d, J = 8.4Hz, 2H), 7.28-7.39 (m, 7H), 4.72-4.86 (m, 2H) ,4.42-4.56(m,2H),3.52-3.61m,2H),3.14-3.39(m,2H),2.43(s,3H),1.47(s,9H)

[0461] 2. Preparation of Compound 2d

[0462] Compound 2b (79.0 g, 181 mmol), compound 2c (24.7 g, 150.8 mmol), potassium carbonate (4.21 g, 30.5 mmol), and TEBA (3.47 g, 15.2 mmol) were reacted at 95°C for 3 hours. TLC (PE / EA = 2 / 1, UV 254 nm) indicated the reaction was complete. The reaction solution was diluted with water (500 mL) and extracted with dichloromethane (200 mL x 3). The organic phase was concentrated under reduced pressure, and the crude product was purified by column chromatography (PE / EA = 10 / 1 to 3 / 1) to obtain the title compound 2d (40.0 g, 43.8% yield).

[0463] 1H NMR (400MHz, CDCl3) δ7.64-7.77(m,2H),7.27-7.38(m,13H),5.05-5.16(m,1H),4.50-4.59(m,4H),4.03-4.11 (m,1H),3.62-3.76(m,2H),3.44-3.58(m,3H),3.25-3.33(m,2H),3.15-3.20(m,1H),2.42(s,3H),1.46(s,9H)

[0464] 3. Preparation of Compound 2e

[0465] Compound 2d (77.0 g, 128 mmol) and triethylamine (28.6 mL, 205 mmol) were added sequentially to dichloromethane (800 mL) in an ice-water bath, followed by the slow dropwise addition of methylsulfonyl chloride (24.2 g, 212 mmol). The reaction was allowed to proceed at 25°C for 2 hours, with LCMS indicating completion. The reaction mixture was washed with water (800 mL), and the organic phase was concentrated under reduced pressure to afford the crude title compound 2e (90.0 g).

[0466] m / z: ES+[M+Na]+700.1

[0467] 1 H NMR (400MHz, CDCl3) δ7.68 (d, J = 8.4Hz, 2H), 7.28-7.39 (m, 12H), 5.10-5.20 (m, 1H), 4.93-5.02 (m, 1H), 4.52-4.61 (m, 4 H),3.72-3.88(m,2H),3.52-3.67(m,4H),3.27-3.36(m,1H),3.15-3.22(m,1H),3.04(s,3H),2.43(s,3H),1.43(s,9H).

[0468] 4. Preparation of Compound 2f

[0469] Compound 2e (90.0 g, 133 mmol) and potassium carbonate (91.8 g, 664 mmol) were added to methanol (900 mL) at room temperature. The reaction was allowed to proceed at 66°C for 2 hours. TLC (PE / EA = 2 / 1, UV 254 nm) showed the formation of new spots. The organic phase was concentrated under reduced pressure, the reaction mixture was diluted with water (200 mL), and extracted with dichloromethane (200 mL x 3). The organic phase was concentrated under reduced pressure, and the crude product was purified by column chromatography (PE / EA = 30 / 1-2 / 1) to obtain the title compound 2f (64.0 g, 99.9% yield).

[0470] 1 H NMR (400MHz, CDCl3) δ7.62 (d, J = 8.4Hz, 2H), 7.27-7.39 (m, 12H), 4.48-4.62 ( m,4H),3.93-4.08(m,2H),3.55-3.69(m,4H),2.84-3.10(m,4H),2.44(s,3H)

[0471] 5. Preparation of Compound 2g

[0472] Compound 2f (69.0 g, 143 mmol) and magnesium turnings (54.7 g, 2.28 mol) were added to methanol (400 mL) at room temperature and reacted at 66°C for 1 hour. TLC (DCM / MeOH = 10 / 1, UV 254 nm) showed complete reaction of the starting material with the formation of new spots. The reaction mixture was diluted with water (3000 mL) and saturated aqueous ammonium chloride (3000 mL), extracted with dichloromethane (1000 mL x 3), and the organic phase was washed with saturated sodium bicarbonate (300 mL x 3). The organic phase was concentrated under reduced pressure to yield 2 g (32.0 g) of the crude title compound.

[0473] 1 H NMR (400MHz, CDCl3) δ7.27-7.40(m,10H),4.57(s,4H),3.90-4.00(m,2H),3.59-3.69(m,4H),2.77-3.04(m,4H)

[0474] 6. Preparation of Compound 2h

[0475] Compound 2g (3.00 g, 9.16 mmol), compound 1b (1.90 mL, 18.3 mmol), and acetic acid (1.01 mL, 18.3 mmol) were added to methanol (30 mL) at room temperature. The reaction was allowed to proceed at 25°C for 18 hours. Sodium cyanoborohydride (2.30 g, 36.7 mmol) was then added and the mixture was allowed to react at 50°C for 4 hours. TLC (DCM / MeOH = 10 / 1) showed the formation of new spots. The reaction solution was diluted with water (50 mL) and extracted with dichloromethane (30 mL x 3). The organic phase was concentrated under reduced pressure. The crude product was purified by column chromatography (dichloromethane / methanol = 99 / 1 to 5 / 1) to afford the title compound 2h (3.00 g, 79.9% yield).

[0476] 1 H NMR (400MHz, CDCl3) δ7.28-7.41(m,10H),4.51-4.71(m,4H),4.00-4.35(m,2H),3.49- 3.77(m,4H),2.69-2.98(m,2H),1.63-2.12(m,7H),1.16-1.44(m,6H)

[0477] 7. Preparation of Compound 2i

[0478] Compound 2h (3.00 g, 7.32 mmol) was added to concentrated hydrochloric acid (10 mL, 12 M) at room temperature and reacted at 50°C for 18 hours. TLC (DCM / MeOH = 10 / 1) showed the formation of new spots. The reaction solution was concentrated under reduced pressure to give the crude title compound 2i (2.00 g).

[0479] 1 H NMR (400MHz, CD3OD) δ4.27-4.38(m,1H),4.11-4.20(m,1H),3.96-4.04(m,2H),3.60-3.68(m,3H),3.36-3.43(m,1H),3.19-3.29(m, 2H),3.11(s,1H),2.02-2.17(m,2H),1.90-2.00(m,2H),1.69-1.78(m,1H),1.57-1.69(m,1H),1.35-1.52(m,3H),1.20-1.31(m,1H)

[0480] 8. Preparation of Compound 2j

[0481] Compound 2i (2.00 g, 8.72 mmol) was dissolved in pyridine (40 mL) at room temperature, followed by the addition of DMTrCl (2.96 g, 8.72 mmol). The reaction was allowed to proceed at 25°C for 18 hours. TLC (PE / EA = 1 / 1, UV 254 nm) indicated the reaction was complete. The reaction solution was concentrated under reduced pressure, added with saturated aqueous ammonium chloride (200 mL), and extracted with DCM (100 mL x 3). The organic phase was concentrated under reduced pressure, and the crude product was purified by column chromatography (dichloromethane / methanol = 99 / 1-10 / 1) to afford the title compound 2j (1.40 g, 30.2% yield).

[0482] 9. Preparation of Compound E2

[0483] Compound 2j (660 mg, 1.24 mmol), compound 1h (374 mg, 1.24 mmol), and DCI (73.3 mg, 0.621 mmol) were added to DCM (10 mL) and reacted at 25°C for 1 hour. TLC (PE / EA = 5 / 1, PMA) indicated complete reaction. Saturated sodium bicarbonate (50 mL) was added, and the mixture was extracted with DCM (30 mL x 3). The organic phase was concentrated under reduced pressure, and the crude product was separated by column chromatography (PE / EA = 99 / 1-30 / 1) to afford the title compound E2 (350 mg, 19.3% yield).

[0484] m / z:ES+[M+H]+732.2

[0485] 1 H NMR (400MHz, CDCl3) δ7.40-7.50(m,2H),7.28-7.37(m,6H),7.21(d,J=7. 2Hz,1H),6.77-6.89(m,4H),3.67-4.07(m,11H),3.56-3.66(m,2H),3.04- 3.33(m,2H),2.30-2.96(m,6H),2.10-2.28(m,1H),1.63-1.88(m,4H),1. 47-1.52(m,1H),1.24-1.31(m,3H),1.15-1.23(m,12H),1.11-1.14(m,1H)

[0486] Example 3: Synthesis of Compounds E3-P1 and E3-P2

[0487] 1. Preparation of Compound 3a

[0488] To a solution of compound 1d (12.1 g, 61.3 mmol) in methanol (85 mL) were added potassium carbonate (42.3 g, 306.0 mmol) and paraformaldehyde (8.2 g). The mixture was heated to 70°C and stirred for 12 hours. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (dichloromethane / methanol = 10:1) to provide compound 3a (6.70 g, 29.2% yield).

[0489] 1 H NMR(400MHz CDCl3)δ4.02-3.99(m,2H),3.66(s,4H),2.72(s,2H),2.61-2.59(m,2H),2.28(br,1H),1.82-1.78(m,4H),1.64-1.61(m,1H),1.28-1.09(m,5H)

[0490] m / z: ES+[M+H]+230.1

[0491] 2. Preparation of Compound 3b

[0492] To compound 3a (500 mg, 2.18 mmol) were added pyridine (1.72 g, 21.80 mmol) and DMTrCl (775 mg, 2.29 mmol), and the reaction was stirred at room temperature for 1 hour. The reaction was quenched with methanol, and multiple batches were combined and concentrated under reduced pressure. The residue was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1) to obtain compound 3b (2.20 g, 30.4% yield).

[0493] 1 H NMR (400 MHz CDCl3)δ7.43(d,J=7.2Hz,2H),7.33-7.27(m,5H),7.23-7.19(m,1H),6.83(d, J=8.4Hz,4H),4.03-3.94(m,3H),3.80-3.76(m,7H),3.69-3.65(m,1H),3.21(d ,J=8.8Hz,1H),3.05(d,J=8.4Hz,1H),2.81(d,J=11.6Hz,1H),2.56-2.44(m,3 H),2.27-2.23(m,1H),1.80-1.78(m,4H),1.65-1.61(m,1H),1.26-1.09(m,5H)

[0494] 3. Preparation of Compounds 3b-P1 and 3b-P2

[0495] (2-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-4-cyclohexylmorpholin-2-yl)methanol (4.10 g, 7.71 mmol) was separated using chiral column SFC (column: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 um); mobile phase: [0.1% NH3H2O ​​IPA]; B%: 50%-50%, 7 min) to give 3b-P1 (2.00 g) and 3b-P2 (1.90 g).

[0496] 3b-P1 spectrum:

[0497] 1 H NMR(400MHz CDCl3)δ7.44-7.42(m,2H),7.33-7.27(m,5H),7.27-7.21(m,1H),6.84(d,J =8.8Hz,4H),4.03-3.95(m,2H),3.79(s,6H),3.76-3.67(m,1H),3.22(d,J=8 .4Hz,1H),3.05(d,J=8.8Hz,1H),2.80(d,J=11.6Hz,1H),2.56-2.47(m,3H), 2.25-2.27(m,1H),1.81-1.78(m,4H),1.65-1.61(m,1H),1.26-1.12(m,5H).

[0498] HPLC: retention time 1.389 min (Chiralpak AD-3, 50×4.6 mm ID, 3 μm; mobile phase: A: CO2, B: IPA (0.1% IPA, v / v); gradient: 0-0.2 min 5% B, 0.2-1.2 min 5-50% B, 1.2-2.2 min 50% B, 2.2-2.6 min 50%-5% B, 2.6-3.0 min 5% B; flow rate: 3.4 mL / min; column temperature: 35°C)

[0499] 3b-P2 spectrum:

[0500] 1H NMR(400MHz CDCl3)δ7.44-7.42(m,2H),7.33-7.27(m,5H),7.27-7.21(m,1H),6.83(d,J =8.8Hz,4H),4.03-3.93(m,3H),3.79(s,6H),3.76-3.68(m,1H),3.22(d,J=8 .4Hz,1H),3.06(d,J=8.8Hz,1H),2.81(d,J=11.2Hz,1H),2.56-2.47(m,3H), 2.25-2.27(m,1H),1.81-1.78(m,4H),1.65-1.62(m,1H),1.26-1.12(m,5H).

[0501] HPLC: retention time 1.559 min (Chiralpak AD-3, 50×4.6 mm ID, 3 μm; mobile phase: A: CO2, B: IPA (0.1% IPA, v / v); gradient: 0-0.2 min 5% B, 0.2-1.2 min 5-50% B, 1.2-2.2 min 50% B, 2.2-2.6 min 50%-5% B, 2.6-3.0 min 5% B; flow rate: 3.4 mL / min; column temperature: 35°C)

[0502] 4. Preparation of Compound E3-P1

[0503] To a solution of compound 3b-P1 (1.60 g, 3.01 mmol) in dichloromethane (16 mL) were added compound 1h (1.36 g, 4.51 mmol) and DCI (390 mg, 3.31 mmol). The reaction was stirred at 0°C for 0.5 hours. After concentration under reduced pressure, the residue was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1, 0.1% TEA) to obtain compound E3-P1 (1.00 g, 43.8% yield).

[0504] 1 H NMR(400MHz DMSO-d6)δ7.41-7.38(m,2H),7.31-7.19(m,7H),6.87(d,J=8.4Hz,4H),4.01-3.89(m,1H),3.73(s,6H),3.68-3.43(m,7H),3.31-3 .16(m,1H),2.98(t,J=9.6Hz,1H),2.72-2.63(m,2H),2.45-2.29(m,4H),2.10-2.07(m,1H),1.63-1.50(m,5H),1.13-1.04(m,18H).

[0505] 5. Preparation of Compound E3-P2

[0506] To a solution of compound 3b-P2 (1.50 g, 2.82 mmol) in dichloromethane (15 mL) were added compound 1h (1.28 g, 4.23 mmol) and DCI (366 mg, 3.10 mmol). The reaction was stirred at 0°C for 0.5 hours. After concentration under reduced pressure, the residue was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1, 0.1% TEA) to obtain compound E3-P2 (1.00 g, 47.8% yield).

[0507] 1 H NMR(400MHz DMSO-d6)δ7.42-7.39(m,2H),7.30-7.18(m,7H),6.86(d,J=8.4Hz,4H),4.03-3.91(m,1H),3.72-3.44(m,13H),3.32-3.17(m ,1H),2.99(t,J=9.6Hz,1H),2.72-2.63(m,2H),2.46-2.30(m,4H),2.09-2.06(m,1H),1.63-1.49(m,5H),1.13-1.04(m,18H).

[0508] Example 4: Synthesis of siRNA

[0509] The siRNA of the present invention was prepared using the solid phase phosphoramidite method known in the art. Specific methods can be found in, for example, PCT Publication Nos. WO2016081444 and WO2019105419 and are briefly described below.

[0510] 1. Preparation of siRNA with no ligand attached to the 3' end of the sense strand

[0511] 1.1 Synthesis of the positive sense chain (SS chain)

[0512] Using solid-phase phosphoramidite synthesis, a blank CPG solid support is used as the starting cycle, and nucleoside monomers are linked one by one in the 3'-5' direction according to the order of the positive strand nucleotides. Each nucleoside monomer linking involves four steps: deprotection, coupling, capping, and oxidation or thiolation, resulting in a 5 μmol oligonucleotide. The synthesis conditions are as follows:

[0513] The nucleoside monomer was provided in a 0.05 mol / L acetonitrile solution. The reaction conditions for each step were the same, i.e., the temperature was 25 degrees Celsius. Deprotection was performed three times using a 3% trichloroacetic acid-dichloromethane solution. The coupling reaction was performed twice using a 0.25 mol / L 5-ethylthiotetrazolium (ETT)-acetonitrile solution as the activator. Capping was performed twice using 10% acetic anhydride-acetonitrile and pyridine / N-methylimidazole / acetonitrile (10:14:76, v / v / v). Oxidation was performed twice using 0.05 mol / L iodine in tetrahydrofuran / pyridine / water (70 / 20 / 10, v / v / v). Thiolysis was performed twice using 0.2 mol / L phenylacetyl disulfide (PADS) in acetonitrile / 3-methylpyridine (1 / 1, v / v).

[0514] 1.2 Synthesis of antisense strand (AS strand)

[0515] Using solid-phase phosphoramidite synthesis, a blank CPG solid support is used as the starting cycle. Nucleoside monomers are linked one by one in the 3'-5' direction according to the nucleotide arrangement of the antisense strand. Each ligation step involves four steps: deprotection, coupling, capping, and oxidation or thiolation. The synthesis conditions for a 5 μmol oligonucleotide of the antisense strand are the same as those for the sense strand.

[0516] 1.3 Oligonucleotide purification and annealing

[0517] 1.3.1 Ammonolysis

[0518] Add the synthesized solid phase carrier (sense chain or antisense chain) to a 5 mL centrifuge tube, add 3% diethylamine / ammonia water (v / v), react in a constant temperature water bath at 35 degrees (or 55 degrees) for 16 hours (or 8 hours), filter, wash the solid phase carrier three times with ethanol / water, each time 1 mL, and purify the crude product after centrifugation and concentration of the filtrate.

[0519] 1.3.2 Purification

[0520] Purification and desalination methods are well known in the art. For example, a column packed with a strong anion filler can be used, and a sodium chloride-sodium hydroxide system can be used for elution and purification, and the product can be collected and stored in a tube. A gel-filled purification column can be used for desalination, and the elution system can be pure water.

[0521] 1.3.3 Annealing

[0522] According to the instructions, the sense chain (SS chain) and the antisense chain (AS chain) were mixed at a molar ratio of (SS chain / AS chain = 1 / 1.05), heated in a water bath to 70-95 degrees, maintained for 3-5 minutes, cooled naturally to room temperature, and the system was lyophilized to obtain the product.

[0523] Example 5: In vivo activity

[0524] C57BL / 6 mice (male, 18-21 g, 6-8 weeks) were randomly divided into groups. The dosage for each animal was calculated based on body weight, and the siRNA conjugate was administered as a single subcutaneous injection in a 1 mg / mL solution (0.9% sodium chloride aqueous solution as the solvent). Specifically, before the experiment, the siRNA conjugate was dissolved in 0.9% sodium chloride aqueous solution and the volume was adjusted to the desired concentration and volume. The administration volume of physiological saline (control group) and siRNA conjugate was 5 mL / kg.

[0525] The test compound was administered subcutaneously at D0; 1 mg / kg, single dose, see Table 1 for details. Table 1 Administration details for in vivo activity screening

[0526] The animals were killed on days 14, 31, and 56 after administration (i.e., D14, D31, and D56), and 10 mg of liver was collected and placed in RNAlater solution and frozen at -80°C. RNA was then extracted from the liver tissue and the target gene (mTTR) was detected by QPCR.

[0527] The QPCR detection method is known in the art, and the primer sequences used are shown in Table 2. For example, according to the operating protocol of the high-throughput tissue RNA extraction kit (Fanzhi Medical, FG0412), a nucleic acid extractor (Hangzhou Aosheng, Auto-pure96) was used to extract cell RNA; refer to PrimeScript TM II 1st Strand cDNA Synthesis Kit (Takara, 6210B) for reverse transcription; reference TaqMan TM 20 μL of Fast Advanced Master Mix (ABI, 4444965) was used for fluorescence quantitative PCR reaction (ABI, QuantStudio3).

[0528] Table 2 QPCR primer sequences

[0529] Data statistics and analysis

[0530] Calculation 2 -△△Ct The values ​​were converted into percentages to obtain the residual inhibition rate;

[0531] △△Ct=[(target gene of Ct experimental group-internal reference of Ct experimental group)-(target gene of Ct control group-internal reference of Ct control group)].

[0532] The target gene is mTTR and the internal reference is mGAPDH

[0533] The results are shown in Table 3. The experimental results show that the compounds of the present invention (DR002346, DR002349, DR002350, DR002351 and DR002352) achieved better target gene inhibition effects than the known modifications (DR002354 and DR002355).

[0534] Table 3 Results of in vivo activity screening

[0535] Example 6: Validation of the compounds of the present invention in a C57BL / 6 mouse model

[0536] With reference to the experimental steps of Example 5, the long-lasting efficacy of the test compound (see Table 4) in the C57BL / 6 mouse model was verified.

[0537] The dosage of each animal was calculated based on body weight and administered subcutaneously as a single dose. The siRNA conjugate was administered as a 1 mg / mL solution (0.9% sodium chloride aqueous solution as the solvent) at a dose of 1 mpk. Blood was collected from the orbits at various time points and serum mTTR protein was detected using an ELISA kit (Abcam, ab282297). The experimental results are shown in Table 4.

[0538] Table 4 Experimental results of compounds in C57BL / 6 mouse model

[0539] Example 7: Validation of the compounds of the present invention in a mouse HDI model

[0540] HDI animal model

[0541] Six to eight-week-old female Balb / c mice were transfected in vivo using a dual-gene stable transfection system using a tail vein high-pressure injection method. Piggy-Bac transposon plasmids (purchased from Suzhou Bangye) containing target gene cDNA sequences (Genbank accession number NM_014495.2) and Piggy-Bac helper plasmids (purchased from Suzhou Bangye) (mass ratio 1:1, total plasmid 100 μg) were injected into the mouse through a 27-gauge needle within 5-7 seconds. The delivery solution (total volume of 10% animal body weight, Mirusbio-MIR 5240) was injected into mice and placed back in their cages for observation for 30 minutes after injection. The day of modeling was designated as day 0, and serum was obtained at various time points after modeling (days 7-35) to detect SEAP expression levels.

[0542] The dual-gene stable transfection system includes a Piggy-Bac helper plasmid and a Piggy-Bac transposon plasmid. The Piggy-Bac helper plasmid provides the Piggy-Bac transposase; the Piggy-Bac transposon plasmid is based on the Piggy-Bac transposon and contains a dual-gene expression element, which contains the secreted alkaline phosphatase gene (SEAP) and the target gene (ANGPTL3).

[0543] Detection of SEAP expression

[0544] Kit (Phospha-Light TM SEAP reporter gene assay system, Invitrogen, T1016) standard was diluted 2-fold with an initial concentration of 15 mU / mL to obtain 7 concentration points.

[0545] The CSPD substrate was mixed with the reaction buffer at a ratio of 1:20 to prepare the reaction solution. 5× dilution buffer was diluted to 1× dilution buffer with DNase- and RNase-free distilled water. The serum was mixed with 1× dilution buffer in a centrifuge tube to prepare the sample dilution solution. The sample dilution solution was incubated at 65°C for 30 minutes and then cooled to room temperature. Fifty microliters of the sample dilution solution was added to a 96-well plate, and 50 μL of assay buffer was added to each well. The solution was incubated at room temperature for 5 minutes. Fifty microliters of the reaction solution was added to each well, and the solution was incubated at room temperature for 20 minutes. The SEAP chemiluminescence value was read on a microplate reader (Tecan, Infinite 200).

[0546] The effect of the compound in inhibiting target gene expression is evaluated by measuring the SEAP expression level in serum. The test sample that can inhibit the SEAP expression level is selected as a nucleic acid drug.

[0547] On day 15 after modeling, each mouse was given a single subcutaneous administration of 200 μl of saline containing 3 mg / kg (mpk) RNAi agent, or 200 μl of saline without RNAi agent as a vehicle control, according to Tables 5 and 6. The results of the HDI model screening are shown in Tables 5 and 6.

[0548] Table 5 The efficacy of siRNA carrying the compounds of the present invention (DR005690, DR005921 and DR005922) is better than that of the positive compound DR001479

[0549] Table 6 The efficacy of siRNA (DR005691) carrying the compound of the present invention is better than that of the positive compound DR001479, and the efficacy of siRNA (DR005687) not carrying the compound of the present invention is equivalent to that of the positive compound DR001479

[0550] Example 8: Validation of the compounds of the present invention in a mouse HDI model

[0551] Referring to the procedure of Example 7, the compounds of the present invention were tested in another mouse HDI model. The only difference from Example 7 was that this experiment used siRNA targeting APOC3, while Example 7 used siRNA targeting ANGPTL3. The administration and results are shown in Table 7.

[0552] Table 7 The efficacy of siRNAs carrying compounds of the present invention (DR005682 and DR005684) is comparable to that of DR005681 carrying prior art IB

[0553] The above is a further detailed description of the present invention in conjunction with specific preferred embodiments, and the specific implementation of the present invention should not be considered to be limited to these descriptions. For those skilled in the art of the present invention, without departing from the concept of the present invention, several simple deductions or substitutions can be made, which should be considered to fall within the scope of protection of the present invention.

Claims

1. A compound of formula (X), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof: in, X1 is a chemical bond, O or S; X2 is a chemical bond or NR1; Y is a chemical bond or [C(R a )(R b )] 1-4 ; R1 is selected from C 1-20 Alkyl, C 1-20 Halogenated alkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl, 5-14 membered heteroaryl or -[C(O)] p -C 1-20 Alkylene-(OCH2CH2) n R x , which are optionally replaced by one or more radicals selected from halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl or 5-14 membered heteroaryl; wherein R1 is optionally deuterated, up to fully deuterated; R2 is selected from H, D or OL2; R is selected from H, D, -L-OR', -L-NR"R"', R b or -C(OL2)(R c )(R d ), which are optionally deuterated, up to fully deuterated; L1 and L2 are selected from H, a reactive phosphorus group or a protecting group; R a 、R b 、R c and R d independently selected from H, D, halogen, CN, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -LC 3-8 Cycloalkyl or -L-4-7 membered heterocyclic group; wherein R a 、R b 、R c and R d Optionally deuterated, up to fully deuterated; Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated; R', R" and R'" are independently selected from H, -C 1-6 Alkylene-OH, -C 1-6 Alkylene-NH2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2- 6 alkenyl, C 2-6 Alkynyl, -C 0-6 Alkylene-C 3-8 Cycloalkyl, -C 0-6 Alkylene-3-10 membered heterocyclic group, -C 0-6 Alkylene-C 6-10 Aryl or -C 0- 6-alkylene-5-14 membered heteroaryl, wherein the above groups are optionally substituted by one or more selected from halogen, CN, NO2, OH, NH2, C 1-6 Alkyl or C 1-6 or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group; R x Selected from H, D, OH, OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl; wherein R x Optionally deuterated, up to fully deuterated; n=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; p = 0 or 1; R s Selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated; m = 0, 1, 2, 3, or 4; Provided that one of R and R2 is an OL2 group.

2. The compound of formula (X) according to claim 1, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein X1 is a chemical bond or O, preferably O.

3. The compound of formula (X) according to claim 1 or 2, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein X2 is NR1; R1 is selected from C 1-20 Alkyl, C 1-20 Halogenated alkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl, preferably selected from C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl, more preferably selected from C 3-8 Cycloalkyl or 3-10 membered heterocyclic group, especially C 5-6 Cycloalkyl.

4. The compound of formula (X) according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein Y is a chemical bond.

5. The compound of formula (X) according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein Y is [C(R a )(R b )] 1-4 , preferably C(R a )(R b ).

6. The compound of formula (X) according to any one of claims 1 to 5, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R2 is selected from H or D.

7. The compound of formula (X) according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein One of R is -C(OL2)(R c )(R d ), which are optionally deuterated, up to fully deuterated.

8. The compound of formula (X) according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein One of L1 and L2 is a phosphoramidite (preferably -P(OCH2CH2CN)(N(iPr)2)), and the other is a di-p-methoxytrityl group (DMTr).

9. The compound of formula (X) according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R a 、R b 、R c and R d Independently selected from H, D, halogen, CN, -C 0-6 Alkyl-OR', -C 0-6 Alkyl-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, preferably H, D, halogen, CN, C 1-6 Alkyl or C 1-6 Haloalkyl; wherein R a 、R b 、R c and R d Optionally deuterated, up to fully deuterated.

10. A compound of formula (X) according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, selected from the following general formula: wherein each group is as defined in claims 1 to 9.

11. The compound of formula (X) according to claim 10, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, which is a compound of formula (I): in, X1 is a chemical bond, O or S; R1 is selected from C 1-20 Alkyl, C 1-20 Halogenated alkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl, 5-14 membered heteroaryl or -[C(O)] p -C 1-20 Alkylene-(OCH2CH2) n R x , which are optionally replaced by one or more radicals selected from halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl or 5-14 membered heteroaryl; wherein R1 is optionally deuterated, up to fully deuterated; Any of R is -C(OL2)(R c )(R d ), the other two are R b ; L1 and L2 are selected from H, a reactive phosphorus group or a protecting group; R a 、R b 、R c and R d independently selected from H, D, halogen, CN, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -LC 3-8 Cycloalkyl or -L-4-7 membered heterocyclic group; wherein R a 、R b 、R c and R d Optionally deuterated, up to fully deuterated; Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated; R', R" and R'" are independently selected from H, -C 1-6 Alkylene-OH, -C 1-6 Alkylene-NH2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2- 6 alkenyl, C 2-6 Alkynyl, -C 0-6 Alkylene-C 3-8 Cycloalkyl, -C 0-6 Alkylene-3-10 membered heterocyclic group, -C 0-6 Alkylene-C 6-10 Aryl or -C 0- 6-alkylene-5-14 membered heteroaryl, wherein the above groups are optionally substituted by one or more selected from halogen, CN, NO2, OH, NH2, C 1-6 Alkyl or C 1-6 or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group; R x Selected from H, D, OH, OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl; wherein R x Optionally deuterated, up to fully deuterated; n=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; p=0 or 1.

12. The compound of formula (I) according to claim 11, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein X1 is a chemical bond, O or S; R1 is cyclohexyl, which is optionally deuterated, up to fully deuterated; Any of R is -C(OL2)(R c )(R d ), the other two are R b ; L1 and L2 are selected from DMTr or -P(OCH2CH2CN)(N(iPr)2); R a 、R b 、R c and R d Independently selected from H or D.

13. The compound of formula (X) according to claim 10, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, which is a compound of formula (Ia), (Ib) or (Ic): in, X1 is a chemical bond, O or S; R1 is selected from C 1-20 Alkyl, C 1-20 Halogenated alkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl, 5-14 membered heteroaryl or -[C(O)] p -C 1-20 Alkylene-(OCH2CH2) n R x , which are optionally replaced by one or more radicals selected from halogen, CN, OR', SR', NR"R"', C(O)OR', C(S)OR', C(O)NR"R"', C(S)NR"R"', OC(O)R', OC(S)R', NR"C(O)R"', NR"C(S)R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 aryl or 5-14 membered heteroaryl; wherein R1 is optionally deuterated, up to fully deuterated; L1 and L2 are selected from H, a reactive phosphorus group or a protecting group; R a 、R b 、R c and R d independently selected from H, D, halogen, CN, -L-OR', -L-NR"R"', C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -LC 3-8 Cycloalkyl or -L-4-7 membered heterocyclic group; wherein R a 、R b 、R c and R d Optionally deuterated, up to fully deuterated; Where L is a chemical bond or C 1-6 Alkylene, which is optionally deuterated, up to fully deuterated; R', R" and R'" are independently selected from H, -C 1-6 Alkylene-OH, -C 1-6 Alkylene-NH2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2- 6 alkenyl, C 2-6 Alkynyl, -C 0-6 Alkylene-C 3-8 Cycloalkyl, -C 0-6 Alkylene-3-10 membered heterocyclic group, -C 0-6 Alkylene-C 6-10 Aryl or -C 0- 6-alkylene-5-14 membered heteroaryl, wherein the above groups are optionally substituted by one or more selected from halogen, CN, NO2, OH, NH2, C 1-6 Alkyl or C 1-6 or R", R'" and the nitrogen atom to which they are attached together form a 3-10 membered heterocyclic group; R x Selected from H, D, OH, OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl; wherein R x Optionally deuterated, up to fully deuterated; n=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; p = 0 or 1; Preferably, X1 is a chemical bond, O or S; R1 is selected from C 3-6 Cycloalkyl or 5-6 membered heterocyclyl, which is optionally deuterated up to fully deuterated; L1 and L2 are selected from DMTr or -P(OCH2CH2CN)(N(iPr)2); R a 、R b 、R c and R d Independently selected from H, D, halogen, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated; Preferably, X1 is a chemical bond, O or S; R1 is C 4-6 Cycloalkyl, which is optionally deuterated, up to fully deuterated; L1 and L2 are selected from DMTr or -P(OCH2CH2CN)(N(iPr)2); R a 、R b 、R c and R d Independently selected from H, D, halogen, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; Preferably, X1 is a chemical bond or O; R1 is cyclohexyl, which is optionally deuterated, up to fully deuterated; L1 and L2 are selected from DMTr or -P(OCH2CH2CN)(N(iPr)2); R a 、R b 、R c and R d Independently selected from H or D.

14. The compound of any one of claims 1 to 13, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein The compound is selected from the following:

15. An oligonucleotide comprising one or more compounds of formula (X') at the 5' and / or 3' end: in, R2 is selected from H, D or O R is selected from H, D, -L-OR', -L-NR"R"', R b or It is optionally deuterated, up to fully deuterated; The condition is that one of R and R2 exists group, in which represents the chemical bond connecting to the nucleotides of the oligonucleotide, and the terminal Connected to H, reactive phosphorus groups or protecting groups, Other groups are defined as defined in any one of claims 1 to 14.

16. The oligonucleotide of claim 15, wherein the compound of formula (X') is the following compound: wherein each group is defined as defined in any one of claims 1 to 15.

17. The oligonucleotide of claim 15, wherein the compound of formula (X') is the following compound: The order of 5'–>3' is from the to connect.

18. A double-stranded RNA having a sense strand and an antisense strand with fully complementary sequences, each strand having 14 to 30 nucleotides, the nucleotides being linked by phosphate groups, phosphorothioate groups, or other linking molecules, wherein the sense strand has the following structure: 5'(NT1) n1 -(NT2) n2 -(NT3) n3 3' in, NT1 is a compound of formula (X') according to any one of claims 15 to 17; NT2 is a modified or unmodified nucleotide; NT3 is a compound of formula (X') according to any one of claims 15 to 17; n1 is 0, 1, 2, or 3; n2 is an integer from 14 to 30; n3 is 0, 1, 2, or 3; And n1 and n3 are not 0 at the same time.

19. The double-stranded RNA according to claim 18, wherein n1 is 0, NT3 is selected from the compound of formula (I'), (Ia'), (Ib') or (Ic'), and n3 is 2.

20. The double-stranded RNA according to claim 18, wherein n1 is 2, NT1 is a compound of formula (I'), (Ia'), (Ib') or (Ic'), NT3 is selected from a compound of formula (I'), (Ia'), (Ib') or (Ic'), and n3 is 2.

21. The double-stranded RNA according to claim 18, wherein n1 is 1, NT1 is selected from the compound of formula (I'), (Ia'), (Ib') or (Ic'), NT3 is selected from the compound of formula (I'), (Ia'), (Ib') or (Ic'), and n3 is 1.

22. The double-stranded RNA according to claim 18, wherein n1 is 1, NT1 is selected from the compound of formula (I'), (Ia'), (Ib') or (Ic'), NT3 is selected from the compound of formula (I'), (Ia'), (Ib') or (Ic'), and n3 is 2.

23. The double-stranded RNA according to any one of claims 18 to 22, which is further conjugated to a ligand moiety comprising N-acetylgalactosamine, preferably conjugated to the ligand moiety at its sense strand, preferably conjugated to the 3' end of the sense strand, preferably conjugated to the ligand moiety at the 5' end of the sense strand.

24. The double-stranded RNA of claim 23, wherein the ligand comprises one or more GalNAc. The double-stranded RNA according to claim 24 , wherein a ligand comprising GalNAc is coupled to the 3′ end of the sense strand, and the ligand is selected from L96, GL6 or GL12, preferably GL6.

26. The double-stranded RNA according to any one of claims 18 to 25, which is selected from small interfering RNA (siRNA) and short hairpin RNA (shRNA).

27. A cell comprising the double-stranded RNA of any one of claims 18 to 26.

28. A pharmaceutical composition comprising the double-stranded RNA according to any one of claims 18 to 26, or the cell according to claim 27, and optionally a pharmaceutically acceptable carrier or excipient.

29. A kit comprising the double-stranded RNA according to any one of claims 18 to 26, or the cell according to claim 27.

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