Three-cluster galactose compound, conjugate as well as preparation method and application of three-cluster galactose compound

Abstract

The invention relates to a three-cluster galactose compound, a conjugate as well as a preparation method and application of the three-cluster galactose compound. Specifically, the invention relates to compounds of formula (I) and conjugates of the compounds and drug molecules. The invention also relates to a preparation method of the compound, and application of the compound and the conjugate in preparation of drugs for treating or preventing diseases related to expression or overexpression of genes in hepatocytes. # imgabs0 #

Description

Field of the Invention

[0001] The present invention relates to the field of biotechnology, and in particular to a gel preparation of a tri-cluster galactose compound and a conjugate. The present invention also relates to a method for preparing the compound, and the use of the compound and the conjugate in preparing a drug for treating or preventing diseases related to the expression or overexpression of genes in hepatocytes. Background Art

[0002] Asialoglycoprotein receptor (ASGPR) is an abundant heterologous oligomeric endocytic receptor, mainly present on the cell membrane surface of liver parenchymal cells facing the sinusoidal space, with specific recognition of sugars. Since the terminal sialic acid of various glycoproteins is removed by enzymatic hydrolysis or acidolysis, the exposed secondary end is a galactose residue, so the sugar binding specificity of ASGPR is actually galactose, so it is also called galactose specific receptor. ASGPR is mainly distributed in liver parenchymal cells, and its content in other cells is low, so it has become the best receptor for liver-directed transport.

[0003] Glycoproteins with non-reducing galactose (Gal) or N-acetylgalactosamine (GalNAc) residues at the end can be recognized by ASGPR, and the affinity of GalNAc binding to ASGPR is about 50 times higher than that of Gal (Lobst ST et al, J Biol Chem, 1996, 271(12): 6686-6693). In vitro experiments have shown that clustered sugar residues can occupy the binding sites of the receptor at the same time, making their affinity much higher than that of non-clustered sugar residues.

[0004] ASGPR receptor-mediated liver-targeted oligonucleotides are a new breakthrough in the field of innovative nucleic acid drug research. Among them, GalNAc (N-acetylgalactosamine) coupling modification is the most commonly used small nucleic acid drug delivery system. At present, most of the conjugate structures of GalNAc are tri-contact GalNAc structures, which specifically refers to the covalent conjugation of GalNAc to the 3' or 5' end of the sense chain of siRNA of different sequences in a trivalent state to form a polysaccharide-siRNA monoconjugate, thereby achieving specific delivery to hepatocytes and allowing the drug to enter the cell to exert its function through endocytosis.

[0005] In 2012, Alnylam Pharmaceuticals in the United States covalently linked the triantennary GalNAc structure studied by previous researchers with small interfering RNA (siRNA), achieving liver-targeted delivery of siRNA in vivo. Using this technology, researchers have developed drugs for diseases such as amyloidosis, hemophilia, hypercholesterolemia, liver porphyrin, and hepatitis B. In 2014, ISIS Pharmaceuticals in the United States covalently linked triantennary GalNAc with antisense nucleic acid to achieve liver-targeted drug delivery in animals. After the connection, the activity of the antisense nucleic acid increased by 10 times (Prakash TP et al, Nucleic Acids Res. 42, 8796-807).

[0006] However, ASGPR receptor-mediated liver-targeted oligonucleotide technology still needs further development and improvement, and people are still actively studying other structures that are different from the traditional three-touch GalNAc structure. Summary of the invention

[0007] The present invention aims to solve the above problems and provide a three-cluster galactose compound that is different from the traditional three-touch GalNAc structure, can maintain good liver targeting affinity, has good liver targeting, has better hydrophilicity and low toxicity, and is easy to synthesize.

[0008] In order to develop a new structure of three-cluster galactose compounds, the inventors tried to use a variety of amino acids to synthesize compounds with different branched structures, such as glutamic acid, aspartic acid or lysine, but these structures did not significantly improve the ASGPR receptor affinity. After continuous attempts and research, the inventors found that the structure formed by the condensation of L-serine and serinol as a branched structure, the resulting three-cluster galactose compound has better hydrophilicity and low toxicity, and has a strong application value in clinical practice.

[0009]

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

[0011]

[0012] wherein each group is as defined below.

[0013] In another aspect, the present invention provides a conjugate having a structure shown in formula (III):

[0014]

[0015] wherein each group is as defined below.

[0016] In another aspect, the present invention provides a pharmaceutical composition comprising the conjugate of the present invention, and optionally a pharmaceutically acceptable carrier, excipient, adjuvant or vehicle, and optionally other therapeutic agents.

[0017] In another aspect, the present invention provides use of the conjugate of the present invention in the preparation of a medicament for treating or preventing a disease associated with the expression or overexpression of a gene in hepatocytes.

[0018] In another aspect, the present invention provides a method for treating or preventing a disease associated with the expression or overexpression of a gene in hepatocytes in a subject, comprising administering the conjugate of the present invention or the pharmaceutical composition of the present invention to the subject.

[0019] In another aspect, the present invention provides a conjugate of the present invention or a pharmaceutical composition of the present invention for use in treating or preventing a disease associated with the expression or overexpression of a gene in hepatocytes.

[0020] In a specific embodiment, the gene is selected from the HBV genome, the HCV genome, PCSK9, xanthine oxidase, URAT1, APOB, liver fibrosis-related genes (AP3S2, AQP2, AZINI, DEGSI, STXBP5L, TLR4, TRPM5), non-alcoholic fatty liver disease-related genes (PNPLA3, FDFTl), primary biliary cirrhosis-related genes (HLA-DQB1, IL-12, IL-12RB2), or a combination thereof.

[0021] In another specific embodiment, the present invention is used to treat or prevent the following diseases: hereditary angioedema, familial tyrosinemia type I, Alagille syndrome, alpha-1-antitrypsin deficiency, defects in bile acid synthesis and metabolism, biliary atresia, cystic fibrosis liver disease, idiopathic neonatal hepatitis, mitochondrial liver disease, progressive familial intrahepatic cholestasis, primary sclerosing cholangitis, transthyretin amyloidosis, hemophilia, homozygous familial hypercholesterolemia, hyperlipidemia, hepatitis B (HBV), hepatitis C (HCV), steatohepatitis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), hyperglycemia or diseases involving abnormally increased hepatic glucose production similar to type III diabetes, hepatitis, and hepatic porphyrins.

[0022] definition

[0023] Chemical Definition

[0024] Definitions of specific functional groups and chemical terms are described in more detail below.

[0025] 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.

[0026] “C 1-10 "Alkyl" refers to a straight or branched chain saturated hydrocarbon group having 1 to 10 carbon atoms. In some embodiments, C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C 1-3 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 by one or more substituents, for example, by 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).

[0027] “C 2-6 "Alkenyl" refers to a straight or branched 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 “C2-6 "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). The alkenyl group may be optionally substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0028] “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. 2-6 Examples of alkynyl groups include, but are not limited to, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), pentynyl (C5), hexynyl (C6), and the like. The term “C 2-6 "Alkynyl" also includes heteroalkynyl groups, in which one or more (e.g., 1, 2, 3 or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). Alkynyl groups may be optionally substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0029] “C 1-25 "Alkylene" refers to the removal of C 1-25 The other hydrogen of C alkyl forms a divalent group, and may be substituted or unsubstituted. 5-25 Alkylene, C 1-20 Alkylene, C 3-20 Alkylene, C 5-20 Alkylene, C 8-20 Alkylene, C 10-17 Alkylene, C 1-17 Alkylene, C 3-17 Alkylene, C 1-10 Alkylene, C 3-10 Alkylene, C 1-6 Alkylene, C 3-6 Alkylene, C 4-6 Alkylene, C 1-4 Alkylene, C 2-4 Alkylene and C 1-3Alkylene is preferred. In some embodiments, C6 alkylene and C4 alkylene are preferred. Unsubstituted alkylene includes, but is not limited to, methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), butylene (-CH2CH2CH2CH2-), pentylene (-CH2CH2CH2CH2CH2-), hexylene (-CH2CH2CH2CH2CH2CH2-), and the like. Exemplary substituted alkylenes, for example, alkylenes substituted with one or more alkyl (methyl) groups, include, but are not limited to, substituted methylene (-CH(CH3)-, -C(CH3)2-), substituted ethylene (-CH(CH3)CH2-, -CH2CH(CH3)-, -C(CH3)2CH2-, -CH2C(CH3) 2- ), substituted propylene (-CH(CH3)CH2CH2-, -CH2CH(CH3)CH2-, -CH2CH2CH(CH3)-, -C(CH3)2CH2CH2-, -CH2C(CH3)2CH2-, -CH2CH2C(CH3)2-), and the like. In some embodiments, straight chain alkylene is preferred.

[0030] “C 0-6 "Alkylene" refers to a chemical bond and the above-mentioned "C 1-6 Alkylene", "C 0-4 "Alkylene" refers to a chemical bond and the above-mentioned "C 1-4 "Alkylene".

[0031] "Halo" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

[0032] Therefore, “C 1-10 "Haloalkyl" refers to the above-mentioned "C 1-10 "alkyl", which is substituted by one or more halogen groups. In some embodiments, C 1-6 Haloalkyl, C 1-5 Haloalkyl, C 1-4 Haloalkyl and C 1-3 Haloalkyl is particularly preferred, more preferably C 1-2 Haloalkyl. Exemplary haloalkyls 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 may be substituted at any available point of attachment, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0033] “C 3-10"Cycloalkyl" refers to a non-aromatic cyclic hydrocarbon group having 3 to 10 ring carbon atoms and zero heteroatoms, optionally containing 1, 2 or 3 double or triple bonds. In some embodiments, C 5-10 Cycloalkyl, C 3-7 Cycloalkyl and C 3-6 Cycloalkyl is particularly preferred, more preferably C 5-7 Cycloalkyl and C 5-6 Cycloalkyl. Cycloalkyl also includes a ring system in which the above-mentioned cycloalkyl ring is fused with 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. Cycloalkyl also includes the above-mentioned cycloalkyl ring, wherein the substituents on any non-adjacent carbon atoms are connected to form a bridge ring, together forming a polycycloalkane sharing two or more carbon atoms. Cycloalkyl also includes the above-mentioned cycloalkyl ring, wherein the substituents on the same carbon atom are connected to form a ring, together forming a polycycloalkane sharing one carbon atom. 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. A 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.

[0034] "3-10 membered heterocyclyl" refers to a saturated or unsaturated radical of a 3- to 10-membered non-aromatic ring system having ring carbon atoms and 1 to 5 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon, and optionally containing 1, 2 or 3 double or triple bonds. In heterocyclyls containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom as long as valence permits. In some embodiments, a 5-10 membered heterocyclyl is preferred, which is a 5-10 membered non-aromatic ring system having ring carbon atoms and 1 to 5 ring heteroatoms; in some embodiments, a 3-7 membered heterocyclyl is preferred, which is a 3-7 membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms; a 5-7 membered heterocyclyl is preferred, which is a 5-7 membered non-aromatic ring system having ring carbon atoms and 1 to 3 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 4-6 membered heterocyclyl is preferred, which is a 4-6 membered non-aromatic ring system having ring carbon atoms and 1 to 3 ring heteroatoms; and a 5-6 membered heterocyclyl is more preferred, which is a 5-6 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 with one or more cycloalkyl groups, wherein the point of attachment is on the heterocyclyl ring, or a ring system in which the above-mentioned heterocyclyl ring is fused with 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. Heterocyclyl also includes the above-mentioned heterocyclyl ring, wherein the substituents on any non-adjacent carbon or nitrogen atoms are connected to form a bridge ring, together forming a polycyclic heteroalkane sharing two or more carbon or nitrogen atoms. Heterocyclyl also includes the above-mentioned heterocyclyl ring, wherein the substituents on the same carbon atom are connected to form a ring, together forming a polycyclic heteroalkane sharing one carbon atom. Exemplary 3-membered heterocyclyls containing one heteroatom include, but are not limited to: aziridine, oxadiazine, thiorenyl. Exemplary 4-membered heterocyclyls containing one heteroatom include, but are not limited to: azetidinyl, oxadiazine and thiazine. Exemplary 5-membered heterocyclic groups containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to, pyrazolidinyl, dioxolanyl, oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-membered heterocyclic groups containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing one heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl.Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithiohexane, dioxanyl. Exemplary 6-membered heterocyclic groups containing three heteroatoms include, but are not limited to, hexahydrotriazine (triazinanyl). Exemplary 7-membered heterocyclic groups containing one heteroatom include, but are not limited to, azepanyl, oxepane and thiepanyl. Exemplary 5-membered heterocyclic groups fused to a C6 aryl ring (also referred to herein as 5,6-bicyclic heterocyclic groups) include, but are not limited to, dihydroindole, isoindole, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinone, and the like. Exemplary 6-membered heterocyclic groups fused to a C6 aryl ring (also referred to herein as 6,6-bicyclic heterocyclic groups) include, but are not limited to, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like. Heterocyclyl also includes the above heterocyclyl and a cycloalkyl, heterocyclyl, aryl or heteroaryl sharing one or two atoms to form a bridged ring or a spirocycle, and the shared atom can be a carbon or nitrogen atom as long as the valence permits. Heterocyclyl also includes the above heterocyclyl and heterocyclyl groups can be optionally substituted by one or more substituents, for example, substituted by 1 to 5 substituents, 1 to 3 substituents or 1 substituent.

[0035] “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, aryl has six ring carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, aryl has ten ring carbon atoms ("C 10 Aryl also includes ring systems in which the above aryl ring 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, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0036] "5-10 membered heteroaryl" refers to a group of a 5-10 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 the valence permits. Heteroaryl bicyclic ring systems may include one or more heteroatoms in one or both rings. Heteroaryl also includes a ring system in which the above-mentioned heteroaryl ring is fused to one or more cycloalkyl or heterocyclic 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 other embodiments, 5-6 membered heteroaryl groups are particularly preferred, which are 5-6 membered monocyclic or bicyclic 4n+2 aromatic ring systems having ring carbon atoms and 1-4 ring heteroatoms. Exemplary 5-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyrrolyl, furanyl, and thienyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl (e.g., 1,2,4-oxadiazolyl), and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridyl or pyridone. Exemplary 6-membered heteroaryl groups 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, oxacycloheptatrienyl, and thiacycloheptatrienyl. Exemplary 5,6-bicyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothienyl, isobenzothienyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indazinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, but are not limited to, naphthyridinyl, pteridinyl, quinolyl, isoquinolyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. The heteroaryl group may be optionally substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0037] “C 6-10 "Arylene" refers to the removal of C 6-10 The other hydrogen of the aryl group forms a divalent group, and may be substituted or unsubstituted. In some embodiments, phenylene is preferred, for example

[0038] “C5-10 "Heteroarylene" refers to a C 5-10 The divalent group is formed by the other hydrogen of the heteroaryl group and may be substituted or unsubstituted. 5-9 Heteroarylene, C 5-6 Heteroarylene, C6 heteroarylene and C5 heteroarylene are particularly preferred, for example For example

[0039] The divalent groups formed by removing another hydrogen from the above-defined alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups are collectively referred to as "subunits". The ring-forming groups such as cycloalkyl, heterocyclyl, aryl and heteroaryl groups are collectively referred to as "cyclyls".

[0040] Alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups and the like as defined herein are optionally substituted groups.

[0041] 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 )ORaa 、-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、-BRaa (OR cc ), alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R dd Group substitution;

[0042] 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;

[0043] 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 substituted by 0, 1, 2, 3, 4 or 5 R dd Group substitution;

[0044] 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)(NRcc )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 substituted by 0, 1, 2, 3, 4 or 5 R dd Group substitution;

[0045] 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 substituted by 0, 1, 2, 3, 4 or 5 R dd Group substitution;

[0046] R dd Each of 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(Rff )2.-NR ff C(=NR ff )N(R ff )2.-NR ff S02R ee 、-SO2N(R ff 2. -SO2R ee 、-SO2OR ee 、-OSO2R ee 、-S(=O)R ee 、-Si(R ee )3.-OSi(R ee )3. -C(=S)N(R ff )2, -C(=O)SR ee 、-C(=S)SR ee 、-SC(=S)SR ee 、-P(=O)2R ee ,-P(=O)(R ee )2.-OP(=O)(R ee )2.-OP(=O)(OR ee )2, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 R gg group substitution, or two geminal R dd Substituents may combine to form =O or =S;

[0047] 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 substituted by 0, 1, 2, 3, 4 or 5 R gg Group substitution;

[0048] 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 substituted by 0, 1, 2, 3, 4 or 5 R gg Group substitution;

[0049] R gg Each of is independently: halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OC 1-6 Alkyl, -ON(C 1-6Alkyl)2, -N(C 1-6 Alkyl)2, -N(C 1-6 Alkyl)3 + X - 、-NH(C 1-6 Alkyl)2 + X - 、-NH2(C 1-6 alkyl) + X - 、-NH3 + X - 、-N(OC 1-6 Alkyl)(C 1-6 Alkyl), -N(OH)(C 1-6 Alkyl), -NH(OH), -SH, -SC 1-6 Alkyl, -SS(C 1-6 alkyl), -C(=O)(C 1-6 alkyl), -CO2H, -CO2(C 1-6 alkyl), -OC(=O)(C 1-6 Alkyl), -OCO2(C 1-6 alkyl), -C(=O)NH2, -C(=O)N(C 1-6 alkyl)2, -OC(=O)NH(C 1-6 alkyl), -NHC(=O)(C 1-6 Alkyl), -N(C 1-6 alkyl)C(=O)(C 1-6 Alkyl), -NHCO2(C 1-6 alkyl), -NHC(=O)N(C 1-6 Alkyl)2, -NHC(=O)NH(C 1-6 alkyl), -NHC(=O)NH2, -C(=NH)O(C 1-6 alkyl), -OC(=NH)(C 1-6 alkyl), -OC(=NH)OC 1-6 Alkyl, -C(=NH)N(C 1-6 Alkyl)2, -C(=NH)NH(C 1-6 alkyl), -C(=NH)NH2, -OC(=NH)N(C 1-6 Alkyl)2, -OC(NH)NH(C 1-6 alkyl), -OC(NH)NH2, -NHC(NH)N(C 1-6 Alkyl)2, -NHC(=NH)NH2, -NHSO2(C 1-6 Alkyl), -SO2N(C 1-6 Alkyl)2, -SO2NH(C 1-6Alkyl), -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 1-6 Alkyl)2, -OP(=O)(OC 1-6 Alkyl)2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, C6-C 10 Aryl, C3-C7 heterocyclic, C5-C 10 heteroaryl; or two geminal R gg Substituents may combine to form =O or =S; wherein X - For the counter ion.

[0050] 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)2Raa ,-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 substituted by 0, 1, 2, 3, 4 or 5 R dd substituted with a group, and wherein R aa , R bb , R cc and R dd As mentioned above.

[0051] Other definitions

[0052] The term "oligonucleotide chain" refers to an oligomeric compound containing multiple or all chemically modified or unmodified nucleotides, which has a length of less than about 100 nucleotides (e.g., 1-20 nucleotides or 1-50 nucleotides). In certain embodiments, a non-nucleic acid conjugated group may be included in the oligonucleotide chain. In certain embodiments, the oligonucleotide comprises ribonucleic acid (RNA), deoxyribonucleic acid (DNA), or oligopeptide nucleotides (PNA). In certain embodiments, the oligonucleotide chain is double-stranded or single-stranded. In certain embodiments, the oligonucleotide chain is siRNA, nucleic acid aptamer, antisense nucleic acid, sgRNA, tractRNA or crRNA.

[0053] The term "conjugate group" means an atom or group of atoms that is bound to an oligonucleotide chain. In some cases, the conjugate groups alter one or more properties of the oligonucleotide to which they are attached, including but not limited to pharmacodynamics, pharmacokinetics, binding, absorption, cellular distribution, cellular uptake, charge and / or clearance properties.

[0054] The term "conjugate" means a coupled molecule of a compound of the present invention and an oligonucleotide chain.

[0055] The term "receptor" refers to a biological macromolecule composed of glycoproteins or lipoproteins, which exists in the cell membrane, cytoplasm or nucleus. Different receptors have specific structures and configurations.

[0056] The term "ligand" refers to a substance or compound that has the ability to recognize and bind to a receptor. In certain embodiments, the ligand is a ligand that binds to an asialoglycoprotein receptor (ASGPR). In certain embodiments, the ligand is a carbohydrate, such as a monosaccharide and a polysaccharide, including but not limited to galactose, N-acetylgalactosamine, mannose, glucose, glucosamine, and fucose, including but not limited to D-mannopyranose, L-mannopyranose, D-arabinose, D-xylofuranose, L-xylofuranose, D-glucose, L-glucose, D-galactose, L-galactose, α-D-mannofuranose, β-D-mannofuranose , β-D-mannopyranose, β-D-mannopyranose, α-D-glucose pyranose, β-D-glucose pyranose, α-D-glucofuranose, β-D-glucose furanose, α-D-fructofuranose, α-D-fructopyranose, α-D-galactopyranose, β-D-galactopyranose, α-D-galactofuranose, β-D-galactofuranose, glucosamine, sialic acid, galactosamine, N-acetylgalactosamine, N-trifluoroacetylgalactosamine, N- Propionylgalactosamine, N-butyrylgalactosamine, N-isobutyrylgalactosamine, 2-amino-3-O-[(R)-1-carboxyethyl]-2-deoxy-β-D-glucopyranose, 2-deoxy-2-methylamino-L-glucopyranose, 4,6-dideoxy-4-formamido-2,3-di-O-methyl-D-mannopyranose, 2-deoxy-2-sulfoamino-D-glucopyranose, N-glycolyl-α-neuraminic acid, 5-thio -β-D-glucopyranose, 2,3,4-tri-O-acetyl-1-thio-6-O-trityl-α-D-glucopyranoside methyl ester, 4-thio-β-D-galactopyranose, 3,4,6,7-tetra-O-acetyl-2-deoxy-1,5-dithio-α-D-glucopyranose heptanoside ethyl ester, 2,5-anhydro-D-allose nitrile, ribose, D-ribose, D-4-thioribose, L-ribose and L-4-thioribose.

[0057] As used herein, the term "polysaccharide" refers to a polymer formed by multiple monosaccharide groups linked by glycosidic bonds. In the present invention, polysaccharides include oligosaccharides and oligosaccharides. Generally, "oligosaccharides" refer to polymers formed by 2-10 monosaccharide groups linked by glycosidic bonds, and "oligosaccharides" refer to polymers formed by less than 20 monosaccharide groups linked by glycosidic bonds.

[0058] "Protecting group" or "protecting group" refers to any atom or group of atoms added to a molecule to prevent an existing group in the molecule from undergoing an undesired chemical reaction. A "protecting group" may be an unstable chemical moiety known in the art that is used to protect reactive groups, such as hydroxyl, amino, and thiol groups, from undesired or inappropriate reactions during chemical synthesis. Protecting groups are often 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.

[0059] 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 (Vin), methylthiomethyl ether (Mn ... 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).

[0060] "Hydroxy protecting group" refers to a group that can protect the hydroxyl group from chemical reactions and can be removed under specific conditions to restore the hydroxyl group. It mainly includes silane type protecting groups, acyl type protecting groups or ether type protecting groups, preferably the following:

[0061] Trimethylsilyl (TMS), triethylsilyl (TES), dimethylisopropylsilyl (DMIPS), diethylisopropylsilyl (DEIPS), tert-butyldimethylsilyl ether (TBDMS), tert-butyldiphenylsilyl (TBDPS), triisopropylsilyl ether (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, dimethoxytrityl (DMT), monomethoxytrityl (MMT), 9-phenylxanthen-9-yl (Pixyl), 9-(p-methoxyphenyl)xanthen-9-yl (Mox), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr), 4,4',4″-trimethoxytrityl (TMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzyloxymethyl (PMBM).

[0062] In some embodiments, the hydroxyl protecting group is preferably the following:

[0063] dimethoxytrityl (DMT), monomethoxytrityl (MMT), 9-phenylxanthen-9-yl (Pixyl), 9-(p-methoxyphenyl)xanthen-9-yl (Mox), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr) or 4,4',4"-trimethoxytrityl (TMTr), tert-butyldimethylsilyl ether (TBMDS), tert-butyldiphenylsilyl (TBDPS) and triisopropylsilyl ether (TIPS), preferably selected from dimethoxytrityl (DMT), monomethoxytrityl (MMT), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr) or 4,4',4"-trimethoxytrityl (TMTr), preferably 4,4'-dimethoxytrityl (DMTr).

[0064] In some embodiments, the hydroxyl protecting group is preferably the following:

[0065] trimethylsilyl (TMS), triethylsilyl (TES), dimethylisopropylsilyl (DMIPS), diethylisopropylsilyl (DEIPS), tert-butyldimethylsilyl ether (TBDMS), tert-butyldiphenylsilyl (TBDPS) and triisopropylsilyl ether (TIPS), preferably tert-butyldimethylsilyl ether (TBDMS).

[0066] "Carboxyl protecting group" refers to a group that can protect the carboxyl group from chemical reactions and can be removed under specific conditions to restore the carboxyl group. It mainly includes protecting groups that form esters with the carboxyl group. The carboxyl protecting group is preferably as follows:

[0067] C 1-6 Alkyl, benzyl, allyl, Preferably, the C 1-6 The alkyl group may be unsubstituted or substituted, including but not limited to: methyl, substituted methyl, β-substituted ethyl, tert-butyl, and the like.

[0068] "Amino protecting group" refers to a group that can prevent the amino group from chemical reaction and can be removed under specific conditions to restore the amino group. It includes but is not limited to: benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), 9-fluorenylmethoxycarbonyl (Fmoc), allyloxycarbonyl (Alloc), 2-(trimethylsilyl)ethoxycarbonyl (Teoc), methoxycarbonyl and ethoxycarbonyl, etc. In some embodiments, tert-butyloxycarbonyl (Boc) is preferred.

[0069] "Condensation activator" refers to a substance that increases the rate of the condensation reaction.

[0070] As used herein, the term "about" should be understood by those skilled in the art and will vary to a certain extent depending on the context in which it is used. If the meaning is not clear to one skilled in the art based on the context in which the term is used, then "about" means that the deviation does not exceed plus or minus 10% of the specified value or range.

[0071] The term "treat" as used herein relates to reversing, alleviating, inhibiting the progress of, or preventing the disorder or condition to which the term applies, or one or more symptoms of such a disorder or condition. The noun "treat" as used herein relates to the action of the verb treat, the latter being as just defined.

[0072] The term "pharmaceutically acceptable salt" as used herein refers to those carboxylates, amino acid addition salts of the compounds of the present invention which 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.

[0073] Pharmaceutically acceptable base addition salts are formed with metals or amines, such as alkali metals and alkaline earth metal hydroxides or organic amines. Examples of metals used as cations are sodium, potassium, magnesium, calcium, etc. Examples of suitable amines are N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine and procaine.

[0074] Base addition salts of acidic compounds may be prepared by contacting the free acid form with a sufficient amount of the desired base to form the salt in a conventional manner. The free acid may be regenerated by contacting the salt form with an acid and isolating the free acid in a conventional manner. The free acid forms differ from their respective salt forms somewhat in certain physical properties, such as solubility in polar solvents, but for purposes of the present invention, the salts are equivalent to their respective free acids.

[0075] Salts can be sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides prepared from inorganic acids, such as hydrochloric acid, nitric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, etc. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, toluenesulfonate, citrate, maleate, fumarate, succinate, tartrate, naphthoate, methanesulfonate, glucoheptonate, lactobionate, laurylsulfonate and isethionate, etc. Salts can also be prepared from organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Representative salts include acetate, propionate, octanoate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, naphthoate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartrate, methanesulfonate, etc. Pharmaceutically acceptable salts may include cations based on alkali and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium, etc., as well as non-toxic ammonium, quaternary ammonium, and amine cations, including but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc. Also contemplated are salts of amino acids, such as argininate, gluconate, galacturonate, and the like (see, e.g., Berge SM et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977; 66: 1-19, incorporated herein by reference).

[0076] "Subjects" for administration include, but are not limited to, humans (i.e., males or females of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or older adults)) and / or non-human animals, e.g., mammals, e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats, and / or dogs. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human animal. The terms "human," "patient," and "subject" are used interchangeably herein.

[0077] "Disease," "disorder," and "condition" are used interchangeably herein.

[0078] As used herein, and unless otherwise indicated, the terms "treatment," "treating," "treating," "treating," and "treatment" include actions that occur while a subject has a particular disease, disorder, or condition, that reduce the severity of, or delay or slow the development of, the disease, disorder, or condition ("therapeutic treatment"), and also include actions that occur before a subject develops a particular disease, disorder, or condition ("prophylactic treatment").

[0079] Generally, the "effective amount" of a compound refers to an amount sufficient to cause a target biological response. As will be appreciated by those of ordinary skill in the art, the effective amount of the compounds of the invention may vary depending on factors such as the biological target, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, health, and symptoms of the subject. An effective amount includes a therapeutically effective amount and a prophylactically effective amount.

[0080] Unless otherwise specified, a "therapeutically effective amount" of a compound as used herein is an amount sufficient to provide a therapeutic benefit in the treatment of a disease, disorder, or condition, or to delay or minimize one or more symptoms associated with the disease, disorder, or condition. A therapeutically effective amount of a compound refers to an amount of a therapeutic agent, alone or in combination with other therapies, that provides a therapeutic benefit in the treatment of a disease, disorder, or condition. The term "therapeutically effective amount" can include an amount that improves overall treatment, reduces or avoids symptoms or causes of a disease or condition, or enhances the therapeutic effect of other therapeutic agents.

[0081] Unless otherwise specified, a "prophylactically effective amount" of a compound as used herein is an amount sufficient to prevent a disease, disorder, or condition, or an amount sufficient to prevent one or more symptoms associated with a disease, disorder, or condition, or an amount to prevent the recurrence of a disease, disorder, or condition. A prophylactically effective amount of a compound refers to an amount of a therapeutic agent that provides a prophylactic benefit in the process of preventing a disease, disorder, or condition when used alone or in combination with other agents. The term "prophylactically effective amount" may include an amount that improves overall prevention, or an amount that enhances the prophylactic effect of other prophylactic agents.

[0082] "Combination" and related terms refer to the simultaneous or sequential administration of the compounds of the invention and other therapeutic agents. For example, the compounds of the invention can be administered simultaneously or sequentially with the other therapeutic agents in separate unit dosage forms, or can be administered simultaneously with the other therapeutic agents in a single unit dosage form. DETAILED DESCRIPTION

[0083] The embodiments of the present invention will be further described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. If no specific conditions are specified in the examples, the conditions are carried out according to normal conditions or manufacturer recommendations. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be obtained commercially.

[0084] Herein, "compounds of the present invention" refers to compounds of the following formula (I), formula (II), etc., and pharmaceutically acceptable salts, isotopic variants, tautomers, stereoisomers, prodrugs, polymorphs, hydrates or solvates thereof; "conjugates of the present invention" are conjugates of the following formula (III), etc.

[0085] For compounds with asymmetric centers, it should be understood that (unless otherwise specified) all optical isomers and mixtures thereof are included. In addition, unless otherwise specified, all isomeric compounds included in the present invention may appear in the form of Z and E with carbon-carbon double bonds. Compounds that exist in different tautomeric forms, a compound is not limited to any specific tautomer, but is intended to cover all tautomeric forms.

[0086] In one embodiment, the present invention relates to a compound of formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0087]

[0088] in,

[0089] M1 is independently selected from -O-, -S-, -NR6-, -CHR7-, -C(O)-, -S(O)- and -S(O)2-;

[0090] R1 is an asialoglycoprotein receptor (ASGPR) ligand residue;

[0091] R2 is selected from hydroxyl, carboxyl, -OR p 、-OPG2、

[0092] PG1 is a carboxyl protecting group;

[0093] PG2 is a hydroxyl protecting group;

[0094] R p for

[0095] PG3 is independently selected from phosphate protecting groups;

[0096] R and R' are each independently an amino protecting group; or R, R' and the N atom to which they are attached optionally together form a 5-10 membered heterocyclic group;

[0097] R3 is independently

[0098] M 3aIndependently selected from chemical bonds, -O-, -S-, -NR6-, -CHR7-, -C(O)-, -S(O) 1-2 -, -NR6C(O)-, -C(O)NR6-, -NR6S(O) 1-2 -、-S(O) 1-2 NR6-, -OC(O)-, -C(O)O-, -OS(O) 1-2 -and-S(O) 1-2 O-;

[0099] M 3b Independently selected from -O-, -S-, -NR6-, -CHR7-, -CH=CH-, -C≡C-, -C(O)-, -S(O) 1-2 -, -C(O)NR6-, -NR6C(O)-, -OC(O)-, -C(O)O-, -S(O) 1-2 O-Y1-、-OS(O) 1-2 -Y1-, -OS(O) 1-2 O-、-S(O) 1- 2NR6-Y1-、-NR6S(O) 1-2 -Y1-, C 6-10 Arylene, 5-10 membered heteroarylene,

[0100] Y1 is independently C 6-10 Arylene or 5-10 membered heteroarylene;

[0101] R6 are each independently selected from H, C 1-10 Alkyl, C 1-10 Haloalkyl, C 3-10 Cycloalkyl and 3-10 membered heterocyclic group;

[0102] R7 are each independently selected from H, halogen, CN, NO2, C 1-10 Alkyl, C 1-10 Haloalkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl and 5-10 membered heteroaryl, wherein one or more methylene groups in the alkyl or haloalkyl are optionally and independently replaced by a group selected from the following: -C(O)-, -C(O)NH-, -NHC(O)-, -OC(O)-, -O-, -C(O)O-, -S-, -S-(CH2) 0-6 -S-、-NR6-、C 6-10 Arylene, 5-10 membered heteroarylene, -CH=CH-, -C≡C- and -O-(CH2) 1-6 -O-;

[0103] n2 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0104] n3 is independently 1, 2, 3, 4, 5 or 6;

[0105] R4 is selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl and C 1-6 Alkoxy;

[0106] Link1, Link2, Link3 and Link4 are each independently C 1-25 Straight chain alkylene, wherein one or more methylene groups in the straight chain alkylene are optionally and independently replaced by R*, and R* is independently selected from: -O-, -S-, -NR6-, -CHR7-, -C(O)-, -S(O) 1-2 -, -NR6C(O)-, -C(O)NR6-, -NR6S(O) 1-2 -、-S(O) 1-2 NR6-, -OC(O)-, -C(O)O-, -OS(O) 1-2 -and-S(O) 1-2 O-;

[0107] m1, m2 and m3 are each independently 1, 2, 3, 4 or 5;

[0108] The condition is that the methylene group connected to the carbonyl group on its left side in Link4 is not replaced by R*.

[0109] M1

[0110] In one embodiment, M1 is -O-; in another embodiment, M1 is -S-; in another embodiment, M1 is -NR6-, such as -NH-; in another embodiment, M1 is -CHR7-, such as -CH2-; in another embodiment, M1 is -C(O)-; in another embodiment, M1 is -S(O)-; in another embodiment, M1 is -S(O)2-;

[0111] In a specific embodiment, M1 is independently selected from -O-, -S-, -NH-, -CH2-, -C(O)-, -S(O)- and -S(O)2-; in another more specific embodiment, M1 is independently selected from -O-, -S- and -NH-; in another more specific embodiment, M1 is -O-.

[0112] R1

[0113] In one embodiment, R1 is an asialoglycoprotein receptor ligand residue; in another embodiment, the asialoglycoprotein receptor ligand is D-mannopyranose; in another embodiment, the asialoglycoprotein receptor ligand is L-mannopyranose; in another embodiment, the asialoglycoprotein receptor ligand is L-arabinose; in another embodiment, the asialoglycoprotein receptor ligand is D-arabinose; in another embodiment, the asialoglycoprotein receptor ligand is D-xylofuranose; in another embodiment, the asialoglycoprotein receptor ligand is L-xylofuranose; in another embodiment, the asialoglycoprotein receptor ligand is D-glucose; in another embodiment In one embodiment, the asialoglycoprotein receptor ligand is L-glucose; in another embodiment, the asialoglycoprotein receptor ligand is D-galactose; in another embodiment, the asialoglycoprotein receptor ligand is L-galactose; in another embodiment, the asialoglycoprotein receptor ligand is α-D-mannofuranose; in another embodiment, the asialoglycoprotein receptor ligand is β-D-mannofuranose; in another embodiment, the asialoglycoprotein receptor ligand is α-D-mannopyranose; in another embodiment, the asialoglycoprotein receptor ligand is β-D-mannopyranose; in another embodiment, the asialoglycoprotein receptor ligand is α- In another embodiment, the asialoglycoprotein receptor ligand is β-D-glucopyranose; in another embodiment, the asialoglycoprotein receptor ligand is α-D-glucopyranose; in another embodiment, the asialoglycoprotein receptor ligand is β-D-glucopyranose; in another embodiment, the asialoglycoprotein receptor ligand is β-D-glucopyranose; in another embodiment, the asialoglycoprotein receptor ligand is α-D-fructofuranose; in another embodiment, the asialoglycoprotein receptor ligand is β-D-fructofuranose; in another embodiment, the asialoglycoprotein receptor ligand is α-D-fructopyranose; in another embodiment, the asialoglycoprotein receptor ligand is β-D-fructopyranose; in another embodiment In one embodiment, the asialoglycoprotein receptor ligand is α-D-galactopyranose; in another embodiment, the asialoglycoprotein receptor ligand is β-D-galactopyranose; in another embodiment, the asialoglycoprotein receptor ligand is α-D-galactofuranose; in another embodiment, the asialoglycoprotein receptor ligand is β-D-galactofuranose; in another embodiment, the asialoglycoprotein receptor ligand is glucosamine; in another embodiment, the asialoglycoprotein receptor ligand is sialic acid; in another embodiment, the asialoglycoprotein receptor ligand is galactosamine; in another embodiment, the asialoglycoprotein receptor ligand is N-acetylgalactosamine;In another embodiment, the asialoglycoprotein receptor ligand is N-trifluoroacetylgalactosamine; in another embodiment, the asialoglycoprotein receptor ligand is N-propionylgalactosamine; in another embodiment, the asialoglycoprotein receptor ligand is N-n-butyrylgalactosamine; in another embodiment, the asialoglycoprotein receptor ligand is N-isobutyrylgalactosamine; in another embodiment, the asialoglycoprotein receptor ligand is 2-amino-3-O-[(R)-1-carboxyethyl]-2-deoxy-β-D-pyranose. Sugar; in another embodiment, the asialoglycoprotein receptor ligand is 2-deoxy-2-methylamino-L-pyranose; in another embodiment, the asialoglycoprotein receptor ligand is 4,6-dideoxy-4-formamido-2,3-di-O-methyl-D-pyranose; in another embodiment, the asialoglycoprotein receptor ligand is 2-deoxy-2-sulfoamino-D-pyranose; in another embodiment, the asialoglycoprotein receptor ligand is N-glycolyl-α-neuraminic acid; in another embodiment , the asialoglycoprotein receptor ligand is 5-thio-β-D-glucopyranose; in another embodiment, the asialoglycoprotein receptor ligand is 2,3,4-tri-O-acetyl-1-thio-6-O-trityl-α-D-glucopyranoside methyl ester; in another embodiment, the asialoglycoprotein receptor ligand is 4-thio-β-D-galactopyranose; in another embodiment, the asialoglycoprotein receptor ligand is 3,4,6,7-tetra-O-acetyl-2-deoxy-1,5-dithio-α-D-glucopyranoside Heptose ethyl ester; in another embodiment, the asialoglycoprotein receptor ligand is 2,5-anhydro-D-allose nitrile; in another embodiment, the asialoglycoprotein receptor ligand is ribose; in another embodiment, the asialoglycoprotein receptor ligand is D-ribose; in another embodiment, the asialoglycoprotein receptor ligand is D-4-thioribose; in another embodiment, the asialoglycoprotein receptor ligand is L-ribose; in another embodiment, the asialoglycoprotein receptor ligand is L-4-thioribose. ;

[0114] In a specific embodiment, R1 is independently selected from the residues of the following structures: D-pyranose mannosyl; L-pyranose mannosyl; L-arabinose; D-arabinose; D-xylofuranose; L-xylofuranose; D-glucose; L-glucose; D-galactose; L-galactose; α-D-furanose mannosyl; β-D-furanose mannosyl; β-D-pyranose mannosyl; β-D-pyranose mannosyl; α-D-pyranose glucose; β-D -Glucopyranose, α-D-glucofuranose, β-D-glucofuranose, α-D-fructofuranose, β-D-fructofuranose, α-D-fructopyranose, β-D-fructopyranose, α-D-galactopyranose, β-D-galactopyranose, α-D-galactofuranose, β-D-galactofuranose, glucosamine, sialic acid, galactosamine, N-acetylgalactosamine, N-trifluoroacetylgalactosamine, N-propionylgalactosamine, N-butylgalactosamine acylgalactosamine, N-isobutyrylgalactosamine, 2-amino-3-O-[(R)-1-carboxyethyl]-2-deoxy-β-D-glucopyranose, 2-deoxy-2-methylamino-L-glucopyranose, 4,6-dideoxy-4-formamido-2,3-di-O-methyl-D-mannopyranose, 2-deoxy-2-sulfoamino-D-glucopyranose, N-glycolyl-α-neuraminic acid, 5-thio-β-D- Glucopyranose, 2,3,4-tri-O-acetyl-1-thio-6-O-trityl-α-D-glucopyranoside methyl ester, 4-thio-β-D-galactopyranose, 3,4,6,7-tetra-O-acetyl-2-deoxy-1,5-dithio-α-D-glucopyranose heptanoside ethyl ester, 2,5-anhydro-D-allose nitrile, ribose, D-ribose, D-4-thioribose, L-ribose and L-4-thioribose.

[0115] In another specific embodiment, R1 is independently selected from the residues of the following structures: D-galactose, L-galactose, α-D-galactopyranose, β-D-galactopyranose, α-D-galactofuranose, β-D-galactofuranose, galactosamine, N-acetylgalactosamine, N-trifluoroacetylgalactosamine, N-propionylgalactosamine, N-butyrylgalactosamine, N-isobutyrylgalactosamine and β-D-galactopyranose.

[0116] In another specific embodiment, R1 is independently selected from the residues of the following structures: galactosamine, N-acetylgalactosamine, N-trifluoroacetylgalactosamine, N-propionylgalactosamine, N-n-butyrylgalactosamine and N-isobutyrylgalactosamine.

[0117] In another specific embodiment, R1 is an N-acetylgalactosamine residue.

[0118] In another specific embodiment, R1 is

[0119] R2

[0120] In one embodiment, R2 is hydroxyl; in another embodiment, R2 is carboxyl; in another embodiment, R2 is -OR p ,For example In another embodiment, R2 is -OPG2; in another embodiment, R2 is For example For example In another embodiment, R2 is For example For example For example For example For example For example

[0121] In a specific embodiment, R2 is selected from hydroxyl, carboxyl, -OR p 、-OPG2、 In another specific embodiment, R2 is selected from -OR p , In another specific embodiment, R2 is selected from In another specific embodiment, R2 is selected from In another specific embodiment, R2 is selected from In another specific embodiment, R2 is In another specific embodiment, R2 is selected from In another specific embodiment, R2 is In another specific embodiment, for In another specific embodiment, for In another specific embodiment, for

[0122] PG1

[0123] In one embodiment, PG1 is a carboxyl protecting group, such as C 1-6 Alkyl, such as methyl, such as tert-butyl, such as benzyl, such as allyl, such as For example For example

[0124] In a specific embodiment, PG1 is selected from C 16 Alkyl, benzyl, allyl, In another more specific embodiment, PG1 is selected from methyl, tert-butyl, benzyl, allyl, In another more specific embodiment, PG1 is selected from In another more specific embodiment, PG1 is In another more specific embodiment, PG1 is

[0125] In one embodiment, R 2s is H; in another embodiment, R 2s is halogen, such as F; in another embodiment, R 2s C 1-10 Alkyl groups, such as C 1-6 Alkyl; in another embodiment, R 2s C 1-10 Haloalkyl, such as C 1-6 Halogenated alkyl.

[0126] In a specific embodiment, R 2s are independently selected from H, halogen, C 1-10 Alkyl and C 1-10 In another specific embodiment, R 2s are independently selected from H, halogen, C 1-6 Alkyl and C 1-6 In another specific embodiment, R 2s is H or halogen; in another specific embodiment, R 2s For F.

[0127] In one embodiment, q is 0; in another embodiment, q is 1; in another embodiment, q is 2; in another embodiment, q is 3; in another embodiment, q is 4; in another embodiment, q is 5.

[0128] PG2

[0129] In one embodiment, PG2 is a hydroxy protecting group, such as trimethylsilyl (TMS), such as triethylsilyl (TES), such as dimethylisopropylsilyl (DMIPS), such as diethylisopropylsilyl (DEIPS), such as tert-butyldimethylsilyl ether (TBDMS), such as tert-butyldiphenylsilyl (TBDPS), such as triisopropylsilyl ether (TIPS), such as acetyl (Ac), such as chloroacetyl, such as dichloroacetyl, such as trichloroacetyl, such as trifluoroacetyl (TFA), such as benzoyl, such as p-methoxybenzoyl, such as 9-fluorenylmethoxycarbonyl (Fmoc), such as allyloxycarbonyl (Alloc), such as 2,2,2-trichloroethoxycarbonyl (Troc), such as benzyloxycarbonyl (Cbz), such as tert-butyloxycarbonyl (B oc), for example benzyl (Bn), for example p-methoxybenzyl (PMB), for example allyl, for example dimethoxytrityl (DMT), for example monomethoxytrityl (MMT), for example 9-phenylxanthen-9-yl (Pixyl), for example 9-(p-methoxyphenyl)xanthen-9-yl (Mox), for example trityl (Tr), for example 4-methoxytrityl (MMTr), for example 4,4'-dimethoxytrityl (DMTr), for example 4,4',4"-trimethoxytrityl (TMTr), for example methoxymethyl (MOM), for example phenoxymethyl (BOM), for example 2,2,2-trichloroethoxymethyl, for example 2-methoxyethoxymethyl (MEM), for example methylthiomethyl (MTM), for example p-methoxybenzyloxymethyl (PMBM).

[0130] In a specific embodiment, PG2 is selected from: trimethylsilyl (TMS), triethylsilyl (TES), dimethylisopropylsilyl (DMIPS), diethylisopropylsilyl (DEIPS), tert-butyldimethylsilyl ether (TBDMS), tert-butyldiphenylsilyl (TBDPS), triisopropylsilyl ether (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, dimethoxytrityl (DMT), monomethoxytrityl (MMT), 9-phenylxanthen-9-yl (Pixyl), 9-(p-methoxyphenyl)xanthen-9-yl (Mox), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr), 4,4',4"-trimethoxytrityl (TMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), and p-methoxybenzyloxymethyl (PMBM).

[0131] In another specific embodiment, PG2 is selected from: dimethoxytrityl (DMT), monomethoxytrityl (MMT), 9-phenylxanthen-9-yl (Pixyl), 9-(p-methoxyphenyl)xanthen-9-yl (Mox), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr), 4,4',4"-trimethoxytrityl (TMTr), tert-butyldimethylsilyl ether (TBMDS), tert-butyldiphenylsilyl (TBDPS) and triisopropylsilyl ether (TIPS).

[0132] In another specific embodiment, PG2 is selected from the group consisting of dimethoxytrityl (DMT), monomethoxytrityl (MMT), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr) and 4,4',4"-trimethoxytrityl (TMTr).

[0133] In another specific embodiment, PG2 is 4,4'-dimethoxytrityl (DMTr).

[0134] R p

[0135] In one embodiment, Rp for For example

[0136] PG3

[0137] In one embodiment, PG3 is a phosphate protecting group, such as 2-cyanoethyl, such as C 1-6 An alkyl group, such as a methyl group.

[0138] In a more specific embodiment, PG3 is independently selected from 2-cyanoethyl and C 1-6 Alkyl; in a more specific embodiment, PG3 is independently selected from 2-cyanoethyl and methyl; in a more specific embodiment, PG3 is 2-cyanoethyl.

[0139] R and R'

[0140] In one embodiment, R and R' are each independently an amino protecting group, such as C 1-6 Alkyl, such as isopropyl, such as R, R' and the nitrogen atom to which they are attached, are optionally taken together to form a 5-6 membered heterocyclic group.

[0141] R3

[0142] In one embodiment, R3 is independently In another embodiment, R3 is In another embodiment, R3 is

[0143] In one embodiment, is A1; in another embodiment, A2; in another embodiment, A3; in another embodiment, A4; in another embodiment, A5; in another embodiment, A6; in another embodiment, is A7; in another embodiment, is A8; in another embodiment, is A9; in another embodiment, is A10; in another embodiment, is A11; in another embodiment, is A12; in another embodiment, is A13; in another embodiment, is A14; in another embodiment, is A15; in another embodiment, is A16; in another embodiment, is A17; in another embodiment, is A18; in another embodiment, is A19; in another embodiment, is A20; in another embodiment, A21; in another embodiment, is A22; in another embodiment, A23; in another embodiment, For A24.

[0144] The structure of A1-A24 is as follows:

[0145]

[0146] In a specific embodiment, is independently selected from the structures in A1-A24; in another more specific embodiment, are independently selected from A2, A3, A4, A6 and A9; in another more specific embodiment, are independently selected from A2, A3 and A9; in another more specific embodiment, Independently A3 or A9.

[0147] M 3a

[0148] In one embodiment, M 3a is a chemical bond; in another embodiment, M 3a is -O-; in another embodiment, M 3a is -S-; in another embodiment, M 3a is -NR6-, such as -NH-; in another embodiment, M 3a is -CHR7-, for example -CH2-; in another embodiment, M 3a is -C(O)-; in another embodiment, M 3a -S(O) 1-2 -; In another embodiment, M 3a is -NR6C(O)-, for example -NHC(O)-; in another embodiment, M 3a is -C(O)NR6-, for example -C(O)NH-; in another embodiment, M 3a -NR6S(O) 1-2 -; In another embodiment, M 3a -S(O) 1-2NR6-; In another embodiment, M 3a is -OC(O)-; in another embodiment, M 3a is -C(O)O-; in another embodiment, M 3a For-OS(O) 1-2 -; In another embodiment, M 3a -S(O) 1-2 O-.

[0149] In a specific embodiment, M 3a Independently selected from chemical bonds, -O-, -S-, -NR6-, -CHR7-, -C(O)-, -S(O) 1-2 -, -NHC(O)-, -C(O)NH-, -NHS(O) 1-2 -、-S(O) 1-2 NH-, -OC(O)-, -C(O)O-, -OS(O) 1-2 -and-S(O) 1-2 O-; In another specific embodiment, M 3a is independently selected from a chemical bond, -O-, -S-, -NR6-, -C(O)-, -NHC(O)-, -C(O)NH-, -OC(O)-, and -C(O)O-; in another specific embodiment, M 3a is independently selected from a chemical bond, -NHC(O)-, -C(O)NH-, -OC(O)-, and -C(O)O-; in another specific embodiment, M 3a is independently selected from a chemical bond, -NHC(O)- and -C(O)NH-; in another specific embodiment, M 3a are independently a chemical bond or -C(O)NH-.

[0150] M 3b

[0151] In one embodiment, M 3b is -O-; in another embodiment, M 3b is -S-; in another embodiment, M 3b is -NR6-; in another embodiment, M 3b is -CHR7-; in another embodiment, M 3b is -CH=CH-; in another embodiment, M 3b is -C≡C-; in another embodiment, M 3b is -C(O)-; in another embodiment, M 3b -S(O) 1-2 -; In another embodiment, M 3bis -C(O)NR6-, for example -C(O)NH-; in another embodiment, M 3b is -NR6C(O)-, for example -NHC(O)-; in another embodiment, M 3b is -OC(O)-; in another embodiment, M 3b is -C(O)O-; in another embodiment, M 3b -S(O) 1-2 O-Y1-; In another embodiment, M 3b For-OS(O) 1-2 -Y1-; In another embodiment, M 3b For-OS(O) 1-2 O-; In another embodiment, M 3b -S(O) 1-2 NR6-Y1-; In another embodiment, M 3b -NR6S(O) 1-2 -Y1-; In another embodiment, M 3b C 6-10 Arylene, such as phenylene; in another embodiment, M 3b is a 5-10 membered heteroarylene group, for example a 5-6 membered heteroarylene group, for example For example In another embodiment, M 3b for In another embodiment, M 3b for In another embodiment, M 3b for

[0152] In a specific embodiment, M 3b Independently selected from -O-, -S-, -NR6-, -CHR7-, -CH=CH-, -C≡C-, -C(O)-, -S(O) 1-2 -, -C(O)NH-, -NHC(O)-, -OC(O)-, -C(O)O-, -S(O) 1-2 O-Y1-、-OS(O) 1-2 -Y1-, -OS(O) 1-2 O-、-S(O) 1-2 NH-Y1-, -NHS(O) 1-2 -Y1-, phenylene, 5-6 membered heteroarylene, In another specific embodiment, M 3bindependently selected from -O-, -S-, -NR6-, -CHR7-, -CH=CH-, -C≡C-, -C(O)-, -C(O)NH-, -NHC(O)-, -OC(O)-, -C(O)O-, -OS(O)2-Y1-, -OS(O)2O-, -NHS(O)2-Y1-, 5-6 membered heteroarylene, In another specific embodiment, M 3b are independently selected from -C(O)NH-, -NHC(O)-, -OC(O)-, -C(O)O-; in another specific embodiment, M 3b is -C(O)NH- or -NHC(O)-; in another specific embodiment, M 3b is -NHC(O)-; in another specific embodiment, M 3b The 5-6 membered heteroarylene group is

[0153] Y1

[0154] In one embodiment, Y1 is C 6-10 In another embodiment, Y1 is a 5-10 membered heteroarylene group, such as a 5-6 membered heteroarylene group.

[0155] In one specific embodiment, Y1 is independently phenylene or 5-6 membered heteroarylene; in another more specific embodiment, Y1 is phenylene, for example

[0156] R6

[0157] In one embodiment, R6 is H; in another embodiment, R6 is C 1-10 Alkyl groups, such as C 1-6 Alkyl, such as -CH3, such as For example For example In another embodiment, R6 is C 1-10 Haloalkyl, such as C 1-6 haloalkyl; in another embodiment, R6 is C 3-10 Cycloalkyl, such as C 3-7 In another embodiment, R6 is a 3-10 membered heterocyclyl, such as a 3-7 membered heterocyclyl.

[0158] In a more specific embodiment, R6 are each independently selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-7 Cycloalkyl and 3-7 membered heterocyclyl; In another more specific embodiment, R6 are each independently selected from H, C1-6 Alkyl and C 1-6 In another more specific embodiment, R6 are each independently selected from H, -CH3, In another more specific embodiment, R6 is H.

[0159] R7

[0160] In one embodiment, R7 is H; in another embodiment, R7 is halogen, such as F, such as Cl, such as Br; in another embodiment, R7 is CN; in another embodiment, R7 is NO2; in another embodiment, R7 is C 1-10 Alkyl; in another embodiment, R7 is C 1-10 haloalkyl; in another embodiment, R7 is C 3-10 In another embodiment, R7 is a 3-10 membered heterocyclyl; In another embodiment, R7 is C 6-10 Aryl; in another embodiment, R7 is a 5-10 membered heteroaryl.

[0161] In one embodiment, one or more methylene groups in the alkyl or haloalkyl in R7 are optionally and independently replaced by a group selected from the following: -C(O)-, -C(O)NH-, -NHC(O)-, -OC(O)-, -O-, -C(O)O-, -S-, -S-(CH2) 0-6 -S-、-NR6-、C 6-10 Arylene, 5-10 membered heteroarylene, -CH=CH-, -C≡C- and -O-(CH2) 1-6 -O-; for example substituted by -C(O)-, for example substituted by -C(O)NH-, for example substituted by -NHC(O)-, for example substituted by -OC(O)-, for example substituted by -O-, for example substituted by -C(O)O-, for example substituted by -S-, for example substituted by -S-(CH2) 0-6 -S- is replaced, for example, by -NR6-, for example, by C 6-10 Arylene is replaced, for example, by a 5-10 membered heteroarylene, for example, by -CH=CH-, for example, by -C≡C-, for example, by -O-(CH2) 1-6 -O-substituted; In another embodiment, 1, 2 or 3 methylene groups in the alkyl or haloalkyl group in R7 are optionally and independently replaced by groups selected from the foregoing.

[0162] In a more specific embodiment, R7 are each independently selected from H, halogen, CN, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-7Cycloalkyl, 3-7 membered heterocyclyl, phenyl and 5-6 membered heteroaryl; In another more specific embodiment, R7 is each independently selected from H, halogen, CN, C 1-6 Alkyl and C 1-6 In another more specific embodiment, R7 are each independently selected from H, halogen, C 1-6 Alkyl and C 1-6 Halogenated alkyl.

[0163] In a more specific embodiment, one or more methylene groups in the alkyl or haloalkyl in R7 are optionally and independently replaced by a group selected from the following: -C(O)-, -C(O)NH-, -NHC(O)-, -OC(O)-, -O-, -C(O)O-, -S-, -S-(CH2) 0-3 -S-, -NR6-, phenylene, 5-6 membered heteroarylene, -CH=CH-, -C≡C- and -O-(CH2) 1-3 In another more specific embodiment, 1, 2 or 3 methylene groups in the alkyl or haloalkyl group in R7 are optionally and independently replaced by groups selected from the following: -C(O)-, -C(O)NH-, -NHC(O)-, -OC(O)-, -O-, -C(O)O-, -S-, -SS-, -NR6-, -CH=CH-, -C≡C- and -O-(CH2)2-O-; in another more specific embodiment, the halogen in R7 is -F, -Cl or -Br.

[0164] n2

[0165] In one embodiment, n2 is 0; in another embodiment, n2 is 1; in another embodiment, n2 is 2; in another embodiment, n2 is 3; in another embodiment, n2 is 4; in another embodiment, n2 is 5; in another embodiment, n2 is 6; in another embodiment, n2 is 7; in another embodiment, n2 is 8; in another embodiment, n2 is 9; in another embodiment, n2 is 10.

[0166] n3

[0167] In one embodiment, n3 is 1; in another embodiment, n3 is 2; in another embodiment, n3 is 3; in another embodiment, n3 is 4; in another embodiment, n3 is 5; in another embodiment, n3 is 6.

[0168] R4

[0169] In one embodiment, R4 is H; in another embodiment, R4 is C1-6 Alkyl groups, such as C 1-5 Alkyl groups, such as C 1-3 Alkyl; in another embodiment, R4 is C 1-6 Haloalkyl, such as C 1-5 Haloalkyl, such as C 1-3 haloalkyl; in another embodiment, R4 is C 1-6 Alkoxy.

[0170] In a more specific embodiment, R4 is selected from H, C 1-5 Alkyl, C 1-5 Haloalkyl and C 1-5 Alkoxy; in another more specific embodiment, R4 is selected from H, C 1-3 Alkyl and C 1-3 In another more specific embodiment, R4 is H.

[0171] Link1, Link2 and Link3

[0172] In one embodiment, Link1 is C 1-25 Straight chain alkylene, such as C 1-10 Straight chain alkylene, such as C 1-6 Straight chain alkylene, such as C 4-6 Straight chain alkylene, for example -(CH2)4-, for example -(CH2)6-.

[0173] In one embodiment, Link2 is C 1-25 Straight chain alkylene, such as C 1-10 Straight chain alkylene, such as C 1-6 Straight chain alkylene, such as C 4-6 Straight chain alkylene, for example -(CH2)4-, for example -(CH2)6-.

[0174] In one embodiment, Link3 is C 1-25 Straight chain alkylene, such as C 1-10 Straight chain alkylene, such as C 1-6 Straight chain alkylene, such as C 4-6 Straight chain alkylene, for example -(CH2)4-, for example -(CH2)6-.

[0175] In one embodiment, one or more methylene groups in Link1, Link2 and Link3 are optionally and independently replaced by R*; in another embodiment, 1, 2 or 3 methylene groups in Link1, Link2 and Link3 are optionally and independently replaced by R*; in another embodiment, 1 methylene group in Link1, Link2 and Link3 is optionally and independently replaced by R*.

[0176] In a more specific embodiment, Link1, Link2 and Link3 are each independently C 1-10 In another more specific embodiment, Link1, Link2 and Link3 are each independently C 1-6 In another more specific embodiment, Link1, Link2 and Link3 are each independently C 4-6 Straight chain alkylene; in another more specific embodiment, Link1, Link2 and Link3 are each independently -(CH2)4- or -(CH2)6-.

[0177] In one embodiment, Link1, Link2 and Link3 are each independently

[0178] In one embodiment, n1 is 1; in another embodiment, n1 is 2; in another embodiment, n1 is 3; in another embodiment, n1 is 4; in another embodiment, n1 is 5; in another embodiment, n1 is 6; in another embodiment, n1 is 7; in another embodiment, n1 is 8; in another embodiment, n1 is 9; in another embodiment, n1 is 10.

[0179] Link4

[0180] In one embodiment, Link4 is C 1-25 Straight chain alkylene, such as C 5-25 Straight chain alkylene, such as C 1-20 Straight chain alkylene, such as C 5-20 Straight chain alkylene, such as C 8-20 Straight chain alkylene, such as C 1-17 Straight chain alkylene, such as C 10-17 Straight chain alkylene, such as C 1-10 Straight chain alkylene, such as C 3-20 Straight chain alkylene, such as C 3-17 Straight chain alkylene, such as C 3-10 Straight chain alkylene, such as -(CH2)3-, such as -(CH2) 10 -.

[0181] In one embodiment, one or more methylene groups in Link4 are optionally and independently replaced by R*; in another embodiment, 1, 2 or 3 methylene groups in Link4 are optionally and independently replaced by R*; in another embodiment, 1 methylene group in Link4 is optionally and independently replaced by R*, for example, Link4 is -(CH2)3-C(O)NH-(CH2)6-, for example, Link4 is -(CH2) 10 -C(O)NH-(CH2)6-; In another embodiment, the methylene group attached to the carbonyl group on its left side in Link4 is not replaced by R*.

[0182] In a more specific embodiment, Link4 is selected from -(CH2)3-, -(CH2) 10 -, -(CH2)3-C(O)NH-(CH2)6- and -(CH2) 10 -C(O)NH-(CH2)6-; In another more specific embodiment, Link4 is selected from -(CH2)3-, -(CH2) 10 - and -(CH2)3-C(O)NH-(CH2)6-; in another more specific embodiment, Link4 is -(CH2)3- or -(CH2) 10 -; In another more specific embodiment, Link4 is selected from -(CH2) 10 -, -(CH2)3-C(O)NH-(CH2)6- and -(CH2) 10 -C(O)NH-(CH2)6-; In another more specific embodiment, Link4 is -(CH2) 10 -or-(CH2) 10 -C(O)NH-(CH2)6-.

[0183] R*

[0184] In one embodiment, R* is -O-; in another embodiment, R* is -S-; in another embodiment, R* is -NR6-; in another embodiment, R* is -CHR7-; in another embodiment, R* is -C(O)-; in another embodiment, R* is -S(O) 1-2 -; in another embodiment, R* is -NR6C(O)-, such as -NHC(O)-; in another embodiment, R* is -C(O)NR6-, such as -C(O)NH-; in another embodiment, R* is -NR6S(O) 1-2 -; In another embodiment, R* is -S(O) 1-2NR6-; in another embodiment, R* is -OC(O)-; in another embodiment, R* is -C(O)O-; in another embodiment, R* is -OS(O) 1-2 -; In another embodiment, R* is -S(O) 1-2 O-.

[0185] In a more specific embodiment, R* is independently selected from: -O-, -NR6-, -CHR7-, -C(O)-, -NR6C(O)-, -C(O)NR6-, -OC(O)- and -C(O)O-; in another more specific embodiment, R* is independently selected from: -C(O)NH-, -NHC(O)-, -C(O)O- and -OC(O)-; in another more specific embodiment, R* is independently selected from: -C(O)NH- and -C(O)O-; in another more specific embodiment, R* is independently selected from: -C(O)NH- and -NHC(O)-; in another more specific embodiment, R* is independently selected from: -C(O)NH- and -NHC(O)-; in another more specific embodiment, R* is -C(O)NH-.

[0186] In one embodiment, R* is separated from the carbonyl group on the left side of Link4 by 1-15 methylene groups, for example, by 1 methylene group, for example, by 2 methylene groups, for example, by 3 methylene groups, for example, by 4 methylene groups, for example, by 5 methylene groups, for example, by 6 methylene groups, for example, by 7 methylene groups, for example, by 8 methylene groups, for example, by 9 methylene groups, for example, by 10 methylene groups, for example, by 11 methylene groups, for example, by 12 methylene groups, for example, by 13 methylene groups, for example, by 14 methylene groups, for example, by 15 methylene groups.

[0187] In a more specific embodiment, R* is separated from the carbonyl group on the left side of Link4 by 1-15 methylene groups; in another more specific embodiment, R* is separated from the carbonyl group on the left side of Link4 by 1-12 methylene groups; in another more specific embodiment, R* is separated from the carbonyl group on the left side of Link4 by 1-10 methylene groups; in another more specific embodiment, R* is separated from the carbonyl group on the left side of Link4 by 1-3 methylene groups; in another more specific embodiment, R* is separated from the carbonyl group on the left side of Link4 by 3-10 methylene groups.

[0188] In one embodiment, R* and R2 are separated by 1-12 methylene groups, for example, by 1 methylene group, for example, by 2 methylene groups, for example, by 3 methylene groups, for example, by 4 methylene groups, for example, by 5 methylene groups, for example, by 6 methylene groups, for example, by 7 methylene groups, for example, by 8 methylene groups, for example, by 9 methylene groups, for example, by 10 methylene groups, for example, by 11 methylene groups, for example, by 12 methylene groups.

[0189] In a more specific embodiment, R* is separated from R2 by 1-12 methylene groups; in another more specific embodiment, R* is separated from R2 by 1-10 methylene groups; in another more specific embodiment, R* is separated from R2 by 1-6 methylene groups.

[0190] m1, m2 and m3

[0191] In one embodiment, m1 is 1; in another embodiment, m1 is 2; in another embodiment, m1 is 3; in another embodiment, m1 is 4; in another embodiment, m1 is 5.

[0192] In one embodiment, m2 is 1; in another embodiment, m2 is 2; in another embodiment, m2 is 3; in another embodiment, m2 is 4; in another embodiment, m2 is 5.

[0193] In one embodiment, m3 is 1; in another embodiment, m3 is 2; in another embodiment, m3 is 3; in another embodiment, m3 is 4; in another embodiment, m3 is 5.

[0194] In a more specific embodiment, m1, m2 and m3 are each independently 1, 2 or 3.

[0195] The present invention also provides a conjugate having a structure shown in formula (III):

[0196]

[0197] Wherein, L is a structure represented by formula (I') or formula (II'):

[0198]

[0199] Selected from

[0200] R D is a drug molecule independently selected from a cytotoxic agent, a chemotherapeutic agent, a somatostatin, a toxin, a radioisotope, and an oligonucleotide chain;

[0201] d is 1 or 2, preferably 1;

[0202] The remaining groups are as defined herein.

[0203] L

[0204] In one embodiment, L is a structure represented by formula (I'); in another embodiment, L is a structure represented by formula (II'); in another embodiment, L is L1; in another embodiment, L is L2; ​​in another embodiment, L is L3; in another embodiment, L is L4. The structures of L1, L2, L3 and L4 are as defined herein.

[0205] In a more specific embodiment, L is selected from L1, L2, L3 and L4; in another more specific embodiment, L is selected from L1, L2 and L4.

[0206]

[0207] In one embodiment, for In another embodiment, for For example For example In another embodiment, for For example For example In another embodiment, for For example For example

[0208] In a more specific embodiment, Selected from In another more specific embodiment, Selected from In another more specific embodiment, Selected from Selected from In another more specific embodiment, Selected from Selected from

[0209] R D

[0210] In one embodiment, R D is a drug molecule, such as a cytotoxic agent, such as a chemotherapeutic agent, such as a somatostatin, such as a toxin, such as a radioisotope, such as an oligonucleotide chain.

[0211] In one embodiment, the oligonucleotide chain is a single-stranded oligonucleotide; in another embodiment, the oligonucleotide chain is a double-stranded oligonucleotide.

[0212] In one embodiment, M2 is connected to the 5' end of at least one of the oligonucleotide chains via a phosphate bond; in another embodiment, M2 is connected to the 3' end of at least one of the oligonucleotide chains via a phosphate bond; in another embodiment, M2 is connected to any nucleotide between the 5' end and the 3' end of at least one of the oligonucleotide chains via a phosphate bond; in another embodiment, M2 is connected to the sense strand of the oligonucleotide chain via a phosphate bond; in another embodiment, M2 is connected to the antisense strand of the oligonucleotide chain via a phosphate bond.

[0213] In one embodiment, the phosphate bond is a phosphodiester bond; in another embodiment, the phosphate bond is a modified phosphate bond, such as a thio-modified phosphate bond, such as an amino-modified phosphate bond.

[0214] d

[0215] In one embodiment, d is 1; in another embodiment, d is 2.

[0216] Any technical solution or any combination thereof in any of the above specific embodiments can be combined with any technical solution or any combination thereof in other specific embodiments. For example, any technical solution or any combination thereof of M1 can be combined with R1, R2, PG1, PG2, R p ,PG3,R,R',R3,M1,M2,M 3a 、M 3b , Y1, R6, R7, n1, n2, n3, R4, R D , d, Link1, Link2, Link3, Link4 and L, etc. or any combination thereof. The present invention is intended to include combinations of all these technical solutions, which are not listed one by one due to space limitations.

[0217] In a more specific embodiment, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:

[0218]

[0219] in,

[0220] M1 is independently selected from -O-, -S-, -NR6-, -CHR7-, -C(O)-, -S(O)- and -S(O)2-;

[0221] R1 is an asialoglycoprotein receptor (ASGPR) ligand residue;

[0222] R2 is selected from hydroxyl, carboxyl, -ORp 、-OPG2、

[0223] PG1 is a carboxyl protecting group;

[0224] PG2 is a hydroxyl protecting group;

[0225] R p for

[0226] PG3 is independently selected from phosphate protecting groups;

[0227] R and R' are each independently an amino protecting group; or R, R' and the N atom to which they are attached optionally together form a 5-10 membered heterocyclic group;

[0228] R3 is independently

[0229] M 3a Independently selected from chemical bonds, -O-, -S-, -NR6-, -CHR7-, -C(O)-, -S(O) 1-2 -, -NR6C(O)-, -C(O)NR6-, -NR6S(O) 1-2 -、-S(O) 1-2 NR6-, -OC(O)-, -C(O)O-, -OS(O) 1-2 -and-S(O) 1-2 O-;

[0230] M 3b Independently selected from -O-, -S-, -NR6-, -CHR7-, -CH=CH-, -C≡C-, -C(O)-, -S(O) 1-2 -, -C(O)NR6-, -NR6C(O)-, -OC(O)-, -C(O)O-, -S(O) 1-2 O-Y1-、-OS(O) 1-2 -Y1-, -OS(O) 1-2 O-、-S(O) 1- 2NR6-Y1-、-NR6S(O) 1-2 -Y1-, C 6-10 Arylene, 5-10 membered heteroarylene,

[0231] Y1 is independently C 6-10 Arylene or 5-10 membered heteroarylene;

[0232] R6 are each independently selected from H, C 1-10 Alkyl, C 1-10 Haloalkyl, C 3-10Cycloalkyl and 3-10 membered heterocyclic group;

[0233] R7 are each independently selected from H, halogen, CN, NO2, C 1-10 Alkyl, C 1-10 Haloalkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl and 5-10 membered heteroaryl, wherein one or more methylene groups in the alkyl or haloalkyl are optionally and independently replaced by a group selected from the following: -C(O)-, -C(O)NH-, -NHC(O)-, -OC(O)-, -O-, -C(O)O-, -S-, -S-(CH2) 0-6 -S-、-NR6-、C 6-10 Arylene, 5-10 membered heteroarylene, -CH=CH-, -C≡C- and -O-(CH2) 1-6 -O-;

[0234] n2 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0235] n3 is independently 1, 2, 3, 4, 5 or 6;

[0236] R4 is selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl and C 1-6 Alkoxy;

[0237] Link1, Link2, Link3 and Link4 are each independently C 1-25 Straight chain alkylene, wherein one or more methylene groups in the straight chain alkylene are optionally and independently replaced by R*, and R* is independently selected from: -O-, -S-, -NR6-, -CHR7-, -C(O)-, -S(O) 1-2 -, -NR6C(O)-, -C(O)NR6-, -NR6S(O) 1-2 -、-S(O) 1-2 NR6-, -OC(O)-, -C(O)O-, -OS(O) 1-2 -and-S(O) 1-2 O-;

[0238] m1, m2 and m3 are each independently 1, 2, 3, 4 or 5;

[0239] The condition is that the methylene group connected to the carbonyl group on its left side in Link4 is not replaced by R*.

[0240] In a more specific embodiment, the present invention provides a compound of the above formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein M1 is independently selected from -O-, -S-, -NH-, -CH2-, -C(O)-, -S(O)- and -S(O)2-, preferably selected from -O-, -S- and -NH-, preferably -O-.

[0241] In a more specific embodiment, the present invention provides a compound of formula (I) as described above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R1 is independently selected from the residues of the following structures: D-mannopyranose, L-mannopyranose, L-arabinose, D-arabinose, D-xylofuranose, L-xylofuranose, D-glucose, L-glucose, D-galactose, L-galactose, α-D-mannofuranose, β-D-mannofuranose, α- D-mannopyranose, β-D-mannopyranose, α-D-glucose pyranose, β-D-glucose pyranose, α-D-glucofuranose, β-D-glucofuranose, α-D-fructofuranose, β-D-fructofuranose, α-D-fructopyranose, β-D-fructopyranose, α-D-galactopyranose, β-D-galactopyranose, α-D-galactofuranose, β-D-galactofuranose, glucosamine, sialic acid, galactosamine, N-acetylgalactosamine, N-trimethylol Fluoroacetylgalactosamine, N-propionylgalactosamine, N-butyrylgalactosamine, N-isobutyrylgalactosamine, 2-amino-3-O-[(R)-1-carboxyethyl]-2-deoxy-β-D-glucopyranose, 2-deoxy-2-methylamino-L-glucopyranose, 4,6-dideoxy-4-formamido-2,3-di-O-methyl-D-mannopyranose, 2-deoxy-2-sulfoamino-D-glucopyranose, N-glycolyl-α-neuraminic acid , 5-thio-β-D-glucopyranose, 2,3,4-tri-O-acetyl-1-thio-6-O-trityl-α-D-glucopyranoside methyl ester, 4-thio-β-D-galactopyranose, 3,4,6,7-tetra-O-acetyl-2-deoxy-1,5-dithio-α-D-glucopyranose heptanoside ethyl ester, 2,5-anhydro-D-allose nitrile, ribose, D-ribose, D-4-thioribose, L-ribose and L-4-thioribose.

[0242] Preferably, R1 is independently selected from the residues of the following structures: D-galactose, L-galactose, α-D-galactopyranose, β-D-galactopyranose, α-D-galactofuranose, β-D-galactofuranose, galactosamine, N-acetylgalactosamine, N-trifluoroacetylgalactosamine, N-propionylgalactosamine, N-n-butyrylgalactosamine, N-isobutyrylgalactosamine and β-D-galactopyranose, preferably selected from: galactosamine, N-acetylgalactosamine, N-trifluoroacetylgalactosamine, N-propionylgalactosamine, N-n-butyrylgalactosamine and N-isobutyrylgalactosamine, preferably N-acetylgalactosamine.

[0243] Preferably, R1 is

[0244] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R2 is selected from hydroxyl, carboxyl, -OR p 、-OPG2、 Preferably selected from -OR p , Preferably selected from Preferably selected from Preferably Preferably

[0245] Preferably, for Preferably

[0246] In a more specific embodiment, the present invention provides a compound of the above formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein PG1 is selected from C 1-6 Alkyl, benzyl, allyl, Among them, R 2s are independently selected from H, halogen, C 1-10 Alkyl and C 1-10 Haloalkyl, q is 0, 1, 2, 3, 4 or 5.

[0247] Preferably, PG1 is selected from methyl, tert-butyl, benzyl, allyl, Preferably selected from Preferably Preferably

[0248] Preferably, R 2s are independently selected from H, halogen, C 1-6 Alkyl and C 1-6 The haloalkyl group is preferably H or halogen, preferably F.

[0249] In a more specific embodiment, the present invention provides a compound of formula (I) as described above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein PG2 is selected from: trimethylsilyl (TMS), triethylsilyl (TES), dimethylisopropylsilyl (DMIPS), diethylisopropylsilyl (DEIPS), tert-butyldimethylsilyl ether (TBDMS), tert-butyldiphenylsilyl (TBDPS), triisopropylsilyl ether (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, dimethoxytrityl (DMT), monomethoxytrityl (MMT), 9-phenylxanthen-9-yl (Pixyl), 9-(p-methoxyphenyl)xanthen-9-yl (Mox), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr), 4,4',4″-trimethoxytrityl (TMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), and p-methoxybenzyloxymethyl (PMBM).

[0250] Preferably, PG2 is selected from dimethoxytrityl (DMT), monomethoxytrityl (MMT), 9-phenylxanthen-9-yl (Pixyl), 9-(p-methoxyphenyl)xanthen-9-yl (Mox), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr), 4,4',4"-trimethoxytrityl (TMTr), tert-butyldimethylsilyl ether (TBMDS), tert-butyldiphenylsilyl (TBDPS) and triisopropylsilyl ether (TIPS), preferably selected from dimethoxytrityl (DMT), monomethoxytrityl (MMT), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr) and 4,4',4"-trimethoxytrityl (TMTr), preferably 4,4'-dimethoxytrityl (DMTr).

[0251] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein PG3 is independently selected from 2-cyanoethyl and C 1-6The alkyl group is preferably selected from 2-cyanoethyl and methyl, and is preferably 2-cyanoethyl.

[0252] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R and R' are each independently C 1-6 The alkyl group is preferably an isopropyl group.

[0253] In a more specific embodiment, the present invention provides a compound of the above formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R, R' and the N atom to which they are attached optionally form a 5-6 membered heterocyclic group together.

[0254] In a more specific embodiment, the present invention provides a compound of the above formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R p for

[0255] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein M 3a Independently selected from chemical bonds, -O-, -S-, -NR6-, -CHR7-, -C(O)-, -S(O) 1-2 -, -NHC(O)-, -C(O)NH-, -NHS(O) 1-2 -、-S(O) 1-2 NH-, -OC(O)-, -C(O)O-, -OS(O) 1-2 -and-S(O) 1-2 O-, preferably selected from a chemical bond, -O-, -S-, -NR6-, -C(O)-, -NHC(O)-, -C(O)NH-, -OC(O)- and -C(O)O-, preferably selected from a chemical bond, -NHC(O)-, -C(O)NH-, -OC(O)- and -C(O)O-, preferably selected from a chemical bond, -NHC(O)- and -C(O)NH-, preferably a chemical bond or -C(O)NH-.

[0256] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein M 3b Independently selected from -O-, -S-, -NR6-, -CHR7-, -CH=CH-, -C≡C-, -C(O)-, -S(O) 1-2 -, -C(O)NH-, -NHC(O)-, -OC(O)-, -C(O)O-, -S(O) 1-2 O-Y1-、-OS(O)1-2 -Y1-, -OS(O) 1-2 O-、-S(O) 1-2 NH-Y1-, -NHS(O) 1-2 -Y1-, phenylene, 5-6 membered heteroarylene,

[0257] Preferably, M 3b independently selected from -O-, -S-, -NR6-, -CHR7-, -CH=CH-, -C≡C-, -C(O)-, -C(O)NH-, -NHC(O)-, -OC(O)-, -C(O)O-, -OS(O)2-Y1-, -OS(O)2O-, -NHS(O)2-Y1-, 5-6 membered heteroarylene, It is preferably selected from -C(O)NH-, -NHC(O)-, -OC(O)-, -C(O)O-, preferably -C(O)NH- or -NHC(O)-, preferably -NHC(O)-.

[0258] Preferably, M 3b The 5-6 membered heteroarylene group is

[0259] Preferably, Y1 is independently phenylene or 5-6 membered heteroarylene, preferably phenylene, preferably

[0260] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: Selected from the following structures: Preferably it is selected from A2, A3, A4, A6 and A9, preferably selected from A2, A3 and A9, preferably A3 or A9.

[0261] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R6 is independently selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-7 Cycloalkyl and 3-7 membered heterocyclic groups, preferably selected from H, C 1-6 Alkyl and C 1-6 Haloalkyl, preferably selected from H, -CH3, More preferably, it is H.

[0262] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R7 is independently selected from H, halogen, CN, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-7 Cycloalkyl, 3-7 membered heterocyclyl, phenyl and 5-6 membered heteroaryl, preferably selected from H, halogen, CN, C 1-6 Alkyl and C 1-6 Haloalkyl, preferably selected from H, halogen, C 1-6 Alkyl and C 1-6 Halogenated alkyl.

[0263] Preferably, one or more methylene groups in the alkyl or haloalkyl in R7 are optionally and independently replaced by a group selected from the following: -C(O)-, -C(O)NH-, -NHC(O)-, -OC(O)-, -O-, -C(O)O-, -S-, -S-(CH2) 0-3 -S-, -NR6-, phenylene, 5-6 membered heteroarylene, -CH=CH-, -C≡C- and -O-(CH2) 1-3 -O- is preferably selected from -C(O)-, -C(O)NH-, -NHC(O)-, -OC(O)-, -O-, -C(O)O-, -S-, -SS-, -NR6-, -CH=CH-, -C≡C- and -O-(CH2)2-O-.

[0264] Preferably, the halogen in R7 is F, Cl or Br.

[0265] In a more specific embodiment, the present invention provides a compound of the above formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein n2 is independently 0, 1, 2, 3, 4, 5 or 6, preferably 0, 1, 2 or 3, preferably 0 or 3.

[0266] In a more specific embodiment, the present invention provides a compound of the above formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein n3 is independently 1, 2 or 3, preferably 2.

[0267] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein R4 is selected from H, C 1-5 Alkyl, C 1-5 Haloalkyl and C 1-5 Alkoxy, preferably selected from H, C 1-3 Alkyl and C 1-3 Halogenated alkyl, preferably H.

[0268] In a more specific embodiment, the present invention provides a compound of the above formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein Link1, Link2 and Link3 are each independently C 1-10 Straight chain alkylene, preferably C 1-6 Straight chain alkylene, preferably C 4-6 The straight chain alkylene group is preferably -(CH2)4- or -(CH2)6-.

[0269] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein Link4 is C 1-20 Straight chain alkylene, preferably C 1-17 Straight chain alkylene, preferably C 1-10 Straight chain alkylene, preferably C 3-20 Straight chain alkylene, preferably C 3-17 Straight chain alkylene, preferably C 3-10 Straight chain alkylene.

[0270] Preferably, one or more methylene groups, preferably 1, 2 or 3 methylene groups, preferably 1 methylene group in the linear alkylene group in Link4 are optionally and independently replaced by R*.

[0271] Preferably, R* is independently selected from: -O-, -NR6-, -CHR7-, -C(O)-, -NR6C(O)-, -C(O)NR6-, -OC(O)- and -C(O)O-, preferably -C(O)NH-, -NHC(O)-, -C(O)O- and -OC(O)-, preferably -C(O)NH- and -C(O)O-, preferably -C(O)NH- and -NHC(O)-, preferably -C(O)NH-.

[0272] Preferably, R* is separated from the carbonyl group on the left side of Link4 by 1-15 methylene groups, preferably 1-12 methylene groups, preferably 1-10 methylene groups, preferably 1-3 methylene groups, preferably 3-10 methylene groups.

[0273] R and R2 are separated by 1-12 methylene groups, preferably 1-10 methylene groups, and preferably 1-6 methylene groups.

[0274] Preferably, Link4 is selected from -(CH2)3-, -(CH2) 10 -, -(CH2)3-C(O)NH-(CH2)6- and -(CH2) 10 -C(O)NH-(CH2)6-, preferably selected from -(CH2)3-, -(CH2) 10- and -(CH2)3-C(O)NH-(CH2)6-, preferably -(CH2)3- or -(CH2) 10 -, preferably selected from -(CH2) 10 -, -(CH2)3-C(O)NH-(CH2)6- and -(CH2) 10 -C(O)NH-(CH2)6-, preferably -(CH2) 10 -or-(CH2) 10 -C(O)NH-(CH2)6-.

[0275] In a more specific embodiment, the present invention provides a compound of the above formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein m1, m2 and m3 are each independently 1, 2 or 3, preferably 1.

[0276] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, which has the following structural formula:

[0277]

[0278] in,

[0279] n2 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0280] The remaining groups are as defined in the present invention.

[0281] In a more specific embodiment, the present invention provides a compound of formula (II) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein:

[0282] R2 is selected from hydroxyl, carboxyl, -OR p 、-OPG2、

[0283] PG1 is a carboxyl protecting group, preferably selected from methyl, tert-butyl, benzyl, allyl,

[0284] R 2s are independently selected from H, halogen, C 1-6 Alkyl and C 1-6 Haloalkyl;

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

[0286] PG2 is a hydroxy protecting group, preferably selected from dimethoxytrityl (DMT), monomethoxytrityl (MMT), 9-phenylxanthen-9-yl (Pixyl), 9-(p-methoxyphenyl)xanthen-9-yl (Mox), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr), 4,4',4"-trimethoxytrityl (TMTr), tert-butyldimethylsilyl ether (TBMDS), tert-butyldiphenylsilyl (TBDPS) and triisopropylsilyl ether (TIPS);

[0287] R p for

[0288] PG3 is independently selected from phosphate protecting groups, preferably selected from 2-cyanoethyl and methyl, more preferably 2-cyanoethyl;

[0289] R and R' are each independently an amino protecting group, preferably C 1-6 Alkyl, preferably isopropyl;

[0290] M 3a is selected from the group consisting of a chemical bond, -NHC(O)-, -C(O)NH-, -OC(O)- and -C(O)O-, preferably selected from the group consisting of a chemical bond, -NHC(O)- and -C(O)NH-;

[0291] n1 is independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0292] n2 is independently 0, 1, 2, 3, 4, 5 or 6;

[0293] n3 is independently 1, 2, 3, 4, 5 or 6;

[0294] m1, m2 and m3 are each independently 1, 2, 3, 4 or 5;

[0295] Link4 is selected from C 1-25 Straight chain alkylene, preferably C 1-20 Straight chain alkylene, wherein 1, 2 or 3 methylene groups in the straight chain alkylene are optionally and independently replaced by R*, and R* is independently selected from: -C(O)NH-, -NHC(O)-, -C(O)O- and -OC(O)-, preferably -C(O)NH- and -NHC(O)-;

[0296] The condition is that the methylene group connected to the carbonyl group on its left side in Link4 is not replaced by R*.

[0297] In a more specific embodiment, the present invention provides a compound of formula (II) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein:

[0298] R2 is selected from -OR p , Preferably selected from Preferably

[0299] PG1 is selected from

[0300] R 2s are independently H or halogen, preferably F;

[0301] q is 0, 1, 2, 3, 4, or 5;

[0302] PG2 is selected from dimethoxytrityl (DMT), monomethoxytrityl (MMT), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr) and 4,4',4"-trimethoxytrityl (TMTr), preferably 4,4'-dimethoxytrityl (DMTr);

[0303] R p for

[0304] PG3 is 2-cyanoethyl;

[0305] R and R' are each independently C 1-4 Alkyl, preferably isopropyl;

[0306] M 3a is a chemical bond or -C(O)NH-;

[0307] n1 is independently 4, 5 or 6;

[0308] n2 is independently 0, 1, 2 or 3;

[0309] n3 is independently 1 or 2;

[0310] m1, m2 and m3 are each independently 1, 2 or 3, preferably 1;

[0311] Link4 is selected from C 1-17 Straight chain alkylene, preferably selected from C 1-10 A straight chain alkylene group, wherein one methylene group in the straight chain alkylene group is optionally replaced by R*, and R* is -C(O)NH-.

[0312] Preferably, R* is separated from the carbonyl group on the left side of Link4 by 1-12 methylene groups, preferably 1-10 methylene groups, preferably 1-3 methylene groups;

[0313] R* is separated from R2 by 1 to 12 methylene groups, preferably 1 to 10 methylene groups, and preferably 1 to 6 methylene groups.

[0314] In a more specific embodiment, the present invention provides a compound of formula (II) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein:

[0315] R2 is selected from Preferably

[0316] M 3a is a chemical bond or -C(O)NH-;

[0317] n1 is independently 4 or 6;

[0318] n2 is independently 0 or 3;

[0319] n3 is 2;

[0320] m1, m2 and m3 are 1;

[0321] Link4 is selected from -(CH2)3-, -(CH2) 10 -、-(CH2)3-C(O)NH-(CH2)6- or -(CH2) 10 -C(O)NH-(CH2)6-, preferably -(CH2)3-, -(CH2) 10 - and -(CH2)3-C(O)NH-(CH2)6-, preferably -(CH2)3- or -(CH2) 10 -.

[0322] In a more specific embodiment, the present invention provides a compound of formula (I) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the compound is selected from the following:

[0323]

[0324]

[0325] In a more specific embodiment, the present invention provides a conjugate having a structure shown in formula (III):

[0326]

[0327] in,

[0328] L is a structure represented by formula (I') or formula (II'):

[0329]

[0330] Selected from Preferably selected from Preferably selected from Preferably selected from Preferably selected from

[0331] R D is a drug molecule independently selected from a cytotoxic agent, a chemotherapeutic agent, a somatostatin, a toxin, a radioisotope, and an oligonucleotide chain;

[0332] d is 1 or 2, preferably 1;

[0333] The remaining groups are as defined herein.

[0334] Preferably, Link4 is selected from C 5-25 Straight chain alkylene, preferably C 5-20 Straight chain alkylene, preferably C 8-20 Straight chain alkylene, preferably C 10-17 Straight chain alkylene.

[0335] Preferably, one or more methylene groups, preferably 1, 2 or 3 methylene groups, preferably 1 methylene group in Link4 are optionally and independently replaced by R*, and R* is independently selected from: -C(O)NH-, -NHC(O)-, -C(O)O- and -OC(O)-, preferably -C(O)NH- and -C(O)O-, preferably -C(O)NH- and -NHC(O)-, preferably -C(O)NH-.

[0336] Preferably, R* is separated from the carbonyl group on the left side of Link4 by 1-15 methylene groups, preferably by 1-12 methylene groups, preferably by 1-10 methylene groups, preferably by 3-10 methylene groups;

[0337] R* is separated from R2 by 1 to 12 methylene groups, preferably 1 to 10 methylene groups, and preferably 1 to 6 methylene groups.

[0338] Preferably, Link4 is selected from -(CH2) 10 -, -(CH2)3-C(O)NH-(CH2)6- and -(CH2) 10 -C(O)NH-(CH2)6-, preferably -(CH2) 10 -or-(CH2) 10 -C(O)NH-(CH2)6-.

[0339] In a more specific embodiment, the present invention provides a conjugate of the above formula (III), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein RD It is an oligonucleotide chain, optionally, the oligonucleotide chain is a single-stranded oligonucleotide or a double-stranded oligonucleotide or a combination thereof.

[0340] Preferably, M2 is linked to the 5' end, 3' end or any nucleotide between the 5' end and the 3' end of at least one chain of the oligonucleotide chain through a phosphate bond, preferably linked to the 5' end or 3' end of at least one chain, preferably linked to the 5' end of at least one chain, and preferably linked to the 3' end of at least one chain.

[0341] Preferably, M2 is linked to the sense strand of the oligonucleotide chain via a phosphate bond.

[0342] Preferably, the phosphate bond is a phosphodiester bond or a modified phosphate bond, preferably a phosphodiester bond; preferably, the modified phosphate bond is selected from a thio-modified phosphate bond and an amino-modified phosphate bond, preferably a thio-modified phosphate bond.

[0343] Preferably, the Selected from the following structures:

[0344]

[0345] Preferably selected from

[0346] Preferably selected from

[0347] in, represents an oligonucleotide chain.

[0348] In a more specific embodiment, the present invention provides the conjugate of formula (III) above, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the oligonucleotide chain comprises unmodified nucleotides and / or modified nucleotides.

[0349] Preferably, the modified nucleotides are each independently selected from 2′-methoxyethyl modified nucleotides, 2′-O-alkyl modified nucleotides, 2′-O-allyl modified nucleotides, 2′-C-allyl modified nucleotides, 2′-fluoro modified nucleotides, 2′-deoxy modified nucleotides, 2′-hydroxy modified nucleotides, thiophosphate backbone modified nucleotides, locked nucleotide modified nucleotides, glycerol nucleic acid (glycol nucleic acid, GNA) modified nucleotides and unlocked nucleic acid (UNA) modified nucleotides, preferably selected from 2′-O-alkyl modified nucleotides, 2′-fluoro modified nucleotides, 2′-deoxy modified nucleotides and thiophosphate backbone modified nucleotides.

[0350] Preferably, the 2′-O-alkyl group is 2′-OC 1-6 The alkyl group is preferably a 2'-O-methyl group.

[0351] Preferably, the length of the oligonucleotide chain is 5-100 bp.

[0352] In a more specific embodiment, the present invention provides the above-mentioned conjugate of formula (III), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the oligonucleotide chain is siRNA, ASO or microRNA, preferably siRNA.

[0353] In a more specific embodiment, the present invention provides the above-mentioned conjugate of formula (III), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the oligonucleotide chain has a terminal modification, and the terminal modification is selected from cholesterol, polyethylene glycol, a fluorescent probe, biotin, a polypeptide, a vitamin, a tissue targeting molecule, or a combination thereof.

[0354] In a more specific embodiment, the present invention provides a conjugate of the above-mentioned formula (III), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the ribose-phosphate backbone of the oligonucleotide chain is replaced by a polypeptide nucleic acid (PNA) or a morpholino antisense nucleotide (PMO).

[0355] In a more specific embodiment, the present invention provides a conjugate of the above formula (III), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein L is selected from the following structures:

[0356]

[0357]

[0358] In a more specific embodiment, the present invention provides a conjugate of the above formula (III), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the conjugate is selected from the following:

[0359]

[0360]

[0361] In a more specific embodiment, the present invention also provides a pharmaceutical composition comprising the conjugate of the present invention, and optionally a pharmaceutically acceptable carrier, excipient, adjuvant or vehicle, and optionally other therapeutic agents.

[0362] In a more specific embodiment, the present invention also provides use of the conjugate of the present invention or the pharmaceutical composition of the present invention in the preparation of a medicament for treating or preventing a disease associated with the expression or overexpression of a gene in hepatocytes.

[0363] In a more specific embodiment, the present invention also provides a method for treating or preventing a disease associated with the expression or overexpression of a gene in hepatocytes in a subject, comprising administering the conjugate of the present invention or the pharmaceutical composition of the present invention to the subject.

[0364] In a more specific embodiment, the present invention also provides the conjugate of the present invention or the pharmaceutical composition of the present invention for use in treating or preventing diseases associated with the expression or overexpression of a gene in hepatocytes.

[0365] In a more specific embodiment, the gene is selected from the HBV genome, HCV genome, PCSK9, xanthine oxidase, URAT1, APOB, liver fibrosis-related genes (AP3S2, AQP2, AZINI, DEGSI, STXBP5L, TLR4, TRPM5), non-alcoholic fatty liver disease-related genes (PNPLA3, FDFTl), primary biliary cirrhosis-related genes (HLA-DQB1, IL-12, IL-12RB2), or a combination thereof.

[0366] In a more specific embodiment, the disease is selected from: hereditary angioedema, familial tyrosinemia type I, Alagille syndrome, alpha-1-antitrypsin deficiency, bile acid synthesis and metabolism defects, biliary atresia, cystic fibrosis liver disease, idiopathic neonatal hepatitis, mitochondrial liver disease, progressive familial intrahepatic cholestasis, primary sclerosing cholangitis, transthyretin amyloidosis, hemophilia, homozygous familial hypercholesterolemia, hyperlipidemia, hepatitis B (HBV), hepatitis C (HCV), steatohepatitis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), hyperglycemia or diseases involving abnormally increased hepatic glucose production similar to type III diabetes, hepatitis and hepatic porphyrin.

[0367] In a more specific embodiment, the present invention also provides a compound of formula (C):

[0368]

[0369] Among them, PG2 and PG4 are protecting groups.

[0370] Preferably, PG2 is a hydroxyl protecting group and PG4 is an amino protecting group;

[0371] Preferably, the PG2 is selected from trimethylsilyl (TMS), triethylsilyl (TES), dimethylisopropylsilyl (DMIPS), diethylisopropylsilyl (DEIPS), tert-butyldimethylsilyl ether (TBDMS), tert-butyldiphenylsilyl (TBDPS) and triisopropylsilyl ether (TIPS), preferably tert-butyldimethylsilyl ether (TBDMS);

[0372] Preferably, the PG4 is selected from benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), 9-fluorenylmethoxycarbonyl (Fmoc), allyloxycarbonyl (Alloc), 2-(trimethylsilyl)ethoxycarbonyl (Teoc), methoxycarbonyl and ethoxycarbonyl, preferably tert-butyloxycarbonyl (Boc).

[0373] In a more specific embodiment, the present invention also provides a method for preparing a compound of formula (C), which comprises the following steps: reacting compound A with compound B to generate compound C:

[0374]

[0375] Wherein, PG2 and PG4 are as defined in the present invention.

[0376] Preferably, the reaction conditions of the method include a condensing agent and a base.

[0377] Preferably, the condensing agent is selected from carbodiimide condensing agents, onium salt condensing agents and organic phosphorus condensing agents.

[0378] Preferably, the reaction conditions further include a condensation activator.

[0379] Preferably, the reaction conditions further include a solvent.

[0380] Preferably, the method is carried out in a nitrogen atmosphere.

[0381] Preferably, the base is an organic amine base, preferably DIEA (N,N-diisopropylethylamine), TEA (triethylamine), NMM (N-methylmorpholine), DBU (1,8-diazabicyclo[5.4.0]undec-7-ene), DMAP (4-dimethylaminopyridine), pyridine, 2,6-lutidine and 2,2,6,6-tetramethylguanidine, preferably DIEA.

[0382] Preferably, the carbodiimide condensing agent is selected from DCC (N,N'-dicyclohexylcarbodiimide), DIC (N,N'-diisopropylcarbodiimide) and EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), preferably EDCI.

[0383] Preferably, the onium salt condensing agent is selected from HATU (N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1-yl) uronium hexafluorophosphate), HBTU (benzotriazole-N,N,N′,N′-tetramethyluronium hexafluorophosphate), HCTU (6-chlorobenzotriazole-1,1,3,3-tetramethyluronium hexafluorophosphate), TBTU (2-(1H-benzotriazol L-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate), BOP (Carter's condensing agent) and PyBOP (1H-benzotriazol-1-yloxytripyrrolidino hexafluorophosphate).

[0384] Preferably, the organophosphorus condensing agent is selected from DPP-Cl (diphenylphosphinyl chloride), DPPA (diphenylphosphoryl azide) and BOP-Cl (bis(2-oxo-3-oxazolidinyl)phosphinyl chloride).

[0385] Preferably, the condensation activator is selected from DMAP (4-dimethylaminopyridine), HOBt (1-hydroxybenzotriazole) and HOAt (N-hydroxy-7-azabenzotriazole), preferably HOBt.

[0386] Preferably, the solvent is selected from DCM (dichloromethane), DCE (1,2-dichloroethane), DMF (N,N-dimethylformamide), DMAC (N,N-dimethylacetamide), tetrahydrofuran and 2-methyltetrahydrofuran, preferably DCM.

[0387] Preferably, the molar ratio of compound A:compound B is (0.5-3):1, preferably (1-2):1, preferably (1-1.5):1, preferably 1.1:1.

[0388] Preferably, the molar ratio of condensing agent:compound B is (1-3):1, preferably (1-2):1, preferably (1-1.5):1, preferably 1.2:1.

[0389] Preferably, the molar ratio of condensation activator:compound B is (1-3):1, preferably (1-2):1, preferably (1.2-1.8):1, preferably 1.5:1.

[0390] Preferably, the molar ratio of base:compound B is (0.5-5):1, preferably (1-4):1, preferably (1-3):1, preferably 2:1.

[0391] Preferably, the process is carried out at a temperature of about -20°C to about 50°C, preferably about -10°C to about 25°C, preferably about -10°C to about 0°C.

[0392] In a more specific embodiment, the present invention also provides a compound of formula (D):

[0393]

[0394] In a more specific embodiment, the present invention also provides a method for preparing a compound of formula (D), which comprises the following steps: removing the protecting group of the compound C of the present invention to obtain a compound D:

[0395]

[0396] Wherein, PG2 and PG4 are as defined in the present invention.

[0397] Preferably, the method is carried out under acidic conditions.

[0398] Preferably, the reaction conditions of the method include an acid and a solvent.

[0399] Preferably, the acid is HCl and / or H2SO4, preferably HCl.

[0400] Preferably, the solvent is an organic solvent, preferably an alcohol solvent, preferably selected from methanol, ethanol, propanol, butanol, isopropanol, cyclohexanol, 2-methyl-2-propanol and 2-ethyl-1-propanol, preferably methanol.

[0401] Preferably, the concentration of HCl in the reaction system is 0.1 mol / L-3 mol / L, preferably 0.2 mol / L-2 mol / L, preferably 0.5 mol / L-1.5 mol / L, preferably 0.5 mol / L-1.0 mol / L, preferably 0.75 mol / L.

[0402] Preferably, the method is carried out in a nitrogen atmosphere.

[0403] Preferably, the process is carried out at a temperature of about -20°C to about 50°C, preferably about -10°C to about 25°C, preferably about -10°C to about 0°C.

[0404] The compounds of the invention or the conjugates of the invention may include one or more asymmetric centers and may therefore exist in a variety of stereoisomeric forms, for example, enantiomers and / or diastereoisomeric forms. For example, the compounds of the invention may be individual enantiomers, diastereomers or geometric isomers (e.g., cis and trans isomers), or may be in the form of a mixture 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.

[0405] The compound or conjugate of the present invention may exist in the form of tautomers. Tautomers are functional group isomers produced by the rapid movement of an atom in a molecule between two positions. Tautomers are a special type of functional group isomers. A pair of tautomers can be converted into each other, but usually a relatively stable isomer is its main existing form.

[0406] Pharmaceutical compositions, preparations and kits

[0407] The present invention provides a pharmaceutical composition comprising a compound or conjugate of the present invention (also referred to as an "active ingredient") and a pharmaceutically acceptable carrier or excipient. In some embodiments, the pharmaceutical composition comprises an effective amount of the active ingredient. In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of the active ingredient. In some embodiments, the pharmaceutical composition comprises a preventively effective amount of the active ingredient.

[0408] The pharmaceutically acceptable excipient used in the present invention refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound or conjugate formulated together. The pharmaceutically acceptable carrier, adjuvant or vehicle that can be used in the composition of the present invention includes, but is not limited to, ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffer substances (such as phosphates), glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, silica gel, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and lanolin.

[0409] The present invention also includes a kit (e.g., a pharmaceutical package). The kit provided may include a compound of the invention, other therapeutic agents, and a first and second container (e.g., a vial, an ampoule, a bottle, a syringe, and / or a dispersible package or other suitable container) containing the compound of the invention and other therapeutic agents. In some embodiments, the kit provided may also optionally include a third container containing a pharmaceutical excipient for diluting or suspending the compound of the invention and / or other therapeutic agents. In some embodiments, the compound of the invention and other therapeutic agents provided in the first container and the second container are combined to form a unit dosage form.

[0410] Pharmaceutical composition provided by the invention can be administered by many ways, including but not limited to: oral administration, parenteral administration, inhalation administration, topical administration, rectal administration, nasal administration, oral administration, vaginal administration, administration by implant or other modes of administration. For example, parenteral administration used herein includes subcutaneous administration, intradermal administration, intravenous administration, intramuscular administration, intraarticular administration, intraarterial administration, intrasynovial cavity administration, intrasternal administration, intrathecal administration, intralesional administration and intracranial injection or infusion technology.

[0411] Generally, an effective amount of the compounds provided herein is administered. The amount of the compound actually administered can be determined by a physician according to the relevant circumstances, including the condition being treated, the route of administration selected, the compound actually administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, and the like.

[0412] When used to prevent the conditions described herein, the compounds provided herein are administered to a subject at risk of developing the condition, typically based on the advice of a physician and under the supervision of a physician, at dosage levels as described above. Subjects at risk of developing a specific condition typically include those with a family history of the condition, or those identified by genetic testing or screening as being particularly susceptible to developing the condition.

[0413] The pharmaceutical compositions provided herein can also be administered over a long period of time ("chronic administration"). Chronic administration refers to administration of a compound or a pharmaceutical composition thereof over a long period of time, e.g., 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, etc., or administration may continue indefinitely, e.g., for the rest of the subject's life. In some embodiments, chronic administration is intended to provide a constant level of the compound in the blood over a long period of time, e.g., within a therapeutic window.

[0414] Various methods of administration can be used to further deliver the pharmaceutical composition of the present invention. For example, in some embodiments, the pharmaceutical composition can be administered by push injection, for example, in order to quickly increase the concentration of the compound in the blood to an effective level. The push dose depends on the target systemic level of the active component, for example, the intramuscular or subcutaneous push dose slowly releases the active component, and the push delivered directly to the vein (for example, by IV intravenous drip) can be delivered more quickly, so that the concentration of the active component in the blood is quickly increased to an effective level. In other embodiments, the pharmaceutical composition can be given in a continuous infusion form, for example, by IV intravenous drip, so as to provide a steady-state concentration of the active component in the subject's body. In addition, in other embodiments, the pharmaceutical composition of the push dose can be first given, and then continuous infusion.

[0415] In order to provide blood levels similar to those using an injectable dose, or lower blood levels than using an injectable dose, a transdermal dose is generally selected in an amount of about 0.01 to about 20% by weight, preferably about 0.1 to about 20% by weight, preferably about 0.1 to about 10% by weight, and more preferably about 0.5 to about 15% by weight.

[0416] From about 1 to about 120 hours, especially 24 to 96 hours, the injected dosage level is in the range of about 0.1 mg / kg / hour to at least 10 mg / kg / hour. In order to obtain sufficient steady state levels, a preload bolus of about 0.1 mg / kg to about 10 mg / kg or more may also be given. For a 40 to 80 kg human patient, the maximum total dose cannot exceed about 2 g / day.

[0417] Injectable compositions are typically based on injectable sterile saline or phosphate buffered saline, or other injectable excipients known in the art. As previously mentioned, in such compositions, the active compound is typically a minor component, often about 0.05 to 10% by weight, and the remainder is an injectable excipient, etc.

[0418] The above components of the compositions for injection or topical administration are representative only. Other materials and processing techniques are described in Section 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, Mack Publishing Company, Easton, Pennsylvania, which is incorporated herein by reference.

[0419] The compounds of the invention may also be administered in sustained release form or from a sustained release delivery system. Descriptions of representative sustained release materials can be found in Remington's Pharmaceutical Sciences.

[0420] The present invention also relates to pharmaceutically acceptable formulations of the compounds of the present invention. In one embodiment, the formulation comprises water.

[0421] Drug combinations

[0422] The compounds of the invention or conjugates described herein may be used in combination with one or more other active ingredients in pharmaceutical compositions or methods to treat the diseases and disorders described herein. Other additional active ingredients include other therapeutic agents or agents that mitigate the adverse effects of the intended disease target. The combination may be used to increase efficacy, improve other disease symptoms, reduce one or more side effects, or reduce the required dose of the compounds of the invention. Additional active ingredients may be formulated into pharmaceutical compositions separate from the compounds of the invention or may be included in a single pharmaceutical composition with the compounds of the invention. Additional active ingredients may be administered simultaneously with, before, or after the administration of the compounds of the invention.

[0423] Combination agents include those active ingredients that are known or observed to be effective in treating the diseases and conditions described herein, including those that are effective against another target associated with the disease. For example, the compositions and preparations of the present invention, and the methods of treatment may further include other drugs, such as other agents that can be used to treat or alleviate the target disease or related symptoms or conditions. The pharmaceutical compositions of the present invention may additionally include one or more of the active agents, and the methods of treatment may additionally include administering an effective amount of one or more of the active agents.

[0424] Beneficial effects of the present invention:

[0425] (1) The present invention uses serine as a starting material and generates a new molecular peptide scaffold skeleton with the α carbon atom in serine as the central branch point through a condensation reaction with serinol. The new molecular skeleton satisfies the following structural characteristics: (i) a central branch structure; (ii) a spacer based on multiple peptide bond linkers, which combines flexibility and amphiphilicity; (iii) the spacing distance between the spacer connected between the central branch structure and the GalNAc group is (iv) The length of the spacer can be easily varied.

[0426] (2) The novel three-cluster galactose derivative of the present invention has better hydrophilicity and low toxicity; the formed conjugate has a high affinity for binding to the ASGPR receptor, with a Kd value of less than 10 and can be as low as about 2; compared with the negative control siRNA without GalNAc, the distribution of the conjugate of the present invention in the mouse liver is significantly increased, and it can effectively achieve liver-targeted delivery in vivo and significantly knock down the expression of the target gene mRNA.

[0427] (3) The trivalent GalNAc cluster derivatives based on the novel skeleton of the present invention have a simple synthesis method and are covalently coupled with ASO using a mature phosphoramidite solid phase synthesis coupling strategy. This can improve the efficacy of ASO drugs in clinical practice without increasing the complexity or cost of existing oligonucleotide drug production schemes. Existing, cheap and readily available raw materials can be used for effective assembly synthesis and large-scale production. BRIEF DESCRIPTION OF THE DRAWINGS

[0428] Figure 1 This is the binding curve of G1-12 of the present invention and ASPGR receptor.

[0429] Figure 2 This is the binding curve of G4-12 of the present invention and ASPGR receptor.

[0430] Figure 3 This is the binding curve of G5-12 of the present invention and ASPGR receptor.

[0431] Figure 4 This is the test result of G1-12 of the present invention applied to liver targeting in mice.

[0432] Example

[0433] The technical solution of the present invention is described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0434] Example 1: Synthesis of G1

[0435]

[0436] Compound 1 (100 g) was dissolved in dry DCM (500 mL), and DMAP (1 g) and Et3N (5 eq) were added under nitrogen protection in an ice bath, and TBDMSCl (2.5 eq) was slowly added. After the reaction was complete, the mixture was washed with NaHCO3 solution and chromatographed to obtain compound 2.

[0437] Compound 3 (100 g) was dissolved in anhydrous DMF (500 mL), imidazole (3.2 eq) was added under nitrogen protection in an ice bath, and TBDMSCl (1.4 eq) was slowly added. After the reaction was complete, the mixture was washed with saturated NaHCO3 and saturated brine in sequence, concentrated, and chromatographed to obtain compound 4.

[0438] Compound 4 (50 g) was dissolved in anhydrous DCM (30 mL), HOBt (1.5 eq) was added under the protection of nitrogen in an ice bath, and EDCI (1.2 eq) was slowly added. After reacting for 20 min, compound 2 (1.1 eq) was added under the protection of nitrogen in an ice bath, and DIEA (2 eq) was added. After the reaction was complete, the mixture was washed with saturated NaHCO3, 10% aqueous citric acid solution, and saturated brine in sequence, concentrated, and chromatographed to obtain compound 5.

[0439] Compound 5 (30 g) was dissolved in MeOH (60 mL), and a MeOH solution of HCl (20 mL, 3 mol / L) was added under nitrogen protection in an ice bath. The mixture was reacted at room temperature. After the reaction was complete, the mixture was concentrated and washed with ethyl acetate to obtain compound 6.

[0440] Compound 6 (20 g) was dissolved in anhydrous DMSO (100 mL), NaOH (1.3 eq) was added under nitrogen protection in an ice bath, and tert-butyl acrylate (6 eq) was slowly added. After the reaction was complete, the mixture was washed with EA and saturated brine in sequence, concentrated, and chromatographed to obtain compound 7.

[0441] Compound 7 (15 g) was dissolved in anhydrous DCM (300 mL), and monomethyl dodecanoate (1.1 eq) and HOBt (1.5 eq) were added under nitrogen protection in an ice bath, and EDCI (1.2 eq) was slowly added. After reacting for 20 min, DIEA (2 eq) was added, and the mixture was washed with saturated NaHCO3, 10% aqueous citric acid solution, and saturated brine in sequence, concentrated, and chromatographed to obtain compound 8.

[0442] Compound 8 (15 g) was dissolved in HCOOH (30 mL). After the reaction was complete, the mixture was concentrated and chromatographed to obtain compound 9.

[0443] Compound 9 (10 g) was dissolved in dry DMF (60 mL), and HOBt (1.25 eq), EDCI (1.2 eq) and N-tert-butyloxycarbonyl-1,3-propanediamine (1.2 eq) were added. DIEA (2 eq) was added under nitrogen protection in an ice bath. After the reaction was complete, the mixture was washed with saturated sodium bicarbonate solution and 0.5 N hydrochloric acid solution in sequence, and compound 10 was obtained by chromatography.

[0444] Compound 10 (9.3 g) was dissolved in dry DCM (60 mL), and a solution of HCl in MeOH (20 mL, 2 mol / L) was added under nitrogen protection in an ice bath. After the reaction was completed, compound 11 was obtained.

[0445] N-Acetylgalactosamine (15 g) was dissolved in anhydrous DMF (100 mL), HOBt (1.5 eq) was added under nitrogen protection in an ice bath, EDCI (1.2 eq) was slowly added, and the mixture was reacted for 20 min. DIEA (2 eq) and compound 11 (9.3 g) were added for reaction. After the reaction was complete, the mixture was washed with saturated NaHCO3, 0.5 N aqueous hydrochloric acid solution, and saturated brine in sequence, and then concentrated and chromatographed to obtain compound 12.

[0446] Compound 12 (4 g) was dissolved in dry MeOH (20 mL), and NaOMe (30%, MeOH, 3 mL) was added under nitrogen protection in an ice bath. After the reaction was complete, the crude product was concentrated to obtain a crude product. The crude product was added to MeOH / H2O and reacted at room temperature. After the reaction was complete, the pH was adjusted to neutral with HCl salt of Et3N, and the crude product was concentrated to obtain a crude product. The crude product was then added to Py (20 mL) and Ac2O (5 mL) and reacted at room temperature. After the reaction was complete, compound 13 was obtained by chromatography.

[0447] Compound 13 (1 g) was dissolved in DCM (10 mL), and EDCI (1.5 eq) and pentafluorophenol (1.5 eq) were added. After the reaction was completed, the mixture was washed with saturated sodium bicarbonate solution, concentrated, and chromatographed to obtain compound G1 (0.5 g) with a yield of 50%.

[0448] 1H NMR (500MHz, CDCl3) δ: 7.61–7.30 (m, 3H), 5.31–5.19 (m, 3H), 5.03 (d, J = 11.1Hz ,3H),4.49(d,J=8.3Hz,3H),4.15–3.96(m,9H),3.86(m,6H),3.71–3.55(m,9H), 3.50-3.39(m,12H),3.16(m,6H),2.59(t,J=7.6Hz,2H),2.35(dt,J=4.9Hz,6H), 2.26–2.03(m,20H),2.03–1.81(m,30H),1.55(m,8H),1.19(m,20H), ESI-Tof-MS m / z:2264.10[M+Cl] - ; C 99 H 150 F5N 11 O 40 .

[0449] Example 2: Synthesis of G2

[0450]

[0451] Compound G5-11 (2 g) was dissolved in DCM (10 mL), and HBTU (1.2 eq), HOBT (3 eq), DIPEA (3 eq), and proline (1.05 eq) were added. After the reaction was completed at room temperature, the mixture was washed with saturated sodium bicarbonate solution and concentrated to obtain a crude product, which was then chromatographed to obtain compound G2-12.

[0452] Compound G2-12 (1 g) was dissolved in anhydrous DCM (10 mL), DIPEA (3 eq) was added, and phosphoramidite monomer (1.5 eq) was added under nitrogen protection at low temperature. After the reaction was completed at room temperature, it was washed with saturated sodium bicarbonate solution, concentrated and chromatographed to obtain G2 (0.5 g) with a yield of 41%.

[0453] 1H NMR (500MHz, CDCl3): δ7.29(t,J=7.9Hz,2H),7.24–7.06(m,10H),6.91(s,1H),6.78–6.69(m,6H),6.62 –6.41(m,3H),5.28(s,3H),5.19(d,J=9.2Hz,3H),4.61(s,3H),4.45(m,2H),4.07(m,6H),3.93(m,3H), 3.89–3.76(m,6H),3.69(m,12H),3.57–3.28(m,10H),3.13(m,6H),3.04(m,2H),2.52(dd,J=12.0,5.7H z,2H),2.37(m,6H),2.19(m,4H),1.99(m,36H),1.72(m,16H),1.60–1.36(m,16H),1.34–0.93(m,22H); 31 PNMR (202MHz, CDCl3) δ: 147.37, (s). ESI-Tof-MS m / z: 2491.25[MH] - ; C 122 H 186 N 11 O 41 P.

[0454] Example 3: Synthesis of G3

[0455]

[0456] Compound G4-11 (2 g) was dissolved in DCM (10 mL), and EDCI (1.5 eq) was added, followed by HOBT (2 eq), DIPEA (3 eq) and compound b (1.2 eq). After the reaction was completed at room temperature, the mixture was washed with saturated sodium bicarbonate solution, concentrated and chromatographed to obtain G3-12 (1.5 g).

[0457] Compound G3-12 (2 g) was dissolved in THF (20 mL), and triethylamine trihydrofluoride (1.5 eq) was added. After the reaction was completed at room temperature, it was concentrated and DCM was added. The mixture was washed with saturated sodium bicarbonate solution, concentrated, and chromatographed to obtain G3-13 (1.3 g).

[0458] Compound G3-13 (1.2 g) was dissolved in anhydrous DCM (10 mL), DIPEA (3 eq) was added, and phosphoramidite monomer (1.5 eq) was added under nitrogen protection at low temperature. After the reaction was completed at room temperature, it was washed with saturated sodium bicarbonate solution, concentrated and chromatographed to obtain compound G3 (0.6 g) with a yield of 46%.

[0459] 1H NMR (500MHz, CDCl3): δ7.16 (s, 1H), 7.04 (t, J = 14.7Hz, 2H), 6.83 (d, J = 5.5Hz, 2H), 6.69 (d, J = 5.7Hz, 1H), 6.56 (d, J = 8.8 Hz,2H),6.42(s,1H),5.35(d,J=2.5Hz,3H),5.30–5.23(m,3H),4.68(d,J=8.3Hz,3H),4.21–4.06(m,8H),3.99(dd,J=19. 4,8.8Hz,3H),3.94–3.83(m,6H),3.63(m,6H),3.46(dd,J=15.8,6.6Hz,4H),3.34–3.16(m,16H),2.68(t,J=7.3Hz,2H),2 .42(t,J=5.2Hz,6H),2.33–2.22(m,4H),2.21–1.88(m,38H),1.79(m,14H),1.63–1.43(m,18H),1.30(d,J=43.5Hz,12H); 31 P NMR (202MHz, CDCl3) δ: 147.37, (s). ESI-Tof-MS m / z: 2033.40[MH] - ; C 93 H 156 N 11 O 36 P.

[0460] Example 4: Synthesis of G4

[0461]

[0462] Compound 7 (25 g) was dissolved in anhydrous DCM (50 mL), and Na2CO3 (200 mL, 25% aqueous solution) was added under nitrogen protection in an ice bath, and CbzCl (2 eq) was slowly added. After the reaction was complete, the mixture was washed with saturated brine, concentrated, and chromatographed to obtain G4-8 (30 g).

[0463] Compound G4-8 (8 g) was dissolved in HCOOH (30 mL). After the reaction was complete, the mixture was concentrated and chromatographed to obtain G4-9 (5 g).

[0464] Compound G4-9 (8 g) was dissolved in anhydrous DCM (50 mL), HOBt (3.3 eq) was added under nitrogen protection in an ice bath, EDCI (3.3 eq) was slowly added, and the reaction was continued for 20 min. N-acetylgalactosamine (3.3 eq) and DIEA (6 eq) were added. After the reaction was complete, the mixture was washed with saturated NaHCO3, 0.5 mol / L hydrochloric acid solution, and saturated brine in sequence, and concentrated and chromatographed to obtain G4-10 (6.4 g).

[0465] Compound G4-10 (3 g) was dissolved in anhydrous THF (50 mL), succinic anhydride (1.5 eq) was added, and 10% Pd / C (1 g) was slowly added. After the reaction was completed in a hydrogen environment, G4-11 (2.5 g) was obtained by filtration, concentration and chromatography.

[0466] Compound G4-11 (1 g) was dissolved in DCM (10 mL), and EDCI (1.5 eq) and pentafluorophenol (1.5 eq) were added. After the reaction was completed, the mixture was washed with a saturated sodium bicarbonate solution, concentrated, and chromatographed to obtain G4 (0.5 g) with a yield of 40%.

[0467] 1 H NMR (500MHz, CDCl3): δ7.29–7.20(m,2H),7.11(m,1H),6.91–6.68(m,3H),6.56–6.39(m,2H),5.34(d,J=2.5Hz,3H ),5.25(d,J=11.1Hz,3H),4.66(d,J=8.3Hz,3H),4.71–4.50(m,2H),4.13(tt,J=18.0,8.9Hz,6H),3.98(dd,J=18. 4,9.2Hz,3H),3.88(ddd,J=15.5,11.8,6.3Hz,6H),3.79–3.63(m,6H),3.56–3.35(m,9H),3.21(td,J=12.5,6.2Hz ,6H),2.76(t,J=7.3Hz,2H),2.43(dd,J=15.3,7.7Hz,8H),2.19–1.86(m,38H),1.62–1.42(m,12H),1.28(m,12H).

[0468] ESI-MS m / z:1957.30[MH]-;C 86 H 127 F5N8 37 .

[0469] Example 5: Synthesis of G5

[0470]

[0471] Compound 9 (8 g) was dissolved in anhydrous DCM (50 ml), HOBt (1.5 eq) was added under nitrogen protection in an ice bath, EDCI (1.2 eq) was slowly added, and the reaction was continued for 20 min. N-acetylgalactosamine (3.3 eq) and DIEA (2 eq) were added. After the reaction was completed, the mixture was washed with saturated NaHCO3, 0.5 mol / L hydrochloric acid solution, and saturated brine in sequence, and concentrated to obtain a crude product, which was then chromatographed to obtain G5-10.

[0472] Compound G5-10 (4 g) was dissolved in dry MeOH (20 mL), and NaOMe (30% in MeOH, 3 ml) was added under the protection of nitrogen in an ice bath. After the reaction was complete, the mixture was concentrated to obtain a crude product, which was added to a mixed solution of MeOH / H2O = 1:1, and reacted at room temperature for 16 hours. The pH was adjusted to neutral with Et3N HCl salt, and the crude product was concentrated. Py (20 ml) and Ac2O (5 ml) were added and reacted at room temperature for 24 hours, and G5-11 (2 g) was obtained by concentration and chromatography.

[0473] Compound G5-11 (1 g) was dissolved in DCM (10 mL), and EDCI (1.5 eq) and pentafluorophenol (1.5 eq) were added. After the reaction was completed, the product was washed with saturated sodium bicarbonate solution, concentrated, and purified by chromatography to obtain G5 (0.5 g) with a yield of 40%.

[0474] 1 H NMR (500MHz, CDCl3): δ7.25–7.17(m,1H),6.93–6.73(m,4H),6.54–6.42(m,3H),5.34(d,J=2 .5Hz,3H),5.29–5.21(m,3H),4.66(d,J=8.2Hz,3H),4.20–4.06(m,8H),4.03–3.80(m,6H),3 .80–3.63(m,8H),3.58–3.34(m,10H),3.31–3.11(m,6H),2.64(t,J=7.4Hz,2H),2.52–2.33( m,6H),2.31–2.20(m,8H),2.09–1.88(m,27H),1.68–1.43(m,16H),1.34(m,24H).ESI-Tof-MS m / z:2057.49[M+H] + ,2079.48[M+Na] + ; C 93 H 141 F5N8 37 .

[0475] Biological experiments

[0476] Experimental Example 1 Affinity determination of hepatocyte ASGPR receptor

[0477] Referring to Chinese invention patent CN109957567B, the synthetic siRNA sequence is as follows:

[0478] dG*fU*dAfUdGfUfUdGfCfCfCdGfUfUfUdGfU*fC*fC(SEQ ID NO.1)

[0479] mG*fG*mA*fCmAfAmAfCmGfGmGfCmAfAmCfAmUfA*mC*mC*mU(SEQ ID NO.2)

[0480] The compounds G1, G4, G5 obtained in Examples 1, 4, and 5 were respectively connected to the above siRNA to form conjugates, as shown in Table 1.

[0481] Table 1 Conjugate structures

[0482]

[0483] Note: A is adenine ribonucleotide, G is guanine ribonucleotide, C is cytosine ribonucleotide, U is uracil ribonucleotide, d is deoxyribonucleotide modification, f is 2'-fluoro modification, m is 2'-O-methyl modification, * is thiophosphorylation backbone modification, and Cy5 is fluorescent label Cy5.

[0484] Among them, the structure of G1-12 is shown in Z1. When G4 and G5 are connected to siRNA, the phosphate bond needs to be modified. In this embodiment, the amino group of C6 is used. Modification, so the structure of G4-12 is shown as Z4, and the structure of G5-12 is shown as Z5.

[0485] 1. Dilute GalNac-siRNA

[0486] G1-12, G4-12, G5-12 (1uM) were diluted in the following gradients:

[0487] Table 2

[0488]

[0489]

[0490] 2. Cell Preparation

[0491] Dilute the cells to a concentration of 4*10 using DMEM medium containing 2% FBS. 5 Pieces / mL.

[0492] 3. Mixing and incubation

[0493] For each sample, 200uL cells were mixed with 200uL siRNA, and for the blank control group, 200uL cells were mixed with 200uL culture medium. Added to 3 wells of a U-bottom 96-well plate, 100uL per well. After adding all siRNA samples of various concentrations, placed them in an ice box, placed on a decolorizing shaker, and incubated for 2 hours away from light.

[0494] 4. Detection

[0495] Centrifuge the U-bottom 96-well plate at 4°C, 50g for 2min. Aspirate 80uL of the supernatant with a pipette, prepare 10ug / mL Propidium iodide with PBS containing 2% FBS, add 100uL of PI reagent to each well, and stain for 15min. After staining, centrifuge the U-bottom 96-well plate at 4°C, 50g for 2min. Wash twice with PBS containing 2% FBS, add 100uL of PBS containing 2% FBS to resuspend, and detect by flow cytometry to count the average fluorescence intensity of Cy5 fluorescence of the living cell population.

[0496] 5. Data Analysis

[0497] The mean fluorescence intensity of Cy5 fluorescence in each experimental group was subtracted from the mean fluorescence intensity of the blank group, and the data were fitted with a binding saturation curve using GraphPadPrism software, as shown in Figure 1-3 As shown, the parameters were selected as Nonlinear regression (curvefit), One site binding (hyperbola), and the dissociation equilibrium constant Kd value was obtained. As shown in Table 3, the binding of G1-12, G4-12, and G5-12 to the ASGPR receptor all achieved high affinity.

[0498] Table 3 Kd values ​​of conjugates

[0499] sample G1-12 G4-12 G5-12 Bmax 153854 239297 199232 Kd 1.982 6.371 3.695

[0500] Experimental Example 2 In vivo liver targeting of GalNac-siRNA

[0501] Balb / c-nu mice, male, weighing 18-22g, were randomly divided into 3 groups, 5 mice / group, namely blank control group, negative control group and targeted group, and the dosage was 10mg / kg. The negative control group was injected with a negative control (NC, Ribobio, siM200922112438) test substance via the tail vein once, the targeted group was injected with the G1-12 test substance obtained in Example 6 via the tail vein once, and the blank control group was given an equal volume of normal saline. The livers of all animals were taken 6h after the drug and immediately placed in liquid nitrogen, then stored at -80℃, and the concentration of siRNA drugs in liver tissue was determined by qPCR.

[0502] 1. Prepare a standard curve. Refer to the instructions of the siRNA antisense strand standard (Ribobio, ssR190807024906) to prepare the standard sample stock solution, with an siRNA concentration of 500nM. Dilute the standard sample stock solution 10 times in a gradient (50nM~0.05pM), with a total of 7 gradients. Incubate at 95℃ for 10min, and quickly add pre-cooled RTmix for reverse transcription and qPCR. Reverse transcription refers to Bulge-Loop TM The miRNA qRT-PCR Starter Kit (Ribobio, C10211-2) was operated according to the instructions of the riboSCRIPTTM Reverse Transcription Kit (Ribobio, C11027-2). Based on the obtained Ct value, the average Ct value obtained for each standard point was calculated, and the log value of the standard point concentration was used as the horizontal axis and the average Ct value was used as the vertical axis to prepare a standard curve, and the regression equation and R were obtained. 2 , R 2 >0.99.

[0503] 2. Sample processing. Grind the mouse liver sample into powder using the liquid nitrogen grinding method and weigh the powder. Add the corresponding volume of 0.25% Triton X-100 (100 mg / ml) preheated at 95°C according to the mass and vortex. Mix well and incubate at 95°C for 10 minutes. Cool the lysate on ice and centrifuge at 20,000g at 4°C for 20 minutes. Collect the supernatant. Dilute the supernatant 10,000 times with RNase-free water, take 20 μl of the dilution and incubate it at 95°C in a PCR instrument for 10 minutes, and quickly add the pre-cooled RT mix for reverse transcription and qPCR. Refer to Bulge-Loop for reverse transcription. TM The miRNA qRT-PCR Starter Kit (Ribobio, C10211-2) was used for the operation, and the qPCR was performed according to the instruction manual of the riboSCRIPTTM Reverse Transcription Kit (Ribobio, C11027-2).

[0504] 3. Data analysis: Substitute the sample Ct value into the regression equation of the standard curve to calculate the siRNA content in the sample.

[0505] Depend on Figure 4 It can be seen that compared with the negative control siRNA without GalNAc, the distribution of G1-12 with GalNac-siRNA structure in mouse liver was significantly increased, indicating that G1-12 can achieve liver-targeted delivery in vivo.

[0506] Experimental Example 3: Knockdown of PCSK9 mRNA expression

[0507] Referring to Chinese invention patent CN109957567B, the synthetic siRNA sequence is as follows:

[0508] mG*mG*mUmCmUmGfGfAfAmUmGmCmAmAmAmG*mU*mC*mA (SEQ ID NO. 3),

[0509] mU*fG*mAfCmUfUmUfGmCfAmUfUmCfCmAfG*mA*mC*mC (SEQ ID NO. 4).

[0510] The compounds G2, G4 and G5 obtained in Examples 2, 4 and 5 were respectively connected to the above siRNA to form conjugates, as shown in Table 4.

[0511] Table 4 Conjugate structures

[0512]

[0513] Note: A is adenine ribonucleotide, G is guanine ribonucleotide, C is cytosine ribonucleotide, U is uracil ribonucleotide, f is 2'-fluoro modification, m is 2'-O-methyl modification, * is thiophosphate backbone modification, and 5'-phos* is thiophosphate at the 5' end.

[0514] Among them, the structure of G2-34 is shown in Z2. When G4 and G5 are connected to siRNA, the phosphate bond needs to be modified. In this embodiment, the amino group of C6 is used. Modification, so the structure of G4-34 is shown as Z4, and the structure of G5-34 is shown as Z5.

[0515] Hep3B cells (ATCC, HB-8064) were cultured, and the cell density before transfection was about 30%-50%, which was relatively uniform.

[0516] Each conjugate sample in Table 4 was prepared into a 20uM stock solution and set as the experimental group. In addition to the experimental group, a normal cell control group (Blank), a transfection reagent control group (Mock), a negative control group 1 (NC001, Ribobio, siM211215011852), and a negative control group 2 (NC002, Ribobio, siM211215011852) were set up. TM The cells were transfected according to the instruction manual of 3000 transfection reagent (Thermo Fisher, L300001). Both the experimental group and the control group were repeated 3 times.

[0517] Total RNA was extracted according to the instruction manual of MagZol Reagent (Magen, R4801).

[0518] Reverse transcription was performed with reference to the instruction manual of riboSCRIPTTM Reverse Transcription Kit (Ribobio, C11027-2).

[0519] The human housekeeping gene actin was used as the internal reference gene. The upstream primer sequence of the actin gene was 5'-TCAAGATCATTGCTCCTCCTGAG-3', and the downstream primer sequence was 5'-ACATCTGCTGGAAGGTGGACA-3'; the upstream primer sequence of the human PCSK9 gene was 5'-AAGCCAAGCCTCTTCTTACTTCA-3', and the downstream primer sequence was 5'-CCTGGGTGATAACGGAAAAAG-3'. Real-time fluorescence quantitative PCR reaction was performed using 2X SYBR Green Mix (Ribobio, C10712F-1ml) and a fluorescence quantitative PCR instrument (BioRad, CFX96). After the PCR reaction was completed, the Ct error of 9 replicates of a sample (3 transfection replicates × 3 QPCR replicates) should be within ±0.5. Then relative quantitative analysis was performed using CFX 2.1 software.

[0520] As shown in Table 5, the conjugates formed by different GalNAc structures and the same siRNA sequence have a knockdown efficiency of about 60% on the target gene PCSK9 in Hep3B cells, and can significantly knock down the expression of the target gene mRNA.

[0521] Table 5 PCSK9 mRNA relative expression values

[0522] Sample name Average relative expression NC001 1.00 NC002 0.87 Blank 0.98 Mock 0.97 G2-34 0.34 G4-34 0.41 G5-34 0.33

[0523] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A compound of formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof: in, M1 is independently selected from -O-, -S-, -NR6-, -CHR7-, -C(O)-, -S(O)- and -S(O)2-; R1 is an asialoglycoprotein receptor (ASGPR) ligand residue; R2 is selected from hydroxyl, carboxyl, -OR p 、-OPG2、 PG1 is a carboxyl protecting group; PG2 is a hydroxyl protecting group; R p for PG3 is independently selected from phosphate protecting groups; R and R' are each independently an amino protecting group; or R, R' and the N atom to which they are attached optionally together form a 5-10 membered heterocyclic group; R3 is independently M 3a Independently selected from chemical bonds, -O-, -S-, -NR6-, -CHR7-, -C(O)-, -S(O) 1-2 -, -NR6C(O)-, -C(O)NR6-, -NR6S(O) 1-2 -、-S(O) 1-2 NR6-, -OC(O)-, -C(O)O-, -OS(O) 1-2 -and-S(O) 1-2 O-; M 3b Independently selected from -O-, -S-, -NR6-, -CHR7-, -CH=CH-, -C≡C-, -C(O)-, -S(O) 1-2 -, -C(O)NR6-, -NR6C(O)-, -OC(O)-, -C(O)O-, -S(O) 1-2 O-Y1-、-OS(O) 1-2 -Y1-, -OS(O) 1-2 O-、-S(O) 1-2 NR6-Y1-、-NR6S(O) 1-2 -Y1-, C 6-10 Arylene, 5-10 membered heteroarylene, Y1 is independently C 6-10 Arylene or 5-10 membered heteroarylene; R6 are each independently selected from H, C 1-10 Alkyl, C 1-10 Haloalkyl, C 3-10 Cycloalkyl and 3-10 membered heterocyclic group; R7 are each independently selected from H, halogen, CN, NO2, C 1-10 Alkyl, C 1-10 Haloalkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic group, C 6-10 Aryl and 5-10 membered heteroaryl, wherein one or more methylene groups in the alkyl or haloalkyl are optionally and independently replaced by a group selected from the following: -C(O)-, -C(O)NH-, -NHC(O)-, -OC(O)-, -O-, -C(O)O-, -S-, -S-(CH2) 0-6 -S-、-NR6-、C 6-10 Arylene, 5-10 membered heteroarylene, -CH=CH-, -C≡C- and -O-(CH2) 1-6 -O-; n2 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; n3 is independently 1, 2, 3, 4, 5 or 6; R4 is selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl and C 1-6 Alkoxy; Link1, Link2, Link3 and Link4 are each independently C 1-25 Straight chain alkylene, wherein one or more methylene groups in the straight chain alkylene are optionally and independently replaced by R*, and R* is independently selected from: -O-, -S-, -NR6-, -CHR7-, -C(O)-, -S(O) 1-2 -, -NR6C(O)-, -C(O)NR6-, -NR6S(O) 1-2 -、-S(O) 1-2 NR6-, -OC(O)-, -C(O)O-, -OS(O) 1-2 -and-S(O) 1-2 O-; m1, m2 and m3 are each independently 1, 2, 3, 4 or 5; The condition is that the methylene group connected to the carbonyl group on its left side in Link4 is not replaced by R*.

2. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: M1 is independently selected from -O-, -S-, -NH-, -CH2-, -C(O)-, -S(O)- and -S(O)2-, preferably selected from -O-, -S- and -NH-, preferably -O-.

3. The compound of formula (I) according to claim 1 or 2, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: R1 is independently selected from the residues of the following structures: D-mannopyranose, L-mannopyranose, L-arabinose, D-arabinose, D-xylofuranose, L-xylofuranose, D-glucose, L-glucose, D-galactose, L-galactose, α-D-mannofuranose, β-D-mannofuranose, α-D-mannopyranose, β-D-mannopyranose, α-D-glucopyranose, β-D-glucopyranose, α- D-furanose, β-D-glucose, α-D-fructofuranose, β-D-fructofuranose, α-D-fructopyranose, β-D-fructopyranose, α-D-galactopyranose, β-D-galactopyranose, α-D-galactofuranose, β-D-galactofuranose, glucosamine, sialic acid, galactosamine, N-acetylgalactosamine, N-trifluoroacetylgalactosamine, N-propionylgalactosamine, N-butyrylgalactosamine, N-isobutyrylgalactosamine, 2-amino-3-O-[(R)-1-carboxyethyl]-2-deoxy-β-D-glucopyranose, 2-deoxy-2-methylamino-L-glucopyranose, 4,6-dideoxy-4-formamido-2,3-di-O-methyl-D-mannopyranose, 2-deoxy-2-sulfoamino-D-glucopyranose, N-glycolyl-α-neuraminic acid, 5-thio-β-D-glucopyranose glucose, 2,3,4-tri-O-acetyl-1-thio-6-O-trityl-α-D-glucopyranoside methyl ester, 4-thio-β-D-galactopyranose, 3,4,6,7-tetra-O-acetyl-2-deoxy-1,5-dithio-α-D-glucopyranoside ethyl ester, 2,5-anhydro-D-allosenitrile, ribose, D-ribose, D-4-thioribose, L-ribose and L-4-thioribose; Preferably, R1 is independently selected from the residues of the following structures: D-galactose, L-galactose, α-D-galactopyranose, β-D-galactopyranose, α-D-galactofuranose, β-D-galactofuranose, galactosamine, N-acetylgalactosamine, N-trifluoroacetylgalactosamine, N-propionylgalactosamine, N-n-butyrylgalactosamine, N-isobutyrylgalactosamine and β-D-galactopyranose, preferably selected from: galactosamine, N-acetylgalactosamine, N-trifluoroacetylgalactosamine, N-propionylgalactosamine, N-n-butyrylgalactosamine and N-isobutyrylgalactosamine, preferably N-acetylgalactosamine; Preferably, R1 is 4. A compound of formula (I) according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: R2 is selected from hydroxyl, carboxyl, -OR p 、-OPG2、 Preferably selected from -OR p , Preferably selected from Preferably selected from Preferably Preferably Preferably, for Preferably 5. The compound of formula (I) according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein PG 1 from C 1-6 Alkyl, benzyl, allyl, Among them, R 2s are independently selected from H, halogen, C 1-10 Alkyl and C 1-10 haloalkyl, q is 0, 1, 2, 3, 4 or 5; Preferably, PG1 is selected from methyl, tert-butyl, benzyl, allyl, Preferably selected from Preferably Preferably Preferably, R 2s are independently selected from H, halogen, C 1-6 Alkyl and C 1-6 The haloalkyl group is preferably H or halogen, preferably F.

6. A compound of formula (I) according to any one of claims 1 to 5, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: PG2 is selected from: trimethylsilyl (TMS), triethylsilyl (TES), dimethylisopropylsilyl (DMIPS), diethylisopropylsilyl (DEIPS), tert-butyldimethylsilyl ether (TBDMS), tert-butyldiphenylsilyl (TBDPS), triisopropylsilyl ether (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, dimethoxytrityl (DMT), monomethoxytrityl (MMT), 9-phenylxanthen-9-yl (Pixyl), 9-(p-methoxyphenyl)xanthen-9-yl (Mox), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr), 4,4',4"-trimethoxytrityl (TMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), and p-methoxybenzyloxymethyl (PMBM); Preferably, PG2 is selected from dimethoxytrityl (DMT), monomethoxytrityl (MMT), 9-phenylxanthen-9-yl (Pixyl), 9-(p-methoxyphenyl)xanthen-9-yl (Mox), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr), 4,4',4"-trimethoxytrityl (TMTr), tert-butyldimethylsilyl ether (TBMDS), tert-butyldiphenylsilyl (TBDPS) and triisopropylsilyl ether (TIPS), preferably selected from dimethoxytrityl (DMT), monomethoxytrityl (MMT), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr) and 4,4',4"-trimethoxytrityl (TMTr), preferably 4,4'-dimethoxytrityl (DMTr).

7. A compound of formula (I) according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein PG 3 is independently selected from 2-cyanoethyl and C 1-6 The alkyl group is preferably selected from 2-cyanoethyl and methyl, and is preferably 2-cyanoethyl.

8. A compound of formula (I) according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: R and R' are each independently C 1-6 The alkyl group is preferably an isopropyl group.

9. A compound of formula (I) according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: R, R' and the nitrogen atom to which they are attached are optionally taken together to form a 5-6 membered heterocyclic group.

10. A compound of formula (I) according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: R p for 11. A compound of formula (I) according to any one of claims 1 to 10, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: M 3a Independently selected from chemical bonds, -O-, -S-, -NR6-, -CHR7-, -C(O)-, -S(O) 1-2 -, -NHC(O)-, -C(O)NH-, -NHS(O) 1-2 -、-S(O) 1-2 NH-, -OC(O)-, -C(O)O-, -OS(O) 1-2 -and-S(O) 1-2 O-, preferably selected from a chemical bond, -O-, -S-, -NR6-, -C(O)-, -NHC(O)-, -C(O)NH-, -OC(O)- and -C(O)O-, preferably selected from a chemical bond, -NHC(O)-, -C(O)NH-, -OC(O)- and -C(O)O-, preferably selected from a chemical bond, -NHC(O)- and -C(O)NH-, preferably a chemical bond or -C(O)NH-.

12. A compound of formula (I) according to any one of claims 1 to 11, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: M 3b Independently selected from -O-, -S-, -NR6-, -CHR7-, -CH=CH-, -C≡C-, -C(O)-, -S(O) 1-2 -, -C(O)NH-, -NHC(O)-, -OC(O)-, -C(O)O-, -S(O) 1-2 O-Y1-、-OS(O) 1-2 -Y1-, -OS(O) 1- 2O-、-S(O) 1-2 NH-Y1-, -NHS(O) 1-2 -Y1-, phenylene, 5-6 membered heteroarylene, Preferably, M 3b independently selected from -O-, -S-, -NR6-, -CHR7-, -CH=CH-, -C≡C-, -C(O)-, -C(O)NH-, -NHC(O)-, -OC(O)-, -C(O)O-, -OS(O)2-Y1-, -OS(O)2O-, -NHS(O)2-Y1-, 5-6 membered heteroarylene, It is preferably selected from -C(O)NH-, -NHC(O)-, -OC(O)-, -C(O)O-, preferably -C(O)NH- or -NHC(O)-, preferably -NHC(O)-; Preferably, M 3b The 5-6 membered heteroarylene group is Preferably, Y1 is independently phenylene or 5-6 membered heteroarylene, preferably phenylene, preferably 13. A compound of formula (I) according to any one of claims 1 to 12, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: Selected from the following structures: Preferably it is selected from A2, A3, A4, A6 and A9, preferably selected from A2, A3 and A9, preferably A3 or A9.

14. A compound of formula (I) according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: R6 are each independently selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-7 Cycloalkyl and 3-7 membered heterocyclic groups, preferably selected from H, C 1-6 Alkyl and C 1-6 Haloalkyl, preferably selected from H, -CH3, More preferably, it is H.

15. A compound of formula (I) according to any one of claims 1 to 14, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: R7 are each independently selected from H, halogen, CN, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-7 Cycloalkyl, 3-7 membered heterocyclyl, phenyl and 5-6 membered heteroaryl, preferably selected from H, halogen, CN, C 1-6 Alkyl and C 1-6 Haloalkyl, preferably selected from H, halogen, C 1-6 Alkyl and C 1-6 Haloalkyl; Preferably, one or more methylene groups in the alkyl or haloalkyl in R7 are optionally and independently replaced by a group selected from the following: -C(O)-, -C(O)NH-, -NHC(O)-, -OC(O)-, -O-, -C(O)O-, -S-, -S-(CH2) 0-3 -S-, -NR6-, phenylene, 5-6 membered heteroarylene, -CH=CH-, -C≡C- and -O-(CH2) 1-3 -O- is preferably selected from -C(O)-, -C(O)NH-, -NHC(O)-, -OC(O)-, -O-, -C(O)O-, -S-, -SS-, -NR6-, -CH=CH-, -C≡C- and -O-(CH2)2-O-; Preferably, the halogen in R7 is F, Cl or Br.

16. A compound of formula (I) according to any one of claims 1 to 15, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: n2 is independently 0, 1, 2, 3, 4, 5 or 6, preferably 0, 1, 2 or 3, preferably 0 or 3.

17. A compound of formula (I) according to any one of claims 1 to 16, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: n3 is independently 1, 2 or 3, preferably 2.

18. A compound of formula (I) according to any one of claims 1 to 17, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: R4 is selected from H, C 1-5 Alkyl, C 1-5 Haloalkyl and C 1-5 Alkoxy, preferably selected from H, C 1-3 Alkyl and C 1-3 Halogenated alkyl, preferably H.

19. A compound of formula (I) according to any one of claims 1 to 18, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein Li nk1, Link2 and Link3 are each independently C 1-10 Straight chain alkylene, preferably C 1-6 Straight chain alkylene, preferably C 4-6 The straight chain alkylene group is preferably -(CH2)4- or -(CH2)6-.

20. A compound of formula (I) according to any one of claims 1 to 19, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein Li nk4 is C 1-20 Straight chain alkylene, preferably C 1-17 Straight chain alkylene, preferably C 1-10 Straight chain alkylene, preferably C 3-20 Straight chain alkylene, preferably C 3-17 Straight chain alkylene, preferably C 3-10 Straight chain alkylene; Preferably, one or more methylene groups, preferably 1, 2 or 3 methylene groups, preferably 1 methylene group in the linear alkylene group in Link4 are optionally and independently replaced by R*; Preferably, R* is independently selected from: -O-, -NR6-, -CHR7-, -C(O)-, -NR6C(O)-, -C(O)NR6-, -OC(O)- and -C(O)O-, preferably -C(O)NH-, -NHC(O)-, -C(O)O- and -OC(O)-, preferably -C(O)NH- and -C(O)O-, preferably -C(O)NH- and -NHC(O)-, preferably -C(O)NH-; Preferably, R* is separated from the carbonyl group on the left side of Link4 by 1-15 methylene groups, preferably 1-12 methylene groups, preferably 1-10 methylene groups, preferably 1-3 methylene groups, preferably 3-10 methylene groups; R and R2 are separated by 1-12 methylene groups, preferably 1-10 methylene groups, preferably 1-6 methylene groups; Preferably, Link4 is selected from -(CH2)3-, -(CH2) 10 -, -(CH2)3-C(O)NH-(CH2)6- and -(CH2) 10 -C(O)NH-(CH2)6-, preferably selected from -(CH2)3-, -(CH2) 10 - and -(CH2)3-C(O)NH-(CH2)6-, preferably -(CH2)3- or -(CH2) 10 -, preferably selected from -(CH2) 10 -, -(CH2)3-C(O)NH-(CH2)6- and -(CH2) 10 -C(O)NH-(CH2)6-, preferably -(CH2) 10 -or-(CH2) 10 -C(O)NH-(CH2)6-.

21. A compound of formula (I) according to any one of claims 1 to 20, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: m1, m2 and m3 are each independently 1, 2 or 3, preferably 1.

22. A compound of formula (I) according to any one of claims 1 to 21, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, having the following structural formula: in, n2 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; The remaining groups are as defined in any one of claims 1 to 21.

23. The compound of formula (II) according to claim 22, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: R2 is selected from hydroxyl, carboxyl, -OR p 、-OPG2、 PG1 is a carboxyl protecting group, preferably selected from methyl, tert-butyl, benzyl, allyl, R 2s are independently selected from H, halogen, C 1-6 Alkyl and C 1-6 Haloalkyl; q is 0, 1, 2, 3, 4, or 5; PG2 is a hydroxy protecting group, preferably selected from dimethoxytrityl (DMT), monomethoxytrityl (MMT), 9-phenylxanthen-9-yl (Pixyl), 9-(p-methoxyphenyl)xanthen-9-yl (Mox), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr), 4,4',4"-trimethoxytrityl (TMTr), tert-butyldimethylsilyl ether (TBMDS), tert-butyldiphenylsilyl (TBDPS) and triisopropylsilyl ether (TIPS); R p for PG3 is independently selected from phosphate protecting groups, preferably selected from 2-cyanoethyl and methyl, more preferably 2-cyanoethyl; R and R' are each independently an amino protecting group, preferably C 1-6 Alkyl, preferably isopropyl; M 3a is selected from the group consisting of a chemical bond, -NHC(O)-, -C(O)NH-, -OC(O)- and -C(O)O-, preferably selected from the group consisting of a chemical bond, -NHC(O)- and -C(O)NH-; n1 is independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; n2 is independently 0, 1, 2, 3, 4, 5 or 6; n3 is independently 1, 2, 3, 4, 5 or 6; m1, m2 and m3 are each independently 1, 2, 3, 4 or 5; Link4 is selected from C 1-25 Straight chain alkylene, preferably C 1-20 Straight chain alkylene, wherein 1, 2 or 3 methylene groups in the straight chain alkylene are optionally and independently replaced by R*, and R* is independently selected from: -C(O)NH-, -NHC(O)-, -C(O)O- and -OC(O)-, preferably -C(O)NH- and -NHC(O)-; The condition is that the methylene group connected to the carbonyl group on its left side in Link4 is not replaced by R*.

24. The compound of formula (II) according to claim 23, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: R2 is selected from -OR p , Preferably selected from Preferably PG1 is selected from R 2s are independently H or halogen, preferably F; q is 0, 1, 2, 3, 4, or 5; PG2 is selected from dimethoxytrityl (DMT), monomethoxytrityl (MMT), trityl (Tr), 4-methoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr) and 4,4',4"-trimethoxytrityl (TMTr), preferably 4,4'-dimethoxytrityl (DMTr); R p for PG3 is 2-cyanoethyl; R and R' are each independently C 1-4 Alkyl, preferably isopropyl; M 3a is a chemical bond or -C(O)NH-; n1 is independently 4, 5 or 6; n2 is independently 0, 1, 2 or 3; n3 is independently 1 or 2; m1, m2 and m3 are each independently 1, 2 or 3, preferably 1; Link4 is selected from C 1-17 Straight chain alkylene, preferably selected from C 1-10 Straight chain alkylene, wherein one methylene in the straight chain alkylene is optionally replaced by R*, and R* is -C(O)NH-; Preferably, R* is separated from the carbonyl group on the left side of Link4 by 1-12 methylene groups, preferably 1-10 methylene groups, preferably 1-3 methylene groups; R* is separated from R2 by 1 to 12 methylene groups, preferably 1 to 10 methylene groups, and preferably 1 to 6 methylene groups.

25. The compound of formula (II) according to claim 24, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: R2 is selected from Preferably M 3a is a chemical bond or -C(O)NH-; n1 is independently 4 or 6; n2 is independently 0 or 3; n3 is 2; m1, m2 and m3 are 1; Link4 is selected from -(CH2)3-, -(CH2) 10 -、-(CH2)3-C(O)NH-(CH2)6- or -(CH2) 10 -C(O)NH-(CH2)6-, preferably -(CH2)3-, -(CH2) 10 - and -(CH2)3-C(O)NH-(CH2)6-, preferably -(CH2)3- or -(CH2) 10 -.

26. The compound of formula (I) of claim 1, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: The compound is selected from the following:

27. A conjugate having a structure represented by formula (III): in, L is a structure represented by formula (I') or formula (II'): Selected from Preferably selected from Preferably selected from Preferably selected from Preferably selected from R D is a drug molecule independently selected from a cytotoxic agent, a chemotherapeutic agent, a somatostatin, a toxin, a radioisotope, and an oligonucleotide chain; d is 1 or 2, preferably 1; The remaining groups are as defined in any one of claims 1 to 25; Preferably, Link4 is selected from C 5-25 Straight chain alkylene, preferably C 5-20 Straight chain alkylene, preferably C 8-20 Straight chain alkylene, preferably C 10-17 Straight chain alkylene; Preferably, one or more methylene groups, preferably 1, 2 or 3 methylene groups, preferably 1 methylene group in Link4 are optionally and independently replaced by R*, R* being independently selected from: -C(O)NH-, -NHC(O)-, -C(O)O- and -OC(O)-, preferably -C(O)NH- and -C(O)O-, preferably -C(O)NH- and -NHC(O)-, preferably -C(O)NH-; Preferably, R* is separated from the carbonyl group on the left side of Link4 by 1-15 methylene groups, preferably by 1-12 methylene groups, preferably by 1-10 methylene groups, preferably by 3-10 methylene groups; R* and R2 are separated by 1-12 methylene groups, preferably 1-10 methylene groups, preferably 1-6 methylene groups; Preferably, Link4 is selected from -(CH2) 10 -, -(CH2)3-C(O)NH-(CH2)6- and -(CH2) 10 -C(O)NH-(CH2)6-, preferably -(CH2) 10 -or-(CH2) 10 -C(O)NH-(CH2)6-.

28. The conjugate of formula (III) according to claim 27, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: R D is an oligonucleotide chain, optionally, the oligonucleotide chain is a single-stranded oligonucleotide or a double-stranded oligonucleotide or a combination thereof; Preferably, M2 is linked to the 5' end, 3' end or any nucleotide between the 5' end and the 3' end of at least one chain of the oligonucleotide chain through a phosphate bond, preferably linked to the 5' end or the 3' end of at least one chain, preferably linked to the 5' end of at least one chain, and preferably linked to the 3' end of at least one chain; Preferably, M2 is linked to the sense strand of the oligonucleotide strand via a phosphate bond; Preferably, the phosphate bond is a phosphodiester bond or a modified phosphate bond, preferably a phosphodiester bond; preferably, the modified phosphate bond is selected from a thio-modified phosphate bond and an amino-modified phosphate bond, preferably a thio-modified phosphate bond; Preferably, the Selected from the following structures: Preferably selected from Preferably selected from in, represents an oligonucleotide chain.

29. The conjugate of formula (III) according to claim 27 or 28, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: The oligonucleotide chain includes unmodified nucleotides and / or modified nucleotides; Preferably, the modified nucleotides are each independently selected from 2′-methoxyethyl modified nucleotides, 2′-O-alkyl modified nucleotides, 2′-O-allyl modified nucleotides, 2′-C-allyl modified nucleotides, 2′-fluoro modified nucleotides, 2′-deoxy modified nucleotides, 2′-hydroxy modified nucleotides, phosphorothioate backbone modified nucleotides, locked nucleotide modified nucleotides, glycerol nucleic acid (glycol nucleic acid, GNA) modified nucleotides and unlocked nucleic acid (unlocked nucleic acid, UNA) modified nucleotides, preferably selected from 2′-O-alkyl modified nucleotides, 2′-fluoro modified nucleotides, 2′-deoxy modified nucleotides and phosphorothioate backbone modified nucleotides; Preferably, the 2′-O-alkyl group is 2′-OC 1-6 Alkyl, preferably 2′-O-methyl; Preferably, the length of the oligonucleotide chain is 5-100 bp.

30. The conjugate of formula (III) according to any one of claims 27 to 29, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: The oligonucleotide chain is siRNA, ASO or microRNA, preferably siRNA.

31. The conjugate of formula (III) according to any one of claims 27 to 30, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: The oligonucleotide chain has a terminal modification, and the terminal modification is selected from cholesterol, polyethylene glycol, a fluorescent probe, biotin, a polypeptide, a vitamin, a tissue targeting molecule, or a combination thereof.

32. The conjugate of formula (III) according to any one of claims 27 to 31, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: The ribose-phosphate backbone of the oligonucleotide chain is replaced by polypeptide nucleic acid (PNA) or morpholino antisense nucleotide (PMO).

33. The conjugate of formula (III) according to any one of claims 27 to 32, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: L is selected from the following structures:

34. The conjugate of formula (III) according to claim 28, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein: The conjugate is selected from the following:

35. A pharmaceutical composition comprising the conjugate of any one of claims 27 to 34, and optionally a pharmaceutically acceptable carrier, excipient, adjuvant or vehicle, and optionally other therapeutic agents.

36. Use of the conjugate according to any one of claims 27 to 34, or the pharmaceutical composition according to claim 35, in the preparation of a medicament for treating or preventing a disease associated with the expression or overexpression of a gene in hepatocytes.

37. A method for treating or preventing a disease associated with expression or overexpression of a gene in hepatocytes in a subject, comprising administering to the subject the conjugate of any one of claims 27 to 34 or the pharmaceutical composition of claim 35.

38. The conjugate according to any one of claims 27 to 34 or the pharmaceutical composition according to claim 35, for use in treating or preventing a disease associated with the expression or overexpression of a gene in hepatocytes.

39. The use of claim 36 or the method of claim 37 or the conjugate or pharmaceutical composition of claim 38, wherein the gene is selected from HBV genome, HCV genome, PCSK9, xanthine oxidase, URAT1, APOB, liver fibrosis-related genes (AP3S2, AQP2, AZINI, DEGSI, STXBP5L, TLR4, TRPM5), non-alcoholic fatty liver disease-related genes (PNPLA3, FDFT1), primary biliary cirrhosis-related genes (HLA-DQB1, IL-12, IL-12RB2), or a combination thereof.

40. The use of claim 36 or the method of claim 37 or the conjugate or pharmaceutical composition of claim 38, wherein the disease is selected from: hereditary angioedema, familial tyrosinemia type I, Alagille syndrome, alpha-1-antitrypsin deficiency, defects in bile acid synthesis and metabolism, biliary atresia, cystic fibrosis liver disease, idiopathic neonatal hepatitis, mitochondrial liver disease, progressive familial intrahepatic cholestasis, primary sclerosing cholangitis, transthyretin amyloidosis, hemophilia, homozygous familial hypercholesterolemia, hyperlipidemia, hepatitis B (HBV), hepatitis C (HCV), steatohepatitis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), hyperglycemia or diseases involving abnormally increased hepatic glucose production similar to type III diabetes, hepatitis and hepatic porphyrins.

41. Compounds of formula (C): in, PG2 and PG4 are protecting groups; Preferably, PG2 is a hydroxyl protecting group and PG4 is an amino protecting group; Preferably, the PG2 is selected from trimethylsilyl (TMS), triethylsilyl (TES), dimethylisopropylsilyl (DMIPS), diethylisopropylsilyl (DEIPS), tert-butyldimethylsilyl ether (TBDMS), tert-butyldiphenylsilyl (TBDPS) and triisopropylsilyl ether (TIPS), preferably tert-butyldimethylsilyl ether (TBDMS); Preferably, the PG4 is selected from benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), 9-fluorenylmethoxycarbonyl (Fmoc), allyloxycarbonyl (Alloc), 2-(trimethylsilyl)ethoxycarbonyl (Teoc), methoxycarbonyl and ethoxycarbonyl, preferably tert-butyloxycarbonyl (Boc).

42. A method for preparing a compound of formula (C), comprising the steps of: reacting compound A with compound B to generate compound C: in, PG2 and PG4 are as defined in claim 41; Preferably, the reaction conditions of the method include a condensing agent and a base; Preferably, the condensing agent is selected from carbodiimide condensing agents, onium salt condensing agents and organophosphorus condensing agents; Preferably, the reaction conditions further include a condensation activator; Preferably, the reaction conditions further include a solvent; Preferably, the method is carried out in a nitrogen atmosphere; Preferably, the base is an organic amine base, preferably DIEA, TEA, NMM, DBU, DMAP, pyridine, 2,6-lutidine and 2,2,6,6-tetramethylguanidine, preferably DIEA.

43. Compounds of formula (D):

44. A method for preparing a compound of formula (D), comprising the steps of: removing the protecting group from the compound C of claim 41 to obtain a compound D: in, PG2 and PG4 are as defined in claim 41; Preferably, the method is carried out under acidic conditions; Preferably, the reaction conditions of the method include an acid and a solvent; Preferably, the acid is HCl and / or H2SO4, preferably HCl; Preferably, the solvent is an organic solvent, preferably an alcohol solvent, preferably selected from methanol, ethanol, propanol, butanol, isopropanol, cyclohexanol, 2-methyl-2-propanol and 2-ethyl-1-propanol, preferably methanol.

Images