Conjugate and use thereof

Abstract

The present application relates to a conjugate and a use thereof. The conjugate comprises a linker unit L or a pharmaceutically acceptable salt thereof, and the structural general formula thereof is as shown in formula (I-L).

Description

A conjugate and its application Technical Field

[0001] This application relates to the field of biomedicine, specifically to a conjugate and its applications. Background Technology

[0002] Currently, nucleic acid drugs, especially oligonucleotide drugs, are widely used due to their simple synthesis and high activity. Oligonucleotide drugs typically include antisense oligonucleotides (ASO), small interfering RNA (siRNA), microRNA (miRNA), and nucleic acid aptamers.

[0003] N-acetylgalactosamine (GalNAc) is a ligand that binds to the desialization glycoprotein receptor (ASGPR) on the liver surface. The desialization glycoprotein receptor is an endocytic receptor specifically expressed on the surface of hepatocytes. In recent years, the high-affinity ligand GalNAc, used as a targeting molecule, has made some progress in the liver-targeted delivery of nucleic acid drugs. For example, Alnylam Pharmaceuticals, Inc. reported that siRNA based on GalNAc conjugation technology exerted gene silencing activity in mice (Nair JK, et al. J. Am. Chem. Soc. 2014, 136, 16958). The paper reported that the GalNAc-siRNA conjugate exhibited good delivery activity in both in vivo and in vitro experiments. In in vivo experiments in mice administered subcutaneously, the ED50 for a single dose was determined to be 1 mg / kg, with a single injection dose of less than 1 mL. In long-term dosing studies, weekly subcutaneous injections resulted in stable interference activity for up to 9 months. The study found that GalNAc compounds with four or three antennae have a much higher affinity for ASGPR than GalNAc compounds with two or one antennae.

[0004] Different ligand structures result in significantly different nucleic acid delivery effects. There remains a need in this field to develop new ligand compounds. Summary of the Invention

[0005] This application constructs linker units (e.g., IL, IW) based on 3,4-position substituted pyrrole rings for linking nucleotide drugs (e.g., siRNA) and ligands (e.g., GalNAc).

[0006] In some embodiments, nucleotide drug delivery systems (e.g., IX) or oligonucleotide conjugates formed with this linker unit have liver-targeted delivery efficiency comparable to or better than existing structurally similar GalNAc compounds.

[0007] In some embodiments, nucleotide drug delivery systems (e.g., IX) or oligonucleotide conjugates formed with this linker unit have comparable or better long-lasting effects compared to existing structurally similar GalNAc compounds.

[0008] In some embodiments, nucleotide drug delivery systems (e.g., IX) or oligonucleotide conjugates formed with this linker unit require an effective dose that is comparable to or lower than that of existing structurally similar GalNAc compounds.

[0009] In some embodiments, nucleotide drug delivery systems (e.g., IX) or oligonucleotide conjugates formed from this linker unit are easier and more economical to synthesize than existing structurally similar GalNAc compounds, and are suitable for scale-up synthesis and production.

[0010] On the one hand, this application provides a linker unit L or a pharmaceutically acceptable salt thereof, the general structural formula of which is shown in formula (IL):

[0011] in,

[0012] Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group;

[0013] Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; X1 is a branched chain; X2 is a nucleophilic or electrophilic group; R1-R6 are any substituents.

[0014] In some embodiments, the connector unit L further includes W, whose molecular formula is shown as (IW).

[0015] W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino-substituted group or amino-protecting group optionally substituted with one or more halogens.

[0016] In some embodiments, the linker unit L further includes a ligand X, the molecular formula of which is shown in (IX):

[0017] Where X is a ligand or a derivative thereof.

[0018] In some embodiments, R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3alkyl, optional substituent C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 The group may be cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; wherein the optional substituent group may be halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

[0019] In some embodiments, X1 includes -(CH2) n -、-(CH2) n NH-, -(CH2) n O-, -(CH2) n S-、-R X12 (CH2) n -、-R X12 (CH2) n NH-, -R X12 (CH2) n O- or -R X12 (CH2) n One of S-; where R X12 It is one of N, O, and S; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0020] In some embodiments, X1 includes -(CH2) n -、-(CH2) n NH-, -(CH2) n O-, -(CH2) n S-、-R X12 (CH2) n -、

[0021] -R X12 (CH2) n N-、-R X12 (CH2) n O- or -R X12 (CH2) n Optional substitutions for S-;

[0022] The substituents optionally substituted may be selected from the group consisting of: halogen, cyano, hydroxy, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C3 alkyl polyoxyethylene, or C1-C3 alkyl polyoxypropylene;

[0023] Where R X12 It is one of N, O, and S;

[0024] n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0025] 39. The compound according to any one of claims 1-6, wherein,

[0026] X1 includes -((CH2) m1 O) m2 -,-((CH2) m1 O) m2 CH2-, -CH2((CH2) m1 O) m2 CH2-

[0027] m1 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5;

[0028] m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5.

[0029] In some embodiments, X1 includes -((CH2) m1 O) m2 -,-((CH2) m1 O) m2 CH2-, -CH2((CH2) m1 O) m2 Optional substitution of CH2-, wherein the substituent group of the optionally substituted group is selected from the group consisting of: halogen, cyano, hydroxy, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C3 alkyl polyoxyethylene, or C1-C3 alkyl polyoxypropylene; wherein R X12 It is one of N, O, and S; m1 is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0030] In some embodiments, the compound or a pharmaceutically acceptable salt thereof, as shown in formula (IL), formula (IW), or formula (IX), further comprises one of the following structures:

[0031] Where R1-R 12 Including hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-6 alkyl, optional substituent C 1-6 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-6 alkyl carbonyl, C with optional substituent 1-6 One of cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent;

[0032] The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

[0033] n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5;

[0034] m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5;

[0035] Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group;

[0036] Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group;

[0037] X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-;

[0038] X is a ligand or a derivative thereof;

[0039] W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino substituent, or amino protecting group optionally substituted with one or more halogens.

[0040] In some embodiments, X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-.

[0041] In some embodiments, the compound of formula (IL), formula (IW), or formula (IX), or a pharmaceutically acceptable salt thereof, further has one of the following structures:

[0042] Where R1-R 12 Including hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-6 alkyl, optional substituent C 1-6 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-6 alkyl carbonyl, C with optional substituent 1-6 One of cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent;

[0043] The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

[0044] W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino substituent or amino protecting group optionally substituted by one or more halogens;

[0045] X is a ligand or a derivative thereof;

[0046] n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5;

[0047] m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5;

[0048] Y includes hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, and -P(R Y1 2. -P(OR) Y1 )(N(R Y1 )2), -P(=O)(OR Y1 )R Y1 -P(=S)(OR Y1 )R Y1 -P(=O)(SR) Y1 )R Y1 -P(=S)(SR) Y1 )R Y1 -P(=O)(OR) Y1 )2、-P(=S)(OR Y1 )2、-P(=O)(SR Y1 )2、-P(=S)(SR Y1 )2、-P(=O)(NHSO2CH3)R Y1 One of the substituted acyl groups, wherein each R Y1 Independently hydrogen, oxygen, hydroxyl, or a C1-C6 alkyl group optionally substituted with one or more halogens or cyano groups;

[0049] Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group;

[0050] W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino-substituted group or amino-protecting group optionally substituted with one or more halogens.

[0051] In some embodiments, the compound of formula (IL), formula (IW) or formula (IX) or a pharmaceutically acceptable salt thereof further has one of the following structures:

[0052] Z comprises one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; X1 is branched; X2 comprises one of -NH2, -SH, -OH, -NH-, -O-, or -S-; W comprises hydrogen, amino, hydroxyl, thiol group, a C1-C6 alkyl group optionally substituted with one or more halogens, an amino substituent group, or an amino protecting group; n is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5; m2 is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5; wherein R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 The group may be cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; wherein the optional substituent group may be halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

[0053] In some embodiments, the compound of formula (IL), formula (IW) or formula (IX) or a pharmaceutically acceptable salt thereof further has one of the following structures:

[0054] Y comprises one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; X1 is branched; X2 comprises one of -NH2, -SH, -OH, -NH-, -O-, or -S-; W comprises hydrogen, amino, hydroxyl, thiol group, a C1-C6 alkyl group optionally substituted with one or more halogens, an amino substituent group, or an amino protecting group; n is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5; m2 is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5; wherein R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 The group may be cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; wherein the optional substituent group may be halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

[0055] In some embodiments, the compound of formula (IL), formula (IW) or formula (IX) or a pharmaceutically acceptable salt thereof further has one of the following structures:

[0056] Y comprises one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; Z comprises one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; X1 is branched; X2 comprises one of -NH2, -SH, -OH, -NH-, -O-, or -S-; W comprises hydrogen, amino, hydroxyl, thiol group, a C1-C6 alkyl group optionally substituted with one or more halogens, an amino-substituted group, or an amino protecting group; n is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5; m2 is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5; wherein, R5-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional substituent C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 The group may be cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; wherein the optional substituent group may be halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

[0057] In some embodiments, X includes galactose, galactosamine, N-acetylgalactosamine, mannose, glucose, glucosamine, N-acetylglucosamine, fucose or lactose, N-acetylgalactosamine with hydroxyl groups fully protected by an acyl group, galactose with hydroxyl groups fully protected by an acyl group, galactosamine with hydroxyl groups fully protected by an acyl group, N-formyl-galactosamine with hydroxyl groups fully protected by an acyl group, N-propionyl-galactosamine with hydroxyl groups fully protected by an acyl group, N-butyryl-galactosamine with hydroxyl groups fully protected by an acyl group, or N-isobutyryl-galactosamine with hydroxyl groups fully protected by an acyl group, wherein the acyl group is an acetyl group or a benzoyl group.

[0058] In some embodiments, X is a lipid or a lipid-based molecule.

[0059] In some embodiments, the molecular formula of X is selected from one of the following:

[0060] Where r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0061] In some embodiments, the compound of formula (IX) or a pharmaceutically acceptable salt thereof further has one of the following structures.

[0062] Y comprises one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; Z comprises one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; X1 is branched; X2 comprises one of -NH2, -SH, -OH, -NH-, -O-, or -S-; n is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5; m2 is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5; r is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5; wherein R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 The group may be cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; wherein the optional substituent group may be halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

[0063] In some embodiments, the compound of formula (IW) or formula (IX) or a pharmaceutically acceptable salt thereof further has the following general formula:

[0064] Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group;

[0065] Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group;

[0066] n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5;

[0067] X is a ligand or a derivative thereof;

[0068] W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino substituent, or amino protecting group optionally substituted with one or more halogens.

[0069] In some embodiments, the compound of formula (IW) or a pharmaceutically acceptable salt thereof further has one of the following structures:

[0070] Y comprises one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; Z comprises one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; n is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5; m2 is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5; r is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5;

[0071] Among them, R5-R 12 Selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional substituent C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 Cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent;

[0072] The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

[0073] In some embodiments, the compound of formula (IX) or a pharmaceutically acceptable salt thereof further has one of the following structures.

[0074] Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group;

[0075] Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group;

[0076] n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5;

[0077] m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5;

[0078] r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0079] On the other hand, this application provides a support structure, including the connector unit L as described in claims 1-21, comprising one of the following structures:

[0080] Where q is an integer from 1 to 3.

[0081] Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group;

[0082] Z comprises one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; X1 is branched; X2 is a nucleophilic or electrophilic group; X2 comprises one of -NH2, -SH, -OH, -NH-, -O-, or -S-; W comprises hydrogen, amino, hydroxyl, thiol group, a C1-C6 alkyl group optionally substituted with one or more halogens, an amino-substituted group, or an amino protecting group; wherein, R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 The group may be cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; wherein the optional substituent group may be halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

[0083] In some embodiments, the support structure further includes one of the following structures:

[0084] X1 is a branched chain; X2 is a nucleophilic or electrophilic group; X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-; W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino-substituted, or amino-protecting groups optionally substituted with one or more halogens; wherein R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent1-5 The group may be cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; wherein the optional substituent group may be halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

[0085] In some embodiments of the support structure, the following general formula is used:

[0086] X1 is a branch;

[0087] X2 is a nucleophilic or electrophilic group; X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-;

[0088] W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino-substituted group or amino-protecting group optionally substituted with one or more halogens.

[0089] n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0090] m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5.

[0091] r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0092] Among them, R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 Cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent;

[0093] The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

[0094] In some embodiments, the scaffold structure further includes one having the following chemical formula:

[0095] n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0096] In some embodiments, the support structure further has one of the following structures:

[0097] n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0098] On the other hand, this application provides a conjugate, wherein the conjugate comprises: (i) one or more oligonucleotides; (ii) one or more ligands X; and (iii) one or more linker units L, wherein the conjugate structure comprises one of the following structures:

[0099] Where X is a ligand or its derivative, and RA is an oligonucleotide.

[0100] On the other hand, this application provides a conjugate with the following general structural formula:

[0101] Wherein, L is the linker unit L as described in any one of claims 1-21; X is a ligand or a derivative thereof; W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl optionally substituted with one or more halogens, amino substituents or amino protecting groups; RA is an oligonucleotide; p is 0-10, preferably 1-3, more preferably 2.

[0102] In some embodiments, the conjugate further has one of the following structures:

[0103] X1 is a branch;

[0104] X2 is a nucleophilic or electrophilic group; X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-;

[0105] W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino-substituted group or amino-protecting group optionally substituted with one or more halogens.

[0106] n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0107] m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5.

[0108] r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0109] Among them, R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 Cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent;

[0110] The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

[0111] In some embodiments, the conjugate further comprises one having the following chemical formula:

[0112] n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5;

[0113] m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5;

[0114] r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0115] RA stands for oligonucleotide.

[0116] In some embodiments, the conjugate further has one of the following structures:

[0117] n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5;

[0118] m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5;

[0119] r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0120] RA stands for oligonucleotide.

[0121] In some embodiments, the oligonucleotide includes a single-stranded oligonucleotide or a double-stranded oligonucleotide.

[0122] In some embodiments, the oligonucleotide comprises one or more modified nucleotides.

[0123] In some embodiments, the one or more 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′-fluorine modified nucleotides, 2′-deoxy modified nucleotides, and 2′-hydroxy modified nucleotides.

[0124] In some embodiments, the oligonucleotide is siRNA.

[0125] In some embodiments, the siRNA comprises a sense strand and an antisense strand that complement each other to form a double helix.

[0126] On the other hand, this application provides a pharmaceutical composition comprising the compounds, scaffolds, or conjugates of the preceding claims, and optionally a pharmaceutically acceptable carrier.

[0127] On the other hand, this application provides a kit comprising the compound, scaffold, or conjugate of the preceding claims.

[0128] Other aspects and advantages of this application will readily be apparent to those skilled in the art from the detailed description below. Only exemplary embodiments of this application are shown and described in the following detailed description. As will be appreciated by those skilled in the art, the content of this application enables them to make modifications to the disclosed specific embodiments without departing from the spirit and scope of the invention to which this application pertains. Accordingly, the descriptions in the accompanying drawings and specification of this application are merely exemplary and not restrictive. Attached Figure Description

[0129] The specific features of the invention involved in this application are shown in the appended claims. The features and advantages of the invention can be better understood by referring to the exemplary embodiments and drawings described in detail below. A brief description of the drawings is as follows:

[0130] Figure 1A shows the TTR-Q1 justice chain MS map described in this application;

[0131] Figure 1B shows the MS map of the TTR-Q1 antisense chain (which is also the TTR-Q2 antisense chain) described in this application;

[0132] Figure 1C shows the MS spectrum of the TTR-Q1 double strand described in this application;

[0133] Figure 1D shows the TTR-Q2 justice chain MS map described in this application;

[0134] Figure 1E shows the MS spectrum of the TTR-Q2 double strand described in this application;

[0135] Figure 2A shows the MS map of the ApoC3-Q1 justice chain described in this application;

[0136] Figure 2B shows the MS map of the ApoC3-Q1 antisense chain (which is also the ApoC3-Q2 antisense chain) described in this application;

[0137] Figure 2C shows the MS spectrum of the ApoC3-Q1 duplex described in this application;

[0138] Figure 2D shows the MS map of the ApoC3-Q2 justice chain described in this application;

[0139] Figure 2E shows the MS spectrum of the ApoC3-Q2 duplex described in this application;

[0140] Figure 3 shows the effect of the compound described in this application on inhibiting TTR;

[0141] Figure 4 shows the effect of the compound described in this application on inhibiting ApoC3. Detailed Implementation

[0142] The following specific embodiments illustrate the implementation of the invention. Those skilled in the art can easily understand other advantages and effects of the invention from the content disclosed in this specification.

[0143] Terminology Definition

[0144] The term "pharmaceutically acceptable" in this application means that the compound or composition is chemically and / or toxicologically compatible with other components constituting the formulation and / or with humans or mammals for the prevention or treatment of diseases or conditions.

[0145] The term "pharmaceutically acceptable salt" refers to a relatively non-toxic inorganic or organic acid addition salt of the compounds of this invention. For example, see SMBerge et al., "Pharmaceutical Salts," J. Pharm. Sci. 1977, 66, 1-19. Inorganic acids include, for example, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, or nitric acid; organic acids include, formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, hexanoic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)-benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, bamoic acid, pectinic acid, 3-phenylpropionic acid, picric acid, etc. Pteropenic acid, 2-hydroxyethanesulfonic acid, itaconic acid, aminosulfonic acid, trifluoromethanesulfonic acid, dodecyl sulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucoheponic acid, glycerophosphate, aspartic acid, sulfosalicylic acid, etc. For example, HCl (or hydrochloric acid), HBr (or hydrobromic acid solution), methanesulfonic acid, sulfuric acid, tartaric acid, or fumaric acid can be used to form pharmaceutically acceptable salts with the compounds shown in formula (I).

[0146] In this application, the compounds comprise their tautomers, meso compounds, racemates, enantiomers, and / or diastereomers. In this application, the term "diastereomer" generally refers to a stereoisomer having two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers can have different physical properties, such as melting point, boiling point, spectral properties, and reactivity. In this application, the terms "tautomer" or "tautomer form" are used interchangeably and generally refer to structural isomers of different energies that can be interconverted through a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via the rearrangement of some bonding electrons. In this application, the term "meta-form" generally refers to a molecule containing asymmetric atoms but possessing symmetry factors that result in zero total optical rotation within the molecule. The terms "racemic form" or "racemic mixture" refer to a composition consisting of two enantiomers in equimolar amounts.

[0147] In this application, the term "ligand" generally refers to any compound or molecule capable of covalently or otherwise chemically binding to a biologically active substance (such as an oligonucleotide). In some embodiments, the ligand is capable of interacting directly or indirectly with another compound, such as a receptor. The receptor interacting with the ligand may be present on the cell surface, or alternatively may be an intracellular and / or intercellular receptor. The interaction between the ligand and the receptor may result in a biochemical reaction, or may simply be a physical interaction or binding.

[0148] In this application, certain atoms of the compounds may appear in more than one isotopic form. For example, hydrogen may appear as protium (…). 1 H), deuterium ( 2 H) and tritium ( 3 Carbon exists in the form of H, and it may exist in three different isotopes (H). 12 C 13 C and 14 C) Naturally occurring. Examples of isotopes that may be incorporated into the compounds of this application include, but are not limited to, those that exist naturally. 15 N、 18 O、 17 O、 18 F, 32 P, 33 P, 129 I, 131 I, 123 I, 124I, 125 I, or similar isotopes. Therefore, the compounds of this application can be enriched in one or more of these isotopes relative to their natural abundance. As those skilled in the art will know, such isotope-enriched compounds can be used for a variety of purposes. For example, with heavy isotopes such as deuterium (I, or similar isotopes). 2 H) substitution may offer certain therapeutic advantages, possibly due to greater metabolic stability. For example, deuterium (H) 2 The natural abundance of deuterium (H) is approximately 0.015%. Therefore, there is approximately one deuterium atom for every 6500 hydrogen atoms in nature. Thus, the deuterium-containing compounds of this application have a deuterium abundance greater than 0.015% at one or more positions (as the case may be). Unless otherwise specified, the structures described in this application may also include compounds that differ only in the presence or absence of one or more isotopically enriched atoms. For example, compounds whose structures are identical to those of this application except that hydrogen atoms are replaced by deuterium or tritium, or carbon atoms are replaced by carbon-13 or carbon-14, are within the scope of this application.

[0149] In this application, the term "pharmaceutical composition" generally refers to a mixture containing one or more of the compounds described in this application or their physiologically / pharmaceutical-grade salts or prodrugs, along with other chemical components, such as physiologically / pharmaceutical-grade carriers and excipients. Pharmaceutical compositions can facilitate administration to an organism, promote the absorption of the active ingredient, and thereby exert its biological activity. Conventional methods for preparing pharmaceutical compositions are described in the art.

[0150] In this application, the term "alkyl" generally refers to a residue derived from an alkane by removing a hydrogen atom. Alkyl groups can be substituted or unsubstituted, substituted or non-substituted. The term "alkyl" generally refers to a saturated straight-chain or branched aliphatic hydrocarbon group having residues derived from the removal of hydrogen atoms from the same carbon atom or two different carbon atoms of the parent alkane, and can be a straight-chain or branched group containing 1 to 20 carbon atoms, for example, containing 1 to 12 carbon atoms, such as a chain alkyl containing 1 to 6 carbon atoms. Non-limiting examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, propyl, butyl, etc. Alkyl groups can be substituted or unsubstituted, alternative or non-substituted. For example, when substituted, the substituent can be replaced at any usable connection point. The substituent can be independently selected from one or more substituents chosen from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, and oxo, for example, it can be hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(=O)H, -CO2H, -C(=O)C(=O)H, -C(=O)CH2C(=O)H, -S(=O)H, -C(O)NH2, -SO2NH2, -OC(=O)H, -N(H)SO2H, or C 1-6 Aliphatic groups.

[0151] In this application, the term "alkylene" generally refers to a saturated straight-chain or branched aliphatic hydrocarbon group having two residues derived from the removal of two hydrogen atoms from the same carbon atom or two different carbon atoms of a parent alkane. It can be a straight-chain or branched group containing 1 to 20 carbon atoms. For example, the term "methylene" can refer to a residue derived from the removal of two hydrogen atoms from a 1-carbon group. The methylene group can be substituted or unsubstituted, substituted or non-substituted; for example, containing 1 to 12 carbon atoms, such as an alkylene group containing 1 to 6 carbon atoms. Non-limiting examples of alkylene groups include, but are not limited to, methylene (-CH2-), 1,1-ethylene (-CH(CH3)-), 1,2-ethylene (-CH2CH2-), 1,1-propylene (-CH(CH2CH3)-), 1,2-propylene (-CH2CH(CH3)-), 1,3-propylene (-CH2CH2CH2-), 1,4-butylene (-CH2CH2CH2CH2-), and 1,5-butylene (-CH2CH2CH2CH2CH2-). The alkylene group can be substituted or unsubstituted, alternative or non-alternative. For example, when substituted, the substituent can be substituted at any usable connection point. The substituent can be independently selected from one or more substituents chosen from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, and oxo, for example, it can be hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, or C1-6 aliphatic groups. The methylene or alkylene groups can be substituted or unsubstituted.

[0152] In this application, the term "alkenyl" generally refers to a straight-chain or branched hydrocarbon group containing one or more double bonds. Exemplary examples of alkenyl groups include allyl, homoallyl, vinyl, crotonyl, butenyl, pentenyl, and hexenyl. Exemplary examples of C2-6 chain alkenyl groups having more than one double bond include butadienyl, pentadienyl, hexadienyl, and hextrienyl, as well as their branched forms. The unsaturated bond (double bond) can be located at any position on the carbon chain. Alkenyl groups can be substituted or unsubstituted.

[0153] In this application, the term "alkenyl" generally refers to a residue derived from the removal of two hydrogen atoms from a carbon atom of an alkene. Examples include allylene, vinylene, butenylene, pentenylene, and hexenylene. Alkenyl groups can be substituted or unsubstituted.

[0154] In this application, the term "alkynyl" generally refers to an unsaturated straight-chain or branched alkynyl group, such as ethynyl, 1-propynyl, propynyl, butynyl, etc. The alkynyl group can be substituted or unsubstituted.

[0155] In this application, the term "ynynyl" generally refers to a residue derived from the removal of two hydrogen atoms from a carbon atom of an alkyne. For example, it can be ethynyl, propynyl, propynyl, butynyl, etc. The ynynyl group can be substituted or unsubstituted.

[0156] In this application, the term "aryl" generally refers to a residue derived from the removal of a hydrogen atom from an aromatic ring. The term "aromatic ring" can refer to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., a ring sharing adjacent carbon atom pairs) having a conjugated π-electron system, and can be 6- to 10-membered, such as benzene and naphthalene. The aromatic ring can be fused to a heteroaryl, heterocyclic, or cycloalkyl ring, wherein the ring attached to the parent structure is an aryl ring. The aryl group can be substituted or unsubstituted, and when substituted, the substituent can be one or more of the following groups, independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxy, heterocyclic alkoxy, cycloalkylthio, and heterocyclic alkylthio. The aryl group can be substituted or unsubstituted.

[0157] In this application, the term "arylene" generally refers to a residue derived by removing two hydrogen atoms from a carbon atom of an aromatic ring. For example, it could be phenylene or naphthylene. The arylene can be substituted or unsubstituted.

[0158] In this application, the term "heteroaryl" generally refers to a residue derived from the removal of a hydrogen atom from a carbon atom of a heteroaryl ring. The term "heteroaryl ring" refers to a heteroaryl system comprising 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms may be selected from the group consisting of oxygen, sulfur, and nitrogen. The heteroaryl group may be 5 to 10-membered, or 5- or 6-membered, such as furanyl, thiophene, pyridinyl, pyrroleyl, N-alkylpyrroleyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl ring may be fused to an aryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is the heteroaryl ring. The heteroaryl group can be optionally substituted or unsubstituted. When substituted, the substituent can be one or more of the following groups, independently selected from the group consisting of: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio. The heteroaryl group can be substituted or unsubstituted.

[0159] In this application, the term "heteroaryl" generally refers to a residue derived from the removal of two hydrogen atoms from a carbon atom of a heteroaryl ring. Examples include iminofuranyl, iminothienyl, iminopyridyl, iminopyrroleyl, iminopyridyl, iminopyrazinyl, iminopyrazolyl, iminotetrazoleyl, etc. Heteroaryl groups can be substituted or unsubstituted.

[0160] In this application, the term "alicyclic group" generally refers to a residue derived by removing a hydrogen atom from the same carbon atom or multiple different carbon atoms of an aliphatic ring. The term "cycloalkane" generally refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon having a carbocyclic ring containing 3 to 20 carbon atoms, or 3 to 12 carbon atoms, or 3 to 10 carbon atoms, or 3 to 8 carbon atoms. Non-limiting examples of alicyclic groups include cyclopropane, cyclobutane, cyclopentane, cyclopentenyl, cyclohexane, cyclohexenyl, cyclohexadienyl, cycloheptane, cycloheptanetrienyl, cyclooctane, etc.; polycyclic carbocyclic rings may include spirocyclic, fused-ring, and bridged-ring carbocyclic rings. Alicyclic groups can be substituted or unsubstituted. In this application, the term "carbocyclic group" generally refers to a residue derived by removing a hydrogen atom from a carbon atom of a carbocyclic ring. The term "carbocyclic ring" generally refers to saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbons containing 3 to 20 carbon atoms, 3 to 12 carbon atoms, 3 to 10 carbon atoms, or 3 to 8 carbon atoms. Non-limiting examples of monocyclic carbocyclic rings include cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexene, cyclohexadiene, cycloheptane, cycloheptanetriene, cyclooctane, etc.; polycyclic carbocyclic rings can include spirocyclic, fused, and bridged rings. The carbocyclic group can be substituted or unsubstituted. In some cases, alicyclic and carbocyclic rings can be used interchangeably.

[0161] In this application, the term "partially unsaturated" generally refers to a cyclic structure in which the cyclic molecules contain at least one double or triple bond. The term "partially unsaturated" encompasses cyclic structures with multiple unsaturations, but is not intended to include aromatic or heteroaromatic rings as defined in this application. The term "unsaturated" indicates that a portion has one or more degrees of unsaturation.

[0162] In this application, the term "alicyclic group" generally refers to a residue derived from the removal of two hydrogen atoms from a carbon atom of an alicyclic ring. For example, it can be cyclopropane, cyclobutane, cyclopentane, cyclopentenyl, cyclohexane, cyclohexenyl, cyclohexadienyl, cycloheptane, cycloheptanetrienyl, cyclooctyl, etc.; the polycyclic carbocyclic ring can include spirocyclic, fused-ring, and bridged-ring carbocyclic rings. The alicyclic group can be substituted or unsubstituted.

[0163] In this application, the term "aliphatic heterocyclic group" generally refers to a stable, non-aromatic 3-7 member monocyclic carbon ring structure, a fused 7-10 member bicyclic heterocyclic structure, or a bridged 6-10 member bicyclic heterocyclic structure. These cyclic structures can be saturated or partially saturated, and in addition to carbon atoms, they contain one or more heteroatoms selected from the group consisting of oxygen, sulfur, and nitrogen. For example, it may contain 1-4 heteroatoms as defined above. When used to refer to atoms in an aliphatic heterocyclic ring structure, the term "nitrogen" may include nitrogen that has undergone a substitution reaction. For example, an aliphatic heterocyclic group may include "heterocyclic alkyl group," which may refer to a stable, non-aromatic 3-7 member monocyclic alkyl structure, a fused 7-10 member bicyclic heterocyclic structure, or a bridged 6-10 member bicyclic heterocyclic structure. In addition to carbon atoms, these cyclic structures contain one or more heteroatoms selected from the group consisting of oxygen, sulfur, and nitrogen. For example, it may contain 1-4 heteroatoms as defined above. Heterocyclic alkyl groups can be substituted or unsubstituted. Alicyclic groups can be substituted or unsubstituted.

[0164] In this application, the term "allelic heterocyclic group" generally refers to a residue derived from the removal of two hydrogen atoms from a carbon atom of an alicyclic ring. Allelic heterocyclic groups can be substituted or unsubstituted.

[0165] In this application, the terms “optional” or “optionally” generally refer to an event or environment described subsequently that may but does not have to occur, and the description includes situations in which the event or environment occurs or does not occur. For example, “optionally alkyl-substituted heterocyclic group” means that an alkyl group may but does not have to be present, and the description can include cases where the heterocyclic group is substituted with an alkyl group and cases where the heterocyclic group is not substituted with an alkyl group.

[0166] In this application, the term "substituted" generally refers to one or more hydrogen atoms in a group, for example, up to five, or for example, one to three hydrogen atoms, independently substituted by the corresponding number of substituents. Substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without much effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when bonded to a carbon atom with an unsaturated bond (such as an alkene).

[0167] In this application, as those skilled in the art will know, terms such as "alkyl," "alkenyl," and "cycloalkyl" may be preceded by an identifier indicating the number of atoms present in the group under specific conditions, for example, C1-C4 alkyl, C3-C7 cycloalkoxy, C1-C4 alkylcarbonylamino, etc., and the subscript number following "C" indicates the number of carbon atoms present in the group. For example, C3 alkyl refers to an alkyl group having three carbon atoms (e.g., n-propyl, isopropyl); C 1-10In this context, the members of the group can have any number of carbon atoms falling within the range of 1-10.

[0168] One or more hydrogen atoms in a group, for example, up to five, or for example, one to three hydrogen atoms, are independently replaced by the corresponding number of substituents. The substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without much effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when bonded to a carbon atom with an unsaturated bond (such as an alkene).

[0169] In this application, the term "compound" generally refers to a substance having two or more different elements. For example, the compound in this application can be an organic compound, a compound with a molecular weight of less than 500, less than 1000, more than 1000, or more than 10,000 or 100,000. In this application, a compound can also refer to a compound linked by chemical bonds. For example, it can be a compound in which one or more molecules with a molecular weight of less than 1000 are linked by chemical bonds to a biological macromolecule, such as a polysaccharide, protein, nucleic acid, polypeptide, etc. For example, the compound in this application can include a compound in which a protein is linked to one or more molecules with a molecular weight of less than 1000, a compound in which a protein is linked to one or more molecules with a molecular weight of less than 10,000, or a compound in which a protein is linked to one or more molecules with a molecular weight of less than 100,000.

[0170] In this application, the term "comprising" generally means including the explicitly specified features, but does not exclude other elements. The terms "above" and "below" generally refer to situations that include the stated number.

[0171] In this application, the term "about" generally refers to a variation within a range of 0.5% to 10% above or below a specified value, such as a variation within a range of 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% above or below a specified value.

[0172] In this application, Ac represents acetyl group.

[0173] When connecting atoms, Represents a chemical bond. This indicates that other groups are connected here.

[0174] As used in this article, the term "protecting group" can refer to any atom or group of atoms introduced into a molecule to prevent existing groups in the molecule from undergoing undesirable chemical reactions, which can be removed to leave unprotected groups.

[0175] The hydroxyl protecting group can be, for example, the hydroxyl protecting group commonly used to protect the ribose structure in the synthesis of RNA or its derivatives, see also Green et al., Protective Groups in Organic Synthesis, 3rd Edition, 1999, John Protecting groups described in the Wiley & Sons, Inc. literature include, for example: acetyl, phenoxyacetyl, neopentanoyl, benzyl, 4-methoxybenzyl, benzoyl, triphenylmethyl, 4,4'-dimethoxytriphenylmethyl (DMTr), monomethoxytriphenylmethyl (MMTr), 9-phenyl-xanthen-9-yl, 9-(p-tolyl)-xanthen-9-yl, trimethylsilyl, tert-butyldimethylsilyl (TBDMS), cyanomethoxymethyl, 2-(cyanoethoxy)ethyl, cyanoethoxymethyl, etc., preferably 4,4'-dimethoxytriphenylmethyl (DMTr, 4,4'-dimethoxytrityl).

[0176] The amino protecting group may be, but is not limited to, 9-fluorenylmethoxycarbonyl (Fmoc), tert-butoxycarbonyl (BOC), benzyloxycarbonyl (Cbz), optionally substituted acyl, trifluoroacetyl (TFA), benzyl, triphenylmethyl (Tr), 4,4'-dimethoxytriphenylmethyl (DMTr) or toluenesulfonyl (Ts), preferably 9-fluorenylmethoxycarbonyl (Fmoc).

[0177] The term "phosphate group" is a phosphoric acid-containing group, which in some cases can be negatively charged. The charge is evenly distributed between the two unlinked oxygen atoms. In this application, the phosphate group can be linked to multiple L atoms, or it can be linked to an L atom and an oligonucleotide. However, the phosphate group can be modified by replacing one of the oxygen atoms with different substituents. Examples of modified phosphate groups include thiophosphate, phosphoroselenate, boron phosphate, boron phosphate ester, hydrophosphonate, aminophosphate, alkyl or aryl phosphonate, and phosphate triester. In dithiophosphate, both unlinked oxygen atoms are replaced by sulfur. The oxygen in the phosphodiester bond can be replaced by any one of S, Se, B, C, H, N, or OR (R is alkyl or aryl). The phosphate linker can also be modified by replacing the linking oxygen atoms with nitrogen (bridged aminophosphate), sulfur (bridged thiophosphate), and carbon (bridged methylene phosphonate).

[0178] As used herein, the term "phosphorus-containing reactive group" refers to a phosphorus-containing group capable of reacting with a hydroxyl or amino group contained in another molecule, particularly in another nucleotide unit or another nucleotide analog, via a nucleophilic attack reaction. Typically, such a reaction produces an ester-type nucleoside bond linking the nucleotide unit or nucleotide analog unit to another nucleotide unit or nucleotide analog unit. These phosphorus-containing reactive groups are known in the art and contain phosphorus atoms in the PIII or PV valence states, including but not limited to phosphorus amides, H-phosphonates, phosphate triesters, and phosphorus-containing chiral auxiliaries. Examples of phosphorus-containing reactive groups include: 2-cyanoethoxy-N,N-diisopropylaminophospho, 2-propenoxy-N,N-diisopropylaminophospho, methoxy-N,N-diisopropylaminophospho, bis(diisopropylaminophospho), etc.

[0179] 2-Cyanethoxy-N,N-diisopropylaminophospho,

[0180] 2-Acryloxy-N,N-Diisopropylaminophospho,

[0181] Methoxy-N,N-diisopropylaminophosphine,

[0182] Bis(diisopropylamino)phospho, etc.

[0183] In one embodiment, the phosphorus-containing reactive group is...

[0184] Invention Details

[0185] 1 Connector Unit

[0186] This application constructs linker units (e.g., IL, IW) based on 3,4-position substituted pyrrole rings for linking nucleotide drugs (e.g., siRNA) and ligands (e.g., GalNAc).

[0187] In some embodiments, nucleotide drug delivery systems (e.g., IX) or oligonucleotide conjugates formed with this linker unit have liver-targeted delivery efficiency comparable to or better than existing structurally similar GalNAc compounds.

[0188] In some embodiments, nucleotide drug delivery systems (e.g., IX) or oligonucleotide conjugates formed with this linker unit have comparable or better long-lasting effects compared to existing structurally similar GalNAc compounds.

[0189] In some embodiments, nucleotide drug delivery systems (e.g., IX) or oligonucleotide conjugates formed with this linker unit require an effective dose that is comparable to or lower than that of existing structurally similar GalNAc compounds.

[0190] In some embodiments, nucleotide drug delivery systems (e.g., IX) or oligonucleotide conjugates formed from this linker unit are easier and more economical to synthesize than existing structurally similar GalNAc compounds, and are suitable for scale-up synthesis and production.

[0191] On the one hand, this application provides a connecting body unit L, the general structural formula of which is (IL):

[0192] On the other hand, the linker unit L can be further linked to W, and this disclosure provides compounds of formula (IW) or pharmaceutically acceptable salts thereof:

[0193] The formula IW, where L connects to W, can serve as an intermediate in the production of nucleotide drug delivery systems. For example, when X2 is an amino group, W is an amino protecting group, such as Fmoc. In some embodiments, W in formula IW can be H, in which case IL has the same general formula structure as IW.

[0194] On the other hand, the linker unit L can be further linked to X, and this disclosure provides compounds of formula (IX) or pharmaceutically acceptable salts thereof:

[0195] In some implementations, formula (IL) has the following structure:

[0196] W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl groups optionally substituted with one or more halogens, amino substituents, or amino protecting groups. The W in the molecular formula disclosed in this application is interchangeable; for a specific description of W, please refer to the relevant description of W in section 1.1 of this application.

[0197] X is a ligand or a derivative thereof. The term X in the molecular formula disclosed in this application is interchangeable; for a specific description of X, please refer to the relevant description of X in section 1.2 of this application.

[0198] Y includes one of hydrogen, a hydroxyl protecting group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group.

[0199] The phosphate group of Y may include -P(R) Y1 2. -P(OR) Y1 )(N(R Y1 )2), -P(=O)(OR Y1 )R Y1 -P(=S)(OR Y1 )R Y1 -P(=O)(SR) Y1 )R Y1 -P(=S)(SR) Y1 )R Y1 -P(=O)(OR) Y1 )2、-P(=S)(OR Y1 )2、-P(=O)(SR Y1 )2、-P(=S)(SR Y1 )2、-P(=O)(NHSO2CH3)R Y1 One of them, wherein each R Y1 Independently, it is hydrogen, oxygen, hydroxyl, or a C1-C6 alkyl group optionally substituted with one or more halogens or cyano groups. In some embodiments, when used to connect multiple L groups, Y can be a subunit, such as -P(=O)(OH)- or -P(=S)(OH)-.

[0200] In the molecular formula disclosed in this application, Y can be used interchangeably. For a specific description of Y, please refer to the relevant description of Y in section 1.4 of this application.

[0201] Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group.

[0202] The phosphate group of Z may include -P(R) Z1 2. -P(OR) Z1 )(N(R Z1 )2), -P(=O)(ORZ1 )R Z1 -P(=S)(OR Z1 )R Z1 -P(=O)(SR) Z1 )R Z1 -P(=S)(SR) Z1 )R Z1 -P(=O)(OR) Z1 )2、-P(=S)(OR z1 )2、-P(=O)(SR z1 )2、-P(=S)(SR z1 )2、-P(=O)(NHSO2CH3)R z1 One of them, wherein each R z1 Independently, it is hydrogen, oxygen, hydroxyl, or a C1-C6 alkyl group optionally substituted with one or more halogens or cyano groups. In some embodiments, when used to connect multiple L groups, Z can be a subunit, such as -P(=O)(OH)- or -P(=S)(OH)-.

[0203] In the molecular formula disclosed in this application, Z can be used interchangeably. For a specific description of Z, please refer to the relevant description of Z in section 1.5 of this application.

[0204] X1 is a branched chain. X2 is a nucleophilic or electrophilic group. X1 or X2 in the molecular formula disclosed in this application can be used interchangeably. For a specific description of Z, please refer to the relevant descriptions of X1 and X2 in section 1.3 of this application.

[0205] R1-R6 are arbitrary substituents. In the molecular formula disclosed in this application, R1-R6... 12 It's generally applicable; the specifics regarding R1-R... 12 For a description, please refer to 1.6R1-R of this application. 12 Related descriptions.

[0206] These substituents will be described in detail below.

[0207] 1.1 W

[0208] W includes one of the following groups: hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl or amino substituents optionally substituted with one or more halogens, and amino protecting groups.

[0209] In some embodiments, W may be selected from the group consisting of: hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl (such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, n-butyl, tert-butyl, tert-butyl, pentyl, or hexyl) optionally substituted with one or more halogens (such as F, Cl, Br, or I), amino protecting group, optionally substituted acyl group, and amino substituent.

[0210] In some embodiments, W can be R W1 , where R W1 C1-C is an optional substitute 30 Alkyl groups, wherein the substituents optionally included may be selected from the group consisting of: halogen, cyano, hydroxy, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C3 alkyl polyoxyethylene, or C1-C3 alkyl polyoxypropylene.

[0211] In some embodiments, W may be methyl, ethyl, or propyl.

[0212] In some embodiments, W can be an amino substituent, i.e., a group suitable for replacing hydrogen in an amino group.

[0213] In some embodiments, W may be an amino protecting group, including but not limited to 9-fluorenylmethoxycarbonyl (Fmoc), tert-butoxycarbonyl (BOC), benzyloxycarbonyl (Cbz), optionally substituted acyl, trifluoroacetyl (TFA), benzyl, triphenylmethyl (Tr), 4,4′-dimethoxytriphenylmethyl (DMTr) or toluenesulfonyl (Ts).

[0214] In some embodiments, W can be hydrogen.

[0215] In some embodiments, W is trifluoroacetyl (TFA).

[0216] In some embodiments, W is an amino substituent, i.e., a group suitable for replacing the hydrogen atom of the amino group, such as -C(=O)R. W1 -C(=O)NHR W1 -C(=S)R W1 or -C(=S)NHR W1 , where R W1 C1-C is an optional substitute 30 Alkyl, wherein the optionally substituted group may be selected from the group consisting of: halogen, cyano, hydroxy, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C3 alkyl polyoxyethylene, or C1-C3 alkyl polyoxypropylene.

[0217] In some embodiments, W can be -R W2 C(=O)R W1 -R W2C(=O)NHRw1、-R W2 C(=S)R W1 or -R W1 C(=S)NHRw1, where R W1 C1-C is an optional substitute 30 Alkyl, wherein the substituents optionally substituted may be selected from the group consisting of: halogen, cyano, hydroxy, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C3 alkyl polyoxyethylene, or C1-C3 alkyl polyoxypropylene. R W2 It is one of N, O, and S.

[0218] In some embodiments, compounds of formula (IW) or pharmaceutically acceptable salts thereof further have the following general formula:

[0219] In some embodiments, compounds of formula (IW) or pharmaceutically acceptable salts thereof further have the following general formula:

[0220] 1.2 X

[0221] Ligand X may be galactose, galactosamine, N-acetylgalactosamine, mannose, glucose, glucosamine, N-acetylglucosamine, fucose, or lactose. Ligand X may also be N-acetylgalactosamine, galactose, galactosamine, N-formyl-galactosamine, N-propionyl-galactosamine, N-butyryl-galactosamine, or N-isobutyryl-galactosamine, or derivatives thereof, wherein the acyl group is acetyl or benzoyl.

[0222] In some embodiments, ligand X is N-acetylgalactosamine with a hydroxyl group fully protected by an acyl group, or N-acetylgalactosamine (GalNAc), and further has -(CH2). r The C(=O)- structure has the following molecular formula:

[0223] Where r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5. In some embodiments, r is 1, 2, 3, 4, or 5. For example, r can be 4.

[0224] Furthermore, in some embodiments, formula (IX) further has the following general formula.

[0225] Where r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5. In some embodiments, r is 1, 2, 3, 4, or 5. For example, r can be 4.

[0226] In some embodiments, ligand X may be a lipid or a lipid-based molecule.

[0227] For example, ligand X can be an alkyl group, such as C5-C. 31 Alkyl groups, such as C 10 -C 18 Alkyl groups, such as C 14 Alkyl, C 15 Alkyl, C 16 Alkyl, C 17 Alkyl, C 18 alkyl.

[0228] For example, ligand X can be alkenyl, such as C5-C. 31 alkenyl groups, such as C 10 -C 18 alkenyl groups, such as C 14 alkenyl, C 15 alkenyl, C 16 alkenyl, C 17 alkenyl, C 18 Alkenyl group. In one embodiment, ligand X comprises at least one double bond.

[0229] For example, ligand X can be an alkynyl group, such as C5-C. 31 Alkyne groups, such as C 10 -C 18 Alkyne groups, such as C 14 alkynyl group, C 15 alkynyl group, C 16 alkynyl group, C 17 alkynyl group, C 18 Alkyne group. In one embodiment, ligand X comprises at least one triple bond. In another embodiment, ligand X comprises at least one double bond and at least one triple bond.

[0230] 1.3 X1 and X2

[0231] X1 can be a branch.

[0232] Branches can include direct bonds or atoms such as oxygen or sulfur, and units such as NR. X11C(O), C(O)NH, SO, SO2, SO2NH, or atomic chains, such as substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, arylalkyl, arylalenyl, arylynyl, heteroarylalkyl, heteroarylalenyl, heteroarylynyl, heterocyclic alkyl, heterocyclic alkenyl, heterocyclic alkynyl, aryl, heteroaryl, heterocyclic, cycloalkyl, cycloalkenyl, alkylarylalkyl, alkylarylalenyl, alkylarylynyl, alkenylarylalkyl, alkenylarylalenyl, alkenylarylynyl, alkynylarylalkyl, alkynylarylalenyl, alkynylarylynyl Alkyl heteroarylalkyl, alkyl heteroarylalyl, alkyl heteroarylynyl, alkenyl heteroarylalkyl, alkenyl heteroarylalyl, alkenyl heteroarylynyl, ynyl heteroarylalkyl, ynyl heteroarylalyl, ynyl heteroarylynyl, alkyl heterocyclic alkyl, alkyl heterocyclic alkenyl, alkyl heterocyclic ynyl, alkenyl heterocyclic alkenyl, alkenyl heterocyclic alkenyl, alkenyl heterocyclic alkyl, ynyl heterocyclic alkenyl, ynyl heterocyclic ynyl, alkyl aryl, alkenyl aryl, ynyl aryl, alkyl heteroaryl, alkenyl heteroaryl, alkyl heterocyclic aryl, alkyl aryl, alkenyl aryl, ynyl heterocyclic aryl, alkyl aryl, alkenyl heteroaryl, alkyl heteroaryl, alkenyl heteroaryl, ynyl heterocyclic aryl, wherein one or more methylene groups may be affected by O, S, S(O), SO2, N(R) X11 2. C(O), a cleavable linking group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, or a substituted or unsubstituted heterocyclic group can interrupt or terminate the process; where R X11 It is hydrogen, acyl, aliphatic, or substituted aliphatic.

[0233] In some embodiments, the linker is a branched linker. The branch point of the branched linker can be at least trivalent, but can be a tetravalent, pentavalent, or hexavalent atom, or a group exhibiting such multiple valences. In some embodiments, the branch point is -C, -CH, -C(CH2-)(CH2-)CH2-, -C(H)(CH2-)CH2-N, -N(Q)-C, -OC, -SC, -SS-C, -C(=O)N(Q)-C, -OC(=O)N(Q)-C, -N(Q)C(=O)-C, or -N(Q)C(=O)OC; wherein Q is independently hydrogen or optionally substituted alkyl group for each occurrence. In other embodiments, the branch point is glycerol or a glycerol derivative.

[0234] X1 includes -(CH2) n -、-(CH2) n NH-, -(CH2) n O-, -(CH2) n S-、-R X12 (CH2) n -、-R X12 (CH2) n NH-, -R X12 (CH2) nO- or -R X12 (CH2) n One of S-; where R X12 It is one of N, O, and S; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

[0235] In some implementations, X1 may include the following structure: -(CH2) n -、-(CH2) n NH-, -(CH2) n O-, -(CH2) n S-、-R X12 (CH2) n -、-R X12 (CH2) n N-、-R X12 (CH2) n O- or -R X12 (CH2) n S-, and optional substitutions of the above structures, wherein the optionally substituted group can be substituted by a substituent selected from the group consisting of: halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C3 alkyl polyoxyethylene, or C1-C3 alkyl polyoxypropylene; wherein R X12 It is one of N, O, and S. n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5. In some embodiments, n is 1, 2, 3, 4, or 5. For example, n can be 5.

[0236] For example, the structure of X1 can be as follows:

[0237] For a description of R7 and R8, see 1.6R1-R 12 .

[0238] In some implementations, X1 includes -((CH2) m1 O) m2 -,-((CH2) m1 O) m2 CH2-, -CH2((CH2) m1 O) m2 CH2-, where m1 is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5; m2 is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5. For example, m1 is 2 and m2 is 2.

[0239] In some embodiments, X1 has the structure -CH2OCH2CH2OCH2CH2- or -(CH2)5-.

[0240] In some implementations, X1 includes -((CH2) m1 O) m2 -,-((CH2) m1 O) m2 CH2-, -CH2((CH2) m1 O) m2 Optional substitutions for CH2-. Wherein m1 is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5; m2 is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5. For example, m1 is 2, and m2 is 2. The substituents optionally attached to the group may be substituted by substituents selected from the group consisting of: halogen, cyano, hydroxy, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C3 alkyl polyoxyethylene, or C1-C3 alkyl polyoxypropylene.

[0241] For example, the structure of X1 can also be as follows:

[0242] R9-R 12 For a description, see 1.6R1-R 12 .

[0243] In some embodiments, compounds of formula (IL), formula (IW), or formula (IX) or pharmaceutically acceptable salts thereof further have the following general formula:

[0244] X2 is an electrophilic or affinity group. In some embodiments, X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-.

[0245] Furthermore, compounds of formula (IW) or formula (IX) or their pharmaceutically acceptable salts further have the following general formula:

[0246] In the above chemical formula, n can be 5 and m2 can be 2.

[0247] 1.4 Y

[0248] The main function of Y is to connect multiple linker units L or oligonucleotides.

[0249] Y includes one of the following groups: hydrogen, C1-C6 alkyl groups optionally substituted with one or more halogens, phosphate groups, hydroxyl protecting groups, substituted acyl groups, and each R Y1 It is independently hydrogen, oxygen, hydroxyl, or a C1-C6 alkyl group optionally substituted with one or more halogens or cyano groups.

[0250] The phosphate group may include -P(R) Y1 2. -P(OR) Y1 )(N(R Y1 )2), -P(=O)(OR Y1 )R Y1 -P(=S)(OR Y1 )R Y1 -P(=O)(SR) Y1 )R Y1 -P(=S)(SR) Y1 )R Y1 -P(=O)(OR) Y1 )2、-P(=S)(OR Y1 )2、-P(=O)(SR Y1 )2、-P(=S)(SR Y1 )2、-P(=O)(NHSO2CH3)R Y1 One of them, wherein each R Y1 It is independently hydrogen, oxygen, hydroxyl, or a C1-C6 alkyl group optionally substituted with one or more halogens or cyano groups.

[0251] In some implementations, Y can be hydrogen.

[0252] In some embodiments, Y can be a C1-C6 alkyl group (such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, tert-butyl, pentyl, or hexyl) optionally substituted with one or more halogens (such as F, Cl, Br, or I).

[0253] In some embodiments, Y can be a C1-C6 alkyl group (such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, n-butyl, tert-butyl, tert-butyl, pentyl, or hexyl).

[0254] In some embodiments, Y can be methyl, ethyl, or propyl.

[0255] In some implementations, Y can be -P(R) Y1 2. -P(OR) Y1 )(N(R Y1 )2), -P(=O)(OR Y1 )R Y1 -P(=S)(ORY1 )R Y1 -P(=O)(SR) Y1 )R Y1 -P(=S)(SR) Y1 )R Y1 -P(=O)(OR) Y1 )2、-P(=S)(OR Y1 )2、-P(=O)(SR Y1 )2、-P(=S)(SR Y1 )2.

[0256] In some implementations, at least one R Y1 It is hydrogen. In some implementations, each R Y1 It is hydrogen.

[0257] In some implementations, at least one R Y1 It is a C1-C6 alkyl group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, tert-butyl, pentyl, or hexyl) optionally substituted with one or more halogens (e.g., F, Cl, Br, or I) or cyano groups.

[0258] In some implementations, each R Y1 It is a C1-C6 alkyl group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, n-butyl, tert-butyl, tert-butyl, pentyl, or hexyl) optionally substituted with one or more halogens (e.g., F, Cl, Br, or I) or cyano groups. In some embodiments, at least one R Y1 It is hydrogen, at least one R Y1 It is a C1-C6 alkyl group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, tert-butyl, pentyl, or hexyl) optionally substituted with one or more halogens or cyano groups.

[0259] In some implementations, Y can be -P(R) Y1 )2, for example, when R Y1 When it is hydrogen, Y is -PH2.

[0260] In some implementations, Y can be -P(OR) Y1 )(N(R Y1 )2), for example, when R Y1 When it is hydrogen, Y is -P(OH)(NH2).

[0261] In some embodiments, Y may be -P(O(C1-C6 alkyl))(N(C1-C6 alkyl)2), wherein the C1-C6 alkyl group is optionally substituted with one or more halogens or cyano groups.

[0262] In some implementations, Y can be -P (=O)(OR)Y1 )R Y1 .

[0263] In some embodiments, Y can be -P(=O)(OH)(C1-C6 alkyl), wherein the C1-C6 alkyl group is optionally substituted with one or more halogens or cyano groups.

[0264] In some implementations, Y can be -P (=S)(OR) Y1 )R Y1 .

[0265] In some embodiments, Y can be -P(=S)(OH)(C1-C6 alkyl), wherein the C1-C6 alkyl group is optionally substituted with one or more halogens or cyano groups.

[0266] In some implementations, Y is -P (=O)(SR) Y1 )R Y1 .

[0267] In some embodiments, Y is -P(=O)(SH)(C1-C6 alkyl), wherein the C1-C6 alkyl group is optionally substituted with one or more halogens or cyano groups.

[0268] In some implementations, Y is -P(=S)(SR) Y1 )R Y1 .

[0269] In some embodiments, Y is -P(=S)(SH)(C1-C6 alkyl), wherein the C1-C6 alkyl group is optionally substituted with one or more halogens or cyano groups.

[0270] In some implementations, Y is -P (=O)(OR) Y1 2. For example, when R Y1 When it is hydrogen, Y is -P(=O)(OH)2 or -P(=O)(OH)-.

[0271] In some implementations, Y is -P(=S)(OR) Y1 2. For example, when R Y1 When it is hydrogen, Y is -P(=S)(OH)2 or -P(=S)(OH)-.

[0272] In some implementations, Y is -P (=O)(SR) Y1 )2, for example, when R Y1 When it is hydrogen, Y is -P(=O)(SH)2.

[0273] In some implementations, Y is -P(=S)(SR) Y1 )2, for example, when R Y1When the hydrogen is used, in some embodiments, Y is -P(=S)(SH)2.

[0274] In some implementations, Y is -P(=O)(OH)2, or -P(=S)(OH)2, or -P(=S)(SH)(OH), or -P(=O)(OH)-, or -P(=S)(OH)-, or -P(=S)(SH)-, or -P(=O)(NHSO2CH3)(OH), or -P(=O)(NHSO2CH3)-, or -P(=S)(NHSO2CH3)(OH), or -P(=S)(NHSO2CH3)-.

[0275] In some embodiments, Y is a hydroxyl protecting group (e.g., silyl, Tr, DMTr, acyl, or benzyl). For example, Y can be silyl (e.g., trimethylsilyl, triethylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, or triisopropylsilyl), Y can be triphenylmethyl (Tr) or 4,4′-dimethoxytriphenylmethyl (DMTr), Y can also be an optionally substituted acyl (e.g., an optionally substituted acetyl) or benzyl.

[0276] In some embodiments, the compound of formula (IL), formula (IW) or formula (IX) or a pharmaceutically acceptable salt thereof further has one of the following structures:

[0277] 1.5 Z

[0278] The main function of Z is to connect multiple linker units L or oligonucleotides.

[0279] Z includes one of the following groups: hydrogen, C1-C6 alkyl groups optionally substituted with one or more halogens, phosphate groups, phosphorus-containing reactive groups, hydroxyl protecting groups, substituted acyl groups, and each R Z1 It is independently hydrogen, oxygen, hydroxyl, or a C1-C6 alkyl group optionally substituted with one or more halogens or cyano groups.

[0280] The phosphate group of Z may include -P(R) Z1 2. -P(OR) Z1 )(N(R Z1 )2), -P(=O)(OR Z1 )R Z1 -P(=S)(OR Z1 )R Z1 -P(=O)(SR) Z1 )R Z1 -P(=S)(SR) Z1 )R Z1-P(=O)(OR) Z1 )2、-P(=S)(OR z1 )2、-P(=O)(SR z1 )2、-P(=S)(SR z1 )2、-P(=O)(NHSO2CH3)R z1 One of them, wherein each R z1 Independently, Z can be hydrogen, oxygen, hydroxyl, or a C1-C6 alkyl group optionally substituted with one or more halogens or cyano groups. In some embodiments, when used to connect multiple L groups, Z can be a subunit, such as -P(=O)(OH)-, -P(=S)(SH)-, or -P(=S)(OH)-. In some embodiments, Z can be hydrogen.

[0281] In some embodiments, Z can be a C1-C6 alkyl group (such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, tert-butyl, pentyl, or hexyl) optionally substituted with one or more halogens (such as F, Cl, Br, or I).

[0282] In some embodiments, Z can be a C1-C6 alkyl group (such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, n-butyl, tert-butyl, tert-butyl, pentyl, or hexyl).

[0283] In some embodiments, Z can be methyl, ethyl, or propyl.

[0284] In some implementations, Z can be -P(R) Z1 2. -P(OR) Z1 )(N(R Z1 )2), -P(=O)(OR Z1 )RZ、-P(=S)(OR Z1 )R Z1 -P(=O)(SR) Z1 )R Z1 -P(=S)(SR) Z1 )R Z1 -P(=O)(OR) Z1 )2、-P(=S)(OR Z1 )2、-P(=O)(SR Z1 )2、-P(=S)(SR Z1 )2.

[0285] In some implementations, at least one R Z1 It is hydrogen. In some implementations, each R Z1 It is hydrogen.

[0286] In some implementations, at least one R Z1It is a C1-C6 alkyl group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, tert-butyl, pentyl, or hexyl) optionally substituted with one or more halogens (e.g., F, Cl, Br, or I) or cyano groups. In some embodiments, each R Z1 It is a C1-C6 alkyl group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, n-butyl, tert-butyl, tert-butyl, pentyl, or hexyl) optionally substituted with one or more halogens (e.g., F, Cl, Br, or I) or cyano groups. In some embodiments, at least one R Z1 It is hydrogen, at least one R Z1 It is a C1-C6 alkyl group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, tert-butyl, pentyl, or hexyl) optionally substituted with one or more halogens or cyano groups.

[0287] In some implementations, Z can be -P(R) Z1 )2, for example, when R Z1 When it is hydrogen, Z is -PH2.

[0288] In some implementations, Z can be -P(OR) Z1 )(N(R Z1 )2), for example, when R Z1 When it is hydrogen, Z is -P(OH)(NH2).

[0289] In some embodiments, Z can be -P(O(C1-C6 alkyl))(N(C1-C6 alkyl)2), wherein the C1-C6 alkyl group is optionally substituted with one or more halogens or cyano groups.

[0290] In some implementations, Z can be -P (=O)(OR) Z1 )R Z1 .

[0291] In some embodiments, Z can be -P(=O)(OH)(C1-C6 alkyl), wherein the C1-C6 alkyl group is optionally substituted with one or more halogens or cyano groups.

[0292] In some embodiments, Z can be -P(=S)(OR) Z1 )R Z1 .

[0293] In some embodiments, Z can be -P(=S)(OH)(C1-C6 alkyl), wherein the C1-C6 alkyl group is optionally substituted with one or more halogens or cyano groups.

[0294] In some implementations, Z is -P (=O)(SR) Z1 )R Z1 .

[0295] In some embodiments, Z is -P(=O)(SH)(C1-C6 alkyl), wherein the C1-C6 alkyl group is optionally substituted with one or more halogens or cyano groups.

[0296] In some implementations, Z is -P(=S)(SR) Z1 )R Z1 .

[0297] In some embodiments, Z is -P(=S)(SH)(C1-C6 alkyl), wherein the C1-C6 alkyl group is optionally substituted with one or more halogens or cyano groups.

[0298] In some implementations, Z is -P (=O)(OR) Z1 )2.

[0299] In some embodiments, Z is -P(=S)(OR) Z1 2. For example, when R Z1 When it is hydrogen, Z is -P(=S)(OH)2.

[0300] In some embodiments, Z is -P(=O)(SR) Z1 )2, for example, when R Z1 When it is hydrogen, Z is -P(=O)(SH)2.

[0301] In some implementations, Z is -P(=S)(SR) Z1 )2, for example, when R Z1 When Z is hydrogen, in some embodiments, Z is -P(=S)(SH)2.

[0302] In some embodiments, Z is -P(=O)(OH)2, or -P(=S)(OH)2, or -P(=S)(SH)(OH), or -P(=O)(OH)-, or -P(=S)(OH)-, or -P(=S)(SH)-, or -P(=O)(NHSO2CH3)(OH), or -P(=O)(NHSO2CH3)-, or -P(=S)(NHSO2CH3)(OH), or -P(=S)(NHSO2CH3)-.

[0303] In some embodiments, Z can be a hydroxyl protecting group (e.g., silyl, Tr, DMTr, acyl, or benzyl). For example, Z can be silyl (e.g., trimethylsilyl, triethylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, or triisopropylsilyl), Z can be triphenylmethyl (Tr) or 4,4′-dimethoxytriphenylmethyl (DMTr), Z can also be an optionally substituted acyl (e.g., an optionally substituted acetyl) or benzyl.

[0304] In one embodiment, Z can be a hydroxyl protecting group or a phosphorus-containing reactive group. The hydroxyl protecting group can be, for example, an acyl group (e.g., acetyl, phenoxyacetyl, 4-isopropylphenoxyacetyl), silyl, trimethyl, monomethoxytrimethyl (MMTr), or 4,4'-dimethoxytrimethyl (DMTr).

[0305] Phosphorus-containing reactive groups can be, for example:

[0306] In some embodiments, compounds such as those of formula (I) or pharmaceutically acceptable salts thereof further have the following general formula:

[0307] 1.6 R1-R 12

[0308] R1-R 12 It can be arbitrarily selected from the following group: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-6 alkyl, optional terminating C 1-6 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-6 alkyl carbonyl, C with optional substituent 1-6 Cycloalkyl carbonyl, optionally substituted heterocycloalkyl carbonyl, optionally substituted heterocycloalkylalkyl carbonyl, optionally substituted aryl carbonyl, or optionally substituted heteroaryl carbonyl. The groups with the optional substituted groups may be substituted by substituents selected from the group consisting of: halogen, cyano, hydroxy, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or C1-C6 alkyl polyoxypropylene.

[0309] In some embodiments, compounds such as those of formula (I) or pharmaceutically acceptable salts thereof further have the following general formula:

[0310] In formulas (IHW), (IHX), (I-OMe1-X), (I-Ome2-X), (I-Ome3-X), (I-OMe1-W), (I-Ome2-W), (I-Ome3-W), (I-F1-X), (I-F2-X), (I-F3-X), (I-F1-W), (I-F2-W), and (I-F3-W), R5 and / or R6 can be hydrogen. The substituents -OMe and -F can be replaced with other substituents, such as hydrogen, other halogens, hydroxyl groups, ethoxy groups, alkylamino groups, cycloalkylamino groups, heterocyclic groups, aryl groups, heteroaryl groups, and C6 groups. 1-3 Alkyl polyoxyethylene, C with optional substituents 1-6 alkyl, optional substituent C 1-6 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-6 alkyl carbonyl, C with optional substituent 1-6 Cycloalkyl carbonyl, optionally substituted heterocycloalkyl carbonyl, optionally substituted heterocycloalkylalkyl carbonyl, optionally substituted aryl carbonyl, or optionally substituted heteroaryl carbonyl. The groups with the optional substituted groups may be substituted by substituents selected from the group consisting of: halogen, cyano, hydroxy, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or C1-C6 alkyl polyoxypropylene.

[0311] 1.7 Support Structure

[0312] This application further includes a scaffold structure based on formula (IL), formula (IW), or formula (IX), which is linked by phosphate groups or thiophosphate groups of Y or Z, and multiple linker units L are connected by phosphate ester bonds or thiophosphate ester bonds to form a scaffold structure having multiple linker units L. This scaffold structure can be further linked with oligonucleotides to form a nucleotide drug delivery system.

[0313] The support structure may include one of the following:

[0314] Where q is an integer from 1 to 3, for example, q can be 1, 2, or 3.

[0315] Based on the compounds of formula (IW) and formula (IX), this application further provides a scaffold structure with the following molecular formula:

[0316] In some embodiments, the scaffold structure formed according to formula (IW) or such formula (IX) further has the following general formula:

[0317] The wavy line for O indicates that oxygen can be linked to hydrogen or oligonucleotides, while the wavy line for P indicates that it can be linked to a hydroxyl group, or to an oligonucleotide, or to the O group of an oligonucleotide.

[0318] In some embodiments, the scaffold structure formed according to formula (IW) or such formula (IX) further has the following general formula:

[0319] n is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5 (e.g., 1, 2, 3, 4, or 5), m2 is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5 (e.g., 1, 2, 3, 4, or 5), r is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5 (e.g., 1, 2, 3, 4, or 5), and RA is an oligonucleotide. In some embodiments, n is 5 and m2 is 2.

[0320] In some embodiments, the scaffold structure formed according to formula (IW) or such formula (IX) further has the following general formula:

[0321] n is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5 (e.g., 1, 2, 3, 4, or 5), and m2 is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5 (e.g., 1, 2, 3, 4, or 5). In some embodiments, n is 5 and m2 is 2.

[0322] 1.8 Other Exemplary Compounds

[0323] Based on the 3,4-substitution of the pyrrole ring, the 2 and 5 positions of the pyrrole ring can be arbitrarily substituted with -MeO, -F. For example, compounds of formula (IW) or formula (IX) further have the following general formula:

[0324] In some embodiments, the compound of formula (IW) or a pharmaceutically acceptable salt thereof further has one of the following structures:

[0325] Where r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5. In some embodiments, r is 1, 2, 3, 4, or 5. For example, r can be 4.

[0326] m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5. For example, m2 can be 1, 2, 3, 4, or 5. For example, m2 can be 2.

[0327] n can be 1, 2, 3, 4, or 5. For example, n can be 5.

[0328] In some embodiments, compounds of formula (IW) or formula (IX) further have the following general formula:

[0329] Where r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5. In some embodiments, r is 1, 2, 3, 4, or 5. For example, r can be 4.

[0330] m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5. For example, m2 is 1, 2, 3, 4 or 5.

[0331] n can be 1, 2, 3, 4, or 5. For example, n can be 5.

[0332] 2. Conjugates

[0333] On the other hand, this application further includes a conjugate having the following general structural formula:

[0334] in,

[0335] L refers to the aforementioned connecting body unit L;

[0336] X is the aforementioned ligand or its derivative;

[0337] RA stands for oligonucleotide;

[0338] p is 0-10, preferably 1-3, and more preferably 2.

[0339] In some embodiments, p can be 0, 1, 2, or 3.

[0340] Furthermore, the general structural formula of the conjugate is as follows:

[0341] RA stands for oligonucleotide.

[0342] Furthermore, the structural formula of the conjugate includes one of the following:

[0343] In some embodiments, the conjugate structure included in this application further includes one of the following:

[0344] n is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5. In some embodiments, n is 5; m2 is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5. In some embodiments, m2 is 2; RA is an oligonucleotide.

[0345] In some embodiments, the conjugate structure included in this application further includes one of the following:

[0346] n is an integer from 0 to 20, preferably 1 to 10, more preferably 1 to 5, and in some embodiments, n is 5;

[0347] m2 is an integer from 0 to 20, preferably from 1 to 10, more preferably from 1 to 5, and in some embodiments, m2 is 2;

[0348] RA stands for oligonucleotide.

[0349] The oligonucleotides in this application include single-stranded oligonucleotides (e.g., antisense nucleotides, or ASO) and double-stranded oligonucleotides (e.g., small interfering nucleotides, or siRNA). In a preferred embodiment, the oligonucleotide contains 7-30 nucleotides. Conjugates prepared from such oligonucleotides will have greater therapeutic value.

[0350] In one embodiment, the oligonucleotide is selected from small interfering nucleotides, DNA, microRNA (miRNA), small activating RNA (saRNA), small guide RNA (sgRNA), transfer RNA (tRNA), antisense nucleotides, or aptamers, with the oligonucleotide preferably being an antisense nucleotide or a small interfering nucleotide.

[0351] The nucleic acids of this invention can be single-stranded DNA or RNA, double-stranded DNA or RNA, or DNA-RNA hybrids. Examples of double-stranded DNA include structural genes, genes containing control and termination regions, and DNA from self-replication systems such as viruses or plasmids. Examples of double-stranded RNA include siRNA and other RNA interference agents. Single-stranded nucleic acids include, for example, antisense oligonucleotides, ribozymes, microRNAs, and oligonucleotides that form triple helices.

[0352] The nucleic acids of the present invention can have various lengths, which generally depend on the specific form of the nucleic acid. For example, in specific embodiments, the length of a plasmid or gene can range from about 1,000 to 100,000 nucleotide residues. In specific embodiments, the length of an oligonucleotide can range from about 10 to 100 nucleotides. In various related embodiments, the lengths of single-stranded, double-stranded, and triple-stranded oligonucleotides can range from about 10 to about 50 nucleotides, about 20 to about 50 nucleotides, about 15 to about 30 nucleotides, and about 20 to about 30 nucleotides.

[0353] In specific embodiments, the oligonucleotide (or its chain) of the present invention specifically hybridizes to or is complementary to the target polynucleotide. "Specifically hybridizable" and "complementary" are terms used to indicate sufficient complementarity to produce a stable and specific binding between the DNA or RNA target and the oligonucleotide. It should be understood that the oligonucleotide does not need to be 100% complementary to its target nucleic acid sequence for specific hybridization. The oligonucleotide is specifically hybridizable and has sufficient complementarity to avoid non-specific binding of the oligonucleotide to non-target sequences under conditions of desired specific binding, namely, physiological conditions in in vivo assays or therapeutic treatments, or conditions under which in vitro assays are performed. Therefore, in other embodiments, the oligonucleotide includes substitutions of 1, 2, or 3 bases compared to the region of the gene or mRNA sequence to which it targets or specifically hybridizes.

[0354] In specific embodiments, the compounds of the present invention are associated with RNA interference (RNAi) molecules. RNA interference methods using RNAi molecules can be used to disrupt the expression of genes or polynucleotides of interest. These RNAi molecules are also referred to as iRNA agents and are described below.

[0355] iRNA agents should include regions that are sufficiently homologous to the target gene and of sufficient length in terms of nucleotides to allow the iRNA agent or fragments thereof to mediate downregulation of the target gene. (For ease of interpretation, the terms nucleotide or ribonucleotide are sometimes used herein to refer to one or more monomeric subunits of the RNA agent. It will be understood herein that the use of the terms “ribonucleotide” or “nucleotide” may also refer to the modified nucleotide or the substitution portion at one or more positions in the case of modified RNA or nucleotide substitutions.) Thus, an iRNA agent is or includes a region that is at least partially complementary to the target RNA and, in one embodiment, completely complementary. Perfect complementarity between the iRNA agent and the target is not necessary, but conformity must be sufficient to allow the iRNA agent or its cleavage product to guide sequence-specific silencing, for example, through RNAi cleavage of the target RNA such as mRNA. The degree of complementarity or homology with the target strand is most critical in the antisense strand. When perfect complementarity (especially in the antisense strand) is generally desired, an embodiment may include (especially in the antisense strand) one or more, or for example, six, five, four, three, two, or fewer mismatches (relative to the target RNA). The mismatches, particularly those in the antisense strand, are most tolerant in the terminal regions and, if present, can be in the terminal regions or in regions of 6, 5, 4, or 3 nucleotides from, for example, the 5' and / or 3' ends. The sense strand only needs to be sufficiently complementary to the antisense strand to maintain the overall double-stranded nature of the molecule.

[0356] As discussed elsewhere in this document, and in material incorporated herein by reference in its entirety, iRNA agents are typically modified or include nucleoside substitutes. Single-stranded regions of iRNA agents are typically modified or include nucleoside substitutes, such as one or more unpaired regions of a hairpin structure, or regions connecting two complementary regions that may have modifications or nucleoside substitutes. Modifications that stabilize one or more 3' or 5' ends of iRNA agents, for example, targeting exonucleases, or that facilitate the entry of antisense siRNA agents into RISC are also envisioned. Modifications may include C3 (or C6, C7, C12) amino linkers, thiol linkers, carboxyl linkers, non-nucleotide spacer groups (C3, C6, C9, C12, debased, triethylene glycol, hexaethylene glycol), and specific biotinylate or luciferin reagents, wherein the luciferin reagent is phosphorousamide and has another hydroxyl group protecting DMT, allowing multiple coupling during RNA synthesis.

[0357] iRNA agents comprise: molecules long enough to trigger an interferon response and enter the RISC (RNAi-induced silencing complex) (which can be cleaved by Dicer (Bemstein et al. 2001. Nature, 409: 363-366); and molecules short enough that they do not trigger an interferon response (which can also be cleaved by Dicer and / or enter the RISC), such as molecules having a size that allows entry into the RISC, such as molecules similar to Dicer cleavage products. Molecules short enough that they do not trigger an interferon response are referred to herein as siRNA agents or shorter iRNA agents. As used herein, “siRNA agent or shorter iRNA agent” means an iRNA agent, such as a double-stranded or single-stranded RNA agent that is short enough that it does not induce a harmful interferon response in human cells, for example, having a double-stranded region of fewer than 60, 50, 40, or 30 nucleotide pairs. siRNA agents or their cleavage products can downregulate target genes by, for example, inducing RNAi with respect to target RNA, wherein the target may include endogenous or pathogenic target RNA.

[0358] The length of each strand of an siRNA agent can be equal to or less than 30, 25, 24, 23, 22, 21, or 20 nucleotides. The length of the strand can be at least 19 nucleotides. For example, the length of each strand can be between 21 and 25 nucleotides. The siRNA agent can have a double-stranded region of 17, 18, 19, 29, 21, 22, 23, 24, or 25 nucleotide pairs, and one or more overhangs, or one or two 3' overhangs of 2-3 nucleotides.

[0359] In addition to homology with target RNA and the ability to downregulate target genes, iRNA agents may possess one or more of the following properties:

[0360] The single-stranded iRNA agent can be long enough for it to enter RISC and participate in RISC-mediated cleavage of the target mRNA. The length of the single-stranded iRNA agent is at least 14 nucleotides, and in other embodiments at least 15, 20, 25, 29, 35, 40, or 50 nucleotides. In some embodiments, its length is less than 200, 100, or 60 nucleotides.

[0361] The hairpin iRNA agent will have a double-stranded region of 17, 18, 19, 29, 21, 22, 23, 24, or 25 nucleotide pairs. The length of the double-stranded region will be equal to or less than 200, 100, or 50 nucleotide pairs. In some embodiments, the length of the double-stranded region ranges from 15-30, 17-23, 19-23, and 19-21 nucleotide pairs. The hairpin structure is at 3' in one embodiment, and in some embodiments may have a single-stranded overhang or a terminal unpaired region on the antisense side of the hairpin structure. In one embodiment, the overhang is 2-3 nucleotides long.

[0362] The term "double-stranded (ds)iRNA agent" as used in this article refers to an iRNA agent comprising more than one strand, and in some cases two strands, in which regions of interstrand hybridization can form a double-stranded structure.

[0363] The antisense strand of a double-stranded iRNA agent can be equal to or at least 14, 15, 16, 17, 18, 19, 25, 29, 40, or 60 nucleotides in length. Its length can be equal to or less than 200, 100, or 50 nucleotides. The length range can be 17 to 25, 19 to 23, and 19 to 21 nucleotides.

[0364] The sense strand of a double-stranded iRNA agent can be equal to or at least 14, 15, 16, 17, 18, 19, 25, 29, 40, or 60 nucleotides in length. Its length can be equal to or less than 200, 100, or 50 nucleotides. The length range can be 17 to 25, 19 to 23, and 19 to 21 nucleotides.

[0365] The length of the double-stranded portion of a double-stranded iRNA agent can be equal to or at least 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 29, 40, or 60 nucleotide pairs. Its length can be equal to or less than 200, 100, or 50 nucleotide pairs. The length range can be 15-30, 17-23, 19-23, and 19-21 nucleotide pairs.

[0366] It is desirable to modify one or both of the antisense and sense strands of a double-stranded iRNA agent. In some cases, they will have the same or similar modifications, but in others, the sense and antisense strands will have different modifications, for example, in some cases it is desirable to modify only the sense strand. It may be desirable to modify only the sense strand to, for example, render it inactive, for example, the sense strand can be modified to render it inactive and prevent the formation of active siRNA / protein or RISC. This can be achieved by modifications that prevent 5'-phosphorylation of the sense strand. Other modifications that prevent phosphorylation can also be used, for example, simply replacing 5'-OH with H instead of O-Me. Alternatively, a large group can be added to the 5'-phosphate to convert it into a phosphodiester bond, although this may be less desirable because phosphodiesterases can cleave this bond and release the 5' end of the functional siRNA. Antisense strand modifications include 5' phosphorylation and any other 5' modifications discussed herein, particularly the 5' modifications discussed in the single-stranded iRNA molecule section above.

[0367] The sense and antisense strands can be selected such that the ds iRNA agent includes single-stranded or unpaired regions at one or both ends of the molecule. Therefore, the ds iRNA agent may contain a sense and antisense strand, which are paired to contain overhangs, such as one or two 5' or 3' overhangs, or 3' overhangs of 2-3 nucleotides. Many embodiments will have 3' overhangs. Some siRNA agents will have single-stranded overhangs, in one embodiment being 3' overhangs of 1, 2, or 3 nucleotides in length at each end. The overhangs may be the result of one strand being longer than the other, or the result of two strands of equal length being interleaved. The 5' end may be phosphorylated.

[0368] In one embodiment, the length of the double-stranded region is between 15 and 30 nucleotides, or 18, 19, 20, 21, 22, and 23 nucleotides, for example, within the range of siRNA agents discussed above. The siRNA agent is similar in length and structure to the product of natural Dicer processed from long dsiRNA. Embodiments are also included in which the two strands of the siRNA agent are joined, for example, covalently. Hairpin structures or other single-stranded structures providing the desired double-stranded region, as well as 3' overhangs, are also present in this invention.

[0369] The isolated iRNA agents described herein, including dsiRNA agents and siRNA agents, can mediate the silencing of target RNAs, such as mRNAs, or transcripts of genes encoding proteins. For convenience, this mRNA is also referred to herein as the mRNA to be silenced. The gene is also referred to as the target gene. Typically, the RNA to be silenced is an endogenous gene or a pathogen gene. Additionally, RNAs other than mRNAs, such as tRNAs and viral RNAs, can also be targeted.

[0370] As used in this article, the phrase “mediated RNAi” refers to the ability to silence target RNA in a sequence-specific manner. While not wishing to be bound by theory, it is believed that the mechanisms or steps of silencing RNAi and the guide RNA, such as 21 to 23 nucleotide siRNA agents, are involved.

[0371] As used herein, "specifically hybridizable" and "complementary" are terms used to indicate sufficient complementarity to produce stable and specific binding between the compounds of the present invention and the target RNA molecules. Specific binding requires sufficient complementarity to avoid nonspecific binding of the oligomeric compound to non-target sequences under the conditions for desired specific binding, namely, physiological conditions in in vivo assays or therapeutic treatments, or conditions under which the assay is performed in vitro. Non-target sequences typically differ by at least 5 nucleotides.

[0372] In one embodiment, the iRNA agent is "fully complementary" to the target RNA, such as the target mRNA, so that the iRNA agent silences the production of the protein encoded by the target mRNA. In another embodiment, the iRNA agent is "exactly complementary" to the target RNA, for example, the target RNA and the iRNA agent anneal to form, for example, a hybrid uniquely composed of Watson-Crick base pairs in precisely complementary regions. The "fully complementary" target RNA may include an internal region (e.g., having at least 10 nucleotides) that is precisely complementary to the target RNA. Furthermore, in one embodiment, the iRNA agent specifically identifies single nucleotide differences. In this case, if precise complementarity is found in a single nucleotide difference in that region (e.g., within 7 nucleotides), then the iRNA agent mediates RNAi only.

[0373] As used herein, the term “oligonucleotide” refers to a nucleic acid molecule (RNA or DNA) that is, for example, less than 100, 200, 300, or 400 nucleotides in length.

[0374] MicroRNAs (miRNAs) are a highly conserved class of small RNA molecules that are transcribed from DNA in the genomes of plants and animals but are not translated into proteins. Processed miRNAs are single-stranded RNA molecules of approximately 17-25 nucleotides (nt) that are incorporated into the RNA-induced silencing complex (RISC) and have been identified as key regulators of development, cell proliferation, apoptosis, and differentiation. They are believed to play a role in the regulation of gene expression by binding to the 3'-untranslated region of specific mRNAs. RISC mediates the downregulation of gene expression through translational repression, transcript cleavage, or both. RISC is also involved in transcriptional silencing in the nucleus of a wide range of eukaryotic cells.

[0375] In one embodiment, the nucleic acid is an antisense oligonucleotide targeting a target polynucleotide. The term "antisense oligonucleotide" or simply "antisense" is intended to include oligonucleotides complementary to the target polynucleotide sequence. Antisense oligonucleotides are single-stranded DNA or RNA complementary to a selected sequence. In the case of antisense RNA, they prevent translation of the complementary RNA strand by binding to it. Antisense DNA can be used to target a specific complementary (coding or non-coding) RNA. If binding occurs, the DNA / RNA hybrid can be degraded by the enzyme RNase H. In a particular embodiment, the antisense oligonucleotide contains about 10 to about 50 nucleotides, more preferably about 15 to about 30 nucleotides. The term also covers antisense oligonucleotides that may not be precisely complementary to the desired target gene. Therefore, the present invention can be used when non-target-specific activity is found for the antisense, or when containing one or more antisense sequences that mismatch with the target sequence is most preferred for a particular purpose.

[0376] Methods for generating antisense oligonucleotides are known in the art and can be readily modified to produce antisense oligonucleotides targeting any polynucleotide sequence. The selection of antisense nucleotide sequences specific to a given target sequence is based on the analysis of the selected target sequence and the determination of secondary structure, Tm, binding energy, and relative stability. Antisense oligonucleotides can be selected based on the fact that they are relatively incapable of forming dimers, hairpin structures, or other secondary structures that would reduce or prevent specific binding to the target mRNA in the host cell. Highly preferred target regions of mRNA include those regions at or near the AUG translation start codon and those sequences that are substantially complementary to the 5' region of the mRNA. These secondary structure analyses and target site selection considerations can be performed using, for example, OLIGO primer analysis software (Molecular Biology Insights) v.4 and / or BLASTN 2.0.5 algorithm software (Altschul et al., Nucleic Acids Res. 1997, 25(17): 3389-402).

[0377] The oligonucleotides used in this application include both naturally occurring and chemically modified oligonucleotides. Chemical modifications here include nucleoside modifications (including sugar moiety modifications and nucleobase modifications) and inter-nucleoside linkage modifications. Chemical modifications of oligonucleotides do not include those differing only in the nucleobase sequence. "Naturally occurring" here refers to naturally occurring RNA or DNA.

[0378] 3. How to use

[0379] In some aspects, this disclosure provides a method for regulating (e.g., reducing or eliminating) the expression of a target gene in a subject, including administering the aforementioned conjugate to the subject.

[0380] In some aspects, this disclosure provides a method for regulating (e.g., reducing or eliminating) the expression of a target gene in a subject's cells or tissues, including administering the aforementioned conjugate to the subject.

[0381] In some aspects, this disclosure provides a method for delivering a nucleic acid drug to a subject, including administering the conjugate of this disclosure to the subject.

[0382] In some aspects, this disclosure provides a method for treating or preventing a disease in a subject in need, comprising administering to the subject a therapeutically effective amount of the aforementioned conjugate.

[0383] In some respects, this disclosure provides the aforementioned conjugates for regulating (e.g., reducing or eliminating) the expression of target genes in subjects.

[0384] In some respects, this disclosure provides the aforementioned conjugates for regulating (e.g., reducing or eliminating) the expression of target genes in the cells or tissues of a subject.

[0385] In some respects, this disclosure provides the aforementioned conjugate for delivering nucleic acid drugs to a subject.

[0386] In some respects, this disclosure provides the aforementioned conjugates for the treatment or prevention of diseases in subjects who require them.

[0387] In some respects, this disclosure provides for the use of the aforementioned conjugates in the manufacture of medicaments for regulating (e.g., reducing or eliminating) the expression of target genes in subjects.

[0388] In some respects, this disclosure provides the use of the aforementioned conjugates in the manufacture of medicaments that regulate (e.g., reduce or eliminate) the expression of target genes in subject cells or tissues.

[0389] In some respects, this disclosure provides the use of the aforementioned conjugates in the manufacture of pharmaceutical preparations for delivering nucleic acid drugs to subjects.

[0390] In some aspects, this disclosure provides the use of the aforementioned conjugates in the manufacture of a medicament for treating or preventing a disease in a subject of need. In some embodiments, the subject is a cell. In some embodiments, the test substance is a tissue. In some embodiments, the subject is a human being.

[0391] In some implementations, the target genes are factor VII, Eg5, PCSK9, TPX2, apoB, SAA, TTR, HBV, HCV, RSV, PDGF beta gene, Erb-B gene, Src gene, CRK gene, GRB2 gene, RAS gene, MEKK gene, JNK gene, RAF gene, Erk1 / 2 gene, PCNA(p21) gene, MYB gene, JUN gene, FOS gene, BCL-2 gene, Cyclin D gene, VEGF gene, epidermal growth factor receptor gene, Cyclin A gene, Cyclin E gene, WNT-1 gene, beta-catenin gene, c-MET gene, PKC gene, NFKB gene, STAT3 gene, survivin gene, Her2 / Neu gene, topoisomerase I gene, and topoisomerase II. alpha gene, p73 gene, p21(WAF1 / CIP1) gene, p27(KIP1) gene, p27(KIP1) gene, p27(KIP1) gene, PPM1D gene, RAS gene, MIB I gene, MTAI gene, M68 gene, tumor suppressor gene mutation, p53 tumor suppressor gene, LDHA, or any combination thereof.

[0392] In some implementations, administration leads to a reduction or elimination of the expression of the target gene in the subject.

[0393] In some implementations, the administration of the drug leads to a reduction or disappearance of target gene expression in the subject.

[0394] In some implementations, the disease is a viral infection, such as HCV, HBV, HPV, HSV, or HIV infection.

[0395] In some implementations, the disease is cancer.

[0396] In some implementations, the cancers are bile duct cancer, bladder cancer, transitional cell carcinoma, urothelial carcinoma, brain cancer, glioma, astrocytoma, breast cancer, transitional cell carcinoma, cervical cancer, cervical squamous cell carcinoma, rectal cancer, colon cancer, hereditary nonpolyposis colorectal cancer, colorectal adenocarcinoma, gastrointestinal stromal tumor (GIST), endometrial cancer, endometrial stromal sarcoma, esophageal cancer, esophageal squamous cell carcinoma, esophageal adenocarcinoma, ocular melanoma, uveal melanoma, gallbladder cancer, colon cancer, colon cancer, hereditary nonpolyposis colorectal cancer, colorectal adenocarcinoma, gastrointestinal stromal tumor (GIST), endometrial cancer, endometrial stromal sarcoma, gallbladder cancer, gallbladder adenocarcinoma, renal cell carcinoma, clear cell renal cell carcinoma, transitional cell carcinoma, urothelial carcinoma, Wilms' tumor, leukemia, acute lymphoblastic leukemia. Acute myeloid leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic granulocytic leukemia (CMML), liver cancer, liver cancer, hepatocellular carcinoma, cholangiocarcinoma, liver cancer, hepatocellular carcinoma, liver cancer, cholangiocarcinoma, hepatoblastoma, lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, B-cell lymphoma, non-Hodgkin lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, T-cell lymphoma, non-Hodgkin lymphoma, precursor T-cell lymphoma / leukemia, peripheral T-cell lymphoma, multiple myeloma, nasopharyngeal carcinoma (NPC), neuroblastoma, oropharyngeal carcinoma, oral squamous cell carcinoma, osteosarcoma, ovarian cancer, pancreatic cancer, pancreatic ductal adenocarcinoma, pseudopapillary tumor, sharp cell carcinoma. Prostate cancer, prostate adenocarcinoma, skin cancer, melanoma, malignant melanoma, skin melanoma, small intestine cancer, stomach cancer, gastrointestinal stromal tumor (GIST), uterine cancer or uterine sarcoma.

[0397] In some implementation schemes, the cancer is liver cancer, hepatocellular carcinoma, cholangiocarcinoma, or hepatoblastoma.

[0398] In some implementations, the disease is a proliferative, inflammatory, autoimmune, neurological, ocular, respiratory, metabolic, cutaneous, auditory, hepatic, renal, or infectious disease. In some implementations, the disease is a liver disease.

[0399] Example

[0400] Example 1: Preparation of the compound

[0401] 1.1 Preparation of BSQ1

[0402] first step:

[0403] In a 100 mL three-necked flask, add 30 mL of dichloromethane and compound 1 (5.0 g, 23 mmol), then add trifluoroacetic acid (10.5 g, 92 mmol) under ice bath conditions. Finally, react at room temperature for 2 hours. After the reaction is complete, concentrate the solvent under reduced pressure to obtain compound 2, which can be used directly in the next step.

[0404] Step Two:

[0405] 80 mL of DMF was added to a 250 mL three-necked flask, and stirring was started. Under argon protection, compound 2 (3.53 g, 23 mmol), HATU (10.5 g, 27.6 mmol), HOAT (3.4 g, 25 mmol), and compound 3 (8.12 g, 23 mmol) were added in an ice bath. Then, DIPEA (8.9 g, 69 mmol) was added dropwise. After reacting at room temperature for 2 hours, water was added, and the mixture was extracted with ethyl acetate, dried over sodium sulfate, and concentrated to obtain crude compound 4. The crude compound 4 was separated by column chromatography with silica gel (solvent ratio: dichloromethane / methanol = 100 / 1 to 20 / 1) to give 9.6 g of pure compound 4, an off-white solid, with a yield of 92% and a purity of 99.4%.

[0406] 1 H NMR (400MHz, CDCl3) δ7.77-7.79(d, 2H), 7.61-7.63(d, 2H), 7.40-7.44(m, 2 H), 7.30-7.35(m, 2H), 5.25(brs, 1H), 4.26-4.39(m, 2H), 4.21-4.34(m, 2H), 3.74(brs, 1H), 3.67-3.74(m, 5H), 3.61-3.62(m, 1H), 3.19-3.39(m, 4H), 2.2 3-2.33(m, 3H), 1.62-1.67(m, 2H), 1.50-1.57(m, 2H), 1.29-1.46(m, 2H)ppm.

[0407] Step 3:

[0408] Under argon protection, pure compound 4 (3.0 g, 6.6 mmol) was added to a 250 mL single-necked flask, dissolved in 100 mL of pyridine, and then 4,4'-dimethoxytriphenylchloromethane (9.0 g, 26.6 mmol) was added. The reaction was allowed to proceed at room temperature for 1 hour. TLC showed that the reaction was almost complete. The reaction was quenched by adding 10 mL of methanol, and the pyridine was removed by rotary evaporation. The mixture was extracted with 50 mL of ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and passed through a chromatography column (eluent ratio: dichloromethane / methanol = 10 / 1) to give a total of 4.7 g, 6.2 mmol, white solid compound 5.

[0409] Step 4:

[0410] Under argon protection, compound 5 (4.7 g, 6.2 mmol), 50 mL of dichloromethane, 1.3 g, 10 mmol of ethylthiotetraargonazole, and bis(diisopropylamino)(2-cyanoethoxy)phosphine (compound 6) (2.3 g, 7.6 mmol) were added to a 100 mL single-necked flask. The mixture was reacted at room temperature for 2 hours, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and passed through a chromatography column (eluent: ethyl acetate) to give a white solid compound 7 (BSQ1) (3.7 g, 3.9 mmol).

[0411] 1 H NMR (300MHz, DMSO) δ7.89 (d, J=7.5Hz, 2H), 7.69 (d, J=7.3Hz, 2H), 7.44-7.16 (m, 14H), 6.88 (dd, J=8.9, 2 .3Hz, 4H), 4.35-4.16(m, 4H), 3.73(s, 6H), 3.68-3.45(m, 6H), 3.31-3.17(m, 2H), 3.12-2.90(m, 4H), 2.82 -2.62 (m, 2H), 2.16 (d, J=6.9Hz, 2H), 1.54-1.34 (m, 4H), 1.30-1.18 (m, 2H), 1.18-0.97 (m, 12H).

[0412] 31 P NMR (162MHz, DMSO) δ147.43(s), 147.13(s), 146.92(s), 146.62(s).

[0413] 1.2 Preparation of BSQ2

[0414] first step:

[0415] In a 100 mL three-necked flask, add 30 mL of dichloromethane and compound 1 (5 g, 23 mmol), then add trifluoroacetic acid (10.5 g, 92 mmol) under ice bath conditions. Finally, react at room temperature for 2 hours. After the reaction is complete, concentrate the solvent under reduced pressure to obtain compound 2, which can be used directly in the next step.

[0416] Step Two:

[0417] 80 mL of DMF was added to a 250 mL three-necked flask. Stirring was started, and under argon protection, compound 2 (3.53 g, 23 mmol), HATU (10.5 g, 27.6 mmol), HOAT (3.4 g, 25 mmol), and compound 8 (8.85 g, 23 mmol) were added in an ice bath. Then, DIPEA (8.9 g, 69 mmol) was added dropwise, and the reaction was carried out at room temperature for 2 hours. After post-treatment, water was added, and the mixture was extracted with ethyl acetate, dried over sodium sulfate, and concentrated to obtain crude compound 9. The crude compound 9 was separated by column chromatography with silica gel (solvent ratio: dichloromethane / methanol = 100 / 1 to 20 / 1) to give 10.35 g of pure compound 9, an off-white solid, with a yield of 93% and a purity of 99.0%.

[0418] 1 H NMR (400MHz, CDCl3) δ7.77-7.79 (d, 2H), 7.63-7.64 (d, 2H), 7.40-7.44 (m, 2H), 7.30-7.35 (m, 2H), 5 .92-5.96 (m, 1H), 4.08-4.42 (m, 6H), 3.55-3.74 (m, 10H), 3.39-3.50 (m, 5H), 2.20-2.40 (m, 2H)ppm.

[0419] Step 3:

[0420] Under argon protection, compound 9 (2.0 g, 4.1 mmol) was added to a 100 mL single-necked flask, dissolved in 30 mL of pyridine, and 4,4'-dimethoxytriphenylchloromethane (5.6 g, 16.5 mmol) was added. The reaction was allowed to proceed at room temperature for 1 hour, and the reaction was quenched by adding 10 mL of methanol. Pyridine was removed by rotary evaporation, extracted with 50 mL of ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and passed through a chromatography column (eluent ratio: dichloromethane / methanol = 10 / 1) to give a white solid compound 10 (2.5 g, 3.2 mmol).

[0421] Step 4:

[0422] Under argon protection, compound 10 (2.5 g, 3.2 mmol), 30 mL of dichloromethane, 0.7 g, 5.4 mmol of ethylthiotetraargonazole, and 1.2 g, 4 mmol of bis(diisopropylamino)(2-cyanoethoxy)phosphine (compound 6) were added to a 100 mL single-necked flask. The mixture was reacted at room temperature for 2 hours, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and passed through a chromatography column (eluent: ethyl acetate) to give a white solid compound 11 (BSQ2) (2.3 g, 2.3 mmol).

[0423] 1 H NMR (300MHz, DMSO) δ7.90 (t, J=7.6Hz, 2H), 7.69 (d, J=7.4Hz, 2H), 7.48-7.25 (m, 8H), 7.28-7.12 (m, 4H), 6.88 (dd, J=8.9, 2.2Hz, 4H), 4.33-4.16 (m , 4H), 4.11-3.96(m, 2H), 3.73(s, 6H), 3.70-3.37(m, 12H), 3.30-2.95(m, 6H), 2.81-2.61(m, 2H), 1.18(dt, J=11.3, 5.6Hz, 2H), 1.15-0.93(m, 12H).

[0424] 31 P NMR (162MHz, DMSO) δ147.51(s), 147.23(s), 146.98(s), 146.74(s).

[0425] Example 2: Preparation of Q1-Justice Chain and Q2-Justice Chain

[0426] 2.1 Preparation of Q1-Justice Chain

[0427] first step:

[0428] The method employs a phosphorus amide solid-phase synthesis, starting with a CPG or PS solid-phase support attached to Unylinker (catalog number: C3001-100; manufacturer: Dinaxinke). Following the nucleotide sequence, nucleoside monomers or BSQ1 molecules are sequentially linked from the 3'-5' direction. Each linkage of a nucleoside or BSQ1 involves four steps: deprotection, coupling, capping, oxidation, or thiolation, yielding a Q1-positive chain with a solid-phase support.

[0429] The conditions for each reaction step are as follows:

[0430] (1) Nucleoside monomer or BSQ1: Dissolve in anhydrous acetonitrile to form an acetonitrile solution with a concentration of 0.1 mol / L.

[0431] (2) Deprotection: Add 3% dichloroacetic acid-dichloromethane solution.

[0432] (3) Coupling reaction: Add 0.3 mol / L ETT acetonitrile solution.

[0433] (4) Oxidation reaction: Add 0.05 mol / L of tetrahydrofuran / pyridine / water (70 / 20 / 10, v / v / v) solution of iodine.

[0434] (5) Thio-reaction: Add 0.2 mol / L of hydrogenated xanthan pyridine solution.

[0435] (6) Capping reaction: Add 20% acetic anhydride-acetonitrile and pyridine / N-methylimidazolium / acetonitrile (10 / 14 / 76, v / v / v) solution.

[0436] Step Two:

[0437] The synthesized Q1-positive chain with a solid support was added to a 2 ml centrifuge tube, and 25-28% ammonia solution was added. The mixture was reacted at 55°C for 16 hours, filtered, and then washed three times with 1 mL of 50% ethanol aqueous solution to remove the solid support. The filtrate was concentrated and dried to obtain crude Q1 single-chain product, awaiting purification. The crude Q1 single-chain product was dissolved in 1 ml of RNase-free water and purified by ion-pair reversed-phase chromatography or ion-exchange chromatography. The collected samples were analyzed, and the qualified samples were combined and desalted to obtain pure Q1 single-chain product containing amino modification (compound 12). In this compound, RA is the positive chain, with its 3' end linked to Q1.

[0438] Step 3:

[0439] Compound 12 is a single-chain pure product containing an amino-modified Q1 and requires further coupling to obtain an effective positive-chain sequence containing the GalNAc ligand. Compound 13 was dissolved in anhydrous DMSO and then added to a sodium phosphate buffer solution of compound 12. The mixture was sonicated and vortexed until completely dissolved, and reacted at 25°C for 16 hours. Finally, an equal volume of 25-28% concentrated ammonia was added, and the reaction was carried out at 50°C for 2 hours. After concentration and drying, the product was dissolved in 1 ml of RNase-free water and purified by ion-pair reversed-phase chromatography or ion-exchange chromatography. The collected samples were analyzed, and qualified samples were combined and desalted to obtain compound 14, a pure product containing an effective positive-chain Q1. In this compound, RA is an oligonucleotide (e.g., the positive chain) with its 3' end linked to Q1.

[0440] 2.2 Preparation of Q2-Justice Chain

[0441] first step:

[0442] The method employs a phosphorus amide solid-phase synthesis, starting with a CPG or PS solid-phase support attached to Unylinker (catalog number: C3001-100; manufacturer: Dinaxinke). Following the nucleotide sequence, nucleoside monomers or BSQ2 molecules are sequentially linked from the 3'-5' direction. Each attachment of a nucleoside or BSQ2 molecule involves four steps: deprotection, coupling, capping, oxidation, or thiolation, yielding a Q2-positive chain with a solid-phase support.

[0443] The conditions for each reaction step are as follows:

[0444] (1) Nucleoside monomer or BSQ2: Dissolve in acetonitrile solution with a concentration of 0.1 mol / L.

[0445] (2) Deprotection: Add 3% dichloroacetic acid-dichloromethane solution.

[0446] (3) Coupling reaction: Add 0.3 mol / L ETT acetonitrile solution.

[0447] (4) Oxidation reaction: Add 0.05 mol / L of tetrahydrofuran / pyridine / water (70 / 20 / 10, v / v / v) solution of iodine.

[0448] (5) Thio-reaction: Add 0.2 mol / L of hydrogenated xanthan pyridine solution.

[0449] (6) Capping reaction: Add 20% acetic anhydride-acetonitrile and pyridine / N-methylimidazolium / acetonitrile (10 / 14 / 76, v / v / v) solution.

[0450] Step Two:

[0451] The synthesized Q2-positive chain with a solid support was added to a 2 ml centrifuge tube, and 25-28% ammonia solution was added. The mixture was reacted at 55°C for 16 hours, filtered, and then washed three times with 1 mL of 50% ethanol aqueous solution to remove the solid support. The filtrate was concentrated and dried to obtain crude Q2 single-chain product, awaiting purification. The crude Q2 single-chain product was dissolved in 1 ml of RNase-free water and purified by ion-pair reversed-phase chromatography or ion-exchange chromatography. The collected samples were analyzed, and the qualified samples were combined and desalted to obtain pure Q2 single-chain product containing amino modification (compound 15). In this compound, RA is the positive chain, with its 3' end linked to Q2.

[0452] Step 3:

[0453] Compound 15 is a single-chain pure product of Q2 containing an amino-modified amino group. Further conjugation is required to obtain the effective positive-chain sequence containing the GalNAc ligand. Compound 13 was dissolved in anhydrous DMSO and then added to a sodium phosphate buffer solution of compound 15. The mixture was sonicated and vortexed until completely dissolved, and reacted at 25°C for 16 hours. Finally, an equal volume of 25-28% concentrated ammonia was added, and the reaction was carried out at 50°C for 2 hours. After concentration and drying, the product was dissolved in 1 ml of RNase-free water and purified by ion-pair reversed-phase chromatography or ion-exchange chromatography. The collected samples were analyzed, and qualified samples were combined and desalted to obtain the effective Q2-positive-chain pure product, compound 16. In this product, RA is an oligonucleotide (e.g., the positive chain), with its 3' end linked to Q2.

[0454] Example 3: Preparation of antisense siRNA

[0455] first step:

[0456] The method employed is a phosphorus amide solid-phase synthesis. Starting with a CPG or PS solid-phase support coupled with Unylinker (catalog number: C3001-100; manufacturer: Dinaxinke), the cycle proceeds sequentially, linking nucleoside monomers one by one from the 3'-5' direction according to the nucleotide sequence. Each nucleoside linkage involves four steps: deprotection, coupling, capping, and oxidation or thiolation, yielding an antisense siRNA strand with the solid-phase support. The conditions for each reaction step are as follows:

[0457] (1) Nucleoside monomer: Dissolved in acetonitrile solution with a concentration of 0.1 mol / L.

[0458] (2) Deprotection: Add 3% dichloroacetic acid-dichloromethane solution.

[0459] (3) Coupling reaction: Add 0.3 mol / L ETT acetonitrile solution.

[0460] (4) Oxidation reaction: Add 0.05 mol / L of tetrahydrofuran / pyridine / water (70 / 20 / 10, v / v / v) solution of iodine.

[0461] (5) Thio-reaction: Add 0.2 mol / L of hydrogenated xanthan pyridine solution.

[0462] (6) Capping reaction: Add 20% acetic anhydride-acetonitrile and pyridine / N-methylimidazolium / acetonitrile (10 / 14 / 76, v / v / v) solution.

[0463] Step Two:

[0464] The synthesized siRNA antisense strand with a solid-phase carrier was added to a 2 ml centrifuge tube, and 25-28% ammonia solution was added. The mixture was reacted at 55°C for 16 hours, filtered, and then washed three times with 1 mL of 50% ethanol aqueous solution to remove the solid-phase carrier. The filtrate was concentrated and dried to obtain crude siRNA antisense strand, which was then awaited for purification. The crude siRNA antisense strand was dissolved in 1 ml of RNase-free water and purified by ion-pair reversed-phase chromatography or ion-exchange chromatography. The collected samples were analyzed, and the qualified samples were combined and desalted to obtain pure siRNA antisense strand.

[0465] Example 4: Preparation of siRNA double strands of Q1 and Q2

[0466] The pure Q1-positive strand or the pure Q2-positive strand was mixed with the siRNA antisense strand prepared in Example 3 at a certain molar ratio, denatured at 90 degrees for 3 minutes, then cooled to room temperature, and finally lyophilized to obtain effective pure Q1 double strand and pure Q2 double strand.

[0467] Example 5: Scale-up synthesis of Q1 and Q2

[0468] 5.1 Amplified Synthesis of Q1-Chain of Justice

[0469] first step:

[0470] The steps are the same as the first step in Example 1.1.

[0471] Step Two:

[0472] The steps are the same as step 2 in Example 1.1.

[0473] Step 3:

[0474] In a 100 mL three-necked flask, 30 mL of dichloromethane and compound 4 (5 g, 11 mmol) were added, followed by 15 mL of diethylamine. The reaction was carried out at room temperature for 14 h. After the reaction was completed, the solvent was removed by concentration under reduced pressure. The mixture was then slurried once with methyl tert-butyl ether, the supernatant was discarded, and the mixture was finally dried by pumping to obtain the intermediate (2.65 g), which was used directly in the next step.

[0475] Add 60 mL of LDM to a 250 mL three-necked flask, start stirring, and under nitrogen protection, add compound 17 (4.92 g, 11 mmol), EDCI (2.87 g, 15 mmol), HOBT (1.62 g, 12 mmol), and intermediate (2.65 g, 11 mmol) in an ice bath. Then add DIPEA (2.84 g, 22 mmol) dropwise, and finally react at room temperature for 4 h.

[0476] Post-treatment involved adding water, extraction with ethyl acetate, drying with sodium sulfate, and concentration to obtain the crude product. The crude product was then separated by column chromatography with silica gel (dichloromethane / methanol = 50 / 1 to 15 / 1) to give 5.1 g of compound 18 as an off-white solid, with a yield of 70% and a purity of 98.3%.

[0477] Step 4:

[0478] Add 30 mL of anhydrous pyridine to a 100 mL three-necked flask, start stirring, and under nitrogen protection, add compound 18 (3 g, 4.55 mmol), triethylamine (0.69 g, 6.8 mmol), followed by DMTrCl (1.85 g, 5.46 mmol), and finally react at room temperature for 20 h.

[0479] Post-processing: The reaction was quenched with 1 g of methanol, and the reaction solution was concentrated under reduced pressure. The concentrate was then washed three times with 60 mL of ethyl acetate, followed by three washes with 20 mL of saturated sodium bicarbonate each time, and then once with 20 mL of brine. The solution was dried over sodium sulfate and concentrated to obtain the crude product. The crude product was separated by column chromatography with silica gel (dichloromethane / methanol / triethylamine = 100 / 1 / 0.2 to 50 / 1 / 0.1) to give 3.3 g of compound 19 as a pale yellow solid, with a yield of 75% and a purity of 98.5%.

[0480] Step 5:

[0481] Under argon protection, 2 g of compound 19, 30 mL of dichloromethane, 0.5 g of ethylthiotetraargonazole, and 1 g of bis(diisopropylamino)(2-cyanoethoxy)phosphine (compound 6) were added to a 100 mL single-necked flask. After reacting at room temperature, the mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and passed through a chromatography column (developing solvent: ethyl acetate) to give a white solid compound 20.

[0482] Step 6:

[0483] Repeating the first and second steps of Example 2.1, we obtained compound 16 with a Q2-positive chain structure.

[0484] 5.2 Magnified Synthesis of Effective Q2-Justice Chain

[0485] first step:

[0486] The steps are the same as the first step in Example 1.2.

[0487] Step Two:

[0488] The steps are the same as step 2 in Example 1.2.

[0489] Step 3:

[0490] In a 100 mL three-necked flask, 30 mL of dichloromethane and compound 9 (5 g, 10.3 mmol) were added, followed by 15 mL of diethylamine. The reaction was carried out at room temperature for 14 h. After the reaction was completed, the solvent was removed by concentration under reduced pressure. The mixture was then slurried once with methyl tert-butyl ether, the supernatant was discarded, and the mixture was finally dried by pumping to obtain the intermediate (2.8 g), which was used directly in the next step.

[0491] Add 60 mL of LDM to a 250 mL three-necked flask, start stirring, and under nitrogen protection, add compound 17 (4.92 g, 11 mmol), EDCI (2.87 g, 15 mmol), HOBT (1.62 g, 12 mmol) and intermediate (2.8 g, 10.7 mmol) in an ice bath. Then add DIPEA (2.84 g, 22 mmol) dropwise, and finally react at room temperature for 4 h.

[0492] Post-treatment involved adding water, extraction with ethyl acetate, drying with sodium sulfate, and concentration to obtain the crude product. The crude product was then separated by column chromatography with silica gel (dichloromethane / methanol = 50 / 1 to 15 / 1) to give 5.3 g of compound 21 as an off-white solid, with a yield of 68% and a purity of 97.8%.

[0493] Step 4:

[0494] Add 30 mL of anhydrous pyridine to a 100 mL three-necked flask, start stirring, and under nitrogen protection, add compound 21 (3.15 g, 4.55 mmol), triethylamine (0.69 g, 6.8 mmol), followed by DMTrCl (1.85 g, 5.46 mmol), and finally react at room temperature for 20 h.

[0495] Post-processing: The reaction was quenched with 1 g of methanol, and the reaction solution was concentrated under reduced pressure. The concentrate was then washed three times with 60 mL of ethyl acetate, followed by washing three times with 20 mL of saturated sodium bicarbonate each time, and then once with 20 mL of brine. The solution was dried over sodium sulfate and concentrated to obtain the crude product. The crude product was separated by column chromatography with silica gel (dichloromethane / methanol / triethylamine = 100 / 1 / 0.2 to 50 / 1 / 0.1) to give 3.4 g of compound 22 as a pale yellow solid, with a yield of 75.5% and a purity of 97.3%.

[0496] Step 5:

[0497] Under argon protection, 2 g of compound 22, 30 ml of dichloromethane, 0.5 g of ethylthiotetraargonazole, and 1 g of bis(diisopropylamino)(2-cyanoethoxy)phosphine (compound 6) were added to a 100 mL single-necked flask. After reacting at room temperature, the mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and passed through a chromatography column (developing solvent: ethyl acetate) to obtain a white solid, compound 23.

[0498] Step 6:

[0499] Repeating the first and second steps of Example 2.2, we obtained compound 16 with a Q2-positive chain structure.

[0500] Example 6: Primary Mouse Hepatocyte Experiment

[0501] 6.1: Compound Information

[0502] The Q1 and Q2 bichains TTR-Q1 and TTR-Q2 containing the following sense and antisense chains were prepared according to the methods of Examples 2, 3 and 4.

[0503] The positive chains of TTR-Q1 and TTR-Q2 are shown in SEQ ID NO:1. Specifically, the positive chain sequence is: AmsAmsCmAmGmUmGfUmUfCfUfUmGmCmUmCmUmAmUmAm (SEQ ID No: 1). Wherein, Am = 2'-O-methoxyadenosine-3'-phosphate, Ams = 2'-O-methoxyadenosine-3'-thiophosphate, Af = 2'-fluoroadenosine-3'-phosphate, Afs = 2'-fluoroadenosine-3'-thiophosphate, Gm = 2'-O-methoxyguanosine-3'-phosphate, Gms = 2'-O-methoxyguanosine-3'-thiophosphate, Gf = 2'-fluoroguanosine-3'-phosphate, Gfs = 2'-fluoroguanosine-3'-thiophosphate. Ester, Cm = 2'-O-methoxycytidine-3'-phosphate, Cms = 2'-O-methoxycytidine-3'-thiophosphate, Cf = 2'-fluorocytidine-3'-phosphate, Cfs = 2'-fluorocytidine-3'-thiophosphate, Um = 2'-O-methoxyuridine-3'-phosphate, Ums = 2'-O-methoxyuridine-3'-thiophosphate, Uf = 2'-fluorouridine-3'-phosphate, Ufs = 2'-fluorouridine-3'-thiophosphate.

[0504] The antisense strand sequences of TTR-Q1 and TTR-Q2 are shown in SEQ ID No: 2. Specifically, the antisense strand sequence is: UmsUfsAmUmAmGfAmGfCfAmAmGmAmAfCmAfCmAfCmUmGmUmUmsUmsUm (SEQ ID No: 2). Wherein, Am = 2'-O-methoxyadenosine-3'-phosphate, Ams = 2'-O-methoxyadenosine-3'-thiophosphate, Af = 2'-fluoroadenosine-3'-phosphate, Afs = 2'-fluoroadenosine-3'-thiophosphate, Gm = 2'-O-methoxyguanosine-3'-phosphate, Gms = 2'-O-methoxyguanosine-3'-thiophosphate, Gf = 2'-fluoroguanosine-3'-phosphate, Gfs = 2'-fluoroguanosine-3'-thiophosphate. Ester, Cm = 2'-O-methoxycytidine-3'-phosphate, Cms = 2'-O-methoxycytidine-3'-thiophosphate, Cf = 2'-fluorocytidine-3'-phosphate, Cfs = 2'-fluorocytidine-3'-thiophosphate, Um = 2'-O-methoxyuridine-3'-phosphate, Ums = 2'-O-methoxyuridine-3'-thiophosphate, Uf = 2'-fluorouridine-3'-phosphate, Ufs = 2'-fluorouridine-3'-thiophosphate.

[0505] TTR-Q1 is a double-stranded compound containing compound 14 (where RA is SEQ ID No: 1, and its 3' end is attached to Q1) and an antisense strand (SEQ ID No: 2). TTR-Q2 is a double-stranded compound containing compound 16 (where RA is SEQ ID No: 1, and its 3' end is attached to Q2) and an antisense strand (SEQ ID No: 2). TTR-L96 is a double-stranded compound of SEQ ID No: 1 and SEQ ID No: 2, with the 3' end of SEQ ID No: 1 attached to L96, wherein the structure of L96 is as follows:

[0506] 6.2: Experimental Objective:

[0507] Primary hepatocytes were isolated from ordinary C57BL / 6 mice and transfected with the compound that allows free uptake. The inhibitory activity of the compound on the TTR gene was detected by qPCR, and the activity of TTR protein in the supernatant was detected by ELISA.

[0508] 6.3 Primers, as shown in Table 1 below.

[0509] Table 1 Primers for Examples

[0510] 6.4 Reagents and materials are shown in Table 2 below.

[0511] Table 2 Reagents and Materials

[0512] 6.5: Compound Preparation

[0513] Compounds TTR-Q1, TTR-Q2, and TTR-L96 were dissolved in DEPC-containing water at a ratio of 50 μL DEPC H2O per OD, and their concentrations were determined using a Nanodrop 2000. Based on the measurement results, the compounds were diluted to 266 ng / μL and used as stock solutions, which were then stored at -20°C. Specific dilution methods for in vitro activity assays of the compounds are shown in Table 3.

[0514] Table 3: Dilution methods for freely available compounds

[0515] 6.6: Sample Preparation

[0516] On the first day of the experiment, primary mouse hepatocytes were isolated, and the compound was delivered using a free uptake method without transfection reagents. Simply add 1 μL of the appropriate concentration of compound per well directly to the cell seeding plate.

[0517] 8000 cells per well were seeded into 96-well plates. Starting with the highest concentration of 10 nM, eight concentrations were obtained through 4-fold dilutions for free uptake. After drug administration, the plates were incubated at 37°C with 5% CO2.

[0518] 6.7: RNA Extraction and Reverse Transcription

[0519] Total RNA extraction from cells: Cells were lysed and total RNA was extracted using a high-throughput nucleic acid extractor-magnetic bead method.

[0520] The instrument operating parameters are as follows:

[0521] Table 4: Operating Procedures for High-Throughput Nucleic Acid Extractors

[0522] RNA concentration adjustment: Sample concentration was measured using a nanophotometer, and water was added to adjust all samples to the same concentration. Reverse transcription: All samples were reverse transcribed into cDNA using a reverse transcription kit.

[0523] 6.8 Real-time quantitative PCR detection

[0524] Standard real-time quantitative qPCR detection: cDNA samples are relatively quantified using a standard qPCR kit. Each cDNA sample is analyzed in triplicate.

[0525] 6.9 Data Processing and Analysis

[0526] Calculate the difference factor: Fc = 2^ -△△Cp .

[0527] Plotting: Analyzing IC using "[Inhibitor] vs. normalized response -- Variable slope" in Graphpad Prism 7 50 .

[0528] (Y=100 / (1+(X^HillSlope) / (IC 50 ^HillSlope)))

[0529] The results were summarized using GraphPad Prism 7 and are shown in Figure 3. Based on the inhibition efficiency results, the free-uptake group showed that both TTR-Q1 and TTR-Q2 at various concentrations could successfully deliver TTR-Q1 to primary mouse hepatocytes and achieve gene silencing. Furthermore, their effect was superior to TTR-L96.

[0530] Example 7: Study on the long-term effect of drugs in mice

[0531] The siRNA conjugates detected were ApoC3-Q1, ApoC3-Q2, and ApoC3-L96.

[0532] The positive chains of ApoC3-Q1 and ApoC3-Q2 are shown in SEQ ID NO:3. Specifically, the positive chain sequence is: CfsCmsUfCmCfCmAfAmUfAfAfAmGfCmUfGmGfAmCfAmAf (SEQ ID No: 3). Wherein, Am = 2'-O-methoxyadenosine-3'-phosphate, Ams = 2'-O-methoxyadenosine-3'-thiophosphate, Af = 2'-fluoroadenosine-3'-phosphate, Afs = 2'-fluoroadenosine-3'-thiophosphate, Gm = 2'-O-methoxyguanosine-3'-phosphate, Gms = 2'-O-methoxyguanosine-3'-thiophosphate, Gf = 2'-fluoroguanosine-3'-phosphate, Gfs = 2'-fluoroguanosine-3'-thiophosphate. Ester, Cm = 2'-O-methoxycytidine-3'-phosphate, Cms = 2'-O-methoxycytidine-3'-thiophosphate, Cf = 2'-fluorocytidine-3'-phosphate, Cfs = 2'-fluorocytidine-3'-thiophosphate, Um = 2'-O-methoxyuridine-3'-phosphate, Ums = 2'-O-methoxyuridine-3'-thiophosphate, Uf = 2'-fluorouridine-3'-phosphate, Ufs = 2'-fluorouridine-3'-thiophosphate.

[0533] The antisense strand sequences of ApoC3-Q1 and ApoC3-Q2 are shown in SEQ ID No: 4. Specifically, the antisense strand sequence is: UmsUfsGmUfCmCfAmGfCmUfUmUmAmUfUmGfGmGfAmGfGmsCmsCm (SEQ ID No: 4). Wherein, Am = 2'-O-methoxyadenosine-3'-phosphate, Ams = 2'-O-methoxyadenosine-3'-thiophosphate, Af = 2'-fluoroadenosine-3'-phosphate, Afs = 2'-fluoroadenosine-3'-thiophosphate, Gm = 2'-O-methoxyguanosine-3'-phosphate, Gms = 2'-O-methoxyguanosine-3'-thiophosphate, Gf = 2'-fluoroguanosine-3'-phosphate, Gfs = 2'-fluoroguanosine-3'-thiophosphate. Ester, Cm = 2'-O-methoxycytidine-3'-phosphate, Cms = 2'-O-methoxycytidine-3'-thiophosphate, Cf = 2'-fluorocytidine-3'-phosphate, Cfs = 2'-fluorocytidine-3'-thiophosphate, Um = 2'-O-methoxyuridine-3'-phosphate, Ums = 2'-O-methoxyuridine-3'-thiophosphate, Uf = 2'-fluorouridine-3'-phosphate, Ufs = 2'-fluorouridine-3'-thiophosphate.

[0534] ApoC3-Q1 is a double-stranded compound containing compound 14 (where RA is SEQ ID No: 3, and its 3' end is attached to Q1) and an antisense strand (SEQ ID No: 4). ApoC3-Q2 is a double-stranded compound containing compound 16 (where RA is SEQ ID No: 3, and its 3' end is attached to Q2) and an antisense strand (SEQ ID No: 4). ApoC3-L96 is a double-stranded compound of SEQ ID No: 3 and SEQ ID No: 4, with the 3' end of SEQ ID No: 3 attached to L96.

[0535] Male ApoC3 humanized mice were grouped according to serum protein expression. Twenty-four mice with relatively consistent expression levels were selected and randomly assigned to other groups. The dosage for each animal was calculated based on body weight, and the medication was administered subcutaneously as a single dose. The siRNA conjugate was administered at a concentration of 3 mg / kg in PBS (using PBS as the solvent). Specifically, before the experiment, the siRNA conjugate was dissolved in PBS and brought to the required concentration and volume. The administration volume for the PBS group (control group) and the siRNA conjugate was 10 mL / kg.

[0536] Blood samples were collected before administration (day 0) and on days 7, 10, 14 and 21 after administration. Serum was separated and ApoC3 protein in the serum was detected using an ELISA kit (abcam, ab154131).

[0537] Data statistics and analysis

[0538] Calculate the ApoC3 protein concentration in the drug-treated group and the control group and convert it into a percentage to obtain the residual protein activity;

[0539] Normalize the protein concentrations of each group on Day 0 and use Graphpad Prism 7 to generate time-lapse graphs.

[0540] The results are shown in Figure 4, and the specific dosing regimens are shown in Table 5 below.

[0541] Table 5. Compound information and dosing regimens for different groups.

[0542] The results showed that both ApoC3-Q1 and ApoC3-Q2 could effectively silence the ApoC3 protein, and their effects were superior to those of ApoC3-L96. Furthermore, the long-term effects of ApoC3-Q1 and ApoC3-Q2 were better than those of ApoC3-L96.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, having the general structural formula shown in formula (IL): in, Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; X1 is a branch; X2 is a nucleophilic or electrophilic group; R1-R6 are any substituents.

2. The compound of claim 1, further comprising W, having the molecular formula shown in (IW). W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino-substituted group or amino-protecting group optionally substituted with one or more halogens.

3. The compound of claim 1, further comprising ligand X, having the molecular formula shown in (IX): in, X is a ligand or a derivative thereof.

4. The compound according to any one of claims 1-3, wherein, R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 Cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

5. The compound according to any one of claims 1-4, wherein, X1 includes -(CH2) n -、-(CH2) n NH-, -(CH2) n O-, -(CH2) n S-、-R X12 (CH2) n -、-R X12 (CH2) n NH-, -R X12 (CH2) n O- or -R X12 (CH2) n One of the S-; Where R X12 It is one of N, O, and S; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

6. The compound according to any one of claims 1-5, wherein, X1 includes -(CH2) n -、-(CH2) n NH-, -(CH2) n O-, -(CH2) n S-、-R X12 (CH2) n -、-R X12 (CH2) n N-、-R X12 (CH2) n O- or -R X12 (CH2) n S- optional substitution; wherein the substituents optionally substituted are selected from the group consisting of: halogen, cyano, hydroxy, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C3 alkyl polyoxyethylene, or C1-C3 alkyl polyoxypropylene; Where R X12 It is one of N, O, and S; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

7. The compound according to any one of claims 1-6, wherein, X1 includes -((CH2) m1 O) m2 -,-((CH2) m1 O) m2 CH2-, -CH2((CH2) m1 O) m2 CH2-, m1 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5.

8. The compound according to any one of claims 1-7, wherein, X1 includes -((CH2) m1 O) m2 -,-((CH2) m1 O) m2 CH2-, -CH2((CH2) m1 O) m2 Optional substitution of CH2- The substituents optionally substituted may be selected from the group consisting of: halogen, cyano, hydroxy, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C3 alkyl polyoxyethylene, or C1-C3 alkyl polyoxypropylene; Where R X12 It is one of N, O, and S; m1 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5.

9. The compound according to any one of claims 1-8, wherein, Compounds of formula (IL), formula (IW), or formula (IX), or pharmaceutically acceptable salts thereof, further include one of the following structures: Where R1-R 12 Including hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-6 alkyl, optional terminating C 1-6 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-6 alkyl carbonyl, C with optional substituent 1-6 One of cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene. n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-; X is a ligand or a derivative thereof; W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino-substituted group or amino-protecting group optionally substituted with one or more halogens.

10. The compound according to any one of claims 1-9, wherein X2 comprises one of -NH2, -SH, -OH, -NH-, -O-, or -S-.

11. The compound as described in any one of claims 1-10, wherein, Compounds of formula (IL), (IW), or (IX), or pharmaceutically acceptable salts thereof, further have one of the following structures: Where R1-R 12 Including hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-6 alkyl, optional terminating C 1-6 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-6 alkyl carbonyl, C with optional substituent 1-6 One of cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene. W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino substituent or amino protecting group optionally substituted by one or more halogens; X is a ligand or a derivative thereof; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino-substituted group or amino-protecting group optionally substituted with one or more halogens.

12. The compound according to any one of claims 1-11, wherein the compound of formula (I1), formula (IW) or formula (IX) or a pharmaceutically acceptable salt thereof further has one of the following structures: Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; X1 is a branch; X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-; W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino substituent or amino protecting group optionally substituted by one or more halogens; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; in, R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 Cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

13. The compound according to any one of claims 1-12, wherein the compound of formula (I1), formula (IW) or formula (IX) or a pharmaceutically acceptable salt thereof further has one of the following structures: Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; X1 is a branch; X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-; W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino substituent or amino protecting group optionally substituted by one or more halogens; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; in, R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 Cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

14. The compound according to any one of claims 1-13, wherein the compound of formula (I1), formula (IW) or formula (IX) or a pharmaceutically acceptable salt thereof further has one of the following structures: Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; X1 is a branch; X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-; W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino substituent or amino protecting group optionally substituted by one or more halogens; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; in, R5-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 Cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

15. The compound as described in any one of claims 1-14, wherein X is selected from: galactose, galactosamine, N-acetylgalactosamine, mannose, glucose, glucosamine, N-acetylglucosamine, fucose or lactose, N-acetylgalactosamine with hydroxyl groups fully protected by an acyl group, galactose with hydroxyl groups fully protected by an acyl group, galactosamine with hydroxyl groups fully protected by an acyl group, N-formyl-galactosamine with hydroxyl groups fully protected by an acyl group, N-propionyl-galactosamine with hydroxyl groups fully protected by an acyl group, N-butyryl-galactosamine with hydroxyl groups fully protected by an acyl group, or N-isobutyryl-galactosamine with hydroxyl groups fully protected by an acyl group, wherein the acyl group is an acetyl group or a benzoyl group.

16. The compound of claim 15, wherein X is a lipid or a lipid-based molecule.

17. The compound of claim 15, wherein the molecular formula of X is selected from one of the following: Where r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

18. The compound as shown in any one of claims 2-17, wherein the compound of formula (IX) or a pharmaceutically acceptable salt thereof further has one of the following structures. Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; X1 is a branch; X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; in, R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 Cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

19. The compound according to any one of claims 1-18, wherein the compound of formula (IW) or formula (IX) or a pharmaceutically acceptable salt thereof further has the following general formula. Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; X is a ligand or a derivative thereof; W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino-substituted group or amino-protecting group optionally substituted with one or more halogens.

20. The compound according to any one of claims 1-19, wherein the compound of formula (IW) or a pharmaceutically acceptable salt thereof further has one of the following structures: Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; in, R5-R 12 Selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 Cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

21. The compounds of claims 1-20, wherein the compound of formula (IX) or a pharmaceutically acceptable salt thereof further has one of the following structures. Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; r is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5.

22. A scaffold structure comprising the compound as described in claims 1-21, comprising one of the following structures: Where q is an integer from 1 to 3, Y includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; Z includes one of hydrogen, a hydroxyl protecting group, a phosphorus-containing reactive group, a C1-C6 alkyl group optionally substituted with one or more halogens, a phosphate group, or a substituted acyl group; X1 is a branch; X2 is a nucleophilic or electrophilic group; X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-; W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino substituent or amino protecting group optionally substituted by one or more halogens; in, R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 Cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

23. The support structure as described in claim 22, further comprising one of the following structures: X1 is a branch; X2 is a nucleophilic or electrophilic group; X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-; W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl optionally substituted with one or more halogens, amino substituents, or amino protecting groups. in, R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 Cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; Where q is an integer from 1 to 3, The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

24. The support structure as shown in any one of claims 22-23, having the following general formula: X1 is a branch; X2 is a nucleophilic or electrophilic group; X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-; W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino substituent or amino protecting group optionally substituted by one or more halogens; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; in, R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 Cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

25. The scaffold structure as described in any one of claims 22-24, further comprising one having the following chemical formula: n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5.

26. The support structure as described in any one of claims 22-25, further comprising one of the following structures: n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5.

27. A conjugate or a pharmaceutically acceptable salt thereof, wherein the conjugate comprises: (i) One or more oligonucleotides; (ii) One or more ligands X; and (iii) one or more linker units L, wherein the conjugate structure includes one of the following structures: Where X is a ligand or its derivative, and RA is an oligonucleotide.

28. A conjugate having the following general structural formula: in, L is the connector unit L as described in any one of claims 1-21; X is a ligand or a derivative thereof; W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino substituent or amino protecting group optionally substituted by one or more halogens; RA stands for oligonucleotide; p is 0-10, preferably 1-3, and more preferably 2.

29. The compound conjugate of claim 28, further comprising one of the following structures: X1 is a branch; X2 is a nucleophilic or electrophilic group; X2 includes one of -NH2, -SH, -OH, -NH-, -O-, or -S-; W includes hydrogen, amino, hydroxyl, thiol, C1-C6 alkyl, amino substituent or amino protecting group optionally substituted by one or more halogens; n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; in, R1-R6 are selected from: hydrogen, halogen, hydroxyl, methoxy, ethoxy, alkylamino, cycloalkylamino, heterocyclic, aryl, heteroaryl, C 1-3 Alkyl polyoxyethylene, C with optional substituents 1-3 alkyl, optional terminating C 1-5 Cycloalkyl, heterocycloalkyl with optional substituent, heterocycloalkylalkyl with optional substituent, aryl with optional substituent, C with optional substituent 1-3 alkyl carbonyl, C with optional substituent 1-5 Cycloalkyl carbonyl, heterocycloalkyl carbonyl with optional substituent, heterocycloalkylalkyl carbonyl with optional substituent, aryl carbonyl with optional substituent, or heteroaryl carbonyl with optional substituent; The groups with optional substituents include halogen, cyano, hydroxyl, nitro, amino, alkylamino, cycloalkylamino, heterocyclic, aminocarbonyl, sulfonyl, aminosulfonyl, carbonylamino, sulfonylamino, methyl, ethyl, aryl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, trifluoromethoxy, trifluoroethoxy, polyoxyethylene, polyoxypropylene, C1-C6 alkyl polyoxyethylene, or one of C1-C6 alkyl polyoxypropylene.

30. The scaffold structure as described in any one of claims 28-29, further comprising one having the following chemical formula: n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; RA stands for oligonucleotide.

31. The support structure as described in any one of claims 28-30, further comprising one of the following structures: n is an integer from 0 to 20, preferably 1 to 10, and more preferably 1 to 5; m2 is an integer from 0 to 20, preferably from 1 to 10, and more preferably from 1 to 5; RA stands for oligonucleotide.

32. The conjugate according to any one of claims 28-31, wherein the oligonucleotide comprises a single-stranded oligonucleotide or a double-stranded oligonucleotide.

33. The conjugate according to any one of claims 28-32, wherein the oligonucleotide comprises one or more modified nucleotides.

34. The conjugate as claimed in claim 33, wherein, The one or more 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′-fluorine modified nucleotides, 2′-deoxy modified nucleotides, and 2′-hydroxy modified nucleotides.

35. The conjugate according to any one of claims 28-34, wherein the oligonucleotide is siRNA.

36. The compound of claim 35, wherein the siRNA comprises a sense strand and an antisense strand that complement each other to form a double helix.

37. A pharmaceutical composition comprising the compound, scaffold, or conjugate of the preceding claims, and optionally a pharmaceutically acceptable carrier.

38. A kit comprising the compound, scaffold, or conjugate of the preceding claims.

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