Integrin ligand and use thereof

Abstract

The present invention relates to an integrin ligand and the use thereof. Specifically, provided is a compound represented by formula I or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or an αvβ6 integrin ligand comprising the compound represented by formula I or the stereoisomer thereof or the pharmaceutically acceptable salt thereof. The compound or the αvβ6 integrin ligand has a good serum stability and affinity for integrin αvβ6, and can be used for delivering a cargo molecule (such as an RNAi agent) or other oligonucleotide-based compounds to cells expressing integrin αvβ6, thereby promoting the uptake of the cargo molecule by these cells.

Description

Integrin ligands and uses thereof

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application is based on, and claims priority to, CN application number 202410233152.X, filed on March 1, 2024, CN application number 202410814083.1, filed on June 21, 2024, CN application number 202411244143.7, filed on September 5, 2024, CN application number 202411378866.6, filed on September 30, 2024, and CN application number 202510198770.X, filed on February 21, 2025. The contents of all these applications are hereby incorporated into this application as a whole. Technical Field

[0003] The present invention relates to the field of biomedicine, and in particular to integrin ligands and uses thereof. Background Art

[0004] Integrin α-vβ-6 (αvβ6), expressed in various cell types, including epithelial cells, is a receptor for the delayed-associated peptide (LAP) of TGF-β and the extracellular matrix (ECM) proteins fibronectin, vitronectin, and tenascin. Although barely detectable in normal healthy adult epithelial cells, αvβ6 integrin is upregulated during wound healing and in various cancers (e.g., colon, ovarian, endometrial, and gastric cancers) and is often associated with poor cancer prognosis. αvβ6 integrin has been shown to promote cell invasion and migration in metastasis and inhibit apoptosis. αvβ6 integrin also regulates the expression of matrix metalloproteinases (MMPs) and activates TGF-β1. Growing evidence, primarily from in vitro studies, suggests that αvβ6 integrin can promote cancer progression. Therefore, integrin αvβ6 is attractive as a tumor marker and potential therapeutic target, particularly given its role in the expression of matrix metalloproteinases (MMPs) and the activation of TGF-β1.

[0005] The in vivo delivery of therapeutically effective compounds, such as drug compounds, to desired cells and / or tissues has been a common challenge for the development of pharmaceutical products. There continues to be a need for stable and effective targeting ligands capable of selectively targeting cells or tissues that can be used to facilitate the targeted delivery of transported / delivered cargo molecules (e.g., therapeutically active compounds or ingredients) to specific cells or tissues. Indeed, there is a general need for targeting ligands that can be conjugated to one or more selected transported molecules, such as one or more drug products or other payloads, to facilitate the in vivo delivery of the transported molecules to the desired cells or tissues. Furthermore, there is a need for compounds that target integrin αvβ6 that are suitable for conjugation to transported molecules to deliver the transported molecules to cells expressing integrin αvβ6 in vivo. For particular transported molecules, such as therapeutic oligonucleotide-based compounds (e.g., antisense oligonucleotides or RNAi agents), there is a need for targeting ligands capable of targeting integrin αβ, which can be conjugated to the oligonucleotide-based compounds to deliver the therapeutic agent to cells and / or tissues expressing integrin αβ and facilitate entry of the therapeutic agent into the cell by receptor-mediated endocytosis, pinocytosis, or by other means. Summary of the Invention

[0006] Described herein is a novel synthetic αvβ6 integrin ligand (also referred to herein as an αvβ6 ligand). The αvβ6 integrin ligand disclosed herein is stable in serum and has affinity for and can specifically bind to the αvβ6 integrin. The αvβ6 integrin ligand can be conjugated to a transported / delivered molecule to facilitate delivery of the transported / delivered molecule to a desired cell or tissue expressing the αvβ6 integrin, such as epithelial cells.

[0007] To this end, in a first aspect of the present invention, the present invention provides a compound of Formula I or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and further provides an αvβ6 integrin ligand, which comprises a compound of Formula I or a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0008] in:

[0009] X is selected from direct bond,

[0010] Preferably, X is selected from a direct bond or

[0011] More preferably, X is a direct bond;

[0012] Y is selected from NH or CH2;

[0013] n is 0, 1 or 2;

[0014] m is 0 or 1;

[0015] Preferably, m is 0;

[0016] R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl;

[0017] R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 alkyl;

[0018] Ring B is selected from 3-10 membered cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aromatic ring, 5-10 membered aromatic heterocycle;

[0019] R 4 Selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 alkoxy, optionally substituted amino;

[0020] R 5a 、R 5b are independently selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 alkoxy, optionally substituted amino;

[0021] Preferably, R 5a 、R 5b are each independently selected from hydrogen, halogen, -C 1～6 alkyl;

[0022] More preferably, R 5a 、R 5b are each independently selected from hydrogen, -C 1～6 alkyl;

[0023] Most preferably, R 5a 、R 5b are each independently selected from hydrogen, methyl;

[0024] Or, R 4 and R 5a Directly connected to form a C containing 1-3 heteroatoms 3～6 Alkylene chains (preferably forming );

[0025] Ring A is selected from 5-10 membered aromatic heterocycles and 3-10 membered heterocycloalkyls, wherein the aromatic heterocycles and heterocycloalkyls are optionally replaced by one, two or three R A replace;

[0026] Each R A are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl);

[0027] Q is selected from -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C(O)NR Q4 -、-C 0～4 Alkylene-(3-10 membered cycloalkyl)-C 0～4 Alkylene-NR Q4 -、-C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C 0～4 Alkylene, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C(O)NR Q4 -、-C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-NR Q4 -、-C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-O-, -C 0～4 Alkylene-CR Q1 R Q2 -NR Q4 -、-C 0～4 Alkylene-(5-12 membered spiroheterocyclic)-, -C 0～4 Alkylene-(5-12 membered fused heterocyclic ring)-, -C 0～4 Alkylene-(5-10 membered aromatic heterocycle)-, C 0～4 Alkylene-(5-10 membered aromatic ring)-NR Q4 -、 The cycloalkyl, heterocycloalkyl, spiro heterocycle, fused heterocycle, aromatic heterocycle, aromatic ring are optionally substituted by one, two or three R Q3 replace;

[0028] Each RQ3 are independently selected from hydrogen, halogen, cyano, =O, nitro, -OH, -C 1～6 Alkyl, halogen-substituted -C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0029] Each R Q4 are independently selected from hydrogen, -C 1～6 Alkyl, halogen-substituted -C 1～6 alkyl;

[0030] Preferably, each R Q4 are independently selected from hydrogen, -C 1～6 alkyl;

[0031] More preferably, each R Q4 are independently selected from hydrogen, -C 1～3 alkyl;

[0032] Most preferably, each R Q4 are independently selected from hydrogen, methyl;

[0033] n2 is an integer from 0 to 6;

[0034] The R Q1 、R Q2 The atoms directly connected to it form a 3-10 membered heterocyclic ring;

[0035] The L1 is selected from

[0036] Preferably, the L1 is selected from

[0037] When X is selected from a direct bond, Y is selected from NH, and Q is selected from R 1 、R 2 When selected from hydrogen, the L1 is not selected from

[0038] In some embodiments, X is selected from a direct bond,

[0039] Preferably, X is selected from a direct bond or

[0040] More preferably, X is a direct bond;

[0041] Y is selected from NH or CH2;

[0042] n is 0, 1 or 2;

[0043] m is 0 or 1;

[0044] Preferably, m is 0;

[0045] R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl;

[0046] R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 alkyl;

[0047] Ring B is selected from 3-10 membered cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aromatic ring, 5-10 membered aromatic heterocycle;

[0048] R 4 Selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 alkoxy, optionally substituted amino;

[0049] R 5a 、R 5b are independently selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 alkoxy, optionally substituted amino;

[0050] Preferably, R 5a 、R 5b are each independently selected from hydrogen, halogen, -C 1～6 alkyl;

[0051] More preferably, R 5a 、R 5b are each independently selected from hydrogen, -C 1～6 alkyl;

[0052] Most preferably, R 5a 、R 5b are each independently selected from hydrogen, methyl;

[0053] Or, R 4 and R 5a Directly connected to form a C containing 1-3 heteroatoms 3～6 Alkylene chains (preferably forming );

[0054] Ring A is selected from 5-10 membered aromatic heterocycles and 3-10 membered heterocycloalkyls, wherein the aromatic heterocycles and heterocycloalkyls are optionally replaced by one, two or three R A replace;

[0055] Each R A are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl);

[0056] Q is selected from -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C 0～4 Alkylene-NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C 0～4 Alkylene, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-O-, -C 0～4 Alkylene-CR Q1 R Q2 -NH-, -C 0～4 Alkylene-(5-12 membered spiroheterocyclic)-, -C 0～4 Alkylene-(5-12 membered fused heterocyclic ring)-, -C 0～4 Alkylene-(5-10 membered aromatic heterocycle)-, The cycloalkyl, heterocycloalkyl, spiro heterocycle, fused heterocycle, aromatic heterocycle are optionally substituted by one, two or three R Q3 replace;

[0057] Each R Q3 are independently selected from hydrogen, halogen, cyano, =O, nitro, -OH, -C 1～6 Alkyl, halogen-substituted -C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0058] n2 is an integer from 0 to 6;

[0059] The R Q1 、R Q2 The atoms directly connected to it form a 3-10 membered heterocyclic ring;

[0060] The L1 is selected from

[0061] Preferably, the L1 is selected from

[0062] When X is selected from a direct bond, Y is selected from NH, and Q is selected from R 1 、R 2 When selected from hydrogen, the L1 is not selected from

[0063] In some embodiments, L1 is selected from The aa end is connected to the B ring.

[0064] In some embodiments, ring A is selected from 5-membered aromatic heterocycle, 6-membered aromatic heterocycle, 9-membered aromatic heterocycle, 5-membered heterocycloalkyl, 6-membered heterocycloalkyl, 9-membered heterocycloalkyl, 10-membered heterocycloalkyl; the aromatic heterocycle and heterocycloalkyl are optionally replaced by one, two or three R A replace.

[0065] In some embodiments, Ring A is selected from

[0066] In some embodiments, each R A are independently selected from hydrogen, =O, -C 1～3 Alkyl, -O(C 1～6 Alkyl), halogen-substituted C 1～3 Alkyl, halogen, -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl).

[0067] In some embodiments, each R A Each is independently selected from hydrogen, =0, methyl, isopropyl, methoxy, trifluoromethyl, fluorine, -NH2, and -NH(CH3).

[0068] In some embodiments, Q is selected from -(7-membered spiroheterocycle)-, -(8-membered spiroheterocycle)-, -(9-membered spiroheterocycle)-, -(10-membered spiroheterocycle)-, -(8-membered fused heterocycle)-, -C3 alkylene-(5-membered aromatic heterocycle)-, -C1 alkylene-(3-membered cycloalkyl)-C(O)NH-, -(4-membered cycloalkyl)-C1 alkylene-NR Q4 -, -(5-membered cycloalkyl)-C(O)NH-, -(6-membered cycloalkyl)-C(O)NH-, -C2 alkylene-(5-membered cycloalkyl)-NH-, -C1 alkylene-(5-membered cycloalkyl)-NH-, -C1 alkylene-(5-membered cycloalkyl)-C1 alkylene-NH-, -C1 alkylene-(5-membered heterocycloalkyl)-, -C2 alkylene-(4-membered heterocycloalkyl)-, -C2 alkylene-(5-membered heterocycloalkyl)-, -C3 alkylene -(5-membered heterocycloalkyl)-, -C1 alkylene-(6-membered heterocycloalkyl)-, -C1 alkylene-(5-membered heterocycloalkyl)-NH-, -C1 alkylene-(5-membered heterocycloalkyl)-O-, -(5-membered heterocycloalkyl)-C(O)NH-, -(6-membered heterocycloalkyl)-C(O)NH-, -C1 alkylene-(5-membered heterocycloalkyl)-C1 alkylene, -C1 alkylene-(6-membered heterocycloalkyl)-C1 alkylene, -C3 alkylene-CR Q1 R Q2 -NH-, -C1 alkylene-phenyl ring-NR Q4 -; the cycloalkyl, heterocycloalkyl, spiro heterocycle, fused heterocycle, aromatic heterocycle, benzene ring is optionally replaced by one, two or three R Q3 replace.

[0069] In some embodiments, Q is selected from -(7-membered spiroheterocycle)-, -(8-membered spiroheterocycle)-, -(9-membered spiroheterocycle)-, -(10-membered spiroheterocycle)-, -(8-membered fused heterocycle)-, -C3 alkylene-(5-membered aromatic heterocycle)-, -C1 alkylene-(3-membered cycloalkyl)-C(O)NH-, -(5-membered cycloalkyl)-C(O)NH-, -(6-membered cycloalkyl)-C(O)NH-, -C2 alkylene-(5-membered cycloalkyl)-NH-, -C1 alkylene-(5-membered cycloalkyl)-NH-, -C1 alkylene-(5-membered cycloalkyl)-C1 alkylene- NH-, -C1 alkylene-(5-membered heterocycloalkyl), -C2 alkylene-(5-membered heterocycloalkyl), -C3 alkylene-(5-membered heterocycloalkyl), -C1 alkylene-(6-membered heterocycloalkyl), -C1 alkylene-(5-membered heterocycloalkyl)-NH-, -C1 alkylene-(5-membered heterocycloalkyl)-O-, -(5-membered heterocycloalkyl)-C(O)NH-, -(6-membered heterocycloalkyl)-C(O)NH-, -C1 alkylene-(5-membered heterocycloalkyl)-C1 alkylene, -C1 alkylene-(6-membered heterocycloalkyl)-C1 alkylene, -C3 alkylene-CR Q1 R Q2-NH-; the cycloalkyl, heterocycloalkyl, spiro heterocycle, fused heterocycle, aromatic heterocycle are optionally substituted by one, two or three R Q3 replace.

[0070] In some embodiments, Q is selected from

[0071] In some embodiments, Q is selected from

[0072] In some embodiments, each R Q3 are independently selected from hydrogen, =O, -C 1～3 Alkyl, halogen-substituted -C 1～3 alkyl.

[0073] In some embodiments, each R Q3 are independently selected from hydrogen, =O, -C 1～3 alkyl.

[0074] In some embodiments, each R Q3 are independently selected from hydrogen, =0, and methyl.

[0075] In some embodiments, the R Q1 、R Q2 The atoms directly connected thereto form a 4-membered heterocyclic ring (preferably forming oxetane).

[0076] In some embodiments, Ring B is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, phenyl, naphthyl, pyridyl, pyrimidinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl, thienyl, benzothienyl, thiazolyl, benzothiazolyl, furanyl, oxazolyl, isoxazolyl, benzofuranyl, indolyl, indazolyl, benzimidazolyl, oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, tetrazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, pyrrolidinyl, dioxanyl, or dioxolanyl.

[0077] In some embodiments, Ring B is naphthyl.

[0078] In some embodiments, Ring B is

[0079] In some embodiments, R 1 、R 2 are independently selected from hydrogen and fluorine.

[0080] In some embodiments, R3 Selected from hydrogen, -C 1～3 alkyl.

[0081] In some embodiments, R 3 For hydrogen.

[0082] In some embodiments, the compound has the structure shown in Formula I-1,

[0083] in:

[0084] X is selected from direct bond,

[0085] Preferably, X is selected from a direct bond or

[0086] More preferably, X is a direct bond;

[0087] Y is selected from NH or CH2;

[0088] Preferably, Y is NH;

[0089] n is 0, 1 or 2;

[0090] m is 0 or 1;

[0091] R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl;

[0092] Preferably, R 1 、R 2 are independently selected from hydrogen and halogen;

[0093] More preferably, R 1 、R 2 are independently selected from hydrogen and fluorine;

[0094] Most preferably, R 1 、R 2 is hydrogen;

[0095] R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 alkyl;

[0096] Preferably, R 3 Selected from hydrogen, -C 1～3 alkyl;

[0097] More preferably, R 3 is hydrogen;

[0098] R 5a 、R5b are independently selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 alkoxy, optionally substituted amino;

[0099] Preferably, R 5a 、R 5b are each independently selected from hydrogen, halogen, -C 1～6 alkyl;

[0100] More preferably, R 5a 、R 5b are each independently selected from hydrogen, -C 1～6 alkyl;

[0101] Most preferably, R 5a 、R 5b are each independently selected from hydrogen, methyl;

[0102] R A1 、R A2 、R A3 are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl);

[0103] Preferably, R A1 、R A2 、R A3 are independently selected from hydrogen, =O, -C 1～3 Alkyl, -O(C 1～6 Alkyl), halogen-substituted C 1～3 Alkyl, halogen, -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0104] More preferably, R A1 、R A2 、R A3 are independently selected from hydrogen, -C 1～3 Alkyl, -O(C1～6 Alkyl), halogen-substituted C 1～3 Alkyl, halogen, -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0105] More preferably, R A1 、R A2 、R A3 Each of the following groups is independently selected from hydrogen, methyl, isopropyl, methoxy, trifluoromethyl, fluorine, -NH2, -NH(CH3);

[0106] Most preferably, R A1 、R A2 、R A3 are independently selected from hydrogen, methyl, isopropyl, methoxy, (preferably, R A1 、R A2 is hydrogen, R A3 is selected from hydrogen, methyl, isopropyl, methoxy);

[0107] Q is selected from -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C 0～4 Alkylene-NR Q4 -、-C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C 0～4 Alkylene, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-O-, -C 0～4 Alkylene-(5-12 membered spiroheterocyclic)-, -C 0～4 Alkylene-(5-12 membered fused heterocyclic ring)-, -C 0～4 Alkylene-(5-10 membered aromatic heterocycle)-, C 0～4 Alkylene-(5-10 membered aromatic ring)-NR Q4 -; the cycloalkyl, heterocycloalkyl, spiro heterocycle, fused heterocycle, aromatic heterocycle, aromatic ring are optionally substituted by one, two or three R Q3 replace;

[0108] Preferably, Q is selected from -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C 0～4 Alkylene-NH-, -C 0～4Alkylene-(3-10 membered heterocycloalkyl)-C 0～4 Alkylene, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-O-, -C 0～4 Alkylene-(5-12 membered spiroheterocyclic)-, -C 0～4 Alkylene-(5-12 membered fused heterocyclic ring)-, -C 0～4 Alkylene-(5-10 membered aromatic heterocycle)-; the cycloalkyl, heterocycloalkyl, spiro heterocycle, fused heterocycle, aromatic heterocycle are optionally replaced by one, two or three R Q3 replace;

[0109] More preferably, Q is selected from -(7-membered spiroheterocycle)-, -(8-membered spiroheterocycle)-, -(9-membered spiroheterocycle)-, -(10-membered spiroheterocycle)-, -(8-membered fused heterocycle)-, -C3 alkylene-(5-membered aromatic heterocycle)-, -C1 alkylene-(3-membered cycloalkyl)-C(O)NH-, -(4-membered cycloalkyl)-C1 alkylene-NR Q4 -、-(5-membered cycloalkyl)-C(O)NH-、-(6-membered cycloalkyl)-C(O)NH-、-C1 alkylene-(5-membered cycloalkyl)-NH-、-C1 alkylene-(5-membered cycloalkyl)-C1 alkylene-NH-、-C1 alkylene-(5-membered heterocycloalkyl)-、-C2 alkylene-(4-membered heterocycloalkyl)-、-C2 alkylene-(5-membered heterocycloalkyl)-、-C3 alkylene-(5-membered heterocycloalkyl)-、-C1 alkylene-(6-membered heterocycloalkyl)-、-C1 alkylene-(5-membered heterocycloalkyl)-NH-、-C1 alkylene-(5-membered heterocycloalkyl)-O-、-(6-membered heterocycloalkyl)-C(O)NH-、-C1 alkylene-(6-membered heterocycloalkyl)-C1 alkylene、-C1 alkylene-phenyl ring-NR Q4 -; the cycloalkyl, heterocycloalkyl, spiro heterocycle, fused heterocycle, aromatic heterocycle, benzene ring is optionally replaced by one, two or three R Q3 replace;

[0110] Further preferably, Q is selected from -(7-membered spiroheterocycle)-, -(8-membered spiroheterocycle)-, -(9-membered spiroheterocycle)-, -(10-membered spiroheterocycle)-, -(8-membered fused heterocycle)-, -C3 alkylene-(5-membered aromatic heterocycle)-, -C1 alkylene-(3-membered cycloalkyl)-C(O)NH-, -(5-membered cycloalkyl)-C(O)NH-, -(6-membered cycloalkyl)-C(O)NH-, -C1 alkylene-(5-membered cycloalkyl)-NH-, -C1 alkylene-(5-membered cycloalkyl)-C1 alkylene-NH-, -C1 alkylene-(5-membered heterocycloalkyl), -C2 alkylene-(5-membered heterocycloalkyl), -C3 alkylene-(5-membered heterocycloalkyl), -C1 alkylene-(6-membered heterocycloalkyl), -C1 alkylene-(5-membered heterocycloalkyl)-NH-, -C1 alkylene-(5-membered heterocycloalkyl)-O-, -(6-membered heterocycloalkyl)-C(O)NH-, -C1 alkylene-(6-membered heterocycloalkyl)-C1 alkylene; the cycloalkyl, heterocycloalkyl, spiro heterocycle, fused heterocycle, aromatic heterocycle are optionally substituted by one, two or three R Q3 replace;

[0111] Each R Q3 are independently selected from hydrogen, halogen, cyano, =O, nitro, -OH, -C 1～6 Alkyl, halogen-substituted -C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0112] Preferably, each R Q3 are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted -C 1～6 alkyl;

[0113] More preferably, each R Q3 are independently selected from hydrogen, =O, -C 1～3 Alkyl, halogen-substituted -C 1～3 alkyl;

[0114] Further preferably, each R Q3 are independently selected from hydrogen, =O, -C 1～3 alkyl;

[0115] Most preferably, each R Q3 are independently selected from hydrogen, =O, methyl;

[0116] Each R Q4 are independently selected from hydrogen, -C 1～6Alkyl, halogen-substituted -C 1～6 alkyl;

[0117] Preferably, each R Q4 are independently selected from hydrogen, -C 1～6 alkyl;

[0118] More preferably, each R Q4 Each independently selected from -C 1～3 alkyl;

[0119] Most preferably, R Q4 is methyl;

[0120] More preferably, Q is selected from

[0121] Most preferably, Q is selected from

[0122] In some embodiments, the compound has the structure shown in Formula I-2,

[0123] n is 0, 1 or 2, preferably 1;

[0124] n2 is 0, 1, 2, 3, 4, 5 or 6, preferably 0, 1, 2, more preferably 1;

[0125] R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl;

[0126] Preferably, R 1 、R 2 are independently selected from hydrogen and halogen;

[0127] More preferably, R 1 、R 2 are independently selected from hydrogen and fluorine;

[0128] Most preferably, R 1 、R 2 is hydrogen;

[0129] R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 alkyl;

[0130] Preferably, R 3 Selected from hydrogen, -C 1～3 alkyl;

[0131] More preferably, R 3 is hydrogen;

[0132] Ring A is selected from 5-10 membered aromatic heterocycles and 3-10 membered heterocycloalkyls, wherein the aromatic heterocycles and heterocycloalkyls are optionally replaced by one, two or three R A replace;

[0133] Preferably, the A ring is selected from a 6-membered aromatic heterocycle, a 9-membered aromatic heterocycle, a 9-membered heterocycloalkyl, or a 10-membered heterocycloalkyl; the aromatic heterocycle or heterocycloalkyl is optionally replaced by one, two, or three R A replace;

[0134] Each R A are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl);

[0135] Preferably, each R A are independently selected from =O, -C 1～3 Alkyl, halogen-substituted C 1～3 Alkyl, halogen, -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0136] More preferably, each R A Each independently selected from =O, methyl, trifluoromethyl, fluorine, -NH2, -NH(CH3);

[0137] Preferably, ring A is selected from

[0138] In some embodiments, the compound has the structure shown in Formula I-3,

[0139] in:

[0140] n is 0, 1 or 2, preferably 1;

[0141] n2 is 0, 1, 2, 3, 4, 5 or 6, preferably 1, 2, 3;

[0142] R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl;

[0143] Preferably, R 1 、R 2 are independently selected from hydrogen and halogen;

[0144] More preferably, R 1 、R 2 are independently selected from hydrogen and fluorine;

[0145] R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 alkyl;

[0146] Preferably, R 3 Selected from hydrogen, -C 1～3 alkyl;

[0147] More preferably, R 3 is hydrogen;

[0148] R A1 、R A2 、R A3 are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl);

[0149] Preferably, R A1 、R A2 、R A3 are independently selected from hydrogen, =O, -C 1～3 Alkyl, halogen-substituted C 1～3 Alkyl, halogen, -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0150] More preferably, R A1 、RA2 、R A3 are independently selected from hydrogen, -C 1～3 Alkyl, halogen-substituted C 1～3 Alkyl, halogen, -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0151] More preferably, R A1 、R A2 、R A3 Each independently selected from hydrogen, methyl, trifluoromethyl, fluorine, -NH2, -NH(CH3);

[0152] Most preferably, R A1 、R A2 、R A3 are independently selected from hydrogen, methyl (preferably, R A1 、R A2 is hydrogen, R A3 is methyl);

[0153] L is selected from

[0154] When R 1 、R 2 When it is hydrogen, L is not

[0155] In some embodiments, the compound has the structure shown in Formula I-4,

[0156] in:

[0157] R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl;

[0158] Preferably, R 1 、R 2 are independently selected from hydrogen and halogen;

[0159] More preferably, R 1 、R 2 are independently selected from hydrogen and fluorine;

[0160] Most preferably, R 1 、R 2 is hydrogen;

[0161] R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 alkyl;

[0162] Preferably, R 3 Selected from hydrogen, -C 1～3 alkyl;

[0163] More preferably, R 3 is hydrogen;

[0164] R 5a 、R 5b are each independently selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 alkoxy, optionally substituted amino;

[0165] Preferably, R 5a 、R 5b are each independently selected from hydrogen, halogen, -C 1～6 alkyl;

[0166] More preferably, R 5a 、R 5b are each independently selected from hydrogen, -C 1～6 alkyl;

[0167] Most preferably, R 5a 、R 5b are each independently selected from hydrogen, methyl;

[0168] Ring A is selected from 5-10 membered aromatic heterocycles and 3-10 membered heterocycloalkyls, wherein the aromatic heterocycles and heterocycloalkyls are optionally replaced by one, two or three (preferably one) R A replace;

[0169] Preferably, the A ring is selected from a 6-membered aromatic heterocycle, a 9-membered heterocycloalkyl, or a 10-membered heterocycloalkyl; the aromatic heterocycle or heterocycloalkyl is optionally replaced by one, two, or three (preferably one) R A replace;

[0170] More preferably, ring A is a benzene ring; the benzene ring is optionally replaced by one, two or three (preferably one) R A replace;

[0171] Each R A are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl);

[0172] Preferably, each R A are independently selected from hydrogen, -C 1～3 Alkyl, halogen-substituted C 1～3 Alkyl, -OH, -O(C 1～6 alkyl);

[0173] More preferably, each R A are independently selected from hydrogen, -C 1～3 Alkyl, -O(C 1～6 alkyl);

[0174] Most preferably, each R A are independently selected from hydrogen, methyl, isopropyl, and methoxy;

[0175] More preferably, ring A is selected from

[0176] Most preferably, Ring A is selected from

[0177] L0 is selected from

[0178] n2 is 0, 1, 2, 3, 4, 5 or 6, preferably 1, 2, 3, more preferably 1;

[0179] Q is selected from -C 0～4 Alkylene-(5-12 membered spiroheterocyclic)-, -C 0～4 Alkylene-(5-12 membered fused heterocycle)-; the spiro heterocycle and fused heterocycle are optionally replaced by one, two or three R Q3 replace;

[0180] Preferably, Q is selected from -(7-membered spiro heterocycle)-, -(8-membered spiro heterocycle)-, -(9-membered spiro heterocycle)-, -(8-membered fused heterocycle)-; the spiro heterocycle and fused heterocycle are optionally replaced by one, two or three R Q3 replace;

[0181] Or preferably, Q is selected from -C 0～4 Alkylene-(5-12 membered spiroheterocycle)-; the spiroheterocycle is optionally replaced by one, two or three R Q3 replace;

[0182] Or preferably, Q is selected from -(7-membered spiro heterocycle)-, -(8-membered spiro heterocycle)-, -(9-membered spiro heterocycle)-; the spiro heterocycle is optionally replaced by one, two or three RQ3 replace;

[0183] Each R Q3 are independently selected from hydrogen, halogen, cyano, =O, nitro, -OH, -C 1～6 Alkyl, halogen-substituted -C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0184] Preferably, each R Q3 are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted -C 1～6 alkyl;

[0185] More preferably, each R Q3 are independently selected from hydrogen, =O, -C 1～3 Alkyl, halogen-substituted -C 1～3 alkyl;

[0186] Further preferably, each R Q3 are independently selected from hydrogen, =O, -C 1～3 alkyl;

[0187] More preferably, each R Q3 are independently selected from hydrogen, =O;

[0188] Most preferably, each R Q3 is hydrogen;

[0189] More preferably, Q is selected from

[0190] More preferably, Q is selected from

[0191] Most preferably, Q is selected from

[0192] Preferably, L0 is selected from

[0193] More preferably, L0 is selected from

[0194] Most preferably, L0 is selected from

[0195] In some embodiments, the compound is selected from:

[0196] Further selected from:

[0197] In a second aspect, the present invention provides an αvβ6 integrin ligand comprising a targeting portion, wherein the targeting portion comprises the compound described in any technical solution of the first aspect, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or comprises a group formed by removing one or more (preferably one) hydrogen atoms from the compound, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0198] In some embodiments, the compound has the structure shown in Formula I,

[0199] X, Y, n, m, R 1 、R 2 、R 3 、R 4 、R 5a 、R 5b , Ring A, Ring B, Q, and L1 are each independently as described in any technical solution of the first aspect;

[0200] Wherein, when the targeting portion comprises a group formed by removing one or more (preferably one) hydrogen atoms from the compound or its stereoisomer or its pharmaceutically acceptable salt, in R 3 、R 4 or removing one or more (preferably one) hydrogen atoms from the B ring (preferably from the B ring);

[0201] Preferably, Ring B is When the targeting moiety comprises a group represented by Formula Ia, X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b , Ring A, Q, and L1 are each independently as described in any technical solution of the first aspect,

[0202] In some embodiments, the compound has the structure shown in Formula I-1,

[0203] X, Y, n, m, R 1 、R 2、R 3 、R 5a 、R 5b 、R A1 、R A2 、R A3 , Q are each independently as described in any technical solution of the first aspect,

[0204] Wherein, when the targeting portion comprises a group formed by removing one or more (preferably one) hydrogen atoms from the compound or its stereoisomer or its pharmaceutically acceptable salt, in R 3 or Above (preferably ) removing one or more (preferably one) hydrogen atoms;

[0205] Preferably, the targeting moiety comprises a group represented by formula I-1a, wherein X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b 、R A1 、R A2 、R A3 , Q are each independently as described in any technical solution of the first aspect,

[0206] In some embodiments, the compound has the structure shown in Formula I-2,

[0207] n、n2、R 1 、R 2 、R 3 , Ring A are each independently as described in any technical solution of the first aspect,

[0208] Wherein, when the targeting portion comprises a group formed by removing one or more (preferably one) hydrogen atoms from the compound or its stereoisomer or its pharmaceutically acceptable salt, in R 3 or Above (preferably ) removing one or more (preferably one) hydrogen atoms;

[0209] Preferably, the targeting moiety comprises a group represented by formula I-2a, wherein n, n2, R 1 、R 2 、R 3 , Ring A are each independently as described in any technical solution of the first aspect,

[0210] In some embodiments, the compound has the structure shown in Formula I-3,

[0211] n、n2、R 1 、R 2 、R 3 、R A1 、R A2 、R A3 , L are each independently as described in any technical solution of the first aspect,

[0212] Wherein, when the targeting portion comprises a group formed by removing one or more (preferably one) hydrogen atoms from the compound or its stereoisomer or its pharmaceutically acceptable salt, in R 3 or Above (preferably ) removing one or more (preferably one) hydrogen atoms;

[0213] Preferably, the targeting moiety comprises a group shown in formula I-3a, wherein n, n2, R 1 、R 2 、R 3 、R A1 、R A2 、R A3 , L are each independently as described in any technical solution of the first aspect,

[0214] In some embodiments, the compound has the structure shown in Formula I-4,

[0215] R 1 、R 2 、R 3 、R 5a 、R 5b , L0, and A ring are each independently as described in any technical solution of the first aspect,

[0216] Wherein, when the targeting portion comprises a group formed by removing one or more (preferably one) hydrogen atoms from the compound or its stereoisomer or its pharmaceutically acceptable salt, in R 3 or Above (preferably ) removing one or more (preferably one) hydrogen atoms;

[0217] Preferably, the targeting moiety comprises a group represented by formula I-4a, R 1 、R 2 、R 3 、R 5a 、R 5b , L0, and A ring are each independently as described in any technical solution of the first aspect,

[0218] In some embodiments, the targeting moiety comprises the compound described in the first aspect or comprises a group selected from the group consisting of:

[0219] In some embodiments, the αvβ6 integrin ligand further comprises a linker moiety that is covalently attached to the targeting moiety.

[0220] In some embodiments, the linker moiety comprises a PEG linker.

[0221] In some embodiments, the linker moiety is n1 is selected from integers of 2 to 20, preferably from integers of 2 to 10.

[0222] In some embodiments, the targeting moiety comprises a group formed by removing a hydrogen atom from a compound of Formula I or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and the linker moiety is The linker moiety is covalently linked to the targeting moiety,

[0223] X, Y, n, m, R 1 、R 2 、R 3 、R 4 、R 5a 、R 5b , Ring A, Ring B, Q, and L1 are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10;

[0224] Wherein, the targeting portion comprises a compound of formula I or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in R 3 、R 4 or a group formed by removing a hydrogen atom from the B ring (preferably from the B ring);

[0225] Preferably, Ring B is When the αvβ6 integrin ligand comprises a group represented by formula Ib, X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b , Ring A, Q, and L1 are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10,

[0226] In some embodiments, the targeting moiety comprises a group formed by removing a hydrogen atom from a compound of formula I-1 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and the linker moiety is The linker moiety is covalently linked to the targeting moiety,

[0227] X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b 、R A1 、R A2 、R A3 , Q are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10;

[0228] Wherein, the targeting portion comprises a compound represented by formula I-1 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in R 3 or Above (preferably A group formed by removing a hydrogen atom;

[0229] Preferably, the αvβ6 integrin ligand comprises a group represented by formula I-1b, wherein X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b 、R A1 、R A2 、R A3 , Q are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10,

[0230] In some embodiments, the targeting moiety comprises a group formed by removing a hydrogen atom from a compound of formula I-2 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and the linker moiety is The linker moiety is covalently linked to the targeting moiety,

[0231] n、n2、R 1 、R 2 、R 3 , Ring A are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10;

[0232] Wherein, the targeting portion comprises a compound represented by formula I-2 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in R 3 or Above (preferably A group formed by removing a hydrogen atom;

[0233] Preferably, the αvβ6 integrin ligand comprises a group shown in formula I-2b, wherein n, n2, R 1 、R 2 、R 3 , Ring A are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10,

[0234] In some embodiments, the targeting moiety comprises a group formed by removing a hydrogen atom from a compound of formula I-3 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and the linker moiety is The linker moiety is covalently linked to the targeting moiety,

[0235] n、n2、R 1 、R 2 、R 3 、R A1 、R A2 、R A3 , L are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10;

[0236] Wherein, the targeting portion comprises a compound represented by formula I-3 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in R 3 or Above (preferably A group formed by removing a hydrogen atom;

[0237] Preferably, the αvβ6 integrin ligand comprises a group shown in formula I-3b, wherein n, n2, R 1 、R 2 、R 3 、R A1 、R A2 、R A3 , L are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10,

[0238] In some embodiments, the targeting moiety comprises a group formed by removing a hydrogen atom from a compound of formula I-4 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and the linker moiety is The linker moiety is covalently linked to the targeting moiety,

[0239] R 1 、R 2 、R 3 、R 5a 、R 5b , L0, and A ring are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10;

[0240] Wherein, the targeting portion comprises a compound represented by formula I-4 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in R 3 or Above (preferably A group formed by removing a hydrogen atom;

[0241] Preferably, the αvβ6 integrin ligand comprises a group represented by formula I-4b, R 1 、R 2 、R 3 、R 5a 、R 5b , L0, and A ring are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10,

[0242] In some embodiments, the αvβ6 integrin ligand comprises a group selected from:

[0243] In a third aspect, the present invention provides a delivery conjugate targeting αvβ6 integrin, comprising: an αvβ6 integrin ligand, and a group comprising a molecule to be delivered, wherein the αvβ6 integrin ligand and the group comprising the molecule to be delivered are covalently linked, wherein the αvβ6 integrin ligand is as described in any one of the technical solutions of the first or second aspect.

[0244] In some embodiments, the αvβ6 integrin ligand comprises a targeting portion and a linker portion, wherein the targeting portion comprises a group formed by removing a hydrogen atom from a compound of Formula I or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and the linker portion is The linker moiety is covalently linked to the targeting moiety and the group comprising the molecule to be delivered,

[0245] X, Y, n, m, R 1 、R 2 、R 3 、R 4 、R 5a 、R 5b , Ring A, Ring B, Q, and L1 are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10;

[0246] Wherein, the targeting portion comprises a compound of formula I or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in R 3 、R 4 or a group formed by removing a hydrogen atom from the B ring (preferably from the B ring);

[0247] Preferably, Ring B is When the delivery conjugate targeting αvβ6 integrin comprises a compound represented by formula Ic, X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b , Ring A, Q, and L1 are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, and Z is a group containing the molecule to be delivered,

[0248] In some embodiments, the delivery conjugate targeting αvβ6 integrin comprises a compound represented by Formula I-1c, wherein X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b 、R A1 、R A2 、R A3 , Q are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, Z is a group containing the molecule to be delivered,

[0249] In some embodiments, the delivery conjugate targeting αvβ6 integrin comprises a compound represented by Formula I-2c, wherein n, n2, R 1 、R 2 、R 3, Ring A are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, Z is a group containing the molecule to be delivered,

[0250] In some embodiments, the delivery conjugate targeting αvβ6 integrin comprises a compound shown in Formula I-3c, wherein n, n2, R 1 、R 2 、R 3 、R A1 、R A2 、R A3 , L are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, Z is a group containing the molecule to be delivered,

[0251] In some embodiments, the delivery conjugate targeting αvβ6 integrin comprises a compound shown in Formula I-4c, R 1 、R 2 、R 3 、R 5a 、R 5b , L0, and A ring are each independently as described in any technical solution of the first aspect, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, and Z is a group containing the molecule to be delivered,

[0252] In some embodiments, the delivery conjugate targeting αvβ6 integrin is selected from the group consisting of:

[0253] Each Z independently represents the group comprising the molecule to be delivered.

[0254] In some embodiments, the group comprising the delivered molecule further comprises a linker group that is covalently attached to the αvβ6 integrin ligand and the delivered molecule.

[0255] In some embodiments, the linker group is in:

[0256] L a1 Selected from p1 is selected from 1, 2, 3, 4, 5, preferably 3; preferably, L a1 One side of the carbonyl end is covalently linked to the molecule being delivered, and the other side of the carbonyl end is covalently linked to L a2 covalent attachment;

[0257] L a2 Selected from p2 is selected from 1, 2, 3, 4, 5, preferably 2; preferably, L a2 The amino terminus of L a1 Covalently linked, carbonyl end and L a3 covalent attachment;

[0258] L a3 Selected from Each p3 is independently selected from 1, 2, 3, 4, 5, preferably 2; preferably, L a3 The amino terminus of L a2 Covalently linked, the triazole N-atom end or the triazole C-atom end is covalently linked to the αvβ6 integrin ligand;

[0259] Preferably, L a3 Selected from p3 is selected from 1, 2, 3, 4, 5, preferably 2; preferably, L a3 The amino terminus of L a2 Covalently linked, the triazole N-atom end is covalently linked to the αvβ6 integrin ligand.

[0260] In some embodiments, the linker group is selected from The carbonyl end is covalently linked to the delivered molecule, and the triazole N-atom end or the triazole C-atom end is covalently linked to the αvβ6 integrin ligand.

[0261] In some embodiments, the linker group is The carbonyl end is covalently linked to the delivered molecule, and the triazole N-atom end is covalently linked to the αvβ6 integrin ligand.

[0262] In some embodiments, the delivered molecule includes, but is not limited to, an RNAi agent, a small molecule, an antibody, an antibody fragment, an immunoglobulin, a monoclonal antibody, a label or marker, a lipid, a natural or modified nucleic acid, a natural or modified nucleic acid oligonucleotide, a natural or modified nucleic acid polynucleotide, a peptide, an aptamer, a polymer, a polyamine, a protein, a toxin, a vitamin, polyethylene glycol, a hapten, digoxigenin, biotin, a radioactive atom or molecule, or a fluorophore.

[0263] In some embodiments, the delivered molecule is a RNAi agent.

[0264] In some embodiments, the delivered molecular targets include but are not limited to the following targets: MMP7, RAGE, SOD1.

[0265] In some embodiments, the delivered molecule is a small interfering RNA (siRNA), the siRNA comprising a sense strand and an antisense strand, and the sequences of the sense strand and antisense strand of the siRNA are selected from the sense strand and antisense strand sequences of any duplex 1 to duplex 3 described in Table 6.

[0266] In some embodiments, the delivery conjugate targeting αvβ6 integrin is selected from the group consisting of:

[0267] Each Z R independently represent the molecules to be delivered, preferably, each Z R RNAi agents are represented independently.

[0268] In a fourth aspect, the present invention provides an αvβ6 integrin ligand precursor comprising a targeting moiety and a reactive group for conjugating with a group comprising a molecule to be delivered, wherein the targeting moiety is as described in any technical solution of the second aspect.

[0269] In some embodiments, the reactive group is selected from azide and alkyne.

[0270] In some embodiments, the αvβ6 integrin ligand precursor further comprises a linker moiety that is covalently linked to the targeting moiety and the reactive group.

[0271] In some embodiments, the linker moiety comprises a PEG linker.

[0272] In some embodiments, the linker moiety is n1 is selected from integers of 2 to 20, preferably from integers of 2 to 10.

[0273] In some embodiments, the present invention provides a compound of formula Id or a stereoisomer thereof or a pharmaceutically acceptable salt thereof. The present invention also provides an αvβ6 integrin ligand precursor, which is a compound of formula Id or a stereoisomer thereof or a pharmaceutically acceptable salt thereof.

[0274] in:

[0275] X, Y, n, m, R 1 、R 2 、R 3 、R4 、R 5a 、R 5b , Ring A, Ring B, Q, and L1 are each independently as described in any technical solution of the first aspect;

[0276] L2 is n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10;

[0277] R 6 It is selected from an azido group and an alkynyl group, and is preferably an azido group (-N3).

[0278] In some embodiments, the compound has the structural formula shown in Formula I-1d,

[0279] in:

[0280] X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b 、R A1 、R A2 、R A3 , Q are each independently as described in any technical solution of the first aspect;

[0281] n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10;

[0282] R 6 It is selected from an azido group and an alkynyl group, and is preferably an azido group (-N3).

[0283] In some embodiments, the compound has the structural formula shown in Formula I-2d,

[0284] in:

[0285] n、n2、R 1 、R 2 、R 3 , Ring A are each independently as described in any technical solution of the first aspect;

[0286] n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10;

[0287] R 6 It is selected from an azido group and an alkynyl group, and is preferably an azido group (-N3).

[0288] In some embodiments, the compound has the structural formula shown in Formula I-3d,

[0289] in:

[0290] n、n2、R 1 、R 2 、R 3 、R A1 、R A2 、R A3 , L are each independently as described in any technical solution of the first aspect;

[0291] n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10;

[0292] R 6 It is selected from an azido group and an alkynyl group, and is preferably an azido group (-N3).

[0293] In some embodiments, the compound has the structural formula shown in Formula I-4d,

[0294] in:

[0295] R 1 、R 2 、R 3 、R 5a 、R 5b , L0, and A ring are each independently as described in any technical solution of the first aspect;

[0296] n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10;

[0297] R 6 It is selected from an azido group and an alkynyl group, and is preferably an azido group (-N3).

[0298] In some embodiments, the compound is selected from the group consisting of:

[0299] In a fifth aspect of the present invention, the present invention provides a linker compound, which is a compound represented by Formula IIp or a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0300] in:

[0301] L a1 Selected from p1 is selected from 1, 2, 3, 4, 5, preferably 3; preferably, L a1 The carbonyl end on one side is covalently linked to the hydroxyl group, and the carbonyl end on the other side is covalently linked to L a2 covalent attachment;

[0302] L a2 Selected from p2 is selected from 1, 2, 3, 4, 5, preferably 2; preferably, L a2 The amino terminus of L a1 Covalently linked, carbonyl end and L a3p covalent attachment;

[0303] L a3p Selected from p3 is selected from 1, 2, 3, 4, 5, preferably 2; preferably, L a3p The amino terminus of L a2 Covalently linked.

[0304] In some embodiments, the linker compound is

[0305] In a sixth aspect, the present invention provides a composition or pharmaceutical composition comprising the compound or stereoisomer thereof or a pharmaceutically acceptable salt thereof according to any technical solution of the first aspect, or the αvβ6 integrin ligand according to any technical solution of the second aspect, or the αvβ6 integrin targeted delivery conjugate according to any technical solution of the third aspect, or the αvβ6 integrin ligand precursor according to any technical solution of the fourth aspect, or the linker compound according to any technical solution of the fifth aspect; and optionally, a pharmaceutically acceptable excipient.

[0306] In the seventh aspect of the present invention, the present invention provides use of the compound or stereoisomer thereof or pharmaceutically acceptable salt thereof according to any technical solution of the first aspect, or the αvβ6 integrin ligand according to any technical solution of the second aspect, or the delivery conjugate targeted to αvβ6 integrin according to any technical solution of the third aspect, or the αvβ6 integrin ligand precursor according to any technical solution of the fourth aspect, or the linker compound according to any technical solution of the fifth aspect, or the composition or pharmaceutical composition according to any technical solution of the sixth aspect in the preparation of an agent or drug for delivering a target molecule.

[0307] In an eighth aspect, the present invention provides a method for delivering a target molecule to tissues and / or cells expressing αvβ6 integrin, comprising: administering to a subject the compound according to any technical solution of the first aspect, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the αvβ6 integrin ligand according to any technical solution of the second aspect, or the αvβ6 integrin targeted delivery conjugate according to any technical solution of the third aspect, or the αvβ6 integrin ligand precursor according to any technical solution of the fourth aspect, or the linker compound according to any technical solution of the fifth aspect, or the composition or pharmaceutical composition according to any technical solution of the sixth aspect.

[0308] In the ninth aspect of the present invention, the present invention provides use of the compound of any technical solution of the first aspect, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the αvβ6 integrin ligand of any technical solution of the second aspect, or the delivery conjugate targeted to αvβ6 integrin of any technical solution of the third aspect, or the αvβ6 integrin ligand precursor of any technical solution of the fourth aspect, or the linker compound of any technical solution of the fifth aspect, or the composition or pharmaceutical composition of any technical solution of the sixth aspect in the preparation of a medicament for treating and / or preventing a disease.

[0309] In the tenth aspect of the present invention, the present invention provides a compound or a stereoisomer thereof or a pharmaceutically acceptable salt thereof according to any technical solution of the first aspect, or the αvβ6 integrin ligand according to any technical solution of the second aspect, or the delivery conjugate targeted to αvβ6 integrin according to any technical solution of the third aspect, or the αvβ6 integrin ligand precursor according to any technical solution of the fourth aspect, or the linker compound according to any technical solution of the fifth aspect, or the composition or pharmaceutical composition according to any technical solution of the sixth aspect, for use in treating and / or preventing a disease.

[0310] In the eleventh aspect of the present invention, the present invention provides a method for treating and / or preventing a disease, comprising: administering to a subject an effective amount of the compound of any technical solution of the first aspect, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the αvβ6 integrin ligand of any technical solution of the second aspect, or the delivery conjugate targeted to αvβ6 integrin of any technical solution of the third aspect, or the αvβ6 integrin ligand precursor of any technical solution of the fourth aspect, or the linker compound of any technical solution of the fifth aspect, or the composition or pharmaceutical composition of any technical solution of the sixth aspect.

[0311] In some embodiments, the disease is an epithelial tumor (carcinoma).

[0312] In some embodiments, the disease is idiopathic pulmonary fibrosis, cystic fibrosis, cystic fibrosis with lung cancer, asthma, chronic obstructive pulmonary disease, primary ciliary dyskinesia, non-cystic fibrosis bronchiectasis, pneumonia, chronic bronchitis, respiratory tract infection, interstitial lung disease.

[0313] In some embodiments, the disease is dry eye, periodontal disease, inflammatory bowel disease, atopic dermatitis, or urticaria.

[0314] On the other hand, the present invention also provides the following technical solutions:

[0315] In some embodiments, the αvβ6 integrin ligands disclosed herein have the structure shown in Formula I:

[0316] in,

[0317] Y is selected from NH or CH2; n is 0, 1 or 2;

[0318] R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl;

[0319] R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 the alkyl group or groups comprising the molecule being transported;

[0320] Ring B is selected from 3-10 membered cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aromatic ring, 5-10 membered aromatic heterocycle;

[0321] R 4 Selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 an alkoxy group, an optionally substituted amino group, or a group comprising the molecule being transported;

[0322] Ring A is selected from 5-10 membered aromatic heterocycles and 3-10 membered heterocycloalkyls, wherein the aromatic heterocycles and heterocycloalkyls are optionally replaced by one, two or three R A replace;

[0323] Each R A are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl);

[0324] Q is selected from -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C 0～4 Alkylene-NH-, -C0～4 Alkylene-(3-10 membered heterocycloalkyl)-C 0～4 Alkylene, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-O-, -C 0～4 Alkylene-CR Q1 R Q2 -NH-, -C 0～4 Alkylene-(5-12 membered spiroheterocyclic)-, -C 0～4 Alkylene-(5-10 membered aromatic heterocycle)-, The cycloalkyl, heterocycloalkyl, spiro heterocycle, aromatic heterocycle are optionally substituted by one, two or three R Q3 replace;

[0325] Each R Q3 are independently selected from hydrogen, halogen, cyano, =O, nitro, -OH, -C 1～6 Alkyl, halogen-substituted -C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0326] n2 is an integer from 0 to 6;

[0327] The R Q1 、R Q2 The atoms directly connected to it form a 3-10 membered heterocyclic ring;

[0328] The L1 is selected from

[0329] When Y is selected from NH, Q is selected from R 1 、R 2 When selected from hydrogen, the L1 is not selected from

[0330] Preferably, the L1 is selected from The aa end is connected to the B ring.

[0331] In some embodiments, the αvβ6 integrin ligand disclosed herein has the structure shown in Formula II:

[0332] in,

[0333] Y is selected from NH or CH2; n is 0, 1 or 2;

[0334] R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl;

[0335] R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 the alkyl group or groups comprising the molecule being transported;

[0336] Ring B is selected from 3-10 membered cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aromatic ring, 5-10 membered aromatic heterocycle;

[0337] R 4 Selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 an alkoxy group, an optionally substituted amino group, or a group comprising the molecule being transported;

[0338] Ring A is selected from 5-10 membered aromatic heterocycles and 3-10 membered heterocycloalkyls, wherein the aromatic heterocycles and heterocycloalkyls are optionally replaced by one, two or three R A replace;

[0339] Each R A are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl);

[0340] Q is selected from -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C 0～4 Alkylene-NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C 0～4 Alkylene, -C 0～4Alkylene-(3-10 membered heterocycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-O-, -C 0～4 Alkylene-CR Q1 R Q2 -NH-, -C 0～4 Alkylene-(5-12 membered spiroheterocyclic)-, -C 0～4 Alkylene-(5-10 membered aromatic heterocycle)-, The cycloalkyl, heterocycloalkyl, spiro heterocycle, aromatic heterocycle are optionally substituted by one, two or three R Q3 replace;

[0341] Each R Q3 are independently selected from hydrogen, halogen, cyano, =O, nitro, -OH, -C 1～6 Alkyl, halogen-substituted -C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0342] n2 is an integer from 0 to 6;

[0343] The R Q1 、R Q2 The atoms directly connected to it form a 3-10 membered heterocyclic ring;

[0344] The L is selected from

[0345] When Y is selected from NH, Q is selected from R 1 、R 2 When selected from hydrogen, the L is not selected from

[0346] In some embodiments, the αvβ6 integrin ligand disclosed herein has the structure shown in Formula III:

[0347] in,

[0348] Y is selected from NH or CH2; n is 0, 1 or 2;

[0349] R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6alkyl;

[0350] R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 the alkyl group or groups comprising the molecule being transported;

[0351] R 4 Selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 an alkoxy group, an optionally substituted amino group, or a group comprising the molecule being transported;

[0352] Ring A is selected from 5-10 membered aromatic heterocycles and 3-10 membered heterocycloalkyls, wherein the aromatic heterocycles and heterocycloalkyls are optionally replaced by one, two or three R A replace;

[0353] Each R A are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl);

[0354] Q is selected from -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C 0～4 Alkylene-NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C 0～4 Alkylene, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-O-, -C 0～4 Alkylene-CR Q1 R Q2 -NH-, -C 0～4 Alkylene-(5-12 membered spiroheterocyclic)-, -C 0～4Alkylene-(5-10 membered aromatic heterocycle)-, The cycloalkyl, heterocycloalkyl, spiro heterocycle, aromatic heterocycle are optionally substituted by one, two or three R Q3 replace;

[0355] Each R Q3 are independently selected from hydrogen, halogen, cyano, =O, nitro, -OH, -C 1～6 Alkyl, halogen-substituted -C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0356] n2 is an integer from 0 to 6;

[0357] The R Q1 、R Q2 The atoms directly connected to it form a 3-10 membered heterocyclic ring;

[0358] The L is selected from

[0359] When Y is selected from NH, Q is selected from R 1 、R 2 When selected from hydrogen, the L is not selected from

[0360] Preferably, the A ring is selected from 5-membered aromatic heterocycle, 6-membered aromatic heterocycle, 9-membered aromatic heterocycle, 5-membered heterocycloalkyl, 6-membered heterocycloalkyl, 9-membered heterocycloalkyl, 10-membered heterocycloalkyl; the aromatic heterocycle and heterocycloalkyl are optionally replaced by one, two or three R A replace.

[0361] As a preference: the R A Selected from =O, -C 1～3 Alkyl, halogen-substituted C 1～3 Alkyl, halogen.

[0362] Further: the A ring is selected from

[0363] Preferably, Q is selected from -(7-membered spiroheterocycle)-, -(8-membered spiroheterocycle)-, -(9-membered spiroheterocycle)-, -(10-membered spiroheterocycle)-, -C3 alkylene-(5-membered aromatic heterocycle)-, -C1 alkylene-(3-membered cycloalkyl)-C(O)NH-, -(5-membered cycloalkyl)-C(O)NH-, -(6-membered cycloalkyl)-C(O)NH-, -C2 alkylene-(5-membered cycloalkyl)-NH-, -C1 alkylene-(5-membered cycloalkyl)-C1 alkylene-NH-, -C1 alkylene-(5-membered heterocycle)- -C2 alkylene-(5-membered heterocycloalkyl), -C3 alkylene-(5-membered heterocycloalkyl), -C1 alkylene-(6-membered heterocycloalkyl), -C1 alkylene-(5-membered heterocycloalkyl)-NH-, -C1 alkylene-(5-membered heterocycloalkyl)-O-, -(5-membered heterocycloalkyl)-C(O)NH-, -(6-membered heterocycloalkyl)-C(O)NH-, -C1 alkylene-(5-membered heterocycloalkyl)-C1 alkylene, -C1 alkylene-(6-membered heterocycloalkyl)-C1 alkylene, -C3 alkylene-CR Q1 R Q2 -NH-; the cycloalkyl, heterocycloalkyl, spiro heterocycle, aromatic heterocycle are optionally substituted by one, two or three R Q3 replace;

[0364] Each R Q3 are independently selected from hydrogen, =O, -C 1～3 Alkyl, halogen-substituted -C 1～3 alkyl.

[0365] The R Q1 、R Q2 The atoms directly connected to it form a 4-membered heterocyclic ring.

[0366] Further: said Q is selected from

[0367] As a preference: R 1 、R 2 are independently selected from hydrogen and fluorine.

[0368] As a preference: the R 3 Selected from hydrogen, -C 1～3 alkyl.

[0369] Preferably, the B ring is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyrrole, pyrazole, imidazole, thiophene, benzothiophene, thiazole, benzothiazole, furan, oxazole, isoxazole, benzofuran, indole, indazole, benzimidazole, oxadiazole, 1,2,3-triazole, 1,2,4-triazole, tetrazole, quinolyl, isoquinolyl, quinoxalinyl, tetrahydrofuran, tetrahydropyran, piperidine, pyrrolidine, dioxane or dioxolane.

[0370] Preferably, the group comprising the transported molecule further comprises a polyethylene glycol linker having 2 to 20 ethylene oxide units.

[0371] In some embodiments, the αvβ6 integrin ligands disclosed herein include a structure of the formula: in represents the connection point with the part containing the molecule to be transported, and n1 is an integer from 2 to 20.

[0372] More specifically: The compound described in formula III is selected from: in represents the connection point with the part containing the molecule to be transported, and n1 is an integer from 2 to 20.

[0373] In some embodiments, the αvβ6 integrin ligands disclosed herein are those wherein the transported molecule is an active pharmaceutical ingredient or a prodrug.

[0374] Preferably, in the αvβ6 integrin ligand disclosed herein, the transported molecule comprises a small molecule, an antibody, an antibody fragment, an immunoglobulin, a monoclonal antibody, a label or marker, a lipid, a natural or modified nucleic acid, a natural or modified nucleic acid oligonucleotide, a natural or modified nucleic acid polynucleotide, a peptide, an aptamer, a polymer, a polyamine, a protein, a toxin, a vitamin, polyethylene glycol, a hapten, digoxigenin, biotin, a radioactive atom or molecule, or a fluorophore.

[0375] Further: In the αvβ6 integrin ligand disclosed herein, the transported molecule comprises an RNAi agent.

[0376] In some embodiments, the αvβ6 integrin ligand precursor disclosed herein has the structure shown in Formula IIIb:

[0377] in,

[0378] Y is selected from NH or CH2; n is 0, 1 or 2;

[0379] R1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl;

[0380] R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 The alkyl group may contain a linker group conjugated to a reactive group;

[0381] R 4 Selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 Alkoxy, optionally substituted amino, or may contain a linker group conjugated to a reactive group;

[0382] Ring A is selected from 5-10 membered aromatic heterocycles and 3-10 membered heterocycloalkyls, wherein the aromatic heterocycles and heterocycloalkyls are optionally replaced by one, two or three R A replace;

[0383] Each R A are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl);

[0384] Q is selected from -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C 0～4 Alkylene-NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C 0～4 Alkylene, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-O-, -C 0～4 Alkylene-CRQ1 R Q2 -NH-, -C 0～4 Alkylene-(5-12 membered spiroheterocyclic)-, -C 0～4 Alkylene-(5-10 membered aromatic heterocycle)-, The cycloalkyl, heterocycloalkyl, spiro heterocycle, aromatic heterocycle are optionally substituted by one, two or three R Q3 replace;

[0385] Each R Q3 are independently selected from hydrogen, halogen, cyano, =O, nitro, -OH, -C 1～6 Alkyl, halogen-substituted -C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl);

[0386] n2 is an integer from 0 to 6;

[0387] The R Q1 、R Q2 The atoms directly connected to it form a 3-10 membered heterocyclic ring;

[0388] The L is selected from

[0389] When Y is selected from NH, Q is selected from R 1 、R 2 When selected from hydrogen, the L is not selected from

[0390] where R 3 、R 4 At least one of the comprises a linking group conjugated to a reactive group.

[0391] Preferably, the A ring is selected from 5-membered aromatic heterocycle, 6-membered aromatic heterocycle, 9-membered aromatic heterocycle, 5-membered heterocycloalkyl, 6-membered heterocycloalkyl, 9-membered heterocycloalkyl, 10-membered heterocycloalkyl; the aromatic heterocycle and heterocycloalkyl are optionally replaced by one, two or three R A replace.

[0392] As a preference: the R A Selected from =O, -C 1～3 Alkyl, halogen-substituted C 1～3 Alkyl, halogen.

[0393] Further: the A ring is selected from

[0394] Preferably, Q is selected from -(7-membered spiroheterocycle)-, -(8-membered spiroheterocycle)-, -(9-membered spiroheterocycle)-, -(10-membered spiroheterocycle)-, -C3 alkylene-(5-membered aromatic heterocycle)-, -C1 alkylene-(3-membered cycloalkyl)-C(O)NH-, -(5-membered cycloalkyl)-C(O)NH-, -(6-membered cycloalkyl)-C(O)NH-, -C2 alkylene-(5-membered cycloalkyl)-NH-, -C1 alkylene-(5-membered cycloalkyl)-C1 alkylene-NH-, -C1 alkylene-(5-membered heterocycle)- -C2 alkylene-(5-membered heterocycloalkyl), -C3 alkylene-(5-membered heterocycloalkyl), -C1 alkylene-(6-membered heterocycloalkyl), -C1 alkylene-(5-membered heterocycloalkyl)-NH-, -C1 alkylene-(5-membered heterocycloalkyl)-O-, -(5-membered heterocycloalkyl)-C(O)NH-, -(6-membered heterocycloalkyl)-C(O)NH-, -C1 alkylene-(5-membered heterocycloalkyl)-C1 alkylene, -C1 alkylene-(6-membered heterocycloalkyl)-C1 alkylene, -C3 alkylene-CR Q1 R Q2 -NH-; the cycloalkyl, heterocycloalkyl, spiro heterocycle, aromatic heterocycle are optionally substituted by one, two or three R Q3 replace;

[0395] Each R Q3 are independently selected from hydrogen, =O, -C 1～3 Alkyl, halogen-substituted -C 1～3 alkyl.

[0396] The R Q1 、R Q2 The atoms directly connected to it form a 4-membered heterocyclic ring.

[0397] Further: said Q is selected from

[0398] As a preference: R 1 、R 2 are independently selected from hydrogen and fluorine.

[0399] As a preference: the R 3 Selected from hydrogen, -C 1～3 alkyl.

[0400] Preferably, the linking group is a PEG linker.

[0401] Preferably, the PEG linker comprises 2-20 PEG units.

[0402] Further: wherein the reactive group is azide.

[0403] More specifically: wherein the linking group conjugated to the reactive group has the structure: Where n1 is an integer from 2 to 10, represents the point of attachment to the structure of Formula IIIb.

[0404] Further described herein are compositions comprising αβ integrin ligands. The compositions described herein can be pharmaceutical compositions comprising one or more αβ integrin ligands disclosed herein conjugated to one or more therapeutic substances, such as RNAi agents or other transported molecules.

[0405] Also disclosed herein are methods for in vivo delivery of transported molecules to tissues and / or cells expressing αβ integrin, wherein the methods comprise administering to a subject one or more αβ integrin ligands disclosed herein that have been conjugated to one or more transported molecules. Further disclosed are methods for treating a subject suffering from a disease, symptom, or disorder for which delivery of a therapeutic transported molecule (e.g., an active pharmaceutical ingredient) to cells expressing αβ integrin is therapeutic, wherein the methods comprise administering to the subject one or more αβ integrin ligands disclosed herein that have been conjugated to one or more therapeutic transported molecules.

[0406] In some embodiments, described herein are methods of inhibiting the expression of a target gene in a cell, wherein the method comprises administering to the cell an effective amount of one or more αβ integrin ligands that have been conjugated to one or more oligonucleotide-based compounds (e.g., oligonucleotide-based therapeutic agents) capable of inhibiting the expression of the target gene in the cell, such as RNAi agents. In some embodiments, described herein are methods of inhibiting the expression of a target gene in a cell of a subject, wherein the subject is administered an effective amount of one or more αβ integrin ligands that have been conjugated to one or more oligonucleotide-based compounds (e.g., RNAi agents) capable of inhibiting the expression of the target gene in the cell.

[0407] In some embodiments, described herein are methods of treating a subject having a disease or disorder mediated at least in part by target gene expression, wherein the method comprises administering to a subject in need thereof an effective amount of a pharmaceutical composition, wherein the pharmaceutical composition comprises one or more αβ integrin ligands disclosed herein conjugated to one or more oligonucleotide-based compounds (such as RNAi agents).

[0408] The cells are selected from type I and type II alveolar epithelial cells, goblet cells, secretory epithelial cells, ciliated epithelial cells, corneal and conjunctival epithelial cells, dermal epithelial cells, bile duct epithelial cells, intestinal epithelial cells, ductal epithelial cells, glandular epithelial cells, epithelial tumors (cancers), macrophages, endothelial cells, fibroblasts, smooth muscle cells, granulocytes, T cells, and immune cells.

[0409] The compounds and derivatives provided herein can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

[0410] Definitions of terms used in the present invention: Unless otherwise stated, the initial definitions provided for groups or terms in this document apply to the groups or terms throughout the specification; for terms that are not specifically defined herein, they should be given the meaning that a person skilled in the art would give them based on the disclosure and context.

[0411] "Substitution" means that the hydrogen atoms in a molecule are replaced by other different atoms or groups. Or the lone pair of electrons in an atom in a molecule is replaced by other atoms or groups. For example, the lone pair of electrons on the S atom can be replaced by an O atom to form

[0412] "Optionally substituted" means that "substitution" may but need not occur, and the description includes instances where it occurs and instances where it does not occur.

[0413] A "direct bond" refers to a group on the left and right sides being directly connected. For example, in the compound shown in Formula I, if X is a direct bond, the structural formula of the compound shown in Formula I will become Other similar definitions can be understood by referring to the above content.

[0414] The minimum and maximum carbon atom content in a hydrocarbon group is indicated by a prefix, for example, the prefix C a～b Alkyl refers to any alkyl group containing from "a" to "b" carbon atoms. Thus, for example, "C 1～4 "Alkyl" refers to an alkyl group containing 1 to 4 carbon atoms. Of course, those skilled in the art will understand that C0 alkylene means that the group does not exist. For example, "-C0 alkylene-(3-10 membered cycloalkyl)-C(O)NH-" has the same meaning as "-(3-10 membered cycloalkyl)-C(O)NH-". Other similar definitions can be understood with reference to the above content.

[0415] "Alkyl" refers to a saturated hydrocarbon chain having a specified number of member atoms. For example, C1-6 alkyl refers to an alkyl group having 1 to 6 member atoms, such as 1 to 4 member atoms, such as 1 to 3 member atoms. Alkyl groups can be straight or branched. Representative branched alkyl groups have one, two or three branches. Alkyl groups may optionally be substituted with one or more substituents as defined herein. Alkyl groups include methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl and tert-butyl), pentyl (n-pentyl, isopentyl and neopentyl) and hexyl. Alkyl groups can also be part of other groups, such as C1-6 alkoxy.

[0416] "Alkylene" refers to a divalent saturated aliphatic hydrocarbon radical having the specified number of member atoms. a ~ b Alkylene refers to an alkylene group having a to b carbon atoms. Alkylene groups include branched and straight chain hydrocarbon groups. For example, the term "propylene" can be exemplified by the following structure: Likewise, the term "dimethylbutylene" can be exemplified, for example, by any of the following structures: For example, -C0~4 alkylene can be C0 alkylene, C1 alkylene (for example, -CH2-), C2 alkylene (for example, -CH2CH2-, etc.), C3 alkylene or C4 alkylene; C0 alkylene means that the group here does not exist and is connected in the form of a chemical bond, such as A-C0 alkylene-B means AB, that is, the A group and the B group are directly connected by a chemical bond.

[0417] "Alkenyl" refers to a straight or branched chain hydrocarbon group having the specified number of carbon atoms and in some embodiments 2 to 6 carbon atoms or 2 to 4 carbon atoms and having at least one site of vinyl unsaturation (>C=C<). For example, C a-b Alkenyl refers to an alkenyl group having a to b carbon atoms and is intended to include, for example, ethenyl, propenyl, isopropenyl, 1,3-butadienyl, and the like.

[0418] "Alkynyl" refers to a straight or branched monovalent hydrocarbon radical containing at least one triple bond. The term "alkynyl" is also intended to include those hydrocarbon groups having one triple bond and one double bond. For example, (C2-C6)alkynyl is intended to include ethynyl, propynyl, and the like.

[0419] "Halogen" means fluorine, chlorine, bromine or iodine;

[0420] "Oxo" refers to =0, where an oxygen atom simultaneously replaces two hydrogen atoms through a double bond.

[0421] "Halogen-substituted alkyl" means that the hydrogen atoms in the alkyl group may be substituted by one or more halogen atoms. 1～4The halogenalkyl group refers to an alkyl group containing 1 to 4 carbon atoms in which hydrogen atoms are substituted by one or more halogen atoms.

[0422] The "-OR", "-NRR" and the like described in the present invention means that the R group is connected to the oxygen atom or nitrogen atom via a single bond.

[0423] The oxygen atom in "-C(O)R", "-S(O)2R" and the like described in the present invention is connected to the carbon atom or sulfur atom by a double bond, and the R group is connected to the oxygen atom or sulfur atom by a single bond.

[0424] "Cycloalkyl" and "cycloalkane" refer to saturated or partially saturated cyclic groups having carbon atoms and no ring heteroatoms and having a single ring or multiple rings (including fused and combined). For polycyclic ring systems with aromatic and non-aromatic rings without ring heteroatoms, the term "cycloalkyl" is applicable when the point of attachment is at a non-aromatic carbon atom (e.g., 5,6,7,8,-tetrahydronaphthalene-5-yl). The term "cycloalkyl" includes cycloalkenyl groups such as cyclohexenyl. Examples of cycloalkyl groups include, for example, adamantyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclooctyl, cyclopentenyl, and cyclohexenyl. Examples of cycloalkyl groups including polybicycloalkyl ring systems are bicyclohexyl, bicyclopentyl, bicyclooctyl, etc. For example

[0425] "Heterocycle", "heterocycloalkyl" and "heterocycloalkane" refer to a saturated ring or non-aromatic unsaturated cyclic group having a single ring or multiple rings (including fused and combined) containing at least one heteroatom; wherein the heteroatom refers to a nitrogen atom, an oxygen atom, or a sulfur atom;

[0426] "Spirocyclyl" and "spirocycle" are used interchangeably and refer to polycyclic saturated rings or non-aromatic unsaturated hydrocarbon rings in which the rings share a single carbon atom (called a spiro atom). For example, "5- to 12-membered spirocyclyl" refers to a spirocycle having 5 to 12 ring atoms. Spirocycles are classified as bispirocycles or polyspirocycles based on the number of rings, with bispirocycles being preferred.

[0427] "Spiroheterocyclyl" and "spiroheterocycle" are used interchangeably and refer to a non-aromatic saturated ring or a non-aromatic unsaturated ring system having two monocyclic rings sharing a common carbon atom, which is composed of carbon atoms and heteroatoms selected from nitrogen, oxygen, sulfur, and phosphorus. For example, "5- to 12-membered spiroheterocycle" refers to a spiroheterocycle having 5 to 12 ring atoms, of which 1, 2, or 3 ring atoms are heteroatoms.

[0428] "Fused heterocyclyl" and "fused heterocycle" are used interchangeably and refer to two or more fused non-aromatic saturated or non-aromatic unsaturated ring systems composed of carbon atoms and heteroatoms selected from nitrogen, oxygen, sulfur, and phosphorus. For example, a "5- to 12-membered fused heterocycle" refers to a fused heterocycle having 5 to 12 ring atoms, of which 1, 2, or 3 are heteroatoms.

[0429] "Aryl" and "aromatic ring" are used interchangeably to refer to an all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group with a conjugated π electron system, such as "C 6-10 The term "aryl" refers to a monocyclic or bicyclic aromatic group having 6 to 10 carbon atoms. Non-limiting examples of aryl include phenyl, naphthyl, and the like.

[0430] As used herein, "aromatic heterocycle" refers to an aromatic unsaturated ring containing at least one heteroatom; a heteroatom includes nitrogen, oxygen, sulfur, and the like. It is typically an aromatic monocyclic or bicyclic hydrocarbon ring containing multiple ring atoms, one or more of which is selected from O, N, and S. Preferably, there are one to three heteroatoms. Examples of heterocyclic aryl groups include pyridyl, indolyl, quinoxalinyl, quinolyl, isoquinolyl, benzothiophenyl, benzofuranyl, benzothiophenyl, benzopyranyl, benzothiapyranyl, furanyl, pyrrolyl, thiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, oxadiazolyl, benzimidazolyl, benzothiazolyl, and benzoxazolyl.

[0431] "Bridged ring or bridged ring group" refers to a saturated or unsaturated cyclic group formed by two or more cyclic structures sharing two non-adjacent atoms. Specific examples include but are not limited to:

[0432] "Bridged heterocyclic group" and "bridged heterocycle" are used interchangeably and refer to a saturated or unsaturated cyclic group formed by two or more cyclic structures sharing two non-adjacent atoms, which is composed of carbon atoms and heteroatoms selected from nitrogen, oxygen, sulfur, and phosphorus. Specific examples include, but are not limited to:

[0433] "Fused" refers to structures in which two or more rings share one or more bonds.

[0434] "Stereoisomers" include enantiomers and diastereomers or mixtures thereof.

[0435] The "deuterated compound" of the present invention refers to a molecule or group in which one or more hydrogen atoms are replaced by deuterium atoms, wherein the proportion of deuterium atoms is greater than the abundance of deuterium in nature.

[0436] In the sequence of the present invention, unless otherwise specified, capital letters C, G, U, A, and T represent the base composition of the nucleotides, which are unmodified nucleotides; lowercase letter m indicates that the nucleotide adjacent to the right of the identifier m is a 2'-methoxy nucleotide; lowercase letter f (or " / f / ") indicates that the nucleotide adjacent to the right of the identifier f is a 2'-fluoro nucleotide; the identifier * indicates that the two nucleotides adjacent to the left and right of the identifier * are connected by a thiophosphate group; EVP indicates that the nucleotide adjacent to the right is a (E)-vinylphosphonate-modified nucleotide; invAb indicates an inverted abasic residue.

[0437] Among them, those skilled in the art will understand that the structure of 2'-fluoronucleotide is as follows:

[0438] The structure of a 2'-methoxy nucleotide is shown below:

[0439] The structural formula of cPrpu is as follows:

[0440] The structural formula of invAb is shown below:

[0441] The structural formula of C6-NH2 is It will be understood by those skilled in the art that the phosphate group is the phosphate group at the 5' end of the sense strand.

[0442] In the present invention, the term "RNAi agent" refers to a substance that inhibits gene expression through the RNA interference (RNAi) mechanism, which may be unmodified or include various modification groups. RNAi agents include siRNA, shRNA (short hairpin RNA) and miRNA (microRNA), etc. Taking siRNA as an example, those skilled in the art will understand that it may be unmodified or modified. For example, siRNA may be a double-stranded RNA molecule that has not undergone any chemical modification, maintaining the natural chemical structure of RNA, or it may be a double-stranded RNA molecule that has undergone chemical modification, such as ribose modification, phosphate backbone modification, base modification, or 3' or 5' end modification. Specifically in the siRNA of the present invention, for example, the C6-NH2 (its structural formula is as described above) at the 5' end of the sense strand of duplex 2 is one of the modification groups of the siRNA, is included in the structure of the siRNA, and is one of the structural units constituting the siRNA.

[0443] The term "pharmaceutically acceptable" means that a carrier, vehicle, diluent, excipient, and / or formed salt is generally chemically or physically compatible with the other ingredients that make up a pharmaceutical dosage form and physiologically compatible with the receptor.

[0444] The terms "salt" and "pharmaceutically acceptable salt" refer to acidic and / or basic salts of the above-mentioned compounds or their stereoisomers, formed with inorganic and / or organic acids and bases, and also include zwitterionic salts (inner salts), and also include quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly during the final isolation and purification of the compound. They can also be obtained by mixing the above-mentioned compound, or its stereoisomer, with a certain amount of acid or base appropriately (e.g., equivalent amounts). These salts may form a precipitate in the solution and be collected by filtration, or be recovered after evaporation of the solvent, or be obtained by freeze-drying after reaction in an aqueous medium. The salts described in the present invention can be hydrochlorides, sulfates, citrates, benzenesulfonates, hydrobromides, hydrofluorides, phosphates, acetates, propionates, succinates, oxalates, malates, succinates, fumarates, maleates, tartrates or trifluoroacetates of the compound.

[0445] In certain embodiments, one or more compounds of the present invention may be used in combination with one another. Compounds of the present invention may also be used in combination with any other active agent to prepare a drug or pharmaceutical composition for regulating cell function or treating a disease. If a group of compounds is used, these compounds may be administered to a subject simultaneously, separately, or sequentially.

[0446] The term "treat" generally refers to obtaining a desired pharmacological and / or physiological effect. As used herein, "treat" encompasses any treatment of a disease in a patient, including: (a) preventing the disease or symptoms from occurring in a patient who is susceptible to the disease or symptoms but has not yet been diagnosed with the disease; (b) suppressing the symptoms of the disease, i.e., arresting its development; or (c) relieving the symptoms of the disease, i.e., causing regression of the disease or symptoms.

[0447] The term "subject" refers to a vertebrate. In certain embodiments, a vertebrate is a mammal. Mammals include, but are not limited to, livestock (such as cattle), pets (such as cats, dogs, and horses), primates, mice, and rats. In certain embodiments, a mammal is a human.

[0448] The term "effective amount" refers to an amount effective to achieve the desired therapeutic effect at the necessary dosage and time. A "therapeutically effective amount" of a substance / molecule of the present invention may vary according to factors such as the disease state, age, sex, and weight of the individual and the ability of the substance / molecule to elicit a desired response in the individual. A therapeutically effective amount also encompasses an amount in which the therapeutically beneficial effects of the substance / molecule outweigh any toxic or deleterious consequences. In the case of cancer, a therapeutically effective amount of a drug may reduce the number of cancer cells; reduce the size of a tumor; inhibit (i.e., slow down to some extent, preferably stop) the infiltration of cancer cells into surrounding organs; inhibit (i.e., slow down to some extent, preferably stop) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more symptoms associated with cancer to some extent.

[0449] Obviously, based on the above contents of the present invention, according to common technical knowledge and customary means in this field, without departing from the above basic technical ideas of the present invention, other various forms of modifications, replacements or changes can be made.

[0450] The following further describes the above content of the present invention in detail through specific embodiments in the form of examples. However, this should not be construed as limiting the scope of the above subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention fall within the scope of the present invention. DETAILED DESCRIPTION

[0451] The structures of the compounds were confirmed by nuclear magnetic resonance (NMR) and mass spectrometry (MS). NMR shifts (δ) are given in units of 10⁻⁶ (ppm). NMR measurements were performed using a Bruker Avance NOE 400 MHz and a Bruker Avance NEO 600 MHz NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d₆), deuterated chloroform (CDCl₃), and deuterated methanol (CD₃OD), with tetramethylsilane (TMS) as the internal standard.

[0452] LC-MS analysis was performed using a Shimadzu LC-MS2020 (ESI) liquid chromatography-mass spectrometer. HPLC analysis was performed using a Shimadzu LC-20A high-pressure liquid chromatograph. MPLC (medium-pressure preparative chromatography) was performed using a Gilson GX-281 reverse-phase preparative chromatograph. Thin-layer chromatography silica gel plates were Yantai Huanghai HSGF254 or Qingdao GF254. The specifications used for thin-layer chromatography separation and purification products were 0.4 mm to 0.5 mm. Column chromatography typically used Yantai Huanghai 200-300 mesh silica gel as the carrier.

[0453] The known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from companies such as Anaiji Chemical, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, and Bailingwei Technology.

[0454] Unless otherwise specified, the reaction was carried out under a nitrogen atmosphere. Unless otherwise specified, the solution in the examples is an aqueous solution. Unless otherwise specified, the reaction temperature was room temperature. Unless otherwise specified, M is moles per liter. The reagents described in the examples are abbreviated as follows:

[0455] DMF: N,N-dimethylformamide; DCM: dichloromethane; DIPEA: N,N-diisopropylethylamine; HATU: 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate;

[0456] Example 1

[0457] Synthesis of compound 1

[0458] Step 1. Synthesis of compound 1-2

[0459] Under an ice bath, DMF (245 mL) was added to a 1-L three-necked flask, followed by Boc-S-3-amino-3-(4-bromophenyl)-propionic acid (45.00 g, 131.20 mmol) and stirring to fully dissolve. 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (59.83 g, 157.44 mmol) and N,N-diisopropylethylamine (33.85 g, 262.40 mmol) were added to the above mixture and stirring continued for 10 minutes. Methanol (90 mL) was then slowly added dropwise to the above mixture, and the mixture was allowed to stand at room temperature overnight. LC-MS monitoring confirmed that the starting material had reacted completely. The methanol in the mixture was concentrated, washed with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (ethyl acetate:petroleum ether = 5:1) to afford compound 1-2 (45 g, 95.74% yield). LCMS (E+) m / z: 303.9 [M-56] + .

[0460] Step 2. Synthesis of Compound 1-3

[0461] Under nitrogen protection, to a 1 L three-necked flask was added (S)-3-(4-bromophenyl)-3-((tert-butoxycarbonyl)amino)propionic acid methyl ester (25.00 g, 69.83 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1-naphthol (18.80 g, 69.83 mmol), sodium carbonate (14.80 g, 139.66 mmol), tetrakistriphenylphosphine palladium (807.23 mg, 0.70 mmol) and toluene / water = 4:1 (250 mL). The system was stirred at 80 ° C overnight and monitored by LC-MS. After the reaction, the mixture was cooled to 0°C and the pH was adjusted to 5.0. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (petroleum ether:ethyl acetate = 3:1, dichloromethane) to obtain compound 1-3 (2.00 g, 6.80% yield). LCMS (E+) m / z: 420.2 [MH] + .

[0462] Step 3. Synthesis of Compound 1-4

[0463] Compound 1-3 (2.00 g, 4.75 mmol) was added to a 100 mL three-necked flask and dissolved in DMF (20 mL). Compound 1-3' (2.20 g, 5.23 mmol) and potassium carbonate (1.3 g, 9.50 mmol) were then added. The mixture was stirred at 90°C for 4 hours and monitored by LC-MS. After completion of the reaction, the reaction solution was subjected to MPLC to obtain compound 1-4 (2.80 g, 88.38% yield). LCMS (E+) m / z: 689.2 [M+Na] + .

[0464] Step 4. Synthesis of Compound 1-5

[0465] To a 100 mL single-necked flask, add compound 1-4 (2.80 g, 4.20 mmol), followed by dioxane hydrochloride (20 mL). Stir the mixture at room temperature for 2 hours, monitored by LC-MS. After completion of the reaction, concentrate the mixture to yield crude compound 1-5 (3.00 g).

[0466] Step 5. Synthesis of Compound 1-6

[0467] To a 20 mL single-necked vial, the hydrochloride salt of compound 1-5 (1.50 g, 2.48 mmol), N-Boc-glycine (434.00 mg, 2.48 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1.04 g, 2.73 mmol), and DMF (10 mL) were added. After stirring, N,N-diisopropylethylamine (1.38 g, 9.92 mmol) was added. The reaction was allowed to react at room temperature for 2 hours. LC-MS analysis confirmed the complete disappearance of the starting material. The reaction solution was directly analyzed by MPLC to yield compound 1-6 (1.30 g, 72.40% yield).

[0468] Step 6. Synthesis of Compound 1-7

[0469] Compound 1-6 (1.30 g, 1.80 mmol) and dioxane hydrochloride were added to a 50 mL single-necked vial. The reaction was allowed to proceed at room temperature for 2 hours. LC-MS analysis confirmed the complete disappearance of the starting material. The reaction solution was concentrated to afford compound 1-7 (1.10 g, 92.59% yield).

[0470] Step 7. Synthesis of Compound 1-8

[0471] Compound 1-7 (100 mg, 0.15 mmol) was added to a 20 mL reaction flask and dissolved in DMF (5 mL). 5,6,7,8-tetrahydro-1,8-naphthyridine-2-propionic acid (37.00 mg, 0.18 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (74.10 mg, 0.20 mmol) were then added. After stirring, N,N-diisopropylethylamine (58.00 mg, 0.45 mmol) was added. The system was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, the reaction solution was directly analyzed by MPLC to obtain compound 1-8 (120 mg, 98.64% yield). LCMS (E+) m / z: 812.7 [M+H] + .

[0472] Step 8. Synthesis of Compound 1

[0473] Compound 1-8 (120 mg, 0.14 mmol) was added to a 20 mL reaction flask and dissolved in methanol (5 mL). A 1 N aqueous sodium hydroxide solution (5 mL) was then added. The reaction was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, the reaction mixture was directly analyzed by MPLC to yield compound 1 (100 mg, 98.64% yield).

[0474] LCMS (E+) m / z: 798.4 [M+H] + .

[0475] 1H NMR(600MHz,Chloroform-d)δ10.58(s,1H),9.72(s,1H),8.34(d,J=8.3Hz,1H),7 .88(d,J=8.4Hz,1H),7.45(d,J=7.7Hz,3H),7.43–7.40(m,1H),7.38(d,J=7.8Hz, 2H),7.23(d,J=7.2Hz,2H),6.85(d,J=7.9Hz,1H),6.57(s,1H),6.33(d,J=7.2Hz, 1H),5.31(s,1H),4.41–4.23(m,3H),4.02(t,J=4.9Hz,2H),3.86–3.79(m,3H),3.7 1(dd,J=5.8,3.8Hz,2H),3.70–3.62(m,11H),3.42(d,J=5.0Hz,2H),3.36(t,J=5. 1Hz,2H),3.11(ddd,J=14.2,9.0,5.5Hz,1H),3.04(dt,J=14.9,8.2Hz,1H),2.89(d d,J=15.4,5.1Hz,1H),2.81(dd,J=15.4,5.1Hz,1H),2.76(dd,J=15.8,8.0Hz,1H) ,2.69(t,J=6.4Hz,2H),2.61(ddd,J=14.6,8.8,5.4Hz,1H),1.87(q,J=6.1Hz,2H).

[0476] Example 2

[0477] Synthesis of compound 2

[0478] Step 1. Synthesis of compound 2-2

[0479] Under nitrogen, compound 2-1 (10.00 g, 71.43 mmol) and DMF (350 mL) were added to a 1 L reaction flask, cooled to 0°C, and then sodium hydride (3.10 g, 77.50 mmol) was slowly added and stirring continued for 30 minutes. After the system stopped generating gas, 2-(trimethylsilyl)ethoxymethyl chloride (14.20 g, 85.18 mmol) was added dropwise to the reaction system. The system was stirred at room temperature for 2 hours and monitored by LC-MS. After the reaction was completed, the pH was adjusted with hydrochloric acid (3 M, 50 mL), and then water (500 mL) was added. The mixture was extracted with ethyl acetate, washed with saturated brine, and dried over anhydrous sodium sulfate to obtain crude compound 2-2 (19.00 g, 98.51% yield). LCMS (E+) m / z: 257.0 [M+H] + .

[0480] Step 2. Synthesis of compound 2-3

[0481] Under nitrogen, compound 2-2 (5.00 g, 18.51 mmol), N-bromosuccinimide (3.30 g, 18.51 mmol), 2,2-azobisisobutyronitrile (303.00 mg, 1.85 mmol), and chloroform (50 mL) were added to a 100 mL reaction flask. The system was reacted at 60°C for 5 hours. LC-MS monitoring was used. After completion of the reaction, the mixture was concentrated and analyzed by MPLC to obtain compound 2-3 (1.70 g, 26.17% yield). LCMS (E+) m / z: 336.9 [M+H] + .

[0482] Step 3. Synthesis of Compound 2-4

[0483] Under nitrogen, compound 2-3 (5.50 g, 16.40 mmol) was added to a 100 mL three-necked flask and dissolved in THF (20 mL). The mixture was then placed in dry ice-acetonitrile and cooled for 10 minutes. Isopropylmagnesium chloride (2.0 M, 23.8 mL) was slowly added to the mixture, and the reaction was continued at this temperature for 0.5 hours. The mixture was then brought to -78°C and stirred for 10 minutes. DMF (8.50 g, 116.38 mmol) was then added dropwise, and the reaction was allowed to return to room temperature for 0.5 hours. After completion of the reaction, the pH was adjusted to 5 with hydrochloric acid, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. MPLC yielded compound 2-4 (2.10 g, 45.08% yield). LCMS (E+) m / z: 285.3 [M+H] + .

[0484] Step 4. Synthesis of Compound 2-5

[0485] Under nitrogen, compound 2-4 (2.10 g, 7.39 mmol) was added to a 100 mL reaction flask, dissolved in dichloromethane (50 mL), and then (formylmethylene)triphenylphosphine (2.25 g, 7.39 mmol) was added. The system was allowed to react at room temperature for 2 hours. After completion of the reaction, the reaction solution was analyzed by MPLC to obtain compound 2-5 (2.30 g, crude product). LCMS (E+) m / z: 311.3 [M+H] + .

[0486] Step 5. Synthesis of Compound 2-6

[0487] Under nitrogen protection, compound 2-5 (2.30 g, 7.42 mmol) was added to a 100 mL reaction flask and dissolved in 1,2-dichloroethane (50 mL). 2-amino-4-methylpyridine (1.04 g, 9.65 mmol), sodium acetate borohydride (2.04 g, 9.65 mmol), and 5 drops of acetic acid were then added. The system was reacted at room temperature for 2 hours. After the reaction was completed, the reaction solution was analyzed by MPLC to obtain compound 2-6 (2.0 g, 67.05% yield), as monitored by LC-MS. LCMS (E+) m / z: 403.4 [M+H] + .

[0488] Step 6. Synthesis of Compound 2-7

[0489] Under nitrogen protection, compound 2-6 (100.00 mg, 0.25 mmol) was added to a 20 mL reaction flask and dissolved in methanol (5 mL). 10% platinum dioxide (10.00 mg) was then added, and the atmosphere was replaced with hydrogen three times. The system was allowed to react at room temperature for 2 hours. After completion of the reaction, the reaction solution was analyzed by MPLC to obtain compound 2-7 (100.75 mg, 100% yield), as monitored by LC-MS. LCMS (E+) m / z: 405.1 [M+H] + .

[0490] Step 7. Synthesis of Compound 2-8

[0491] Under nitrogen, compound 2-7 (100.75 mg, 0.25 mmol) was added to a 20 mL reaction flask, dissolved in methanol (5 mL), and then sodium hydroxide solution (1 M, 5 mL) was added. The system was allowed to react at room temperature for 2 hours. After completion of the reaction, the reaction solution was analyzed by MPLC to obtain compound 2-8 (80.00 mg, 80% yield), as monitored by LC-MS. LCMS (E+) m / z: 391.5 [M+H] + .

[0492] Step 8. Synthesis of Compound 2-9

[0493] Under nitrogen, compound 2-8 (80.00 mg, 0.21 mmol) was added to a 20 mL reaction flask and dissolved in DMF (5 mL). Compound 1-5 (118.65 mg, 0.21 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (75.01 mg, 0.21 mmol) were then added. After stirring, N,N-diisopropylethylamine (58.00 mg, 0.45 mmol) was added. The system was stirred at room temperature for 2 hours. The system was allowed to react at room temperature for 2 hours. After completion of the reaction, the reaction solution was analyzed by MPLC to obtain compound 2-9 (80.00 mg, 40.61% yield), as monitored by LC-MS. LCMS (E+) m / z: 939.2 [M+H] + .

[0494] Step 9. Synthesis of Compound 2-10

[0495] Compound 2-9 (80.00 mg, 0.085 mmol) was added to a 10 mL reaction flask, dissolved in tetrahydrofuran (5 mL), and then tetrabutylammonium fluoride (22.23 mg, 0.085 mmol) was added. The system was allowed to react at room temperature for 2 hours. After completion of the reaction, the reaction solution was analyzed by MPLC to obtain compound 2-10 (60.00 mg, 87.65% yield), as monitored by LC-MS. LCMS (E+) m / z: 809.1 [M+H] + .

[0496] Step 10. Synthesis of Compound 2

[0497] Compound 2-10 (60.00 mg, 0.074 mmol) was added to a 10 mL reaction vial and dissolved in methanol (5 mL). Then, sodium hydroxide solution (1 M, 5 mL) was added. The system was allowed to react at room temperature for 2 hours. After completion of the reaction, the reaction mixture was analyzed by MPLC to yield compound 2 (40.00 mg, 68.00% yield), as monitored by LC-MS.

[0498] LCMS (E+) m / z: 796.3 [M+H] + .

[0499] 1H NMR(600MHz,Chloroform-d)δ9.62(s,1H),8.33(d,J=8.3Hz,1H),7.99(s,1H),7.83(d,J=8.4Hz,1H),7.63– 7.31(m,8H),7.24(s,1H),6.84(d,J=7.9Hz,1H),6.32(d,J=24.6Hz,2H),5.86(s,1H),4.33(t,J=5.0Hz,2H) ,4.02(t,J=4.9Hz,2H),3.81(dd,J=5.8,3.8Hz,2H),3.71(dd,J=5.8,3.7Hz,2H),3.70–3.67(m,2H),3.67–3 .60(m,10H),3.35(t,J=5.1Hz,2H),3.14(m,2H),3.00(m,2H),2.79(m,2H),2.24(m,3H),2.08–1.89(m,2H).

[0500] Example 3

[0501] Synthesis of compound 3

[0502] Step 1. Synthesis of compound 3-2

[0503] Synthesis reference of compound 3-1: [Fu-An Kang, Zhihua Sui, A diversity-oriented-synthesis protocol for scaffold discovery based on a general synthetic route to spirocycles, Tetrahedron Letters, 52, 32, 2011, 4204-4206].

[0504] Compound 3-1 (900 mg, 5.89 mmol) was added to a 100 mL three-necked flask and dissolved in DMF (20 mL). NaH (170 mg, 7.00 mmol) was slowly added under an ice bath. After 15 minutes, tert-butyl bromoacetate (1.25 g, 6.48 mmol) was added. The mixture was allowed to stand at room temperature overnight. LC-MS confirmed the completion of the reaction. The mixture was slowly added to water, extracted with ethyl acetate, concentrated, and analyzed by MPLC to obtain compound 3-2 (750 mg). LCMS (E+) m / z: 268.1 [M+H] +

[0505] Step 2. Synthesis of compound 3-3

[0506] Compound 3-2 (750 mg, 2.80 mmol) and 4-methyl-2-aminopyridine (302 mg, 2.80 mmol) were added to a 100 mL single-necked flask and dissolved in MeOH (20 mL). NaCNBH3 (350 mg, 5.60 mmol) was slowly added and stirred at 60°C overnight. LC-MS confirmed the completion of the reaction. Water was slowly added, and the mixture was extracted with ethyl acetate. Compound 3-3 (360 mg) was obtained by MPLC. LCMS (E+) m / z: 360.1 [M+H] +

[0507] Step 3. Synthesis of compound 3-4

[0508] Compound 3-3 (360 mg, 1.00 mmol) was added to a 100 mL single-necked flask, dissolved in DCM (3 mL), and TFA (3 mL) was added. After addition, the mixture was heated to 20°C and stirred for 1 hour, monitored by LC-MS. After completion of the reaction, the mixture was spin-dried to afford compound 3-4 (240 mg, 70% yield). LCMS (E+) m / z: 304.1 [M+H] + .

[0509] Step 4. Synthesis of Compound 3-5

[0510] To a 100 mL three-necked flask, compound 3-4 (96.36 mg, 0.32 mmol) and compound 1-5 (180 mg, 0.32 mmol) were added and dissolved in dry DMF (2 mL). HATU (157.02 mg, 0.41 mmol) and DIPEA (123.16 mg, 0.95 mmol) were then added. After the addition, the reaction was stirred at room temperature overnight and monitored by LC-MS. After completion of the reaction, compound 3-5 (140 mg, 51.73% yield) was obtained directly by MPLC. LCMS (E+) m / z: 852.3 [M+H] + .

[0511] Step 5. Synthesis of compound 3

[0512] Compound 3-5 (150 mg, 0.17 mmol) was added to a 100 mL three-necked flask and dissolved in methanol (2 mL). 1 M sodium hydroxide solution (1 mL) was then added. After complete addition, the reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, compound 3 (142 mg, 96.25% yield) was obtained directly by MPLC.

[0513] LCMS (E+) m / z: 838.4 [M+H] + .

[0514] 1H NMR(400MHz,Chloroform-d)δ8.34(d,J=8.4Hz,1H),7.82(d,J=8.3Hz,1H),7.64(s,1H),7. 41(dq,J=13.6,7.9Hz,6H),7.24(s,1H),6.84(d,J=7.8Hz,1H),6.67–6.41(m,2H),5.48(s, 1H),4.33(t,J=4.8Hz,2H),4.02(t,J=4.9Hz,3H),3.89–3.78(m,2H),3.75–3.54(m,7H),3. 35(t,J=5.1Hz,1H),3.12–2.75(m,2H),2.43–2.04(m,6H),2.01–1.58(m,2H),1.34(s,1H).

[0515] Example 4

[0516] Synthesis of compound 4

[0517] Step 1. Synthesis of compound 4-2

[0518] Compound 4-1 (500 mg, 2.30 mmol) was added to a 100 mL three-necked flask and dissolved in pyridine (5 mL). The reaction system was cooled to 0-5°C and TsCl (475.57 mg, 2.30 mmol) was slowly added. After addition, the mixture was heated to room temperature and stirred for 16 hours, monitored by LC-MS. After completion of the reaction, the reaction solution was directly analyzed by MPLC to obtain compound 4-2 (400 mg, 46.79% yield). LCMS (E+) m / z: 372.3 [M+H] + .

[0519] Step 2. Synthesis of compound 4-3

[0520] Compound 4-2 (369 mg, 1.03 mmol) was added to a 10 mL reaction flask and dissolved in DMF (4 mL). Compound 4-2' (215.00 mg, 1.03 mmol) and cesium carbonate (403.64 mg, 1.24 mmol) were added with stirring. After the addition, the temperature was raised to 80°C and stirred for 3 hours, monitored by LC-MS. After the reaction, the reaction solution was filtered and directly analyzed by MPLC to obtain compound 4-3 (80 mg, 19.02% yield). LCMS (E+) m / z: 408.6 [M+H] + .

[0521] Step 3. Synthesis of compound 4-4

[0522] Compound 4-3 (70 mg, 171.78 μmol) was added to a 4 mL reaction vial and dissolved in DMF (0.5 mL). The reaction system was cooled to 0-5°C, and NaH (20.61 mg, 515.33 μmol, 60% purity) was added portionwise. After the addition, the reaction was stirred at 0-5°C for 30 minutes. Bromoacetic acid (35.80 mg, 257.67 μmol) was then added. After the addition, the temperature was raised to room temperature and stirred for 2 hours, monitored by LC-MS. After the reaction was completed, the reaction solution was added dropwise to saturated ammonium chloride (20 mL), extracted with ethyl acetate (30 mL), washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. After the liquid was concentrated, compound 4-4 (40 mg, 43.77% yield) was obtained by MPLC. LCMS (E+) m / z: 466.5 [M+H] + .

[0523] Step 4. Synthesis of compound 4-5

[0524] To a 4 mL reaction vial, compound 4-4 (40 mg, 109.46 μmol) and compound 1-5 (62.03 mg, 109.46 μmol) were added and dissolved in dry DMF (1 mL). HATU (54.07 mg, 142.30 μmol) and DIPEA (42.44 mg, 328.39 μmol) were then added with stirring. After addition, the reaction was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, compound 4-5 (20 mg, 19.99% yield) was obtained directly by MPLC. LCMS (E+) m / z: 914.6 [M+H] + .

[0525] Step 5. Synthesis of Compound 4-6

[0526] Compound 4-5 (20 mg, 21.88 μmol) was added to a 4 mL reaction vial and dissolved in methanol (1 mL). 1 M sodium hydroxide solution (1 mL) was then added. The reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, compound 4-6 (17 mg, 86.32% yield) was obtained directly by MPLC. LCMS (E+) m / z: 900.5 [M+H] + .

[0527] Step 6. Synthesis of compound 4

[0528] Compound 4-6 (17 mg, 16.76 μmol) was added to a 4 mL reaction vial and dissolved in 4 M dioxane hydrochloride (0.5 mL). Stir the reaction at room temperature for 1 hour, monitoring by LC-MS. After completion of the reaction, compound 4 (11 mg, 82.04% yield) was obtained directly by MPLC.

[0529] LCMS (E+) m / z: 800.7 [M+H] + .

[0530] 1 H NMR(600MHz,Chloroform-d)δ10.23(s,1H),8.83(s,1H),8.30(d,J=8.4Hz,1H),8.18(s,1H),7.75 (d,J=8.5Hz,1H),7.61–7.52(m,1H),7.42–7.31(m,6H),7.16(d,J=7.7Hz,1H),6.77(t,J=4.0Hz,2 H),6.49(d,J=6.3Hz,1H),5.54(s,1H),4.42–4.21(m,4H),4.10–3.91(m,4H),3.72–3.59(m,8H),3 .33(t,J=5.0Hz,2H),3.07(s,1H),2.84(d,J=14.1Hz,1H),2.45(s,1H),2.26(s,3H),1.79(s,1H).

[0531] Example 5

[0532] Synthesis of compound 5

[0533] Step 1. Synthesis of compound 5-2

[0534] Under ice bath, tetrahydrofuran (38 mL) was added to a 250 mL single-necked flask, followed by compound 5-1 (2.50 g, 24.24 mmol) and stirred to fully dissolve. 1 M sodium hydroxide solution (24.50 mL) and Boc2O (6.88 g, 31.52 mmol) were slowly added to the above system in sequence, and the reaction system was returned to room temperature and stirred for 3 hours. The reaction solvent was concentrated, water was added, and the mixture was extracted with methyl tert-butyl ether. The aqueous phase was adjusted to pH 2-3 with 1 M hydrochloric acid, extracted with methyl tert-butyl ether, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain compound 5-2 (3.80 g, 77.12% yield). LCMS (E+) m / z: 226.3 [M+Na] + .

[0535] Step 2. Synthesis of compound 5-3

[0536] Compound 5-2 (140.00 mg, 0.68 mmol), N,N-diisopropylethylamine (356.11 mg, 2.76 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (392.65 mg, 1.03 mmol), and DMF (3 mL) were added to a 20 mL single-necked flask. The system was stirred at room temperature for 10 minutes, and then compound 1-7 (212.00 mg, 0.34 mmol) was added. After the addition, the mixture was allowed to react at room temperature for 2 hours until the starting material disappeared as determined by LC-MS. The reaction solution was directly subjected to MPLC to obtain compound 5-3 (250.00 mg, 91.00% yield). LCMS (E+) m / z: 809.4 [M+H] + .

[0537] Step 3. Synthesis of compound 5-4

[0538] Compound 5-3 (240.00 mg, 296.69 μmol) was added to a 20 mL single-necked flask, followed by dichloromethane (1 mL) and 4 M hydrochloric acid dioxane solution (4 mL). After addition, the reaction was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, the mixture was concentrated to give compound 5-4 (200.00 mg, crude product). LCMS (E+) m / z: 709.3 [M+H] + .

[0539] Step 4. Synthesis of compound 5-5

[0540] Compound 5-4 (200.00 mg, 282.17 mmol) was added to a 20 mL reaction flask and dissolved in tetrahydrofuran (1.5 mL) and water (1.5 mL). Lithium hydroxide monohydrate (71.11 mg, 1.69 mmol) was then added. The reaction was stirred at room temperature for 2 hours and monitored by LC-MS. After the reaction was complete, the reaction solution was directly analyzed by MPLC to obtain compound 5-5 (190.00 mg, 96.92% yield). LCMS (E+) m / z: 695.3 [M+H] + .

[0541] Step 5. Synthesis of compound 5

[0542] To a 20 mL sealed tube, compound 5-5 (190.00 mg, 273.47 μmol), 1-cyanopyrrolidine (52.58 mg, 546.94 μmol), and hexafluoroisopropanol (3 mL) were added. After addition, the tube was heated to 120°C and sealed for 16 hours. The reaction solution was directly analyzed by MPLC to yield compound 5 (15.00 mg, 6.94% yield).

[0543] LCMS (E+) m / z: 791.3 [M+H] + .

[0544] 1 H NMR(600MHz, Methanol-d4)δ8.36(d,J=8.4Hz,1H),7.79(t,J=9.3Hz,1H),7.51–7.34(m,6H),7.28–7.26(m,1H),6 .98(dd,J=8.0,2.5Hz,1H),5.45–5.25(m,1H),4.37–4.35(m,2H),4.15–4.06(m,1H),4.04–4.02(m,2H),4.00–3.86 (m,1H),3.82–3.80(m,2H),3.77–3.68(m,3H),3.66(dd,J=6.1,3.6Hz,2H),3.65–3.56(m,8H),3.36–3.34(m,2H), 3.32–3.26(m,7H),2.99–2.90(m,1H),2.75–2.71(m,2H),2.47–2.45(m,1H),2.29–2.20(m,1H),2.03–1.84(m,3H).

[0545] Example 6

[0546] Synthesis of compound 6

[0547] Step 1. Synthesis of compound 6-2

[0548] To a 250 mL single-necked flask were added compound 6-1 (6.44 g, 34.43 mmol), compound 6-1' (8.44 g, 86.08 mmol), cesium carbonate (3.66 g, 103.30 mmol), CuI (2.62 g, 13.77 mmol), Pd(PPh3)2Cl2 (4.83 g, 6.89 mmol), and DMF (100 mL). The reaction system was purged with nitrogen three times, heated to 130°C, and stirred for 16 hours, monitored by LC-MS. After completion of the reaction, the reaction solution was directly analyzed by MPLC to obtain a crude product, which was then subjected to SFC to obtain compound 6-2 (1.0 g, 14.22% yield). LCMS (E+) m / z: 205.1 [M+H] + .

[0549] Step 2. Synthesis of compound 6-3

[0550] To a 4 mL reaction vial, compound 6-2 (20 mg, 97.93 μmol) and compound 1-7 (61.08 mg, 97.93 μmol) were added and dissolved in dry DMF (1 mL). HATU (48.38 mg, 127.31 μmol) and DIPEA (37.90 mg, 293.79 μmol) were added with stirring. After addition, the reaction was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, compound 6-3 (50 mg, 63.04% yield) was obtained directly by MPLC. LCMS (E+) m / z: 810.7 [M+H] + .

[0551] Step 3. Synthesis of compound 6

[0552] Compound 6-3 (50 mg, 61.74 μmol) was added to a 4 mL reaction vial and dissolved in methanol (1 mL). 1 M sodium hydroxide solution (1 mL) was then added. The reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, MPLC was performed to obtain 6 (35 mg, 65.44% yield).

[0553] LCMS (E+) m / z: 796.7 [M+H] + .

[0554] 1 H NMR(600MHz,Chloroform-d)δ12.55(s,1H),8.30(t,J=12.5Hz,2H),8.00–7.61(m,2H),7.43(t,J=7.6Hz,1H),7.40–7.28(m ,5H),7.17(d,J=7.5Hz,1H),7.00(s,1H),6.85–6.75(m,1H),6.39(s,1H),5.54(d,J=7.3Hz,1H),4.30(d,J=5.5Hz,2H),4.01 (t,J=5.0Hz,2H),3.81(dd,J=5.9,3.7Hz,2H),3.71(dd,J=5.7,3.8Hz,2H),3.67(s,1H),3.66(d,J=5.1Hz,2H),3.63(d,J=5. 0Hz, 4H), 3.35 (t, J = 5.1Hz, 2H), 3.19 (s, 2H), 3.04–2.94 (m, 1H), 2.87 (d, J = 15.5Hz, 1H), 2.74 (s, 2H), 2.57 (d, J = 4.8Hz, 3H).

[0555] Example 7

[0556] Synthesis of compound 7

[0557] Step 1. Synthesis of compound 7-2

[0558] To a 1000 mL single-necked flask at room temperature, add tert-butyl alcohol (574 mL), followed by compound 7-1 (20.70 g, 191.42 mmol), and stir to fully dissolve. Di-tert-butyl dicarbonate (45.95 g, 210.56 mmol) was slowly added to the above system. After addition, the mixture was stirred at room temperature for 16 hours. The reaction solvent was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 8:1) to obtain compound 7-2 (24.10 g, 60.46% yield). LCMS (E+) m / z: 209.1 [M+H] + .

[0559] Step 2. Synthesis of compound 7-3

[0560] Compound 7-2 (2.50 g, 12.0 mmol), methyl 4-bromobutyrate (2.28 g, 12.6 mmol), cesium carbonate (4.69 g, 14.41 mmol), and DMF (48 mL) were added to a 250 mL single-necked flask. The mixture was allowed to react at room temperature for 16 hours until the starting material disappeared as determined by LC-MS. The reaction mixture was quenched with saturated sodium bicarbonate solution, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and the organic layer concentrated. The crude product was purified by MPLC to yield compound 7-3 (3.33 g, 89.96% yield). LCMS (E+) m / z: 309.2 [M+H] + .

[0561] Step 3. Synthesis of compound 7-4

[0562] Compound 7-3 (3.50 g, 11.35 mmol) was added to a 250 mL single-necked flask, followed by ethanol (57 mL) and hydrazine hydrate (11.35 g, 227.00 mmol). After the addition was complete, the reaction system was stirred at 80°C for 16 hours and monitored by LC-MS. After the reaction was complete, the mixture was concentrated to give compound 7-4 (3.40 g, crude product). LCMS (E+) m / z: 309.4 [M+H] + .

[0563] Step 4. Synthesis of compound 7-5

[0564] To a 10 mL reaction vial, compound 7-4 (282.99 mg, 917.68 umol), N,N-carbonyldiimidazole (148.80 mg, 917.68 umol), compound 1-5 (130.00 mg, 229.42 umol), N-methylmorpholine (92.82 mg, 917.68 umol), 4-dimethylaminopyridine (5.61 mg, 45.88 umol), and DMF (3 mL) were added. The mixture was heated to 80°C and stirred for 16 hours until the starting material disappeared by LC-MS. The reaction system was then directly analyzed by MPLC to yield compound 7-5 (136.00 mg, 65.79% yield). LCMS (E+) m / z: 901.3 [M+H] + .

[0565] Step 5. Synthesis of compound 7-6

[0566] To a 25 mL single-necked vial, add compound 7-5 (136.00 mg, 150.94 μmol), followed by dioxane hydrochloride (6 mL). Stir the mixture at room temperature for 2.5 hours, monitored by LC-MS. After completion of the reaction, concentrate the mixture to yield compound 7-6 (130.00 mg, crude product).

[0567] Step 6. Synthesis of compound 7

[0568] Compound 7-6 (130.00 mg, 162.32 μmol) was added to a 25 mL reaction flask and dissolved in 5 mL of methanol:water (3:1). Lithium hydroxide monohydrate (40.90 mg, 973.91 μmol) was then added. The reaction was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, the reaction mixture was directly analyzed by MPLC to yield compound 7 (62.00 mg, 48.54% yield).

[0569] LCMS (E+) m / z: 787.2 [M+H] + .

[0570] 1H NMR (600MHz, Methanol-d4) δ8.34(d,J=8.4Hz,1H),7.75(d,J=8.5Hz,1H),7.69(d,J=6.1Hz,1H),7.51–7.43(m,3H),7 .41–7.36(m,1H),7.32(d,J=8.0Hz,2H),7.22(d,J=7.8Hz,1H),6.96(d,J=7.9Hz,1H),6.55(d,J=8.7Hz,1H),6.47(d,J =7.7Hz,1H),5.35–5.15(m,1H),4.42–4.24(m,2H),4.10–3.96(m,2H),3.83–3.76(m,2H),3.73–3.69(m,2H),3.69–3.6 3(m,2H),3.63–3.54(m,8H),3.43–3.31(m,4H),2.83–2.81(m,2H),2.49–2.37(m,2H),2.22(s,3H),2.07–1.94(m,2H).

[0571] Example 8

[0572] Synthesis of compound 8

[0573] Step 1. Synthesis of compound 8-2

[0574] Under nitrogen protection, compound 8-1 (2.00 g, 12.98 mmol), 8-1' (2.98 g, 14.29 mmol), potassium carbonate (3.60 g, 25.90 mmol), and DMF (20 mL) were added. After addition, the mixture was stirred at room temperature for 3 hours and monitored by LC-MS. After completion of the reaction, water (500 mL) was added, and the mixture was extracted with ethyl acetate, washed with saturated brine, and dried over anhydrous sodium sulfate. Compound 8-2 (2.00 g, 54.55% yield) was obtained by reverse phase preparation. LCMS (E+) m / z: 283.1 [M+H] + .

[0575] Step 2. Synthesis of compound 8-3

[0576] Under nitrogen protection, compound 8-2 (2.00 g, 7.09 mmol) was added to a 100 mL reaction flask and dissolved in ethanol (20 mL). Acetic acid (5 mL) and iron powder (2.80 g, 50.00 mmol) were then added. The reaction was allowed to proceed at room temperature for 5 hours and monitored by LC-MS. After the reaction, the system was filtered and concentrated using celite, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and then analyzed by MPLC to obtain compound 8-3 (1.78 g, 99.62% yield). LCMS (E+) m / z: 253.0 [M+H] + .

[0577] Step 3. Synthesis of compound 8-4

[0578] Under nitrogen protection, compound 8-3 (1.78 g, 7.06 mmol) was added to a 100 mL three-necked flask and dissolved with DCE (50 mL). Then, sodium triacetoxyborohydride (1.65 g, 7.81 mmol) and acetic acid (5 drops) were added. After the addition, the reaction was allowed to react at room temperature overnight and monitored by LC-MS. After the reaction was completed, the pH was adjusted to 5 with hydrochloric acid, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. Compound 8-4 (1.00 g, 60.00% yield) was obtained by MPLC. LCMS (E+) m / z: 238.2 [M+H] + .

[0579] Step 4. Synthesis of compound 8-5

[0580] Under nitrogen, compound 8-4 (1.00 g, 4.24 mmol) was added to a 50 mL reaction flask and dissolved in methanol (10 mL). Sodium hydroxide solution (1 M, 10 mL) was then added. The reaction was allowed to react at room temperature for 2 hours, monitored by LC-MS. After the reaction, the methanol was concentrated to remove the methanol, and the residue was adjusted to pH 1 with hydrochloric acid. MPLC analysis yielded the hydrochloride salt of compound 8-5 (1.00 g, 91.41% yield). LCMS (E+) m / z: 223.3 [M+H] + .

[0581] Step 5. Synthesis of compound 8-6

[0582] Under nitrogen, compound 8-5 (50.00 mg, 0.19 mmol) and 1-7 (114.00 mg, 0.19 mmol) were added to a 4 mL reaction vial and dissolved in N,N'-dimethylformamide (2 mL). 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (73.00 mg, 0.19 mmol) and N,N-diisopropylethylamine (68.00 mg, 0.60 mmol) were added to the system with stirring. After addition, the reaction was allowed to react at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, the reaction solution was purified by MPLC to obtain compound 8-6 (80 mg, 50.91% yield). LCMS (E+) m / z: 828.7 [M+H] + .

[0583] Step 6. Synthesis of compound 8

[0584] Compound 8-6 (80.00 mg, 0.10 mmol) was added to a 20 mL reaction flask and dissolved in methanol (2 mL). 1 M sodium hydroxide solution (2 mL) was then added. The reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, compound 8 (50.00 mg, 61.50% yield) was obtained directly by MPLC. LCMS (E+) m / z: 814. [M+H] + .

[0585] Synthesis of compounds 8A and 8B

[0586] Compound 8-5 was chiral resolved by SFC to give compounds 8-5A and 8-5B. Compounds 8A and 8B were prepared using the same synthetic route as above.

[0587] 8A

[0588] LCMS (E+) m / z: 814 [M+H] + .

[0589] 11H NMR (600 MHz, DMSO-d6) δ 8.98 (s, 1H), 8.30–8.20 (m, 2H), 7.77 (d, J = 8.3 Hz, 1H), 7.51 (dt, J = 15.1, 8.1 Hz, 2H), 7.44 (d, J = 7.8 Hz, 2H), 7.41 (d, J = 4.9 Hz, 1H), 7.31 (dd, J = 23.7, 7.8 Hz, 3H), 7.03 (d, J = 8.0 Hz, 1H), 6.70 (s, 1H), 6.33 (d, J = 5.0 Hz, 1H), 5.23 (s, 1H), 4.31 (d, J = 5.2 Hz, 2H), 4.10 (dd, J = 10.6, 2.8 Hz, 1H), 3.92 (d, J = 4.7 Hz, 2H), 3.81 (dd, J = 10.7, 5.9 Hz, 1H), 3.74 (d, J = 5.8 Hz, 2H), 3.68 (dd, J = 5.8, 3.8 Hz, 2H), 3.63–3.48 (m, 10H), 3.42 (s, 1H), 3.38–3.35 (m, 2H), 2.60 (d, J = 14.0 Hz, 3H), 2.32 (t, J = 7.6 Hz, 2H), 2.03 (s, 3H), 1.72 (m, 2H). 8B

[0590] LCMS (E+) m / z: 814 [M+H] + .

[0591] 1 1H NMR (600 MHz, DMSO-d6) δ 8.85 (s, 1H), 8.25 (t, J = 15.4 Hz, 2H), 7.77 (d, J = 8.1 Hz, 1H), 7.69–7.24 (m, 8H), 7.04 (d, J = 8.3 Hz, 1H), 6.71 (s, 1H), 6.33 (t, J = 4.2 Hz, 1H), 5.24 (s, 1H), 4.34–4.27 (m, 2H), 4.10 (d, J = 10.6 Hz, 1H), 3.92 (t, J = 4.8 Hz, 2H), 3.85–3.63 (m, 5H), 3.62–3.49 (m, 十三H), 3.42 (s, 1H), 3.36 (t, J = 4.9 Hz, 2H), 2.62 (d, J = 11.9 Hz, 2H), 2.39–2.27 (m, 2H), 2.02 (d, J = 4.2 Hz, 3H), 1.83–1.64 (m, 2H).

[0592] Example 9

[0593] Synthesis of Compound 9

[0594] Step 1. Synthesis of compound 9-2

[0595] Compound 9-1 (250 mg, 1.22 mmol) was added to a 4 mL reaction vial and dissolved in methanol (2 mL). H2SO4 (36.02 mg, 367.24 μmol) was added dropwise to the reaction system. After addition, the temperature was raised to 60°C and stirred for 4 hours, monitored by LC-MS. After completion of the reaction, the reaction solution was directly analyzed by MPLC to yield compound 9-2 (240 mg, 89.83% yield). LCMS (E+) m / z: 219.1 [M+H] + .

[0596] Step 2. Synthesis of compound 9-3

[0597] Compound 9-2 (240 mg, 1.10 mmol) was added to a 10 mL reaction flask and dissolved in DMF (3 mL). The temperature was lowered to 0-5°C and NaH (131.96 mg, 3.30 mmol, 60% mass fraction) was added portionwise. After addition, the reaction was stirred at this temperature for 30 minutes. (Boc)2O (360.00 mg, 1.65 mmol) was then added dropwise to the reaction system. After addition, the temperature was raised to 20°C and the reaction was stirred for 1 hour, monitored by LC-MS. After completion of the reaction, the reaction solution was added dropwise to saturated ammonium chloride (50 mL), extracted twice with ethyl acetate (60 mL), washed with saturated brine, dried over anhydrous sodium sulfate, and filtered to dryness to obtain compound 9-3 (350 mg, 99.97% yield). LCMS (E+) m / z: 319.3 [M+H] + .

[0598] Step 3. Synthesis of compound 9-4

[0599] Compound 9-3 (350 mg, 1.10 mmol) was added to a 100 mL reaction flask and dissolved in methanol (5 mL). Palladium on carbon (201.61 mg, 166.00 μmol) was then added. The reaction system was purged with hydrogen three times and stirred at room temperature for 16 hours, monitored by LC-MS. After completion of the reaction, the mixture was filtered and concentrated to afford compound 9-4 (250 mg, 70.98% yield). LCMS (E+) m / z: 321.1 [M+H] + .

[0600] Step 4. Synthesis of compound 9-5

[0601] Compound 9-4 (250 mg, 780.33 μmol) was added to a 4 mL reaction vial and dissolved in methanol (1 mL). 1 M sodium hydroxide solution (1 mL) was then added. The reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, compound 9-5 (238 mg, 99.35% yield) was obtained directly by MPLC. LCMS (E+) m / z: 307.1 [M+H] + .

[0602] Step 5. Synthesis of compound 9-6

[0603] To a 4 mL reaction vial, compound 9-5 (238 mg, 734.45 μmol) and compound 1-7 (458.08 mg, 734.45 μmol) were added and dissolved in dry DMF (5 mL). HATU (362.83 mg, 954.78 μmol) and DIPEA (284.23 mg, 2203.33 μmol) were added with stirring. After addition, the reaction was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, compound 9-6 (425 mg, 63.45% yield) was obtained directly by MPLC. LCMS (E+) m / z: 912.2 [M+H] + .

[0604] Step 6. Synthesis of compound 9-7

[0605] Compound 9-6 (220 mg, 240.12 μmol) was added to a 4 mL reaction vial and dissolved in 4 M dioxane hydrochloride (0.5 mL). The reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, compound 9-7 (170 mg, 86.80% yield) was obtained directly by MPLC. LCMS (E+) m / z: 812.8 [M+H] + .

[0606] Step 7. Synthesis of compound 9

[0607] Compound 9-7 (170 mg, 200.96 μmol) was added to a 4 mL reaction vial and dissolved in methanol (1 mL). 1 M sodium hydroxide solution (1 mL) was then added. The reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, compound 9 (150 mg, 83.60% yield) was obtained directly by MPLC.

[0608] LCMS (E+) m / z: 798.9 [M+H] + .

[0609] Synthesis of compounds 9A and 9B

[0610] Compound 9-5 was chiral resolved by SFC to give compounds 9-5A and 9-5B. Compounds 9A and 9B were prepared using the same synthetic route as above.

[0611] 9A

[0612] LCMS (E+) m / z: 798.9 [M+H] + .

[0613] 1H NMR (400MHz, DMSO-d6) δ8.51(d,J=8.3Hz,1H),8.27(dd,J=7.5,2.1Hz,1H),8.15(t,J=5.8Hz,1H),7.87–7.71(m,2H),7.63–7.3 5(m,7H),7.32(d,J=7.9Hz,1H),7.05(d,J=8.0Hz,1H),6.41(d,J=6.1Hz,1H),5.31(q,J=7.5Hz,1H),4.33(dd,J=5.8,3.4Hz,2H ),3.93(dd,J=5.7,3.4Hz,2H),3.86–3.65(m,4H),3.63–3.51(m,12H),3.36(t,J=4.9Hz,2H),3.20–3.06(m,1H),2.93(q,J=7.3 Hz,1H),2.79(d,J=7.3Hz,2H),2.63(dd,J=17.0,6.5Hz,1H),2.26(t,J=7.5Hz,2H),2.11(s,3H),1.80(dh,J=28.4,6.9Hz,2H).

[0614] 9B

[0615] LCMS (E+) m / z: 798.9 [M+H] + .

[0616] 1H NMR(400MHz,Chloroform-d)δ8.92(s,1H),8.40–8.28(m,1H),7.83(d,J=8.4Hz,1H),7.50–7.31(m,6H),7.22(d,J= 7.9Hz,1H),7.15(d,J=6.5Hz,1H),7.07–6.92(m,1H),6.83(d,J=8.0Hz,1H),6.19(d,J=6.5Hz,1H),5.41(q,J=6.1Hz ,1H),4.30(dt,J=21.9,4.3Hz,3H),4.14–3.89(m,5H),3.82(dd,J=5.9,3.6Hz,2H),3.74–3.60(m,13H),3.36(t,J= 5.1Hz,2H),3.14(dd,J=17.1,9.3Hz,1H),2.82(qd,J=14.7,5.4Hz,2H),2.69–2.43(m,3H),2.06(d,J=39.4Hz,5H)..

[0617] Example 10

[0618] Synthesis of compound 10

[0619] Step 1. Synthesis of compound 10-2

[0620] Under ice, 1,2-dichloroethane (97 mL) was added to a 250 mL single-necked flask, followed by 2-amino-4-methylpyridine (4.50 g, 18.20 mmol) and stirred to fully dissolve. Sodium acetate borohydride (7.71 g, 36.39 mmol) and compound 10-1' (2.36 g, 21.84 mmol) were slowly added to the above system in sequence. The reaction system was returned to room temperature under nitrogen and stirred for 16 hours. The reaction system was diluted with saturated sodium carbonate solution, extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, and the organic layer was concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:2) to obtain compound 10-2 (5.84 g, 94.55% yield). LCMS (E+) m / z: 340.2 [M+H] + .

[0621] Step 2. Synthesis of compound 10-3

[0622] At room temperature, dichloromethane (80 mL) was added to a 250 mL single-necked flask, followed by compound 10-2 (6.00 g, 17.68 mmol) and stirred to fully dissolve. Triethylamine (1.79 g, 17.68 mmol) and di-tert-butyl dicarbonate (7.72 g, 35.35 mmol) were slowly added to the above system in sequence, and the reaction system was stirred at room temperature for 16 hours. The reaction system was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound 10-3 (6.14 g, 79.02% yield). LCMS (E+) m / z: 440.2 [M+H] + .

[0623] Step 3. Synthesis of compound 10-4

[0624] To a 250 mL single-necked flask, add intermediate 10-3 (6.14 g, 13.97 mmol), followed by anhydrous methanol (100 mL) and 10% palladium on carbon (1.23 g). After the additions were complete, the reaction system was stirred at room temperature under a hydrogen balloon for 3 hours, monitored by LC-MS. After completion of the reaction, the mixture was concentrated to afford intermediate 10-4 (4.21 g, crude product). LCMS (E+) m / z: 306.4 [M+H] + .

[0625] Step 4. Synthesis of compound 10-5

[0626] Compound 10-4 (2.14 g, 7.01 mmol) was added to a 100 mL reaction flask and dissolved in DMF (21 mL). Methyl bromoacetate (1.18 g, 7.71 mmol) and potassium carbonate (2.42 g, 17.52 mmol) were then added. The reaction was stirred at room temperature for 16 hours and monitored by LC-MS. After completion of the reaction, the reaction system was diluted with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and the organic layer was concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain compound 10-5 (1.98 g, 74.86% yield). LCMS (E+) m / z: 378.6 [M+H] + .

[0627] Step 5. Synthesis of compound 10-6

[0628] Compound 10-5 (1.86 g, 4.93 mmol) was added to a 25 mL reaction flask and dissolved in anhydrous methanol (9 mL) and water (3 mL). Lithium hydroxide monohydrate (1.03 g, 24.64 mmol) was then added. The reaction mixture was stirred at room temperature for 2 hours and monitored by LC-MS. After the reaction was complete, the reaction solution was directly analyzed by MPLC to obtain compound 10-6 (1.52 g, 84.87% yield). LCMS (E+) m / z: 364.2 [M+H] + .

[0629] Step 6. Synthesis of compound 10-7

[0630] To a 20 mL single-necked flask, intermediate 10-6 (212.00 mg, 583.30 umol), N,N-diisopropylethylamine (264.57 mg, 2.05 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (291.72 mg, 767.68 umol), and DMF (4 mL) were added. The system was stirred at room temperature for 10 minutes, and then compound 1-5 (290.00 mg, 511.79 umol) was added. After the addition, the mixture was allowed to react at room temperature for 2 hours until the starting material disappeared as determined by LC-MS. The reaction solution was directly subjected to MPLC to obtain compound 10-7 (448.00 mg, 95.97% yield). LCMS (E+) m / z: 912.8 [M+H] + .

[0631] Step 7. Synthesis of compound 10-8

[0632] To a 25 mL single-necked flask, add intermediate 10-7 (448.00 mg, 491.19 μmol), followed by dichloromethane (2 mL) and 4 M hydrochloric acid in dioxane (8 mL). After complete addition, stir at room temperature for 2.5 hours and monitor with LC-MS. After completion of the reaction, concentrate to yield compound 10-8 (399.00 mg, crude product).

[0633] Step 8. Synthesis of compound 10

[0634] Compound 10-8 (399 mg, 491.4 μmol) was added to a 25 mL reaction flask and dissolved in anhydrous methanol (3 mL) and water (1 mL). Lithium hydroxide monohydrate (123.83 mg, 2.95 mmol) was then added. The reaction mixture was stirred at room temperature for 2 hours and monitored by LC-MS. After the reaction was complete, the reaction solution was directly analyzed by MPLC to obtain compound 10 (190 mg, 48.46% yield). LCMS (E+) m / z: 798.2 [M+H] + . 1H NMR (600MHz, Methanol-d4) δ8.31(d,J=8.5Hz,1H),7.73(d,J=8.5Hz,1H),7.69(d,J=5.7Hz,1H),7.46(d,J=7.9Hz,2H),7.44–7.40(m,1H),7.34( t,J=8.8Hz,3H),7.19(d,J=7.8Hz,1H),6.88(d,J=7.9Hz,1H),6.48(s,1H ),6.43(d,J=5.2Hz,1H),5.41(t,J=6.3Hz,1H),4.31–4.26(m,2H),3.99– 3.94(m,2H),3.78–3.73(m,2H),3.70–3.65(m,2H),3.64–3.60(m,2H),3 .60–3.51(m,8H),3.27(t,J=4.9Hz,2H),3.21(s,2H),3.16(dd,J=6.7,4. 2Hz,2H),3.03(d,J=15.2Hz,2H),2.81(d,J=7.4Hz,2H),2.40–2.27(m,2H ),2.21(s,3H),1.86–1.73(m,2H),1.69–1.62(m,1H),1.52–1.45(m,2H).

[0635] Example 11

[0636] Synthesis of compound 11

[0637] Step 1. Synthesis of compound 11-2

[0638] At room temperature, tert-butyl alcohol (72 mL) was added to a 250 mL single-necked flask, followed by compound 11-1 (4.53 g, 24.22 mmol) and stirred to fully dissolve. Triethylamine (7.35 g, 72.66 mmol), 4-dimethylaminopyridine (591.79 mg, 4.84 mmol), and di-tert-butyl dicarbonate (15.86 g, 72.66 mmol) were slowly added to the above system, and the reaction system was stirred at 40 degrees for 16 hours. The reaction system was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1) to obtain compound 11-2 (6.54 g, 69.73% yield). LCMS (E+) m / z: 387.1 [M+H] + .

[0639] Step 2. Synthesis of compound 11-3

[0640] At room temperature, DMF (26 mL) was added to a 100 mL single-necked flask, followed by compound 11-2 (3.47 g, 8.96 mmol) and stirred to fully dissolve. Methyl acrylate (3.09 g, 35.84 mmol), palladium acetate (201.17 mg, 896.02 mmol), N,N-diisopropylethylamine (11.58 mg, 89.60 mmol), and tri(o-tolyl)phosphine (409.08 mg, 1.34 mmol) were slowly added to the above system. The reaction system was stirred at 100°C under nitrogen for 16 hours. The reaction system was diluted with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and the organic layer was concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 4:1) to obtain compound 11-3 (3.21 g, 91.29% yield). LCMS (E+) m / z: 393.2 [M+H] + .

[0641] Step 3. Synthesis of compound 11-4

[0642] To a 100 mL single-necked flask, intermediate 11-3 (870 mg, 2.22 mmol) was added, followed by anhydrous methanol (30 mL) and 10% palladium on carbon (100 mg). After the additions were complete, the reaction system was stirred at room temperature under a hydrogen balloon for 16 hours, monitored by LC-MS. After completion of the reaction, the mixture was concentrated to afford intermediate 11-4 (810 mg, crude product). LCMS (E+) m / z: 395.5 [M+H] + .

[0643] Step 4. Synthesis of compound 11-5

[0644] Compound 11-4 (738 mg, 1.87 mmol) was added to a 20 mL reaction flask and dissolved in anhydrous methanol (3 mL) and water (1 mL). Lithium hydroxide monohydrate (78.58 mg, 1.87 mmol) was then added. The reaction was stirred at room temperature for 2 hours and monitored by LC-MS. After the reaction was complete, the reaction solution was directly analyzed by MPLC to obtain compound 11-5 (300.00 mg, 57.20% yield). LCMS (E+) m / z: 281.1 [M+H] + .

[0645] Step 5. Synthesis of compound 11-6

[0646] To a 20 mL single-necked flask, intermediate 11-5 (210.00 mg, 749.15 umol), N,N-diisopropylethylamine (387.28 mg, 3.00 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (427.01 mg, 1.12 mmol), and DMF (5.50 mL) were added. The system was stirred at room temperature for 10 minutes, and then compound 1-7 (327.07 mg, 524.40 umol) was added. After the addition, the mixture was allowed to react at room temperature for 2 hours until the starting material disappeared as determined by LC-MS. The reaction solution was directly subjected to MPLC to obtain compound 11-6 (467.00 mg, 70.36% yield). LCMS (E+) m / z: 886.8 [M+H] + .

[0647] Step 6. Synthesis of compound 11-7

[0648] To a 50 mL single-necked flask, add intermediate 11-6 (467.00 mg, 527.09 μmol), followed by dichloromethane (2 mL) and 4 M hydrochloric acid in dioxane (8 mL). After complete addition, stir at room temperature for 2.5 hours and monitor with LC-MS. After completion of the reaction, concentrate to yield compound 11-7 (378.00 mg, crude product).

[0649] Step 7. Synthesis of compound 11

[0650] Compound 11-7 (378 mg, 480.99 umol) was added to a 20 mL reaction flask and dissolved in anhydrous methanol (3 mL) and water (1 mL). Lithium hydroxide monohydrate (121.21 mg, 2.89 mmol) was then added. The reaction was stirred at room temperature for 2 hours and monitored by LC-MS. After the reaction was complete, the reaction solution was directly analyzed by MPLC to obtain compound 11 (218.00 mg, 58.72% yield). LCMS (E+) m / z: 772.1 [M+H] + . 1H NMR(600MHz, Methanol-d4)δ8.32(d,J=7.9Hz,1H),7.77(d,J=8.3Hz,1H),7.49–7.41(m,4H),7.40–7.33(m,3H), 7.23(d,J=7.9Hz,1H),6.92(d,J=8.0Hz,1H),6.54(d,J=7.3Hz,1H),5.24(q,J=5.5Hz,1H),4.31(t,J=4.7Hz,2H), 4.07–3.95(m,3H),3.84–3.75(m,3H),3.67(dd,J=5.9,3.4Hz,2H),3.62(dd,J=6.1,3.3Hz,2H),3.59–3.53(m,8H) ,3.28(t,J=5.0Hz,2H),3.05–3.00(m,2H),2.86–2.82(m,1H),2.78–2.71(m,2H),2.70–2.60(m,1H),2.08(s,3H).

[0651] Example 12

[0652] Synthesis of compound 12

[0653] Step 1. Synthesis of compound 12-2

[0654] Under ice bath, DMF (15 mL) and Boc-2-amino-3-propionic acid methyl ester pyridine (1.37 g, 4.89 mmol) were added to a 100 mL three-necked flask and stirred to fully dissolve. Sodium hydride (235.00 mg, 5.87 mmol) was slowly added to the system and the reaction was continued for 30 minutes. Then iodomethane (903.00 mg, 6.36 mmol) was added and the system was moved to room temperature for 2 hours. LC-MS monitored the complete reaction of the raw material. Extracted with water and ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated, and prepared by MPLC to obtain compound 12-2 (900.00 mg, 63.77% yield). LCMS (E+) m / z: 295.2 [M+H] + .

[0655] Step 2. Synthesis of compound 12-3

[0656] Compound 12-2 (900.00 mg, 3.06 mmol) was dissolved in 10 mL of methanol, and sodium hydroxide solution (2N, 10 mL) was added dropwise. The reaction system was stirred at 25°C for 1 hour. After the reaction, the methanol in the system was concentrated, and the pH was adjusted to 5.0 with 3M hydrochloric acid. The aqueous phase was purified by MPLC to obtain compound 12-3 (800.00 mg, 93.37% yield). LCMS (E+) m / z: 281.1 [MH] + .

[0657] Step 3. Synthesis of compound 12-4

[0658] Compound 12-3 (56.00 mg, 0.20 mmol), intermediate 1-7 (93.00 mg, 0.15 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (95.00 mg, 0.23 mmol), and N,N-diisopropylethylamine (100 μL, 0.60 mmol) were added to a 4 mL reaction vial. After addition, the mixture was allowed to react at room temperature for 2 hours until the starting material disappeared completely as determined by LC-MS. The reaction solution was directly analyzed by MPLC to yield compound 12-4 (85.00 mg, 48.01% yield). LCMS (E+) m / z: 886.8 [M+H] + .

[0659] Step 4. Synthesis of compound 12-5

[0660] Compound 12-4 (85.00 mg, 0.096 mmol) was added to a 25 mL reaction flask, followed by dissolution with methanol (5 mL). Sodium hydroxide solution (2N, 5 mL) was then added dropwise. The reaction system was stirred at 25°C for 1 hour. After the reaction, the methanol in the system was concentrated, and the pH was adjusted to 5.0 with 3 M hydrochloric acid. The product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to afford compound 12-5 (85.00 mg, 100% yield). LCMS (E+) m / z: 872.4 [M+H] + .

[0661] Step 5. Synthesis of compound 12

[0662] Compound 12-5 (85.00 mg, 0.096 mmol) was added to a 20 mL single-necked vial and dissolved in dioxane hydrochloride (4.0 M, 5 mL). The mixture was allowed to react at room temperature for 2 hours until the starting material disappeared as determined by LC-MS. The reaction mixture was directly analyzed by MPLC to yield compound 12 (60.00 mg, 77.52% yield).

[0663] LCMS (E+) m / z: 772.2 [M+H] + .

[0664] 1 H NMR (600MHz, DMSO-d6) δ8.61(s,1H),8.30–8.24(m,1H),8.19(t,J=5.8Hz,1H),7.82–7.72(m,1H),7.58–7.47(m,2H),7.45(d,J=7.8Hz ,2H),7.38(d,J=7.3Hz,2H),7.32(t,J=6.0Hz,1H),7.26(t,J=7.7Hz,1H),7.05(d,J=7.9Hz,1H),6.34(d,J=7.2Hz,2H),6.22(d,J=8.3H z,1H),5.28(q,J=7.5Hz,1H),4.32(t,J=4.6Hz,2H),3.92(t,J=4.6Hz,2H),3.74(dd,J=5.8,2.7Hz,2H),3.69(dd,J=5.8,3.8Hz,2H),3 .58(dt,J=11.9,5.1Hz,4H),3.54(d,J=5.1Hz,2H),3.52(t,J=2.2Hz,6H),3.36(t,J=4.9Hz,3H),2.78–2.68(m,7H),2.51–2.53(m,3H).

[0665] Example 13

[0666] Synthesis of compound 13

[0667] Step 1. Synthesis of compound 13-2

[0668] At room temperature, DCE (50 mL) was added to a 250 mL single-necked flask, followed by compound 13-1 (1.09 g, 5.16 mmol), and stirred to fully dissolve. 2-Amino-4-methylpyridine (836.94 mg, 7.74 mmol) and tetraisopropyl titanate (2.20 g, 7.74 mmol) were slowly added to the above system. The system was stirred at room temperature for 30 minutes, followed by sodium acetate borohydride (5.47 g, 25.80 mmol). After the addition, the reaction system was stirred at 70°C for 2 hours. The reaction system was diluted with saturated sodium bicarbonate, extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, and the organic layer was concentrated. The crude product was purified by MPLC to yield compound 13-2 (696 mg, 44.46% yield). LCMS (E+) m / z: 304.2 [M+H] + .

[0669] Step 2. Synthesis of compound 13-3

[0670] To a 50 mL single-necked flask, add intermediate 13-2 (696.00 mg, 2.29 mmol), followed by dichloromethane (2 mL) and a 4 M hydrochloric acid solution in dioxane (8 mL). After complete addition, stir at room temperature for 2.5 hours and monitor with LC-MS. After completion of the reaction, concentrate to afford compound 13-3 (466 mg, crude product).

[0671] Step 3. Synthesis of compound 13-4

[0672] Intermediate 13-3 (466 mg, 2.29 mmol) was added to a 25 mL single-necked flask and dissolved in DMF (7 mL). DBU (1.05 g, 6.88 mmol) and methyl bromoacetate (420.81 mg, 2.75 mmol) were then added sequentially. The reaction system was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, the reaction solution was directly analyzed by MPLC to yield compound 13-4 (500.00 mg, 79.21% yield). LCMS (E+) m / z: 276.2 [M+H] + .

[0673] Step 4. Synthesis of compound 13-5

[0674] Compound 13-4 (500 mg, 1.82 mmol) was added to a 25 mL reaction flask and dissolved in anhydrous methanol (4.50 mL) and water (1.50 mL). Lithium hydroxide monohydrate (457.61 mg, 10.90 mmol) was then added. The reaction mixture was stirred at room temperature for 2 hours and monitored by LC-MS. After the reaction was completed, the reaction solution was directly analyzed by MPLC to obtain compound 13-5 (455.00 mg, 95.88% yield). LCMS (E+) m / z: 262.2 [M+H] + .

[0675] Step 5. Synthesis of compound 13-6

[0676] To a 10 mL single-necked vial, intermediate 15-5 (60.00 mg, 229.60 umol), N,N-diisopropylethylamine (118.70 mg, 918.42 umol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (130.87 mg, 344.41 umol), and DMF (3 mL) were added. The system was stirred at room temperature for 10 minutes, and then compound 1-5 (130.10 mg, 229.60 umol) was added. After the addition, the mixture was allowed to react at room temperature for 2 hours until the starting material disappeared as determined by LC-MS. The reaction solution was directly subjected to MPLC to afford compound 13-6 (130.00 mg, 69.90% yield). LCMS (E+) m / z: 810.8 [M+H]+ .

[0677] Step 6. Synthesis of compound 13

[0678] Compound 13-6 (130.00 mg, 160.50 μmol) was added to a 20 mL reaction flask and dissolved in anhydrous methanol (3 mL) and water (1 mL). Lithium hydroxide monohydrate (40.45 mg, 963.03 μmol) was then added. The reaction mixture was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, the reaction mixture was directly analyzed by MPLC to yield compound 13 (82.00 mg, 64.19% yield).

[0679] LCMS (E+) m / z: 796.8 [M+H] + .

[0680] 1 H NMR(600MHz, Methanol-d4)δ8.35(d,J=8.5Hz,1H),7.74(dd,J=14.3,7.7Hz,2H),7.50–7.45(m,3H),7.44–7.40(m,3H ),7.28(d,J=7.8Hz,1H),6.98(d,J=7.9Hz,1H),6.80–6.77(m,2H),5.45(dd,J=8.2,6.5Hz,1H),4.36(dd,J=5.6,3.6Hz ,4H),4.25(s,2H),4.11(t,J=6.9Hz,2H),4.02(dd,J=5.5,3.6Hz,2H),3.80(dd,J=5.8,3.5Hz,2H),3.70(dd,J=5.7,3. 5Hz,2H),3.65(dd,J=6.1,3.4Hz,2H),3.62–3.56(m,9H),3.30(d,J=4.9Hz,2H),2.97–2.88(m,4H),2.41–2.34(m,5H).

[0681] Example 14

[0682] Synthesis of compound 14

[0683] Step 1. Synthesis of compound 14-2

[0684] Compound 14-1 (7.26 g, 41.97 mmol) was added to a 250 mL reaction flask and dissolved in acetonitrile (100 mL). 1-(Trifluoromethyl)-1,2-benzidoxyl-3(1H)-one (4.42 g, 13.99 mmol) and K2CO3 (2.90 g, 20.98 mmol) were added to the reaction system. After the addition, the temperature was raised to 75°C and stirred for 6 hours, monitored by LC-MS. After the reaction was completed, the reaction solution was dried and the crude product was purified by MPLC to obtain compound 14-2 (1.05 g, 31.14% yield). LCMS (E+) m / z: 241.2 [M+H] + .

[0685] Step 2. Synthesis of compound 14-3

[0686] To a 20 mL reaction flask, compound 14-2 (480 mg, 1.99 mmol), methyl acrylate (685.83 mg, 7.97 mmol), Pd(OAc)2 (89.43 mg, 398.33 μmol), P(o-tolyl)3 (90.93 mg, 298.74 μmol), and DIEA (2.57 g, 19.92 mmol) were added and dissolved in DMF (5 mL). The reaction system was purged with nitrogen three times, heated to 100°C, and stirred for 16 hours, monitored by LC-MS. After the reaction, the filtered filtrate was purified by MPLC to obtain compound 14-3 (370 mg, 75.46% yield). LCMS (E+) m / z: 247.2 [M+H] + .

[0687] Step 3. Synthesis of compound 14-4

[0688] Compound 14-3 (370 mg, 1.50 mmol) was added to a 25 mL reaction flask and dissolved in methanol (5 mL). Palladium on carbon (275.62 mg, 226.94 μmol) was then added. The reaction system was purged with hydrogen three times and stirred at room temperature for 16 hours, monitored by LC-MS. After completion of the reaction, the mixture was filtered and concentrated to afford compound 14-4 (260 mg, 69.70% yield). LCMS (E+) m / z: 249.1 [M+H] + .

[0689] Step 4. Synthesis of compound 14-5

[0690] Compound 14-4 (160 mg, 644.64 μmol) was added to a 4 mL reaction vial and dissolved in methanol (2 mL). 1 M sodium hydroxide solution (1 mL) was then added. The reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After the reaction was complete, compound 14-5 (240 mg, crude) was obtained directly by MPLC. LCMS (E+) m / z: 235.0 [M+H] + .

[0691] Step 5. Synthesis of compound 14-6

[0692] To a 4 mL reaction vial, compound 14-5 (30 mg, 128.11 μmol) and compound 1-7 (79.90 mg, 128.11 μmol) were added and dissolved in dry DMF (2 mL). HATU (56.29 mg, 166.54 μmol) and DIPEA (49.58 mg, 384.33 μmol) were added with stirring. After addition, the reaction was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, compound 14-6 (50 mg, 46.47% yield) was obtained directly by MPLC. LCMS (E+) m / z: 840.8 [M+H] + .

[0693] Step 6. Synthesis of compound 14

[0694] Compound 14-6 (40 mg, 47.63 μmol) was added to a 4 mL reaction vial and dissolved in methanol (1 mL). 1 M sodium hydroxide solution (1 mL) was then added. The reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, compound 14 (20 mg, 50.42% yield) was obtained directly by MPLC. LCMS (E+) m / z: 826.3 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ12.35(s,1H),8.47(d,J=8.3Hz,1H),8.30–8.24(m,1H),8.18(t,J=5.8Hz,1H),7.79–7.75(m,1H ),7.65(d,J=7.9Hz,1H),7.52(ddd,J=7.7,5.5,1.7Hz,2H),7.45(d,J=8.3Hz,2H),7.40(d,J=8.2Hz,2H),7.33(d,J=7.9H z,1H),7.06(d,J=8.1Hz,1H),6.55(d,J=7.8Hz,1H),6.38(s,2H),5.30(q,J=7.5Hz,1H),4.33(dd,J=5.7,3.6Hz,2H),3. 97–3.88(m,2H),3.75(d,J=5.8Hz,2H),3.69(dd,J=5.9,3.6Hz,2H),3.64–3.10(m,13H),2.86–2.76(m,4H),2.51(m,3H).

[0695] Example 15

[0696] Synthesis of compound 15

[0697] Step 1. Synthesis of compound 15-2

[0698] To a 20 mL reaction flask, compound 15-1 (380 mg, 1.99 mmol), methyl acrylate (685.10 mg, 7.96 mmol), Pd(OAc)2 (89.33 mg, 397.90 μmol), P(o-tolyl)3 (90.83 mg, 298.43 μmol), and DIEA (2.57 g, 19.92 mmol) were added and dissolved in DMF (5 mL). The reaction system was purged with nitrogen three times, heated to 100°C, and stirred for 16 hours, monitored by LC-MS. After completion of the reaction, the filtrate was filtered and analyzed by MPLC to yield compound 15-2 (380 mg, 97.36% yield). LCMS (E+) m / z: 197.0 [M+H] + .

[0699] Step 2. Synthesis of compound 15-3

[0700] Compound 15-2 (350 mg, 1.78 mmol) was added to a 25 mL reaction flask and dissolved in methanol (5 mL). Palladium on carbon (327.18 mg, 269.40 μmol) was then added. The reaction system was purged with hydrogen three times and stirred at room temperature for 16 hours, monitored by LC-MS. After completion of the reaction, the mixture was filtered and concentrated to afford compound 15-3 (300 mg, 84.84% yield). LCMS (E+) m / z: 199.0 [M+H] + .

[0701] Step 3. Synthesis of compound 15-4

[0702] Compound 15-3 (200 mg, 1.01 mmol) was added to a 4 mL reaction flask and dissolved in methanol (2 mL). 1 M sodium hydroxide solution (1 mL) was then added. After addition, the reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, compound 15-4 (300 mg, crude product) was obtained directly by MPLC. LCMS (E+) m / z: 185.1 [M+H] + .

[0703] Step 4. Synthesis of compound 15-5

[0704] To a 4 mL reaction vial, compound 15-4 (50 mg, 271.49 μmol) and compound 1-7 (169.33 mg, 271.49 μmol) were added and dissolved in dry DMF (2 mL). HATU (119.29 mg, 352.94 μmol) and DIPEA (105.07 mg, 814.48 μmol) were added with stirring. After addition, the reaction was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, compound 15-5 (50 mg, 23.32% yield) was obtained directly by MPLC. LCMS (E+) m / z: 790.8 [M+H] + .

[0705] Step 4. Synthesis of compound 15

[0706] To a 4 mL reaction vial, compound 15-5 (50 mg, 63.30 μmol) was added and dissolved in methanol (1 mL). 1 M sodium hydroxide solution (1 mL) was then added. After complete addition, the reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, compound 15 (40 mg, 77.92% yield) was obtained directly by MPLC.

[0707] LCMS (E+) m / z: 776.8 [M+H] + .

[0708] 1H NMR(400MHz, DMSO-d6)δ8.49(d,J=8.3Hz,1H),8.30–8.25(m,1H),8.22(t,J=5.8Hz,1H),7.81–7.73(m,1H),7.5 7–7.49(m,2H),7.45(d,J=8.3Hz,2H),7.40(d,J=8.2Hz,2H),7.33(d,J=8.0Hz,1H),7.06(d,J=8.1Hz,1H),6.52 (dd,J=8.1,3.4Hz,1H),5.30(q,J=7.5Hz,1H),4.37–4.28(m,2H),3.96–3.90(m,2H),3.76(d,J=5.7Hz,2H),3.6 9(dd,J=5.9,3.6Hz,2H),3.63–3.45(m,11H),3.36(dd,J=5.6,4.3Hz,2H),2.87–2.77(m,4H),2.58–2.50(m,3H).

[0709] Example 16

[0710] Synthesis of compound 16

[0711] Step 1. Synthesis of compound 16-2

[0712] Under nitrogen, a 250 mL three-necked flask was charged with 16-1 (5.00 g, 18.30 mmol), methyl acrylate (1.89 g, 21.96 mmol), palladium acetate (414.53 mg, 1.83 mmol), tris(o-methylphenyl)phosphine (85.22 mg, 0.28 mmol), diisopropylethylamine (23.65 g, 183.00 mmol), and N,N-dimethylformamide (50 mL). The system was stirred at 100°C overnight and monitored by LC-MS. After completion of the reaction, the product was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The product was purified by column chromatography (petroleum ether:ethyl acetate = 5:1) to afford intermediate 16-2 (2.40 g, 47.15% yield). LCMS (E+) m / z: 279.2 [M+H] + .

[0713] Step 2. Synthesis of compound 16-3

[0714] Under nitrogen, palladium on carbon (100 mg) was slowly added to a solution of 16-2 (1.00 g, 3.59 mmol) in ethyl acetate (30 mL). The system was stirred at room temperature overnight and monitored by LC-MS. After the reaction, the reaction solution was filtered through celite and concentrated to obtain compound 16-3 (1.05 g, crude product), which was used directly in the next step. LCMS (E+) m / z: 281.1 [M+H] + .

[0715] Step 3. Synthesis of compound 16-4

[0716] Compound 16-3 (400 mg, 1.43 mmol) was added to a 20 mL reaction flask and dissolved in methanol (5 mL). Sodium hydroxide (171.60 mg, 4.29 mmol) was then added. The reaction was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, the reaction solution was directly analyzed by MPLC to yield compound 16-4 (370.00 mg, 97.36% yield). LCMS (E-) m / z: 265.2 [MH] + .

[0717] Step 4. Synthesis of Intermediate 16-5

[0718] Compound 16-4 (118 mg, 0.44 mmol) was added to a 20 mL reaction flask and dissolved in DMF (5 mL). Compound 1-7 (305.27 mg, 0.49 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (250.95 mg, 0.66 mmol), and N,N-diisopropylethylamine (284.33 mg, 2.2 mmol) were added while stirring at room temperature. The reaction mixture was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, the reaction solution was directly analyzed by MPLC to yield compound 16-5 (230 mg, 60.00% yield). LCMS (E+) m / z: 872.4 [M+H] + .

[0719] Step 5. Synthesis of compound 16-6

[0720] Compound 16-5 (230 mg, 0.26 mmol) was added to a 4 mL reaction flask and dissolved in 4 M hydrochloric acid in dioxane (1 mL) and dioxane (1 mL). The reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, the reaction solution was directly concentrated to afford product 16-6 (190 mg, 93.32% yield). LCMS (E+) m / z: 772.3 [M+H] + .

[0721] Step 6. Synthesis of compound 16

[0722] Compound 16-6 (190 mg, 0.25 mmol) was added to a 4 mL reaction flask and dissolved in methanol (1 mL). 1 M sodium hydroxide solution (1 mL) was then added. The reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, compound 16 (140 mg, 74.07% yield) was obtained directly by MPLC. LCMS (E+) m / z: 758.3 [M+H] + . 1 H NMR (600MHz, DMSO-d6) δ9.71(s,1H),8.41(t,J=6.0Hz,1H),8.26(d,J=8.2Hz,1H),7.80(d,J=8.2Hz,1H),7.51(dt,J= 18.0,7.0Hz,2H),7.42(d,J=7.8Hz,2H),7.30(d,J=7.6Hz,3H),7.24(t,J=7.7Hz,1H),7.03(d,J=8.0Hz,1H),6.35(d,J =7.3Hz,1H),6.24(d,J=8.1Hz,1H),5.82(s,2H),5.09(d,J=6.6Hz,1H),4.32(t,J=4.5Hz,2H),3.92(t,J=4.5Hz,2H), 3.70(dt,J=15.7,5.1Hz,4H),3.61–3.49(m,14H),2.76(t,J=8.0Hz,2H),2.53(d,J=6.4Hz,2H),2.43(d,J=5.5Hz,2H).

[0723] Example 17

[0724] Synthesis of compound 17

[0725] Step 1. Synthesis of compound 17-2

[0726] Compound 17-1 (800.00 mg, 3.41 mmol), triethylamine (516.06 mg, 5.10 mmol), and dichloromethane (3 mL) were added to a 10 mL round-bottom flask. After the system was cooled to 0°C, methylsulfonyl chloride (467.30 mg, 4.08 mmol) was added dropwise. The system was stirred at room temperature overnight and monitored by LC-MS. After the reaction, the mixture was washed twice with 1N HCl aqueous solution, washed with saturated sodium bicarbonate aqueous solution, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain intermediate compound 17-2 (783.12 mg, 73.56% yield). LCMS (E+) m / z: 314.1 [M+H] + .

[0727] Step 2. Synthesis of compound 17-3

[0728] At room temperature, tert-butyl (4-methylpyridin-2-yl)carbamate (400.00 mg, 1.92 mmol) was dissolved in 4 mL of N,N-dimethylformamide in a 25 mL round-bottom flask. Cesium carbonate (3.13 g, 9.61 mmol) was added and the atmosphere was replaced with nitrogen. Compound 17-2 (662.52 mg, 2.12 mmol) was dissolved in 4 mL of N,N-dimethylformamide and added dropwise to the above system. After the addition was complete, the system was stirred at 70°C for 3 hours and monitored by LC-MS. After the reaction was completed, the mixture was washed with ethyl acetate and saturated brine, the organic phases were combined, dried, concentrated, and MPLC was used to obtain compound 17-3 (746.59 mg, 82.98% yield). LCMS (E+) m / z: 426.2 [M+H] + .

[0729] Step 3. Synthesis of compound 17-4

[0730] Compound 17-3 (500.00 mg, 1.18 mmol) was added to a 20 mL reaction flask, dissolved in methanol (2.5 mL), and then palladium on carbon (400.00 mg) was added. The system was purged with hydrogen, and the reaction was stirred at room temperature overnight and monitored by LC-MS. After completion of the reaction, the reaction solution was directly analyzed by MPLC to obtain compound 17-4 (313.51 mg, 91.62% yield). LCMS (E+) m / z: 292.0 [M+H] + .

[0731] Step 4. Synthesis of compound 17-5

[0732] At room temperature, compound 17-4 (300.00 mg, 1.03 mmol), potassium carbonate (284.96 mg, 2.06 mmol), and N,N-dimethylformamide (3 mL) were added to a 10 mL round-bottom flask. Methyl bromopropionate (206.60 mg, 1.24 mmol) was dissolved in 2 mL of N,N-dimethylformamide and added dropwise to the above system. After completion, the system was stirred at room temperature for 3 hours and monitored by LC-MS. After completion of the reaction, the mixture was washed with ethyl acetate and saturated brine, and the organic phases were combined, dried, and concentrated. Compound 17-5 (282.51 mg, 72.63% yield) was obtained using MPLC. LCMS (E+) m / z: 378.3 [M+H] + .

[0733] Step 5. Synthesis of compound 17-6

[0734] Compound 17-5 (250.00 mg, 0.67 mmol) and 5 mL of methanol were added to a 25 mL round-bottom flask. 5 mL of 2N aqueous sodium hydroxide solution was added dropwise. The mixture was stirred at room temperature for 1 hour and monitored by LC-MS. After the reaction, the pH of the reaction solution was adjusted to a weakly acidic state. MPLC analysis yielded compound 17-6 (229.63 mg, 95.67% yield). LCMS (E-) m / z: 362.2 [MH] + .

[0735] Step 6. Synthesis of compound 17-7

[0736] At room temperature, compound 17-6 (100.00 mg, 0.28 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (114.21 mg, 0.30 mmol), N,N-diisopropylethylamine (161.75 mg, 1.25 mmol), and N,N-dimethylformamide (3 mL) were added to a 10 mL reaction vial. Intermediate 1-5 (141.74 mg, 0.25 mmol) was dissolved in 2 mL of N,N-dimethylformamide and added dropwise to the above system. After the addition, the mixture was stirred at room temperature for 3 hours and monitored by LC-MS. After completion of the reaction, compound 17-7 (169.65 mg, 67.16% yield) was obtained directly by MPLC. LCMS (E+) m / z: 913.4 [M+H] + .

[0737] Step 7. Synthesis of compound 17-8

[0738] Compound 17-7 (169.65 mg, 0.19 mmol) was added to a 4 mL reaction vial at room temperature, followed by a 4 M solution of hydrogen chloride in dioxane (2 mL). The mixture was stirred for 2 hours and monitored by LC-MS. After completion of the reaction, the reaction solution was directly concentrated to afford compound 17-8 (145.12 mg, 96.21% yield). LCMS (E-) m / z: 810.2 [MH]+.

[0739] Step 8. Synthesis of compound 17

[0740] Compound 17-8 (54.82 mg, 0.07 mmol) and 1 mL of methanol were added to a 4 mL reaction flask. 1 mL of 2N sodium hydroxide solution was added dropwise. The mixture was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, the reaction mixture was analyzed by MPLC to yield 17 (49.20 mg, 91.34% yield).

[0741] LCMS (E-) m / z: 796.1 [MH] + .

[0742] 1 H NMR (400MHz, DMSO-d6) δ9.43(d,J=7.5Hz,1H),8.25(d,J=7.8Hz,1H),7.80(d,J=7.8Hz,1H),7.74(d,J=5.1Hz,1H),7.51(q,J=7.1Hz ,2H),7.44(d,J=8.0Hz,2H),7.30(dd,J=7.9,3.2Hz,3H),7.07–6.97(m,2H),6.32–6.15(m,2H),5.09(q,J=5.7Hz,1H),4.35–4.26(m, 2H),3.95–3.88(m,2H),3.71–3.67(m,2H),3.62–3.45(m,14H),3.26–3.21(m,1H),3.05(dt,J=13.3,6.8Hz,1H),2.99(s,1H),2.74(s ,1H),2.61(ddd,J=24.8,11.1,4.6Hz,4H),2.43–2.33(m,5H),2.28(t,J=6.5Hz,2H),1.87(dt,J=8.5,4.5Hz,1H),1.49–1.10(m,3H).

[0743] Example 18

[0744] Synthesis of compound 18

[0745] Step 1. Synthesis of compound 18-2

[0746] Compound 18-1 (800.00 mg, 3.41 mmol), triethylamine (516.06 mg, 5.10 mmol), and dichloromethane (6 mL) were added to a 25 mL round-bottom flask. After the system was cooled to 0°C, methylsulfonyl chloride (467.30 mg, 4.08 mmol) was added dropwise. The system was stirred at room temperature overnight and monitored by LC-MS. After completion of the reaction, the product was washed twice with 1N HCl, saturated sodium bicarbonate, and saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain intermediate compound 18-2 (783.12 mg, 73.56% yield). LCMS (E+) m / z: 314.1 [M+H] + .

[0747] Step 2. Synthesis of compound 18-3

[0748] At room temperature, tert-butyl (4-methylpyridin-2-yl)carbamate (400.00 mg, 1.92 mmol) was dissolved in 4 mL of N,N-dimethylformamide in a 25 mL round-bottom flask. Cesium carbonate (3.13 g, 9.61 mmol) was added and the atmosphere was replaced with nitrogen. Compound 18-2 (662.52 mg, 2.12 mmol) was dissolved in 4 mL of N,N-dimethylformamide and added dropwise to the above system. After the addition was complete, the system was stirred at 70°C for 3 hours and monitored by LC-MS. After the reaction was completed, the mixture was extracted with ethyl acetate and saturated brine, the organic phases were combined, dried, concentrated, and MPLC was used to obtain compound 18-3 (746.59 mg, 82.98% yield). LCMS (E+) m / z: 426.2 [M+H] + .

[0749] Step 3. Synthesis of compound 18-4

[0750] Compound 18-3 (500.00 mg, 1.18 mmol) was added to a 20 mL reaction flask, dissolved in methanol (2.5 mL), and then palladium on carbon (400 mg). The system was purged with hydrogen, and the reaction was stirred at room temperature overnight and monitored by LC-MS. After completion of the reaction, the reaction solution was directly analyzed by MPLC to obtain compound 18-4 (313.51 mg, 91.62% yield). LCMS (E+) m / z: 292.0 [M+H] + .

[0751] Step 4. Synthesis of compound 18-5

[0752] At room temperature, compound 18-4 (300.00 mg, 1.03 mmol), triethylamine (208 mg, 2.06 mmol), and N,N-dimethylformamide (3 mL) were added to a 10 mL round-bottom flask. Methyl bromoacetate (236.6 mg, 1.55 mmol) was dissolved in 2 mL of N,N-dimethylformamide and added dropwise to the above system. After completion, the system was stirred at room temperature for 3 hours and monitored by LC-MS. After completion of the reaction, the mixture was extracted with ethyl acetate and saturated brine. The organic phases were combined, dried, and concentrated. Compound 18-5 (287 mg, 76.82% yield) was obtained using MPLC. LCMS (E+) m / z: 364.5 [M+H] + .

[0753] Step 5. Synthesis of compound 18-6

[0754] Compound 18-5 (250.00 mg, 0.69 mmol) and 5 mL of methanol were added to a 25 mL round-bottom flask. 5 mL of 2N aqueous sodium hydroxide solution was added dropwise. The mixture was stirred at room temperature for 1 hour and monitored by LC-MS. After the reaction, the pH of the reaction solution was adjusted to a weakly acidic state. MPLC analysis yielded compound 18-6 (239.58 mg, 95.31% yield). LCMS (E+) m / z: 350.4 [M+H] + .

[0755] Step 6. Synthesis of compound 18-7

[0756] At room temperature, compound 18-6 (100.00 mg, 0.29 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (130.74 mg, 0.34 mmol), N,N-diisopropylethylamine (185.16 mg, 1.43 mmol), and N,N-dimethylformamide (3 mL) were added to a 10 mL reaction vial. Intermediate 1-5 (147.50 mg, 0.26 mmol) was dissolved in 2 mL of N,N-dimethylformamide and added dropwise to the above system. After the addition, the mixture was stirred at room temperature for 3 hours and monitored by LC-MS. After completion of the reaction, compound 18-7 (190.26 mg, 74.07% yield) was obtained directly by MPLC. LCMS (E+) m / z: 898.5 [M+H] + .

[0757] Step 7. Synthesis of compound 18-8

[0758] Compound 18-7 (190.00 mg, 0.21 mmol) was added to a 4 mL reaction vial at room temperature, followed by a 4 M solution of hydrogen chloride in dioxane (2 mL). The mixture was stirred for 2 hours and monitored by LC-MS. After completion of the reaction, the reaction solution was directly concentrated to afford compound 18-8 (158.13 mg, 93.32% yield). LCMS (E+) m / z: 798.4 [M+H]+.

[0759] Step 8. Synthesis of compound 18

[0760] Compound 18-8 (17.73 mg, 0.02 mmol) and 1 mL of methanol were added to a 4 mL reaction flask. 1 mL of 2N sodium hydroxide solution was added dropwise. The mixture was stirred at room temperature for 1 hour and monitored by LC-MS. After the reaction, MPLC of the reaction mixture yielded compound 18 (16.60 mg, 95.31% yield).

[0761] LCMS (E-) m / z: 782.2 [MH] + .

[0762] 1 H NMR (600MHz, DMSO-d6) δ8.27(dd,J=8.0,1.6Hz,1H),7.83–7.68(m,2H),7.52(dddd,J=17.9,8.2,6.7,1.5 Hz,2H),7.48–7.45(m,2H),7.39(d,J=7.9Hz,2H),7.33(d,J=7.8Hz,1H),7.06(d,J=8.0Hz,1H),6.49(s,1H ),6.33–6.21(m,2H),5.32(q,J=8.0Hz,1H),4.33(dd,J=5.7,3.5Hz,2H),3.95–3.90(m,2H),3.69(dd,J=5. 8,3.7Hz,2H),3.61–3.45(m,21H),2.85(qd,J=15.8,7.1Hz,2H),2.04–1.87(m,2H),1.24(d,J=5.5Hz,7H).

[0763] Using conditions similar to those in the above examples, the compounds listed in Table 1 were prepared. The structural characterization data of these compounds are listed in Table 1.

[0764] Table 1

[0765] Example 19

[0766] Synthesis of compound 58

[0767] Step 1. Synthesis of compound 58-2

[0768] Compound 58-1 (5.00 g, 24.84 mmol) was added to the reaction flask and dissolved in dichloromethane (50 mL). Dess-Martin periodinane (21.07 g, 49.69 mmol) was added with stirring. After the addition, the system was stirred at 25°C for 1 hour and monitored by TLC. The reaction system was quenched with saturated sodium bicarbonate solution (200 mL), extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase was concentrated to obtain the crude product 58-2 (2.40 g). The crude product was used directly in the next reaction. LCMS (E+) m / z: 200.25 [M+H] + .

[0769] Step 2. Synthesis of compound 58-3

[0770] The crude product 58-2 (2.39 g, 22.08 mmol) was added to the reaction flask and dissolved in DCM (50 mL). The system was cooled to 0°C under nitrogen protection, and sodium triacetoxyborohydride (6.38 g, 30.11 mmol) was added in batches with stirring. After the addition, the mixture was stirred for another 4 hours and the reaction was monitored by LCMS. After completion, the reaction was quenched with saturated sodium bicarbonate solution (200 mL), extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (PE / EA = 1:3) to give compound 58-3 (3.20 g, 9.88 mmol, yield 49.23%, purity 90%) as a white oil. LCMS (E+) m / z: 292.19 [M+H] + .

[0771] Step 3. Synthesis of compound 58-4

[0772] Compound 58-3 (3.00 g, 10.30 mmol) was added to the reaction flask and dissolved in dichloromethane (30 mL). The system was cooled to 0°C under nitrogen, and benzoyloxycarbonyl succinimide (7.03 g, 41.18 mmol) and N,N-diisopropylethylamine (3.99 g, 30.89 mmol, 5.38 mL) were slowly added with stirring. After addition, the mixture was stirred at 0°C for 2 hours and monitored by LCMS. After completion, water (200 ml) was added to quench the reaction, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (PE / EA = 1:1) to obtain compound 58-4 (2.00 g, 4.47 mmol, 43.37% yield, 95% purity) as a white solid. LCMS (E+) m / z: 426.23 [M+H] + .

[0773] Step 4. Synthesis of compound 58-5

[0774] Compound 58-4 (2.00 g, 4.70 mmol) was added to the reaction flask, followed by a mixed solvent of trifluoroacetic acid (5.00 ml) and dichloromethane (10.00 mL). The mixture was stirred at 25°C for 1 hour, and the reaction was monitored by LCMS. After completion of the reaction, the system was concentrated to obtain the crude product. MPLC analysis yielded compound 58-5 (1.10 g, 3.21 mmol, 68.33% yield, 95% purity) as a white solid. LCMS (E+) m / z: 326.18 [M+H] + .

[0775] Step 5. Synthesis of compound 58-6

[0776] Compound 58-5 (800.00 mg, 2.33 mmol) was added to the reaction flask and dissolved in DMF (10 mL). The system was cooled to 0-5°C under nitrogen protection, and methyl 2-bromoacetate (413.00 mg, 2.70 mmol) and DBU (1.12 g, 7.38 mmol) were added in sequence. After the addition, the reaction was stirred at 0°C for 3 hours and monitored by LCMS. After the reaction was completed, the reaction system was not treated and was directly used in the next step. LCMS (E+) m / z: 398.20 [M+H] + .

[0777] Step 6. Synthesis of compound 58-7

[0778] To the above reaction system, NaOH (301.00 mg, 7.55 mmol), MeOH (10.0 mL), and water (10.0 mL) were added. After addition, the mixture was stirred at 0°C for 0.5 hours and the reaction was monitored by LCMS. After completion of the reaction, the crude product was purified by MPLC to afford compound 58-7 (650.00 mg, 1.70 mmol, 67.38% yield) as a yellow solid. LCMS (E+) m / z: 384.18 [M+H] + .

[0779] Step 7. Synthesis of compound 58-8

[0780] Compound 58-7 (150.0 mg, 391.20 μmol) and Pd / C (20.00 mg, 391.20 μmol) were dissolved in MeOH (3 mL) and replaced with hydrogen three times. After replacement, the reaction was stirred at 25°C under a hydrogen atmosphere for half an hour and monitored by LCMS. After completion of the reaction, the reaction system was filtered through celite, the filter cake was rinsed with MeOH (3*20) and the filtrate was collected. The filtrate was concentrated under reduced pressure to obtain a white oil compound 58-8 (120.00 mg, 385.07 μmol, yield 98.43%, purity 80%). LCMS (E+) m / z: 250.15 [M+H] + .

[0781] Step 8. Synthesis of compound 58-9

[0782] Compound 58-8 (120.00 mg, 385.07 μmol) was added to the reaction flask and dissolved in dichloromethane (5 mL). 58-8' (136.00 mg, 240.67 μmol), DIPEA (187.00 mg, 1.44 μmol), EDCI (110.00 mg, 577.60 μmol), and HOBt (78.00 mg, 577.60 μmol) were added sequentially with stirring. After the addition, the system was stirred at 25°C for 3 hours and the reaction was monitored by LCMS. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The crude product was purified by MPLC to obtain compound 58-9 (130.00 mg, 154.77 μmol, 32.16% yield, 95% purity) as a white oil. LCMS (E+) m / z: 798.41 [M+H] + .

[0783] Step 9. Synthesis of Compound 58

[0784] Compound 58-9 (130.00 mg, 154.77 μmol) was added to a reaction flask, followed by tetrahydrofuran (1 mL), methanol (1 mL), and water (1 mL). After stirring and dissolving, sodium hydroxide (31.00 mg, 773.85 μmol) was added. After addition, the system was stirred at 25°C for half an hour. The reaction was monitored by LCMS. After completion of the reaction, the system was concentrated under reduced pressure to obtain a solid crude product. The crude product was purified by MPLC to obtain compound 58 (74.50 mg, 93.80 μmol, yield 62.37%, purity 98.7%) as a white solid.

[0785] LCMS (E+) m / z: 784.40 [M+H] + .

[0786] 1H NMR(400MHz, DMSO-d6)δ8.25(dd,J=7.6,1.8Hz,1H),7.83–7.71(m,2H),7.55–7.45(m,2H),7.44–7.38(m,2H),7.30(d d,J=8.1,6.8Hz,4H),7.02(d,J=8.1Hz,1H),6.36(t,J=5.6Hz,1H),6.26(d,J=4.2Hz,2H),5.10(dt,J=8.2,5.6Hz,1H) ,4.31(dd,J=5.7,3.5Hz,2H),3.97–3.87(m,2H),3.69(dd,J=5.9,3.6Hz,2H),3.62–3.43(m,16H),3.39–3.34(m,2H), 3.15(q,J=6.7Hz,2H),3.01(s,2H),2.88(td,J=6.7,3.6Hz,2H),2.60–2.41(m,1H)2.09(s,3H),1.75(q,J=7.1Hz,2H).

[0787] Example 20

[0788] Synthesis of compound 59

[0789] Step 1. Synthesis of compound 59-2

[0790] Under nitrogen protection, compounds 59-1 (2.00 g, 10.52 mmol) and 59-1' (5.69 g, 52.60 mmol) were dissolved in a mixed solvent of tetrahydrofuran (15 mL) and dichloroethane (15 mL). After stirring at room temperature for half an hour, sodium triacetoxyborohydride (11.15 g, 52.60 mmol) was added portionwise. After addition, the system was heated to 60°C and reacted overnight, monitored by LC-MS. After completion of the reaction, water (10 mL) was added to quench the reaction, and compound 59-2 (2.10 g, 7.44 mmol, 70.76% yield) was obtained by reverse phase reaction. LCMS (E+) m / z: 283.1 [M+H] + .

[0791] Step 2. Synthesis of compound 59-3

[0792] Under nitrogen, compound 59-2 (2.00 g, 10.52 mmol) was dissolved in methanol (30 mL). Pd / C (300.0 mg, 20% w) was added at room temperature, and the system was replaced with hydrogen three times. After addition, the system was heated to 40°C and allowed to react overnight, monitored by LC-MS. After completion of the reaction, the mixture was filtered and the methanol was concentrated to afford compound 59-3 (1.50 g, 8.20 mmol, 77.87% yield). LCMS (E+) m / z: 183.1 [M+H] + .

[0793] Step 3. Synthesis of compound 59-4

[0794] Compound 59-3 (1.40 g, 7.65 mmol) and triethylamine (1.54 g, 15.30 mmol) were dissolved in dichloromethane (10 mL). The mixture was cooled to 0-5°C under nitrogen and methylsulfonyl chloride (1.05 g, 9.17 mmol) was slowly added. The reaction was allowed to react for half an hour. LC-MS monitoring was performed. After the reaction was completed, water (10 mL) was added to quench the reaction. Compound 59-4 (160.00 mg, 0.60 mmol, 5.70% yield) was obtained by reverse phase chromatography. LCMS (E+) m / z: 271.0 [M+H] +

[0795] Step 4. Synthesis of compound 59-5

[0796] Under nitrogen, compounds 59-4 (160.00 mg, 0.60 mmol) and 59-4' (128.90 mg, 0.89 mmol) were dissolved in DMF (10 mL), and potassium carbonate (182.20 mg, 1.32 mmol) was added. The system was heated to 60°C and reacted overnight, monitored by LC-MS. After completion of the reaction, compound 59-5 (40.00 mg, 0.12 mmol, 20.10% yield) was obtained by reverse phase purification. LCMS (E+) m / z: 320.2 [M+H] +

[0797] Step 5. Synthesis of compound 59-6

[0798] Compound 59-5 (30.00 mg, 0.09 mmol) was dissolved in methanol (2 mL) at room temperature, and 1 M sodium hydroxide solution (2 mL) was added with stirring. The system was heated to 40°C and reacted for 1 hour, monitored by LC-MS. After the reaction, compound 59-6 (22.00 mg, 0.08 mmol, 93.41% yield) was obtained by reverse phase reaction. LCMS (E+) m / z: 264.1 [M+H] +

[0799] Step 6. Synthesis of compound 59-7

[0800] To a 4 mL reaction vial, compound 59-6 (20.00 mg, 0.06 mmol) and 59-6' (24.83 mg, 0.04 mmol) were added sequentially and dissolved in N,N'-dimethylformamide (2 mL). N,N-diisopropylethylamine (16.18 mg, 0.13 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (27.36 mg, 0.07 mmol) were added to the system with stirring. The reaction was allowed to react at room temperature for 1 hour, monitored by LC-MS. After completion of the reaction, the reaction solution was purified by MPLC to yield compound 59-7 (45.00 mg, 0.05 mmol, 76.13% yield). LCMS (E+) m / z: 812.3 [M+H] + .

[0801] Step 7. Synthesis of Compound 59

[0802] To a 10 mL reaction flask, compound 59-7 (45.00 mg, 0.55 mmol) was added and dissolved in methanol (2.5 mL). 1 M sodium hydroxide solution (2.5 mL) was then added. After complete addition, the mixture was heated to 40°C and stirred for 1 hour, monitored by LC-MS. After completion of the reaction, compound 59 (20.00 mg, 0.02 mmol, 41.16% yield) was obtained directly by MPLC.

[0803] LCMS (E+) m / z: 798.7 [M+H] + .

[0804] 1 H NMR (600MHz, DMSO-d6) δ9.21 (s, 1H), 8.49–8.13 (m, 1H), 7.82 (d, J = 7.5Hz, 1H), 7.74 (d, J=8.3Hz,1H),7.63–7.37(m,6H),7.31(d,J=7.8Hz,1H),7.04(d,J=8.0Hz,1H),6.79–6.5 3(m,2H),5.47–5.23(m,1H),4.32(t,J=4.6Hz,2H),4.25–3.80(m,4H),3.69(dd,J=5.9,3 .7Hz,2H),3.64–3.40(m,14H),2.93–2.68(m,6H),2.32–2.09(m,5H),1.33–1.07(m,5H).

[0805] Example 21

[0806] Synthesis of compound 60

[0807] Step 1. Synthesis of compound 60-2

[0808] Compound 60-1 (1.00 g, 8.06 mmol), 60-1′ (1.75 g, 12.09 mmol), potassium carbonate (2.34 g, 16.92 mmol), and tetrabutylammonium iodide (0.60 g, 1.61 mmol) were added to a reaction flask and dissolved in DMSO (10 mL). The system was heated to 100°C overnight and monitored by LC-MS. After completion of the reaction, compound 60-2 (350.0 mg, 1.29 mmol, 16.03% yield) was obtained by reverse phase chromatography. LCMS (E+) m / z: 250.4 [M+H] + .

[0809] Step 2. Synthesis of compound 60-3

[0810] Compounds 60-2 (320.00 mg, 1.28 mmol) and 60-2' (694.03 mg, 6.42 mmol) were dissolved in tetrahydrofuran (10 mL) and dichloroethane (10 mL). After stirring at room temperature for 0.5 hours, sodium triacetoxyborohydride (1.36 g, 6.42 mmol) was added. The reaction was allowed to react at room temperature for 3 hours, monitored by LC-MS. After completion of the reaction, compound 60-3 (238.00 mg, 0.64 mmol, 49.96% yield) was obtained by reverse phase purification. LCMS (E+) m / z: 342.1 [M+H] + .

[0811] Step 3. Synthesis of compound 60-4

[0812] Compound 60-3 (230.00 mg, 0.67 mmol) was added to the reaction flask and dissolved in methanol (2.5 mL). 1 M aqueous sodium hydroxide solution (2.5 mL) was added at room temperature. After addition, the temperature was raised to 50°C and the reaction was allowed to react for 6 hours, monitored by LC-MS. After completion of the reaction, compound 60-4 (120.00 mg, 0.40 mmol, 59.31% yield) was obtained by reverse phase purification. LCMS (E+) m / z: 286.1 [M+H] + .

[0813] Step 4. Synthesis of compound 60-5

[0814] Compounds 60-4 (100.00 mg, 0.35 mmol) and 60-4' (198.59 mg, 0.35 mmol) were added to a 4 mL reaction vial and dissolved in N,N'-dimethylformamide (2 mL). N,N-diisopropylethylamine (68.00 mg, 0.60 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (160.56 mg, 0.42 mmol) were added with stirring. The reaction was allowed to react at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, the reaction solution was purified by MPLC to yield compound 60-5 (160.00 mg, 0.20 mmol, 55.68% yield). LCMS (E+) m / z: 834.7 [M+H] + .

[0815] Step 5. Synthesis of compound 60

[0816] To a 10 mL reaction flask, compound 60-5 (160.00 mg, 0.20 mmol) was added and dissolved in methanol (2.5 mL). 1 M sodium hydroxide solution (2.5 mL) was then added. After complete addition, the reaction was stirred at 40°C for 1 hour and monitored by LC-MS. After completion of the reaction, compound 60 (120.00 mg, 0.14 mmol, 72.49% yield) was obtained directly by MPLC.

[0817] LCMS (E+) m / z: 820.3 [M+H] + .

[0818] 1 H NMR(600MHz,DMSO-d6)δ8.57(d,J=8.5Hz,1H),8.27(dd,J=7.5,2.0Hz,1H),7.81–7.71(m,2H),7.58–7.47(m,2H) ,7.43(d,J=8.1Hz,2H),7.37(d,J=7.9Hz,2H),7.32(d,J=7.8Hz,1H),7.16(d,J=8.5Hz,2H),7.05(d,J=8.0Hz,1H ),6.73–6.58(m,3H),6.54(d,J=6.2Hz,1H),5.30(q,J=7.5Hz,1H),4.47–4.27(m,4H),4.02–3.87(m,4H),3.69(d d,J=5.9,3.7Hz,2H),3.62–3.48(m,12H),3.36(d,J=4.9Hz,2H),2.99(s,3H),2.80(d,J=7.3Hz,2H),2.21(s,3H).

[0819] Example 22

[0820] Synthesis of compound 1008A

[0821] Step 1. Synthesis of compound 1008-2

[0822] Under nitrogen, compound 1008-1 (2.00 g, 4.75 mmol), 1008-1' (2.20 g, 5.69 mmol), and potassium acetate (654.83 mg, 4.75 mmol) were dissolved in N,N-dimethylformamide (40 mL). The mixture was stirred at 90°C for 16 hours and monitored by LC-MS. After the reaction was completed, the mixture was cooled to room temperature, water (50 mL) was added, and the mixture was extracted with ethyl acetate. The mixture was washed with saturated brine and dried over anhydrous sodium sulfate. Reverse phase chromatography was performed to obtain compound 1008-2 (2.60 g, 3.89 mmol, 81.88% yield) as a yellow solid. LCMS (E+) m / z: 636.31 [M+H] + .

[0823] Step 2. Synthesis of Intermediate 1008-3

[0824] Compound 1008-2 (2.20 g, 3.46 mmol) was added to a mixture of dichloromethane (15 mL) and trifluoroacetic acid (5 mL) at 0°C. The mixture was stirred at 0°C for 1 hour. The reaction was monitored by LC-MS. After completion, the reaction was spin-dried and the crude product was purified by reverse phase chromatography to obtain intermediate 1008-3 (1.70 g, 3.01 mmol, 87.10% yield) as a yellow oil. LCMS (E+) m / z: 536.26 [M+H] + .

[0825] Step 3. Synthesis of Compound 1008-4

[0826] Compound N-Boc glycine (302.63 mg, 0.53 mmol) was added to a 25 mL reaction vial at room temperature and dissolved in DMF (5 mL). Compound 1008-3 (110.25 mg, 0.63 mmol), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (220.40 mg, 0.58 mmol), and N,N-diisopropylethylamine (135.45 mg, 1.05 mmol) were added sequentially with stirring. The reaction was allowed to proceed for 30 minutes, monitored by LC-MS. After completion of the reaction, compound 1008-4 (352.92 mg, 0.51 mmol, 96.23% yield) was obtained by MPLC purification. LCMS (E+) m / z: 693.6 [M+H] + .

[0827] Step 4. Synthesis of Compound 1008-5

[0828] Compound 1008-4 (352.92 mg, 0.51 mmol) was added to a 25 mL single-necked vial at room temperature and dissolved in dichloromethane (5 mL). Trifluoroacetic acid (2 mL) was then added while stirring in an ice-water bath at 0°C. The reaction was allowed to react at room temperature for 10 minutes, monitored by LC-MS. After completion of the reaction, intermediate 1008-5 (301.00 mg, 0.51 mmol, 99.70% yield) was obtained by MPLC purification. LCMS (E+) m / z: 593.9 [M+H] + .

[0829] Step 5. Synthesis of Compound 1008-6A

[0830] To a 4 mL reaction vial were added compound 8-5A (50.00 mg, 0.24 mmol, a chirally pure intermediate from the synthesis of compound 8), 1008-5 (100.00 mg, 0.17 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (95.00 mg, 0.23 mmol), and N,N-diisopropylethylamine (100 μL, 0.60 mmol). The mixture was reacted at room temperature for 1 hour until the starting material disappeared as determined by LC-MS. The reaction solution was directly analyzed by MPLC to yield compound 1008-6A (70.00 mg, 87.84 μmol, 51.73% yield). LCMS (E+) m / z: 797.6 [M+H] + .

[0831] Step 6. Synthesis of Compound 1008A

[0832] Compound 1008-6A (70.00 mg, 87.84 μmol) was added to a 25 mL reaction flask, followed by dissolution in methanol (3 mL). Sodium hydroxide solution (2N, 3 mL) was then added dropwise. The reaction was stirred at 25°C for 1 hour. After the reaction, the methanol was concentrated, and the system was purified by MPLC to yield compound 1008A (50.00 mg, 63.87 μmol, 72.56% yield).

[0833] LCMS (E+) m / z: 783.4 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ9.67(s,1H),8.36(t,J=6.0Hz,1H),8.26(dd,J=7.8,1.8Hz,1H),7.78(dd,J=7.8,1.7Hz,1H),7.50(dddd,J=15.3,8.3,6.8,1.6Hz ,2H),7.45–7.38(m,3H),7.30(dd,J=8.0,3.5Hz,3H),7.02(d,J=8.0Hz,1H), 6.81(d,J=2.5Hz,1H),6.32(d,J=5.0Hz,1H),5.09(d,J=6.6Hz,1H),4.32(dd ,J=5.8,3.5Hz,2H),4.12(d,J=2.4Hz,2H),4.08(dd,J=10.7,2.9Hz,1H),3.9 2(dd,J=5.7,3.5Hz,2H),3.84(dd,J=10.7,5.6Hz,1H),3.79–3.63(m,4H),3. 59(dd,J=5.9,3.7Hz,2H),3.56–3.49(m,9H),3.41(t,J=2.4Hz,2H),2.46(d, J=5.7Hz,2H),2.39–2.29(m,2H),2.03(s,3H),1.74(dt,J=12.9,7.0Hz,2H).

[0834] Example 23

[0835] Synthesis of compound 1009

[0836] Step 1. Synthesis of Compound 1009-1A: Compound 8-4 (1.79 g, 5.59 mmol, derived from the synthesis of compound 8) was subjected to chiral separation by SFC to afford compounds 1009-1A (810.00 mg, 2.53 mmol, SFC retention time 1.35 min) and 1009-1B (805.00 mg, 2.51 mmol, retention time 1.74 min). The resulting intermediate 1009-1A was determined to be the most active fragment by comparison with compound 8, and was therefore used in the next step.

[0837] Step 2. Synthesis of Compound 1009-2A

[0838] Compound 1009-1A (250.00 mg, 780.33 μmol) was added to a 4 mL reaction flask and dissolved in methanol (1 mL). 1 M sodium hydroxide solution (1 mL) was then added. After addition, the reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, compound 1009-2A (238.00 mg, 775.26 μmol, 99.35% yield) was obtained directly by MPLC. LCMS (E+) m / z: 307.1 [M+H]+

[0839] Step 1. Synthesis of compound 1009-3A

[0840] To a 4 mL reaction vial, compound 1009-2A (60.00 mg, 19.58 μmol) and compound 1008-5 (116.07 mg, 19.58 μmol) were added and dissolved in dry DMF (2 mL). HATU (89.28 mg, 23.49 μmol) and DIPEA (75.77 mg, 58.74 μmol) were added with stirring. After addition, the reaction was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, compound 1009-3A (95.00 mg, 10.78 μmol, 55.07% yield) was obtained directly by MPLC. LCMS (E+) m / z: 881.3 [M+H] + .

[0841] Step 2. Synthesis of Compound 1009-4A

[0842] Compound 1009-3A (95.00 mg, 10.78 μmol) was added to a 4 mL reaction vial and dissolved in 4 M dioxane hydrochloride (0.5 mL). The reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, compound 1009-4A (80.00 mg, 10.24 μmol, 95.03% yield) was obtained directly by MPLC. LCMS (E+) m / z: 791.5 [M+H] + .

[0843] Step 3. Synthesis of Compound 1009A

[0844] Compound 1009-4A (80.00 mg, 10.24 μmol) was added to a 4 mL reaction vial and dissolved in methanol (1 mL). 1 M sodium hydroxide solution (1 mL) was then added. The reaction was stirred at room temperature for 1 hour and monitored by LC-MS. After completion of the reaction, 1009A (50.00 mg, 6.52 μmol, 63.67% yield) was obtained directly by MPLC.

[0845] LCMS (E+) m / z: 767.7 [M+H]+ .

[0846] 1 H NMR (400MHz, DMSO-d6) δ9.70(d,J=7.7Hz,1H),8.40(t,J=6.0Hz,1H),8.26(dd,J=7.6,1.9Hz,1H),7.79(dd,J=7.7,1.8Hz,1H),7.5 8–7.37(m,5H),7.30(dd,J=8.0,2.4Hz,3H),7.03(d,J=8.0Hz,1H),6.58(d,J=2.0Hz,1H),6.21(d,J=5.4Hz,1H),5.08(dt,J=7.8,5 .4Hz,1H),4.32(dd,J=5.8,3.5Hz,2H),4.12(d,J=2.4Hz,2H),3.92(dd,J=5.7,3.4Hz,2H),3.84–3.48(m,15H),3.42(t,J=2.4Hz,1 H),3.05(dd,J=16.3,9.2Hz,1H),2.54–2.48(m,5H),2.41(d,J=5.4Hz,2H),2.29(q,J=6.8,6.4Hz,2H),1.77(qd,J=7.2,3.2Hz,2H).

[0847] Example 24

[0848] Synthesis of compound 1013

[0849] Step 1. Synthesis of compound 1013-2

[0850] At room temperature, 1,2-dichloroethane (50 mL) was added to a 250 mL single-necked flask, followed by compound 1013-1 (1.09 g, 5.16 mmol), and the mixture was stirred until fully dissolved. Compound 1013-1' (836.94 mg, 7.74 mmol) and tetraisopropyl titanate (2.20 g, 7.74 mmol) were slowly added to the above system. The system was stirred at room temperature for 30 minutes, and then sodium acetate borohydride (5.47 g, 25.80 mmol) was added. After the addition, the reaction system was stirred at 70°C for 2 hours. The reaction system was diluted with saturated sodium bicarbonate, extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, and the organic layer was concentrated. The crude product was purified by MPLC to obtain compound 1013-2 (696 mg, 2.29 mmol, 44.46% yield). LCMS (E+) m / z: 304.2 [M+H] + .

[0851] Step 2. Synthesis of Compound 1013-3

[0852] To a 50 mL single-necked flask, add intermediate 1013-2 (696.00 mg, 2.29 mmol), followed by dichloromethane (2 mL) and 4 M hydrochloric acid in dioxane (8 mL). Stir the mixture at room temperature for 2.5 hours, monitoring by LC-MS. After completion of the reaction, concentrate the mixture to afford crude compound 1013-3 (466 mg). This crude product was used directly in the next step.

[0853] Step 3. Synthesis of compound 1013-4

[0854] Intermediate 1013-3 (466 mg, 2.29 mmol) was added to a 25 mL single-necked flask and dissolved in DMF (7 mL). DBU (1.05 g, 6.88 mmol) and 1013-3′ (420.81 mg, 2.75 mmol) were then added sequentially. The reaction system was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, the reaction solution was directly analyzed by MPLC to yield compound 1013-4 (500.00 mg, 1.82 mmol, 79.21% yield). LCMS (E+) m / z: 276.2 [M+H] + .

[0855] Step 4. Synthesis of Compound 1013-5

[0856] Compound 1013-4 (500 mg, 1.82 mmol) was added to a 25 mL reaction flask and dissolved in anhydrous methanol (4.5 mL) and water (1.5 mL). Lithium hydroxide monohydrate (457.61 mg, 10.90 mmol) was then added. The reaction mixture was stirred at room temperature for 2 hours and monitored by LC-MS. After the reaction, the reaction solution was directly analyzed by MPLC to obtain compound 1013-5 (455.00 mg, 1.74 mmol, 95.88% yield). LCMS (E+) m / z: 262.2 [M+H] + .

[0857] Step 5. Synthesis of Compound 1013-6

[0858] To a 10 mL single-necked vial, intermediate 1013-5 (70.25 mg, 268.84 μmol), N,N-diisopropylethylamine (69.49 mg, 537.69 μmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (76.62 mg, 201.63 μmol), and DMF (1 mL) were added. The system was stirred at room temperature for 10 minutes, followed by the addition of compound 1008-3 (72.00 mg, 134.42 μmol). The reaction was allowed to react at room temperature for 2 hours until the starting material disappeared as determined by LC-MS. The reaction solution was directly subjected to MPLC to afford compound 1013-6 (100.00 mg, 128.38 μmol, 95.51% yield). LCMS (E+) m / z: 779.3 [M+H] + .

[0859] Step 6. Synthesis of Compound 1013

[0860] Compound 1013-6 (100.00 mg, 128.38 μmol) was added to a 20 mL reaction vial and dissolved in anhydrous methanol (3 mL) and water (1 mL). Lithium hydroxide monohydrate (32.35 mg, 770.29 μmol) was then added. The reaction mixture was stirred at room temperature for 2 hours and monitored by LC-MS. After completion of the reaction, the reaction mixture was directly analyzed by MPLC to yield compound 1013 (76.41 mg, 99.89 μmol, 77.81% yield).

[0861] LCMS (E+) m / z: 765.7 [M+H] + .

[0862] 1H NMR (600MHz, Methanol-d4) δ8.34(d,J=8.5Hz,1H),7.74(dd,J=19.0,7.4Hz,2H),7.47(d,J=7.8Hz,3H),7.41(dd,J =7.8,3.8Hz,3H),7.26(d,J=7.8Hz,1H),6.96(dd,J=8.0,2.2Hz,1H),6.80–6.73(m,2H),5.48–5.42(m,1H),4.34(d ,J=4.8Hz,4H),4.25(s,2H),4.16–4.06(m,5H),4.01(t,J=4.5Hz,2H),3.79(dd,J=5.6,3.5Hz,2H),3.72–3.67(m,2 H),3.64(dd,J=5.6,3.3Hz,2H),3.59(d,J=2.7Hz,6H),2.99–2.83(m,4H),2.81(t,J=2.4Hz,1H),2.39–2.36(m,5H).

[0863] Test Example 1 Activity Detection of Compounds Blocking the Binding of Integrin Proteins to Their Ligands

[0864] ELISA assays were used to test the ability of compounds to block the binding of different integrins to their ligands. The following integrins and their ligands were used: αvβ3 and αvβ5 and their ligand rhVitronectin, αvβ6 and αvβ8 and their ligand rhLAP, αvβ1 / α5β1 and their ligand fibonectin, α8β1 and its ligand nephronectin, and α2bβ3 and its ligand fibrinogen. All experimental materials were provided by WuXi AppTec.

[0865] The specific detection method is as follows: prepare the ligand in TBS buffer and transfer 50 μL of the solution to a 96-well plate and incubate at 4°C overnight. Add 150 μL of blocking solution and incubate at 37°C for 1 hour. Integrin protein was prepared using a buffer containing 0.1% BSA. 50 μL of integrin protein was transferred to a 96-well plate, and 1 μL of compound, Ligand SM6.1 (ArrowHead, WO2023070082A2), or DMSO was added. After incubation at room temperature for 2 hours, the plates were washed three times with wash buffer. Antibodies were prepared using a buffer containing 0.1% BSA. αv antibodies were used to detect αvβ3 / αvβ5 / αvβ6 / αvβ1 / αvβ8, α5 antibodies were used to detect α5β1, β1 antibodies were used to detect α8β1, and β3 antibodies were used to detect α2bβ3. 50 μL of the prepared antibodies were added, and the plates were incubated at room temperature for 1 hour. After washing three times with wash buffer, 50 μL of Streptavidin-HRP was added and incubated at room temperature for 20 minutes. 50 μL of TMB substrate was added and incubated at room temperature for 20 minutes. Finally, add 25 μL of stop buffer and read the OD value at 450 nm on a microplate reader. IC was calculated using GraphPad Prism 5 software. 50 value.

[0866] As shown in Tables 2 and 3, compounds 8A and 9B showed superior activity in blocking αvβ6 ligand binding compared to Ligand SM6.1. Compounds 1, 7, 8, 9, 13, 18, 56, 57, and 1008A showed comparable activity to Ligand SM6.1. Compounds 13, 49, 52, 54, 55, 56, 57, 58, and 1013 showed over 50-fold differences in their activity in blocking αvβ6 ligand binding compared to αvβ3 / αvβ5, demonstrating superior αvβ6 selectivity. Ligand SM6.1, compounds 8A, 9B, 13, 1008A, 1009A, and 1013 all blocked αvβ8 ligand binding, but their ability to block αvβ1 / α2bβ3 / α5β1 / α8β1 ligand binding varied.

[0867] Table 2 Activity of compounds in blocking the binding of αvβ3 / αvβ5 / αvβ6 integrin proteins to their ligands

[0868] Table 3 Compounds blocking the binding of αvβ1 / α2bβ3 / αvβ8 / α5β1 / α8β1 integrins to their ligands

[0869] Test Example 2 Compound Blocking Integrin Receptor-Mediated Cell Adhesion Experiment

[0870] To test the ability of compounds to block the binding of cell surface integrins to their ligands, a cell adhesion assay was designed. All experimental materials were provided by WuXi AppTec. Ligands rhVitronectin (recognizing αvβ3 / αvβ5), rhLAP (recognizing αvβ6), and Fibronectin (recognizing α5β1) were prepared in TBS buffer and 50 μL of the solution was transferred to a 96-well plate and coated overnight at 4°C. 150 μL of blocking buffer was added and the plate was blocked at 37°C for 1 hour. SK-MEL-24 (αvβ3), PC-3 (αvβ5), HT-29 (αvβ6), and α5β1-K562 cells were resuspended in buffer and 50 μL was transferred to a 96-well plate. 1 μL of different concentrations of compound, Ligand SM6.1 (ArrowHead, WO2023070082A2), or DMSO was added. SK-MEL-24, HT-29, and α5β1-K562 were incubated at room temperature for 2 hours, and PC-3 was incubated at room temperature for 30 minutes. The liquid in the 96-well plate was removed, and 50 μL of substrate was added and incubated at 37°C for 2 hours. Finally, 90 μL of stop buffer was added, and the OD value was read at 405 nm on a microplate reader. IC was calculated using GraphPad Prism 5 software. 50 value.

[0871] Table 4 Compounds blocking integrin receptor-mediated cell adhesion

[0872] Experimental results:

[0873] All tested compounds effectively blocked cell adhesion mediated by αvβ6 and its ligand. Compounds 13, 8A, 9B, 56, 1008A, 1009A, and 1013 were more active than Ligand SM6.1, while compound 57 was comparable in activity to Ligand SM6.1. Compounds 13, 56, 57, and 1013 exhibited over 50-fold higher blocking activity against αvβ3 / SK-MEL-24, αvβ5 / PC-3, and α5β1 / K562 cells compared to their αvβ6 / HT-29 cell blocking activity, demonstrating αvβ6 specificity.

[0874] Example 25 Preparation of Conjugate 1, Conjugate 2, Conjugate 3, Conjugate 4, Conjugate 5, Conjugate 6, Conjugate 7, Conjugate 8, Conjugate 9, Conjugate 12, Conjugate 13 and Conjugate 14

[0875] Table 5 The present invention relates to conjugates

[0876] 25.1 Oligonucleotide Sequence Synthesis

[0877] The siRNA sequences of the present invention were synthesized using the solid-phase phosphoramidite triester method. The relevant monomers were obtained from Wuhu Huaren Technology Co., Ltd. and Wuhan Tangzhi Pharmaceutical Co., Ltd. Support: 2000A, 500mol synthesis column; coupling time: 4 min; aminolysis conditions: 55°C, concentrated ammonia (25% ammonia solution)

[0878] Sequence synthesis was performed using a 0.2 μmol 48-channel synthesizer. The phosphoramidite monomer was used at a concentration of 0.05 M, and the activator was 0.3 M BTT. The first step, deprotection, involved removing the DMT from the CPG-linked nucleoside with trichloroacetic acid to expose the 5' hydroxyl group for the next coupling step. The second step, activation, involved mixing the monomer with excess tetrazole for activation before loading onto the synthesis column. The third step, coupling, involved a nucleophilic reaction between the phosphoramidite tetrazole and the CPG-linked nucleotide. Following coupling, the tetrazole was removed, thereby extending the oligonucleotide chain. The fourth step, capping, involved esterifying the small amount of 5' hydroxyl group not involved in the coupling reaction with a mixture of acetic anhydride and N-methylimidazole. The fifth step, oxidation, involved sulfhydrylating the trivalent phosphorus to a pentavalent phosphorus using 0.05 M iodine. Repeat steps 1 to 5 until the oligonucleotide chain is extended to the desired length. Cleavage and deprotection: Add concentrated ammonia (25% ammonia in water) to the carrier (CPG) and heat to 55°C for overnight aminolysis.

[0879] After HPLC purification, freeze-drying and quality inspection, the salt was exchanged with sodium acetate alcohol precipitation and desalted using a 3KD ultrafiltration tube. After desalting, the sense chain and antisense chain were quantitatively determined by spectrophotometry, and the sense chain with a C6 amino linker at the end was used for further coupling.

[0880] Table 6 Sequence information

[0881] Synthesis of 25.2TA14 Linker

[0882] Step 1. Synthesis of compound TA14-2

[0883] To a 300 mL solution of compound TA14-1 (25 g, 60.16 mmol), Fmoc-Cl (18.68 g, 72.19 mmol) and sodium bicarbonate (7.58 g, 90.24 mmol) were added. The mixture was stirred at room temperature for 60 minutes until the starting material disappeared as determined by LC-MS. The reaction mixture was directly subjected to MPLC to afford compound TA14-2 (30 g, 47.04 mmol, 78.19% yield). LCMS (E+) m / z: 638.5 [M+H] +

[0884] Step 2. Synthesis of compound TA14-3

[0885] To a dichloromethane solution (300 mL) of compound TA14-2 (30 g, 47.04 mmol) was added trifluoroacetic acid (300 mL) and stirred at room temperature for 10 minutes until the starting material disappeared as determined by LC-MS. The reaction mixture was directly subjected to MPLC to afford compound TA14-3 (28.5 g, 46.40 mmol, 98.65% yield, 95% purity, TF). LCMS (E+) m / z: 470.3 [M+H] +

[0886] Step 3. Synthesis of compound TA14-4

[0887] To a DMF solution (300 mL) of compound TA14-3 (28.5 g, 46.40 mmol, 95% purity, TF) were added 2-[2-(propargyloxy)ethoxy]ethanamine (23.25 g, 162.40 mmol), HATU (61.75 g, 162.40 mmol), and DIPEA (23.99 g, 185.60 mmol, 32.33 mL). The mixture was stirred at room temperature for 2 hours until the starting material disappeared as determined by LC-MS. The reaction mixture was directly subjected to MPLC to afford compound TA14-4 (30 g, 35.50 mmol, 76.52% yield). LCMS (E+) m / z: 845.5 [M+H] +

[0888] Step 4. Synthesis of compound TA14

[0889] To a DMF solution (300 mL) of compound TA14-4 (30 g, 35.50 mmol) was added succinic anhydride (10.13 g, 88.76 mmol) and DIPEA (91.77 g, 710.07 mmol, 123.68 mL). The mixture was stirred at 45°C for 20 hours until the starting material disappeared as determined by LC-MS. The reaction mixture was directly purified by MPLC to afford TA14 (5.1 g, 6.92 mmol, 19.49% yield). LCMS (E+) m / z: 737.7 [M+H] +

[0890] 25.3 Synthesis of Conjugate Intermediate 1

[0891] The oligonucleotide amino compound sense-C6-NH2 was dissolved in sodium borate buffer (250 mM sodium borate, pH = 9.4) to prepare a 4 mM solution (100 μL, 400 nmol). A mixed solution of the TA14 fragment (10 μL, 2000 nmol, 5 equivalents, in 200 mM DMA solution), HATU (5 μL, 2000 nmol, 5 equivalents, in 400 mM DMA solution), and DIPEA (5 μL, 2000 nmol, 5 equivalents, in 400 mM DMA solution) was added to the solution and mixed thoroughly. The reaction solution was then incubated at 25 degrees Celsius for 0.5 hours.

[0892] After the reaction was completed, ethanol precipitation was performed: 10% of the total volume of 5 M sodium chloride solution was added to the solution, followed by the addition of 3 times the total volume of anhydrous ethanol. After vortexing, the reaction mixture was placed on dry ice for 2 hours and then centrifuged at 12,000 rpm for half an hour. The supernatant was discarded and the remaining precipitate was dissolved in deionized water to obtain a solution of conjugate intermediate 1. LC-MS confirmed a reaction conversion rate of 90%.

[0893] 25.4 Synthesis of Conjugate 1, Conjugate 9, and Conjugate 12

[0894] Conjugate intermediate 1 was dissolved in sodium bicarbonate buffer (250 mM sodium bicarbonate, pH = 8.5) to a 1 mM solution (100 μL, 100 nmol). Ligand SM6.1 (30 μL, 3000 nmol, 30 equivalents, 100 mM DMA solution), copper sulfate (10 μL, 1000 nmol, 10 equivalents, 100 mM aqueous solution), and sodium ascorbate (10 μL, 10,000 nmol, 100 equivalents, 1000 mM aqueous solution) were added to the solution and mixed thoroughly. The reaction solution was then incubated at 60°C for 15 minutes.

[0895] After the reaction is complete, ethanol precipitation is performed: 10% of the total volume of 5M sodium chloride solution is added to the solution, followed by the addition of 3 times the total volume of anhydrous ethanol. After oscillation, the reaction is placed on dry ice and frozen for 2 hours. After centrifugation at 12,000 rpm for half an hour, the supernatant is discarded, and the remaining precipitate is dissolved in deionized water. The resulting solution has a reaction conversion rate of 80% as confirmed by LCMS. After reverse phase chromatography purification, the pure product is obtained. After freeze-drying and quality control, the salt is replaced by sodium acetate alcohol precipitation, and desalting is performed using a 3KD ultrafiltration tube. After desalting, the molar amount of the sense chain is quantitatively determined by spectrophotometry. The sense chain and antisense chain are mixed in a 1:1 ratio and annealed to obtain conjugates 1, 9, and 12 duplexes.

[0896] 25.5 Synthesis of Conjugate 2, Conjugate 6, and Conjugate 13

[0897] Conjugate intermediate 1 was dissolved in sodium bicarbonate buffer (250 mM sodium bicarbonate, pH = 8.5) to prepare a 1 mM solution (100 μL, 100 nmol). Compound 8A (30 μL, 3000 nmol, 30 equivalents, 100 mM DMA solution), copper sulfate (10 μL, 1000 nmol, 10 equivalents, 100 mM aqueous solution), and sodium ascorbate (10 μL, 10,000 nmol, 100 equivalents, 1000 mM aqueous solution) were added to the solution and mixed thoroughly. The reaction solution was then placed at 60 degrees Celsius for 15 minutes.

[0898] After the reaction is complete, ethanol precipitation is performed: 10% of the total volume of 5M sodium chloride solution is added to the solution, followed by the addition of 3 times the total volume of anhydrous ethanol. After oscillation, the reaction is placed on dry ice and frozen for 2 hours. After centrifugation at 12,000 rpm for half an hour, the supernatant is discarded, and the remaining precipitate is dissolved in deionized water. The resulting solution has a reaction conversion rate of 75% confirmed by LCMS. After reverse phase chromatography purification, the pure product is obtained. After freeze-drying and quality control, the salt is replaced by sodium acetate alcohol precipitation, and desalting is performed using a 3KD ultrafiltration tube. After desalting, the molar amount of the sense chain is quantitatively determined by spectrophotometry. The sense chain and antisense chain are mixed in a 1:1 ratio and annealed to obtain the target duplexes of conjugates 2, conjugate 6, and conjugate 13.

[0899] 25.6 Synthesis of Conjugate 3 and Conjugate 7

[0900] Note: * indicates that the stereo configuration of the three carbon atoms has not yet been determined. They may all be R configuration or S configuration.

[0901] Conjugate intermediate 1 was dissolved in sodium bicarbonate buffer (250 mM sodium bicarbonate, pH = 8.5) to a 1 mM solution (100 μL, 100 nmol). Compound 9B (30 μL, 3000 nmol, 30 equivalents, 100 mM DMA solution), copper sulfate (10 μL, 1000 nmol, 10 equivalents, 100 mM aqueous solution), and sodium ascorbate (10 μL, 10,000 nmol, 100 equivalents, 1000 mM aqueous solution) were added to the solution and mixed thoroughly. The reaction solution was then incubated at 60°C for 15 minutes.

[0902] After the reaction is complete, ethanol precipitation is performed: 10% of the total volume of 5M sodium chloride solution is added to the solution, followed by the addition of 3 times the total volume of anhydrous ethanol. After vortexing, the reaction is placed on dry ice and frozen for 2 hours. The mixture is then centrifuged at 12,000 rpm for half an hour. The supernatant is discarded and the remaining precipitate is dissolved in deionized water. The resulting solution has a reaction conversion rate of 75% as confirmed by LCMS. After reverse-phase chromatography purification, the pure product is obtained. After freeze-drying and quality control, the salt is exchanged with sodium acetate alcohol precipitation and desalted using a 3KD ultrafiltration tube. After desalting, the molar amount of the sense chain is quantitatively determined by spectrophotometry. The sense and antisense chains are mixed in a 1:1 ratio and annealed to obtain the target duplexes of conjugates 3 and 7.

[0903] 25.7 Synthesis of Conjugates 4, 8, and 14

[0904] Conjugate intermediate 1 was dissolved in sodium bicarbonate buffer (250 mM sodium bicarbonate, pH = 8.5) to prepare a 1 mM solution (100 μL, 100 nmol). Compound 13 (30 μL, 3000 nmol, 30 equivalents, 100 mM DMA solution), copper sulfate (10 μL, 1000 nmol, 10 equivalents, 100 mM aqueous solution), and sodium ascorbate (10 μL, 10,000 nmol, 100 equivalents, 1000 mM aqueous solution) were added to the solution and mixed well. The reaction solution was then placed at 60 degrees Celsius for 15 minutes.

[0905] After the reaction is complete, ethanol precipitation is performed: 10% of the total volume of 5M sodium chloride solution is added to the solution, followed by the addition of 3 times the total volume of anhydrous ethanol. After oscillation, the reaction is placed on dry ice and frozen for 2 hours. After centrifugation at 12,000 rpm for half an hour, the supernatant is discarded, and the remaining precipitate is dissolved in deionized water. The resulting solution has a reaction conversion rate of 80% as confirmed by LCMS. After reverse phase chromatography purification, the pure product is obtained. After freeze-drying and quality control, the salt is replaced by sodium acetate alcohol precipitation, and desalting is performed using a 3KD ultrafiltration tube. After desalting, the molar amount of the sense chain is quantitatively determined by spectrophotometry. The sense chain and antisense chain are mixed in a 1:1 ratio and annealed to obtain the target duplexes of conjugates 4, 8, and 14.

[0906] 25.8. Synthesis of Conjugate 5

[0907] 25.8.1 Synthesis of Linker L1001

[0908] Step 1. Synthesis of compound L1001-2

[0909] To a DMF solution (30 mL) of compound L1001-1 (3 g, 6.39 mmol) was added 2-[2-(2-azidoethoxy)ethoxy]ethanamine' (3.90 g, 22.37 mmol), HATU (8.50 g, 22.37 mmol), and DIPEA (3.30 g, 25.56 mmol, 4.45 mL). The mixture was stirred at room temperature for 2 hours until the starting material disappeared as determined by LC-MS. The reaction mixture was directly subjected to MPLC to afford compound L1001-2 (4.1 g, 4.37 mmol, 68.40% yield). LCMS (E+) m / z: 845.5 [M+H] +

[0910] Step 2. Synthesis of compound L1001

[0911] To a DMF solution (33 mL) of compound L1001-2 (4.1 g, 4.37 mmol) were added glutaric anhydride (1.50 g, 13.11 mmol) and DIPEA (11.30 g, 87.42 mmol, 15.23 mL). The mixture was stirred at 45°C for 20 hours until the starting material disappeared as determined by LC-MS. The reaction mixture was directly purified by MPLC to afford L1001 (1.05 g, 1.27 mmol, 28.95% yield). LCMS (E+) m / z: 830.7 [M+H] +

[0912] 25.8.2 Synthesis of Conjugate Intermediate 2

[0913] The oligonucleotide amino compound sense strand -C6-NH2 was dissolved in sodium borate buffer (250 mM sodium borate, pH = 9.4) to prepare a 4 mM solution (100 μL, 400 nmol). A mixed solution of linker L1001 (10 μL, 2000 nmol, 5 equivalents, in 200 mM DMA solution), HATU (5 μL, 2000 nmol, 5 equivalents, in 400 mM DMA solution), and DIPEA (5 μL, 2000 nmol, 5 equivalents, in 400 mM DMA solution) was added to the solution and mixed thoroughly. The reaction solution was then incubated at 25°C for 0.5 h.

[0914] After the reaction was completed, ethanol precipitation was performed: 10% of the total volume of 5 M sodium chloride solution was added to the solution, followed by the addition of 3 times the total volume of anhydrous ethanol. After vortexing, the reaction mixture was placed on dry ice for 2 hours and then centrifuged at 12,000 rpm for half an hour. The supernatant was discarded and the remaining precipitate was dissolved in deionized water to obtain a solution of conjugate intermediate 2. LC-MS confirmed that the reaction conversion rate was 80%.

[0915] 25.8.3 Synthesis of Conjugate 5

[0916] Conjugate intermediate 2 was dissolved in sodium bicarbonate buffer (250 mM sodium bicarbonate, pH = 8.5) to a 1 mM solution (100 μL, 100 nmol). Compound 1008A (30 μL, 3000 nmol, 30 equivalents, 100 mM DMA solution), copper sulfate (10 μL, 1000 nmol, 10 equivalents, 100 mM aqueous solution), and sodium ascorbate (10 μL, 10,000 nmol, 100 equivalents, 1000 mM aqueous solution) were added to the solution and mixed thoroughly. The reaction solution was then incubated at 60°C for 15 minutes.

[0917] After the reaction is complete, ethanol precipitation is performed: 10% of the total volume of 5M sodium chloride solution is added to the solution, followed by 3 times the total volume of anhydrous ethanol. After vortexing, the reaction mixture is placed on dry ice and chilled for 2 hours. The mixture is then centrifuged at 12,000 rpm for half an hour. The supernatant is discarded and the remaining precipitate is dissolved in deionized water. The resulting solution has a reaction conversion rate of 70% as confirmed by LCMS. Purification by reverse-phase chromatography yields the pure product. After lyophilization and quality control, the salt is exchanged with sodium acetate alcohol precipitation and desalted using a 3KD ultrafiltration tube. After desalting, the molar weight of the sense strand is quantified by spectrophotometry. The sense and antisense strands are mixed in a 1:1 ratio and annealed to obtain the target duplex of Conjugate 5.

[0918] Test Example 3: Knockdown activity detection of conjugates 1-4 in humanized MMP7 mice

[0919] In vivo administration: Humanized MMP7 homozygous female mice (purchased from Biocytogen), the transgenic mice contain the full-length human MMP7 gene (CDS+UTR). In the in vivo experiment, each group of 4 mice, female, 6-8 weeks old. After one week of breeding and adaptation in an SPF environment, the conjugate was administered. The administration method was: nebulized needle airway administration, the dosage was 1 mg / kg, the conjugate was dissolved in normal saline, and the administration volume was 50 μL. The normal saline group did not contain the conjugate, but only contained 50 μL of normal saline. 14 days after administration, the mice were anesthetized and killed, and the alveolar lavage fluid was first obtained with pre-cooled PBS to detect the expression level of MMP7 protein. After perfusion, lung tissue was obtained to detect the mRNA expression level of MMP7.

[0920] RNA expression detection: After removing the lung tissue, add tissue protection solution and place it at 4°C overnight, then place it in a -80°C refrigerator for long-term storage. Discard the tissue protection solution, add the prepared tissue lysis solution according to the instructions of TaKaRa Mini BEST Universal RNAExtraction Kit (Takara: 9767), and add magnetic beads to homogenize on a tissue homogenizer. After the homogenization is completed, perform the subsequent RNA extraction steps according to the instructions. Use the Takara reverse transcription kit (RR036A) to reverse transcribe the extracted RNA, and use the reverse transcribed cDNA to Green kit (ACR, AG11718) was used to amplify the target gene expression level. GAPDH was used as the internal reference and the ΔΔCt method was used in ABIQuantStudio TM Real-time fluorescence PCR was performed in a 6-well real-time fluorescence PCR system. Two replicate wells were prepared for each mouse lung tissue sample.

[0921] Protein Expression Assay: After obtaining bronchoalveolar lavage fluid, store it in a -80°C freezer for long-term storage. Thaw on ice and centrifuge the supernatant for protein quantification using Pierce BCA Protein Assay Kits (Thermo-23225). MMP7 protein expression was assayed using an MMP7 assay kit (Wuhan Cloud-Clone Technology Co., Ltd., SEA102Hu).

[0922] Table 7 Expression levels of MMP7 mRNA and protein in MMP7 mice after aerosolized airway administration

[0923] The results are shown in Table 7. The conjugates coupled with siMMP7 small nucleic acid were able to downregulate the expression of MMP7 mRNA in vivo. The ability of conjugate 4 (coupled with compound 13) to downregulate MMP7 mRNA was comparable to that of conjugate 1 (coupled with Ligand SM6.1); while conjugate 2 (coupled with 8A) and conjugate 3 (coupled with 9B) were significantly better than conjugate 1 in downregulating MMP7 mRNA and protein.

[0924] Test Example 4: Activity detection of conjugate 6-9 in wild-type C57BL / 6N mice

[0925] In vivo administration: C57BL / 6N mice were purchased from Weitonglihua. In the in vivo experiment, each group consisted of 3-5 male mice, 6-8 weeks old. The administration method and dosage were the same as those in Test Example 3. The mice were anesthetized and killed after 7 and 14 days, respectively. After the lung tissues were removed from the body, they were quickly frozen in liquid nitrogen and placed in a -80°C refrigerator for long-term storage. According to the instructions of TaKaRa MiniBEST Universal RNA Extraction Kit (Takara: 9767), the prepared tissue lysis buffer was added, and magnetic beads were added for homogenization on a tissue homogenizer. After homogenization, the subsequent RNA extraction steps were carried out according to the instructions. Use One Step PrimeScript TM Gene expression was detected using an RT-PCR Kit (Takara: RR064A). Real-time PCR was performed using the ΔΔCt method in an ABIQS5 real-time PCR system. Three replicate wells were performed for each mouse lung tissue sample.

[0926] Table 8 Relative expression levels of RAGE mRNA in C57BL / 6N mice at different time points after aerosol airway administration

[0927] The results are shown in Table 8. The conjugates coupled to the siRAGE small nucleic acid were able to downregulate the expression of RAGE mRNA levels in vivo. At the 7th day time point, conjugate 6 (coupled with compound 8A) and conjugate 8 (coupled with compound 13) were more effective at downregulating RAGE mRNA than conjugate 9 (coupled with Ligand SM6.1). At the 14th day time point, conjugate 6 and conjugate 7 (coupled with compound 9B) were both more effective at downregulating RAGE mRNA than conjugate 9.

[0928] Test Example 5: Activity detection of conjugate 5 and conjugate 12 in wild-type C57BL / 6N mice

[0929] In vivo administration: C57BL / 6N mice were purchased from Weitonglihua. In the in vivo experiment, each group consisted of 3-5 male mice, 6-8 weeks old. The administration method was the same as in Test Example 3, with a dose of 10 mg / kg. After 10 days, the mice were anesthetized and killed. After the lung tissue was removed from the body, it was quickly frozen in liquid nitrogen and placed in a -80°C refrigerator for long-term storage. According to the instructions of the TaKaRa MiniBEST Universal RNA Extraction Kit (Takara: 9767), the prepared tissue lysis buffer was added, and magnetic beads were added for homogenization on a tissue homogenizer. After homogenization, the subsequent RNA extraction steps were performed according to the instructions. Use One Step PrimeScript TMGene expression was detected using an RT-PCR Kit (Takara: RR064A). Real-time PCR was performed using the ΔΔCt method in an ABIQS5 real-time PCR system. Three replicate wells were performed for each mouse lung tissue sample.

[0930] Table 9 Relative expression levels of SOD1 mRNA in C57BL / 6N mice at different time points after aerosol airway administration

[0931] The results are shown in Table 9. All conjugates coupled with siSOD1 small nucleic acid were able to downregulate SOD1 mRNA expression in vivo. At the 10th day time point, conjugate 5 (coupled with compound 1008A) was significantly more effective in downregulating SOD1 mRNA than conjugate 12 (coupled with Ligand SM6.1).

Claims

1. A compound represented by formula I or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, in: X is selected from direct bond, Preferably, X is selected from a direct bond or More preferably, X is a direct bond; Y is selected from NH or CH2; n is 0, 1 or 2; m is 0 or 1; Preferably, m is 0; R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl; R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 alkyl; Ring B is selected from 3-10 membered cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aromatic ring, 5-10 membered aromatic heterocycle; R 4 Selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 alkoxy, optionally substituted amino; R 5a 、R 5b are independently selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 alkoxy, optionally substituted amino; Preferably, R 5a 、R 5b are each independently selected from hydrogen, halogen, -C 1～6 alkyl; More preferably, R 5a 、R 5b are each independently selected from hydrogen, -C 1～6 alkyl; Most preferably, R 5a 、R 5b are each independently selected from hydrogen, methyl; Or, R 4 and R 5a Directly connected to form a C containing 1-3 heteroatoms 3～6 Alkylene chains (preferably forming )； Ring A is selected from 5-10 membered aromatic heterocycles and 3-10 membered heterocycloalkyls, wherein the aromatic heterocycles and heterocycloalkyls are optionally replaced by one, two or three R A replace; Each R A are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl); Q is selected from -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C(O)NR Q4 -、-C 0～4 Alkylene-(3-10 membered cycloalkyl)-C 0～4 Alkylene-NR Q4 -、-C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C 0～4 Alkylene, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C(O)NR Q4 -、-C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-NR Q4 -、-C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-O-, -C 0～4 Alkylene-CR Q1 R Q2 -NR Q4 -、-C 0～4 Alkylene-(5-12 membered spiroheterocyclic)-, -C 0～4 Alkylene-(5-12 membered fused heterocyclic ring)-, -C 0～4 Alkylene-(5-10 membered aromatic heterocycle)-, C 0～4 Alkylene-(5-10 membered aromatic ring)-NR Q4 -、 The cycloalkyl, heterocycloalkyl, spiro heterocycle, fused heterocycle, aromatic heterocycle, aromatic ring are optionally substituted by one, two or three R Q3 replace; Each R Q3 are independently selected from hydrogen, halogen, cyano, =O, nitro, -OH, -C 1～6 Alkyl, halogen-substituted -C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl); Each R Q4 are independently selected from hydrogen, -C 1～6 Alkyl, halogen-substituted -C 1～6 alkyl; Preferably, each R Q4 are independently selected from hydrogen, -C 1～6 alkyl; More preferably, each R Q4 are independently selected from hydrogen, -C 1～3 alkyl; Most preferably, each R Q4 are independently selected from hydrogen, methyl; n2 is an integer from 0 to 6; The R Q1 、R Q2 The atoms directly connected to it form a 3-10 membered heterocyclic ring; The L1 is selected from Preferably, the L1 is selected from When X is selected from a direct bond, Y is selected from NH, and Q is selected from R 1 、R 2 When selected from hydrogen, the L1 is not selected from 2. The compound according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Ring A is selected from 5-membered aromatic heterocycle, 6-membered aromatic heterocycle, 9-membered aromatic heterocycle, 5-membered heterocycloalkyl, 6-membered heterocycloalkyl, 9-membered heterocycloalkyl, 10-membered heterocycloalkyl; the aromatic heterocycle and heterocycloalkyl are optionally replaced by one, two or three R A replace; Preferably, ring A is selected from and / or, Each R A are independently selected from hydrogen, =O, -C 1～3 Alkyl, -O(C 1～6 Alkyl), halogen-substituted C 1～3 Alkyl, halogen, -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl); Preferably, each R A Each is independently selected from hydrogen, =0, methyl, isopropyl, methoxy, trifluoromethyl, fluorine, -NH2, and -NH(CH3).

3. The compound according to any one of claims 1 to 2, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Q is selected from -(7-membered spiroheterocycle)-, -(8-membered spiroheterocycle)-, -(9-membered spiroheterocycle)-, -(10-membered spiroheterocycle)-, -(8-membered fused heterocycle)-, -C3 alkylene-(5-membered aromatic heterocycle)-, -C1 alkylene-(3-membered cycloalkyl)-C(O)NH-, -(4-membered cycloalkyl)-C1 alkylene-NR Q4 -, -(5-membered cycloalkyl)-C(O)NH-, -(6-membered cycloalkyl)-C(O)NH-, -C2 alkylene-(5-membered cycloalkyl)-NH-, -C1 alkylene-(5-membered cycloalkyl)-NH-, -C1 alkylene-(5-membered cycloalkyl)-C1 alkylene-NH-, -C1 alkylene-(5-membered heterocycloalkyl)-, -C2 alkylene-(4-membered heterocycloalkyl)-, -C2 alkylene-(5-membered heterocycloalkyl)-, -C3 alkylene -(5-membered heterocycloalkyl)-, -C1 alkylene-(6-membered heterocycloalkyl)-, -C1 alkylene-(5-membered heterocycloalkyl)-NH-, -C1 alkylene-(5-membered heterocycloalkyl)-O-, -(5-membered heterocycloalkyl)-C(O)NH-, -(6-membered heterocycloalkyl)-C(O)NH-, -C1 alkylene-(5-membered heterocycloalkyl)-C1 alkylene, -C1 alkylene-(6-membered heterocycloalkyl)-C1 alkylene, -C3 alkylene-CR Q1 R Q2 -NH-, -C1 alkylene-phenyl ring-NR Q4 -; the cycloalkyl, heterocycloalkyl, spiro heterocycle, fused heterocycle, aromatic heterocycle, benzene ring is optionally replaced by one, two or three R Q3 replace; Preferably, Q is selected from and / or, Each R Q3 are independently selected from hydrogen, =O, -C 1～3 Alkyl, halogen-substituted -C 1～3 alkyl; Preferably, each R Q3 are independently selected from hydrogen, =O, -C 1～3 alkyl; More preferably, each R Q3 are independently selected from hydrogen, =O, methyl; and / or, The R Q1 、R Q2 The atoms directly connected thereto form a 4-membered heterocyclic ring (preferably forming oxetane).

4. The compound according to any one of claims 1 to 3, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Ring B is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, phenyl, naphthyl, pyridyl, pyrimidinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl, thienyl, benzothienyl, thiazolyl, benzothiazolyl, furyl, oxazolyl, isoxazolyl, benzofuranyl, indolyl, indazolyl, benzimidazolyl, oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, tetrazolyl, quinolyl, isoquinolyl, quinoxalinyl, tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, pyrrolidinyl, dioxanyl, or dioxolanyl; Preferably, Ring B is naphthyl; More preferably, Ring B is 5. The compound according to any one of claims 1 to 4, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: R 1 、R 2 are independently selected from hydrogen and fluorine; and / or, R 3 Selected from hydrogen, -C 1～3 alkyl; Preferably, R 3 For hydrogen.

6. The compound according to any one of claims 1 to 5, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: The compound has the structure shown in formula I-1, in: X is selected from direct bond, Preferably, X is selected from a direct bond or More preferably, X is a direct bond; Y is selected from NH or CH2; Preferably, Y is NH; n is 0, 1 or 2; m is 0 or 1; R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl; Preferably, R 1 、R 2 are independently selected from hydrogen and halogen; More preferably, R 1 、R 2 are independently selected from hydrogen and fluorine; Most preferably, R 1 、R 2 is hydrogen; R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 alkyl; Preferably, R 3 Selected from hydrogen, -C 1～3 alkyl; More preferably, R 3 is hydrogen; R 5a 、R 5b are independently selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 alkoxy, optionally substituted amino; Preferably, R 5a 、R 5b are each independently selected from hydrogen, halogen, -C 1～6 alkyl; More preferably, R 5a 、R 5b are each independently selected from hydrogen, -C 1～6 alkyl; Most preferably, R 5a 、R 5b are each independently selected from hydrogen, methyl; R A1 、R A2 、R A3 are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl); Preferably, R A1 、R A2 、R A3 are independently selected from hydrogen, =O, -C 1～3 Alkyl, -O(C 1～6 Alkyl), halogen-substituted C 1～3 Alkyl, halogen, -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl); More preferably, R A1 、R A2 、R A3 are independently selected from hydrogen, -C 1～3 Alkyl, -O(C 1～6 Alkyl), halogen-substituted C 1～3 Alkyl, halogen, -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl); More preferably, R A1 、R A2 、R A3 Each of the following groups is independently selected from hydrogen, methyl, isopropyl, methoxy, trifluoromethyl, fluorine, -NH2, -NH(CH3); Most preferably, R A1 、R A2 、R A3 are independently selected from hydrogen, methyl, isopropyl, methoxy (preferably, R A1 、R A2 is hydrogen, R A3 is selected from hydrogen, methyl, isopropyl, methoxy); Q is selected from -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered cycloalkyl)-C 0～4 Alkylene-NR Q4 -、-C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C 0～4 Alkylene, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-C(O)NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-NH-, -C 0～4 Alkylene-(3-10 membered heterocycloalkyl)-O-, -C 0～4 Alkylene-(5-12 membered spiroheterocyclic)-, -C 0～4 Alkylene-(5-12 membered fused heterocyclic ring)-, -C 0～4 Alkylene-(5-10 membered aromatic heterocycle)-, C 0～4 Alkylene-(5-10 membered aromatic ring)-NR Q4 -; the cycloalkyl, heterocycloalkyl, spiro heterocycle, fused heterocycle, aromatic heterocycle, aromatic ring are optionally substituted by one, two or three R Q3 replace; Preferably, Q is selected from -(7-membered spiroheterocycle)-, -(8-membered spiroheterocycle)-, -(9-membered spiroheterocycle)-, -(10-membered spiroheterocycle)-, -(8-membered fused heterocycle)-, -C3 alkylene-(5-membered aromatic heterocycle)-, -C1 alkylene-(3-membered cycloalkyl)-C(O)NH-, -(4-membered cycloalkyl)-C1 alkylene-NR Q4 -、-(5-membered cycloalkyl)-C(O)NH-、-(6-membered cycloalkyl)-C(O)NH-、-C1 alkylene-(5-membered cycloalkyl)-NH-、-C1 alkylene-(5-membered cycloalkyl)-C1 alkylene-NH-、-C1 alkylene-(5-membered heterocycloalkyl)-、-C2 alkylene-(4-membered heterocycloalkyl)-、-C2 alkylene-(5-membered heterocycloalkyl)-、-C3 alkylene-(5-membered heterocycloalkyl)-、-C1 alkylene-(6-membered heterocycloalkyl)-、-C1 alkylene-(5-membered heterocycloalkyl)-NH-、-C1 alkylene-(5-membered heterocycloalkyl)-O-、-(6-membered heterocycloalkyl)-C(O)NH-、-C1 alkylene-(6-membered heterocycloalkyl)-C1 alkylene、-C1 alkylene-phenyl ring-NR Q4 -; the cycloalkyl, heterocycloalkyl, spiro heterocycle, fused heterocycle, aromatic heterocycle, benzene ring is optionally replaced by one, two or three R Q3 replace; Each R Q3 are independently selected from hydrogen, halogen, cyano, =O, nitro, -OH, -C 1～6 Alkyl, halogen-substituted -C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl); Preferably, each R Q3 are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted -C 1～6 alkyl; More preferably, each R Q3 are independently selected from hydrogen, =O, -C 1～3 Alkyl, halogen-substituted -C 1～3 alkyl; Further preferably, each R Q3 are independently selected from hydrogen, =O, -C 1～3 alkyl; Most preferably, each R Q3 are independently selected from hydrogen, =O, methyl; Each R Q4 are independently selected from hydrogen, -C 1～6 Alkyl, halogen-substituted -C 1～6 alkyl; Preferably, each R Q4 are independently selected from hydrogen, -C 1～6 alkyl; More preferably, each R Q4 Each independently selected from -C 1～3 alkyl; Most preferably, R Q4 is methyl; More preferably, Q is selected from 7. The compound according to any one of claims 1 to 5, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: The compound has the structure shown in formula I-2, n is 0, 1 or 2, preferably 1; n2 is 0, 1, 2, 3, 4, 5 or 6, preferably 0, 1, 2, more preferably 1; R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl; Preferably, R 1 、R 2 are independently selected from hydrogen and halogen; More preferably, R 1 、R 2 are independently selected from hydrogen and fluorine; Most preferably, R 1 、R 2 is hydrogen; R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 alkyl; Preferably, R 3 Selected from hydrogen, -C 1～3 alkyl; More preferably, R 3 is hydrogen; Ring A is selected from 5-10 membered aromatic heterocycles and 3-10 membered heterocycloalkyls, wherein the aromatic heterocycles and heterocycloalkyls are optionally replaced by one, two or three R A replace; Preferably, the A ring is selected from a 6-membered aromatic heterocycle, a 9-membered aromatic heterocycle, a 9-membered heterocycloalkyl, or a 10-membered heterocycloalkyl; the aromatic heterocycle or heterocycloalkyl is optionally replaced by one, two, or three R A replace; Each R A are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl); Preferably, each R A are independently selected from =O, -C 1～3 Alkyl, halogen-substituted C 1～3 Alkyl, halogen, -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl); More preferably, each R A Each independently selected from =O, methyl, trifluoromethyl, fluorine, -NH2, -NH(CH3); Preferably, ring A is selected from 8. The compound according to any one of claims 1 to 5, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: The compound has the structure shown in formula I-3, in: n is 0, 1 or 2, preferably 1; n2 is 0, 1, 2, 3, 4, 5 or 6, preferably 1, 2, 3; R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl; Preferably, R 1 、R 2 are independently selected from hydrogen and halogen; More preferably, R 1 、R 2 are independently selected from hydrogen and fluorine; R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 alkyl; Preferably, R 3 Selected from hydrogen, -C 1～3 alkyl; More preferably, R 3 is hydrogen; R A1 、R A2 、R A3 are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl); Preferably, R A1 、R A2 、R A3 are independently selected from hydrogen, =O, -C 1～3 Alkyl, halogen-substituted C 1～3 Alkyl, halogen, -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl); More preferably, R A1 、R A2 、R A3 are independently selected from hydrogen, -C 1～3 Alkyl, halogen-substituted C 1～3 Alkyl, halogen, -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl); More preferably, R A1 、R A2 、R A3 Each independently selected from hydrogen, methyl, trifluoromethyl, fluorine, -NH2, -NH(CH3); Most preferably, R A1 、R A2 、R A3 are independently selected from hydrogen, methyl (preferably, R A1 、R A2 is hydrogen, R A3 is methyl); L is selected from When R 1 、R 2 When it is hydrogen, L is not 9. The compound according to any one of claims 1 to 5, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: The compound has the structure shown in formula I-4, in: R 1 、R 2 are independently selected from hydrogen, halogen, -C 1～6 Alkyl, halogen-substituted C 1～6 alkyl; Preferably, R 1 、R 2 are independently selected from hydrogen and halogen; More preferably, R 1 、R 2 are independently selected from hydrogen and fluorine; Most preferably, R 1 、R 2 is hydrogen; R 3 Selected from hydrogen, -C 1～6 Alkyl, optionally substituted C 1～6 alkyl; Preferably, R 3 Selected from hydrogen, -C 1～3 alkyl; More preferably, R 3 is hydrogen; R 5a 、R 5b are each independently selected from hydrogen, halogen, -C 1～6 Alkyl, optionally substituted C 1～6 Alkyl, -O(C 1～6 alkyl), optionally substituted C 1～6 alkoxy, optionally substituted amino; Preferably, R 5a 、R 5b are each independently selected from hydrogen, halogen, -C 1～6 alkyl; More preferably, R 5a 、R 5b are each independently selected from hydrogen, -C 1～6 alkyl; Most preferably, R 5a 、R 5b are each independently selected from hydrogen, methyl; Ring A is selected from 5-10 membered aromatic heterocycles and 3-10 membered heterocycloalkyls, wherein the aromatic heterocycles and heterocycloalkyls are optionally replaced by one, two or three (preferably one) R A replace; Preferably, the A ring is selected from a 6-membered aromatic heterocycle, a 9-membered heterocycloalkyl, or a 10-membered heterocycloalkyl; the aromatic heterocycle or heterocycloalkyl is optionally replaced by one, two, or three (preferably one) R A replace; More preferably, ring A is a benzene ring; the benzene ring is optionally replaced by one, two or three (preferably one) R A replace; Each R A are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted C 1～6 Alkyl, halogen, cyano, nitro, -OH, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 Alkyl), -C 0～2 Alkylene-(3-10 membered cycloalkyl), -C 0～2 Alkylene-(3- to 10-membered heterocycloalkyl); Preferably, each R A are independently selected from hydrogen, -C 1～3 Alkyl, halogen-substituted C 1～3 Alkyl, -OH, -O(C 1～6 alkyl); More preferably, each R A are independently selected from hydrogen, -C 1～3 Alkyl, -O(C 1～6 alkyl); Most preferably, each R A are independently selected from hydrogen, methyl, isopropyl, and methoxy; More preferably, ring A is selected from Most preferably, Ring A is selected from L0 is selected from n2 is 0, 1, 2, 3, 4, 5 or 6, preferably 1, 2, 3, more preferably 1; Q is selected from -C 0～4 Alkylene-(5-12 membered spiroheterocyclic)-, -C 0～4 Alkylene-(5-12 membered fused heterocycle)-; the spiro heterocycle and fused heterocycle are optionally replaced by one, two or three R Q3 replace; Preferably, Q is selected from -(7-membered spiro heterocycle)-, -(8-membered spiro heterocycle)-, -(9-membered spiro heterocycle)-, -(8-membered fused heterocycle)-; the spiro heterocycle and fused heterocycle are optionally replaced by one, two or three R Q3 replace; Or preferably, Q is selected from -C 0～4 Alkylene-(5-12 membered spiroheterocycle)-; the spiroheterocycle is optionally replaced by one, two or three R Q3 replace; Or preferably, Q is selected from -(7-membered spiro heterocycle)-, -(8-membered spiro heterocycle)-, -(9-membered spiro heterocycle)-; the spiro heterocycle is optionally replaced by one, two or three R Q3 replace; Each R Q3 are independently selected from hydrogen, halogen, cyano, =O, nitro, -OH, -C 1～6 Alkyl, halogen-substituted -C 1～6 Alkyl, -O(C 1～6 Alkyl), -O(halogen substituted C 1～6 alkyl), -NH2, -NH(C 1～6 Alkyl), -N(C 1～6 Alkyl)(C 1～6 alkyl); Preferably, each R Q3 are independently selected from hydrogen, =O, -C 1～6 Alkyl, halogen-substituted -C 1～6 alkyl; More preferably, each R Q3 are independently selected from hydrogen, =O, -C 1～3 Alkyl, halogen-substituted -C 1～3 alkyl; Further preferably, each R Q3 are independently selected from hydrogen, =O, -C 1～3 alkyl; More preferably, each R Q3 are independently selected from hydrogen, =O; Most preferably, each R Q3 is hydrogen; More preferably, Q is selected from More preferably, Q is selected from Most preferably, Q is selected from Preferably, L0 is selected from More preferably, L0 is selected from Most preferably, L0 is selected from 10. The compound according to any one of claims 1 to 9, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: The compound is selected from: Further selected from:

11. An αvβ6 integrin ligand comprising a targeting moiety, wherein the targeting moiety comprises the compound according to any one of claims 1 to 10, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a group formed by removing one or more (preferably one) hydrogen atoms from the compound, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

12. The αvβ6 integrin ligand according to claim 11, wherein The compound has the structure shown in Formula I, X, Y, n, m, R 1 、R 2 、R 3 、R 4 、R 5a 、R 5b , Ring A, Ring B, Q, and L1 are each independently as defined in any one of claims 1 to 5; Wherein, when the targeting portion comprises a group formed by removing one or more (preferably one) hydrogen atoms from the compound or its stereoisomer or its pharmaceutically acceptable salt, in R 3 、R 4 or removing one or more (preferably one) hydrogen atoms from the B ring (preferably from the B ring); Preferably, Ring B is When the targeting moiety comprises a group represented by Formula Ia, X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b , Ring A, Q, and L1 are each independently as defined in any one of claims 1 to 5, 13. The αvβ6 integrin ligand according to any one of claims 11 to 12, wherein The compound has the structure shown in formula I-1, X, Y, n, m, R 1 、R 2 、R 3 、R A1 、R A2 、R A3 、R 5a 、R 5b , Q are each independently as defined in claim 6, Wherein, when the targeting portion comprises a group formed by removing one or more (preferably one) hydrogen atoms from the compound or its stereoisomer or its pharmaceutically acceptable salt, in R 3 or Above (preferably ) removing one or more (preferably one) hydrogen atoms; Preferably, the targeting moiety comprises a group represented by formula I-1a, wherein X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b 、R A1 、R A2 、R A3 , Q are each independently as defined in claim 6, 14. The αvβ6 integrin ligand according to any one of claims 11 to 12, wherein The compound has the structure shown in formula I-2, n、n2、R 1 、R 2 、R 3 , Ring A are each independently as defined in claim 7, Wherein, when the targeting portion comprises a group formed by removing one or more (preferably one) hydrogen atoms from the compound or its stereoisomer or its pharmaceutically acceptable salt, in R 3 or Above (preferably ) removing one or more (preferably one) hydrogen atoms; Preferably, the targeting moiety comprises a group represented by formula I-2a, wherein n, n2, R 1 、R 2 、R 3 , Ring A are each independently as defined in claim 7, 15. The αvβ6 integrin ligand according to any one of claims 11 to 12, wherein The compound has the structure shown in formula I-3, n、n2、R 1 、R 2 、R 3 、R A1 、R A2 、R A3 , L are each independently as defined in claim 8, Wherein, when the targeting portion comprises a group formed by removing one or more (preferably one) hydrogen atoms from the compound or its stereoisomer or its pharmaceutically acceptable salt, in R 3 or Above (preferably ) removing one or more (preferably one) hydrogen atoms; Preferably, the targeting moiety comprises a group shown in formula I-3a, wherein n, n2, R 1 、R 2 、R 3 、R A1 、R A2 、R A3 , L are each independently as defined in claim 8, 16. The αvβ6 integrin ligand according to any one of claims 11 to 12, wherein The compound has the structure shown in formula I-4, R 1 、R 2 、R 3 、R 5a 、R 5b , L0, and A ring are each independently as defined in claim 9, Wherein, when the targeting portion comprises a group formed by removing one or more (preferably one) hydrogen atoms from the compound or its stereoisomer or its pharmaceutically acceptable salt, in R 3 or Above (preferably ) removing one or more (preferably one) hydrogen atoms; Preferably, the targeting moiety comprises a group represented by formula I-4a, R 1 、R 2 、R 3 、R 5a 、R 5b , L0, and A ring are each independently as defined in claim 9, 17. The αvβ6 integrin ligand according to any one of claims 11 to 16, wherein The targeting moiety comprises the compound of claim 10 or comprises a group selected from the group consisting of:

18. The αvβ6 integrin ligand according to any one of claims 11 to 17, wherein The αvβ6 integrin ligand further comprises a linker moiety covalently attached to the targeting moiety; Preferably, the linker moiety comprises a PEG linker; Preferably, the linker portion is n1 is selected from integers of 2 to 20, preferably from integers of 2 to 10.

19. The αvβ6 integrin ligand according to claim 18, wherein The targeting portion comprises a group formed by removing a hydrogen atom from the compound shown in Formula I or its stereoisomer or a pharmaceutically acceptable salt thereof, and the linker portion is The linker moiety is covalently linked to the targeting moiety, X, Y, n, m, R 1 、R 2 、R 3 、R 4 、R 5a 、R 5b , Ring A, Ring B, Q, and L1 are each independently as defined in any one of claims 1 to 5, and n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10; Wherein, the targeting portion comprises a compound of formula I or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in R 3 、R 4 or a group formed by removing a hydrogen atom from the B ring (preferably from the B ring); Preferably, Ring B is When the αvβ6 integrin ligand comprises a group represented by formula Ib, X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b , Ring A, Q, L1 are each independently as defined in any one of claims 1 to 5, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, 20. The αvβ6 integrin ligand according to any one of claims 18 to 19, wherein The targeting portion comprises a group formed by removing a hydrogen atom from the compound represented by formula I-1 or its stereoisomer or a pharmaceutically acceptable salt thereof, and the linker portion is The linker moiety is covalently linked to the targeting moiety, X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b 、R A1 、R A2 、R A3 , Q are each independently as defined in claim 6, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10; Wherein, the targeting portion comprises a compound represented by formula I-1 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in R 3 or Above (preferably A group formed by removing a hydrogen atom; Preferably, the αvβ6 integrin ligand comprises a group represented by formula I-1b, wherein X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b 、R A1 、R A2 、R A3 , Q are each independently as defined in claim 6, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, 21. The αvβ6 integrin ligand according to any one of claims 18 to 19, wherein The targeting portion comprises a group formed by removing a hydrogen atom from the compound of formula I-2 or its stereoisomer or a pharmaceutically acceptable salt thereof, and the linker portion is The linker moiety is covalently linked to the targeting moiety, n、n2、R 1 、R 2 、R 3 , Ring A are each independently as defined in claim 7, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10; Wherein, the targeting portion comprises a compound represented by formula I-2 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in R 3 or Above (preferably A group formed by removing a hydrogen atom; Preferably, the αvβ6 integrin ligand comprises a group shown in formula I-2b, wherein n, n2, R 1 、R 2 、R 3 , Ring A are each independently as defined in claim 7, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, 22. The αvβ6 integrin ligand according to any one of claims 18 to 19, wherein The targeting portion comprises a group formed by removing a hydrogen atom from the compound of formula I-3 or its stereoisomer or a pharmaceutically acceptable salt thereof, and the linker portion is The linker moiety is covalently linked to the targeting moiety, n、n2、R 1 、R 2 、R 3 、R A1 、R A2 、R A3 , L are each independently as defined in claim 8, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10; Wherein, the targeting portion comprises a compound represented by formula I-3 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in R 3 or Above (preferably A group formed by removing a hydrogen atom; Preferably, the αvβ6 integrin ligand comprises a group shown in formula I-3b, wherein n, n2, R 1 、R 2 、R 3 、R A1 、R A2 、R A3 , L are each independently as defined in claim 8, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, 23. The αvβ6 integrin ligand according to any one of claims 18 to 19, wherein The targeting portion comprises a group formed by removing a hydrogen atom from the compound of formula I-4 or its stereoisomer or a pharmaceutically acceptable salt thereof, and the linker portion is The linker moiety is covalently linked to the targeting moiety, R 1 、R 2 、R 3 、R 5a 、R 5b , L0, and A ring are each independently as defined in claim 9, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10; Wherein, the targeting portion comprises a compound represented by formula I-4 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in R 3 or Above (preferably A group formed by removing a hydrogen atom; Preferably, the αvβ6 integrin ligand comprises a group represented by formula I-4b, R 1 、R 2 、R 3 、R 5a 、R 5b , L0, and A ring are each independently as defined in claim 9, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, 24. The αvβ6 integrin ligand according to any one of claims 18 to 23, wherein The αvβ6 integrin ligand comprises a group selected from the group consisting of:

25. A delivery conjugate targeting αvβ6 integrin, comprising: an αvβ6 integrin ligand, and a group comprising a molecule to be delivered, wherein the αvβ6 integrin ligand and the group comprising a molecule to be delivered are covalently linked, wherein: The αvβ6 integrin ligand is as defined in any one of claims 11-24.

26. The delivery conjugate targeting αvβ6 integrin according to claim 25, wherein The αvβ6 integrin ligand comprises a targeting portion and a linker portion, wherein the targeting portion comprises a group formed by removing a hydrogen atom from a compound represented by Formula I or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and the linker portion is The linker moiety is covalently linked to the targeting moiety and the group comprising the molecule to be delivered, X, Y, n, m, R 1 、R 2 、R 3 、R 4 、R 5a 、R 5b , Ring A, Ring B, Q, and L1 are each independently as defined in any one of claims 1 to 5, and n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10; Wherein, the targeting portion comprises a compound of formula I or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in R 3 、R 4 or a group formed by removing a hydrogen atom from the B ring (preferably from the B ring); Preferably, Ring B is When the delivery conjugate targeting αvβ6 integrin comprises a compound represented by formula Ic, X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b , Ring A, Q, L1 are each independently as defined in any one of claims 1 to 5, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, Z is a group comprising a molecule to be delivered, 27. The delivery conjugate targeting αvβ6 integrin according to any one of claims 25-26, wherein The delivery conjugate targeting αvβ6 integrin comprises a compound shown in formula I-1c, X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b 、R A1 、R A2 、R A3 , Q are each independently as defined in claim 6, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, Z is a group comprising a molecule to be delivered, 28. The delivery conjugate targeting αvβ6 integrin according to any one of claims 25-26, wherein The delivery conjugate targeting αvβ6 integrin comprises a compound shown in formula I-2c, wherein n, n2, R 1 、R 2 、R 3 , Ring A are each independently as defined in claim 7, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, Z is a group comprising a molecule to be delivered, 29. The delivery conjugate targeting αvβ6 integrin according to any one of claims 25-26, wherein The delivery conjugate targeting αvβ6 integrin comprises a compound shown in formula I-3c, wherein n, n2, R 1 、R 2 、R 3 、R A1 、R A2 、R A3 , L are each independently as defined in claim 8, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, Z is a group comprising a molecule to be delivered, 30. The delivery conjugate targeting αvβ6 integrin according to any one of claims 25-26, wherein The delivery conjugate targeting αvβ6 integrin comprises a compound shown in formula I-4c, R 1 、R 2 、R 3 、R 5a 、R 5b , L0, and A ring are each independently as defined in claim 9, n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10, and Z is a group containing a molecule to be delivered, 31. The αvβ6 integrin targeted delivery conjugate of any one of claims 25-30, wherein The delivery conjugate targeting αvβ6 integrin is selected from: Each Z independently represents the group comprising the molecule to be delivered.

32. The αvβ6 integrin targeted delivery conjugate of any one of claims 25-31, wherein The group comprising the delivered molecule further comprises a linker group, wherein the linker group is covalently linked to the αvβ6 integrin ligand and the delivered molecule; Preferably, the linker group is in: L a1 Selected from p1 is selected from 1, 2, 3, 4, 5, preferably 3; preferably, L a1 One side of the carbonyl end is covalently linked to the molecule being delivered, and the other side of the carbonyl end is covalently linked to L a2 covalent attachment; L a2 Selected from p2 is selected from 1, 2, 3, 4, 5, preferably 2; preferably, L a2 The amino terminus of L a1 Covalently linked, carbonyl end and L a3 covalent attachment; L a3 Selected from Each p3 is independently selected from 1, 2, 3, 4, 5, preferably 2; preferably, L a3 The amino terminus of L a2 Covalently linked, the triazole N-atom end or the triazole C-atom end is covalently linked to the αvβ6 integrin ligand; More preferably, the linker group is selected from The carbonyl end is covalently linked to the delivered molecule, and the triazole N-atom end or the triazole C-atom end is covalently linked to the αvβ6 integrin ligand.

33. The αvβ6 integrin targeted delivery conjugate of any one of claims 25-32, wherein The delivered molecule includes, but is not limited to, an RNAi agent, a small molecule, an antibody, an antibody fragment, an immunoglobulin, a monoclonal antibody, a label or marker, a lipid, a natural or modified nucleic acid, a natural or modified nucleic acid oligonucleotide, a natural or modified nucleic acid polynucleotide, a peptide, an aptamer, a polymer, a polyamine, a protein, a toxin, a vitamin, polyethylene glycol, a hapten, digoxigenin, biotin, a radioactive atom or molecule, or a fluorophore; Preferably, the delivered molecule is an RNAi agent; Preferably, the delivered molecule is a small interfering RNA (siRNA), the siRNA comprises a sense strand and an antisense strand, and the sequences of the sense strand and antisense strand of the siRNA are selected from the sense strand and antisense strand sequences of any duplex 1 to duplex 3 described in Table 6.

34. The αvβ6 integrin targeted delivery conjugate of any one of claims 25-33, wherein The delivery conjugate targeting αvβ6 integrin is selected from: Each Z R independently represent the molecules to be delivered, preferably, each Z R RNAi agents are represented independently.

35. An αvβ6 integrin ligand precursor comprising a targeting moiety as defined in any one of claims 11 to 17, and a reactive group for conjugation to a group comprising a molecule to be delivered.

36. The αvβ6 integrin ligand precursor according to claim 35, wherein The reactive group is selected from an azide group and an alkyne group.

37. The αvβ6 integrin ligand precursor according to any one of claims 35 to 36, wherein The αvβ6 integrin ligand precursor further comprises a linker moiety covalently linked to the targeting moiety and the reactive group; Preferably, the linker moiety comprises a PEG linker; Preferably, the linker portion is n1 is selected from integers of 2 to 20, preferably from integers of 2 to 10.

38. An αvβ6 integrin ligand precursor, which is a compound represented by formula Id or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, in: X, Y, n, m, R 1 、R 2 、R 3 、R 4 、R 5a 、R 5b , Ring A, Ring B, Q, and L1 are each independently as defined in any one of claims 1 to 5; L2 is n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10; R 6 It is selected from an azido group and an alkynyl group, and is preferably an azido group (-N3).

39. The αvβ6 integrin ligand precursor according to claim 38, which is a compound represented by formula I-1d or a stereoisomer thereof or a pharmaceutically acceptable salt thereof. in: X, Y, n, m, R 1 、R 2 、R 3 、R 5a 、R 5b 、R A1 、R A2 、R A3 , Q are each independently as defined in claim 6; n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10; R 6 It is selected from an azido group and an alkynyl group, and is preferably an azido group (-N3).

40. The αvβ6 integrin ligand precursor of claim 38, which is a compound represented by formula I-2d or a stereoisomer thereof or a pharmaceutically acceptable salt thereof. in: n、n2、R 1 、R 2 、R 3 , Ring A are each independently as defined in claim 7; n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10; R 6 It is selected from an azido group and an alkynyl group, and is preferably an azido group (-N3).

41. The αvβ6 integrin ligand precursor of claim 38, which is a compound represented by formula I-3d or a stereoisomer thereof or a pharmaceutically acceptable salt thereof. in: n、n2、R 1 、R 2 、R 3 、R A1 、R A2 、R A3 , L are each independently as defined in claim 8; n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10; R 6 It is selected from an azido group and an alkynyl group, and is preferably an azido group (-N3).

42. The αvβ6 integrin ligand precursor of claim 38, which is a compound represented by formula I-4d or a stereoisomer thereof or a pharmaceutically acceptable salt thereof. in: R 1 、R 2 、R 3 、R 5a 、R 5b , L0, and A ring are each independently as defined in claim 9; n1 is selected from an integer of 2 to 20, preferably an integer of 2 to 10; R 6 It is selected from an azido group and an alkynyl group, and is preferably an azido group (-N3).

43. The αvβ6 integrin ligand precursor according to any one of claims 38 to 42, which is a compound selected from the group consisting of:

44. A linker compound, which is a compound represented by Formula IIp or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, in: L a1 Selected from p1 is selected from 1, 2, 3, 4, 5, preferably 3; preferably, L a1 The carbonyl end on one side is covalently linked to the hydroxyl group, and the carbonyl end on the other side is covalently linked to L a2 covalent attachment; L a2 Selected from p2 is selected from 1, 2, 3, 4, 5, preferably 2; preferably, L a2 The amino terminus of L a1 Covalently linked, carbonyl end and L a3p covalent attachment; L a3p Selected from p3 is selected from 1, 2, 3, 4, 5, preferably 2; preferably, L a3p The amino terminus of L a2 Covalently linked.

45. The linker compound of claim 44, wherein The linker compound is 46. ​​A composition or pharmaceutical composition comprising the compound of any one of claims 1-10, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the αvβ6 integrin ligand of any one of claims 11-24, or the αvβ6 integrin targeted delivery conjugate of any one of claims 25-34, or the αvβ6 integrin ligand precursor of any one of claims 35-43, or the linker compound of any one of claims 44-45; and optionally a pharmaceutically acceptable excipient.

47. Use of the compound of any one of claims 1-10, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the αvβ6 integrin ligand of any one of claims 11-24, or the αvβ6 integrin targeted delivery conjugate of any one of claims 25-34, or the αvβ6 integrin ligand precursor of any one of claims 35-43, or the linker compound of any one of claims 44-45, or the composition or pharmaceutical composition of claim 46 in the preparation of an agent or medicament for delivering a target molecule.

48. Use of the compound of any one of claims 1-10, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the αvβ6 integrin ligand of any one of claims 11-24, or the αvβ6 integrin targeted delivery conjugate of any one of claims 25-34, or the αvβ6 integrin ligand precursor of any one of claims 35-43, or the linker compound of any one of claims 44-45, or the composition or pharmaceutical composition of claim 46 in the preparation of a medicament for treating and / or preventing a disease.