Prodrugs for delivery of siRNA into cells

Abstract

The present invention provides an oligonucleotide comprising a compound of Formula (Ia), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof. The compound of formula (I) is contained inside the oligonucleotide, or at the 5 '-terminal and / or the 3'-terminal, for improving the ability of the double-stranded RNA to pass through the cell membrane. The invention also relates to a double-stranded RNA, a cell, a pharmaceutical composition and a kit comprising the oligonucleotide. # imgabs0 #

Description

Prodrugs for intracellular delivery of siRNA

[0001] The present invention claims priority to Chinese application No. 202211374509.3 filed on October 31, 2022, Chinese application No. 202310101722.5 filed on February 9, 2023, and Chinese application No. 202310552663.3 filed on May 16, 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 specifically relates to a masking group capable of enhancing the ability of double-stranded RNA to pass through cell membranes, such as a TXL-group in the structure of formula (I), and a compound of formula (I) in which the masking group is connected to a nucleotide, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof. Background Art

[0003] RNA interference is a phenomenon in which double-stranded RNA (dsRNA) induces the efficient and specific degradation of target mRNA.

[0004] Research has found that by preparing small interfering RNA (siRNA) in the form of a prodrug, the negative charge of the oligonucleotide phosphate group can be masked with a cell-cleavable masking group, thereby enhancing cell penetration. For example, WO2022147214A2 discloses a prodrug based on a disulfide-bonded ring.

[0005] However, prodrug approaches remain challenging, in part due to the difficulty in selecting the optimal masking group. For example, cellular cleavage of the masking group often produces products that are considered unfavorable or even toxic. Furthermore, the masking group must strike a balance between allowing absorption in the intestine and allowing cleavage in the blood or target cells.

[0006] Therefore, there is a need in the art to develop more prodrugs for the effective in vivo delivery of siRNA.

[0007] Summary of the Invention

[0008] In one aspect, the present invention provides an oligonucleotide comprising a compound of formula (II), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0009] Wherein, each variable is defined as follows.

[0010] In another aspect, the present invention provides an oligonucleotide comprising one, two or more compounds of formula (Ia), or pharmaceutically acceptable salts, tautomers or stereoisomers thereof:

[0011] Wherein, each variable is defined as follows.

[0012] In another aspect, the present invention provides a double-stranded RNA having a sense strand and an antisense strand, each strand having 14 to 30 nucleotides, the antisense strand comprising a sequence sufficiently complementary to the sense strand and the target mRNA, wherein the sense strand and / or antisense strand comprises one or more compounds of the aforementioned formula (II) or the aforementioned formula (Ia), or pharmaceutically acceptable salts, tautomers or stereoisomers thereof.

[0013] In another aspect, the present invention provides a cell containing the aforementioned double-stranded RNA.

[0014] In another aspect, the present invention provides a pharmaceutical composition comprising the aforementioned double-stranded RNA, the aforementioned cells, and optionally a pharmaceutically acceptable carrier or excipient.

[0015] In another aspect, the present invention provides a kit comprising the aforementioned double-stranded RNA, the aforementioned cell, or the aforementioned pharmaceutical composition.

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

[0017] Wherein, each variable is defined as follows.

[0018] Detailed Description of the Invention

[0019] definition

[0020] Chemical definition

[0021] Definitions of specific functional groups and chemical terms are described in more detail below.

[0022] 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.

[0023] “C 1-6 "Alkyl" refers to a straight or branched chain saturated hydrocarbon group having 1 to 6 carbon atoms. In some embodiments, 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).

[0024] “C 2-6 "Alkenyl" refers to a straight or branched chain hydrocarbon group having 2 to 6 carbon atoms and at least one carbon-carbon double bond. In some embodiments, 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 2-6 "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). An 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.

[0025] “C 2-6 "Alkynyl" refers to a straight or branched chain hydrocarbon group having 2 to 6 carbon atoms, at least one carbon-carbon triple bond, and optionally one or more carbon-carbon double bonds. In some embodiments, C 2-4 Alkynyl is preferred. 2-6Examples 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.

[0026] "Halo" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

[0027] Therefore, “C 1-6 "Haloalkyl" refers to the above-mentioned "C 1-6 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.

[0028] “C 3-10 "Cycloalkyl" refers to a non-aromatic cyclic hydrocarbon group having 3 to 10 ring carbon atoms and zero heteroatoms. In some embodiments, C 4-7 Cycloalkyl and C 3-6 Cycloalkyl is particularly preferred, more preferably 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.

[0029] "3-10 membered heterocyclyl" refers to a group 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 4-10 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 3-8 membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms; a 3-6 membered heterocyclyl is preferred, which is a 3-6 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 4-7 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 5-6 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, 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) include, but are not limited to, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothiophenyl, benzoxazolinonyl, 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. The heterocyclyl group may be optionally substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0030] “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.

[0031] "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. 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.

[0032] Alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and the like are defined herein as optionally substituted groups.

[0033] 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 、-NR bb 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;

[0034] 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;

[0035] 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;

[0036] 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;

[0037] R cc Each 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;

[0038] R ddEach 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)2Ree 、-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;

[0039] 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;

[0040] 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;

[0041] 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 1-6alkyl), -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 1-6 alkyl), 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 1-6alkyl)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.

[0042] 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 cc 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 dd group substituted, and wherein R aa 、R bb 、R cc and R dd As mentioned above.

[0043] Other definitions

[0044] 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.

[0045] The term "antisense strand" refers to a strand of an siRNA that includes a region that is completely, 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 completely, fully, or substantially complementary to a region that is the antisense strand as that term is defined herein.

[0046] The term "complementary region" refers to a region on the antisense strand that is completely, fully 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, in 5' and / or 3' ends within 5, 4, 3, 2 or 1 nucleotide. 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.

[0047] 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.

[0048] A polynucleotide that is "at least partially complementary," "sufficiently 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 the mRNA encoding PCSK9. The terms "complementary," "fully complementary," "sufficiently 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.

[0049] "Fully complementary" refers to the extent to which the sense strand only needs to be complementary to the antisense strand in order to maintain the overall double-stranded nature of the molecule. In other words, while perfect complementarity is generally desired, in some cases, particularly in the antisense strand, one or more, for example, 6, 5, 4, 3, 2, or 1 mismatches (relative to the target mRNA) may be included, but the sense and antisense strands can still maintain the overall double-stranded nature of the molecule.

[0050] "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 the Pol III promoter. Five to six Ts are then attached to serve as a transcriptional terminator for RNA polymerase III.

[0051] 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.

[0052] "Ribose" is a five-carbon aldose sugar composed of five carbon atoms. The carbon atom adjacent to oxygen is numbered 1' (abbreviated as 1' carbon atom or 1' end). The carbon atoms are numbered in a clockwise direction, with the base attached at 1' and the phosphate group attached at 5'. The structural formula and carbon atom numbering are as follows:

[0053] "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.

[0054] 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.

[0055] " 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.

[0056] A "protecting group," also known as a "blocking group," refers to any atom or group of atoms added to a molecule to prevent existing groups in the molecule from undergoing undesirable chemical reactions. 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, from 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.

[0057] 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).

[0058] 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:

[0059] 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), -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytrityl, preferably -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytrityl, more preferably -C(O)CH2CH2C(O)OH.

[0060] 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.

[0061] 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.

[0062] 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.

[0063] 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.

[0064] Compounds of the present invention

[0065] The present invention specifically relates to a compound of formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0066] in,

[0067] X1 is selected from OR a , OP1, SP1 or NR b R c ;

[0068] R a Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0069] R b and R c Independently selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration;

[0070] X2 is OH, OP1 or SP1, or X2 is a nucleoside moiety linked to P via the hydroxyl or sulfhydryl group at the 2', 3' or 5' end of the ribose;

[0071] X3 is independently selected from O or S;

[0072] T is selected from

[0073] Each R T1 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0074] Each R T2 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0075] Each R T3 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0076] Each R T4 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0077] m is 0, 1, 2, 3, 4 or 5;

[0078] n is 0, 1, 2, 3, 4 or 5;

[0079] p is 0, 1, 2, 3, 4, or 5;

[0080] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0081] R X1 Selected from H, halogen, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0082] L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by R#, R# is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0083] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0084] Ar is selected from C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl, the C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2, 3, 4 or 5 R*;

[0085] R* is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0086] wherein P1 is selected from 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), 4,4'-dimethoxytrityl, -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH, preferably -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH.

[0087] The present invention also relates to oligonucleotides comprising one, two or more compounds of formula (Ia), or pharmaceutically acceptable salts, tautomers or stereoisomers thereof; the present invention also relates to double-stranded RNA having a sense strand and an antisense strand, each strand having 14 to 30 nucleotides, the antisense strand comprising a sequence that is sufficiently complementary to the sense strand and the target mRNA, wherein the sense strand and / or antisense strand comprises one or more compounds of formula (Ia), or pharmaceutically acceptable salts, tautomers or stereoisomers thereof:

[0088] in,

[0089] Selected from OR a or NR b R c , or represents a chemical bond connecting to the hydroxyl or sulfhydryl group at the 5' end of the ribose of the adjacent nucleotide;

[0090] X1 is selected from OR a or NR b R c , or a chemical bond connected to the 2' or 3' end of the ribose of another adjacent nucleotide;

[0091] and OR when X1 is different from a or NRb R c ;

[0092] R a Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0093] R b and R c Independently selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration;

[0094] X3 is independently selected from O or S;

[0095] T is selected from

[0096] Each R T1 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0097] Each R T2 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0098] Each R T3 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0099] Each R T4 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6alkynyl, which is optionally deuterated, up to fully deuterated;

[0100] m is 0, 1, 2, 3, 4 or 5;

[0101] n is 0, 1, 2, 3, 4 or 5;

[0102] p is 0, 1, 2, 3, 4, or 5;

[0103] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0104] R X1 Selected from H, halogen, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0105] L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by R#, R# is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0106] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0107] Ar is selected from C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl, the C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2, 3, 4 or 5 R*;

[0108] R* is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, which is optionally deuterated, up to fully deuterated.

[0109] The present invention also relates to an oligonucleotide comprising a compound of formula (II), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0110] in,

[0111] X1 is selected from OR a or NR b R c ;

[0112] R a Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0113] R b and R c Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration;

[0114] X2 is the remainder of the oligonucleotide, which is linked to P through the hydroxyl or sulfhydryl group on the 2', 3' or 5' carbon atom of the ribose of the first nucleotide at the 5' end;

[0115] X3 is independently selected from O or S;

[0116] Each R T1 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0117] Each R T2 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0118] m is 0, 1, 2, 3, 4 or 5;

[0119] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0120] R X1 Selected from H, C 1-6Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0121] L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0122] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0123] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2, 3, 4 or 5 R*;

[0124] R*selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, which is optionally deuterated, up to fully deuterated.

[0125] The present invention also relates to oligonucleotides comprising one, two or more compounds of formula (Ia), or pharmaceutically acceptable salts, tautomers or stereoisomers thereof:

[0126] in,

[0127] Selected from OR a or NR b R c , or represents a chemical bond connecting to the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide;

[0128] X1 is a chemical bond connecting to the hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose of another adjacent nucleotide;

[0129] R a Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0130] R b and R c Independently selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and Rc Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration;

[0131] X3 is independently selected from O or S;

[0132] T is

[0133] Each R T1 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0134] Each R T2 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0135] m is 0, 1, 2, 3, 4 or 5;

[0136] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0137] R X1 Selected from H, halogen, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0138] L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by R#, R# is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0139] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0140] Ar is selected from C 3-10Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl, the C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2, 3, 4 or 5 R*;

[0141] R* is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, which is optionally deuterated, up to fully deuterated.

[0142] X1, X2, X3 and

[0143] In one embodiment, X1 is OR a In another embodiment, X1 is OP1; In another embodiment, X1 is SP1; In another embodiment, X1 is NR b R c In another embodiment, X1 is a hydroxyl or sulfhydryl-linked chemical bond connected to the 5' end of the ribose of another adjacent nucleotide; in another embodiment, X1 is a hydroxyl or sulfhydryl-linked chemical bond connected to the 2' end of the ribose of another adjacent nucleotide; in another embodiment, X1 is a hydroxyl or sulfhydryl-linked chemical bond connected to the 3' end of the ribose of another adjacent nucleotide.

[0144] In one embodiment, X1 is a chemical bond to a hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose sugar of another adjacent nucleotide.

[0145] In a more specific embodiment, X1 is OH; in another more specific embodiment, X1 is

[0146] In one embodiment, X2 is OH; in another embodiment, X2 is OP1; in another embodiment, X2 is SP1; in another embodiment, X2 is a nucleoside moiety that is linked to P via the hydroxyl or sulfhydryl group at the 2' end of the ribose; in another embodiment, X2 is a nucleoside moiety that is linked to P via the hydroxyl or sulfhydryl group at the 3' end of the ribose; in another embodiment, X2 is a nucleoside moiety that is linked to P via the hydroxyl or sulfhydryl group at the 5' end of the ribose.

[0147] In one embodiment, X2 is the remainder of the oligonucleotide, which is linked to P via a hydroxyl group or a sulfhydryl group on the 2', 3' or 5' carbon atom of the ribose sugar of the first nucleotide at the 5' end, for example, it is linked to P via a hydroxyl group or a sulfhydryl group on the 5' carbon atom of the ribose sugar of the first nucleotide at the 5' end.

[0148] In one embodiment, X3 is O; in another embodiment, X3 is S.

[0149] In one embodiment, OR a In another embodiment, NR b R c In another embodiment, is a chemical bond connected to the 5' end of the ribose of another adjacent nucleotide, which is a hydroxyl or sulfhydryl bond; in another embodiment, OR when X1 is different from a or NR b R c .

[0150] In one embodiment, It represents a chemical bond to the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide.

[0151] T

[0152] In one embodiment, T is In another embodiment, T is In another embodiment, T is In another embodiment, T is In another embodiment, T is

[0153] In one embodiment, T is In another embodiment, T is In another embodiment, T is

[0154] In a more specific embodiment, T is In another more specific embodiment, T is In another more specific embodiment, T is In another more specific embodiment, T is In another more specific embodiment, T is In another more specific embodiment, T is

[0155] RT1 、R T2 、R T3 、R T4 、R T , m, n and p

[0156] In one embodiment, R T1 is H; in another embodiment, R T1 is D; in another embodiment, R T1 is halogen; in another embodiment, R T1 is CN; in another embodiment, R T1 C 1-6 Alkyl groups, such as C 1-4 Alkyl; in another embodiment, R T1 C 1-6 Haloalkyl, such as C 1-4 haloalkyl; in another embodiment, R T1 C 1-6 haloalkyl; in another embodiment, R T1 C 2-6 alkenyl; in another embodiment, R T1 C 2-6 Alkynyl; in another embodiment, R T1 is a chain comprising GalNAc; in another embodiment, said R T1 Optionally deuterated, up to fully deuterated.

[0157] In one embodiment, R T2 is H; in another embodiment, R T2 is D; in another embodiment, R T2 is halogen; in another embodiment, R T2 is CN; in another embodiment, R T2 C 1-6 Alkyl groups, such as C 1-4 Alkyl; in another embodiment, R T2 C 1-6 Haloalkyl, such as C 1-4 haloalkyl; in another embodiment, R T2 C 2-6 alkenyl; in another embodiment, R T2 C 2-6 Alkynyl; In another embodiment, said R T2 Optionally deuterated, up to fully deuterated.

[0158] In one embodiment, R T3 is H; in another embodiment, R T3is D; in another embodiment, R T3 is halogen; in another embodiment, R T3 is CN; in another embodiment, R T3 C 1-6 Alkyl groups, such as C 1-4 Alkyl; in another embodiment, R T3 C 1-6 Haloalkyl, such as C 1-4 haloalkyl; in another embodiment, R T3 C 2-6 alkenyl; in another embodiment, R T3 C 2-6 Alkynyl; In another embodiment, said R T3 Optionally deuterated, up to fully deuterated.

[0159] In one embodiment, R T4 is H; in another embodiment, R T4 is D; in another embodiment, R T4 is halogen; in another embodiment, R T4 is CN; in another embodiment, R T4 C 1-6 Alkyl groups, such as C 1-4 Alkyl; in another embodiment, R T4 C 1-6 Haloalkyl, such as C 1-4 haloalkyl; in another embodiment, R T4 C 2-6 alkenyl; in another embodiment, R T4 C 2-6 Alkynyl; In another embodiment, said R T4 Optionally deuterated, up to fully deuterated.

[0160] In one embodiment, R T is H; in another embodiment, R T is D; in another embodiment, R T is CH3; in another embodiment, R T is a chain comprising GalNAc; in another embodiment, said R T Optionally deuterated, up to fully deuterated.

[0161] In one embodiment, m is 0; in another embodiment, m is 1; in another embodiment, m is 2; in another embodiment, m is 3; in another embodiment, m is 4; in another embodiment, m is 5.

[0162] In one embodiment, n is 0; in another embodiment, n is 1; in another embodiment, n is 2; in another embodiment, n is 3; in another embodiment, n is 4; in another embodiment, n is 5.

[0163] In one embodiment, p is 0; in another embodiment, p is 1; in another embodiment, p is 2; in another embodiment, p is 3; in another embodiment, p is 4; in another embodiment, p is 5.

[0164] X

[0165] In one embodiment, X is a chemical bond; in another embodiment, X is -O-; in another embodiment, X is -S-; in another embodiment, X is -C(O)-; in another embodiment, X is -C(O)O-; in another embodiment, X is -OC(O)-; in another embodiment, X is -OC(O)NR X1 -; In another embodiment, X is -NR X1 C(O)O-; in another embodiment, X is -NR X1 C(O)-; In another embodiment, X is -C(O)NR X1 -.

[0166] In a more specific embodiment, X is a chemical bond; in another more specific embodiment, X is -O-; in another more specific embodiment, X is -NHC(O)O-; in another more specific embodiment, X is -OC(O)NH-; in another more specific embodiment, X is -N(CH3)C(O)O-; in another more specific embodiment, X is -C(O)O-.

[0167] L

[0168] In one embodiment, L is -Ar-(CH2) 1-6 -O-, wherein each CH2 is optionally substituted by R#, Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom; in another embodiment, L is -Ar-(CH2) 1-4 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom.

[0169] In one embodiment, Ar is C 3-10 Cycloalkyl; in another embodiment, Ar is a 3-10 membered heterocyclyl; in another embodiment, Ar is C 6-10In another embodiment, Ar is a 5-14 membered heteroaryl, such as a 5-10 membered heteroaryl, such as a phenyl group; in another embodiment, the Ar may be optionally substituted with 1, 2, 3, 4 or 5 R*.

[0170] In a more specific embodiment, L is In another more specific embodiment, L is In another more specific embodiment, L is In another more specific embodiment, L is In another more specific embodiment, L is In another more specific embodiment, L is In another more specific embodiment, L is In another more specific embodiment, L is

[0171] R a 、R b and R c

[0172] In one embodiment, R a is H; in another embodiment, R a C 1-6 Alkyl groups, such as C 1-4 Alkyl; in another embodiment, R a C 1-6 Haloalkyl, such as C 1-4 haloalkyl; in another embodiment, R a C 2-6 alkenyl; in another embodiment, R a C 2-6 Alkynyl; In another embodiment, said R a Optionally deuterated, up to fully deuterated.

[0173] In one embodiment, R b is H; in another embodiment, R b C 1-6 Alkyl groups, such as C 1-4 Alkyl; in another embodiment, R b C 1-6 Haloalkyl, such as C 1-4 In another embodiment, the R b Can be optionally D, C 6-10 aryl or 5-10 membered heteroaryl, up to full deuteration.

[0174] In one embodiment, R cis H; in another embodiment, R c C 1-6 Alkyl groups, such as C 1-4 Alkyl; in another embodiment, R c C 1-6 Haloalkyl, such as C 1-4 In another embodiment, the R c Can be optionally D, C 6-10 aryl or 5-10 membered heteroaryl, up to full deuteration.

[0175] R X1

[0176] In one embodiment, R X1 is H; in another embodiment, R X1 C 1-6 Alkyl groups, such as C 1-4 Alkyl; in another embodiment, R X1 C 1-6 Haloalkyl, such as C 1-4 In another embodiment, the R X1 Optionally deuterated, up to fully deuterated.

[0177] R#

[0178] In one embodiment, R# is H; in another embodiment, R# is D; in another embodiment, R# is halogen; in another embodiment, R# is CN; in another embodiment, R# is C 1-6 Alkyl groups, such as C 1-4 Alkyl; in another embodiment, R# is C 1-6 Haloalkyl, such as C 1-4 In another embodiment, R# is C 2-6 In another embodiment, R# is C 2-6 Alkynyl; In another embodiment, said R# is optionally deuterated, up to fully deuterated.

[0179] R*

[0180] In one embodiment, R* is H; in another embodiment, R* is D; in another embodiment, R* is halogen; in another embodiment, R* is CN; in another embodiment, R* is C 1-6 Alkyl groups, such as C 1-4 Alkyl; in another embodiment, R* is C 1-6 Haloalkyl, such as C 1-4 In another embodiment, R* is C2-6 In another embodiment, R* is C 2-6 Alkynyl; in another embodiment, said R* is optionally deuterated, up to fully deuterated.

[0181] PG and P1

[0182] In one embodiment, PG is a protecting group; in another embodiment, PG is 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 (Cb), 2,2,2-trichloroethoxycarbonyl (Troc ... z), 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), 4,4'-dimethoxytrityl, -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH, preferably -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH.

[0183] In one embodiment, P1 is a protecting group; in another embodiment, P1 is 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 (Cb), 2,3-dichloroethoxycarbonyl (Troc), 2,4-dichloroethoxycarbonyl (Cb), 2,5-dichloroethoxycarbonyl (Cb), 2,6-dichloroethoxycarbonyl (Cb), 2,7-dichloroethoxycarbonyl (Cb), 2,8-dichloroethoxycarbonyl (Cb), 2,9-dichloroethoxycarbonyl (Cb), 2,10 ... z), 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), 4,4'-dimethoxytrityl, -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH, preferably -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH.

[0184] GalNAc-containing chains

[0185] In one embodiment, the GalNAc-containing chain is a conjugate group comprising formula (X'):

[0186] in,

[0187] Indicates the location of attachment to a biomolecule;

[0188] Q is independently H,

[0189] Wherein L1 is a chemical bond, -CH2-, -CH2CH2-, -C(O)-, -CH2O-, -CH2O-CH2CH2O- or -NHC(O)-(CH2NHC(O)) a -;

[0190] L2 is a chemical bond or -CH2CH2C(O)-;

[0191] L3 is a chemical bond, -(NHCH2CH2) b -、-(NHCH2CH2CH2) b - or -C(O)CH2-;

[0192] L4 is -(OCH2CH2) c -、-(OCH2CH2CH2) c -、-(OCH2CH2CH2CH2) c -、-(OCH2CH2CH2CH2CH2) c -or-NHC(O)-(CH2) d -;

[0193] Where a = 0, 1, 2 or 3;

[0194] b = 1, 2, 3, 4, or 5;

[0195] c = 1, 2, 3, 4, or 5;

[0196] d = 1, 2, 3, 4, 5, 6, 7, or 8;

[0197] A is a chemical bond, -CH2O- or -NHC(O)-;

[0198] A' is a chemical bond, -C(O)NH-, -NHC(O)- or -O(CH2CH2O) e -;

[0199] Where e is 1, 2, 3, 4 or 5;

[0200] B is a chemical bond, -CH2-, -C(O)-, -M-, -CH2-M- or -C(O)-M-;

[0201] Where M is

[0202] R1 and R2 together form -CH2CH2O- or -CH2CH(R)-O-, and R3 is H;

[0203] Or R1 and R3 together form -C 1-2 Alkylene-, and R2 is H;

[0204] wherein R is -OR', -CH2OR' or -CH2CH2OR', wherein R' is H, a hydroxyl protecting group or a solid phase support, and the hydroxyl protecting group is preferably -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytrityl;

[0205] m1 = 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0206] n1=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0207] In another aspect, the present invention relates to a ligand comprising N-acetylgalactosamine, wherein the conjugated group is represented by formula (I'):

[0208] in,

[0209] Indicates the location of attachment to a biomolecule;

[0210] Q is independently H,

[0211] Wherein L1 is a chemical bond, -CH2-, -CH2CH2-, -C(O)-, -CH2O-, -CH2O-CH2CH2O- or -NHC(O)-(CH2NHC(O)) a -;

[0212] L2 is a chemical bond or -CH2CH2C(O)-;

[0213] L3 is a chemical bond, -(NHCH2CH2) b -、-(NHCH2CH2CH2) b - or -C(O)CH2-;

[0214] L4 is -(OCH2CH2) c -、-(OCH2CH2CH2) c -、-(OCH2CH2CH2CH2) c -、-(OCH2CH2CH2CH2CH2) c -or-NHC(O)-(CH2) d -;

[0215] Where a = 0, 1, 2 or 3;

[0216] b = 1, 2, 3, 4, or 5;

[0217] c = 1, 2, 3, 4, or 5;

[0218] d = 1, 2, 3, 4, 5, 6, 7, or 8;

[0219] A is -CH2O- or -NHC(O)-;

[0220] A' is a chemical bond, -C(O)NH- or -NHC(O)-;

[0221] R1 and R2 together form -CH2CH2O- or -CH2CH(R)-O-, and R3 is H;

[0222] Or R1 and R3 together form -C1-2 Alkylene-, and R2 is H;

[0223] wherein R is -OR', -CH2OR' or -CH2CH2OR', wherein R' is H, a hydroxyl protecting group or a solid phase support, and the hydroxyl protecting group is preferably -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytrityl;

[0224] m1 = 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0225] n1=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0226] In another embodiment, the chain comprising GalNAc is a conjugate group comprising formula (X'), wherein

[0227] Q is independently H,

[0228] Wherein L1 is a chemical bond, -CH2-, -CH2CH2-, -C(O)-, -CH2O-, -CH2O-CH2CH2O- or -NHC(O)-(CH2NHC(O)) a -;

[0229] L2 is a chemical bond or -CH2CH2C(O)-;

[0230] L3 is a chemical bond, -(NHCH2CH2) b -、-(NHCH2CH2CH2) b - or -C(O)CH2-;

[0231] L4 is -(OCH2CH2) c -、-(OCH2CH2CH2) c -、-(OCH2CH2CH2CH2) c -、-(OCH2CH2CH2CH2CH2) c -or-NHC(O)-(CH2) d -;

[0232] Where a = 0, 1, 2 or 3;

[0233] b = 1, 2, 3, 4, or 5;

[0234] c = 1, 2, 3, 4, or 5;

[0235] d = 1, 2, 3, 4, 5, 6, 7, or 8;

[0236] A is a chemical bond, -CH2O- or -NHC(O)-;

[0237] A' is a chemical bond, -C(O)NH-, -NHC(O)- or -O(CH2CH2O) e -;

[0238] Where e is 1, 2, 3, 4 or 5;

[0239] B is a chemical bond, -CH2-, -M-, -CH2-M- or -C(O)-M-;

[0240] Where M is

[0241] R1 and R2 together form -CH2CH2O- or -CH2CH(R)-O-, and R3 is H;

[0242] Or R1 and R3 together form -C 1-2 Alkylene-, and R2 is H;

[0243] wherein R is -OR', -CH2OR' or -CH2CH2OR', wherein R' is H, a hydroxyl protecting group or a solid phase support, and the hydroxyl protecting group is preferably -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytrityl;

[0244] m1 = 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0245] n1=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0246] In another embodiment, the GalNAc-containing chain is a conjugate group comprising formula (X'), wherein:

[0247] Q is independently H,

[0248] Wherein L1 is a chemical bond, -CH2-, -CH2CH2-, -C(O)-, -CH2O-, -CH2O-CH2CH2O- or -NHC(O)-(CH2NHC(O)) a -;

[0249] L2 is a chemical bond or -CH2CH2C(O)-;

[0250] L3 is a chemical bond, -(NHCH2CH2) b -、-(NHCH2CH2CH2) b - or -C(O)CH2-;

[0251] L4 is -(OCH2CH2) c -、-(OCH2CH2CH2) c -、-(OCH2CH2CH2CH2) c -、-(OCH2CH2CH2CH2CH2) c -or-NHC(O)-(CH2) d -;

[0252] Where a = 0, 1, 2 or 3;

[0253] b = 1, 2, 3, 4, or 5;

[0254] c = 1, 2, 3, 4, or 5;

[0255] d = 1, 2, 3, 4, 5, 6, 7, or 8;

[0256] A is a chemical bond, -CH2O- or -NHC(O)-;

[0257] A' is -O(CH2CH2O) e -;

[0258] Where e is 1, 2, 3, 4 or 5;

[0259] B is a chemical bond, -CH2-, -C(O)-, -M-, -CH2-M- or -C(O)-M-;

[0260] Where M is

[0261] R1 and R2 together form -CH2CH2O- or -CH2CH(R)-O-, and R3 is H;

[0262] Or R1 and R3 together form -C 1-2 Alkylene-, and R2 is H;

[0263] wherein R is -OR', -CH2OR' or -CH2CH2OR', wherein R' is H, a hydroxyl protecting group or a solid phase support, and the hydroxyl protecting group is preferably -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytrityl;

[0264] m1 = 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0265] n1=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0266] Any technical solution or any combination thereof in any of the above specific embodiments may be combined with any technical solution or any combination thereof in any other specific embodiment. For example, any technical solution of X1 or any combination thereof may be combined with any technical solution of X2, X3, T, X, and L, or any combination thereof. The present invention is intended to include all combinations of these technical solutions, but due to space limitations, they are not listed one by one.

[0267] 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.

[0268] 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.

[0269] 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.

[0270] 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.

[0271] The present invention specifically relates to the following technical solutions:

[0272] A1. A compound of formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0273] in,

[0274] X1 is selected from OR a , OP1, SP1 or NR b R c ;

[0275] R a Selected from H, C 1-6 Alkyl, C1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0276] R b and R c Independently selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration;

[0277] X2 is OH, OP1 or SP1, or X2 is a nucleoside moiety linked to P via the hydroxyl or sulfhydryl group at the 2', 3' or 5' end of the ribose;

[0278] X3 is independently selected from O or S;

[0279] T is selected from

[0280] Each R T1 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0281] Each R T2 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0282] Each R T3 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0283] Each R T4 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0284] m is 0, 1, 2, 3, 4 or 5;

[0285] n is 0, 1, 2, 3, 4 or 5;

[0286] p is 0, 1, 2, 3, 4, or 5;

[0287] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0288] R X1 Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0289] L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by R#, R# is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0290] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0291] Ar is selected from C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl, the C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2, 3, 4 or 5 R*;

[0292] R* is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0293] wherein P1 is selected from a protecting group.

[0294] A2. The compound of formula (I) of technical solution A1, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein,

[0295] X1 is selected from OR a , OP1, SP1 or NR b R c ;

[0296] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0297] R b and R c Independently selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, and the R b and R c Can be optionally D, C 6-10 aryl substitution, which is optionally deuterated, up to fully deuterated;

[0298] X2 is OH, OP1 or SP1, or X2 is a nucleoside moiety linked to P via the hydroxyl or sulfhydryl group at the 2', 3' or 5' end of the ribose;

[0299] X3 is independently selected from O or S;

[0300] T is selected from

[0301] Each R T1 Independently selected from H, D, C 1-4 Alkyl, C 1-4 a haloalkyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0302] Each R T2 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0303] Each R T3 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0304] Each R T4 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0305] m is 0, 1, 2, or 3;

[0306] n is 0, 1, 2, or 3;

[0307] p is 0, 1, 2, or 3;

[0308] X is selected from a chemical bond, -O-, -S-, -OC(O)NR X1 -、-NR X1 C(O)O-, -C(O)O-, -OC(O)-, -NR X1 C(O)- or -C(O)NR X1 -;

[0309] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0310] L is -Ar-(CH2) 1-4 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0311] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0312] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0313] R*selected from C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0314] wherein P1 is selected from a protecting group.

[0315] A3. The compound of formula (I) of technical solution A1 or A2, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein,

[0316] X1 is selected from OR a , OP1, SP1 or NR b R c ;

[0317] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0318] R b and R c Independently selected from H, C 1-4 Alkyl, C 1-4haloalkyl, the R b and R c Optionally substituted with D, phenyl, up to full deuteration;

[0319] X2 is OH, OP1 or SP1, or X2 is a nucleoside moiety linked to P via the hydroxyl or sulfhydryl group at the 2', 3' or 5' end of the ribose;

[0320] X3 is independently selected from O or S;

[0321] T is selected from where R T is selected from H, D, CH3 or a chain comprising GalNAc, which is optionally deuterated, up to fully deuterated;

[0322] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0323] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0324] L is selected from

[0325] wherein P1 is selected from a protecting group.

[0326] A4. The compound of formula (I) according to any one of technical solutions A1-A3, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein:

[0327] X1 is selected from OH, OP1, SP1 or

[0328] X2 is OH, OP1 or SP1, or X2 is a nucleoside moiety linked to P via the hydroxyl or sulfhydryl group at the 2', 3' or 5' end of the ribose;

[0329] X3 is independently selected from O or S;

[0330] T is selected from

[0331] X is selected from a chemical bond, -O-, -NHC(O)O-, -OC(O)NH-, -N(CH3)C(O)O- or -C(O)O-;

[0332] L is selected from

[0333] wherein P1 is selected from a protecting group.

[0334] A5. The compound of formula (I) according to any one of technical solutions A1-A4, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the compound of formula (I) has the following structure:

[0335] Wherein, each group is as defined in Technical Solutions A1-A4.

[0336] A6. The compound of formula (I) of technical solution A5, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the compound of formula (I) has the structure of formula (II):

[0337] in,

[0338] X1 is selected from OR a or NR b R c ;

[0339] R a Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0340] R b and R c Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration;

[0341] X2 is OH, OP1 or SP1, or X2 is a nucleoside moiety linked to P via the hydroxyl or sulfhydryl group at the 2', 3' or 5' end of the ribose;

[0342] X3 is independently selected from O or S;

[0343] Each R T1 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0344] Each R T2Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0345] m is 0, 1, 2, 3, 4 or 5;

[0346] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0347] R X1 Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0348] L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0349] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0350] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2, 3, 4 or 5 R*;

[0351] R*selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0352] wherein P1 is selected from a protecting group.

[0353] A7. The compound of formula (I) of technical solution A6, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein,

[0354] X1 is selected from OR a or NR b R c ;

[0355] Ra Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0356] R b and R c Independently selected from H, C 1-4 Alkyl, C 1-4 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl substitution until complete deuteration;

[0357] X2 is OH, OP1 or SP1, or X2 is a nucleoside moiety linked to P via the hydroxyl or sulfhydryl group at the 2', 3' or 5' end of the ribose;

[0358] X3 is independently selected from O or S;

[0359] Each R T1 Independently selected from H, D, C 1-4 Alkyl, C 1-4 a haloalkyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0360] Each R T2 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0361] m is 0, 1, 2, or 3;

[0362] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0363] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0364] L is -Ar-(CH2) 1-4 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0365] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0366] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0367] R*selected from C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0368] wherein P1 is selected from a protecting group.

[0369] A8. The compound of formula (I) of technical solution A6 or A7, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein,

[0370] X1 is selected from OR a or NR b R c ;

[0371] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0372] R b and R c Independently selected from H, C 1-4 Alkyl, C 1-4 haloalkyl, the R b and R c Optionally substituted with D, phenyl, up to full deuteration;

[0373] X2 is OH, OP1 or SP1, or X2 is a nucleoside moiety linked to P via the hydroxyl or sulfhydryl group at the 2', 3' or 5' end of the ribose;

[0374] X3 is independently selected from O or S;

[0375] Selected from

[0376] where R T is selected from H, D, CH3 or a chain comprising GalNAc, which is optionally deuterated, up to fully deuterated;

[0377] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NRX1 C(O)- or -C(O)NR X1 -;

[0378] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0379] L is selected from

[0380] wherein P1 is selected from a protecting group.

[0381] A9. The compound of formula (I) according to any one of technical solutions A6-A8, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein:

[0382] X1 is selected from OH or

[0383] X2 is OH, OP1 or SP1, or X2 is a nucleoside moiety linked to P via the hydroxyl or sulfhydryl group at the 2', 3' or 5' end of the ribose;

[0384] X3 is independently selected from O or S;

[0385] Selected from

[0386] X is selected from a chemical bond, -O-, -NHC(O)O-, -OC(O)NH-, -N(CH3)C(O)O- or -C(O)O-;

[0387] L is selected from

[0388] wherein P1 is selected from a protecting group.

[0389] A10. The compound of formula (I) of technical solution A5, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the compound of formula (I) has a structure of formula (III) or (IV):

[0390] in,

[0391] X1 is selected from OR a , OP1, SP1 or NR b R c ;

[0392] R a Selected from H, C 1-6 Alkyl or C1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0393] R b and R c Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration;

[0394] X2 is OH, OP1 or SP1, or X2 is a nucleoside moiety linked to P via the hydroxyl or sulfhydryl group at the 2', 3' or 5' end of the ribose;

[0395] X3 is independently selected from O or S;

[0396] Each R T3 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0397] Each R T4 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0398] n is 0, 1, 2, 3, 4 or 5;

[0399] p is 0, 1, 2, 3, 4, or 5;

[0400] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0401] R X1 Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0402] L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0403] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0404] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0405] R*selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0406] wherein P1 is selected from a protecting group.

[0407] A11. The compound of formula (I) of technical solution A10, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein,

[0408] X1 is selected from OR a , OP1, SP1 or NR b R c ;

[0409] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0410] R b and R c Independently selected from H, C 1-4 Alkyl, C 1-4 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl substitution until complete deuteration;

[0411] X2 is OH, OP1 or SP1, or X2 is a nucleoside moiety linked to P via the hydroxyl or sulfhydryl group at the 2', 3' or 5' end of the ribose;

[0412] X3 is independently selected from O or S;

[0413] Each R T3 Independently selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0414] Each R T4 Independently selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0415] n is 0, 1, 2, or 3;

[0416] p is 0, 1, 2, or 3;

[0417] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0418] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0419] L is -Ar-(CH2) 1-4 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0420] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0421] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0422] R*selected from C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0423] wherein P1 is selected from a protecting group.

[0424] A12. The compound of formula (I) of technical solution A10 or A11, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the compound of formula (I) has a structure of formula (III-1) or (IV-1),

[0425] in,

[0426] X1 is selected from OR a , OP1, SP1 or NRb R c ;

[0427] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0428] R b and R c Independently selected from H, C 1-4 Alkyl, C 1-4 haloalkyl, the R b and R c Optionally substituted with D, phenyl, up to full deuteration;

[0429] X2 is OH, OP1 or SP1, or X2 is a nucleoside moiety linked to P via the hydroxyl or sulfhydryl group at the 2', 3' or 5' end of the ribose;

[0430] X3 is independently selected from O or S;

[0431] Each R T3 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0432] Each R T4 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0433] n is 0, 1, 2, or 3;

[0434] p is 0, 1, 2, or 3;

[0435] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0436] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0437] L is selected from

[0438] wherein P1 is selected from a protecting group.

[0439] A13. The compound of formula (I) of technical solution A12, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein,

[0440] X1 is selected from OH, OP1, SP1 or

[0441] X2 is OH, OP1 or SP1, or X2 is a nucleoside moiety linked to P via the hydroxyl or sulfhydryl group at the 2', 3' or 5' end of the ribose;

[0442] X3 is independently selected from O or S;

[0443] R T3 is selected from H, D or CH3, which are optionally deuterated, up to fully deuterated;

[0444] R T4 is H;

[0445] n is 3;

[0446] p is 0;

[0447] X is selected from a chemical bond, -O-, -NHC(O)O-, -OC(O)NH-, -N(CH3)C(O)O- or -C(O)O-;

[0448] L is selected from

[0449] wherein P1 is selected from a protecting group.

[0450] A14. The compound of formula (I) according to any one of technical solutions A1-A13, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the compound of formula (I) is selected from:

[0451] in,

[0452] X1 is selected from OH, OP1 or SP1 or Preferably OH or

[0453] X2 is OH, OP1 or SP1, or a chemical bond to the hydroxyl or sulfhydryl group of the 2' or 3' end of the ribose sugar of another nucleotide, nucleoside or oligonucleotide;

[0454] X3 is independently selected from O or S;

[0455] P1 is selected from a protecting group;

[0456] Preferably, the compound of formula (I) is selected from:

[0457] X2 is a nucleoside moiety that is linked to P via the hydroxyl group at the 2', 3' or 5' end of the ribose sugar.

[0458] A15. The compound of formula (I) according to any one of technical solutions A1 to A14, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein P1 is selected from 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, 1,2-di ... (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), 4,4'-dimethoxytrityl, -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH, preferably -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH.

[0459] A16. An oligonucleotide comprising one, two or more compounds of formula (Ia), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0460] in,

[0461] Indicates OR a or NR b R c , or a chemical bond connecting to the 5'-end hydroxyl or sulfhydryl group of the ribose of the adjacent nucleotide;

[0462] X1 is selected from OR a or NR b R c , or a chemical bond connected to the 2' or 3' end of the ribose of another adjacent nucleotide;

[0463] and OR when X1 is different from a or NR b R c ;

[0464] R a Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0465] R b and R c Independently selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration;

[0466] X3 is independently selected from O or S;

[0467] T is selected from

[0468] Each R T1 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0469] Each R T2 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0470] Each R T3 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0471] Each R T4 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0472] m is 0, 1, 2, 3, 4 or 5;

[0473] n is 0, 1, 2, 3, 4 or 5;

[0474] p is 0, 1, 2, 3, 4, or 5;

[0475] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0476] R X1 Selected from H, halogen, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0477] L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by R#, R# is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0478] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0479] Ar is selected from C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl, the C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2, 3, 4 or 5 R*;

[0480] R* is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, which is optionally deuterated, up to fully deuterated.

[0481] A17. The oligonucleotide of technical solution A16, wherein

[0482] It represents the chemical bond connecting to the hydroxyl or sulfhydryl group at the 5' end of the ribose sugar of the adjacent nucleotide;

[0483] X1 is selected from OR a or NR b R c , or a chemical bond connected to the 2' or 3' end of the ribose of another adjacent nucleotide;

[0484] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0485] R b and R c Independently selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, and the R b and R c Can be optionally D, C 6-10 aryl substitution, which is optionally deuterated, up to fully deuterated;

[0486] X2 and X3 are independently selected from O or S;

[0487] T is selected from

[0488] Each R T1 Independently selected from H, D, C 1-4 Alkyl, C 1-4 a haloalkyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0489] Each R T2 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0490] Each R T3 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0491] Each R T4 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0492] m is 0, 1, 2, or 3;

[0493] n is 0, 1, 2, or 3;

[0494] p is 0, 1, 2, or 3;

[0495] X is selected from a chemical bond, -O-, -S-, -OC(O)NR X1 -、-NR X1 C(O)O-, -C(O)O-, -OC(O)-, -NR X1 C(O)- or -C(O)NR X1 -;

[0496] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0497] L is -Ar-(CH2) 1-4 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0498] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0499] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0500] R*selected from C 1-4 Alkyl or C 1-4 Haloalkyl, which is optionally deuterated, up to fully deuterated.

[0501] A18. The oligonucleotide of technical solution A16 or A17, wherein

[0502] It represents the chemical bond connecting to the hydroxyl or sulfhydryl group at the 5' end of the ribose sugar of the adjacent nucleotide;

[0503] X1 is selected from OR a or NR b R c , or a chemical bond connected to the 2' or 3' end of the ribose of another adjacent nucleotide;

[0504] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0505] R band R c Independently selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, the R b and R c Optionally substituted with D, phenyl, up to full deuteration;

[0506] X3 is independently selected from O or S;

[0507] T is selected from where R T is selected from H, D, CH3 or a chain comprising GalNAc, which is optionally deuterated, up to fully deuterated;

[0508] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0509] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0510] L is selected from

[0511] A19. The oligonucleotide of any one of technical solutions A16-A18, wherein

[0512] It represents the chemical bond connecting to the hydroxyl or sulfhydryl group at the 5' end of the ribose sugar of the adjacent nucleotide;

[0513] X1 is selected from OH or or a chemical bond connected to the hydroxyl or sulfhydryl group of the 2' or 3' end of the ribose sugar of another adjacent nucleotide;

[0514] X3 is selected from O or S;

[0515] T is selected from

[0516] X is selected from a chemical bond, -O-, -NHC(O)O-, -OC(O)NH-, -N(CH3)C(O)O- or -C(O)O-;

[0517] L is selected from

[0518] A20. The oligonucleotide of any one of technical solutions A16-A19, wherein the compound of formula (Ia) is selected from the following structures:

[0519] Wherein, each group is as defined in technical solutions 16-19.

[0520] A21. The oligonucleotide of technical solution A20, wherein the compound of formula (Ia) has the structure of formula (IIa):

[0521] in,

[0522] It represents the chemical bond connecting to the hydroxyl or sulfhydryl group at the 5' end of the ribose sugar of the adjacent nucleotide;

[0523] X1 is selected from OR a or NR b R c ;

[0524] R a Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0525] R b and R c Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration;

[0526] X3 is independently selected from O or S;

[0527] Each R T1 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0528] Each R T2 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0529] m is 0, 1, 2, or 3;

[0530] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0531] R X1 Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0532] L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0533] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0534] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0535] R*selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, which is optionally deuterated, up to fully deuterated.

[0536] A22. The oligonucleotide of technical solution A21, wherein

[0537] It represents the chemical bond connecting to the hydroxyl or sulfhydryl group at the 5' end of the ribose sugar of the adjacent nucleotide;

[0538] X1 is selected from OR a or NR b R c ;

[0539] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0540] R b and R c Independently selected from H, C 1-4 Alkyl, C1-4 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl substitution until complete deuteration;

[0541] X3 is independently selected from O or S;

[0542] Each R T1 Independently selected from H, D, C 1-4 Alkyl, C 1-4 a haloalkyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0543] Each R T2 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0544] m is 0, 1, or 2;

[0545] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0546] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0547] L is -Ar-(CH2) 1-4 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0548] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0549] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0550] R*selected from C 1-4 Alkyl or C 1-4 Haloalkyl, which is optionally deuterated, up to fully deuterated.

[0551] A23. The oligonucleotide of technical solution A21 or A22, wherein

[0552] It represents the chemical bond connecting to the hydroxyl or sulfhydryl group at the 5' end of the ribose sugar of the adjacent nucleotide;

[0553] X1 is selected from OR a or NR b R c ;

[0554] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0555] R b and R c Independently selected from H, C 1-4 Alkyl, C 1-4 haloalkyl, the R b and R c optionally substituted with D, phenyl, which is optionally deuterated, up to fully deuterated;

[0556] X3 is independently selected from O or S;

[0557] Selected from

[0558] where R T is selected from H, D, CH3 or a chain comprising GalNAc, which is optionally deuterated, up to fully deuterated;

[0559] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0560] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0561] L is selected from

[0562] A24. The oligonucleotide of any one of technical solutions A21-A23, wherein

[0563] It represents the chemical bond connecting to the hydroxyl or sulfhydryl group at the 5' end of the ribose sugar of the adjacent nucleotide;

[0564] X1 is selected from OH or

[0565] X3 is selected from O or S;

[0566] Selected from

[0567] X is selected from a chemical bond, -O-, -NHC(O)O-, -OC(O)NH-, -N(CH3)C(O)O- or -C(O)O-;

[0568] L is selected from

[0569] A25. The oligonucleotide of any one of technical solutions A16-A24, wherein the compound of formula (Ia) is selected from the following structures:

[0570] in,

[0571] represents a chemical bond connected to the hydroxyl or sulfhydryl group at the 5' end of the ribose of the adjacent nucleotide; X1 is selected from OH or

[0572] X3 is selected from O or S;

[0573] Preferably, the compound of formula (Ia) is selected from:

[0574] It represents the chemical bond connecting to the 5'-end hydroxyl group of the ribose sugar of the adjacent nucleotide.

[0575] A26. The oligonucleotide of technical solution A20, wherein the compound of formula (Ia) has a structure of formula (IIIa) or (IVa):

[0576] in,

[0577] Selected from OR a or NR b R c , or represents a chemical bond connecting to the hydroxyl or sulfhydryl group at the 5' end of the ribose of the adjacent nucleotide;

[0578] X1 is selected from OR a or NR b R c, or a chemical bond connected to the 2' or 3' end of the ribose of another adjacent nucleotide;

[0579] and OR when X1 is different from a or NR b R c ;

[0580] R a Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0581] R b and R c Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration;

[0582] X3 is independently selected from O or S;

[0583] Each R T3 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0584] Each R T4 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0585] n is 0, 1, 2, 3, 4 or 5;

[0586] p is 0, 1, 2, 3, 4, or 5;

[0587] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0588] R X1 Selected from H, C 1-6 Alkyl or C1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0589] L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0590] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0591] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0592] R*selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, which is optionally deuterated, up to fully deuterated.

[0593] A27. The oligonucleotide of technical solution A26, wherein

[0594] Selected from OR a or NR b R c , or represents a chemical bond connecting to the hydroxyl or sulfhydryl group at the 5' end of the ribose of the adjacent nucleotide;

[0595] X1 is selected from OR a or NR b R c , or a chemical bond connected to the 2' or 3' end of the ribose of another adjacent nucleotide;

[0596] and OR when X1 is different from a or NR b R c ;

[0597] R a Selected from H, C 1-4 Alkyl or C 1-4 alkyl halide;

[0598] R b and R c Independently selected from H, C 1-4 Alkyl, C 1-4 haloalkyl, the R b and Rc Optionally C 6-10 aryl substitution;

[0599] X2 and X3 are independently selected from O or S;

[0600] Each R T3 Independently selected from H, C 1-6 Alkyl or C 1-6 alkyl halide;

[0601] Each R T4 Independently selected from H, C 1-6 Alkyl or C 1-6 alkyl halide;

[0602] n is 0, 1, 2, or 3;

[0603] p is 0, 1, 2, or 3;

[0604] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0605] R X1 Selected from H, C 1-4 Alkyl or C 1-4 alkyl halide;

[0606] L is -Ar-(CH2) 1-4 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# being selected from H, C 1-4 Alkyl or C 1-4 alkyl halide;

[0607] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0608] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0609] R*selected from C 1-4 Alkyl or C 1-4 Halogenated alkyl.

[0610] A28. The oligonucleotide of technical solution A26 or A27, wherein the compound of formula (Ia) has structure (III-1a) or (IV-1a):

[0611] in,

[0612] Selected from OR a or NR b R c , or represents a chemical bond connecting to the hydroxyl or sulfhydryl group at the 5' end of the ribose of the adjacent nucleotide;

[0613] X1 is selected from OR a or NR b R c , or a chemical bond connected to the 2' or 3' end of the ribose of another adjacent nucleotide;

[0614] and OR when X1 is different from a or NR b R c ;

[0615] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0616] R b and R c Independently selected from H, C 1-4 Alkyl, C 1-4 haloalkyl, the R b and R c Optionally substituted with D, phenyl, up to full deuteration;

[0617] X3 is independently selected from O or S;

[0618] Each R T3 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0619] Each R T4 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0620] n is 0, 1, 2, or 3;

[0621] p is 0, 1, 2, or 3;

[0622] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NRX1 C(O)- or -C(O)NR X1 -;

[0623] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0624] L is selected from

[0625] A29. The oligonucleotide of technical solution A28, wherein

[0626] Selected from OH or or represents a chemical bond to the hydroxyl or sulfhydryl group at the 5' end of the ribose sugar of the adjacent nucleotide;

[0627] X1 is selected from OH or or a chemical bond connected to the hydroxyl or sulfhydryl group of the 2' or 3' end of the ribose sugar of another adjacent nucleotide;

[0628] and and X1 are not OH or

[0629] X3 is selected from O or S;

[0630] R T3 is selected from H, D or CH3, which are optionally deuterated, up to fully deuterated;

[0631] R T4 is H;

[0632] n is 3;

[0633] p is 0;

[0634] X is selected from a chemical bond, -O-, -NHC(O)O-, -OC(O)NH-, -N(CH3)C(O)O- or -C(O)O-;

[0635] L is selected from

[0636] A30. The oligonucleotide of any one of technical solutions A16-A20 and A26-A29, wherein the compound of formula (Ia) is selected from the following structures:

[0637] in,

[0638] Selected from OH or or represents a chemical bond to the hydroxyl or sulfhydryl group at the 5' end of the ribose sugar of the adjacent nucleotide;

[0639] X1 is selected from OH or or a chemical bond connected to the hydroxyl or sulfhydryl group of the 2' or 3' end of the ribose sugar of another adjacent nucleotide;

[0640] and and X1 are not OH or

[0641] X3 is selected from O or S.

[0642] A31. The oligonucleotide of any one of technical solutions A16-A30, which has 14 to 30 nucleotides.

[0643] A32. The oligonucleotide of any one of technical solutions A16-A30, which comprises a compound of formula (Ia), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, at the 5′ end.

[0644] A33. The oligonucleotide of any one of technical solutions A16-A30, which comprises a compound of formula (Ia), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof at the 3' end.

[0645] A34. The oligonucleotide of any one of technical solutions A16-A30, which comprises a compound of formula (Ia) at the 5' end and the 3' end, respectively, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[0646] A35. The oligonucleotide of any one of technical solutions A16-A20, A26-A30, which comprises one or more compounds of formula (I'), or pharmaceutically acceptable salts, tautomers or stereoisomers thereof, inside the oligonucleotide.

[0647] A36. A double-stranded RNA having a sense strand and an antisense strand, each strand having 14 to 30 nucleotides, the antisense strand comprising a sequence that is sufficiently complementary to the sense strand and the target mRNA, wherein the sense strand and / or antisense strand comprises one or more compounds of formula (Ia) described in any one of technical solutions A16-A35, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[0648] A37. The double-stranded RNA of technical solution A36, wherein the sense strand comprises a compound of formula (Ia) according to any one of technical solutions A16 to A35 at the 5′ end, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[0649] A38. The double-stranded RNA of technical solution A36 or A37, wherein the sense strand comprises a compound of formula (Ia) according to any one of technical solutions A16 to A35 at the 3' end, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[0650] A39. The double-stranded RNA of any one of technical solutions A36-A38, wherein the sense strand comprises a compound of formula (Ia) according to any one of technical solutions A16-A35 at the 5' end and the 3' end, respectively, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[0651] A40. The double-stranded RNA of any one of technical solutions A36-A39, wherein the sense strand comprises one or more compounds of formula (I') according to any one of technical solutions A16-A20, A26-A30, or pharmaceutically acceptable salts, tautomers or stereoisomers thereof inside the oligonucleotide.

[0652] A41. The double-stranded RNA of any one of technical solutions A36-A40, wherein the antisense strand comprises a compound of formula (Ia) according to any one of technical solutions A16-A35 at the 5' end, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[0653] A42. The double-stranded RNA of any one of technical solutions A36-A41, wherein the antisense strand comprises a compound of formula (Ia) according to any one of technical solutions A16-A35 at the 3' end, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[0654] A43. The double-stranded RNA of any one of technical solutions A36-A42, wherein the antisense strand comprises a compound of formula (Ia) according to any one of technical solutions A16-A35 at the 5' end and the 3' end, respectively, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[0655] A44. The double-stranded RNA of any one of technical solutions A36-A43, wherein the antisense strand comprises one or more compounds of formula (I') according to any one of technical solutions A16-A20, A26-A30, or pharmaceutically acceptable salts, tautomers or stereoisomers thereof inside the oligonucleotide.

[0656] A45. The double-stranded RNA of any one of technical solutions A36-A44, which is selected from small interfering RNA (siRNA) and short hairpin RNA (shRNA).

[0657] A46. A vector comprising a nucleotide sequence encoding the double-stranded RNA described in any one of the aforementioned technical solutions A36-A45.

[0658] A47. A cell containing the double-stranded RNA as described in any one of technical solutions A36-A45 or the vector as described in technical solution 54.

[0659] A48. A pharmaceutical composition comprising the double-stranded RNA as described in any one of technical solutions A36-A45, the vector as described in technical solution 46, or the cell as described in technical solution 47, and optionally a pharmaceutically acceptable carrier or excipient.

[0660] A49. A kit comprising the double-stranded RNA as described in any one of technical solutions A36-A45, the vector as described in claim 46, or the cell as described in technical solution 47.

[0661] A50. A method for improving the effectiveness of siRNA in a cell, comprising the step of introducing the double-stranded RNA described in any one of technical solutions A36-A45 or the vector described in technical solution A46 into the cell.

[0662] A51. A compound of formula (IIIb) or (IVb), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0663] in,

[0664] Each R T3 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0665] Each R T4 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0666] n is 0, 1, 2, 3, 4 or 5;

[0667] p is 0, 1, 2, 3, 4, or 5;

[0668] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0669] R X1 Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0670] L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0671] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0672] R*selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0673] PG is selected from protecting groups.

[0674] A52. The compound of technical solution A51, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein

[0675] Each R T3 Independently selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0676] Each R T4 Independently selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0677] n is 0, 1, 2, or 3;

[0678] p is 0, 1, 2, or 3;

[0679] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0680] R X1 Selected from H, C1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0681] L is -Ar-(CH2) 1-4 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0682] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0683] R*selected from C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0684] PG is selected from protecting groups.

[0685] A53. The compound of technical solution A51 or A52, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein PG is selected from 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), di-p-methoxytrityl (DMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzyloxymethyl (PMBM), 4,4'-dimethoxytrityl, -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH, preferably -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH.

[0686] A54. The compound of any one of technical solutions A51-A53, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the compound of formula (IIIb) or (IVb) has the structure of formula (III-1b) or (IV-1b),

[0687] in,

[0688] Each R T3 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0689] Each R T4 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0690] n is 0, 1, 2, or 3;

[0691] p is 0, 1, 2, or 3;

[0692] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0693] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0694] L is selected from

[0695] PG is a protecting group selected from DMTr, -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH.

[0696] A55. The compound of technical solution A54, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein

[0697] R T3 is selected from H, D or CH3, which are optionally deuterated, up to fully deuterated;

[0698] R T4 Selected from H or D;

[0699] n is 3;

[0700] p is 0;

[0701] X is selected from a chemical bond, -O-, -NHC(O)O-, -OC(O)NH-, -N(CH3)C(O)O- or -C(O)O-;

[0702] L is selected from

[0703] PG is selected from DMTr, -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH.

[0704] A56. The compound of any one of technical solutions A51-A55, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the compound of formula (IIIb) or (IVb) is selected from:

[0705] in,

[0706] PG is selected from protecting groups, preferably hydroxy protecting groups, 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 (

[0015] The following are examples of the present invention:

[0016] wherein the present invention comprises an amino group comprising: a 1,2-diphenylmethyl group, a 2,2,2-trichloroethoxymethyl group, a 2-methoxyethoxymethyl group, a methylthiomethyl group, a p-methoxybenzyl group, a 4,4'-dimethoxytrityl group, a -P(OCH2CH2CN)(N(iPr)2) or a -C(O)CH2CH2C(O)OH group, preferably a -P(OCH2CH2CN)(N(iPr)2) or a -C(O)CH2CH2C(O)OH group.

[0707] A57. The compound of formula (I) according to any one of technical solutions A1-A3 and A6-A8 or the oligonucleotide according to any one of technical solutions A16-18 and A21-23, wherein the chain containing GalNAc is a conjugated group represented by formula (X'):

[0708] in,

[0709] Indicates the location of attachment to a biomolecule;

[0710] Q is independently H,

[0711] Wherein L1 is a chemical bond, -CH2-, -CH2CH2-, -C(O)-, -CH2O-, -CH2O-CH2CH2O- or -NHC(O)-(CH2NHC(O)) a -;

[0712] L2 is a chemical bond or -CH2CH2C(O)-;

[0713] L3 is a chemical bond, -(NHCH2CH2) b -、-(NHCH2CH2CH2) b - or -C(O)CH2-;

[0714] L4 is -(OCH2CH2) c -、-(OCH2CH2CH2) c -、-(OCH2CH2CH2CH2) c -、-(OCH2CH2CH2CH2CH2) c -or-NHC(O)-(CH2) d -;

[0715] Where a = 0, 1, 2 or 3;

[0716] b = 1, 2, 3, 4, or 5;

[0717] c = 1, 2, 3, 4, or 5;

[0718] d = 1, 2, 3, 4, 5, 6, 7, or 8;

[0719] A is a chemical bond, -CH2O- or -NHC(O)-;

[0720] A' is a chemical bond, -C(O)NH-, -NHC(O)- or -O(CH2CH2O) e -;

[0721] Where e is 1, 2, 3, 4 or 5;

[0722] B is a chemical bond, -CH2-, -C(O)-, -M-, -CH2-M- or -C(O)-M-;

[0723] Where M is

[0724] R1 and R2 together form -CH2CH2O- or -CH2CH(R)-O-, and R3 is H;

[0725] Or R1 and R3 together form -C 1-2 Alkylene-, and R2 is H;

[0726] wherein R is -OR', -CH2OR' or -CH2CH2OR', wherein R' is H, a hydroxyl protecting group or a solid phase support, and the hydroxyl protecting group is preferably -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytrityl;

[0727] m1 = 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0728] n1=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0729] The present invention also specifically relates to the following technical solutions:

[0730] B1. An oligonucleotide comprising a compound of formula (II), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0731] in,

[0732] X1 is selected from OR a or NR b R c ;

[0733] R a Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0734] R b and R c Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration;

[0735] X2 is the remainder of the oligonucleotide, which is linked to P through the hydroxyl or sulfhydryl group on the 2', 3' or 5' carbon atom of the ribose of the first nucleotide at the 5' end;

[0736] X3 is independently selected from O or S;

[0737] Each R T1 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0738] Each R T2 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0739] m is 0, 1, 2, 3, 4 or 5;

[0740] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0741] R X1 Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0742] L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0743] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0744] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2, 3, 4 or 5 R*;

[0745] R*selected from C 1-6 Alkyl, C1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, which is optionally deuterated, up to fully deuterated.

[0746] B2. The oligonucleotide of technical solution B1, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein

[0747] X1 is selected from OR a or NR b R c ;

[0748] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0749] R b and R c Independently selected from H, C 1-4 Alkyl, C 1-4 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl substitution until complete deuteration;

[0750] X2 is the remainder of the oligonucleotide, which is linked to P through the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose of the first nucleotide at the 5' end;

[0751] X3 is independently selected from O or S;

[0752] Each R T1 Independently selected from H, D, C 1-4 Alkyl, C 1-4 a haloalkyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0753] Each R T2 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0754] m is 0, 1, 2, or 3;

[0755] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0756] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0757] L is -Ar-(CH2)-O-, wherein CH2 may be optionally substituted by 1 or 2 R#, R# being selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0758] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0759] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0760] R*selected from C 1-4 Alkyl or C 1-4 Haloalkyl, which is optionally deuterated, up to fully deuterated.

[0761] B3. The oligonucleotide of technical solution B1 or B2, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein

[0762] X1 is selected from OR a or NR b R c ;

[0763] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0764] R b and R c Independently selected from H, C 1-4 Alkyl, C 1-4 haloalkyl, the R b and R c Optionally substituted with D, phenyl, up to full deuteration;

[0765] X2 is the remainder of the oligonucleotide, which is linked to P through the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose of the first nucleotide at the 5' end;

[0766] X3 is independently selected from O or S;

[0767] Selected from

[0768] where R T is selected from H, D, CH3 or a chain comprising GalNAc, which is optionally deuterated, up to fully deuterated;

[0769] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0770] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0771] L is selected from

[0772] B4. The oligonucleotide of any one of technical solutions B1-B3, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein

[0773] X1 is selected from OH or

[0774] X2 is the remainder of the oligonucleotide, which is linked to P through the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose of the first nucleotide at the 5' end;

[0775] X3 is independently selected from O or S;

[0776] Selected from

[0777] X is selected from a chemical bond, -O-, -NHC(O)O-, -OC(O)NH-, -N(CH3)C(O)O- or -C(O)O-;

[0778] L is selected from

[0779] B5. The oligonucleotide of any one of technical solutions B1-B4, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the compound of formula (II) is selected from:

[0780] in,

[0781] X1 is selected from OH or

[0782] X2 is the remainder of the oligonucleotide, which is linked to P via the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the first nucleotide at the 5' end;

[0783] X3 is independently selected from O or S;

[0784] Preferably, the compound of formula (I) is selected from:

[0785] X2 is the remainder of the oligonucleotide, which is linked to P via the hydroxyl group on the 5' carbon atom of the ribose sugar of the first nucleotide at the 5' end.

[0786] B6. An oligonucleotide comprising one, two or more compounds of formula (Ia), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0787] in,

[0788] Selected from OR a or NR b R c , or represents a chemical bond connecting to the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide;

[0789] X1 is a chemical bond connecting to the hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose of another adjacent nucleotide;

[0790] R a Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0791] R b and R c Independently selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration;

[0792] X3 is independently selected from O or S;

[0793] T is

[0794] Each R T1 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6Halogenated alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0795] Each R T2 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0796] m is 0, 1, 2, 3, 4 or 5;

[0797] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0798] R X1 Selected from H, halogen, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0799] L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by R#, R# is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0800] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0801] Ar is selected from C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl, the C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2, 3, 4 or 5 R*;

[0802] R* is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6Alkynyl, which is optionally deuterated, up to fully deuterated.

[0803] B7. The oligonucleotide of technical solution B6, wherein

[0804] Selected from OR a or NR b R c , or represents a chemical bond connecting to the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide;

[0805] X1 is a chemical bond connecting to the hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose of another adjacent nucleotide;

[0806] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0807] R b and R c Independently selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, and the R b and R c Can be optionally D, C 6-10 aryl substitution, which is optionally deuterated, up to fully deuterated;

[0808] X2 and X3 are independently selected from O or S;

[0809] T is selected from

[0810] Each R T1 Independently selected from H, D, C 1-4 Alkyl, C 1-4 a haloalkyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0811] Each R T2 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0812] m is 0, 1, 2, or 3;

[0813] X is selected from a chemical bond, -O-, -S-, -OC(O)NR X1 -、-NR X1 C(O)O-, -C(O)O-, -OC(O)-, -NR X1 C(O)- or -C(O)NR X1 -;

[0814] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0815] L is -Ar-(CH2)-O-, wherein CH2 may be optionally substituted by 1 or 2 R#, R# being selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0816] Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;

[0817] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0818] R*selected from C 1-4 Alkyl or C 1-4 Haloalkyl, which is optionally deuterated, up to fully deuterated.

[0819] B8. The oligonucleotide of technical solution B6 or B7, wherein

[0820] Selected from OR a or NR b R c , or represents a chemical bond connecting to the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide;

[0821] X1 is a chemical bond connecting to the hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose of another adjacent nucleotide;

[0822] R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0823] R b and R c Independently selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, the R b and R c Optionally substituted with D, phenyl, up to full deuteration;

[0824] X3 is independently selected from O or S;

[0825] T is selected from where RT is selected from H, D, CH3 or a chain comprising GalNAc, which is optionally deuterated, up to fully deuterated;

[0826] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0827] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0828] L is selected from

[0829] B9. The oligonucleotide of any one of technical solutions B6-B8, wherein

[0830] It represents a chemical bond to the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide;

[0831] X1 is a chemical bond connecting to the hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose of another adjacent nucleotide;

[0832] X3 is selected from O or S;

[0833] T is selected from

[0834] X is selected from a chemical bond, -O-, -NHC(O)O-, -OC(O)NH-, -N(CH3)C(O)O- or -C(O)O-;

[0835] L is selected from B10. The oligonucleotide of any one of technical solutions B6-B9, wherein the compound of formula (Ia) is selected from the following structures:

[0836] in,

[0837] It represents a chemical bond to the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide;

[0838] X1 is a chemical bond connecting to the hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose of another adjacent nucleotide;

[0839] X3 is selected from O or S;

[0840] Preferably, the compound of formula (Ia) is selected from:

[0841] It represents the chemical bond connecting to the hydroxyl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide;

[0842] X1 is a chemical bond connecting to the hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose of another adjacent nucleotide;

[0843] X3 is selected from O or S.

[0844] B11. The oligonucleotide of any one of technical solutions B6-B10, which has 14 to 30 nucleotides.

[0845] B12. The oligonucleotide of any one of technical solutions B6-B11, which contains one or more compounds of formula (Ia), or pharmaceutically acceptable salts, tautomers or stereoisomers thereof, inside the oligonucleotide.

[0846] B13. A double-stranded RNA having a sense strand and an antisense strand, each strand having 14 to 30 nucleotides, the antisense strand comprising a sequence that is sufficiently complementary to the sense strand and the target mRNA, wherein the sense strand and / or antisense strand comprises one or more compounds of formula (II) described in any one of technical solutions B1-B5 or compounds of formula (Ia) described in any one of technical solutions B6-B10, or pharmaceutically acceptable salts, tautomers or stereoisomers thereof.

[0847] B14. The double-stranded RNA described in technical solution B13, wherein the positive strand comprises one or more compounds of formula (Ia) described in any one of technical solutions B6-B10, or pharmaceutically acceptable salts, tautomers or stereoisomers thereof, inside the oligonucleotide.

[0848] B15. The double-stranded RNA described in technical solution B13, wherein the antisense strand comprises a compound of formula (II) described in any one of technical solutions B1-B5 at the 5' end, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[0849] B16. The double-stranded RNA described in technical solution B13, wherein the antisense strand comprises one or more compounds of formula (Ia) described in any one of technical solutions B6 to B10, or pharmaceutically acceptable salts, tautomers or stereoisomers thereof, inside the oligonucleotide.

[0850] B17. The double-stranded RNA of any one of technical solutions B13-B16, which is selected from small interfering RNA (siRNA) and short hairpin RNA (shRNA).

[0851] B18. A cell containing the double-stranded RNA according to any one of technical solutions B14 to B17.

[0852] B19. A pharmaceutical composition comprising the double-stranded RNA according to any one of technical solutions B14 to B17, or the cell according to technical solution B18, and optionally a pharmaceutically acceptable carrier or excipient.

[0853] B20. A kit comprising the double-stranded RNA according to any one of technical solutions B14 to B17, the cell according to technical solution B18, or the pharmaceutical composition according to technical solution B19.

[0854] B21. A compound of formula (IIb), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0855] in,

[0856] Each R T1 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0857] Each R T2 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0858] m is 0, 1, 2, 3, 4 or 5;

[0859] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0860] R X1 Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0861] L is -Ar-(CH2)-O-, wherein CH2 may be optionally substituted by 1 or 2 R#, R# being selected from H, D, C1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0862] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0863] R*selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated;

[0864] PG is selected from protecting groups.

[0865] B22. The compound of technical solution B21, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein

[0866] Each R T1 Independently selected from H, D, C 1-6 Alkyl, C 1-6 a haloalkyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated;

[0867] Each R T2 Independently selected from H, D, C 1-6 Alkyl, C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0868] m is 0, 1, 2, or 3;

[0869] X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -;

[0870] R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0871] L is -Ar-(CH2)-O-, wherein CH2 may be optionally substituted by 1 or 2 R#, R# being selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0872] Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*;

[0873] R*selected from C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated;

[0874] PG is selected from protecting groups.

[0875] B23. The compound of technical solution B21 or B22, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein PG is selected from 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 (Tro c), 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), 4,4'-dimethoxytrityl, -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH, preferably -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH.

[0876] B24. The compound of any one of technical solutions B21-B23, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the compound of formula (IIb) is selected from:

[0877] in,

[0878] PG is -P(OCH2CH2CN)(N(iPr)2) or H.

[0879] B25. The oligonucleotide of any one of technical solutions B1-B3 and B6-B8 or the compound of formula (IIb) of technical solution B21 or B22, wherein the chain containing GalNAc is a conjugated group represented by formula (X'):

[0880] in,

[0881] Indicates the location of attachment to a biomolecule;

[0882] Q is independently H,

[0883] Wherein L1 is a chemical bond, -CH2-, -CH2CH2-, -C(O)-, -CH2O-, -CH2O-CH2CH2O- or -NHC(O)-(CH2NHC(O)) a -;

[0884] L2 is a chemical bond or -CH2CH2C(O)-;

[0885] L3 is a chemical bond, -(NHCH2CH2) b -、-(NHCH2CH2CH2) b - or -C(O)CH2-;

[0886] L4 is -(OCH2CH2) c -、-(OCH2CH2CH2) c -、-(OCH2CH2CH2CH2) c -、-(OCH2CH2CH2CH2CH2) c -or-NHC(O)-(CH2) d -;

[0887] Where a = 0, 1, 2 or 3;

[0888] b = 1, 2, 3, 4, or 5;

[0889] c = 1, 2, 3, 4, or 5;

[0890] d = 1, 2, 3, 4, 5, 6, 7, or 8;

[0891] A is a chemical bond, -CH2O- or -NHC(O)-;

[0892] A' is a chemical bond, -C(O)NH-, -NHC(O)- or -O(CH2CH2O) e -;

[0893] Where e is 1, 2, 3, 4 or 5;

[0894] B is a chemical bond, -CH2-, -C(O)-, -M-, -CH2-M- or -C(O)-M-;

[0895] Where M is

[0896] R1 and R2 together form -CH2CH2O- or -CH2CH(R)-O-, and R3 is H;

[0897] Or R1 and R3 together form -C 1-2 Alkylene-, and R2 is H;

[0898] wherein R is -OR', -CH2OR' or -CH2CH2OR', wherein R' is H, a hydroxyl protecting group or a solid phase support, and the hydroxyl protecting group is preferably -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytrityl;

[0899] m1 = 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0900] n1=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. Example

[0901] The following examples are intended to illustrate the present invention but are not intended to limit the scope of the present invention.

[0902] abbreviation

[0903] Example 1 Preparation of DN0110

[0904] 1. Preparation of Compound 1c

[0905] Compound 1a (40 g, 425 mmol, 37.3 mL) was dissolved in CH₃SO₃H (240 mL), and compound 1b (72.7 g, 637 mmol, 77.8 mL) was added. The mixture was stirred at 70°C for 96 hours. The reaction mixture was cooled to 0°C and quenched with water (500 mL). The mixture was then extracted with ethyl acetate (500 mL x 2). The organic phase was washed with brine (100 mL x 2), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The organic phase was purified on a silica gel column (petroleum ether:ethyl acetate = 10:1) to afford compound 1c (43.1 g, 57%).

[0906] 1H NMR (400MHz CDCl3) δ7.63(dd,J=1.6Hz,3.6Hz,1H),7.24-7.29(m,1H),7.14-7.18(m,1H),7.63(dd,J=1.2Hz,8.0Hz,1H),2.64(s,2H),1.37(s,6H).

[0907] m / z: ES+[M+H]+177.0.

[0908] 2. Preparation of Compound 1d

[0909] Lithium aluminum hydride (8.61 g, 227 mmol) was suspended in tetrahydrofuran (350 mL) and replaced with nitrogen three times. A solution of compound 1c (20.0 g, 113 mmol) dissolved in tetrahydrofuran (140 mL) was then added dropwise at 0°C. The mixture was then stirred at 0°C for 1.5 hours. Saturated ammonium chloride solution (150 ml) was slowly added to the reaction mixture. The reaction mixture was filtered through celite. The mixture was diluted with ethyl acetate (200 x 2), washed twice with brine (300 ml x 2), and the organic phase was dried over anhydrous sodium sulfate, filtered, and distilled under reduced pressure to give compound 4 (20.4 g, 97%).

[0910] 1 H NMR(400MHz CDCl3)δ7.22(dd,J=0.8Hz,4.0Hz,1H),7.04-7.10(m,1H),6.82-6.09(m,1H),6.64(dd,J=0.8 Hz,4.0Hz,1H),6.17-6.31(m,1H),3.53(t,J=4.0Hz,2H),2.23(t,J=4.0Hz,2H),1.42(6H,s).

[0911] 3. Preparation of Compound 1f

[0912] A mixture of compound 1d (15.0 g, 83.2 mmol) and cesium carbonate (81.3 g, 249 mmol) in DMF (150 mL) was purged with nitrogen three times. Compound 1e (20.0 g, 124 mmol, 17.4 mL) was then added, and the reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was quenched with water (100 mL) and extracted with ethyl acetate (100 mL x 2). The combined organic layers were washed with brine (200 mL x 2), dried over anhydrous sodium sulfate, and filtered. The organic phase was concentrated under reduced pressure and purified on a silica gel column (petroleum ether:ethyl acetate = 3:1) to afford compound 1f (10.8 g, 53.9%).

[0913] 1 H NMR (400MHz CDCl3) δ7.17-7.28(m,2H),6.88-6.98(m,2H),5.2(s,2H),3.45(t,J=4.0Hz,2H),2.31(s,3H),2.20(t,J=7.2Hz,2H),1.3(s,6H).

[0914] 4. Preparation of Compound 1g

[0915] Compound 1f (10.8 g, 44.9 mmol) was dissolved in pyridine (100 mL), and acetic anhydride (13.7 g, 134 mmol, 12.6 mL) and DMAP (548 mg, 4.49 mmol) were added sequentially. The reaction mixture was stirred at 25°C for 2 hours. Ethanol (50 mL) was added to the solution, and the mixture was concentrated under reduced pressure. The resulting residue was dissolved in ethyl acetate (100 mL) and washed with saturated sodium bicarbonate solution (120 mL) and brine (120 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield compound 1g (12.0 g, 94.5%).

[0916] 1 H NMR(400MHz CDCl3)δ7.16-7.26(m,2H),6.88-6.97(m,2H),5.19(s,2H),3.88(t,J=4. 0Hz, 2H), 2.31 (s, 3H), 2.25 (t, J = 4.0Hz, 1.6H), 1.89 (s, 3H), 1.43 (s, 6H).

[0917] 5. Preparation of Compound 1h

[0918] Under a nitrogen atmosphere, SO2Cl2 (2.22 g, 16.4 mmol, 1.65 mL) was slowly added to a solution of compound 1g (5.00 g, 17.7 mmol) in dichloromethane (50 mL). The mixture was stirred at 25°C for 2 hours. The reaction mixture was dried under vacuum. The mixture was dissolved in dichloromethane (20 mL) and DMF (12 mL). p-TsSK (6.01 g, 26.5 mmol) was added to the solution and stirred at 25°C for 1 hour. Tert-butyl mercaptan (3.19 g, 35.4 mmol, 3.99 mL) was then added, and the reaction mixture was stirred at 25°C under a nitrogen atmosphere for 18 hours. The reaction solution was diluted with ethyl acetate (120 mL), washed with water (100 mL x 3), and the organic phase was dried over anhydrous sodium sulfate and filtered. The organic phase was concentrated under reduced pressure and purified on a silica gel column (petroleum ether:ethyl acetate = 10:1) to give compound 1h (4.42 g, 69.7%).

[0919] 1 H NMR(400MHz CDCl3)δ7.14-7.26(m,2H),6.87-6.98(m,2H),5.34(m,2H),3.85(t,J=4.0Hz,2H),2.25(t,J=4.0Hz,2H),1.89(s,3H),1.42(s,6H),1.37(s,9H).

[0920] 6. Preparation of Compound 1i

[0921] Compound 1h (4.4 g, 12.3 mmol) and potassium carbonate (3.41 g, 24.6 mmol) were dissolved in methanol (40 mL) and stirred at 25°C for 2 hours. The reaction mixture was diluted with ethyl acetate (50 mL) and washed twice with brine (100 mL x 2). The organic layer was dried over anhydrous sodium sulfate and filtered. The organic phase was concentrated under reduced pressure and purified on a silica gel column (petroleum ether:ethyl acetate = 5:1) to afford compound 1i (1.19 g, 30.8%).

[0922] 1 H NMR (400MHz CDCl3) δ7.18-7.27(m,2H),6.89-6.99(m,2H),5.35(s,2H),3.43(t,J=2.0Hz,2H),2.20(t,J=4.0Hz,2H),1.42(s,6H),1.38(s,9H).

[0923] 7. Preparation of Compound DN0110

[0924] Compound 1i (1.00 g, 3.18 mmol) and DCI (187 mg, 1.59 mmol) were dissolved in dichloromethane (10 mL), and compound 1j (1.92 g, 6.36 mmol, 2.02 mL) was added. The reaction mixture was stirred at 25°C for 1 hour. Triethylamine (0.5 mL) and triethylamine-basified silica gel were added to the reaction solution, which was then concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether (1% triethylamine) / ethyl acetate = 98 / 2) to yield compound DN0110 (1.03 g, 62.9%).

[0925] 1 H NMR(400MHz CDCl3)δ7.16-7.27(m,2H),6.89-6.99(m,2H),5.33(m,2H),3.67-3.80(m,2H),3.47-3.59(m,2H),3.30-3.45(m,2H ),2.58(t,J=4.0Hz,2H),2.17-2.25(m,2H),1.41(s,6H),1.37(s,9H),1.15(t,J=4.0Hz,6H),1.09(t,J=4.0Hz,6H)

[0926] 31 P NMR (400MHz CDCl3) δ 146.50.

[0927] Example 2 Preparation of DN0111

[0928] The synthesis steps of Example 2 are the same as those of Example 1, wherein the compound tert-butyl mercaptan is replaced by the compound isopropyl mercaptan.

[0929] 1 H NMR (400MHz, CDCl3) δ7.09-7.17(m,2H),6.82-6.91(m,2H),5.21(s,2H),3.61-3.70(m,2H),3.440-3.57(m,4H),3.18-3.27(m,1H),2. 99-3.05(m,1H),2.51(t,J=4Hz,1H),2.111-2.21(m,2H),1.33-1.35(m,6H),1.24-1.26(m,6H),1.18-1.22(m,6H),1.05-1.08(m,6H).

[0930] 31 P NMR (400MHz CDCl3) δ 146.52.

[0931] Example 3 Preparation of Compound AE1

[0932] 1. Preparation of Compound 3

[0933] Compound 1 (13.0 g, 104 mmol) and cesium carbonate (102 g, 314 mmol) were dissolved in DMF (130 mL). After nitrogen substitution three times, compound 2 (15.2 g, 157 mmol, 13.2 mL) was added to the previous solution and reacted at 25°C for 2 hours. TLC (petroleum ether:ethyl acetate = 3:1) showed the disappearance of the starting material and the formation of new spots. The reaction solution was diluted with ethyl acetate and extracted (50 mL x 3). The organic phase was washed sequentially with saturated sodium bicarbonate (50 mL) and saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, and dried over a silica gel column (petroleum ether:ethyl acetate = 5:1-3:1) to obtain compound 3 (10.0 g, 54.3 mmol, 51.82%) as a yellow oil.

[0934] 1 H NMR (400MHz, CDCl3) δ7.32-7.27(m,2H),6.96-6.94(m,2H),5.20(s,2H),4.62(s,2H),2.25(s,3H).

[0935] 2. Preparation of Compound 4

[0936] Compound 3 (10 g, 54.27 mmol) was dissolved in pyridine (100 mL), and acetic anhydride (6.65 g, 65.13 mmol, 6.12 mL) and 4-dimethylaminopyridine were added. The mixture was reacted at 25°C for 3 hours. TLC (petroleum ether:ethyl acetate = 3:1) showed the disappearance of the starting material and the formation of new spots. The reaction solution was quenched with ethanol (10 mL), the solvent was dried by spin-drying, and then extracted with ethyl acetate (100 mL x 3). The organic phase was washed sequentially with saturated sodium bicarbonate (50 mL) and saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, and dried by passing through a silica gel column (petroleum ether:ethyl acetate = 5:1-3:1) to afford compound 4 (4 g, 17.32 mmol, 31.92%) as a colorless oil.

[0937] 1 H NMR (400MHz, DMSO) δ7.31 (d, J = 8.78Hz, 2H), 6.99 (d, J = 8.53Hz, 2H), 5.27 (s, 2H), 5.00 (s, 2H), 2.16 (s, 3H), 2.03 (s, 3H).

[0938] 3. Preparation of Compound 5

[0939] Under a nitrogen atmosphere, compound 4 (5.0 g, 22.1 mmol) was dissolved in CHCl (50 mL). SOCl (2.98 g, 22.1 mmol, 2.21 mL) dissolved in CHCl (5 mL) was slowly added. The mixture was stirred at 25°C for 2 hours. The reaction mixture was dried under vacuum. The mixture was dissolved in CHCl (20 mL) and DMF (12 mL). P-TsSK (7.50 g, 33.1 mmol) was added to the solution, and the mixture was stirred at 25°C for 1 hour. Tert-butyl mercaptan (3.99 g, 44.2 mmol, 4.98 mL) was then added, and the reaction mixture was stirred at 25°C under a nitrogen atmosphere for 18 hours. TLC (petroleum ether:ethyl acetate = 10:1) showed the formation of new spots. The reaction solution was diluted with ethyl acetate (60 mL) and washed with water (50 mL x 3). The organic layer was dried over Na2SO4 and filtered. The organic phase was concentrated under reduced pressure and purified using a normal phase column (petroleum ether:ethyl acetate = 10:1) to give compound 5 (4.90 g, 16.3 mmol, 73.81%) as a yellow oil. 1 H NMR (400MHz, CD3OD) δ7.32-7.27(m,2H),6.96-6.94(m,2H),5.20(s,2H),4.62(s,2H),2.25(s,3H).

[0940] 4. Preparation of Compound 6

[0941] Compound 5 (4.90 g, 16.3 mmol) and K2CO3 (4.51 g, 32.6 mmol) were dissolved in MeOH (50 mL) and stirred at 25°C for 2 hours. TLC (petroleum ether:ethyl acetate = 3:1) showed the formation of a new spot. The mixture was concentrated under reduced pressure, then diluted with ethyl acetate (20 mL) and washed twice with brine (50 mL x 2). The organic layer was dried over Na2SO4, concentrated under reduced pressure, and purified using a normal phase column (petroleum ether:ethyl acetate = 3:1) to afford Compound 6 (2.1 g, 7.09 mmol, 43.50%) as a yellow oil.

[0942] 1 H NMR: (400MHz, CD3OD) δ7.31-7.29(m,2H),6.95–6.91(m,2H),5.33(s,2H),4.50(s,2H),1.32(s,9H).

[0943] 5. Preparation of Compound AE1

[0944] Compound 6 (800 mg, 3.10 mmol) and DCI (183 mg, 1.55 mmol) were dissolved in CH2Cl2 (8 mL), and compound 7 (1.87 g, 6.19 mmol) was added. The reaction mixture was stirred at 25°C for 1 hour. TLC (petroleum ether (1% triethylamine) / ethyl acetate = 10 / 1) indicated complete reaction of the starting materials. Triethylamine (0.5 mL) and triethylamine-basified silica gel were added to the reaction solution, which was then concentrated under reduced pressure. The crude product was separated by normal phase column chromatography (petroleum ether (1% triethylamine) / ethyl acetate = 98 / 2) to afford AE1 (560 mg, 1.22 mmol, 39.44%) as a colorless oil.

[0945] 1 H NMR (400MHz, CDCl3) δ7.29-7.31(m,2H),6.91-6.93(m,2H),5.29(s,2H),4.59-4.74(m,2H),3 .81-3.88(m,2H),3.61-3.70(m,2H),2.63(t,J=6.8Hz,2H),1.35(s,9H),1.18-1.22(m,12H).

[0946] Example 4 Preparation of Compound AE2

[0947] 1. Preparation of Compound 2

[0948] Compound 1 (5.00 g, 40.9 mmol) was dissolved in THF (40 mL). Isopropylmagnesium chloride (i-PrMgCl) (2 M, 81.9 mL) was added at 0°C, and the reaction was incubated at 0°C for 3 hours. TLC (petroleum ether:ethyl acetate = 3:1) revealed the formation of new spots. The reaction mixture was cooled to 0°C and quenched with ammonium chloride (80 mL), followed by extraction with ethyl acetate (100 mL x 2). The organic phase was washed with brine (100 mL x 2), dried over anhydrous Na2SO4, and filtered to yield compound 2 (6.70 g, crude) as a brown solid.

[0949] 1 H NMR (400MHz, CD3OD) δ7.11(d,J=8.4Hz,2H),6.73(d,J=8.4Hz,2H),4.15(d,J =7.2Hz,1H),1.83-1.89(m,1H),0.98(d,J=7.6Hz,3H),0.72(d,J=7.6Hz,3H).

[0950] 2. Preparation of Compound 4

[0951] Compound 2 (2 g, 12.0 mmol) and cesium carbonate (11.8 g, 36.1 mmol) were dissolved in DMF (20 mL) and the atmosphere was replaced with nitrogen three times. Compound 3 (1.74 g, 18.1 mmol, 1.51 mL) was then added. The mixture was stirred at 25°C for 1 hour. TLC (petroleum ether:ethyl acetate = 3:1) showed the formation of new spots. The mixture was diluted with ethyl acetate (40 mL) and washed twice with brine (30 mL). The organic phase was dried over Na2SO4, filtered, and dried to afford Compound 4 (2.8 g, crude) as a yellow oil.

[0952] 1 H NMR (400MHz, CD3OD) δ7.23(d,J=8.8Hz,2H),6.93(d,J=8.8Hz,2H),5.18(s,2H),4.22(d,J =7.2Hz,1H),2.21(s,3H),1.83-1.93(m,1H),0.99(d,J=6.4Hz,3H),0.74(d,J=6.8Hz,3H).

[0953] 3. Preparation of Compound 5

[0954] Compound 4 (2.70 g, 11.9 mmol) was dissolved in pyridine (25 mL), and acetic anhydride (3.65 g, 35.8 mmol, 3.36 mL) and DMAP (146 mg, 1.19 mmol) were added sequentially. The reaction mixture was stirred at 25°C for 2 hours. TLC (petroleum ether:ethyl acetate = 3:1) showed the formation of new spots. The mixture was evaporated in vacuo. The residue was dissolved in ethyl acetate (30 mL), and the resulting solution was washed with NaHCO₃ solution (120 mL) and brine (120 mL). The organic layer was dried over Na₂SO₄, filtered, and concentrated in vacuo to afford compound 5 (2.87 g, 10.69 mmol, 89.65%) as a yellow oil.

[0955] 1 H NMR(400MHz,CD3OD)δ7.22(d,J=8.8Hz,2H),6.94(d,J=8.8Hz,2H),5.18(s,2H),2.20 (s,3H),2.07-2.13(m,1H),2.02(s,3H),1.97(d,J=8.0Hz,3H),0.78(d,J=6.8Hz,3H).

[0956] 4. Preparation of Compound 6

[0957] To a solution of compound 5 (2.8 g, 10.4 mmol) in CH2Cl2 (30 mL) under a nitrogen atmosphere, SO2Cl2 (1.31 g, 9.70 mmol, 970 μL) dissolved in CH2Cl2 (5 mL) was slowly added. The mixture was stirred at 25°C for 2 hours. The reaction mixture was dried under vacuum. The mixture was dissolved in CH2Cl2 (10 mL) and DMF (6 mL). P-TsSK (3.54 g, 15.7 mmol) was added to the solution, and the mixture was stirred at 25°C for 1 hour. Tert-butyl mercaptan (941 mg, 10.4 mmol, 1.17 mL) was then added, and the reaction mixture was stirred at 25°C under a nitrogen atmosphere for 18 hours. TLC (petroleum ether:ethyl acetate = 10:1) showed the formation of a new spot. The reaction solution was diluted with ethyl acetate (60 mL), washed with water (50 mL x 3), and the organic layer was dried over Na2SO4 and filtered. The organic phase was concentrated under reduced pressure and purified using a normal phase column (petroleum ether: ethyl acetate = 10:1) to give compound 6 (2.10 g, 6.13 mmol, 58.7%) as a yellow oil.

[0958] 1 H NMR (400MHz, CD3OD) δ7.23-7.26(m,2H),6.91-6.94(m,2H),5.37(d,J=8.0Hz,1H),5.32(s,2H ),2.06-2.12(m,1H),2.04(s,3H),1.32(s,9H),0.97(d,J=6.4Hz,3H),0.78(d,J=6.4Hz,3H).

[0959] 5. Preparation of Compound 7

[0960] Compound 6 (2.10 g, 6.13 mmol) and K2CO3 (1.69 g, 12.3 mmol) were dissolved in MeOH (20 mL) and stirred at 25°C for 2 hours. TLC (petroleum ether:ethyl acetate = 10:1) showed the formation of a new spot. The reaction mixture was diluted with ethyl acetate (20 mL) and washed twice with brine (100 mL x 2). The organic layer was dried over Na2SO4, concentrated under reduced pressure, and purified using a normal phase column (petroleum ether:ethyl acetate = 10:1) to afford compound 7 (1.2 g, 3.88 mmol, 63.31%) as a yellow oil.

[0961] 1H NMR (400MHz, CD3OD) δ7.23-7.26(m,2H),6.90-6.93(m,2H),5.33(s,2H),4.22(d,J=7. 2Hz,1H),1.85-1.93(m,1H),1.32(s,9H),0.98(d,J=6.8Hz,3H),0.74(d,J=6.8Hz,3H).

[0962] 6. Preparation of Compound AE2

[0963] Compound 7 (500 mg, 1.66 mmol) and DCI (98.3 mg, 832 μmol) were dissolved in CH2Cl2 (10 mL) and compound 8 (1.00 g, 3.33 mmol, 1.06 mL) was added. The reaction mixture was stirred at 25°C for 1 hour. TLC (petroleum ether (1% triethylamine) / ethyl acetate = 10 / 1) indicated complete reaction of the starting materials. Triethylamine (0.5 mL) and silica gel basified with triethylamine were added to the reaction solution, which was then concentrated under reduced pressure. The crude product was separated by normal phase column chromatography (petroleum ether (1% triethylamine) / ethyl acetate = 98 / 2) to afford compound AE2 (310 mg, 619 μmol) as a colorless oil.

[0964] 1 H NMR (400MHz, CDCl3) δ7.22(d,J=8.4Hz,2H),6.86-6.91(m,2H),5.28(d,J=5.6Hz,2H),4.40-4.46(m,1H),3.62-3.89(m,2H),3.48-3.56(m, 2H),2.64(t,J=6.4Hz,1H),2.33(t,J=6.4Hz,1H),194-1.99(m,1H),1.35(s,9H),1.12-1.23(m,9H),0.91-0.97(m,6H),0.78-0.80(m,3H).

[0965] Example 5 Preparation of Compound AE3

[0966] 1. Preparation of Compound 124-1

[0967] To a solution of benzo[b]thiophene-2-boronic acid (10.0 g, 56.2 mmol) in ethanol (50 mL) was added 30% hydrogen peroxide (20 mL) dropwise over 30 minutes. The reaction was then stirred for 16 hours. TLC indicated the reaction was complete. The reaction mixture was then carefully washed three times with water and extracted twice with dichloromethane. The organic phase was dried and concentrated to obtain the crude product, which was then purified on a silica gel column (PE / EA = 5:1) to afford compound 124-1 (7.8 g, 92.1%) as a pale yellow solid.

[0968] 1 H NMR (400MHz, CDCl3) δ7.50–7.08 (m, 4H), 3.99 (s, 2H).

[0969] 2. Preparation of Compound 124-2

[0970] A solution of LiAlH4 (3.9 g, 102.7 mmol) in THF (80 mL) was cooled to 0°C, followed by dissolving compound 124-1 in THF (30 mL) over 1 hour and adding it to the reaction mixture. The reaction was stirred for 16 hours. TLC showed that the reaction was complete, and the reaction mixture was carefully quenched with saturated Na2SO4 solution, followed by addition of 1M HCl solution until the reaction mixture clarified. The organic phase was dried and concentrated to obtain the crude product, which was then purified by silica gel column (PE / EA = 2:1) to obtain compound 124-2 (6.0 g, 75.2%) as a colorless oil.

[0971] 1 H NMR (400MHz, CDCl3) δ7.23(dd,J=7.3,1.8Hz,1H),7.12(dd,J=7.2,2.0Hz,1H),7.04(pd,J =7.3,1.8Hz,2H),3.81(t,J=6.6Hz,2H),3.34(s,1H),2.90(t,J=6.6Hz,2H).

[0972] 3. Preparation of Compound 124-3

[0973] To a solution of 2,2'-disulfide dipyridine (PySSPy, 21.4 g, 97.1 mmol) and acetic acid (1.2 mL) in methanol (100 mL) was added a solution of compound 124-2 (6.0 g, 38.9 mmol) in methanol (20 mL). The reaction was stirred for 16 hours. TLC indicated the reaction was complete, and the reaction was then concentrated to obtain the crude product, which was purified by silica gel column chromatography to obtain compound 124-3 (5.7 g, 56.0%) as a colorless oil.

[0974] 1 H NMR (400MHz, CDCl3) δ8.50 (dt, J=4.9, 1.4Hz, 1H), 7.64 (ddt, J=8.2, 7.2, 3.2Hz, 3H) ,7.27–7.17(m,3H),7.15–7.11(m,1H),3.97(t,J=6.6Hz,2H),3.18(t,J=6.6Hz,2H).

[0975] 4. Preparation of Compound 124-4

[0976] To a solution of compound 124-3 (5.7 g, 21.7 mmol) in dichloromethane (60 mL) was added methyl trifluoromethanesulfonate (MeOTf, 7.1 g, 43.4 mmol) dropwise, and the reaction was stirred for 15 minutes. Tert-butyl mercaptan (2.0 g, 21.7 mmol) and DIPEA (7.6 mL, 43.4 mmol) were added to the reaction mixture. The reaction was then stirred for 30 minutes. TLC showed that the reaction was complete, and the reaction was directly concentrated to obtain a crude product, which was purified by silica gel column (PE / EA = 6:1) to obtain compound 124-4 (2.9 g, 55.6%) as a colorless oil.

[0977] 1 H NMR (400MHz, CDCl3) δ7.77(dd,J=7.9,1.1Hz,1H),7.19–7.05(m,3H),3.84(t,J=6.8Hz,2H),3.05(t,J=6.7Hz,2H),1.23(s,9H).

[0978] 5. Preparation of Compound 124-5

[0979] Phosphorous acid (9.8 g, 120.0 mmol) was co-evaporated three times using anhydrous pyridine and then redissolved in pyridine (60 mL). Compound 124-4 (2.9 g, 12.0 mmol) was added to the reaction mixture, and the reaction was then cooled to 0°C and stirred for 10 minutes. Trimethylacetyl chloride (7.2 g, 60.0 mmol) was added to the reaction mixture and stirred for 3 hours. The reaction was quenched with triethylammonium bicarbonate buffer (30 mL, 1 M) and diluted twice with ethyl acetate. The organic phases were then combined and concentrated to give the crude product, which was purified by silica gel column (MeOH:DCM=10:1, 1% TEA) to give compound 124-5 (860 mg, 59.7%) as a colorless oil.

[0980] 6. Preparation of Compound 124-7

[0981] A solution of compound 124-5 (860 mg, 2.1 mmol) and compound 124-6 (465.0 mg, 1.8 mmol) was co-evaporated three times using anhydrous pyridine and redissolved in pyridine (15 mL). The reaction was cooled to -15 ° C and trimethylacetyl chloride (0.5 mL, 3.6 mmol) was added dropwise and stirred at -15 ° C for 2 hours. The reaction was diluted with dichloromethane and quenched with saturated NH4Cl solution. The organic phases were combined and concentrated to obtain a crude product, which was purified by silica gel column (MeOH: DCM = 5: 1, 1% acetic acid) to obtain compound 124-7 (650 mg, 56.2%) as a white solid.

[0982] 7. Preparation of Compound 124-8

[0983] To a solution of compound 124-7 (650 mg, 1.2 mmol) in dichloromethane and carbon tetrachloride (v / v = 1:1, 16 mL) was added benzylamine (0.6 mL, 5.5 mmol), and the reaction was stirred for 16 hours. TLC indicated the reaction was complete, and the reaction was then concentrated to obtain the crude product, which was purified on a silica gel column (MeOH:DCM = 8:1) to obtain compound 124-8 (580 mg, 75.0%) as a white solid.

[0984] 8. Preparation of Compound AE3

[0985] To a solution of compound 124-8 (580 mg, 0.9 mmol) in dichloromethane (8 mL) were added compound 124-9 (352 mg, 1.2 mmol) and DCI (159.6 mg, 1.4 mmol), and the reaction was stirred for 2 hours. TLC indicated the reaction was complete, and the mixture was washed with water and extracted with dichloromethane. The organic phases were combined and concentrated to yield the crude product, which was then purified on a silica gel column (PE:EA = 6:1, 1% pyridine) to afford compound AE3 (640 mg, 85.2%) as a white solid.

[0986] 1H NMR (400MHz, CDCl3) δ7.91(s,1H),7.69(dd,J=8.2,5.0Hz,1H),7.58(dd,J=8.2,3.1Hz,1H),7.35–7.27(m,3H),7.25–7.15(m,2H ),6.97–6.91(m,1H),6.87(d,J=5.0Hz,1H),6.00–5.92(m,1H),5.64(dd,J=8.2,6.6Hz,1H),5.56(dd,J=8.2,6.7Hz,1H),5.31–5. 29(m,2H),4.00(dtd,J=12.9,6.8,6.4,2.8Hz,3H),3.88–3.74(m,5H),3.69(tt,J=12.5,6.3Hz,2H),3.62(tt,J=12.5,6.3Hz,3H) ,3.49(d,J=1.0Hz,1H),3.45(d,J=6.0Hz,2H),2.29(t,J=7.2Hz,3H),1.34(d,J=1.2Hz,9H),1.19(ddd,J=15.2,6.9,3.6Hz,12H). 31 P NMR (162MHz, CDCl3) δ151.07,150.78,150.28,150.23,8.92,8.73,8.42,8.31.

[0987] Example 6 Preparation of Compound AE4

[0988] 1. Preparation of Compound 125-4

[0989] To a solution of phosphonic acid (5.53 g, 67.45 mmol) and compound 125-3 (6.70 g, 13.49 mmol) in anhydrous pyridine (Py, 50 mL) was added trimethylacetyl chloride (4.07 mL, 33.73 mmol) and stirred, followed by TLC analysis of the reaction. After 3 hours, the reaction was quenched by the careful addition of 2 M triethylamine bicarbonate buffer (TEAB, 20 mL). After carbon dioxide evolution ceased, the reaction mixture was evaporated to dryness and the residue was partitioned between dichloromethane and 0.5 M TEAB. The organic layer was evaporated and the residue was purified on a silica gel column to obtain compound 125-4 (8.40 g, 94%).

[0990] 2. Preparation of Compound 125-5

[0991] To a stirred solution of compound 125-4 (2.5 g, 3.8 mmol) and compound 1i (1.2 g, 3.8 mmol) in anhydrous pyridine (10 mL) was added trimethylacetyl chloride (1.4 g, 11.3 mmol). After TLC analysis indicated completion of the reaction, 2 M TEAB (1 mL) was added. The reaction mixture was concentrated and the residue was partitioned between dichloromethane and 0.5 M TEAB. The organic layer was collected, treated with Na2SO4, and evaporated. Purification on a silica gel column afforded a diisomeric mixture of compound 125-5 (crude, 1.7 g, 53%).

[0992] 3. Preparation of Compound 125-6

[0993] To a solution of compound 125-5 (1.7 g, 2.0 mmol) in dichloromethane and carbon tetrachloride (v / v = 1:1, 4 ml) was added benzylamine (0.3 g, 3 mmol) dropwise and the resulting mixture was stirred for 3 hours. The volatiles were removed in vacuo to obtain a residue, which was then purified on a flash silica gel column to obtain compound 125-6 (1.5 g, 79%) as a white solid.

[0994] 4. Preparation of Compound 125-7

[0995] Compound 125-6 was dissolved in a 1M solution of TBAF in THF (0.5 mL) and stirred overnight. The reaction mixture was then separated between water and diethyl ether. The organic layer was collected, dried over Na2SO4, and evaporated. It was then purified on a silica gel column to obtain a non-corresponding isomeric mixture of compound 125-7 (crude, 1.1 g, 97%).

[0996] 5. Preparation of Compound AE4

[0997] Compound 125-7 (0.60 g, 0.83 mmol) and 5-(ethylthio)-1H-tetrazole (ETT, 0.08 g, 0.66 mmol) in anhydrous dichloromethane (5 mL) were added compound 125-8 (0.37 g, 1.24 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was filtered, concentrated, and loaded onto a silica gel column to obtain compound AE4 (0.50 g, 65%) as a white foam.

[0998] 1H NMR (400MHz, CDCl3) δ7.91(s,1H),7.69(dd,J=8.2,5.0Hz,1H),7.58(dd,J=8.2,3.1Hz,1H),7.35–7.27(m,3H),7.25–7.15(m,4H),6.97–6.91(m ,1H),6.87(d,J=5.0Hz,1H),6.00–5.92(m,1H),5.64(dd,J=8.2,6.6Hz,1H),5.56(dd,J=8.2,6.7Hz,1H),5.31–5.29(m,2H),4.41–4.25(m,3H), 4.24–4.13(m,3H),4.00(dtd,J=12.9,6.8,6.4,2.8Hz,3H),3.88–3.74( m,5H),3.62(tt,J=12.5,6.3Hz,3H),3.49(d,J=1.0Hz,1H),3.45(d,J=6 .0Hz,2H),2.98–2.72(m,3H),2.66–2.54(m,3H),2.29(t,J=7.2Hz,3H), 1.40(s,6H),1.34(d,J=1.2Hz,9H),1.19(ddd,J=15.2,6.9,3.6Hz,12H). 31 P NMR (162MHz, CDCl3) δ151.07,150.78,150.28,150.23,8.92,8.73,8.42,8.31.

[0999] Example 7 Preparation of Compound AE5

[1000] 1. Preparation of Compound 3b

[1001] LiAlH4 (57.08 mg, 142.69 mmol) was added to THF (150 mL). The reaction mixture was cooled to 0°C, followed by the addition of compound 3a (10.00 g, 64.86 mmol). The reaction was allowed to proceed at 20°C for 14 hours and then at 25°C for 24 hours. The reaction mixture was cooled to 0°C and quenched with 2N hydrochloric acid. The aqueous phase was extracted twice with 100 mL of ethyl acetate. The organic phase was filtered and dried to afford compound 3b (8.90 g, 63.48 mmol, 97.9%).

[1002] 1 H NMR (400MHz, CD3OD) δ7.18-7.28 (m, 4H), 4.54 (s, 2H).

[1003] 2. Preparation of Compound 3c

[1004] Compound 3b (5.00 g, 35.7 mmol) was dissolved in EtOH (100 mL), and tert-butyl mercaptan (36.95 mL, 340.1 mmol) was added. A solution of iodine (6.79 g, 26.7 mmol) in EtOH (100 mL) was added dropwise at 0°C, followed by a 20°C reaction for 2 hours. After completion of the reaction, the pH was adjusted to 7 with saturated sodium bicarbonate. The ethanol was partially removed by vortexing, and the aqueous phase was extracted with ethyl acetate (300 mL x 3). The organic phase was filtered and dried to obtain the crude product, which was then purified by column chromatography (petroleum ether / ethyl acetate) to afford compound 3c (3.20 g, 14.0 mmol, 39.3%).

[1005] 1 H NMR (400MHz, CD3OD) δ7.50-7.59(m,2H),7.27-7.35(m,2H),4.57(s,2H),1.28(s,9H).

[1006] 3. Preparation of Compound AE5

[1007] Compound 3c (1.00 g, 4.38 mmol) was dissolved in DCM (10.0 mL). The reaction mixture was cooled to 0°C, and DCI (0.57 g, 4.8 mmol) and compound 3d (2.64 g, 8.76 mmol) were added. The reaction was allowed to react at 0°C for 1 hour. TLC (petroleum ether:ethyl acetate = 3:1) indicated the disappearance of the starting material. 0.3 mL of triethylamine and basic silica gel were added to the reaction mixture, and the mixture was spin-dried. The crude product was purified by column chromatography (petroleum ether:ethyl acetate) to afford compound AE5 (1.10 g, 2.57 mmol, 58.61%).

[1008] 1 H NMR (400MHz, CDCl3) δ7.51-7.56(m,2H),7.25-7.35(m,2H),4.55-4.80(m,2H),3.76- 3.93(m,2H),3.58-3.71(m,2H),2.60-2.66(m,2H),1.30(s,9H),1.16-1.23(m,12H).

[1009] Example 8 Preparation of siRNA

[1010] The siRNA of the present invention is prepared using the solid phase phosphoramidite method well known in the art. Specific methods can be found in, for example, PCT Publication Nos. WO2016081444 and WO2019105419, and are briefly described below.

[1011] 1 Synthesis of positive sense chain (SS chain)

[1012] Using solid-phase phosphoramidite synthesis, a blank CPG solid support or a solid support connected to L96 is used as the starting cycle. Nucleoside monomers are attached one by one in the 3'-5' direction according to the order of the positive chain nucleotides. Each ligation of a nucleoside monomer involves four steps: deprotection, coupling, capping, and oxidation or thiolation. The synthesis conditions for a 5 μmol oligonucleotide are as follows:

[1013] The nucleoside monomers were provided in a 0.05 mol / L acetonitrile solution. The reaction conditions for each step were the same, i.e., the temperature was 25°C. Deprotection was performed three times using a 3% trichloroacetic acid-dichloromethane solution. The coupling reaction was performed twice using a 0.25 mol / L 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 / tetrahydrofuran / pyridine / water (70 / 20 / 10, v / v / v). Thiolysis was performed twice using 0.2 mol / L PADS in acetonitrile / 3-methylpyridine (1 / 1, v / v).

[1014] 2 Synthesis of antisense strand (AS strand)

[1015] Using solid-phase phosphoramidite synthesis, a blank CPG solid support was used as the starting cycle. Nucleoside monomers or nucleotide duplexes of the present invention were linked one by one in the 3'-5' direction according to the order of the antisense nucleotides. Each ligation involved four steps: deprotection, coupling, capping, and oxidation or thiolation. The synthesis conditions for 5 μmol of the antisense oligonucleotide were the same as those for the sense strand.

[1016] 3. Oligonucleotide Purification and Annealing

[1017] 3.1 Ammonolysis

[1018] The synthesized solid phase carrier (sense chain or antisense chain) was added to a 5 mL centrifuge tube, 3% diethylamine / ammonia water (v / v) was added, and the reaction was carried out in a constant temperature water bath at 35°C (or 55°C) for 16 hours (or 8 hours). After filtering, the solid phase carrier was washed three times with ethanol / water, each time with 1 mL. The filtrate was centrifuged and concentrated, and the crude product was purified.

[1019] 3.2 Purification

[1020] 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.

[1021] 3.3 Annealing

[1022] According to the table below, the sense strand (SS strand) and the antisense strand (AS strand) were mixed at a molar ratio (SS strand / AS strand = 1 / 1.05), heated in a water bath to 70-95°C for 3-5 minutes, cooled naturally to room temperature, and the system was lyophilized to obtain the product.

[1023] The siRNA sequences used in the present invention are as follows:

[1024] In this article, the meanings of the abbreviations are as follows:

[1025] The A, U, G, and C distribution represents the natural adenine, uracil, guanine, and cytosine ribonucleotides.

[1026] 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.

[1027] 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.

[1028] "s" or "s-" indicates that the two adjacent nucleotides and / or delivery vectors are linked by phosphorothioate.

[1029] L96 represents a GalNAc delivery vector of the following structure well known in the art, wherein The position of the siRNA is indicated by a phosphate group or a phosphorothioate group, as described in, for example, PCT Publication Nos. WO2009073809 and WO2009082607.

[1030] RPD1-RPD5 represents the following structure, where Represents the chemical bond connecting to the hydroxyl group of the 5' carbon atom of the ribose sugar of the right nucleotide:

[1031] Example 9 C57BL / 6 wild-type mouse primary hepatocyte (PMH) activity screening experiment

[1032] 1. Free uptake or transfection

[1033] Isolate C57BL / 6 wild-type mouse liver primary cells, count, and plate in 24-well plates, 900 μL / well, 8×10 4 cells / well; 96-well plate, 100 μL / well, 1×104 cells / well. Then choose free uptake or transfection.

[1034] Free uptake: Add 10 μL of diluted compound to 90 μL of Opti-MEM, mix well, add to corresponding wells, and incubate in a 37°C, 5% CO2 incubator for 24 hours. No siRNA was added to the control group.

[1035] Transfection: Add 10 μL of diluted compound to 40 μL of Opti-MEM and mix thoroughly. Add 3 μL of RNAiMAX to 47 μL of Opti-MEM and mix thoroughly. Incubate for 5 minutes, then mix with the diluted compound. Let stand at room temperature for 10 minutes, then add to the corresponding wells and incubate in a 37°C, 5% CO2 incubator for 24 hours. No siRNA was added to the control group.

[1036] 2. Fluorescence quantitative PCR

[1037] Total RNA was extracted using a high-throughput nucleic acid extraction instrument-magnetic bead method (Fanzhi Medical, FG0412; Hangzhou Aosheng, Auto-pure96), and reverse transcribed (PrimeScript TM II 1st Strand cDNA Synthesis Kit (Takara, 6210B) and then fluorescent quantitative PCR detection (TaqMan TM Fast Advanced Master Mix (ABI, 4444965)).

[1038] Table 1 Primer information

[1039] 3. Data Statistics

[1040] Calculation 2 -△△Ct The values ​​were converted into percentages to obtain the residual inhibition rate;

[1041] △△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)].

[1042] The target gene is mAPOB or mFXII, and the control gene is mGAPDH.

[1043] 4. mAPOB gene experimental results

[1044] Using the transfection method, C57BL / 6 wild-type mouse primary hepatocytes were selected. After plating the cells in 24-well plates, the starting concentration of the compound was selected to be 40 nM, and 5 concentration points (40 nM, 4 nM, 0.4 nM, 0.04 nM, 0.004 nM) were diluted 10-fold to perform a 5-point IC50 activity screening of C57BL / 6 wild-type mouse primary hepatocytes. The experimental results are shown in Table 2.

[1045] Table 2 Results of the 5-point IC50 activity screening experiment of the siRNA compound PMH targeting the mAPOB gene carrying the compound of the present invention

[1046] 5. mFXII gene experimental results

[1047] Using the free uptake method, C57BL / 6 wild-type mouse primary hepatocytes were selected, and after plating the cells in 96-well plates, the compound was selected with a starting concentration of 100 nM, and 5 concentration points (100 nM, 10 nM, 1 nM, 0.1 nM, 0.01 nM) were diluted 10-fold to perform a 5-point IC50 activity screening of C57BL / 6 wild-type mouse primary hepatocytes - free uptake. The experimental results are shown in Table 3.

[1048] Table 3 Results of the 5-point IC50 activity screening experiment of the siRNA compound PMH targeting the mFXII gene carrying the compound of the present invention

[1049] Example 10 Verification of compound efficacy in C57BL / 6 mouse model

[1050] C57BL / 6 mice (male, 18-21 g, 6-8 weeks, Sibeifu (Suzhou) Biotechnology Co., Ltd.) were randomly divided into groups of 9 animals per group (mice were euthanized on the 14th and 28th days (N=3 / group) for liver removal). The dosage of each animal was calculated based on body weight, and a single dose was administered subcutaneously. The siRNA conjugate was provided to the CRO company as a 10 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 diluted to the required solution concentration and volume. The administration volume of physiological saline and siRNA conjugate was 5 mL / kg.

[1051] Blood was collected before administration (designated as day -2) for testing. Groups were divided based on LDL levels on day 2, with the remaining samples retained for target protein detection. Blood was collected from the orbital venous plexus of mice on days 7, 14, 21, 28, and 42 after administration (designated as day 0) (starved for 5 hours before each blood draw). Serum LDL concentrations (Meikang brand, low-density lipoprotein cholesterol detection reagent) were measured by Suzhou Anling at each time point using a direct method (Neusoft fully automatic biochemical analyzer, NT-1000). The remaining samples were retained for ELISA detection of the target protein (Mouse ApoB ELISA Kit, Abcam, ab230932). On days 14, 28, and 42, 10 mg of liver (n = 3 / group / time point) was collected, placed in RNAlater solution, frozen at -80°C, and shipped on dry ice for extraction and detection of liver mRNA expression (see Table 4 for detection primers, which will be updated at subsequent time points). The experimental results are shown in Table 5.

[1052] Table 4 Primer information

[1053] Table 5 Pharmacological efficacy of siRNA compounds in C57BL / 6 mouse model

[1054] Example 11 C57BL / 6 wild-type mouse primary hepatocyte (PMH) activity screening experiment

[1055] According to the method of Example 9, more compounds of the present invention were tested.

[1056] Table 6. Results of the 5-point IC50 activity screening experiment of the siRNA compound PMH targeting the mAPOB gene carrying the compound of the present invention

[1057] Table 7. Results of the 5-point IC50 activity screening experiment of the siRNA compound PMH targeting the mFXII gene carrying the compound of the present invention

Claims

1. An oligonucleotide comprising a compound of formula (II), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof: in, X1 is selected from OR a or NR b R c ; R a Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated; R b and R c Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration; X2 is the remainder of the oligonucleotide, which is linked to P through the hydroxyl or sulfhydryl group on the 2', 3' or 5' carbon atom of the ribose of the first nucleotide at the 5' end; X3 is independently selected from O or S; Each R T1 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated; Each R T2 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated; m is 0, 1, 2, 3, 4 or 5; X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -; R X1 Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated; L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by 1 or 2 R#, R# is selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated; Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom; Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2, 3, 4 or 5 R*; R*selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, which is optionally deuterated, up to fully deuterated.

2. The oligonucleotide of claim 1, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein X1 is selected from OR a or NR b R c ; R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; R b and R c Independently selected from H, C 1-4 Alkyl, C 1-4 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl substitution until complete deuteration; X2 is the remainder of the oligonucleotide, which is linked to P through the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose of the first nucleotide at the 5' end; X3 is independently selected from O or S; Each R T1 Independently selected from H, D, C 1-4 Alkyl, C 1-4 a haloalkyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated; Each R T2 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; m is 0, 1, 2, or 3; X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -; R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; L is -Ar-(CH2)-O-, wherein CH2 may be optionally substituted by 1 or 2 R#, R# being selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom; Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*; R*selected from C 1-4 Alkyl or C 1-4 Haloalkyl, which is optionally deuterated, up to fully deuterated.

3. The oligonucleotide of claim 1 or 2, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: X1 is selected from OR a or NR b R c ; R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; R b and R c Independently selected from H, C 1-4 Alkyl, C 1-4 haloalkyl, the R b and R c Optionally substituted with D, phenyl, up to full deuteration; X2 is the remainder of the oligonucleotide, which is linked to P through the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose of the first nucleotide at the 5' end; X3 is independently selected from O or S; Selected from where R T is selected from H, D, CH3 or a chain comprising GalNAc, which is optionally deuterated, up to fully deuterated; X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -; R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; L is selected from 4. The oligonucleotide according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein X1 is selected from OH or X2 is the remainder of the oligonucleotide, which is linked to P through the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose of the first nucleotide at the 5' end; X3 is independently selected from O or S; Selected from X is selected from a chemical bond, -O-, -NHC(O)O-, -OC(O)NH-, -N(CH3)C(O)O- or -C(O)O-; L is selected from 5. The oligonucleotide according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the compound of formula (II) is selected from: in, X1 is selected from OH or X2 is the remainder of the oligonucleotide, which is linked to P via the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the first nucleotide at the 5' end; X3 is independently selected from O or S; Preferably, the compound of formula (I) is selected from: X2 is the remainder of the oligonucleotide, which is linked to P via the hydroxyl group on the 5' carbon atom of the ribose sugar of the first nucleotide at the 5' end.

6. An oligonucleotide comprising one, two or more compounds of formula (Ia), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof: in, Selected from OR a or NR b R c , or represents a chemical bond connecting to the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide; X1 is a chemical bond connecting to the hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose of another adjacent nucleotide; R a Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated; R b and R c Independently selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, the R b and R c Can be optionally D, C 6-10 Aryl or 5-10 membered heteroaryl substitution, up to full deuteration; X3 is independently selected from O or S; T is Each R T1 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated; Each R T2 Independently selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated; m is 0, 1, 2, 3, 4 or 5; X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -; R X1 Selected from H, halogen, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated; L is -Ar-(CH2) 1-6 -O-, wherein each CH2 may be optionally substituted by R#, R# is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated; Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom; Ar is selected from C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-14 membered heteroaryl, the C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2, 3, 4 or 5 R*; R* is selected from H, D, halogen, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl, which is optionally deuterated, up to fully deuterated.

7. The oligonucleotide according to claim 6, wherein Selected from OR a or NR b R c , or represents a chemical bond connecting to the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide; X1 is a chemical bond connecting to the hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose of another adjacent nucleotide; R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; R b and R c Independently selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, and the R b and R c Can be optionally D, C 6-10 aryl substitution, which is optionally deuterated, up to fully deuterated; X2 and X3 are independently selected from O or S; T is selected from Each R T1 Independently selected from H, D, C 1-4 Alkyl, C 1-4 A haloalkyl or GalNAc-containing chain, which is optionally deuterated, to complete deuteration; Each R T2 Independently selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; m is 0, 1, 2, or 3; X is selected from a chemical bond, -O-, -S-, -OC(O)NR X1 -、-NR X1 C(O)O-, -C(O)O-, -OC(O)-, -NR X1 C(O)- or -C(O)NR X1 -; R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; L is -Ar-(CH2)-O-, wherein CH2 may be optionally substituted by 1 or 2 R#, R# being selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom; Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*; R*selected from C 1-4 Alkyl or C 1-4 Haloalkyl, which is optionally deuterated, up to fully deuterated.

8. The oligonucleotide according to claim 6 or 7, wherein Selected from OR a or NR b R c , or represents a chemical bond connecting to the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide; X1 is a chemical bond connecting to the hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose of another adjacent nucleotide; R a Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; R b and R c Independently selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, the R b and R c Optionally substituted with D, phenyl, up to full deuteration; X3 is independently selected from O or S; T is selected from where R T is selected from H, D, CH3 or a chain comprising GalNAc, which is optionally deuterated, up to fully deuterated; X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -; R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; L is selected from 9. The oligonucleotide according to any one of claims 6 to 8, wherein It represents a chemical bond to the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide; X1 is a chemical bond connecting to the hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose of another adjacent nucleotide; X3 is selected from O or S; T is selected from X is selected from a chemical bond, -O-, -NHC(O)O-, -OC(O)NH-, -N(CH3)C(O)O- or -C(O)O-; L is selected from 10. The oligonucleotide according to any one of claims 6 to 9, wherein The compound of formula (Ia) is selected from the following structures: in, It represents a chemical bond to the hydroxyl or sulfhydryl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide; X1 is a chemical bond connecting to the hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose of another adjacent nucleotide; X3 is selected from O or S; Preferably, the compound of formula (Ia) is selected from: It represents the chemical bond connecting to the hydroxyl group on the 5' carbon atom of the ribose sugar of the adjacent nucleotide; X1 is a chemical bond connecting to the hydroxyl or sulfhydryl group on the 2' or 3' carbon atom of the ribose of another adjacent nucleotide; X3 is selected from O or S.

11. The oligonucleotide of any one of claims 6 to 10, having 14 to 30 nucleotides.

12. The oligonucleotide according to any one of claims 6 to 11, comprising one or more compounds of formula (Ia), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, within the oligonucleotide.

13. A double-stranded RNA having a sense strand and an antisense strand, each strand having 14 to 30 nucleotides, the antisense strand comprising a sequence that is sufficiently complementary to the sense strand and target mRNA, wherein the sense strand and / or antisense strand comprises one or more compounds of formula (II) according to any one of claims 1 to 5 or compounds of formula (Ia) according to any one of claims 6 to 10, or pharmaceutically acceptable salts, tautomers, or stereoisomers thereof.

14. The double-stranded RNA of claim 13, wherein the sense strand comprises one or more compounds of formula (Ia) according to any one of claims 6 to 10, or pharmaceutically acceptable salts, tautomers or stereoisomers thereof, within the oligonucleotide.

15. The double-stranded RNA according to claim 13, wherein the antisense strand comprises a compound of formula (II) according to any one of claims 1 to 5, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof at the 5' end.

16. The double-stranded RNA of claim 13, wherein the antisense strand comprises one or more compounds of formula (Ia) according to any one of claims 6 to 10, or pharmaceutically acceptable salts, tautomers or stereoisomers thereof, within the oligonucleotide.

17. The double-stranded RNA according to any one of claims 13 to 16, which is selected from small interfering RNA (siRNA) and short hairpin RNA (shRNA).

18. A cell comprising the double-stranded RNA according to any one of claims 14 to 17.

19. A pharmaceutical composition comprising the double-stranded RNA according to any one of claims 14 to 17, or the cell according to claim 18, and optionally a pharmaceutically acceptable carrier or excipient.

20. A kit comprising the double-stranded RNA according to any one of claims 14 to 17, the cell according to claim 18, or the pharmaceutical composition according to claim 19.

21. A compound of formula (IIb), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof: in, Each R T1 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 an alkynyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated; Each R T2 Independently selected from H, D, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated; m is 0, 1, 2, 3, 4 or 5; X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -; R X1 Selected from H, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated; L is -Ar-(CH2)-O-, wherein CH2 may be optionally substituted by 1 or 2 R#, R# being selected from H, D, C 1-6 Alkyl or C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated; Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*; R*selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, which is optionally deuterated, up to fully deuterated; PG is selected from protecting groups.

22. The compound of claim 21, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein Each R T1 Independently selected from H, D, C 1-6 Alkyl, C 1-6 a haloalkyl group or a GalNAc-containing chain, which is optionally deuterated, up to fully deuterated; Each R T2 Independently selected from H, D, C 1-6 Alkyl, C 1-6 haloalkyl, which is optionally deuterated, up to fully deuterated; m is 0, 1, 2, or 3; X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)NR X1 -、-NR X1 C(O)O-、-NR X1 C(O)- or -C(O)NR X1 -; R X1 Selected from H, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; L is -Ar-(CH2)-O-, wherein CH2 may be optionally substituted by 1 or 2 R#, R# being selected from H, D, C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; Ar is selected from C 6-10 Aryl or 5-10 membered heteroaryl, the C 6-10 Aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2 or 3 R*; R*selected from C 1-4 Alkyl or C 1-4 haloalkyl, which is optionally deuterated, up to fully deuterated; PG is selected from protecting groups.

23. The compound of claim 21 or 22, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein PG is selected from 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), 4,4'-dimethoxytrityl, -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH, preferably -P(OCH2CH2CN)(N(iPr)2) or -C(O)CH2CH2C(O)OH.

24. The compound of any one of claims 21 to 23, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein The compound of formula (IIb) is selected from: in, PG is -P(OCH2CH2CN)(N(iPr)2) or H.

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