Isoindoline compound containing phenylethoxyalkyleneamine structure, conjugate thereof, and use thereof

Abstract

The present invention relates to an isoindoline compound containing a phenylethoxyalkyleneamine structure, a conjugate thereof, and use thereof. The isoindoline compound containing a phenylethoxyalkyleneamine structure has a structure represented by formula (I). The conjugate thereof has a structure represented by formula (B). The compound of the present invention exhibits increased proliferation inhibition activity for cancer cells and a degradation effect on proteins such as GSPT1, demonstrating practical values.

Description

Isoindoline compounds containing phenylethoxyalkyleneamine structure, their couplings and uses

[0001] This application claims priority to the following Chinese patent applications filed on October 11, 2024, with application number 202411419844.X, entitled "Isoindoline compounds containing a phenylethoxyalkyleneamine structure, their couplings and uses thereof"; filed on March 24, 2025, with application number 202510353109.1, entitled "Isoindoline compounds containing a phenylethoxyalkyleneamine structure, their couplings and uses thereof"; and filed on August 29, 2025, with application number 202511232966.2, entitled "Isoindoline compounds containing a phenylethoxyalkyleneamine structure, their couplings and uses thereof", the entire contents of which are incorporated herein by reference. Technical Field

[0002] This invention relates to the field of pharmaceutical technology, and in particular to an isoindoline compound containing a phenylethoxyalkyleneamine structure, an antibody-drug conjugate containing an isoindoline compound containing a phenylethoxyalkyleneamine structure, a pharmaceutical composition thereof, a method of preparation thereof, and therapeutic uses thereof. Background Technology

[0003] Eukaryotic release factor 3a (eRF3a), encoded by the G1 to S phase transition 1 (GSPT1) gene, is upregulated in various human cancers and is a potential proto-oncogene. As a key member of the peptide release factor family, it participates in multiple biological processes, including terminating protein translation, regulating intracellular mRNA degradation, regulating cell cycle and apoptosis, and participating in cytoskeleton formation. Recent evidence suggests that GSPT1 is abnormally highly expressed in various malignant tumors and plays an important role in tumorigenesis and development. GSPT1 is a carcinogenic driver in several cancer types, including hematologic malignancies, breast cancer, liver cancer, gastric cancer, and prostate cancer. Furthermore, GSPT1 is significantly upregulated in cancer tissues and cell lines; its high expression is positively correlated with tumor size, and its overexpression can promote cancer cell proliferation and metastasis.

[0004] Several publications have reported GSPT1 degraders with different structures, including WO2023070120A1, WO2023201282 A1, WO2023274246A1, and WO2023069708 A1. The activity and selectivity of these existing compounds still need further improvement. Therefore, developing GSPT1 degraders with higher activity and selectivity to further improve the therapeutic effect of tumors and reduce the toxic side effects of drugs has significant research value and practical significance. Furthermore, developing antibody-drug conjugates (ADCs) based on GSPT1 small molecule degraders that can bind to tumor cells and cleave to release bioactive molecules in the tumor microenvironment to improve efficacy and eliminate or reduce the toxic side effects caused by small molecule degraders acting on non-disease tissues is of great importance. Summary of the Invention

[0005] One object of the present invention is to provide a class of novel isoindoline compounds containing phenylethoxyalkyleneamine structures and their compositions, which, as cereblon regulators, exhibit enhanced proliferative inhibitory activity against cancer cells and degradation of proteins such as GSPT1.

[0006] Another object of the present invention is to provide a class of antibody-drug conjugates (ADCs) using isoindoline compounds with a phenylethoxyalkyleneamine structure as drug molecules, as well as their compositions, preparation methods, and uses. These ADCs combine the tumor-targeting activity of antibodies with the high activity of bioactive compounds, exhibiting low aggregation and high inhibitory activity against tumor cells, demonstrating promising therapeutic and safety advantages.

[0007] The first aspect of the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0008] in,

[0009] R1 is hydrogen or halogen;

[0010] R2 and R3 are each independently hydrogen or C. 1-8 Alkyl (preferably C) 1-6 Alkyl, more preferably C 1-3 alkyl);

[0011] Furthermore, R1, R2, and R3 are not all hydrogen at the same time;

[0012] n is 1, 2, or 3;

[0013] R 41 R 42 R 43 R 44 Each can be either hydrogen or halogen;

[0014] R5, R6, R a R b R c R d R e R f m is selected from the following group of definitions:

[0015] (i) R5 and R6 are each independently hydrogen and C. 1-8 Alkyl, C 3-6 cycloalkyl, -C 1-4 Alkylene-C 3-6 cycloalkyl, halogenated C 1-8 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-4 Alkylene-R7, the C 1-8 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-8 alkyl;

[0016] R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-4 Alkyl, -S(O)-C 1-4 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens;

[0017] R a R b R c R d R e R f Each independently is hydrogen, C 1-8 Alkyl or halogen;

[0018] m is 0, 1, or 2;

[0019] (ii) R6 is hydrogen, C 1-8 Alkyl, C 3-6 cycloalkyl, -C 1-4 Alkylene-C 3-6 cycloalkyl, halogenated C 1-8 Alkyl, or 4- to 6-membered heterocyclic alkyl, wherein the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-8 alkyl;

[0020] R a R b R d Re R f Each independently is hydrogen, C 1-8 Alkyl or halogen;

[0021] R c The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens;

[0022] m is 0, 1, or 2;

[0023] (iii) R6 is hydrogen, C 1-8 Alkyl, C 3-6 cycloalkyl, -C 1-4 Alkylene-C 3-6 cycloalkyl, halogenated C 1-8 Alkyl, or 4- to 6-membered heterocyclic alkyl, wherein the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-8 alkyl;

[0024] R a R b R d R c R f Each independently is hydrogen, C 1-8 Alkyl or halogen;

[0025] R e The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens;

[0026] m can be 0, 1, or 2.

[0027] In some implementations, R1 is a halogen; R2 and R3 are each independently hydrogen.

[0028] In some implementations, R1 is fluorine; R2 and R3 are each independently hydrogen.

[0029] In some implementations, R1 is hydrogen; R2 is hydrogen; and R3 is C. 1-3 alkyl.

[0030] In some implementations, R5 is hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3Alkyl (preferably fluorinated C) 1-3 Alkyl), 4- to 6-membered heterocyclic alkyl or -C 1-2 Alkylene-R7, the C 1-3 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl;

[0031] R6 is hydrogen;

[0032] R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens;

[0033] R a R b R c R d R e R f Each independently is hydrogen, C 1-3 Alkyl or halogen;

[0034] m is 0.

[0035] In some implementations, R6 is hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl, or 4- to 6-membered heterocyclic alkyl, wherein the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl;

[0036] R a R b R d R e R f Each independently is hydrogen, C 1-3 Alkyl or halogen;

[0037] R c The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens;

[0038] m is 0 or 1.

[0039] In some implementations, R6 is hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl, or 4- to 6-membered heterocyclic alkyl, wherein the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl;

[0040] R a R b R d R c R f Each independently is hydrogen, C 1-3 Alkyl or halogen;

[0041] R e The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens;

[0042] m is 0.

[0043] In some embodiments, the 4- to 6-membered heterocyclic alkyl group is selected from: azahexacyclic butyl, oxacyclobutyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, oxazolidinyl, dioxolane, piperidinyl, piperazinyl, morpholinyl, dioxahexacyclic, thiomorpholinyl, thiomorpholin-1,1-dioxide, tetrahydropyranyl, pyrrolidine-2-one, dihydrofuran-2(3H)-one, morpholin-3-one, piperazin-2-one, and piperidin-2-one.

[0044] In some embodiments, the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is selected from: aziridine, tetrahydropyrrolyl, oxazolyl, piperidinyl, piperazine, morpholinyl, thiomorpholinyl, thiomorpholin-1,1-dioxide, pyrrolidine-2-one, morpholin-3-one, piperazine-2-one, and piperidin-2-one.

[0045] In some embodiments, the compound represented by formula (I) has the structure shown in formula (IA) or formula (IB):

[0046] Where R 51 R 62 Each independently is hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-2 Alkylene-R7, the C 1-3 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl;

[0047] R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens;

[0048] n is 1;

[0049] s is 1, 2, or 3;

[0050] t is 0, 1, or 2;

[0051] And when s is 1, t is not 0;

[0052] R0 is independently hydrogen and C. 1-8 Alkyl groups, halogens;

[0053] q can be 0, 1, 2, or 3;

[0054] p is 0, 1, or 2.

[0055] In some implementations, R1 is fluorine; R2 and R3 are each independently hydrogen.

[0056] In some implementations, R1 is hydrogen; R2 is hydrogen; and R3 is C. 1-3 alkyl.

[0057] In some implementations, R1 is hydrogen; R2 is hydrogen; and R3 is methyl.

[0058] In some implementation schemes, R 51 Ethyl, n-propyl, isopropyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 Cycloalkyl, 4- to 6-membered heterocycloalkyl or -C 1-2 Alkylene-R7; the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl;

[0059] R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens.

[0060] In some implementation schemes, R 51 Groups selected from the following group: methyl, -CH2-CF3.

[0061] In some implementation schemes, R 51 Groups selected from the following group: -CH2-CN, -(CH2)2-S(O)2-CH3, -(CH2)2-OH, methyl, ethyl, n-propyl, isopropyl, -CH2-CF3, -CH2-CHF2, -(CH2)2-S(O)2-NH2, -CH(CF3)-CH3, -CH2-C(O)NH2, -(CH2) 0-1 -Halocyclopropyl (preferably -(CH2)) 0-1 -Fluorocyclopropyl), -(CH2) 0-1 -Halogenated cyclobutyl (preferably -(CH2)) 0-1 -Fluorocyclobutyl), -(CH2) 0-1 -Halocyclopentyl (preferably -(CH2)) 0-1 -Fluorocyclopentyl), -(CH2) 0-1 -Halogenated cyclohexyl (preferably -(CH2)) 0-1 -Fluorocyclohexyl), -(CH2) 0-1 -Cyclopropyl, -(CH2) 0-1 -cyclobutyl, -(CH2) 0-1 -cyclopentyl, -(CH2) 0-1 - Cyclohexyl, aziridine, oxaziridine, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrroleyl, oxazolidinyl, dioxopentyl, piperidinyl, piperazine, morpholinyl, dioxohexyl, thiomorpholinyl, thiomorpholin-1,1-dioxide, tetrahydropyranyl, thiocyclobutane-1,1-dioxide, tetrahydrothiophene-1,1-dioxide.

[0062] In some implementation schemes, R 62 It is hydrogen.

[0063] In some implementations, in formula (IA), R 51 C 3-6 cycloalkyl or halogenated C 1-3 Alkyl, the C 3-6The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of: halogens; n is 1.

[0064] In some implementations, in formula (IA), R 51 The group selected from the following group: -CH2-CF3, -CH2-CHF2, cyclopropyl, fluorocyclobutyl, -CH(CF3)-CH3.

[0065] In some embodiments, the compound of formula (I) is selected from any of the following structures:

[0066] In some embodiments, the compound of formula (I) is selected from any of the following structures:

[0067] In some embodiments, the compound of formula (I) is selected from any of the following structures:

[0068] The compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, described in the first aspect of this invention has significant protein regulatory effects and enhanced cell proliferation inhibition effects, said proteins including GSPT1, N-MYC, and C-MYC, and is a protein regulator to restore protein homeostasis, including GSPT1 activity.

[0069] A second aspect of the present invention provides a compound of formula (II), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0070] Where R1' is hydrogen or halogen;

[0071] R2' and R3' are each independently hydrogen or C. 1-8 Alkyl (preferably C) 1-6 Alkyl, more preferably C 1-3 alkyl);

[0072] n is 1, 2, or 3;

[0073] R 41 R 42 R 43 R 44 Each can be either hydrogen or halogen;

[0074] R5', R6, R a R b R c R d R e R f m is selected from the following group of definitions:

[0075] (i)R5' is C 3-6 cycloalkyl, -C 1-4 Alkylene-C 3-6 cycloalkyl, halogenated C 1-8 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-4 Alkylene-R7, the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-8 alkyl;

[0076] R6 is hydrogen, C 1-8 Alkyl, C 3-6 cycloalkyl, -C 1-4 Alkylene-C 3-6 cycloalkyl, halogenated C 1-8 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-4 Alkylene-R7, the C 1-8 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-8 alkyl;

[0077] R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-4 Alkyl, -S(O)-C 1-4 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens;

[0078] R a R b R c R d R e R f Each independently is hydrogen, C 1-8 Alkyl or halogen;

[0079] m is 0, 1, or 2;

[0080] (ii) R6 is hydrogen, C 1-8 Alkyl, C 3-6 cycloalkyl, -C 1-4 Alkylene-C 3-6 cycloalkyl, halogenated C 1-8 Alkyl, or 4- to 6-membered heterocyclic alkyl, wherein the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-8 alkyl;

[0081] R a R b R d R e R f Each independently is hydrogen, C 1-8 Alkyl or halogen;

[0082] R c The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5'; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens;

[0083] m is 0, 1, or 2;

[0084] (iii) R6 is hydrogen, C 1-8 Alkyl, C 3-6 cycloalkyl, -C 1-4 Alkylene-C 3-6 cycloalkyl, halogenated C 1-8 Alkyl, or 4- to 6-membered heterocyclic alkyl, wherein the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-8 alkyl;

[0085] R a R b R d R c R f Each independently is hydrogen, C 1-8 Alkyl or halogen;

[0086] R e The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5'; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens;

[0087] m can be 0, 1, or 2.

[0088] In some implementations, R1', R2', and R3' are hydrogen.

[0089] In some implementations, R5' is C 3-6 cycloalkyl, -C 1-4 Alkylene-C 3-6 cycloalkyl, halogenated C 1-8 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-4 Alkylene-R7, the C 1-8 Alkyl, C 3-6Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-8 alkyl;

[0090] R6 is hydrogen;

[0091] R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-4 Alkyl, -S(O)-C 1-4 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens;

[0092] R a R b R c R d R e R f Each independently is hydrogen, C 1-8 Alkyl or halogen;

[0093] m can be 0, 1, or 2.

[0094] In some implementations, R6 is hydrogen.

[0095] In some embodiments, the compound represented by formula (II) has the structure shown in formula (II-A) or formula (II-B):

[0096] Where R 51 'For C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-2 Alkylene-R7, the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl;

[0097] R 62 Each independently is hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-2 Alkylene-R7, the C 1-3 Alkyl, C 3-6Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl;

[0098] R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens;

[0099] n is 1;

[0100] s is 1, 2, or 3;

[0101] t is 0, 1, or 2;

[0102] And when s is 1, t is not 0;

[0103] R0 is independently hydrogen and C. 1-8 Alkyl groups, halogens;

[0104] q can be 0, 1, 2, or 3;

[0105] p is 0, 1, or 2.

[0106] In some implementation schemes, R 51 The following groups are selected: -CH2-CN, -(CH2)2-S(O)2-CH3, methyl, -CH2-CF3, -CH2-CHF2, -(CH2)2-S(O)2-NH2, -CH(CF3)-CH3, -CH2-C(O)NH2, -(CH2) 0-1 -Halocyclopropyl (preferably -(CH2)) 0-1 -Fluorocyclopropyl), -(CH2) 0-1 -Halogenated cyclobutyl (preferably -(CH2)) 0-1 -Fluorocyclobutyl), -(CH2) 0-1 -Halocyclopentyl (preferably -(CH2)) 0-1 -Fluorocyclopentyl), -(CH2) 0-1 -Halogenated cyclohexyl (preferably -(CH2)) 0-1 -Fluorocyclohexyl), -(CH2) 0-1 -Cyclopropyl, -(CH2) 0-1 -cyclobutyl, -(CH2) 0-1 -cyclopentyl, -(CH2) 0-1- Cyclohexyl, azirhexacyclobutyl, oxacyclobutyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, oxazolidinyl, dioxopranyl, piperidinyl, piperazine, morpholinyl, dioxohexacycloyl, thiomorpholinyl, thiomorpholin-1,1-dioxide, tetrahydropyranyl.

[0107] In some implementation schemes, R 62 It is hydrogen.

[0108] In some implementations, in formula (II-A), R 51 'For C 3-6 cycloalkyl or halogenated C 1-3 Alkyl, the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of: halogens; n is 1.

[0109] In some implementations, in formula (II-A), R 51 The group selected from the following group: -CH2-CF3, -CH2-CHF2, cyclopropyl, fluorocyclobutyl, -CH(CF3)-CH3.

[0110] A third aspect of the present invention provides a compound of formula (A), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0111] in,

[0112] T is a linker group containing a functional group that can react with a thiol group;

[0113] L is a linking group containing either non-cleavable or cleavable units;

[0114] R1 is hydrogen or halogen;

[0115] R2 and R3 are each independently hydrogen or C. 1-8 Alkyl (preferably C) 1-6 Alkyl, more preferably C 1-3 alkyl);

[0116] Furthermore, R1, R2, and R3 are not all hydrogen at the same time;

[0117] n is 1, 2, or 3;

[0118] R 41 R 42 R 43 R 44 Each can be either hydrogen or halogen;

[0119] R5, R a R b R c R d Re R f m is selected from the following group of definitions:

[0120] (i) R5 is hydrogen, C 1-8 Alkyl, C 3-6 cycloalkyl, -C 1-4 Alkylene-C 3-6 cycloalkyl, halogenated C 1-8 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-4 Alkylene-R7, the C 1-8 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-8 alkyl;

[0121] R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-4 Alkyl, -S(O)-C 1-4 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens;

[0122] R a R b R c R d R e R f Each independently is hydrogen, C 1-8 Alkyl or halogen;

[0123] m is 0, 1, or 2;

[0124] (ii)R a R b R d R e R f Each independently is hydrogen, C 1-8 Alkyl or halogen;

[0125] R c The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens;

[0126] m is 0, 1, or 2;

[0127] (iii)R a R b R d R c Rf Each independently is hydrogen, C 1-8 Alkyl or halogen;

[0128] R e The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens;

[0129] m can be 0, 1, or 2.

[0130] In some embodiments, L is a linker group containing a non-cleavable unit. In some embodiments, L is a linker group containing a cleavable unit.

[0131] In some embodiments, the compound represented by formula (A) has the structure shown in formula (AA) or formula (AB):

[0132] Where R 51 For hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-2 Alkylene-R7, the C 1-3 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl;

[0133] R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens;

[0134] n is 1;

[0135] s is 1, 2, or 3;

[0136] t is 0, 1, or 2;

[0137] And when s is 1, t is not 0;

[0138] R0 is independently hydrogen and C. 1-8 Alkyl groups, halogens;

[0139] q can be 0, 1, 2, or 3;

[0140] p is 0, 1, or 2.

[0141] In some implementations, R1 is fluorine; R2 and R3 are each independently hydrogen.

[0142] In some implementations, R1 is hydrogen; R2 is hydrogen; and R3 is C. 1-3 alkyl.

[0143] In some implementations, R1 is hydrogen; R2 is hydrogen; and R3 is methyl.

[0144] In some implementation schemes, R 51 Ethyl, n-propyl, isopropyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 Cycloalkyl, 4- to 6-membered heterocycloalkyl or -C 1-2 Alkylene-R7; the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl;

[0145] R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens.

[0146] In some implementation schemes, R 51 Groups selected from the following group: methyl, -CH2-CF3.

[0147] In some implementation schemes, R 51 Groups selected from the following group: -CH2-CN, -(CH2)2-S(O)2-CH3, -(CH2)2-OH, methyl, ethyl, n-propyl, isopropyl, -CH2-CF3, -CH2-CHF2, -(CH2)2-S(O)2-NH2, -CH(CF3)-CH3, -CH2-C(O)NH2, -(CH2) 0-1 -Halocyclopropyl (preferably -(CH2)) 0-1 -Fluorocyclopropyl), -(CH2) 0-1 -Halogenated cyclobutyl (preferably -(CH2)) 0-1 -Fluorocyclobutyl), -(CH2) 0-1 -Halocyclopentyl (preferably -(CH2)) 0-1-Fluorocyclopentyl), -(CH2) 0-1 -Halogenated cyclohexyl (preferably -(CH2)) 0-1 -Fluorocyclohexyl), -(CH2) 0-1 -Cyclopropyl, -(CH2) 0-1 -cyclobutyl, -(CH2) 0-1 -cyclopentyl, -(CH2) 0-1 - Cyclohexyl, aziridine, oxaziridine, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrroleyl, oxazolidinyl, dioxopentyl, piperidinyl, piperazine, morpholinyl, dioxohexyl, thiomorpholinyl, thiomorpholin-1,1-dioxide, tetrahydropyranyl, thiocyclobutane-1,1-dioxide, tetrahydrothiophene-1,1-dioxide.

[0148] In some implementations, T stands for T1-T2-; where T1 is a functional group selected from the following group:

[0149] Where X is a halogen;

[0150] T2 is absent or is a linking group selected from the following group:

[0151] The waveform line represents the connection point with T1, the asterisk represents the connection point with L, and t1 is an integer from 1 to 10; t2 and t3 are each an independent integer from 0 to 10.

[0152] In some implementations, T is selected from

[0153] A fourth aspect of the present invention provides a conjugate of formula (B), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0154] Where Ab represents an antibody or antigen-binding fragment;

[0155] y can be any value between 1 and 10;

[0156] S represents the sulfur atom of the thiol group in the Ab portion;

[0157] T1' is selected from:

[0158] The wavy line indicates the connection point with T2, and the asterisk indicates the connection point with S; T2 is absent or is a linking group selected from the following groups:

[0159] The wavy line represents the connection point with T1', the asterisk represents the connection point with L, and t1 is an integer from 1 to 10; t2 and t3 are each an independent integer from 0 to 10.

[0160] L is a linking group containing either non-cleavable or cleavable units;

[0161] Other substituents are defined in compounds of formula (A).

[0162] In some implementations, Ab is an antibody against Her 2 or its antigen-binding fragment.

[0163] In some implementations, Ab is pertuzumab or trastuzumab.

[0164] In some implementations, y is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0165] In some embodiments, the drug-to-antibody ratio (DAR value) of the compound represented by formula (B) is any value between 1 and 10. In some embodiments, the DAR value is any value between 2 and 8. In some embodiments, the DAR value is any value between 2 and 6.

[0166] In some implementations, T1' is selected from The wavy line represents the connection point with T2, and the asterisk represents the connection point with S.

[0167] In some embodiments, the compound represented by formula (B) has the structure shown in formula (BA) or formula (BB):

[0168] Where R 51 For hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-2 Alkylene-R7, the C 1-3 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl;

[0169] R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens;

[0170] n is 1;

[0171] s is 1, 2, or 3;

[0172] t is 0, 1, or 2;

[0173] And when s is 1, t is not 0;

[0174] R0 is independently hydrogen and C. 1-8 Alkyl groups, halogens;

[0175] q can be 0, 1, 2, or 3;

[0176] p is 0, 1, or 2.

[0177] In some implementations, R1 is fluorine; R2 and R3 are each independently hydrogen.

[0178] In some implementations, R1 is hydrogen; R2 is hydrogen; and R3 is C. 1-3 alkyl.

[0179] In some implementations, R1 is hydrogen; R2 is hydrogen; and R3 is methyl.

[0180] In some implementation schemes, R 51 Ethyl, n-propyl, isopropyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 Cycloalkyl, 4- to 6-membered heterocycloalkyl or -C 1-2 Alkylene-R7; the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl;

[0181] R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens.

[0182] In some implementation schemes, R 51 Groups selected from the following group: methyl, -CH2-CF3.

[0183] In some implementation schemes, R 51Groups selected from the following group: -CH2-CN, -(CH2)2-S(O)2-CH3, -(CH2)2-OH, methyl, ethyl, n-propyl, isopropyl, -CH2-CF3, -CH2-CHF2, -(CH2)2-S(O)2-NH2, -CH(CF3)-CH3, -CH2-C(O)NH2, -(CH2) 0-1 -Halocyclopropyl (preferably -(CH2)) 0-1 -Fluorocyclopropyl), -(CH2) 0-1 -Halogenated cyclobutyl (preferably -(CH2)) 0-1 -Fluorocyclobutyl), -(CH2) 0-1 -Halocyclopentyl (preferably -(CH2)) 0-1 -Fluorocyclopentyl), -(CH2) 0-1 -Halogenated cyclohexyl (preferably -(CH2)) 0-1 -Fluorocyclohexyl), -(CH2) 0-1 -Cyclopropyl, -(CH2) 0-1 -cyclobutyl, -(CH2) 0-1 -cyclopentyl, -(CH2) 0-1 - Cyclohexyl, aziridine, oxaziridine, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrroleyl, oxazolidinyl, dioxopentyl, piperidinyl, piperazine, morpholinyl, dioxohexyl, thiomorpholinyl, thiomorpholin-1,1-dioxide, tetrahydropyranyl, thiocyclobutane-1,1-dioxide, tetrahydrothiophene-1,1-dioxide.

[0184] In some implementations, T1'-T2 are selected from...

[0185] The wavy line represents the connection point with L, and the asterisk represents the connection point with S.

[0186] In some implementations, L stands for L1-L2-L3; where

[0187] L1 either does not exist or is a structure selected from the following group:

[0188] The wavy line represents the connection point with L2, the asterisk represents the connection point with T or T2, and q1 is an integer from 1 to 8;

[0189] R L1 R L2 Each of the following is an independent structure selected from the group below:

[0190] Where p1 is an integer from 1 to 12; u is an integer from 1 to 15; s1, s2, s3, and s4 are each an integer from 1 to 15 independently; and v is an integer from 1 to 20.

[0191] R 1 R 2 Each independently is C 1-6 alkyl;

[0192] W does not exist or is -NH-C 1-3 alkylene-;

[0193] Ring A is a benzene ring or a 4- to 6-membered heterocyclic alkyl ring;

[0194] L2 is a short peptide composed of 2 to 4 amino acid residues; the amino acid residues are selected from natural amino acid residues and non-natural amino acid residues;

[0195] L3 either does not exist or has the following structure:

[0196] Where R L3 For hydrogen or

[0197] The wavy line represents the connection point with -N-, and the asterisk represents the connection point with L2.

[0198] In some embodiments, ring A is selected from: benzene ring, azacyclic butyl ring, tetrahydropyrrole ring, piperidine ring, piperazine ring.

[0199] In some implementation schemes, R 1 R 2 It is a methyl group.

[0200] In some embodiments, L2 is a dipeptide or tripeptide. In some embodiments, L2 is a dipeptide or tripeptide selected from the group consisting of: phenylalanine-lysine (-Phe-Lys-), valine-alanine (-Val-Ala-), valine-lysine (-Val-Lys-), alanine-lysine (-Ala-Lys-), valine-citrulline (-Val-Cit-), phenylalanine-citrulline (-Phe-Cit-), leucine-citrulline (-Leu-Cit-), and isoleucine-citrulline (-Ile-Cit-). The following are listed as potential adjuvants: phenylalanine-arginine (-Phe-Arg-), tryptophan-citrulline (-Trp-Cit-), glutamic acid-valine-alanine (-Glu-Val-Ala-), glutamic acid-valine-citrulline (-Glu-Val-Cit-), glycine-valine-citrulline (-Gly-Val-Cit-), alanine-alanine-alanine (-Ala-Ala-Ala-), and alanine-alanine-asparagine (-Ala-Ala-Asn-). In some embodiments, L2 is a dipeptide selected from the group consisting of: phenylalanine-lysine, valine-alanine, valine-lysine, alanine-lysine, and valine-citrulline. In some embodiments, L2 is a dipeptide selected from the group consisting of: phenylalanine-lysine, valine-citrulline, and valine-alanine.

[0201] In some implementations, L3 is selected from The wavy line represents the connection point with -N-, and the asterisk represents the connection point with L2.

[0202] In some implementations, TL is a structure selected from the following:

[0203] Where R L1 R L2 L2 and t1 are as defined in the instruction manual.

[0204] In some implementations, T1'-T2-L is a structure selected from the following:

[0205] Where R L1 R L2 L2 and t1 are as defined in the instruction manual; where the waveform line indicates the connection point with -N- and the asterisk indicates the connection point with S.

[0206] In some embodiments, L2 is a dipeptide selected from the group consisting of phenylalanine-lysine, valine-alanine, valine-lysine, alanine-lysine, and valine-citrulline.

[0207] In some implementations, L2 is The wavy line represents the connection point with L3, and the asterisk represents the connection point with L1.

[0208] In some implementation schemes, R L1 R L2 Each of the following is an independent structure selected from the group below:

[0209] Where p1 is an integer from 1 to 10; u is an integer from 1 to 15; s1, s2, s3, and s4 are each an integer from 1 to 15 independently; and v is an integer from 1 to 20.

[0210] R 1 R 2 Each independently is C 1-6 alkyl.

[0211] In some implementations, p1 is an integer from 5 to 10. In some implementations, p1 is 5, 6, 7, 8, 9, or 10.

[0212] In some implementations, s1 is an integer from 1 to 12. In some implementations, s1 is 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.

[0213] In some implementations, v is an integer from 1 to 10. In some implementations, v is 2, 3, 4, 5, 6, 7, or 8.

[0214] In some implementations, u is an integer from 1 to 12. In some implementations, u is 5, 6, 7, 8, 9, 10, 11, or 12.

[0215] In some implementations, TL is selected from the structure of Table A:

[0216] Table A

[0217] In some implementations, T1'-T2-L is selected from the structure in Table B:

[0218] Table B

[0219] The wavy line represents the connection point with -N-, and the asterisk represents the connection point with S. In some implementations, TL is selected from the following group of structures:

[0220] In some implementations, TL is In some implementations, T1'-T2-L is selected from the following group of structures: The wavy line represents the connection point with -N-, and the asterisk represents the connection point with S.

[0221] In some implementations, T1'-T2-L is The wavy line represents the connection point with -N-, and the asterisk represents the connection point with S.

[0222] In some implementations, t1 is an integer from 2 to 8. In some implementations, t1 is 3, 4, 5, or 6.

[0223] In some implementations, p1 is an integer from 5 to 10. In some implementations, p1 is 5, 6, 7, 8, 9, or 10.

[0224] In some implementations, s1 is an integer from 1 to 12. In some implementations, s1 is 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.

[0225] In some implementations, v is an integer from 1 to 10. In some implementations, v is 2, 3, 4, 5, 6, 7, or 8.

[0226] In some implementations, u is an integer from 1 to 12. In some implementations, u is 5, 6, 7, 8, 9, 10, 11, or 12.

[0227] In some implementations, TL is selected from the structure of Table C:

[0228] Table C

[0229] In some implementations, T1'-T2-L is selected from the structure of Table D:

[0230] Table D

[0231] The wavy lines represent the junctions with -N-, and the asterisks represent the junctions with S. In some embodiments, the compound represented by formula (A) is selected from the structures in Table E:

[0232] Table E

[0233] In some embodiments, the compound represented by formula (B) is selected from the structures in Table F:

[0234] Table F

[0235] Where Ab is the antibody against Her 2 or its antigen-binding fragment; y is any value between 1 and 10.

[0236] In some implementations, Ab is pertuzumab or trastuzumab.

[0237] In some embodiments, the compound represented by formula (B) is selected from any of the conjugates ADC01 to ADC19.

[0238] In some embodiments, the compound represented by formula (B) is selected from any of the conjugates ADC21 to ADC27.

[0239] In some embodiments, the compound represented by formula (B) is selected from any of the conjugates ADC28 to ADC36.

[0240] In some embodiments, the compound represented by formula (B) is selected from any of the conjugates ADC38 to ADC39.

[0241] In some embodiments, R1 is a halogen; R2 and R3 are each independently hydrogen. In some embodiments, R1 is fluorine; R2 and R3 are each independently hydrogen. In some embodiments, R1 is hydrogen; R2 is hydrogen; R3 is carbon. 1-3 alkyl.

[0242] In some implementations, R5 is hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl (preferably fluorinated C) 1-3 Alkyl), 4- to 6-membered heterocyclic alkyl or -C 1-2 Alkylene-R7, the C 1-3 Alkyl, C 3-6Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl;

[0243] R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens;

[0244] R a R b R c R d R e R f Each independently is hydrogen, C 1-3 Alkyl or halogen;

[0245] m is 0.

[0246] In some implementations, R5 is C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl (preferably fluorinated C) 1-3 alkyl) or -C 1-2 Alkylene-R7, the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens;

[0247] R7 is -S(O)2-C 1-3 Alkyl groups or -S(O)2-NH2;

[0248] R c R d R e R f Each is independently hydrogen;

[0249] m is 0.

[0250] In some implementation schemes, R a R b R d R e R f Each independently is hydrogen, C 1-3 Alkyl or halogen (preferably R) a R b R d R eR f (for hydrogen);

[0251] R c The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens;

[0252] m is 0 or 1.

[0253] In some embodiments, the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is azahexacyclic butyl, tetrahydropyrrole, or piperidinyl.

[0254] In some embodiments, in compounds of formula (AA) or formula (BA), the structure

[0255] Selected from the following structure:

[0256] In some embodiments, in compounds of formula (AB) or formula (BB), the structure

[0257] Selected from the following structure:

[0258] In some embodiments, the compounds represented by formula (A) or formula (B) have the following structure:

[0259] Selected from any specific structure of the compound of formula (I) of this application (preferably any specific structure of the compound of formula (I) in the examples), wherein Corresponding to any specific structure in the compound of formula (I) Partially, the wavy line represents the connection point of NH and L in the corresponding part of any specific structure in the compound of formula (I), and the asterisk indicates the connection with C(R). e R f ( ) connection points.

[0260] In some embodiments, the compounds represented by formula (A) or formula (B) have the following structure:

[0261] Selected from the following structure:

[0262] Another aspect of the present invention provides a method for preparing the compound of formula (B), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, the method comprising coupling Ab with the compound of formula (A);

[0263] Wherein, Ab and each group in formula (A) are defined as described in any of the schemes in this application.

[0264] In some embodiments, the method includes the step of coupling Ab with a compound of formula (A) in a suitable solvent and under suitable conditions to form a CS bond.

[0265] In some embodiments, the coupling reaction is carried out in water and / or an organic solvent. In some embodiments, the organic solvent is selected from N,N-dimethylformamide, dimethyl sulfoxide, N-methylpyrrolidone, nitriles (e.g., acetonitrile), alcohols (e.g., methanol, ethanol), or any combination thereof. In some embodiments, the method further includes a step of purifying the coupling product. In some embodiments, the coupling product is purified by chromatography.

[0266] A fifth aspect of the present invention provides a pharmaceutical composition comprising: the compound described in the first and second aspects of the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or

[0267] The compound described in the fourth aspect of this invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof;

[0268] And pharmaceutically acceptable carriers.

[0269] The sixth aspect of the present invention provides the use of the compounds of the first and second aspects of the present invention, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, the compounds of the fourth aspect of the present invention, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, and the pharmaceutical compositions of the fifth aspect of the present invention in the preparation of GSPT1 degrading agents.

[0270] The seventh aspect of the present invention provides the use of the compounds of the first and second aspects of the present invention, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, the compounds of the fourth aspect of the present invention, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, and the pharmaceutical compositions of the fifth aspect of the present invention in the preparation of medicaments for the treatment or prevention of diseases associated with mutations, expression imbalances, allosteric changes, and functional abnormalities of GSPT1, N-MYC, or C-MYC proteins.

[0271] In some implementations, the diseases include, but are not limited to, esophageal cancer, brain tumor, lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, urothelial carcinoma, non-Hodgkin's lymphoma, central nervous system tumors, prostate cancer, thyroid cancer, acute myeloid leukemia, and myelodysplastic syndrome.

[0272] In some embodiments, the compounds of the first and second aspects of the present invention, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, the compounds of the fourth aspect of the present invention, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, and the pharmaceutical compositions of the fifth aspect of the present invention are used to prepare medicaments for treating diseases related to or caused by GSPT1, particularly for treating cancers related to GSPT1, such as glioma, thyroid cancer, lung cancer, colorectal cancer, head and neck cancer, gastric cancer, liver cancer, pancreatic cancer, kidney cancer, urothelial carcinoma, prostate cancer, testicular cancer, breast cancer, cervical cancer, endometrial cancer, ovarian cancer, melanoma, multiple myeloma, acute myeloid leukemia, and myelodysplastic syndrome.

[0273] In some embodiments, the compounds of the first and second aspects of the present invention, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, the compounds of the fourth aspect of the present invention, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, and the pharmaceutical compositions of the fifth aspect of the present invention are used to treat solid tumors, particularly for the treatment of breast cancer.

[0274] The eighth aspect of the present invention provides a method for degrading GSPT1 protein in a patient in need, comprising administering to the patient a compound of the first or second aspect of the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, a compound of the fourth aspect of the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition of the fifth aspect of the present invention.

[0275] The present invention also provides compounds, pharmaceutically acceptable salts thereof, or stereoisomers thereof, as described in the first or second aspect of the present invention, for use as medicaments or for treatment; compounds, pharmaceutically acceptable salts thereof, or stereoisomers thereof, as described in the fourth aspect of the present invention; or pharmaceutical compositions as described in the fifth aspect of the present invention.

[0276] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Detailed Implementation

[0277] Through in-depth research, the inventors unexpectedly discovered a class of isoindoline compounds containing a phenylethoxyalkyleneamine structure, which can regulate the levels of proteins such as GSPT1 and exhibit significant selective inhibitory activity against IKZF1 and IKZF3 proteins. They show a significantly enhanced inhibitory effect on tumor cells, such as human breast ductal carcinoma cells, and can effectively alleviate or treat cancer and related diseases. Furthermore, based on this isoindoline compound containing a phenylethoxyalkyleneamine structure, antibody-drug conjugates with novel linker structures were further developed. Compared with existing conjugates with known linker groups, the conjugates of this invention have lower aggregation, exhibit higher stability, and show a significantly enhanced inhibitory effect on tumor cells, such as human breast ductal carcinoma cells. Therefore, the ADC and GSPT1 protein degrader of this invention have high drug development potential, and based on this, the inventors completed this invention.

[0278] Terminology Definition

[0279] To better understand the technical content of this invention, the terminology used in this invention will be further explained below. "alkyl" refers to a straight-chain or branched saturated aliphatic hydrocarbon group. "C 1-8 "Alkyl" refers to an alkyl group having 1 to 8 carbon atoms, preferably C14. 1-6 Alkyl, more preferably C 1-3 Alkyl; non-limiting examples of alkyl groups include: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and their various branched isomers.

[0280] "Cycloalkyl" and "cycloalkyl ring" are used interchangeably, both referring to a saturated monocyclic, bicyclic, or polycyclic cyclic hydrocarbon group that may be fused with an aryl or heteroaryl group. The cycloalkyl ring may optionally be substituted. In some embodiments, the cycloalkyl ring contains one or more carbonyl groups, such as oxo groups. "C" 3-8 "Cycloalkyl" refers to a monocyclic cycloalkyl group having 3 to 8 carbon atoms. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclobutanone, cyclopentanone, cyclopentane-1,3-dione, etc. Preferably C 3-6 Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0281] The terms "heterocyclic alkyl" and "heterocyclic alkyl ring" are used interchangeably and both refer to a cycloalkyl group containing at least one heteroatom selected from nitrogen, oxygen, and sulfur, which may be fused with an aryl or heteroaryl group. The heterocyclic alkyl ring may optionally be substituted. In some embodiments, the heterocyclic alkyl ring contains one or more carbonyl or thiocarbonyl groups, such as groups containing oxo and thio groups. "3- to 8-membered heterocyclic alkyl" refers to a monocyclic cyclic hydrocarbon group having 3 to 8 ring atoms, wherein 1, 2, or 3 ring atoms are heteroatoms selected from nitrogen, oxygen, and sulfur, preferably a 4- to 8-membered heterocyclic alkyl. More preferably, it is a 3- to 6-membered heterocyclic alkyl having 3 to 6 ring atoms, wherein 1 or 2 ring atoms are heteroatoms selected from nitrogen, oxygen, and sulfur. Even more preferably, it is a 4- to 6-membered heterocyclic alkyl having 4 to 6 ring atoms, wherein 1 or 2 ring atoms are heteroatoms selected from nitrogen, oxygen, and sulfur. Non-limiting embodiments include azircycloyl, ethylene oxide, azircyclobutyl, oxacyclobutyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, oxazolidinyl, dioxopentyl, piperidinyl, piperazinyl, morpholinyl, dioxohexyl, thiomorpholinyl, thiomorpholin-1,1-dioxide, tetrahydropyranyl, thiobutane-1,1-dioxide, tetrahydrothiophene-1,1-dioxide, azircyclobutane-2-keto, oxacyclobutane-2-keto, dihydrofuran-2(3H)-keto, pyrrolidine-2-keto, pyrrolidine-2,5-diketo, dihydrofuran-2,5-diketo, piperidin-2-keto, tetrahydro-2H-pyran-2-keto, piperazin-2-keto, morpholin-3-keto, etc.

[0282] "Heteroaryl" and "heteroaryl ring" are used interchangeably, both referring to a 4n+2 aromatic ring system (e.g., having 6 or 10 shared π electrons arranged in a ring) of monocyclic, bicyclic, or polycyclic aromatic rings having a ring carbon atom and a ring heteroatom, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur. The heteroaryl ring may optionally be substituted. "5 to 10-membered heteroaryl" refers to a monocyclic or bicyclic heteroaryl having 5 to 10 ring atoms, of which 1, 2, 3, or 4 ring atoms are heteroatoms. "5- to 6-membered heteroaryl" refers to a monocyclic heteroaryl group having 5 to 6 ring atoms, of which 1, 2, 3, or 4 ring atoms are heteroatoms. Non-limiting examples include thiophene, furanyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, pyrrole, pyrazolyl, triazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, tetrazolyl, isoxazolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, thiadiazolyl, pyridinyl, pyridinyl, triazinyl, and tetraazinyl. "9 or 10-membered heteroaryl" refers to a bicyclic heteroaryl group having 9 or 10 ring atoms, of which 1, 2, 3, or 4 ring atoms are heteroatoms. Non-limiting examples include indole, isoyindole, indazole, benzotriazolyl, benzothiophene, isobenzothiophene, benzofuranyl, benzoisofuranyl, benzimidazolyl, benzooxazolyl, benzoisooxazolyl, benzooxadiazolyl, benzothiazolyl, and benzoisothiazolyl. Azolyl, benzothiadiazolyl, indazinyl, purine, pyrido[3,2-d]pyrimidinyl, pyrido[2,3-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrido[4,3-d]pyrimidinyl, 1,8-naphthidyl, 1,7-naphthidyl, 1,6-naphthidyl, 1,5-naphthidyl, pteridyl, quinolinyl, isoquinolinyl, zolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. "Heteroatom" refers to nitrogen, oxygen, or sulfur. In heteroaryl groups containing one or more nitrogen atoms, the bonding point can be a carbon or nitrogen atom, provided the valence allows. Heteroaryl bicyclic systems can include one or more heteroatoms in one or both rings.

[0283] "Alkoxy" refers to -O-alkyl, where alkyl is defined as described above. Preferably C 1-8 Alkoxy, more preferably C 1-6 Alkoxy group, C is the most preferred. 1-3 Alkoxy groups. Non-limiting examples of alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, butoxy, tert-butoxy, isobutoxy, pentoxy, etc.

[0284] "A bond" refers to two groups connected by a covalent bond.

[0285] "Halogen" refers to fluorine, chlorine, bromine, or iodine.

[0286] "Halogenated" means that one or more (such as 1, 2, 3, 4 or 5) hydrogen atoms in a group are replaced by halogens.

[0287] For example, "halogenated alkyl" refers to an alkyl group in which the hydrogen atom is replaced by one or more (e.g., 1, 2, 3, 4, or 5) halogens, wherein the definition of alkyl is as described above. Preferably, it is a halogenated C. 1-8 Alkyl groups, preferably halogenated C 1-6 Alkyl, more preferably halogenated C 1-3 Alkyl groups. Examples of haloalkyl groups include (but are not limited to) monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, monobromoethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, etc.

[0288] For example, "haloalkoxy" refers to an alkoxy group where the hydrogen atom is replaced by one or more (e.g., 1, 2, 3, 4, or 5) halogens, as defined above. Preferably, it is a halogenated C. 1-8 Alkoxy, more preferably halogenated C 1-6 Alkoxy, more preferably halogenated C 1-3 Alkyl groups. Halogenated alkoxy groups include (but are not limited to) trifluoromethoxy, trifluoroethoxy, monofluoromethoxy, monofluoroethoxy, difluoromethoxy, difluoroethoxy, etc.

[0289] For example, "halogenated cycloalkyl" refers to a cycloalkyl group in which the hydrogen atoms are replaced by one or more (e.g., 1, 2, 3, 4, or 5) halogens, as defined above. Preferably, the halogenated C atoms are used. 3-8 Cycloalkyl, more preferably halogenated C 3-6 Cycloalkyl groups. Halogenated cycloalkyl groups include (but are not limited to) trifluorocyclopropyl, monofluorocyclopropyl, monofluorocyclohexyl, difluorocyclopropyl, difluorocyclohexyl, etc.

[0290] "Deuterated alkyl" refers to an alkyl group in which one or more (e.g., 1, 2, 3, 4, or 5) deuterium atoms are substituted for hydrogen atoms, as defined above. Preferably, it is a deuterated C-type alkyl group. 1-8 Alkyl groups, more preferably deuterated C4 groups 1-6 Alkyl groups, more preferably deuterated C4 groups 1-3 Alkyl groups. Examples of deuterated alkyl groups include (but are not limited to) monodeuterated methyl, monodeuterated ethyl, dideuterated methyl, dideuterated ethyl, trideuterated methyl, trideuterated ethyl, etc.

[0291] "Amino" refers to NH2, "cyano" refers to CN, "nitro" refers to NO2, "benzyl" refers to -CH2-phenyl, "oxo" refers to =O, "carboxyl" refers to -C(O)OH, "acetyl" refers to -C(O)CH3, "hydroxymethyl" refers to -CH2OH, "hydroxyethyl" refers to -CH2CH2OH or -CHOHCH3, "hydroxyl" refers to -OH, "mercapto" refers to SH, and the structure of "cyclopropyl" is:

[0292] "Substituted" refers to one or more hydrogen atoms in a group, preferably 1 to 5 hydrogen atoms independently substituted by a corresponding number of substituents, more preferably 1 to 3 hydrogen atoms independently substituted by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (through experiment or theory) possible or impossible substitutions without much effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom having an unsaturated bond (such as an alkene).

[0293] Unless otherwise defined, the phrase "substituents selected independently of each other" in this invention means that when one or more hydrogen atoms on a group are replaced by substituents, the types of substituents may be the same or different, and the selected substituents are of independent types.

[0294] Unless otherwise defined, the phrase "...same or different, and each independently being..." in this invention means that when there are more than one identical substituent group in the general formula, the group can be the same or different, and each can be an independent type. For example, L is (CR 01 R 02 ) s When s is 2, that is, L is (CR) 01 R 02 )-(CR 01 R 02 ), where the two R 01 The two Rs can be the same or different. 02 They can be the same or different, and can be independent types. For example, L can be C(CH3)(CN)-C(CH2CH3)(OH), C(CH3)(CN)-C(CH3)(OH) or C(CN)(CH2CH3)-C(OH)(CH2CH3).

[0295] Unless otherwise defined, any group herein may be substituted or unsubstituted. When the above groups are substituted, the substituents are preferably 1 to 5 groups, independently selected from cyano, halogen (preferably fluorine or chlorine), C 1-8 Alkyl (preferably C) 1-6 Alkyl, more preferably C 1-3 Alkyl), C 1-8 Alkoxy (preferably C)1-6 Alkoxy, more preferably C 1-3 alkoxy), halogenated C 1-8 Alkyl (preferably halogenated C) 1-6 Alkyl, more preferably halogenated C 1-3 Alkyl), C 3-8 cycloalkyl (preferably C) 3-6 cycloalkyl), halogenated C 1-8 Alkoxy (preferably halogenated C) 1-6 Alkoxy, more preferably halogenated C 1-3 alkoxy), C 1-8 Alkyl-substituted amino groups, halogenated C 1-8 Alkyl-substituted amino, acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxyl, nitro, C 6-10 Aryl (preferably phenyl), C 3-8 Cycloalkyloxy (preferably C) 3-6 cycloalkyloxy), C 2-8 alkenyl (preferably C) 2-6 alkenyl, more preferably C 2-4 alkenyl), C 2-8 alkynyl group (preferably C) 2-6 Alkyne group, more preferably C 2-4 alkynyl group), -CONR a0 R b0 -C(O)OC 1-10 Alkyl (preferably -C(O)OC) 1-6 Alkyl groups, more preferably -C(O)OC 1-3 Alkyl groups, -CHO, -OC(O)C 1-10 Alkyl groups (preferably -OC(O)C) 1-6 Alkyl groups, more preferably -OC(O)C 1-3 alkyl), -SO2C 1-10 Alkyl groups (preferably -SO2C) 1-6 Alkyl groups, more preferably -SO2C 1-3 alkyl), -SO2C 6-10 Aryl (preferably -SO2C6 aryl, such as -SO2-phenyl), -COC 6-10 aryl (preferably -COC6 aryl, such as -CO-phenyl), 4 to 6 membered saturated or unsaturated mono-heterocyclic rings, 4 to 6 membered saturated or unsaturated mono-cyclic rings, 5 to 6 membered monocyclic heteroaryl rings, 8 to 10 membered bicyclic heteroaryl rings, spirocyclic rings, spiroheterocyclic rings, bridged rings or bridged heterocyclic rings, wherein R a0 R b0 Each is independently hydrogen or C 1-3 alkyl.

[0296] In this invention, when two or more "preferred" options appear in a solution, any two "preferred" options can be independent of each other.

[0297] In this invention, when the number of substituents is greater than 1, any two substituents can be the same or different. For example, they can be substituted with two identical or different halogens, or with one halogen and one hydroxyl group.

[0298] The various substituents described above can themselves be replaced by the groups described in this article.

[0299] Pharmaceutical Composition

[0300] Typically, the compounds of this invention, or their pharmaceutically acceptable salts or stereoisomers, can be formulated with one or more pharmaceutical carriers into suitable dosage forms for administration. These dosage forms are suitable for oral, rectal, topical, intraoral, and other non-gastrointestinal administration (e.g., subcutaneous, intramuscular, intravenous, etc.). For example, suitable dosage forms for oral administration include capsules, tablets, granules, and syrups. The compounds of this invention contained in these formulations can be solid powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; water-in-oil or oil-in-water emulsions, etc. The above dosage forms can be prepared from the active compound and one or more carriers or excipients using conventional pharmaceutical methods. The carriers must be compatible with the active compound or other excipients. The active compound can form a solution or suspension with the carrier.

[0301] "Pharmaceutically acceptable carrier" refers to a non-toxic, inert, solid, or semi-solid substance or liquid filling machine, diluent, encapsulation material, or excipient or any type of excipient that is compatible with patients, preferably mammalian, more preferably human, and suitable for delivering an active agent to a target site without terminating the agent's activity.

[0302] "The compound of the present invention" refers to the compound of formula (I) of the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, which has the effect of regulating the level of proteins such as GSPT1 and inhibiting the proliferation of tumor cells, as well as intermediate compounds for preparing the compound of formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0303] The compositions of the present invention are formulated, quantified, and administered in accordance with medical practice guidelines. The "therapeutic effective amount" of the compound administered is determined by factors such as the specific condition to be treated, the individual being treated, the cause of the condition, the target of the drug, and the route of administration.

[0304] "Therapeutic effective amount" refers to the amount of the compound of the present invention that will elicit a biological or medical response in an individual, such as reducing or inhibiting enzyme or protein activity or improving symptoms, alleviating symptoms, slowing or delaying disease progression, or preventing disease.

[0305] "Patient" refers to an animal, preferably a mammal, and more preferably a human. The term "mammal" refers to warm-blooded vertebrate mammals, including animals such as cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, rats, pigs, and humans.

[0306] "Treatment" refers to reducing, slowing the progression of, attenuating, preventing, or maintaining an existing disease or condition (such as cancer). Treatment also includes curing, preventing the development of, or reducing to some extent one or more symptoms of a disease or condition.

[0307] The term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. Pharmaceutically acceptable acid addition salts are salts formed with inorganic or organic acids that retain the bioavailability of the free base without other side effects. These salts can be prepared using methods known in the art.

[0308] "Pharmaceutically acceptable base addition salts" include, but are not limited to, salts of inorganic bases such as sodium, potassium, calcium, and magnesium salts. They also include, but are not limited to, salts of organic bases such as ammonium, triethylamine, lysine, and arginine salts. These salts can be prepared using methods known in this field.

[0309] When the compound represented by formula (I) contains one or more chiral centers, it can exist in different optically active forms. When the compound of formula (I) contains one chiral center, the compound comprises a pair of enantiomers. The two enantiomers of the compound, as well as mixtures of the pair of enantiomers, such as racemic mixtures, are also within the scope of protection of this invention. Enantiomers can be resolved by methods known in the art, such as crystallization and chiral chromatography. When the compound of formula (I) contains more than one chiral center, the compound comprises enantiomers and diastereomers. All enantiomers and diastereomers of the compound, as well as mixtures of enantiomers, mixtures of diastereomers, and mixtures of enantiomers and diastereomers, are also within the scope of protection of this invention. Enantiomers and diastereomers can be resolved by methods known in the art, such as crystallization and preparative chromatography.

[0310] As used in this article, the waveform line refers to... The asterisk (*) indicates "".

[0311] The term "drug-to-antibody ratio" or "DAR" refers to the amount of drug, such as the amount of small molecule toxin conjugated to an antibody-drug conjugate (ADC). The DAR value of an ADC can range from 1 to 15, but depends on the number of binding sites on the antibody; higher loadings (e.g., 18) are also possible. The term DAR may be used when referring to the amount of drug loaded onto a single antibody, or alternatively, when referring to the average or mean DAR of a group of ADCs. In some embodiments, the ADC of the present invention has a DAR value of any value between 1 and 10, such as 1.97, 2, 3.43, 4, and 4.75. The ADC is generated by coupling after reduction of an interchain disulfide.

[0312] In this invention, the terms "conjugate," "antibody-drug conjugate," and "ADC" are used interchangeably and all refer to substances obtained by linking a bioactive compound fragment (such as a drug molecule represented by formula (I) or (II)) with an antibody or its antigen-binding fragment portion, having a structure as shown in formula (B). The bioactive compound fragment and the target portion (the antibody or its antigen-binding fragment portion) are linked by a linker. The linker can cleave under specific environments (such as intracellular low pH environments) or under specific actions (such as lysosomal protease action), causing the bioactive compound fragment to separate from the target portion. In some embodiments, the linker comprises a linker group with a functional group capable of reacting with a thiol group and a linker group of a non-cleavable or cleavable unit, such as a peptide or a disulfide bond.

[0313] In this invention, the term "drug" refers to the compound represented by formula (I) or formula (II) of this application, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, which has significant protein regulatory effects and enhanced cell proliferation inhibition effects, and can be used as a GSPT1 degrading agent.

[0314] In this invention, the term "antibody" refers to an antibody against Her 2, such as pertuzumab or trastuzumab.

[0315] In this invention, the term "linker-drug" refers to the compound represented by formula (A), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, which is composed of a linker group T, a linker group L, and a drug connected by covalent bonds. The linker group T is a component of the antibody-drug conjugate or linker, and its function is to link the antibody or its antigen-binding fragment that binds to the target site to the remaining portion of the antibody-drug conjugate. The linker group T contains a functional group capable of reacting with a thiol group and a structural fragment for binding the linker group to a short peptide composed of amino acid residues, such as...

[0316] In this invention, the linker group L is a component of a drug conjugate or linker, which is used to bind the linker group to the structural fragment of the drug. It contains structural fragments that can be broken under specific environments or specific effects, such as short peptides composed of 2 to 4 amino acid residues, wherein the amino acid residues are selected from natural amino acid residues and non-natural amino acid residues.

[0317] Preparation method

[0318] This invention provides a method for preparing compounds of formula (I), which can be synthesized using standard synthetic techniques known to those skilled in the art or by combining methods known in the art with the methods described herein. The solvents, temperatures, and other reaction conditions given in this invention can be varied according to the art. The reactions can be used sequentially to provide the compounds of this invention, or they can be used to synthesize fragments subsequently added by the methods described herein and / or methods known in the art.

[0319] The compounds described in this invention can be synthesized using appropriate, selectable starting materials, similar to those described in the examples below or relevant publications available to those skilled in the art. The starting materials used to synthesize the compounds described in this invention can be synthesized or are available from commercial sources. The compounds described in this invention and other related compounds with different substituents can be synthesized using techniques and starting materials known to those skilled in the art. General methods for preparing the compounds of this invention can be derived from reactions known in the art, and these reactions can be modified by reagents and conditions deemed appropriate by those skilled in the art to introduce various moieties in the molecules provided by this invention.

[0320] As an example, the general synthetic route for the compound of this application (IA) is shown in Scheme 1 below:

[0321] Option 1

[0322] Compound (IA-1) is reacted with compound (IA-2) or a salt thereof in the presence of triphosgene to generate compound (IAa). The reaction can be carried out in an inert solvent at a specific temperature (e.g., -20°C to 80°C, preferably 0°C to 60°C, more preferably 20°C to 60°C), by reacting compound (IA-1) with triphosgene for a period of time (e.g., 5 minutes to 48 hours, preferably 0.1 hours to 5 hours). The resulting mixture is then reacted with compound (IA-2) or a salt thereof in the presence of a basic reagent at a specific temperature (e.g., -20°C to 80°C, preferably 0°C to 60°C, more preferably 20°C to 60°C) for a period of time (e.g., 5 minutes to 48 hours, preferably 0.1 hours to 5 hours) to obtain compound (IAa). The inert solvent and basic reagent can be those known in the art, wherein the inert solvent can be selected from, for example, C... 1-4 Alkyl alcohols, toluene, xylene, methyl tert-butyl ether, tetrahydrofuran, 1,4-dioxane, diethyl ether, dichloromethane, chloroform, 1,2-dichloroethane, ethyl acetate, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, etc., or combinations thereof. Alkaline reagents may be selected from ammonia, triethylamine, diisopropylethylamine, N,N-dimethylaniline, tetramethylethylenediamine, tributylamine, sodium methoxide, sodium ethoxide, potassium ethoxide, sodium tert-butoxide, potassium tert-butoxide, lithium tert-butoxide, 1,8-diazabicyclo[5,4,0]-7-undecene (DBU), pyrazole, imididine, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, sodium acetate, sodium phenolate, sodium benzoate, sodium citrate, N-methylmorpholine, pyridine, or combinations thereof. Wherein R in formula (IA-1) p The nitrogen protecting group can be a commonly known nitrogen protecting group in the art, such as benzyloxycarbonyl (-Cbz) or tert-butyloxycarbonyl (-Boc).

[0323] Compound of formula (IAa) is obtained by removing the nitrogen protecting group from the compound under suitable conditions, such as acidic conditions or palladium-carbon / hydrogen reduction conditions.

[0324] As an example, the general synthetic route for the compound of this application (IA) is shown in Scheme 2 below:

[0325] Option 2

[0326] Compound (IA-3) is reacted with compound (IA-2) or a salt thereof in the presence of triphosgene to generate compound (IAb). The reaction can be carried out in an inert solvent at a specific temperature (e.g., -20°C to 80°C, preferably 0°C to 60°C, more preferably 20°C to 60°C), by reacting compound (IA-3) with triphosgene for a period of time (e.g., 5 minutes to 48 hours, preferably 0.1 hours to 5 hours). The resulting mixture is then reacted with compound (IA-2) or a salt thereof in the presence of a basic reagent at a specific temperature (e.g., -20°C to 80°C, preferably 0°C to 60°C, more preferably 20°C to 60°C) for a period of time (e.g., 5 minutes to 48 hours, preferably 0.1 hours to 5 hours) to obtain compound (IAb). The inert solvent and basic reagent can be those known in the art, wherein the inert solvent can be selected from, for example, C... 1-4 Alkyl alcohols, toluene, xylene, methyl tert-butyl ether, tetrahydrofuran, 1,4-dioxane, diethyl ether, dichloromethane, chloroform, 1,2-dichloroethane, ethyl acetate, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, etc., or combinations thereof. Alkaline reagents may be selected from ammonia, triethylamine, diisopropylethylamine, N,N-dimethylaniline, tetramethylethylenediamine, tributylamine, sodium methoxide, sodium ethoxide, potassium ethoxide, sodium tert-butoxide, potassium tert-butoxide, lithium tert-butoxide, 1,8-diazabicyclo[5,4,0]-7-undecene (DBU), pyrazole, imididine, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, sodium acetate, sodium phenolate, sodium benzoate, sodium citrate, N-methylmorpholine, pyridine, or combinations thereof. Wherein R in formula (IA-3) p It represents a nitrogen protecting group, such as commonly known nitrogen protecting groups in the art, such as -Cbz or -Boc.

[0327] Compound of formula (IAb) is denitrified under suitable conditions, such as acidic conditions or palladium-carbon / hydrogen reduction conditions, to obtain compound of formula (IAc).

[0328] Compound of formula (IAc) is obtained by reductive amination of the compound and the corresponding aldehyde or ketone compound. The reductive amination reaction is carried out in an inert solvent at a specific temperature (e.g., -20°C to 80°C, preferably 0°C to 60°C, more preferably 20°C to 60°C), by reacting the compound of formula (IAc), the aldehyde or ketone compound with a reducing agent for a period of time (e.g., 0.5 hours to 48 hours, preferably 0.5 hours to 5 hours) to obtain the compound of formula (IA). The inert solvent and reducing agent can be those known in the art, and the reducing agent can be selected from, for example, tetrabutylamine borohydride, sodium malonyloxyborohydride, sodium triacetoxyborohydride, sodium cyanoborohydride, sodium borohydride, lithium borohydride, potassium borohydride, borane, etc. The inert solvent can be selected from, for example, C 1-4 Alkyl alcohols, toluene, xylene, methyl tert-butyl ether, tetrahydrofuran, 1,4-dioxane, diethyl ether, dichloromethane, chloroform, 1,2-dichloroethane, ethyl acetate, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, etc., or combinations thereof.

[0329] As an example, the compound of this application (IB) when R 62 The general synthetic route for compounds of formula (IBb) with hydrogen is shown in Scheme 3 below:

[0330] Option 3

[0331] The compound of formula (IB-1) is reacted with the compound of formula (IA-2) or a salt thereof in the presence of triphosgene to generate the compound of formula (IBa). The reaction can be carried out in an inert solvent at a specific temperature (e.g., -20°C to 80°C, preferably 0°C to 60°C, more preferably 20°C to 60°C), by reacting the compound of formula (IB-1) with triphosgene for a period of time (e.g., 5 minutes to 48 hours, preferably 0.1 hours to 5 hours). The resulting mixture is then reacted with the compound of formula (IA-2) or a salt thereof in the presence of a basic reagent at a specific temperature (e.g., -20°C to 80°C, preferably 0°C to 60°C, more preferably 20°C to 60°C) for a period of time (e.g., 5 minutes to 48 hours, preferably 0.1 hours to 5 hours) to obtain the compound of formula (IBa). The inert solvent and basic reagent can be those known in the art, wherein the inert solvent can be selected from, for example, C... 1-4Alkyl alcohols, toluene, xylene, methyl tert-butyl ether, tetrahydrofuran, 1,4-dioxane, diethyl ether, dichloromethane, chloroform, 1,2-dichloroethane, ethyl acetate, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, etc., or combinations thereof. Alkaline reagents may be selected from ammonia, triethylamine, diisopropylethylamine, N,N-dimethylaniline, tetramethylethylenediamine, tributylamine, sodium methoxide, sodium ethoxide, potassium ethoxide, sodium tert-butoxide, potassium tert-butoxide, lithium tert-butoxide, 1,8-diazabicyclo[5,4,0]-7-undecene (DBU), pyrazole, imididine, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, sodium acetate, sodium phenolate, sodium benzoate, sodium citrate, N-methylmorpholine, pyridine, or combinations thereof. Wherein R in formula (IB-1) p It represents a nitrogen protecting group, such as commonly known nitrogen protecting groups in the art, such as -Cbz or -Boc.

[0332] Compound of formula (IBa) is denitrified under suitable conditions, such as acidic conditions or palladium-carbon / hydrogen reduction conditions, to obtain compound of formula (IBb).

[0333] The main advantages of this invention include:

[0334] Compared with existing compounds, the isoindoline compound containing a phenylethoxyalkyleneamine structure of this application, as a protein degrader such as GSPT1, has a significantly enhanced inhibitory effect on the proliferation of tumor cells, such as human breast ductal carcinoma cells, and its inhibitory activity against the proliferation of MDA-MB-453 cells is IC50. 50 Values ​​less than 500 nmol / L (nM), and in some implementations, IC50 values ​​less than 300 nM. 50 Values, in some implementations, have ICs with a value less than 100 nM. 50 Values, in some implementations, have ICs with a value less than 10 nM. 50 The value is at least twice that of existing compounds. Therefore, it can be used to prepare protein degrading agents such as GSPT1 for the treatment or prevention of diseases related to or caused by GSPT1.

[0335] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions as described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Percentages and parts are by weight unless otherwise stated. Unless otherwise defined, the terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be used in the present invention.

[0336] The known starting materials can be synthesized using methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, and Darui Chemicals. Unless otherwise specified, the reactions in the examples were carried out under a nitrogen or argon atmosphere. DCM: Dichloromethane, DMF: Dimethylformamide, THF: Tetrahydrofuran, Pd(dppf)Cl2: [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride, Pin2B2: Pinacol diboronate, SEM-Cl: 2-(trimethylsilyl)ethoxymethyl chloride, ACN: Acetonitrile, n-BuLi: n-Butyllithium, KOAc: Potassium acetate, HATU: 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, EDCl: 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, DIPEA: N,N-diisopropylethylamine, EEDQ: 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline, MeOH: Methanol, EA: Ethyl acetate, PE: Petroleum ether, FA: Formic acid, TBAF: Tetrabutylammonium fluoride.

[0337] Unless otherwise specified, the percentage contents mentioned in this invention refer to mass percentage for solid-liquid mixtures and solid-phase-solid mixtures, and volume percentage for liquid-phase-liquid mixtures. The final product of this invention can be obtained through preparation and purification. The purification and separation conditions are determined experimentally by those skilled in the art based on the characteristics of the compounds and conventional knowledge in the field.

[0338] As used in this article, room temperature refers to approximately 20-30°C.

[0339] Preparation of intermediate compound V1

[0340] Step 1: Add V1-1 (50.0 g, 231.92 mmol) to a 2 L three-necked reaction flask and stir in an ice-water bath at 0 °C under nitrogen protection. Slowly add boranetetrahydrofuran (1 M, 510.23 mmol, 510.2 mL) dropwise. After the addition is complete, continue stirring for 12 hours. After the reaction cools to room temperature, place it in a water bath and stir. Slowly add methanol dropwise to quench the reaction until no more bubbles are produced. Then, evaporate to dryness under reduced pressure. The crude product is purified by Combi Flash silica gel column chromatography (120 g, 0–30 vol% ethyl acetate / petroleum ether) to obtain orange-red solid V1-2 (45.46 g, 97.23% yield). 1 H NMR (400MHz, CDCl3) δ8.25 (d, J = 2.4Hz, 1H), 8.07 (dd, J = 8.5, 2.4Hz, 1H), 7.49 (d, J = 8.4Hz, 1H), 3.94 (t, J = 6.5Hz, 2H), 3.11 (t, J = 6.5Hz, 2H). LC-MS m / z(ESI):201.9[M+1] + .

[0341] Step 2: Dissolve V1-2 (10.09 g, 50.05 mmol) in toluene (100.0 mL), add tetrabutylammonium bisulfate (10.03 g, 29.53 mmol), and stir in an ice-water bath at 0 °C. Slowly add tert-butyl bromoacetate (39.05 g, 200.19 mmol), followed by dropwise addition of sodium hydroxide (9.61 g, 240.23 mmol) dissolved in H2O (60.0 mL). After the addition is complete, gradually raise the temperature to room temperature and continue stirring for 12 hours. Pour the reaction mixture into a separatory funnel, separate the organic phase, extract the aqueous phase with ethyl acetate (60 mL), combine the organic phases, dry with anhydrous sodium sulfate, filter, and evaporate to dryness under reduced pressure. The crude product is then evaporated to dryness under reduced pressure using an oil pump to obtain a yellow oily liquid V1-3 (15.6 g, 98.72% yield). 1 H NMR (400MHz, CDCl3) δ8.18 (d, J=2.4Hz, 1H), 8.03 (dd, J=8.5, 2.4Hz, 1H), 7.57 (d, J= 8.5Hz, 1H), 3.92 (s, 2H), 3.78 (t, J = 6.5Hz, 2H), 3.12 (t, J = 6.5Hz, 2H), 1.43 (s, 9H).

[0342] Step 3: V1-3 (15.6 g, 49.41 mmol) was dissolved in dichloromethane (44.0 mL), stirred at 20 °C, and trifluoroacetic acid (28.17 g, 247.03 mmol) was added. The reaction mixture was stirred for another 24 hours. The reaction solution was evaporated to dryness under reduced pressure. The crude product was then dried under reduced pressure using an oil pump, and purified by Combi Flash silica gel column chromatography (120 g, 0–38 vol% ethyl acetate / petroleum ether) to obtain a yellow oily liquid V1 (10.24 g, 79.83% yield). 1 H NMR (400MHz, CDCl3) δ8.25 (d, J = 2.3 Hz, 1H), 8.07 (dd, J = 8.5, 2.3 Hz, 1H), 7.53 (d, J = 8.5 Hz, 1H), 4.12 (s, 2H), 3.86 (t, J = 6.5 Hz, 2H), 3.17 (t, J = 6.5 Hz, 2H). LC-MS m / z(ESI):260.1[M+1] + ;282.0[M+23] + .

[0343] Preparation of intermediate compound V2

[0344] Step 1: V2-1 (413 mg, 1 mmol) was dissolved in N,N-dimethylformamide (10 mL), and tris(dibenzylacetone)palladium (47 mg, 0.05 mmol), 1,1'-bis(diphenylphosphine)ferrocene (28 mg, 0.05 mmol), zinc acetate (91 mg, 0.5 mmol), and zinc cyanide (170 mg, 1.5 mmol) were added. The mixture was reacted under nitrogen protection at 120 °C for 2 hours. The solvent was removed by concentration (i.e., concentrated to dryness), and the product was separated by silica gel column chromatography (dichloromethane:methanol = 100:1 to 9:1) to directly obtain brown solid V2-2 (390 mg, crude product). LC-MS m / z (ESI): 288.0 [M+1] + .

[0345] Step 2: V2-2 (390 mg, 1.3 mmol) was dissolved in methanol (10 mL), hydrochloric acid (1 mL), and platinum dioxide (150 mg, 0.6 mmol) was added. The reaction was carried out under hydrogen atmosphere for 15 h. The mixture was then separated by silica gel column chromatography (dichloromethane:methanol = 100:1 to 2:1) to directly give a grayish-white solid V2 (360 mg, 91.1% yield). LC-MS m / z (ESI): 292.0 [M+1] + .

[0346] Preparation of intermediate compound V3

[0347] Step 1: V1 (10.65 g, 41.02 mmol) and methylamine (3.32 g, 49.22 mmol, hydrochloride) were dissolved in N,N-dimethylformamide (50.62 mL), and HATU (18.55 g, 49.22 mmol) and diisopropylethylamine (15.89 g, 123.05 mmol, 21.41 mL) were added sequentially. The mixture was stirred at 20 °C for 4 hours. The reaction mixture was poured into a separatory funnel, and water (300 mL) was added. The mixture was extracted with ethyl acetate (60 mL × 2). The organic phases were combined and washed with saturated brine (80 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness under reduced pressure. The crude product was purified by Combi Flash silica gel column chromatography (120 g, 0–65% ethyl acetate / petroleum ether) to obtain a yellow oily liquid V3-1 (10.41 g, 93.07% yield). LC-MS m / z(ESI): 273.1 [M+1] + .

[0348] Step 2: Add V3-1 (10.41 g, 38.18 mmol) to a 500 mL three-necked reaction flask and dissolve it in tetrahydrofuran (60 mL). Under nitrogen protection, place the flask in an ice-water bath at 0 °C and stir. Slowly add borane tetrahydrofuran (1 M, 381.76 mmol, 382 mL). After the addition is complete, place the reaction flask in a 70 °C oil bath and heat and stir for 12 hours. After the reaction cools to room temperature, place the flask in a water bath and stir. Slowly add methanol to quench the reaction mixture until no more bubbles are produced. Then, evaporate the mixture to dryness under reduced pressure. Add a small amount of methanol to the resulting liquid to dissolve it and stir. Add hydrochloric acid / dioxane (4 M, 191 mmol, 47.7 mL) and stir for ten minutes. LC-MS analysis showed that the intermediate had been completely converted to the target product. The solution was evaporated to dryness under reduced pressure, dissolved in ethyl acetate (100 mL), and poured into a separatory funnel. The pH of the aqueous solution was adjusted to weakly alkaline by adding saturated sodium bicarbonate (200 mL). Extraction was performed with ethyl acetate (80 mL × 2). The organic phases were combined and washed with saturated brine (100 mL). The solution was dried over anhydrous sodium sulfate. The mixture was filtered and evaporated to dryness under reduced pressure. The crude product was then dried under reduced pressure using an oil pump to give a yellow oily liquid, V3-2 (9.81 g, 79.46% yield). LC-MS m / z (ESI): 259.1 [M+1] + .

[0349] Step 3: V3-2 (9.81 g, 30.34 mmol) and di-tert-butyl dicarbonate (7.95 g, 36.40 mmol) were dissolved in tetrahydrofuran (60.0 mL), stirred at 20 °C, and triethylamine (9.21 g, 91.01 mmol, 12.69 mL) was slowly added. After the addition was complete, the reaction mixture was stirred for another 12 hours. The reaction solution was evaporated to dryness under reduced pressure. The crude product was purified by Combi Flash silica gel column chromatography (120 g, 0–16% ethyl acetate / petroleum ether) to obtain a yellow oily liquid V3-3 (5.63 g, 51.72% yield). LC-MS m / z (ESI): 259.1 [M+1-100] + .

[0350] Step 4: V3-3 (5.63 g, 15.69 mmol) and ammonium chloride (2.52 g, 47.07 mmol) were dissolved in ethanol (100 mL) and water (20 mL). Reduced iron powder (4.38 g, 78.45 mmol) was added. After the addition was complete, nitrogen gas was introduced, and the mixture was heated and stirred in an oil bath at 80 °C for 4 hours. The cooled reaction solution was filtered through a diatomaceous earth layer, and the filter cake was washed with ethanol (80 mL × 3). The filtrates were combined and evaporated to dryness under reduced pressure. The crude product was purified by Combi Flash silica gel column chromatography (40 g, 0–28% ethyl acetate / petroleum ether) to obtain a yellow oily liquid V3 (4.71 g, 91.29% yield). LC-MS m / z (ESI): 229.1 [M+1-100] + .

[0351] Preparation of intermediate compound V4

[0352] Step 1: V1 (2.59 g, 10 mmol) was dissolved in N,N-dimethylformamide (20 mL), and trifluoroethylamine (1 g, 10 mmol), HATU (5.7 g, 1.5 mmol), and triethylamine (2 g, 20 mmol) were added. The mixture was reacted at room temperature for 1 hour, then extracted with water and ethyl acetate, and concentrated to dryness to obtain a brown solid, V4-1 (3.4 g, crude product). LC-MS m / z (ESI): 341.0 [M+1] + .

[0353] Step 2: V4-1 (3.4 g, 10 mmol) was dissolved in tetrahydrofuran (15 mL), and boranetetrahydrofuran (60 mL) was added. The reaction was carried out at 70 °C for 2 hours, quenched with methanol, and concentrated to dryness to obtain a yellow oily substance, V4-2 (3.9 g, crude product). LC-MS m / z (ESI): 327.0 [M+1] + .

[0354] Step 3: V4-2 (3.9 g, 12 mmol) was dissolved in tetrahydrofuran (40 mL), and di-tert-butyl dicarbonate (2.5 g, 12 mmol) and triethylamine (2.5 g, 25 mmol) were added. The mixture was reacted at room temperature for 12 hours, concentrated to dryness, and separated by silica gel column chromatography (ethyl acetate / petroleum ether = 31%) to give a yellow oil, V4-3 (2.9 g, 56.9% yield). LC-MS m / z (ESI): 327.0 [M+1-100] + .

[0355] Step 4: V4-3 (2.9 g, 6.8 mmol) was dissolved in ethanol (40 mL) and water (20 mL). Reduced iron powder (2.1 g, 40 mmol) and ammonium chloride (2.1 g, 40 mmol) were added. The mixture was reacted at 86 °C for 2 hours, filtered, and the filtrate was extracted and concentrated to dryness to obtain a yellow oily substance, V4 (2 g, crude product). LC-MS m / z (ESI): 297.0 [M+1-100] + .

[0356] Preparation of intermediate compound 3k

[0357] Step 1: Compound 3a (10 g, 56.13 mmol) was dissolved in anhydrous dichloromethane (80 mL), and 2,2-dimethyl-1,3-dioxane-4,6-dione (7.69 g, 53.33 mmol) was added. After the reaction temperature was lowered to 0 °C, 4-dimethylaminopyridine (3.43 g, 28.07 mmol) and triethylamine (13.06 g, 129.11 mmol) were added. Finally, solid EDCl (16.14 g, 84.20 mmol) was slowly added. After reacting at 0 °C for half an hour, the temperature was gradually increased to room temperature, and then the reaction was carried out for 4 hours. LCMS monitoring showed that the starting material had almost disappeared, and a new major peak was obtained as an intermediate state (LCMS did not determine the molecular weight). After concentration, a brown oily crude product 3b was obtained, which was directly used in the next reaction step.

[0358] Step 2: The crude product 3b, after being evaporated to dryness, was added to toluene (250 mL) and concentrated hydrochloric acid (12 M, 35 mL). The reaction was gradually heated to 65 °C in an oil bath for 12 hours. The reaction solution was carefully neutralized to pH approximately 9 with saturated sodium carbonate aqueous solution, and then extracted three times with ethyl acetate. The combined organic phases were dried over sodium sulfate, washed with sodium bicarbonate aqueous solution, evaporated to dryness under reduced pressure, and concentrated to obtain 3c (7.8 g, 78.8% yield) as a white solid. LC-MS m / z (ESI): 177.1 [M+1] + .

[0359] Step 3: Compound 3c (3.6 g, 20.43 mmol) was dissolved in anhydrous methanol (100 mL), and sodium acetate (13.41 g, 163.48 mmol) and hydroxylamine hydrochloride (11.36 g, 163.48 mmol) were added. The mixture was heated in an oil bath to 45 °C and stirred for 2 hours. The methanol was removed under reduced pressure, and ethyl acetate and water were added. The organic phase was extracted twice with ethyl acetate, and the two organic phases were combined, dried over sodium sulfate, and concentrated to give 3d (3.6 g, 92.1% yield) as a white solid. LC-MS m / z (ESI): 192.1 [M+1] + .

[0360] Step 4: Compound 3d (4.5 g, 23.54 mmol) was dissolved in anhydrous methanol (100 mL), and wet palladium-carbon (1.09 g, 10% wt) was added. After three purgings with hydrogen balloons, the reaction mixture was heated to 40 °C and reacted for 3 hours. After cooling, the mixture was filtered through diatomaceous earth. The filter cake was washed with methanol. Product 3e (3.5 g, crude product) was used directly as the filtrate for the next reaction without rotary evaporation. LC-MS m / z (ESI): 178.1 [M+1] + .

[0361] Step 5: Compound 3e (3.5 g, 19.75 mmol) was dissolved in anhydrous methanol (120 mL) (this reaction system is the filtrate after filtration in the previous step), and triethylamine (6.38 g, 49.38 mmol, 8.60 mL) and di-tert-butyl dicarbonate (6.47 g, 29.63 mmol, 6.81 mL) were added. The reaction mixture was stirred at room temperature for 1 hour. The reaction solution was evaporated to dryness under reduced pressure to obtain a crude brown oil. The brown oil was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 3f (3.6 g, 65.7% yield) as a brown oil; LC-MS m / z (ESI): 222.1 [M+1-56] + .

[0362] Step 6: Compound 3f (3.6 g, 12.98 mmol) was dissolved in tetrahydrofuran (15 mL), and then NaOH (778.90 mg, 19.47 mmol) dissolved in water (10 mL) was slowly added dropwise. The reaction mixture was stirred at room temperature for 1.5 hours. The reaction solution was neutralized to pH 5 with 2 M hydrochloric acid, and then the organic phase was extracted with ethyl acetate. The combined organic phases were evaporated under reduced pressure to give 3 g (3.5 g, 91.3% yield) of compound as a white solid; LC-MS m / z (ESI): 317.8 [M+23] + .

[0363] Step 7: Dissolve 3 g (1 g, 3.39 mmol) of compound in anhydrous methanol (6 mL) and ethyl acetate (6 mL). After lowering the reaction temperature to -20 °C, trimethylsilyldiazomethane (2 M, 8.47 mL) was slowly added dropwise under argon protection. The reaction mixture was stirred for 1 hour at -10 °C to room temperature. The reaction solution was carefully quenched with water, and then extracted with ethyl acetate in the aqueous phase. The organic phases were combined and evaporated to dryness under reduced pressure to obtain compound 3h (1.1 g, crude product) as a brown oil. The product was used directly in the next reaction. LC-MS m / z (ESI): 332.2 [M+23] + .

[0364] Step 8: Compound 3h (0.85 g, 2.75 mmol) was dissolved in anhydrous tetrahydrofuran (15 mL). The reaction temperature was lowered to 0 °C, and triphenylphosphine (1.08 g, 4.12 mmol) and carbon tetrabromide (1.37 g, 4.12 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. The reaction solution was evaporated to dryness under reduced pressure to obtain a crude brown oil. The brown oil was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to give compound 3i (166 mg, 16.2% yield) as a brown oil; LC-MS m / z (ESI): 316.1, 318.1 [M+1-56] + .

[0365] Step 9: Compound 3i (160 mg, 429.82 μmol) was dissolved in anhydrous acetonitrile (7 mL), and N,N-diisopropylethylamine (277.75 mg, 2.15 mmol) was added. The reaction mixture was heated to 85 °C and stirred for 16 hours. The reaction solution was evaporated to dryness under reduced pressure to obtain a crude brown oil. The brown oil was subjected to silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain compound 3j (145 mg, 87.1% yield) as a brown oil; LC-MS m / z (ESI): 388.2 [M+1] + .

[0366] Step 10: Compound 3j (160 mg, 412.98 μmol) was dissolved in anhydrous dichloromethane (3 mL), and trifluoroacetic acid (706.34 mg, 6.19 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour. The reaction solution was evaporated to dryness under reduced pressure to obtain a brown crude oil. The brown oil was subjected to silica gel column chromatography (dichloromethane:methanol = 7:1) to obtain compound 3k (105 mg, 88.5% yield) as a brown oil; LC-MS m / z (ESI): 288.2 [M+1] + .

[0367] Preparation of intermediate compound 4i

[0368] Step 1: Compound 3c (6 g, 34.06 mmol) and R-(+)-tert-butylsulfinamide (8.26 g, 68.12 mmol) were dissolved in anhydrous tetrahydrofuran (90 mL), and tetraethyl titanate (68.12 mmol, 14.28 mL) was added. Under argon protection, the reaction was heated to 75 °C and stirred for 20 hours until the dehydrated imine compound 4a was completely formed. The crude product was used directly in the reaction solution for the next step. LC-MS m / z (ESI): 280.1 [M+1] + .

[0369] Step 2: In a separate reaction flask, add tetrahydrofuran (40 mL) and sodium borohydride (5.15 g, 136.23 mmol). Lower the reaction temperature to -60°C. Slowly add the tetrahydrofuran mixture (90 mL) containing the imine product 4a from the previous step dropwise to the new reaction flask using a dropping funnel. After the addition is complete, gradually raise the reaction temperature to room temperature and react for 2 hours. After the reaction is complete, slowly add the reaction mixture to an excess of aqueous solution. A large amount of solid precipitates out, and bubbles are generated during the reaction. Take care to avoid bubbling. Filter through diatomaceous earth. Wash the filter cake with tetrahydrofuran. Extract the combined filtrates three times with ethyl acetate and twice with dichloromethane:methanol (10:1). Dry the combined organic phases with sodium sulfate and concentrate to obtain a brown solid. After column chromatography (dichloromethane:methanol = 20:1), compound 4b (4.8 g, 50.1% two-step reaction yield) is obtained as a brown solid. LC-MS m / z(ESI): 282.1 [M+1] + .

[0370] Step 3: Compound 4b (4.8 g, 17.06 mmol) was dissolved in anhydrous dioxane (20 mL), and dioxane hydrochloride (4 M, 21.32 mL) was added. The reaction mixture was stirred at room temperature for 0.5 hours. After concentration and evaporation to dryness, compound 4c (3.8 g, crude product) was obtained as a brown solid. This product was used directly in the next reaction. LC-MS m / z (ESI): 178.1 [M+1] + ;

[0371] Step 4: Compound 4c (3.5 g, 19.75 mmol) was dissolved in anhydrous tetrahydrofuran (40 mL) and water (40 mL). Sodium bicarbonate (6.64 g, 79.01 mmol) and BOC anhydride (8.62 g, 39.50 mmol) were slowly added in portions. The reaction mixture was stirred at room temperature for 20 hours. Ethyl acetate was added for extraction. The organic phases were combined, dried over sodium sulfate, and concentrated to obtain a crude oil. The crude oil was subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to give compound 4d (4.8 g, 87.6% yield) as a white solid. LC-MS m / z (ESI): 278.1 [M+1]+ .

[0372] Steps 5 to 9: Following the preparation method of intermediate compound 3k, replace starting material 3f with 4d to prepare intermediate 4i. LC-MS m / z (ESI): 288.1 [M+1] + .

[0373] Preparation of intermediate compound 12e

[0374] Step 1: A solution of 12a (2.9 g, 11.74 mmol), N-bromosuccinimide (2.19 g, 12.32 mmol), and azobisisobutyronitrile (96 mg, 0.59 mmol) in 1,2-dichloroethane (40 mL) was stirred at 85 °C for 5 hours. The reaction solution was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain a colorless liquid, which solidified upon standing to give a white solid 12b (2.6 g, 68.0% yield). 1 H NMR (400MHz, CDCl3) δ7.73~7.68(m,2H), 4.88(m,3H), 3.95(s,2H).

[0375] Step 2: 12b (2.6 g, 7.98 mmol), 3-amino-2,6-piperidinidone hydrochloride (1.71 g, 10.37 mmol), and diisopropylethylamine (3.09 g, 23.93 mmol) were dissolved in acetonitrile (40 mL). The reaction mixture was stirred at 80 °C for 48 hours. The reaction mixture was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to directly give gray solid 12c (2.7 g, 99.23% yield). MS m / z (ESI): 341.0 [M+1] + .

[0376] Step 3: A solution of 12c (2.7 g, 7.91 mmol), zinc cyanide (1.71 g, 7.91 mmol), 1,1'-bis(diphenylphosphine)ferrocene (877 mg, 1.58 mmol), and tris(dibenzylacetone)dipalladium (724 mg, 0.791 mmol) in N,N-dimethylformamide (30 mL) was stirred at 120 °C for 12 hours under nitrogen protection. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to give a gray solid 12d (2.0 g, 87.97% yield). LC-MS m / z (ESI): 288.0 [M+1] + .

[0377] Step 4: 12d (1 g, 3.48 mmol) was dissolved in anhydrous methanol (15 mL), and concentrated hydrochloric acid (1.5 mL) and platinum dioxide (237.3 mg, 1.04 mmol) were added. The reaction solution was stirred at room temperature for 12 h under a hydrogen atmosphere. The reaction solution was filtered, and the filter cake was washed with methanol (25 mL × 5). The filtered samples were combined and concentrated to give a yellow solid 12e (1.1 g, 96.41% yield). MS m / z (ESI): 292.1 [M+1] + .

[0378] Preparation of intermediate compound 13d

[0379] Step 1: Compound V1 (2.59 g, 10 mmol) was dissolved in N,N-dimethylformamide (20 mL), and ammonium chloride (2.1 g, 40 mmol), HATU (5.7 g, 1.5 mmol), and triethylamine (2 g, 20 mmol) were added. The mixture was reacted at room temperature for 1 hour, then extracted with water and ethyl acetate, and concentrated to dryness to give a brown solid 13a (2.7 g, crude product). LC-MS m / z (ESI): 259.0 [M+1] + .

[0380] Step 2: 13a (2.58 g, 10 mmol) was dissolved in tetrahydrofuran (15 mL), and boranetetrahydrofuran (60 mL) was added. The mixture was reacted at 70 °C for 2 hours, concentrated to dryness, quenched with methanol, and concentrated to dryness again to obtain a yellow oily substance 13b (2.9 g, crude product). LC-MS m / z (ESI): 245.0 [M+1] + .

[0381] Step 3: 13b (2.5 g, 10 mmol) was dissolved in tetrahydrofuran (40 mL), and di-tert-butyl dicarbonate (2.5 g, 12 mmol) and triethylamine (2.5 g, 25 mmol) were added. The mixture was reacted at room temperature for 12 hours, concentrated to dryness, and separated by silica gel column chromatography (ethyl acetate / petroleum ether = 31 vol% / 69 vol%) to give a yellow oil 13c (2.0 g, 56.8% yield). LC-MS m / z (ESI): 245.0 [M+1-100] + .

[0382] Step 4: 13c (0.7 g, 2 mmol) was dissolved in ethanol (10 mL), water (5 mL), and reduced iron powder (0.6 g, 10 mmol) and ammonium chloride (0.6 g, 10 mmol) were added. The mixture was reacted at 86 °C for 2 hours, filtered, and the filtrate was extracted and concentrated to dryness to obtain a yellow oily substance, 13d (0.5 g, crude product). LC-MS m / z (ESI): 215.0 [M+1-100] + .

[0383] Preparation of intermediate compound 16c

[0384] Step 1: 1-Benzyloxycarbonyl-3-pyrrolidone (412 mg, 1.88 mmol) and 2-(2-chloro-4-nitro-phenyl)ethanol (378.87 mg, 1.88 mmol) were dissolved in dichloromethane (8 mL). The mixture was stirred at 0 °C, and trimethylsilyl trifluoromethanesulfonate (1.29 g, 5.64 mmol) was added. The reaction was allowed to proceed for 0.5 hours, followed by the addition of triethylsilane (655.56 mg, 5.64 mmol). After the addition was complete, the mixture was stirred for another 0.5 hours. The mixture was then concentrated to dryness to obtain a yellow oily substance, 16b (0.8 g, crude product). LC-MS m / z (ESI): 405.0 [M+1] + .

[0385] Step 2: 16b (0.8 g, 2 mmol) was dissolved in ethanol (10 mL), water (5 mL), and reduced iron powder (0.6 g, 10 mmol) and ammonium chloride (0.6 g, 10 mmol) were added. The mixture was reacted at 86 °C for 2 hours, filtered, extracted with the filtrate, concentrated to dryness, and purified by flash column chromatography (12 g, 0-36 vol% ethyl acetate / petroleum ether) to obtain a yellow oily substance 16c (0.5 g, 67.5% yield). LC-MS m / z (ESI): 375.0 [M+1] + .

[0386] Preparation of intermediate compound 22f

[0387] Step 1: Compound 22a (491 mg, 2.02 mmol) and N-bromosuccinimide (431.37 mg, 2.42 mmol) were dissolved in dichloromethane (6 mL), stirred at room temperature, and then azobisisobutyronitrile (16.58 mg, 100.99 μmol) was added. The mixture was heated and stirred in a 90 °C oil bath under nitrogen protection for 2 hours. The reaction solution was evaporated to dryness under reduced pressure, and the crude product was purified by CombiFlash silica gel column chromatography to obtain compound 22b (570 mg, 87.64% yield). LC-MS m / z (ESI): 322.9 [M+1] + .

[0388] Step 2: Compound 22b (570 mg, 1.77 mmol) and 3-amino-2,6-piperidinedione (437.04 mg, 2.66 mmol, HCl) were dissolved in acetonitrile (5.0 mL), and then diisopropylethylamine (686.35 mg, 5.31 mmol, 925.00 μL) was added. Under nitrogen protection, the mixture was heated and stirred in an oil bath at 80 °C for 24 hours. After the reaction mixture cooled to room temperature, it was evaporated to dryness under reduced pressure. The crude product was purified by CombiFlash silica gel column chromatography to give compound 22c (171 mg, 28.65% yield). LC-MS m / z (ESI): 337.0 [M+1] + .

[0389] Step 3: Compound 22c (171 mg, 507.17 μmol) and zinc cyanide (59.55 mg, 507.17 μmol) were dissolved in DMF (2.5 mL), and 1,1'-bis(diphenylphosphine)ferrocene (14.06 mg, 25.36 μmol) was added. Under nitrogen protection, bis(dibenzylacetone)palladium (23.22 mg, 25.36 μmol) was added, and the mixture was heated and stirred in an oil bath at 100 °C for 14 hours. After the reaction solution cooled to room temperature, it was evaporated to dryness under reduced pressure. The crude product was purified by CombiFlash silica gel column chromatography to give compound 22d (133 mg, 92.57% yield). LC-MS m / z (ESI): 284.0 [M+1] + .

[0390] Step 4: Compound 22d (101 mg, 356.54 μmol) and di-tert-butyl dicarbonate (155.63 mg, 713.07 μmol) were dissolved in DMF (1.5 mL). The mixture was stirred in an ice-water bath at 0 °C under nitrogen protection. Nickel dichloride hexahydrate (338.98 mg, 1.43 mmol) and sodium borohydride (40.47 mg, 1.07 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred at room temperature for 16 hours. The reaction solution was evaporated to dryness under reduced pressure. The crude product was purified by CombiFlash silica gel column chromatography to give compound 22e (84 mg, 60.81% yield). LC-MS m / z (ESI): 388.2 [M+1] + .

[0391] Step 5: Compound 22e (84 mg, 216.81 μmol) was dissolved in dichloromethane (1.5 mL), stirred at 20 °C, and trifluoroacetic acid (247.21 mg, 2.17 mmol) was added. After the addition was complete, the reaction mixture was stirred for another 12 hours. The reaction solution was evaporated to dryness under reduced pressure. The remaining reaction solution was dissolved in ethyl acetate (30 mL) and acetonitrile (10 mL), and alkalized with triethylamine (1.5 mL). The solution was then evaporated to dryness under reduced pressure again. The crude product was purified by CombiFlash silica gel column chromatography to give compound 22f (30 mg, 48.16% yield). LC-MS m / z (ESI): 288.1 [M+1] + .

[0392] Preparation of intermediate compound 1.12A

[0393] A solution of 3-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)propionic acid (6.5 g, 29.38 mmol), (2R,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxane-4-yl)propionaldehyde (7.88 g, 29.38 mmol) in acetic acid (529 mg, 8.81 mmol) and methanol (100 mL) was added to a reaction flask. The reaction mixture was stirred at 50 °C for 0.5 h. Sodium cyanoborohydride (2.77 g, 44.07 mmol) was added in portions, and the reaction was stirred for another 2 h. Then, (2R,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy (2R,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxane-4-yl)propanal (7.88 g, 29.38 mmol) was added. The reaction mixture was stirred for 2 h. Sodium cyanoborohydride (2.77 g, 44.07 mmol) was added in portions, and the reaction mixture was stirred for 2 h. Sodium cyanoborohydride (2.77 g, 44.07 mmol) was added in portions, and the reaction mixture was stirred for 2 h. The reaction mixture was then stirred for 2 h. The reaction mixture was concentrated, and the solution was purified by preparative high-performance liquid chromatography (prep-HPLC). The white solid compound 1.12A (4.3 g, 20.17% yield) was obtained. MS m / z (ESI): 726.4 [M+1] + .

[0394] Preparation of intermediate compound 1.8A

[0395] Step 1: A solution of compound 1.8Aa (5 g, 29.38 mmol) in trifluoroacetic acid (5 mL) and dichloromethane (50 mL) was stirred at room temperature for 12 h. The reaction solution was then concentrated to give a yellow oily substance, 1.8Ab (5.15 g, 100% yield). MS m / z (ESI): 354.2 [M+1] + .

[0396] Step 2: Add 1.8Ab (5.05 g, 10.80 mmol) and a methanol (100 mL) solution of (2R,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxane-4-yl)propionaldehyde (2.9 g, 10.80 mmol) to the reaction flask. Stir the reaction mixture at 50 °C for 0.5 h. Add sodium cyanoborohydride (1.02 g, 16.21 mmol) in portions, and continue stirring for 2 h. Then add (2R,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxane-4-yl)propionaldehyde. (2R,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxane-4-yl)propanal (2.9 g, 10.80 mmol) was added, and the reaction mixture was stirred for 2 h. Sodium cyanoborohydride (1.02 g, 16.21 mmol) was added in portions, and the reaction mixture was stirred for 2 h. Sodium cyanoborohydride (1.02 g, 16.21 mmol) was added in portions, and the reaction mixture was stirred for 2 h. The reaction mixture was concentrated, and the solution was purified by preparative high-performance liquid chromatography (prep-HPLC). The white solid compound 1.8A (6.6 g, 71.21% yield) was obtained. MS m / z (ESI): 858.4 [M+1] + .

[0397] Preparation of intermediate compound 1.10A

[0398] Step 1: A methanol (50 mL) solution of compound 1.10Aa (4.9 g, 5.83 mmol) and Pd / C (750 mg, 10%) was stirred at room temperature for 12 h. The reaction solution was then concentrated to give a colorless oily substance 1.10Ab (4.7 g, 96.52% yield). MS m / z (ESI): 618.4 [M+1] + .

[0399] Step 2: Add 1.10Ab (4.56 g, 5.46 mmol), (2R,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxane-4-yl)propionaldehyde (1.47 g, 5.46 mmol), and acetic acid (984 mg, 16.39 mmol) in methanol (100 mL) to the reaction flask. Stir the reaction mixture at 50 °C for 0.5 h. Add sodium cyanoborohydride (686 mg, 16.21 mmol) in portions, and continue stirring for 2 h. Then add (2R,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl- 1,3-Dioxane-4-yl)propanal (1.47 g, 5.46 mmol) was added, and the reaction mixture was stirred for 2 h. Sodium cyanoborohydride (686 mg, 16.21 mmol) was added in portions, and the reaction mixture was stirred for another 2 h. Then, (2R,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxane-4-yl)propanal (1.47 g, 5.46 mmol) was added, and the reaction mixture was stirred for another 2 h. Sodium cyanoborohydride (686 mg, 16.21 mmol) was added in portions, and the reaction mixture was stirred for another 2 h. The reaction mixture was concentrated, and preparative high-performance liquid chromatography (prep-HPLC) was used for separation and purification. A yellow oily substance 1.10A (4.2 g, 68.51% yield) was obtained. MS m / z (ESI): 1122.6 [M+1] + .

[0400] Preparation of intermediate compound 1.24B

[0401] Compound 1.8A (5.4 g, 6.29 mmol) was dissolved in an aqueous HCl solution (60 mL, 2.4 M) and stirred for 2 hours at room temperature. The reaction mixture was concentrated to give compound 1.24B (4.3 g, 95.12% yield), which was used directly in the next reaction. LC-MS m / z (ESI): 682.1 [M+1] + .

[0402] Preparation of intermediate compound 1.26B

[0403] A 2M, 5mL solution of compound 1.12A (1.9g, 2.62mmol) in HCl was stirred at 50°C for 2 hours. The reaction mixture was then concentrated to give compound 1.26B (1.5g, 97.77% yield). LC-MS m / z (ESI): 550.3 [M+1] + .

[0404] Example 1: Preparation of Compound 1

[0405] Step 1: Dissolve V3 (328 mg, 1 mmol) in tetrahydrofuran (8 mL), add triphosgene (120 mg, 0.4 mmol), react at room temperature for 0.5 hours, evaporate to dryness, and then add to a solution of V2 (290 mg, 1 mmol), triethylamine (600 mg, 6 mmol), and N,N-dimethylformamide (5 mL). React at room temperature for 0.3 hours, concentrate to dryness, and separate by silica gel column chromatography (dichloromethane:methanol = 100:1 to 10:1) to directly obtain a yellow oil 1a (290 mg, 45.0% yield). LC-MS m / z (ESI): 546.0 [M+1-100] + .

[0406] Step 2: Dissolve 1a (65 mg, 0.1 mmol) in dichloromethane (5 mL), then add trifluoroacetic acid (2 mL) and react at room temperature (25 °C) for 0.5 hours. After the reaction, concentrate directly to obtain a yellow oil. The yellow oil was directly prepared and separated by HPLC (acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 10%-30%) and lyophilized to obtain a white solid compound 1 (18 mg, 40.8% yield). 1 H NMR (400MHz, DMSO-d6) δ10.91 (s, 1H), 9.00 (s, 1H), 7.63 (d, J = 2.0Hz, 1H), 7.59-7 .46(m,2H),7.17(dt,J=8.4,5.3Hz,2H),7.03(t,J=5.9Hz,1H),5.09(dd,J=13.3, 5.1Hz,1H),4.55(d,J=17.4Hz,1H),4.44-4.30(m,3H),3.60-3.51(m,4H),2.95-2 .78(m,5H),2.60(dd,J=18.8,16.5Hz,2H),2.46-2.31(m,4H),2.02-1.94(m,1H). LC-MS m / z (ESI): 546.2 [M+1] + .

[0407] Example 2: Preparation of Compound 2

[0408] Following the synthesis method of Example 1, starting material V3 was replaced with V4 to obtain the title product compound 2 (130 mg, 65% yield). 1H NMR (400MHz, DMSO-d6) δ10.99(s,1H),8.79(s,1H),7.64(d,J=2.1Hz,1H),7.57(d,J=7.7Hz,1H),7.53-7.47(m,1 H),7.19(d,J=8.4Hz,1H),7.12(dd,J=8.4,2.1Hz,1H),6.80(t,J=5.8Hz,1H),5.10(dd,J=13.3,5.3Hz,1H),4.55 (d,J=17.2Hz,1H),4.40(dd,J=19.9,11.5Hz,3H),3.52(t,J=7.0Hz,2H),3.42(t,J=5.6Hz,2H),3.28-3.11(m,2H ), 2.91 (d, J = 13.9Hz, 1H), 2.82 (t, J = 7.0Hz, 2H), 2.71 (s, 2H), 2.58 (d, J = 18.6Hz, 2H), 2.19 (s, 1H), 2.00 (s, 1H). LC-MS m / z(ESI):613.7[M+1] + .

[0409] Example 3: Preparation of Compound 3

[0410] Step 1: Dissolve intermediate V3 (90 mg, 273.69 μmol) in anhydrous dichloromethane (5 mL). Cool the reaction system to 0 °C. Under argon protection, add N,N-diisopropylethylamine (106.12 mg, 821.08 μmol, 143.01 μL). Then, slowly add triphosgene (26.80 mg, 90.32 μmol) dissolved in dichloromethane (1 mL) using a syringe. After maintaining the reaction in an ice-water bath for 1 hour, slowly add compound 3k (78.64 mg, 273.69 μmol) containing N,N-diisopropylethylamine (106.12 mg, 821.08 μmol, 143.01 μL) and dissolved in N,N-dimethylformamide (2 mL) using a syringe. Gradually increase the temperature to room temperature and react for 15 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product as a brown oil. After further evaporation, the brown oil solid was subjected to silica gel column chromatography (dichloromethane:methanol = 10:1) to give compound 3l (125 mg, 71.1% yield) as a brown oil. LC-MS m / z (ESI): 541.8 [M+1-100] + .

[0411] Step 2: Compound 3 (180 mg, 280.31 μmol) was dissolved in anhydrous dichloromethane (3 mL), and trifluoroacetic acid (1.15 g, 10.09 mmol, 750.00 μL) was added. The reaction temperature was gradually increased to room temperature and the reaction was carried out for 0.5 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product as a brown oil. The brown oil was evaporated to dryness and then sent to HPLC for preparation (acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 20%-50%). After separation, compound 3 (110 mg, 59.8% yield) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6) δ11.02(s,1H), δ9.12(s,1H),8.38(s,1H),7.69(d,J=7.8Hz,1H),7.63(d,J=2.1Hz, 1H),7.55(s,1H),7.48(d,J=7.9Hz,1H),7.39(s,1H),7.23-7.08(m,2H),5.11(dd,J=13.3,5.1Hz,1H),4.8 9(p,J=7.1Hz,1H),4.45(dd,J=17.4,2.9Hz,1H),4.30(dd,J=17.4,1.8Hz,1H),3.55(q,J=6.4,5.2Hz,4H), 3.01-2.73(m,5H),2.61(s,1H),2.41(s,3H),2.39-2.31(m,1H),2.04-1.90(m,1H),1.40(d,J=7.0Hz,3H). LC-MS m / z(ESI):542.2[M+1] + .

[0412] Example 4: Preparation of Compound 4

[0413] Following the preparation method of Example 3, starting material V3 was replaced with V4, and starting material 3k was replaced with 4i, to obtain the title product compound 4 (57.2 mg, 55.6% yield). 1H NMR (400MHz, DMSO-d6) δ11.00(s,1H),8.56(s,1H),7.70(d,J=7.9Hz,1H),7.70(d,J=2.1Hz,1H),7.55(s,1H),7.48(d ,J=7.9Hz,1H),7.19(d,J=8.4Hz,1H),7.09(dd,J=8.4,2.1Hz,1H),6.84(d,J=7.4Hz,1H),5.11(dd,J=13.3,5.2Hz,1H ),4.90(t,J=7.1Hz,1H),4.46(d,J=17.0Hz,1H),4.31(d,J=17.3Hz,1H),3.54(t,J=7.0Hz,2H),3.46(t,J=5.6Hz,2H) ,3.21(q,J=10.3Hz,2H),2.97-2.79(m,3H),2.76-2.55(m,4H),2.43-2.31(m,1H),2.00(s,1H),1.41(d,J=6.9Hz,3H). LC-MS m / z(ESI):610.1[M+1] + .

[0414] Example 5: Preparation of Compound 5

[0415] Following the preparation method of Example 4, the raw material R-(+)-tert-butylsulfinamide was replaced with S-(-)-tert-butylsulfinamide to obtain the title product compound 5 (48.3 mg, 32% yield). 1H NMR (400MHz, DMSO-d6) δ10.98(s,1H),8.56(s,1H),7.70(d,J=7.9Hz,1H),7.62(d,J=2.1Hz,1H),7.55(s,1H),7.48(d ,J=7.9Hz,1H),7.19(d,J=8.4Hz,1H),7.09(dd,J=8.4,2.1Hz,1H),6.83(d,J=7.4Hz,1H),5.10(dd,J=13.3,5.2Hz,1H ),4.90(t,J=7.1Hz,1H),4.45(d,J=17.0Hz,1H),4.31(d,J=17.3Hz,1H),3.53(t,J=7.0Hz,2H),3.43(t,J=5.6Hz,2H) ,3.21(q,J=10.3Hz,2H),2.97-2.79(m,3H),2.76-2.55(m,4H),2.43-2.31(m,1H),2.00(s,1H),1.41(d,J=6.9Hz,3H). LC-MS m / z(ESI):610.1[M+1] + .

[0416] Example 6: Preparation of Compound 6

[0417] Following the preparation method of Example 1, the starting material methylamine was replaced with 2,2-difluoroethylamine to obtain the title product compound 6 (71.93 mg, 17.39% yield). 1 H NMR (400MHz, DMSO-d6) δ11.01(s,1H),8.97(s,1H),7.65(d,J=2.0Hz,1H),7.57(d,J=7.7Hz,1H), 7.51(t,J=7.0Hz,1H),7.24-7.11(m,2H),7.00(t,J=6.0Hz,1H),6.09(tt,J=55.6,4.1Hz,1H),5. 11(dd,J=13.3,5.1Hz,1H),4.59-4.28(m,4H),3.61-3.47(m,4H),3.08(td,J=15.8,4.1Hz,2H),2 .95-2.77(m,6H),2.59(dd,J=15.4,2.7Hz,1H),2.41(td,J=13.2,4.5Hz,1H),2.05-1.94(m,1H). LC-MS m / z(ESI):596.2[M+1] + .

[0418] Example 7: Preparation of Compound 7

[0419] Following the preparation method of Example 1, the starting material methylamine was replaced with cyclopropylamine to obtain the title product compound 7 (4.8 mg, 15.0% yield). 1 H NMR (400MHz, DMSO-d6) δ11.00(s,1H),8.85(s,1H),7.64(d,J=2.0Hz,1H),7.57(d,J=7.7Hz,1H),7.54-7.48(m,1H),7.20(d,J=8 .4Hz,1H),7.14(dd,J=8.4,2.0Hz,1H),6.86(t,J=5.9Hz,1H),5.10(dd,J=13.3,5.1Hz,1H),4.42(d,J=5.7Hz,2H),4.37(d,J=17. 4Hz,1H),3.55(d,J=7.1Hz,2H),3.52(d,J=4.3Hz,1H),3.49(d,J=5.5Hz,2H),2.91(d,J=6.4Hz,1H),2.85(d,J=6.3Hz,4H),2.82( s,1H),2.58(d,J=17.4Hz,1H),2.42(dd,J=13.2,4.7Hz,1H),2.26(s,1H),2.02-1.96(m,1H),0.46(d,J=4.8Hz,2H),0.37(s,2H). LC-MS m / z(ESI):572.2[M+1] + .

[0420] Example 8: Preparation of Compound 8

[0421] Following the preparation method of Example 1, the starting material methylamine was replaced with (1R,2S)-2-fluorocyclopropane-1-amine to obtain the title product compound 8 (5.1 mg, 15.0% yield). 1H NMR (400MHz, DMSO-d6) δ11.00(s,1H),8.86(s,1H),7.76-7.68(m,1H),7.64(s,1H),7.57(d,J=7.6Hz, 1H),7.51(d,J=5.9Hz,1H),7.19(d,J=13.5Hz,1H),6.87(s,1H),5.17-5.02(m,1H),4.53(s,1H),4.40( d,J=8.4Hz,2H),3.76(d,J=34.9Hz,1H),3.62-3.57(m,2H),3.54-3.41(m,2H),3.07(s,2H),2.96(d,J =4.8Hz,2H),2.91-2.84(m,2H),2.83-2.70(m,2H),2.67-2.52(m,2H),2.47-2.29(m,2H),1.97(s,1H). LC-MS m / z(ESI):590.2[M+1] + .

[0422] Example 9: Preparation of Compound 9

[0423] Following the preparation method of Example 1, the starting material methylamine was replaced with 2-(methanesulfonyl)ethylamine to obtain the title product compound 9 (17.32 mg, 46.39% yield). 1 H NMR (400MHz, DMSO-d6) δ10.99(s,1H),8.83(s,1H),8.12(s,1H),7.63(d,J=2.0Hz,1H),7.57(d,J=7.7Hz,1H),7 .51(t,J=7.0Hz,1H),7.21(d,J=8.4Hz,1H),7.15(d,J=8.7Hz,1H),6.83(s,1H),5.09(dd,J=13.5,5.1Hz,1H),4. 55(d,J=17.3Hz,1H),4.42(d,J=5.8Hz,2H),4.37(d,J=17.3Hz,1H),3.57(d,J=7.0Hz,4H),3.24-3.17(m,3H),3. 06(s,3H),3.00(s,2H),2.87(d,J=7.2Hz,3H),2.59(d,J=17.2Hz,1H),2.04-1.94(m,2H),1.04(t,J=7.0Hz,1H). LC-MS m / z(ESI):638.2[M+1] + .

[0424] Example 10: Preparation of Compound 10

[0425] Following the preparation method of Example 1, the starting material methylamine was replaced with 2-aminoacetonitrile to obtain the title product compound 10 (50 mg, 37% yield). 1 H NMR (400MHz, DMSO-d6) δ10.99(s,1H),8.79(s,1H),7.64(d,J=2.1Hz,1H),7.57(d,J=7.7Hz,1H),7.53-7.47(m,1H),7 .19(d,J=8.4Hz,1H),7.12(dd,J=8.4,2.1Hz,1H),6.80(t,J=5.8Hz,1H),5.10(dd,J=13.3,5.3Hz,1H),4.55(d,J=17. 2Hz,1H),4.40(dd,J=19.9,11.5Hz,3H),3.52(t,J=7.0Hz,2H),3.42(t,J=5.6Hz,2H),3.28-3.11(m,1H),2.91(d,J=1 3.9Hz,1H),2.82(t,J=7.0Hz,2H),2.71(s,2H),2.66-2.54(m,2H),2.36(d,J=38.3Hz,1H),2.19(s,1H),1.97(s,1H). LC-MS m / z(ESI):571.2[M+1] + .

[0426] Example 11: Preparation of Compound 11

[0427] Following the preparation method of Example 1, the starting material methylamine was replaced with 3,3-difluorocyclobutylamine to obtain the title product compound 11 (130 mg, 65% yield). 1H NMR (400MHz, DMSO-d6) δ11.01(s,1H),8.85(s,1H),8.12(s,1H),7.63(s,1H),7.57(d,J=7.9Hz,1H),7 .51(d,J=6.8Hz,1H),7.22(d,J=8.6Hz,1H),7.16(d,J=10.5Hz,1H),6.85(s,1H),5.14-5.08(m,1H),4 .55(d,J=17.5Hz,1H),4.40(dd,J=20.2,11.7Hz,3H),3.65-3.57(m,3H),3.36(s,3H),3.06(s,2H),2. 90-2.84(m,2H),2.67(d,J=14.1Hz,2H),2.58(d,J=14.8Hz,1H),2.41(d,J=12.2Hz,3H),1.98(s,1H). LC-MS m / z(ESI):622.2[M+1] + .

[0428] Example 12: Preparation of Compound 12

[0429] Step 1: At 0°C, N,N'-dimethylformamide (2 mL) of N,N'-carbonyldiimidazole (29.59 mg, 182.46 μmol) was added in portions to a solution of compound V3 (60 mg, 182.46 μmol) and diisopropylethylamine (47.16 mg, 364.93 μmol, 63.56 μL). The mixture was slowly heated to room temperature and stirred for 1 hour. Then, the reaction solution was added dropwise to a solution of diisopropylethylamine (47.16 mg, 364.93 μmol, 63.56 μL) and 12e (53.15 mg, 182.46 μmol) of N,N-dimethylformamide (2 mL). The reaction solution was stirred for another 1 hour. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to give a yellow solid 12f (35 mg, 29.69% yield). MS m / z(ESI): 546.2 [M+1-100] + .

[0430] Step 2: At room temperature, a solution of 12f (35 mg, 54.17 μmol) in 0.2 mL of trifluoroacetic acid and 1 mL of dichloromethane was stirred for 1 hour. The reaction solution was concentrated and purified by preparative high performance liquid chromatography (acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 10%-65%) to give a white solid compound 12 (8.69 mg, 22.52% yield). 1H NMR (400MHz, DMSO-d6) δ10.99(s,1H),8.90(s,1H),8.39(brs,2H),7.65(d,J=4.0Hz,1H),7.5 8(d,J=4.0Hz,1H),7.53(d,J=12.0Hz,1H),7.23(d,J=4.0Hz,1H),7.17(dd,J=8.0,4.0Hz,1H), 6.89(t,J=4.0Hz,1H),5.10(dd,J=12.0,2.0Hz,1H),4.46-4.28(m,4H),3.63(t,J=8.0Hz,3H) ,3.10~3.08(m,2H),2.92~2.88(3H),2.60~2.55(m,4H),2.40~2.30(m,1H),2.02~1.97(m,1H). LC-MS m / z(ESI):546.2[M+1] + .

[0431] Example 13: Preparation of Compound 13

[0432] Step 1: Dissolve 13d (314 mg, 1 mmol) in tetrahydrofuran (8 mL), add triphosgene (120 mg, 0.4 mmol), react at room temperature for 0.5 hours, evaporate to dryness, and then add to a solution of compound V2 (290 mg, 1 mmol), triethylamine (600 mg, 6 mmol) in N,N-dimethylformamide (5 mL), react at room temperature for 0.3 hours, concentrate to dryness, and separate by silica gel column chromatography (dichloromethane:methanol = 100:1 to 10:1) to directly give a yellow oil 13e (350 mg, 55.5% yield). LC-MS m / z (ESI): 532.0 [M+1-100] + .

[0433] Step 2: Dissolve 13e (230 mg, 0.38 mmol) in dichloromethane (5 mL), then add trifluoroacetic acid (2 mL) and react at room temperature (25 °C) for 0.5 hours. After the reaction is complete, directly evaporate to dryness and concentrate to obtain a yellow oily substance 13f (390 mg, crude product). LC-MS m / z (ESI): 532.0 [M+1] + .

[0434] Step 3: Dissolve 13f (390 mg, 0.7 mmol) in dichloromethane (3 mL) and acetic acid (3 mL), then add 3-oxetane (400 mg, 5.6 mmol), react at 60 °C for 2 hours, add sodium borohydride acetate (400 mg, 2.3 mmol), react at 60 °C for 2 hours, concentrate to dryness, and separate by silica gel column chromatography (dichloromethane:methanol = 100:1 to 8:1) to obtain a yellow oily substance. Then send it to preparative high performance liquid chromatography (acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 5%-35%). After freeze-drying, obtain a white solid compound 13 (60 mg, 15% yield). 1 H NMR (400MHz, DMSO-d6) δ11.01(s,1H),8.85(s,1H),8.43(s,1H),7.63(s,1H),7.57(d,J=7.9Hz,1H),7 .51(d,J=6.8Hz,1H),7.22(d,J=8.6Hz,1H),7.16(d,J=10.5Hz,1H),6.85(s,1H),5.14-5.08(m,1H),4 .55(d,J=17.5Hz,1H),4.40(dd,J=20.2,11.7Hz,3H),3.65-3.57(m,3H),3.36(s,2H),3.06(s,3H),2. 90-2.84(m,2H),2.67(d,J=14.1Hz,2H),2.58(d,J=14.8Hz,1H),2.41(d,J=12.2Hz,3H),1.98(s,1H). LC-MS m / z(ESI):588.2[M+1] + .

[0435] Example 14: Preparation of Compound 14

[0436] Following the preparation method of Example 13, the starting material 3-oxetane was replaced with cyclobutanone to obtain the title product compound 14 (15 mg, 14% yield). 1H NMR (400MHz, DMSO-d6) δ11.00(s,1H),8.89(s,1H),7.64(s,1H),7.56(d,J=7.7Hz ,1H),7.50(s,1H),7.19(s,1H),7.15(s,1H),6.90(s,1H),5.10(d,J=8.4Hz,1H), 4.55(d,J=17.0Hz,1H),4.39(dd,J=19.4,11.2Hz,3H),3.51(s,4H),3.13(s,4H), 2.82(s,3H),2.63(d,J=20.0Hz,2H),2.43-2.39(m,1H),2.00(s,3H),1.58(s,3H). LC-MS m / z (ESI): 586.2 [M+1] + .

[0437] Example 15: Preparation of Compound 15

[0438] Following the preparation method of Example 13, the starting material 3-oxetane was replaced with (3,3-difluorocyclobutyl)formaldehyde to obtain the title product compound 15 (15 mg, 14% yield). 1 H NMR (400MHz, DMSO-d6) δ11.01(s,1H),8.85(s,1H),8.43(s,1H),7.63(s,1H),7.57(d,J=7.9Hz,1H),7 .51(d,J=6.8Hz,1H),7.22(d,J=8.6Hz,1H),7.16(d,J=10.5Hz,1H),6.85(s,1H),5.14-5.08(m,1H),4 .55(d,J=17.5Hz,1H),4.40(dd,J=20.2,11.7Hz,3H),3.65-3.57(m,3H),3.36(s,4H),3.06(s,3H),2. 90-2.84(m,2H),2.67(d,J=14.1Hz,2H),2.58(d,J=14.8Hz,1H),2.41(d,J=12.2Hz,3H),1.98(s,1H). LC-MS m / z(ESI):636.2[M+1] + .

[0439] Example 16: Preparation of Compound 16

[0440] Step 1: 16c (188 mg, 0.5 mmol) was dissolved in tetrahydrofuran (6 mL), followed by the addition of triphosgene (60 mg, 0.2 mmol). The mixture was reacted at room temperature for 0.5 h, evaporated to dryness, and then added to a solution of V2 (145 mg, 0.5 mmol) and triethylamine (300 mg, 3 mmol) in N,N-dimethylformamide (5 mL). The mixture was reacted at room temperature for 0.3 h, concentrated to dryness, and separated by silica gel column chromatography (dichloromethane:methanol = 100:1 to 10:1) to directly give a yellow oily substance 16d (180 mg, 51.8% yield). LC-MS m / z (ESI): 692.0 [M+1] + .

[0441] Step 2: Dissolve 16d (180 mg, 0.26 mmol) in methanol (5 mL), then add palladium-carbon (40 mg), and react at room temperature (25 °C) for 0.5 h under hydrogen atmosphere. After the reaction is complete, filter directly, and concentrate by rotary evaporation to obtain a yellow oil. The yellow oil was directly sent to preparative high performance liquid chromatography (acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 5%-30%) for separation. After freeze-drying, a white solid compound 16 (30 mg, 20.8% yield) was obtained. 1 H NMR (400MHz, DMSO-d6) δ9.42(s,1H),8.40(s,1H),7.65(s,1H),7.56(d,J=7.8Hz,1H),7.53-7.45(m,2H),7.19(s,2H),5.09(d,J=13.1Hz,1H), 4.55(d,J=17.4Hz,1H),4.45-4.29(m,4H),3.77(s,2H),3.53(dd,J=21.4,6.2Hz,7H),2.91-2.82(m,4H),2.58(d,J=17.2Hz,1H),1.99(s,1H). LC-MS m / z(ESI):558.2[M+1] + .

[0442] Example 17: Preparation of Compound 17

[0443] Following the preparation method of Example 16, the starting material 1-benzyloxycarbonyl-3-pyrrolidone was replaced with N-benzyloxycarbonyl-2-pyrrolidinecarboxaldehyde to obtain the title product compound 17 (43 mg, 33.8% yield). 1H NMR(400MHz,DMSO-d6)δ11.11-10.89(m,1H),9.48(s,1H),8.37(s,1H),7.65(s,1H),7.61- 7.47(m,3H),7.19(s,2H),5.10(dd,J=13.2,5.0Hz,1H),4.55(d,J=17.2Hz,1H),4.44-4.31 (m,3H),3.57(s,2H),3.49-3.46(m,2H),3.41(s,2H),2.95(d,J=6.9Hz,3H),2.84(t,J=6.9 Hz, 2H), 2.58 (d, J = 15.0Hz, 1H), 2.00 (s, 1H), 1.85 (s, 1H), 1.78-1.62 (m, 2H), 1.43 (s, 1H). LC-MS m / z(ESI):572.2[M+1] + .

[0444] Example 18: Preparation of Compound 18

[0445] Following the preparation method of Example 16, the starting material 1-benzyloxycarbonyl-3-pyrrolidone was replaced with benzyl 3-formylaziridine-1-carboxylic acid benzyl ester to obtain the title product compound 18 (25 mg, 16.8% yield). 1 H NMR (400MHz, DMSO-d6) δ9.42(s,1H),8.40(s,1H),7.65(s,1H),7.56(d,J=7.8Hz,1H),7.53-7.45(m,2H),7.19(s,2H),5.09(d,J=13.1Hz,1H), 4.55(d,J=17.4Hz,1H),4.45-4.29(m,4H),3.77(s,2H),3.53(dd,J=21.4,6.2Hz,7H),2.91-2.82(m,4H),2.58(d,J=17.2Hz,1H),1.99(s,1H). LC-MS m / z(ESI):558.2[M+1] + .

[0446] Example 19: Preparation of Compound 19

[0447] Following the preparation method of Example 16, the starting material 1-benzyloxycarbonyl-3-pyrrolidone was replaced with N-Cbz-piperidin-3-one to obtain the title product compound 19 (4.15 mg, 8.68% yield). 1H NMR (400MHz, DMSO-d6) δ11.01(s,1H),8.92(s,1H),8.26(s,1H),7.63(d,J=2.1Hz,1H),7.57(d,J=7.7Hz,1H),7.53-7.48(m ,1H),7.21(d,J=8.4Hz,1H),7.15(dd,J=8.6,2.2Hz,1H),6.93(t,J=6.0Hz,1H),5.10(dd,J=13.3,5.1Hz,1H),4.55(d,J=17 .4Hz,1H),4.42(d,J=5.8Hz,2H),4.37(d,J=17.4Hz,1H),3.57(t,J=7.2Hz,2H),3.48-3.40(m,2H),3.04(d,J=13.6Hz,1H), 2.82(t,J=7.0Hz,2H),2.75-2.66(m,2H),2.63-2.54(m,2H),1.98(d,J=13.6Hz,1H),1.77(s,1H),1.68(s,1H),1.44(s,3H). LC-MS m / z(ESI):571.8[M+1] + .

[0448] Example 20: Preparation of Compound 20

[0449] Following the preparation method of Example 16, the starting material 1-benzyloxycarbonyl-3-pyrrolidone was replaced with 1-benzyloxycarbonyl-3-azacyclobutanone to obtain the title product compound 20 (4.54 mg, 3.84% yield). 1 H NMR (400MHz, DMSO-d6) δ10.97(s,1H),9.16(s,1H),8.33(s,1H),7.64(s,1H),7.56(d,J=7.7Hz, 1H),7.54-7.48(m,1H),7.22-7.12(m,3H),5.10(dd,J=13.3,5.0Hz,1H),4.55(d,J=17.3Hz,1H) ,4.41(d,J=5.9Hz,2H),4.37(d,J=17.4Hz,1H),3.72(s,2H),3.53-3.44(m,4H),2.95-2.85(m,1 H), 2.83-2.78 (m, 2H), 2.58 (d, J = 16.8Hz, 2H), 2.42 (dd, J = 13.1, 4.5Hz, 1H), 2.03-1.93 (m, 1H). LC-MS m / z(ESI):544.1[M+1] + .

[0450] Example 21: Preparation of compound 21

[0451] Step 1: 13f (531 mg, 1 mmol) was dissolved in dichloromethane (10 mL) and methanol (3 mL), followed by the addition of 2-oxoacetic acid (740 mg, 10 mmol) and Ti(OiPr)4 (0.8 mL). The reaction was carried out at 60 °C for 2 h, followed by the addition of sodium borohydride acetate (750 mg, 3 mmol), and then the addition of 2-oxoacetic acid (740 mg, 10 mmol). The reaction was carried out at 60 °C for 4 h, followed by the addition of water (5 mL), filtration, washing with dichloromethane / methanol, concentration, and separation by silica gel column chromatography (dichloromethane:methanol = 100:1 to 3:1) to directly obtain an oily product (210 mg). This product was then sent for preparative high performance liquid chromatography (acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 5%-25%). After freeze-drying, the title compound 21 (63 mg, 11% yield) was obtained. 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),10.98(s,1H),7.64(d,J=2.1Hz,1H),7.57(d,J=7.7Hz,1H),7.53-7.47(m,1H), 7.19(d,J=8.4Hz,1H),7.12(dd,J=8.4,2.1Hz,1H),6.80(t,J=5.8Hz,1H),5.10(dd,J=13.3,5.3Hz,1H),4.55(d,J=17 .2Hz,1H),4.40(dd,J=19.9,11.5Hz,3H),3.52(t,J=7.0Hz,2H),3.42(t,J=5.6Hz,2H),3.28-3.11(m,2H),2.91(d,J= 13.9Hz,1H),2.82(t,J=7.0Hz,2H),2.71(s,2H),2.66-2.54(m,2H),2.36(d,J=38.3Hz,1H),2.19(s,1H),1.97(s,1H). LC-MS m / z(ESI):544.1[M+1] + .

[0452] Example 22: Preparation of compound 22

[0453] Following the synthesis method of Example 1, starting material V2 was replaced with compound 22f to obtain the title product compound 22 (19.26 mg, 58.95% yield). 1H NMR (400MHz, DMSO-d6) δ10.99(s,1H),8.92(s,1H),8.30(s,1H),7.65(d,J=2.1Hz,1H),7.53(d,J=7.8Hz,1 H),7.40(d,J=7.9Hz,1H),7.24-7.12(m,2H),6.93(t,J=5.6Hz,1H),5.11(dd,J=13.3,5.1Hz,1H),4.42(d, J=17.1Hz,1H),4.37(d,J=5.7Hz,2H),4.25(d,J=17.1Hz,1H),3.56(q,J=5.7,4.4Hz,4H),2.88(dt,J=19.6 ,6.1Hz,4H),2.63-2.56(m,1H),2.43(s,3H),2.38(dd,J=13.3,8.8Hz,2H),2.27(s,3H),2.03-1.94(m,1H). LC-MS m / z(ESI):541.8[M+1] + .

[0454] Example 23: Preparation of compounds 23-1 and 23-2

[0455] Following the preparation method of Example 1, the starting material methylamine was replaced with (S)-1,1,1-trifluoropropane-2-amine to obtain the title product compound 23-1 (26.52 mg, 22.74% yield). 1 H NMR (400MHz, DMSO-d6) δ11.01(s,1H),8.93(s,1H),8.14(s,1H),7.65(d,J=8.0,1H),7.58(d,J=8.0,1H,1H),7.5 2~7.51(m,1H),7.21~7.19(d,J=8.0,1H),7.16~7.13(m,1H),6.96~6.93(m,1H),5.11(dd,J=16.0,8.0Hz,1H),4. 56(d,J=20.0Hz,1H),4.44~4.37(m,3H),3.56~3.52(m,2H),3.44~3.40(m,2H),3.26(brs,1H),2.96~2.82(m,3H) ,2.73(brs,2H),2.58~2.51(m,1H),2.49~2.42(m,1H),2.06~1.98(m,1H),1.82~1.78(m,1H),1.10~1.08(m,2H). LC-MS m / z(ESI):628.2[M+1] + .

[0456] Following the preparation method of Example 1, the starting material methylamine was replaced with (R)-1,1,1-trifluoropropane-2-amine to obtain the title product compound 23-2 (22.52 mg, 22.72% yield). 1 H NMR (400MHz, DMSO-d6) δ11.01(s,1H),8.85(s,1H),8.14(s,1H),7.66(s,1H),7.65~7.57(m,1H),7.54~7.51(m, 1H),7.21~7.19(d,J=8.0,1H),7.15~7.13(m,1H),6.87~6.85(m,1H),5.13(dd,J=12.0,4.0Hz,1H),4.56(d,J=2 0.0Hz,1H),4.44~4.37(m,3H),3.56~3.52(m,2H),3.44~3.40(m,2H),3.28~3.23(m,1H),2.91~2.82(m,3H),2.7 6~2.70(m,2H),2.58~2.51(m,1H),2.46~2.42(m,1H),2.02~1.98(m,1H),1.99~1.83(m,1H),1.10~1.06(m,2H). LC-MS m / z(ESI):628.2[M+1] + .

[0457] Example 24: Preparation of compound 24

[0458] Following the synthesis method of Example 1, starting material V2 was replaced with 12e and starting material V3 was replaced with V4 to obtain the title product compound 24 (130 mg, 65% yield). 1H NMR (400MHz, DMSO-d6) δ10.99(s,1H),8.87(s,1H),8.32(s,1H),7.65(d,J=2.1Hz,1H),7.56(d,J=6.3Hz,1H),7.50(d,J=8. 8Hz,1H),7.19(d,J=8.4Hz,1H),7.12(dd,J=8.2,2.2Hz,1H),6.85(t,J=6.0Hz,1H),5.09(dd,J=13.3,5.0Hz,1H),4.46-4.4 1(m,1H),4.40(s,2H),4.28(d,J=17.3Hz,1H),3.52(t,J=7.0Hz,2H),3.42(t,J=5.8Hz,2H),3.20(q,J=10.6Hz,2H),2.82(t ,J=7.1Hz,2H),2.70(t,J=5.6Hz,2H),2.67-2.63(m,1H),2.62-2.58(m,1H),2.35(d,J=13.2Hz,1H),1.98(d,J=9.7Hz,1H). LC-MS m / z(ESI):614.2[M+1] + .

[0459] Example 25: Preparation of Compound 25

[0460] Step 1: Compound 21 (20 mg, 0.03 mmol) was dissolved in DMF (2 mL), and ammonium chloride (28 mg, 0.5 mmol), HATU (18 mg, 0.045 mmol), and triethylamine (11 mg, 0.1 mmol) were added. The mixture was reacted at room temperature for 1 hour, concentrated to dryness, and directly sent to preparative high performance liquid chromatography for separation (acid method, GILSON, Waters-SunFire C18; ACN / Water + 0.04% FA; gradient, 10%-30%). After freeze-drying, compound 25 (6 mg, 30.1% yield) was obtained. 1H NMR (400MHz, DMSO-d6) δ10.99(s,1H),7.64(d,J=2.1Hz,1H),7.57(d,J=7.7Hz,1H),7.53-7.47(m,1H),7.19(d,J=8.4 Hz,1H),7.25(s,2H),,7.12(dd,J=8.4,2.1Hz,1H),6.80(t,J=5.8Hz,1H),5.10(dd,J=13.3,5.3Hz,1H),4.55(d,J=17 .2Hz,1H),4.40(dd,J=19.9,11.5Hz,3H),3.52(t,J=7.0Hz,2H),3.42(t,J=5.6Hz,2H),3.28-3.11(m,2H),2.91(d,J= 13.9Hz,1H),2.82(t,J=7.0Hz,2H),2.71(s,2H),2.66-2.54(m,2H),2.36(d,J=38.3Hz,1H),2.19(s,1H),1.98(s,1H). LC-MS m / z(ESI):589.2[M+1] + .

[0461] Example 26: Preparation of Compound 26

[0462] Following the synthesis method of Example 1, the starting material methylamine was replaced with 2-aminoethanesulfonamide to obtain the title product compound 26 (23.43 mg, 29.46% yield). 1 H NMR (400MHz, DMSO-d6) δ11.01(s,1H),8.84(s,1H),8.14(s,1H),7.66(s,1H),7.65~7.51(m, 2H),7.22(d,J=8.0Hz,1H),7.15(d,J=8.0Hz,1H),6.91~6.83(m,3H),5.11(dd,J=16.0,8.0H z,1H),4.56(d,J=20.0Hz,1H),4.48~4.31(m,3H),3.58~3.49(m,4H),3.20~3.16(m,2H),3.0 3~2.92(m,2H),2.88~2.81(m,5H),2.58~2.51(m,1H),2.42~2.41(m,1H),2.02~1.99(m,1H). LC-MS m / z(ESI):639.2[M+1] + .

[0463] Example 27: Preparation of Compound 27

[0464] Following the preparation method of Example 4, the starting material R-(+)-tert-butylsulfinamide was replaced with S-(-)-tert-butylsulfinamide, and the starting material V4 was replaced with V3, to obtain the title product compound 27 (45.3 mg, 32% yield). 1 H NMR (400MHz, DMSO-d6) δ9.04(s,1H),8.34(s,1H),7.69(d,J=7.9Hz,1H),7.63(d,J=2.0Hz,1 H),7.55(s,1H),7.48(d,J=7.9Hz,1H),7.30(s,1H),7.22-7.10(m,2H),5.10(dd,J=13.1,5.0 Hz,1H),4.89(t,J=7.1Hz,1H),4.45(dd,J=17.4,3.0Hz,1H),4.31(d,J=17.3Hz,1H),3.55(q, J=6.6,5.9Hz,6H),3.01-2.54(m,6H),2.40(s,3H),1.99(s,1H),1.40(d,J=7.0Hz,3H).LC-MS m / z(ESI):542.2[M+1] + .

[0465] Example 28: Preparation of compound 28

[0466] Following the preparation method of Example 1, the starting material methylamine was replaced with ethylamine to obtain the title product compound 28 (187 mg, 33.32% yield). 1 H NMR(400MHz,DMSO-d6)δ10.97(s,1H),9.53(s,1H),8.44(s,1H),7.66(s,1H),7.61(s,1H) ,7.57-7.48(m,2H),7.19(s,2H),5.09(dd,J=13.2,5.0Hz,1H),4.54(d,J=17.4Hz,1H),4. 39(t,J=12.9Hz,3H),3.58(dd,J=12.2,5.1Hz,4H),2.98(s,2H),2.91-2.79(m,5H),2.59( d, J=16.0Hz, 1H), 2.42 (dd, J=13.1, 4.2Hz, 1H), 2.06-1.94 (m, 1H), 1.11 (t, J=7.1Hz, 3H). LC-MS m / z(ESI):560.0[M+1] + .

[0467] Example 29: Preparation of compound 29

[0468] Following the preparation method of Example 1, the starting material methylamine was replaced with ethanolamine to obtain the title product compound 29 (340 mg, 29.63% yield). 1 H NMR(400MHz,DMSO-d6)δ11.03(s,1H),9.23(d,J=11.4Hz,1H),8.29(s,1H),7.71-7.46 (m,3H),7.35-7.13(m,3H),5.11(dd,J=13.3,5.1Hz,1H),4.57(d,J=17.4Hz,1H),4.48- 4.33(m,3H),3.61-3.46(m,7H),2.99-2.80(m,5H),2.73(d,J=4.9Hz,2H),2.60(ddd,J= 17.4, 4.4, 2.2Hz, 1H), 2.43 (dd, J=13.2, 4.5Hz, 1H), 2.00 (dd, J=9.3, 4.3Hz, 1H); LC-MS m / z(ESI):576.2[M+1] +

[0469] Example 30: Preparation of compound 30

[0470] Following the preparation method of Example 1, the starting material methylamine was replaced with (R)-N-benzyloxycarbonyl-2-pyrrolidinecarboxaldehyde to obtain the title product compound 30 (26.70 mg, 30.27% yield). 1 H NMR(400MHz,DMSO-d6)δ9.74(d,J=4.9Hz,1H),8.45(s,1H),7.99(dd,J=7.7,4.3Hz,1H),7.69-7.60 (m,2H),7.54(s,1H),7.46(d,J=7.9Hz,1H),7.17(d,J=3.7Hz,2H),5.08(dd,J=13.3,5.1Hz,1H),4.8 7(t,J=7.1Hz,1H),4.42(dd,J=17.3,4.9Hz,1H),4.27(dd,J=17.3,2.1Hz,1H),3.61-3.39(m,6H),3 .07-2.77(m,5H),2.63-2.52(m,1H),2.43-2.27(m,1H),2.01-1.67(m,4H),1.52-1.29(m,4H); LC-MS m / z(ESI):568.3[M+1] +

[0471] Example 31: Preparation of compound 31

[0472] Following the preparation method of Example 27, the starting material methylamine was replaced with 2-(methanesulfonyl)ethylamine to obtain the title product compound 31 (140 mg, 73.33% yield). 1 H NMR (400MHz, DMSO-d6) δ11.00(s,1H),8.61(d,J=3.5Hz,1H),8.15(s,1H),7.70(d,J=7.9Hz,1H),7.62(d,J=2.1Hz,1H),7.55(s,1H),7.48( d,J=7.9Hz,1H),7.20(d,J=8.4Hz,1H),7.10(dd,J=8.4,2.2Hz,1H),6.88(dd,J=7.8,2.9Hz,1H),5.11(dd,J=13.3,5.1Hz,1H),4.89(p,J=6 .9Hz,1H),4.45(dd,J=17.5,2.0Hz,1H),4.31(d,J=17.3Hz,1H),3.63-3.44(m,4H),3.25(t,J=6.9Hz,2H),3.00(d,J=7.6Hz,5H),2.96-2.8 0(m,3H),2.75(t,J=5.4Hz,2H),2.59(d,J=17.4Hz,1H),2.45-2.28(m,1H),1.98(ddd,J=9.9,5.2,2.6Hz,1H),1.41(d,J=6.9Hz,3H); LC-MS m / z(ESI):634.2[M+1] +

[0473] Example 32: Preparation of compound 32

[0474] Following the preparation method of Example 27, the starting material methylamine was replaced with isopropylamine to obtain the title product compound 32 (54 mg, 30.32% yield). 1H NMR (400MHz, DMSO-d6) δ10.99(s,1H),9.22(s,1H),8.36(s,1H),7.67(d,J=7.7Hz,1H),7.62(s,1H),7.53(s, 1H),7.46(d,J=7.9Hz,2H),7.16(t,J=8.3Hz,2H),5.09(dd,J=13.4,4.7Hz,1H),4.92-4.84(m,1H),4.43(d,J= 17.7Hz,1H),4.28(d,J=16.8Hz,1H),3.52(dd,J=15.0,7.8Hz,4H),2.89(s,1H),2.81(dd,J=15.0,7.8Hz,4H) ,2.58(dd,J=36.5,19.1Hz,2H),2.41-2.28(m,1H),1.98(s,1H),1.38(d,J=6.9Hz,3H),1.01(d,J=6.0Hz,6H). LC-MS m / z(ESI):570.2[M+1] +

[0475] Example 33: Preparation of compound 33

[0476] Following the preparation method of Example 27, the starting material methylamine was replaced with ethylamine to obtain the title product compound 33 (231 mg, 43.20% yield). 1 H NMR (400MHz, DMSO-d6) δ10.99(brs,1H),9.36(s,1H),8.38(s,1H),7.66(d,J=7.9Hz,1H),7.62(s,2H),7.54(s,1 H),7.46(d,J=7.8Hz,1H),7.16(s,2H),5.09(dd,J=13.3,5.1Hz,1H),4.87(t,J=7.0Hz,1H),4.42(dd,J=17.2,4.0 Hz,1H),4.28(d,J=17.3Hz,1H),3.55(t,J=5.6Hz,5H),2.89(d,J=4.9Hz,3H),2.83(t,J=6.7Hz,2H),2.78-2.73(m ,2H),2.57(d,J=17.4Hz,1H),2.40-2.31(m,1H),2.01-1.92(m,1H),1.38(d,J=6.9Hz,3H),1.07(t,J=6.5Hz,3H). LC-MS m / z(ESI):556.2[M+1] +

[0477] Example 34: Preparation of compound 34

[0478] Following the preparation method of Example 27, the starting material methylamine was replaced with (S)-3-aminotetrahydrofuran to obtain the title product compound 34 (32 mg, 28.23% yield). 1 H NMR (400MHz, DMSO-d6) δ10.97(s,1H),8.85(s,1H),8.21(s,1H),7.67(d,J=7.8Hz,1H),7.61(d,J=1.8Hz,1H),7 .53(s,1H),7.46(d,J=7.9Hz,1H),7.17(d,J=8.4Hz,1H),7.14-7.08(m,2H),5.09(dd,J=13.3,4.9Hz,1H),4.91-

[0479] 4.84(m,1H),4.43(dd,J=17.4,2.3Hz,1H),4.29(d,J=17.3Hz,1H),3.73(dd,J=14.8,7.6Hz,1H),3.67-3.6 3(m,1H),3.60(dd,J=7.9,6.1Hz,1H),3.54(dd,J=12.6,5.6Hz,2H),3.48-3.43(m,3H),3.39(d,J=10.7Hz, 1H),2.90(dd,J=10.6,6.6Hz,1H),2.82(t,J=6.9Hz,2H),2.73(dt,J=12.5,6.1Hz,2H),2.58(d,J=17.1Hz, 1H),2.37(d,J=13.0Hz,1H),2.01-1.92(m,2H),1.66(dd,J=11.1,5.7Hz,1H),1.39(d,J=6.9Hz,3H).LC-MS m / z(ESI):598.0[M+1] +

[0480] Example 35: Preparation of compound 35

[0481] Following the preparation method of Example 27, the starting material methylamine was replaced with (R)-3-aminotetrahydrofuran to obtain the title product compound 35 (102.83 mg, 39.52% yield). 1H NMR (400MHz, DMSO-d6) δ10.99(s,1H),8.73(d,J=3.8Hz,1H),8.19(s,1H),7.68(d,J=7.8 Hz,1H),7.61(d,J=2.1Hz,1H),7.54(s,1H),7.47(d,J=7.9Hz,1H),7.18(d,J=8.4Hz,1H) ,7.09(dd,J=8.4,2.2Hz,1H),7.00(dd,J=7.7,3.3Hz,1H),5.09(dd,J=13.3,5.1Hz,1H), 4.88(p,J=6.9Hz,1H), 4.43(dd,J=17.3,2.8Hz,1H), 4.29(d,J=17.3Hz,1H), 3.71(q,J=7. 5Hz,1H),3.65(dd,J=9.0,5.9Hz,1H),3.59(td,J=8.0,5.7Hz,1H),3.52(t,J=7.0Hz,2H) ,3.45(t,J=5.5Hz,2H),3.43-3.40(m,1H),3.33(td,J=6.8,3.8Hz,1H),2.90(ddd,J=17. 2,13.6,5.4Hz,1H),2.82(t,J=7.0Hz,2H),2.71(dq,J=11.8,5.7Hz,2H),2.62-2.54(m,1 H),2.44-2.29(m,1H),2.02-1.87(m,2H),1.68-1.58(m,1H),1.39(d,J=6.9Hz,3H); LC-MS m / z(ESI):598.3[M+1] + .

[0482] Example 36: Preparation of compound 36

[0483] Following the preparation method of Example 27, the starting material methylamine was replaced with 2-aminoethylsulfonamide to obtain the title product compound 36 (285.97 mg, 41.69% yield). 1H NMR (400MHz, DMSO-d6) δ10.98(s,1H),8.58(s,1H),8.16(s,1H),7.70(d,J=7.9Hz,1H),7.62(d,J=2.2Hz,1H),7.55(s,1H),7.48(d,J =7.9Hz,1H),7.20(d,J=8.4Hz,1H),7.10(d,J=8.8Hz,1H),6.86(s,2H),5.10(dd,J=13.3,5.0Hz,1H),4.90(t,J=7.1Hz,1H),4.45(d, J=17.3Hz,1H),4.31(d,J=17.3Hz,1H),3.53(t,J=7.1Hz,2H),3.45(t,J=5.5Hz,2H),3.34-3.30(m,1H),3.10(d,J=7.0Hz,2H),2.92( s,3H),2.83(t,J=7.1Hz,2H),2.68(s,2H),2.60(d,J=16.9Hz,1H),2.39(d,J=14.0Hz,1H),2.00(s,1H),1.42(d,J=6.9Hz,3H); LC-MS m / z(ESI):635.2[M+1] + .

[0484] Example 37: Preparation of compound 37

[0485] Following the preparation method of Example 1, the starting material methylamine was replaced with 1,1-dioxide-3-aminothiohexacyclobutane to obtain the title product compound 37 (315 mg, 24.65% yield). 1H NMR (400MHz, DMSO-d6) δ11.00(s,1H),8.80(s,1H),7.68-7.62(m,1H),7.61-7.44(m,2H),7.22(d,J=8.4Hz,1H) ,7.15(dd,J=8.3,2.2Hz,1H),6.82(t,J=5.9Hz,1H),5.11(dd,J=13.3,5.0Hz,1H),4.57(d,J=17.3Hz,1H),4.47- 4.33(m,3H),4.25(dd,J=14.1,8.2Hz,2H),3.86(dd,J=14.0,5.0Hz,2H),3.54(t,J=7.0Hz,3H),3.44(t,J=5.5H LC-MS m / z(ESI):636.2[M+1] + .

[0486] Example 38: Preparation of compound 38

[0487] Following the preparation method of Example 1, the starting material methylamine was replaced with 3-aminotetrahydrothiophene 1,1-dioxide to obtain the title product compound 38 (127.64 mg, 21.62% yield). 1 H NMR (400MHz, DMSO-d6) δ10.99(s,1H),8.86(s,1H),7.65(d,J=2.0Hz,1H),7.61-7.49(m,2H),7.21(d,J=8.4Hz,1H),7.15 (dd,J=8.3,2.2Hz,1H),6.88(t,J=6.0Hz,1H),5.11(dd,J=13.3,5.1Hz,1H),4.57(d,J=17.3Hz,1H),4.49-4.34(m,3H),3. 54(t,J=7.0Hz,2H),3.43(t,J=5.8Hz,3H),3.30-3.24(m,2H),3.21-3.14(m,1H),3.05-2.88(m,2H),2.87-2.77(m,3H),2 .67-2.56(m,3H),2.42(td,J=13.3,4.5Hz,1H),2.21(dt,J=12.9,6.4Hz,1H),2.05-1.97(m,1H),1.95-1.86(m,1H); LC-MS m / z(ESI):650.2[M+1]+

[0488] Example 39: Preparation of compound 39

[0489] Following the preparation method of Example 27, the starting material methylamine was replaced with 1,1-dioxide-3-aminothiohexacyclobutane to obtain the title product compound 39 (28 mg, 39.16% yield). 1 H NMR (400MHz, DMSO-d6) δ10.95(s,1H),8.53(s,1H),7.67(d,J=8.0Hz,1H),7.59(s,1H),7.52(s,1H), 7.45(d,J=7.8Hz,1H),7.17(d,J=8.0Hz,1H),7.08(s,1H),6.78(s,1H),5.05(s,1H),4.87(s,1H),4. 42(d,J=17.6Hz,1H),4.32-4.17(m,3H),3.80(d,J=14.2Hz,2H),3.49(d,J=6.9Hz,3H),3.40(s,2H), 2.92-2.80(m,3H),2.64(s,1H),2.54(s,2H),2.30(s,1H),1.98(s,2H),1.39(d,J=7.0Hz,3H).LC-MS m / z(ESI):632.2[M+1] +

[0490] Example 40: Preparation of Compound 40

[0491] Following the preparation method of Example 27, the starting material methylamine was replaced with (S)-3-aminotetrahydrothiophene 1,1-dioxide to obtain the title product compound 40 (104.12 mg, 10.57% yield). 1H NMR (400MHz, DMSO-d6) δ11.00 (s, 1H), 8.60 (d, J = 3.7Hz, 1H), 7.70 (dd, J = 7. 9,1.4Hz,1H),7.62(d,J=2.1Hz,1H),7.55(s,1H),7.48(d,J=7.9Hz,1H),7.2 0(d,J=8.4Hz,1H),7.09(dd,J=8.4,2.2Hz,1H),6.91-6.84(m,1H),5.11(dd ,J=13.2,5.1Hz,1H),4.90(p,J=7.0Hz,1H),4.45(d,J=17.2Hz,1H),4.31(d, J=17.3Hz,1H),3.52(t,J=7.0Hz,2H),3.42(t,J=5.8Hz,3H),3.29(dd,J=13 .1,6.8Hz,2H),3.21-3.14(m,1H),3.03-2.87(m,2H),2.81(dt,J=13.4,6.9H z,3H),2.66-2.56(m,3H),2.43-2.32(m,1H),2.21(dq,J=12.9,6.4Hz,1H),2 .02-1.95(m,1H),1.90(dq,J=13.0,7.8Hz,1H),1.41(d,J=6.9Hz,3H); LC-MS m / z(ESI): 646.2 [M+1] +

[0492] Example 41: Preparation of compound 41

[0493] Following the preparation method of Example 27, the starting material methylamine was replaced with (R)-3-aminotetrahydrothiophene 1,1-dioxide to obtain the title product compound 41 (102.70 mg, 8.17% yield). 1H NMR (400MHz, DMSO-d6) δ10.99(s,1H),8.58(d,J=3.5Hz,1H),7.70(d,J=7.9Hz ,1H),7.62(d,J=2.1Hz,1H),7.55(s,1H),7.48(d,J=8.0Hz,1H),7.20(d,J=8. 4Hz,1H),7.09(dd,J=8.3,2.2Hz,1H),6.85(dd,J=7.7,2.8Hz,1H),5.11(dd,J =13.3,5.1Hz,1H),4.90(p,J=6.9Hz,1H),4.45(d,J=17.3Hz,1H),4.31(d,J=1 7.3Hz,1H),3.53(t,J=7.0Hz,2H),3.42(t,J=5.7Hz,3H),3.29(dd,J=13.1,6. 8Hz,2H),3.18(dt,J=13.7,7.1Hz,1H),3.04-2.86(m,2H),2.81(dt,J=13.8,7 .0Hz,3H),2.67-2.55(m,3H),2.44-2.32(m,1H),2.22(dq,J=13.0,6.4Hz,1H) ,2.03-1.95(m,1H),1.90(dq,J=13.1,7.8Hz,1H),1.42(d,J=6.9Hz,3H); LC-MS m / z(ESI): 646.2 [M+1] + .

[0494] Example 42: Preparation of compound 42

[0495] Step 1: Add 2-chloro-4-nitrostyrene (2 g, 10.89 mmol), toluene (100 mL), compound 42-1 (21.92 g, 108.94 mmol), and poly(ethoxypyrrolidone)phosphononitrile (369.73 mg, 1.09 mmol) to a reaction flask, purge with argon, and react for 16 hours at 40°C in an oil bath. Add silica gel and evaporate the solvent. Purify by column chromatography (PE / EA, 0 to 30 vol%) to give a yellow oily liquid 42-2 (860 mg, 2.23 mmol, 20.51% yield). LC-MS m / z (ESI): 285.1 [M+1-100] + .

[0496] Step 2: Add 42-2 (0.86 g, 2.23 mmol), water (5 mL), ethanol (5 mL), iron powder (624.02 mg, 11.17 mmol), and ammonium chloride (597.65 mg, 11.17 mmol) to the reaction flask, replace with argon gas, and react for 2 hours at 70°C in an oil bath. Recycle the solvent, add water, extract with ethyl acetate, combine the organic phases, dry to anhydrous sodium sulfate, and recycle the solvent again with silica gel. Purify by column chromatography (DCM: CH3OH = 0 vol% to 20 vol%), yielding a yellow oily liquid 42-3 (570 mg, 71.88% yield). LC-MS m / z (ESI): 255.2 [M+1-100] + .

[0497] Step 3: Add 42-3 (130 mg, 366.33 μmol), DCM (2 mL), pyridine (57.95 mg, 732.66 μmol, 59.02 μL), and phenyl chloroformate (68.83 mg, 439.60 μmol) to the reaction flask, and react at 0 °C for 1 hour. Add silica gel to evaporate the solvent, and purify by column chromatography (PE / EA, 0 to 25 vol%) to obtain a yellow oily liquid 42-4 (153 mg, 87.93% yield). LC-MS m / z (ESI): 375.1 [M+1-100] + .

[0498] Step 4: Add 42-4 (153 mg, 322.12 μmol), intermediate 42-6 (92.55 mg, 322.12 μmol), acetonitrile (3 mL), and N,N-diisopropylethylamine (416.32 mg, 3.22 mmol, 561.08 μL) to the reaction flask, purify by purifying with argon gas, and heat in an oil bath to 80 °C for 5 hours. Add silica gel to evaporate the solvent, and purify by column chromatography (DCM / MeOH, 0 to 20 vol%) to obtain a yellow liquid 42-5 (290 mg, 36.67% yield). LC-MS m / z (ESI): 568.3 [M+1-100] + .

[0499] Step 5: Add 42-5 (310 mg, 463.95 μmol), DCM (2 mL), and TFA (1.06 g, 9.28 mmol) to the reaction flask, and react at 20 °C for 2 hours. Recycle to obtain a white solid compound 42 (315 mg, 99.54% yield, TFA). LC-MS m / z (ESI): 568.3 [M+1] + .

[0500] Example A-1: ​​Synthesis of compound DL-1

[0501] Compound 27 (35.00 mg, 64.57 μmol) and compound 1.1A (69.91 mg, 96.86 μmol, CAS No.: 159857-81-5, commercially available) were dissolved in anhydrous DMF (4 mL). Diisopropylethylamine (41.73 mg, 322.86 μmol) was slowly added dropwise to the above reaction system at room temperature, and the reaction was stirred at room temperature for 0.5 hours. The reaction solution was evaporated under reduced pressure to obtain a crude brown oily substance. After acidic separation (acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 10 vol% - 70 vol%), compound DL-1 (26.95 mg, 36.24% yield) was obtained. 1 H NMR (400MHz, DMSO-d6) δ10.96(s,1H),9.96(s,1H),8.53(s,1H),8.06(d,J=7.6Hz,1H),7.78(d,J=8.7Hz,1H),7.68(d,J=7.8Hz ,1H),7.57(dd,J=16.4,8.3Hz,4H),7.47(d,J=8.1Hz,1H),7.26(s,2H),7.11(d,J=29.2Hz,2H),6.98(s,2H),6.80(s,1H),5.95( s,1H),5.39(s,2H),5.08(d,J=13.2Hz,1H),4.92(d,J=28.1Hz,2H),4.59-4.05(m,3H),3.55-3.35(s,15H),3.05-2.73(m,6H), 2.62(d,J=24.5Hz,5H),2.31(s,1H),2.23-1.87(m,3H),1.76-1.29(m,6H),1.17(d,J=7.6Hz,1H),0.81(dd,J=12.4,6.7Hz,6H). LC-MS m / z(ESI):1140.5[M+1] + .

[0502] Example A-2: Synthesis of compound DL-2

[0503] Following the preparation method of Example A-1, compound 27 was replaced with compound 26 to obtain the title product compound DL-2 (35 mg, 30.9% yield). 1H NMR (400MHz, DMSO-d6) δ11.00(s,1H),9.99(s,1H),8.83(s,1H),8.08(d,J=8Hz,1H),7.90(d,J=8Hz,1H),7.66~7.51(m,4H),7.33~7 .27(m,2H),7.21-7.13(m,2H),6.99(s,2H),6.93~6.85(m,2H),5.97(m,1H),5.40(s,2H),5.14~5.00(m,3H),4.56(d,J=20Hz,1H),4 .46~4.35(m,3H),4.19(t,J=8Hz,1H),3.59~3.49(m,5H),3.40~3.34(m,8H),3.22(s,2H),3.04~2.82(m,5H),2.62(t,J=16Hz,1H),2 .46~2.32(m,1H),2.22~2.07(m,2H),2.02~1.93(m,2H),1.68~1.54(m,2H),1.51~1.35(m,6H),1.23~1.14(m,2H),0.86~0.80(m,6H). LC-MS m / z(ESI):1237.4[M+1] + .

[0504] Example A-3: Synthesis of compound DL-3

[0505] Following the preparation method of Example A-1, compound 27 was replaced with compound 9 to obtain the title product compound DL-3 (21 mg, 18.4% yield). 11H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 10.00 (s, 1H), 9.97 (s, 1H), 8.83 (s, 1H), 8.06 (d, J = 7.5 Hz, 1H), 8.03 (d, J = 8.9 Hz, 1H), 7.78 (d, J = 8.6 Hz, 1H), 7.63 (d, J = 2.0 Hz, 1H), 7.61 - 7.54 (m, 3H), 7.51 (t, J = 7.0 Hz, 1H), 7.28 (dd, J = 14.0, 8.0 Hz, 2H), 7.14 (d, J = 16.6 Hz, 1H), 6.97 (s, 2H), 6.84 (s, 1H), 6.73 (d, J = 8.6 Hz, 1H), 5.96 (t, J = 5.8 Hz, 1H), 5.39 (s, 2H), 5.09 (dd, J = 13.3, 5.0 Hz, 1H), 5.02 (d, J = 22.3 Hz, 2H), 4.55 (d, J = 17.3 Hz, 1H), 4.42 (d, J = 5.9 Hz, 1H), 4.40 - 4.30 (m, 2H), 4.16 (t, J = 7.7 Hz, 1H), 3.64 - 3.56 (m, 2H), 3.55 - 3.48 (m, 4H), 2.93 (d, J = 25.9 Hz, 7H), 2.80 (s, 2H), 2.58 (d, J = 16.8 Hz, 1H), 2.63 - 2.54 (m, 2H), 2.11 (ddt, J = 21.0, 13.9, 6.8 Hz, 3H), 1.94 (dd, J = 14.3, 7.5 Hz, 2H), 1.67 (s, 1H), 1.57 (d, J = 9.6 Hz, 1H), 1.45 (p, J = 7.4 Hz, 5H), 1.40 - 1.26 (m, 2H), 1.18 (dt, J = 15.3, 7.8 Hz, 3H), 0.81 (dd, J = 12.4, 6.7 Hz, 6H). LC-MS m / z (ESI): 1236.4 [M+1] + .

[0506] Example A-4: Synthesis of Compound DL-4

[0507] Step 1: Compound 1.4B (2.34 g, 9.58 mmol) was dissolved in DMF (40.0 mL), stirred at room temperature, and diisopropylethylamine (3.71 g, 28.74 mmol, 5.01 mL) was added. Then, a solution of compound 1.4A (2.95 g, 9.58 mmol) dissolved in DMF (40.0 mL) was added. The mixture was heated in an oil bath at 60 °C with stirring for 12 hours. The reaction solution was evaporated to dryness under reduced pressure. The crude product was purified by CombiFlash silica gel column chromatography (40 g, 0–8.0% MeOH / DCM) to give a white solid compound 1.4C (3.91 g, 93.30% yield). LC-MS m / z (ESI): 338.2 [M+1-100] + .

[0508] Step 2: Compounds 1.4C (1.65 g, 3.77 mmol) and HATU (1.72 g, 4.53 mmol) were dissolved in DMF (10.0 mL), stirred at 20 °C, and 1.4D (1.54 g, 3.77 mmol) was added, followed by DIPEA- (1.46 g, 11.31 mmol, 1.97 mL). The reaction mixture was stirred for another hour. The reaction mixture was poured into a separatory funnel, and water (200 mL) was added. Extraction was performed using dichloromethane (50 mL × 2). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and evaporated to dryness under reduced pressure. The crude product was purified by Combi Flash silica gel column chromatography (40 g, 0–3.5% MeOH / DCM) to obtain a yellow oily liquid 1.4E (2.13 g, 57.25% yield). LC-MS m / z (ESI): 849.5 [M + Na] + .

[0509] Step 3: Compound 1.4E (828 mg, 840.92 μmol) was dissolved in THF (4.0 mL), stirred at 20 °C, and pyridine hydrogen fluoride (595.29 mg, 4.20 mmol, 0.60 mL, 70% purity) was added. The reaction mixture was stirred for another 15 minutes. Triethylamine was added to the reaction solution for basification, and the solution was evaporated to dryness under reduced pressure. The crude product was purified by Combi Flash silica gel column chromatography (12 g, 0–5.0% MeOH / DCM) to obtain compound 1.4F (422 mg, 70.40% yield). LC-MS m / z (ESI): 735.4 [M+Na] + .

[0510] Step 4: Compound 1.4F (502 mg, 704.23 μmol) and compound 1.4G (321.35 mg, 1.06 mmol) were dissolved in DCM (5.0 mL), stirred at room temperature, and then DIPEA (364.07 mg, 2.82 mmol, 490.66 μL) was added. The reaction mixture was stirred for another 2.5 hours. The reaction solution was evaporated to dryness under reduced pressure. The crude product was purified by Combi Flash silica gel column chromatography (12 g, 0–5.0% MeOH / DCM) to give compound 1.4H (353 mg, 57.09% yield). LC-MS m / z (ESI): 900.4 [M+Na] + .

[0511] Step 5: Compound 1.4H (353 mg, 402.08 μmol) and compound 7 (230.00 mg, 402.08 μmol) were dissolved in DMF (3.0 mL), stirred at 20 °C, and then DIPEA (519.65 mg, 4.02 mmol, 700.34 μL) was added. The reaction mixture was stirred for another 14 hours. The reaction solution was evaporated to dryness under reduced pressure. The crude product was purified by Combi Flash silica gel column chromatography (12 g, 0–6.3% MeOH / DCM) to give compound 1.4J (289 mg, 54.83% yield). LC-MS m / z (ESI): 1310.5 [M+1] + .

[0512] Step Six: Compound 1.4J (60 mg, 45.77 μmol) was dissolved in DCM (1.8 mL) and stirred at 20 °C. Trifluoroacetic acid (52.19 mg, 457.72 μmol) was then added, and the reaction mixture was stirred for another 30 min. The reaction solution was evaporated to dryness under reduced pressure. DCM (10 mL) and DIPEA were added to fully dissolve and alkalize the solution, and the mixture was evaporated to dryness under reduced pressure again. The crude product was then dried under reduced pressure using an oil pump to obtain compound 1.4K (56 mg, 83.87% yield). LC-MS m / z (ESI): 1210.4 [M+1] + .

[0513] Step 7: Compound 1.4K (28 mg, 19.19 μmol) and HATU (8.76 mg, 23.03 μmol) were dissolved in DMF (1.0 mL) and stirred at 20 °C. Compound 1.4L (8.71 mg, 21.11 μmol) was added, followed by DIPEA (24.81 mg, 191.95 μmol, 33.43 μL). The reaction mixture was stirred for another hour. The reaction solution was evaporated to dryness under reduced pressure. The crude product was purified by HPLC (acid method, GILSON, Waters-Sun Fire C18 column; ACN / Water-0.04% FA; 5%-90%) to obtain compound DL-4 (5.04 mg, 14.46% yield). 1 H NMR(400MHz,MeOD)δ7.62(dt,J=13.1,7.8Hz,4H),7.53(s,1H),7.30(d,J=6.8Hz, 2H),7.09(s,2H),6.76(d,J=2.1Hz,2H),5.14(dd,J=13.3,5.1Hz,1H),5.05(s,2H ),4.64-4.48(m,5H),4.43(q,J=7.1Hz,1H),4.18-3.98(m,2H),3.85(d,J=13.4Hz ,1H),3.77-3.69(m,2H),3.69-3.49(m,28H),3.44(q,J=6.4,5.4Hz,5H),3.34(d, J=1.0Hz,3H),2.96-2.82(m,3H),2.78(d,J=17.3Hz,1H),2.74-2.55(m,2H),2.54 -2.44(m,2H),2.31-2.22(m,2H),2.22-2.12(m,3H),2.09(d,J=6.2Hz,1H),2.02( s,3H),1.57(dp,J=22.5,7.3Hz,5H),1.46(dd,J=7.5,4.4Hz,3H),1.31(d,J=16.3 Hz, 5H), 0.97 (dd, J = 9.5, 6.9Hz, 6H), 0.70 (d, J = 6.7Hz, 2H), 0.60 (d, J = 3.6Hz, 2H). LC-MS m / z(ESI):825.0[M / 2+23] + .

[0514] Example A-5: Synthesis of compound DL-5

[0515] Following the preparation method of Example A-4, 1.4 L of the starting material was replaced with 1.5 A to obtain the title product compound DL-5 (5 mg, 13.9% yield).1 H NMR(400MHz,MeOD)δ8.51(s,1H),7.62(dt,J=12.5,7.8Hz,4H),7.53(s,1H),7.31 (dd,J=5.6,3.6Hz,2H),7.09(s,2H),6.76(d,J=2.3Hz,2H),5.17(d,J=5.1Hz,1H) ,5.13(d,J=5.1Hz,1H),5.05(s,2H),4.59(s,6H),4.58-4.52(m,4H),4.18-3.99( m,2H),3.72(t,J=5.4Hz,2H),3.69-3.54(m,32H),3.53-3.51(m,2H),3.44(q,J=6. 4,5.3Hz,6H),3.34(s,3H),2.86(t,J=6.9Hz,2H),2.82-2.75(m,1H),2.72-2.57( m,2H),2.51(dd,J=13.1,4.9Hz,2H),2.30-2.24(m,2H),2.21-2.16(m,2H),1.63-1 .57(m,2H),1.57-1.51(m,2H),1.46(dd,J=7.2,4.4Hz,3H),1.30(dd,J=13.5,5.7 Hz, 4H), 0.97 (dd, J = 8.4, 5.7Hz, 6H), 0.70 (d, J = 6.9Hz, 2H), 0.60 (d, J = 3.8Hz, 2H). LC-MS m / z(ESI):847.0[M / 2+23] + .

[0516] Example A-6: Synthesis of compound DL-6

[0517] Following the preparation method of Example A-4, 1.4 L of the starting material was replaced with 1.6 A to obtain the title product compound DL-6 (10 mg, 19.8% yield). 1H NMR (400MHz, DMSO-d6) δ10.99(s,1H),9.76(s,1H),8.79(s,1H),8.09(s,1H),8.01(s,1H),7.63(d,J=2.1 Hz,1H),7.58(dd,J=11.5,5.0Hz,3H),7.54-7.48(m,1H),7.35(s,1H),7.27(d,J=8.1Hz,2H),7.16(d,J=8 .4Hz,1H),7.11(dd,J=8.3,2.1Hz,1H),6.96(s,2H),6.80(t,J=6.0Hz,1H),5.10(dd,J=13.3,5.1Hz,1H), 4.98(s,2H),4.55(d,J=17.4Hz,1H),4.42(d,J=6.0Hz,2H),4.37(d,J=17.4Hz,2H),4.32(s,3H),4.28-4. 09(m,6H),4.09-3.83(m,6H),3.57(s,1H),3.49(dt,J=13.0,6.2Hz,6H),3.20(s,1H),3.00-2.83(m,11H) ,2.83-2.64(m,12H),2.58(d,J=17.5Hz,1H),2.45-2.35(m,1H),2.15(d,J=7.6Hz,2H),2.08-1.89(m,6H) ,1.82(s,1H),1.76(dd,J=17.7,8.8Hz,2H),1.45(d,J=14.7Hz,4H),1.29(d,J=7.0Hz,3H),1.21(d,J=10. 9Hz, 3H), 0.84 (d, J = 6.7Hz, 3H), 0.77 (d, J = 6.7Hz, 3H), 0.63 (dd, J = 7.0, 5.0Hz, 2H), 0.55 (d, J = 3.4Hz, 2H). LC-MS m / z(ESI):840.5[M / 2+1] + .

[0518] Example A-7: Synthesis of compound DL-7

[0519] Following the preparation method of Example A-4, the title product compound DL-7 (9.6 mg, 13.4% yield) was obtained by replacing 1.4 L of the starting material with 1.7 A. 1H NMR (400MHz, DMSO-d6) δ10.98(s,1H),9.71(d,J=5.4Hz,1H),8.77(s,1H),8.06(d,J=7.0Hz,1H),7.99(s,1H),7.6 5-7.54(m,4H),7.54-7.48(m,1H),7.39-7.32(m,1H),7.27(d,J=8.2Hz,2H),7.21-7.08(m,2H),6.95(s,2H),6.79( t,J=6.0Hz,1H),5.09(dd,J=13.3,5.1Hz,1H),4.98(s,2H),4.55(d,J=17.4Hz,1H),4.42(d,J=5.9Hz,2H),4.38(d ,J=17.3Hz,1H),4.32(t,J=7.1Hz,1H),4.10(dd,J=7.8,5.5Hz,1H),4.01(s,1H),3.68(d,J=14.0Hz,1H),3.59(t,J =6.8Hz,2H),3.52(d,J=6.9Hz,2H),3.48(d,J=1.4Hz,48H),3.42-3.39(m,3H),3.32-3.27(m,3H),3.22(s,3H),2. 89(ddd,J=17.4,13.5,5.4Hz,2H),2.79(t,J=6.9Hz,2H),2.64-2.51(m,3H),2.45-2.36(m,1H),2.16(t,J=7.3Hz,2 H),2.07-1.96(m,3H),1.88(t,J=16.0Hz,2H),1.70(d,J=11.6Hz,1H),1.52-1.41(m,4H),1.29(d,J=7.1Hz,3H),1 .26-1.11(m,4H),0.84(d,J=6.8Hz,3H),0.78(d,J=6.7Hz,3H),0.63(dd,J=7.0,4.8Hz,2H),0.55(d,J=4.0Hz,2H). LC-MS m / z(ESI):935.50[M / 2+23] + .

[0520] Example A-8: Synthesis of compound DL-8

[0521] Step 1: Compound 1.4K (105 mg, 60.71 μmol) and HATU (27.70 mg, 72.85 μmol) were dissolved in DMF (1.5 mL), stirred at 20 °C, and compound 1.8A (53.00 mg, 61.78 μmol) was added, followed by DIPEA (78.46 mg, 607.07 μmol, 105.74 μL). The reaction mixture was stirred for another hour. The reaction solution was evaporated to dryness under reduced pressure. The crude product was purified by Combi Flash silica gel column chromatography (4 g, 0–11.0% MeOH / DCM) to obtain compound 1.8B (84 mg, 16.87% yield). LC-MS m / z (ESI): 1025.5 [M / 2+1] + .

[0522] Step 2: Compound 1.8B (74.00 mg, 9.02 μmol) was placed in a reaction flask, and a solution of trifluoroacetic acid (20.57 mg, 180.43 μmol, 0.10 mL) dissolved in DCM (0.5 mL) was added. The mixture was stirred at 20 °C for 20 min. The reaction solution was then evaporated to dryness under reduced pressure at 40 °C. The crude product was purified by HPLC (acid method, GILSON, Waters-Sun Fire C18 column; ACN / Water-0.04% FA; 5%-90%) to obtain compound DL-8 (4.67 mg, 24.84% yield). 1H NMR (400MHz, DMSO-d6) δ10.97(s,1H),9.73(s,1H),8.78(s,1H),8.06(d,J=7.0Hz,1H),7.99(s,1H),7.64-7. 59(m,2H),7.56(d,J=7.8Hz,2H),7.51(t,J=7.0Hz,1H),7.35(d,J=8.0Hz,2H),7.27(d,J=8.1Hz,2H),7.16(d ,J=3.6Hz,1H),7.16-7.09(m,2H),6.95(s,2H),6.80(s,1H),5.39(s,1H),5.09(dd,J=13.2,5.0Hz,1H),4.98 (s,2H),4.75(s,1H),4.57(s,3H),4.50(s,1H),4.43(d,J=6.1Hz,2H),4.38(d,J=17.6Hz,2H),4.32(d,J=7.0H z,1H),4.13-4.09(m,1H),4.01(s,1H),3.97(s,3H),3.76(s,2H),3.66(s,4H),3.59(t,J=6.1Hz,6H),3.55-3 .45(m,38H),2.90(s,2H),2.81(d,J=7.0Hz,2H),2.69(s,1H),2.59(d,J=18.1Hz,3H),2.41(s,1H),2.16(t,J= 7.5Hz,2H),1.99(d,J=16.1Hz,3H),1.89(s,2H),1.71(s,1H),1.44(d,J=12.5Hz,4H),1.29(d,J=7.0Hz,3H), 1.25-1.14(m,3H),0.84(d,J=6.7Hz,3H),0.78(d,J=6.8Hz,3H),0.64(d,J=6.7Hz,2H),0.55(d,J=3.6Hz,2H). LC-MS m / z(ESI):937.4[M / 2+1] + .

[0523] Example A-9: Synthesis of compound DL-9

[0524] Following the preparation method of Example A-4, compound 7 was replaced with compound 9, and 1.4 L of starting material was replaced with 1.9 D, to obtain the title product compound DL-9 (25 mg, 33.6% yield). 1H NMR (400MHz, DMSO-d6) δ10.98(s,1H),9.74(s,1H),8.77(s,1H),8.07(d,J=7.0Hz,1H),7.98(s,1H),7.6 3(d,J=2.1Hz,1H),7.62-7.54(m,3H),7.54-7.48(m,1H),7.39-7.23(m,3H),7.14(d,J=18.1Hz,2H),6.96 (s,2H),6.79(t,J=6.0Hz,1H),5.10(dd,J=13.2,5.1Hz,1H),5.00(s,2H),4.55(d,J=17.4Hz,1H),4.43(d ,J=6.0Hz,2H),4.37(d,J=17.3Hz,1H),4.33(t,J=7.0Hz,1H),4.14-4.08(m,1H),4.01(s,1H),3.68(d,J= 14.2Hz,1H),3.59(t,J=6.1Hz,4H),3.54-3.44(m,48H),3.43-3.39(m,3H),3.37(d,J=7.2Hz,2H),3.34(s ,4H),3.22(s,3H),2.96(s,2H),2.90(s,2H),2.81(d,J=7.9Hz,2H),2.57(t,J=14.2Hz,3H),2.45-2.35(m ,1H),2.16(t,J=7.4Hz,2H),2.06(s,3H),2.04-1.94(m,3H),1.94-1.78(m,2H),1.71(s,1H),1.45(dq,J= 15.0, 7.3Hz, 4H), 1.29 (d, J = 7.1Hz, 3H), 1.24-1.14 (m, 2H), 0.84 (d, J = 6.7Hz, 3H), 0.78 (d, J = 6.8Hz, 3H). LC-MS m / z(ESI):946.5[M / 2+1] + .

[0525] Example A-10: Synthesis of compound DL-10

[0526] Following the preparation method of Examples A-8, 1.8A was replaced with 1.10A and 1.4K was replaced with 1.9C to obtain the title product compound DL-10 (5.6 mg, 7.8% yield). 1H NMR (400MHz, DMSO-d6) δ10.98(s,1H),9.74(s,1H),8.80(s,1H),8.07(d,J=7.2Hz,2H),8.00(s,1H),7. 63(d,J=2.1Hz,1H),7.62-7.54(m,3H),7.54-7.48(m,1H),7.32(dt,J=21.7,8.5Hz,4H),7.21(d,J=7.9 Hz,1H),7.14(d,J=16.3Hz,2H),6.96(d,J=3.8Hz,2H),6.82(t,J=5.9Hz,1H),5.40(s,2H),5.09(dd,J= 13.3,5.1Hz,1H),5.00(s,2H),4.77(s,2H),4.60-4.49(m,5H),4.46-4.38(m,5H),4.33(dd,J=12.9,5.9 Hz,2H),4.11(t,J=6.7Hz,1H),3.98(s,3H),3.76(s,2H),3.66(s,4H),3.61-3.54(m,10H),3.48(d,J=5 .8Hz,54H),2.96(s,2H),2.90(s,2H),2.81(d,J=8.1Hz,2H),2.59-2.53(m,3H),2.42(dd,J=13.0,4.5H z,1H),2.16(t,J=7.1Hz,2H),2.06(s,3H),1.99(d,J=15.3Hz,3H),1.90(d,J=13.7Hz,2H),1.71(s,1H) ,1.46(s,4H),1.29(d,J=6.9Hz,3H),1.26-1.16(m,4H),0.84(dd,J=7.0,2.7Hz,3H),0.79-0.76(m,3H). LC-MS m / z(ESI):1102.6[M / 2+1] + .

[0527] Example A-11 Synthesis of compound DL-11

[0528] Following the preparation method of Examples A-8, the starting material 1.8A was replaced with 1.10A to obtain the title product compound DL-11 (1.2 mg, 35.6% yield). 11H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 9.71 (s, 1H), 8.77 (s, 1H), 8.05 (d, J = 7.1 Hz, 1H), 7.99 (s, 1H), 7.63 (d, J = 7.5 Hz, 2H), 7.60 - 7.55 (m, 2H), 7.52 (d, J = 6.5 Hz, 1H), 7.41 (d, J = 8.4 Hz, 1H), 7.33 (d, J = 5.9 Hz, 2H), 7.27 (d, J = 8.2 Hz, 2H), 7.12 (d, J = 2.0 Hz, 2H), 6.95 (s, 2H), 6.79 (s, 1H), 5.11 (s, 1H), 5.07 (dd, J = 13.2, 5.0 Hz, 1H), 5.02 (s, 1H), 4.98 (s, 2H), 4.57 (s, 1H), 4.53 (s, 1H), 4.43 (d, J = 6.0 Hz, 2H), 4.40 (s, 2H), 4.36 (s, 1H), 4.33 (s, 2H), 4.12 - 4.08 (m, 2H), 4.02 - 3.99 (m, 2H), 3.98 - 3.95 (m, 2H), 3.57 (s, 4H), 3.53 (s, 6H), 3.48 (d, J = 3.0 Hz, 47H), 3.44 (s, 4H), 3.43 (s, 4H), 3.40 (s, 4H), 3.38 (s, 4H), 2.79 (d, J = 6.5 Hz, 1H), 2.74 - 2.67 (m, 1H), 2.67 - 2.63 (m, 2H), 2.62 - 2.58 (m, 1H), 2.56 (s, 3H), 2.47 - 2.43 (m, 1H), 2.43 - 2.39 (m, 1H), 2.38 (s, 1H), 2.37 - 2.32 (m, 1H), 2.32 - 2.29 (m, 1H), 2.22 - 2.17 (m, 1H), 2.15 (d, J = 7.5 Hz, 1H), 2.01 (s, 1H), 1.94 - 1.81 (m, 1H), 1.46 (s, 4H), 1.29 (d, J = 7.1 Hz, 3H), 1.20 (d, J = 14.0 Hz, 3H), 0.84 (d, J = 6.6 Hz, 3H), 0.78 (d, J = 6.8 Hz, 3H), 0.64 (d, J = 7.0 Hz, 2H), 0.55 (d, J = 3.9 Hz, 2H). LC-MS m / z (ESI): 1091.6 [M / 2 + 23] + .

[0529] Example A-12: Synthesis of Compound DL-12

[0530] Following the preparation method of Examples A-8, the starting material 1.8A was replaced with 1.12A to obtain the title product compound DL-12 (6 mg, 14.2% yield). 1 H NMR (400MHz, DMSO-d6) δ10.98(s,1H),9.76(s,1H),8.81(s,1H),8.08(d,J=7.4Hz,2H),8.00(s,1H),7.63(d,J=2. 2Hz,1H),7.62-7.58(m,1H),7.56(d,J=7.5Hz,2H),7.52(d,J=6.8Hz,1H),7.34(s,3H),7.28(d,J=8.3Hz,1H),7.21 (d,J=8.0Hz,1H),7.19-7.09(m,2H),6.96(s,2H),6.82(s,1H),5.40(s,1H),5.10(dd,J=13.4,5.1Hz,1H),5.02(d, J=24.8Hz,2H),4.76(s,1H),4.55(d,J=17.1Hz,3H),4.51(s,1H),4.43(d,J=6.2Hz,4H),4.38(d,J=17.8Hz,2H),4. 32(d,J=7.1Hz,1H),4.12(s,1H),3.99(s,3H),3.76(s,2H),3.67(d,J=7.8Hz,4H),3.59(d,J=6.6Hz,6H),3.48(d,J =11.0Hz,24H),2.95-2.85(m,2H),2.80(t,J=6.9Hz,2H),2.68(s,1H),2.59(d,J=11.5Hz,3H),2.42(dd,J=12.9,4. 4Hz,1H),2.16(s,2H),1.99(d,J=14.6Hz,3H),1.89(s,2H),1.70(s,1H),1.46(s,4H),1.29(d,J=7.0Hz,3H),1.21( dd,J=17.5,7.5Hz,3H),0.84(d,J=6.6Hz,3H),0.78(d,J=6.3Hz,3H),0.64(d,J=7.0Hz,2H),0.55(d,J=3.9Hz,2H). LC-MS m / z(ESI):872.0[M / 2+1] + .

[0531] Example A-13: Synthesis of compound DL-13

[0532] Step 1: Compound 1.13B (0.6 g, 1.47 mmol) and compound 1.13A (392.67 mg, 1.91 mmol) were dissolved in a mixture of dichloromethane (30 mL) and methanol (2 mL). The reaction mixture was kept clear. After the reaction system was cooled to 0 °C, EEDQ (473.22 mg, 1.91 mmol) was added, and the reaction was stirred for 18 hours. The reaction mixture was concentrated to obtain a brown oil. The brown oil was subjected to silica gel column chromatography (DCM:MeOH = 20:1) to obtain compound 1.13C (0.73 g, 83.38% yield); LC-MS m / z (ESI): 617.4 [M+23] + .

[0533] Step 2: Compound 1.13C (600 mg, 1.01 mmol) was dissolved in DCM (15 mL). After the reaction temperature was lowered to 0 °C, chlorosulfonic acid isocyanate (171.32 mg, 1.21 mmol) was added, followed by triethylamine (408.29 mg, 4.03 mmol, 562.77 μL). Finally, a solution of 1.13I (386.82 mg, 1.01 mmol) in dichloromethane (5 mL) was slowly added dropwise to the above reaction system. The reaction was stirred at room temperature for 0.5 hours. The reaction solution was concentrated and subjected to silica gel column chromatography (DCM:MeOH = 20:1) to obtain compound 1.13D (2 g, 1.85 mmol, 91.51% yield). LC-MS m / z (ESI): 1105.5 [M+23] + .

[0534] Step 3: Compound 1.13D (1.8 g, 1.66 mmol) was dissolved in THF (10 mL) and added to TBAF (1 M, 10 mL). The reaction mixture was stirred at room temperature for 2 hours. After the organic solvent was evaporated, 100 mL of dichloromethane and 60 mL of water were added to the mixture to separate the organic phase. The organic phase was washed repeatedly with water 6 times and with sodium chloride aqueous solution 2 times in a separatory funnel. The organic phases were then combined, dried over sodium sulfate, and concentrated to obtain a brown oily substance. After purification by silica gel column chromatography (DCM:MeOH = 10:1), compound 1.13E (1.4 g, 86.95% yield) was obtained. LC-MS m / z (ESI): 991.4 [M+23] + .

[0535] Step 4: Compound 1.13E (300 mg, 309.56 μmol) was dissolved in DCM (6 mL), DIPEA (120.02 mg, 928.69 μmol, 161.76 μL) was added, and finally di(p-nitrobenzene) carbonate (98.88 mg, 325.04 μmol) was added. The reaction mixture was stirred gradually at room temperature for 18 hours. The reaction solution was concentrated and then subjected to silica gel column chromatography (DCM:MeOH = 20:1) to give compound 1.13F (190 mg, 54.11% yield). LC-MS m / z (ESI): 1156.50 [M+23] + .

[0536] Step 5: Compound 7 (15.13 mg, 26.45 μmol) and compound 1.13F (25 mg, 22.04 μmol) were dissolved in DMF (1 mL), ensuring the reaction system remained clear. Following this, DIPEA (85.46 mg, 661.26 μmol, 115.18 μL) was added, and the reaction was stirred at room temperature for 48 hours. After concentration, the mixture was subjected to silica gel column chromatography (DCM:MeOH = 10:1) to obtain compound 1.13G (34 mg, 98.43% yield). LC-MS m / z (ESI): 734.0 [(M-100) / 2+1] + .

[0537] Step Six: Dissolve compound 1.13G (35 mg, 22.33 μmol) in DCM (2 mL), add TFA (0.5 mL), and stir at room temperature for 2 hours. After concentration, a brown oily crude product is obtained. After rotary evaporation to dryness, compound 1.13H (32 mg, crude product) is obtained, which is used directly in the next reaction. LC-MS m / z (ESI): 734.0 [M / 2+1] + .

[0538] Step 7: Compound 1.13H (35 mg, 23.86 μmol) was dissolved in DMF (2 mL) to ensure the reaction system remained clear. Then, DIPEA (18.50 mg, 143.15 μmol, 24.93 μL) was added. Finally, 6-(maleimide)hexanoic acid succinimide ester (9.56 mg, 31.02 μmol) dissolved in DMF (0.5 mL) was added dropwise. The reaction was stirred at room temperature for 18 hours. After concentration, a brown oil was obtained. The brown oil was subjected to acidic (acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 15%-65%) reversed-phase separation, followed by freeze-drying to obtain compound DL-13 (8.95 mg, 22.60% yield). 1 H NMR(400MHz, CDCl3)δ9.50(s,1H),9.34-9.05(m,2H),7.77(s,2H),7.62-7.38(m,5H) ,7.24(s,2H),6.91(s,2H),6.74(s,1H),6.67(s,2H),6.34(s,1H),5.18(s,1H),4.99( d,J=31.1Hz,3H),4.76-4.17(m,7H),3.87-3.28(m,34H),3.22(s,2H),2.81(s,3H),2 .14(d,J=89.7Hz,17H),1.56(t,J=17.0Hz,3H),1.47-1.14(m,5H),1.01-0.45(m,9H). LC-MS m / z(ESI):830.3[M / 2+1] + .

[0539] Example A-14: Synthesis of compound DL-14

[0540] Following the preparation method of Example A-13, compound 7 was replaced with compound 19 to obtain the title product compound DL-14 (35 mg, 25.56% yield). 1H NMR(400MHz,DMSO-d6)δ11.17(s,1H),11.00(s,1H),9.98(s,1H),8.79(s,1H),8.21(dd, J=26.4,7.3Hz,2H),7.81(d,J=8.7Hz,1H),7.70-7.46(m,6H),7.28(d,J=8.3Hz,2H),7.1 4(s,2H),6.99(s,2H),6.81(t,J=6.0Hz,1H),5.11(dd,J=13.3,5.1Hz,1H),4.98(s,2H), 4.70-4.60(m,1H),4.56(d,J=17.4Hz,1H),4.48-4.34(m,4H),4.30(dd,J=11.0,4.4Hz,1H ),4.22(dd,J=8.7,6.3Hz,1H),4.10(dd,J=10.9,8.1Hz,1H),3.68-3.34(m,39H),3.23(s ,3H),3.04(q,J=6.0Hz,2H),2.97-2.85(m,1H),2.79(d,J=3.9Hz,1H),2.60(d,J=17.2Hz, 1H),2.43(dd,J=13.1,4.5Hz,1H),2.11(t,J=7.4Hz,2H),2.06-1.91(m,1H),1.87-1.58( m, 3H), 1.48 (h, J = 7.6Hz, 5H), 1.25 (dd, J = 34.9, 7.1Hz, 6H), 0.83 (dd, J = 18.8, 6.7Hz, 7H). LC-MS m / z(ESI):830.4[M / 2+1] + .

[0541] Example A-15: Synthesis of compound DL-15

[0542] Following the preparation method of Example A-13, compound 7 was replaced with compound 16 to obtain the title product compound DL-15 (20.5 mg, 18.2% yield). 1H NMR(400MHz,DMSO-d6)δ11.15(s,1H),10.97(s,1H),9.96(s,1H),8.78(s,1H) ,8.18(dd,J=25.0,7.4Hz,2H),7.79(d,J=8.7Hz,1H),7.67-7.45(m,5H),7.33 -7.04(m,4H),6.97(d,J=2.3Hz,2H),6.79(t,J=6.0Hz,1H),5.09(dd,J=13.3, 5.0Hz,1H),5.02-4.87(m,2H),4.62(td,J=7.8,4.4Hz,1H),4.54(d,J=17.4Hz ,1H),4.49-4.31(m,4H),4.24(ddd,J=31.1,9.9,5.4Hz,2H),4.13-3.98(m,2H ),3.62-3.30(m,41H),3.21(d,J=1.4Hz,4H),3.02(q,J=6.0Hz,2H),2.92-2.7 2(m,2H),2.60(s,1H),2.09(t,J=7.5Hz,2H),1.97(d,J=6.8Hz,1H),1.45(q,J =8.8,7.6Hz,4H), 1.23(dd,J=35.6,7.2Hz,6H), 0.81(dd,J=18.8,6.7Hz,7H). LC-MS m / z(ESI): 846.0 [M / 2+23] + .

[0543] Example A-16: Synthesis of compound DL-16

[0544] Following the preparation method of Example A-13, the starting material 1.13I was replaced with 1.16A to obtain the title product compound DL-16 (10 mg, 18.1% yield). 1H NMR (400MHz, DMSO-d6) δ11.15(s,1H),10.97(s,1H),9.96(s,1H),8.77(s,1H),8.19(dd,J=26.0,7.3Hz,2H),7.78(d,J=8.8Hz,1H),7.66-7.45(m ,5H),7.26(d,J=8.2Hz,2H),7.19-7.06(m,2H),6.96(s,2H),6.78(t,J= 6.0Hz,1H),5.09(dd,J=13.3,5.1Hz,1H),4.97(s,2H),4.68-4.47(m,2H) ,4.49-3.94(m,6H),3.57-3.31(m,51H),3.21(s,3H),3.11-2.72(m,5H) ,2.58(d,J=17.0Hz,1H),2.45-2.28(m,1H),2.09(t,J=7.5Hz,2H),1.96( dd,J=12.9,7.0Hz,3H),1.44(dt,J=15.3,8.5Hz,5H),1.27(d,J=7.0Hz,3H),1.24-1.09(m,2H),0.81(dd,J=19.0,6.7Hz,6H),0.70-0.45(m,4H). LC-MS m / z(ESI): 919.0 [M / 2+23] + .

[0545] Example A-17: Synthesis of compound DL-17

[0546] Following the preparation method of Example A-16, compound 7 was replaced with compound 15 to obtain the title product compound DL-17 (18.8 mg, 15.0% yield). 1H NMR (400MHz, DMSO-d6) δ11.15(s,1H),10.98(s,1H),9.97(s,1H),8.78(s,1H),8.19(dd,J=28.5,7.4Hz,2H),7.79(d,J=8.7Hz,1H),7.70-7.4 1(m,6H),7.35-7.05(m,4H),6.96(s,2H),6.78(t,J=6.0Hz,1H),5.09( dd,J=13.3,5.1Hz,1H),4.96(s,2H),4.67-4.58(m,1H),4.54(d,J=17.4 Hz,1H),4.47-4.31(m,4H),4.24(ddd,J=29.1,9.7,5.3Hz,2H),4.08(dd,J=10.9,8.2Hz,1H),3.61-3.30(m,56H),3.09-2.72(m,6H),2.64-2.5 0(m,2H),2.46-2.12(m,2H),2.09(t,J=7.4Hz,2H),2.03-1.86(m,1H),1.45(h,J=7.6Hz,5H),1.34-1.10(m,6H),0.81(dd,J=19.3,6.7Hz,6H). LC-MS m / z(ESI):950.5[M / 2+23] + .

[0547] Example A-18: Synthesis of compound DL-18

[0548] Following the preparation method of Example A-16, compound 7 was replaced with compound 20 to obtain the title product compound DL-18 (27.8 mg, 23.2% yield). 1H NMR (400MHz, DMSO-d6) δ11.15(s,1H),10.98(s,1H),9.97(s,1H),8.78(s,1H),8.19(dd,J=25.2,7.4Hz,2H),7.79(d,J=8.7Hz,1 H),7.69-7.41(m,5H),7.36-7.04(m,4H),6.96(s,2H),6.79(t,J=6.0Hz,1H),5.08(dt,J=13.1,6.5Hz,1H),4.92(s,2H),4.69-4. 47(m,2H),4.46-3.93(m,9H),3.67(s,2H),3.57-3.31(m,48H),3.21(s,3H),3.11-2.73(m,4H),2.60(t,J=18.2Hz,1H),2.45-2.2 4(m,1H),2.03(dt,J=46.1,7.1Hz,4H),1.43(dd,J=15.7,7.7Hz,4H),1.23(dd,J=35.7,7.3Hz,6H),0.81(dd,J=18.9,6.7Hz,6H). LC-MS m / z(ESI):905.0[M / 2+23] + .

[0549] Example A-19: Synthesis of compound DL-19

[0550] Following the preparation method of Example A-4, starting material 1.4B was replaced with 1.19A and starting material 1.4L was replaced with 1.19H to obtain the title product compound DL-19 (8 mg, 18.8% yield). 1H NMR(400MHz,DMSO-d6)δ10.98(s,1H),9.95(s,1H),8.81(s,1H),8.38-8.23(m,1H),8.03-8.00(m,1H), 7.77-7.74(m,1H),7.66(s,1H),7.60-7.48(m,4H),7.28-7.22(m,2H),7.20-7.05(m,7H),6.94(s,2H),6 .68(s,1H),5.11-5.04(m,1H),4.96(s,2H),4.44-4.15(m,19H),4.11-3.78(m,13H),3.58-3.43(m,5H), 2.98-2.63(m,34H),1.95(s,9H),1.40-1.18(m,8H),0.85-0.77(m,9H),0.65(s,2H),0.55(s,2H).LC-MS m / z(ESI):1006.9[M / 2+1] + .

[0551] Example A-21: Synthesis of compound DL-21

[0552] Following the preparation method of Example A-16, compound 7 was replaced with compound 27 to obtain the title product compound DL-21 (32 mg, 12.77% yield). 1H NMR (400MHz, DMSO-d6) δ11.16(s,1H),10.97(s,1H),9.97(s,1H),8.54(d,J=3.2Hz, 1H),8.21(dd,J=24.7,7.4Hz,2H),7.80(d,J=8.8Hz,1H),7.69(d,J=7.8Hz,1H),7.66 -7.52(m,5H),7.48(d,J=7.9Hz,1H),7.28(d,J=8.0Hz,2H),7.20-7.03(m,1H),6.98 (s,1H),6.84-6.75(m,1H),5.10(dd,J=13.3,5.1Hz,1H),5.02-4.84(m,3H),4.70-4. 59(m,1H),4.51-4.26(m,4H),4.22(dd,J=8.7,6.2Hz,1H),4.11(dd,J=10.9,8.1Hz, 1H),3.61-3.35(m,47H),3.23(s,3H),3.04(q,J=6.0Hz,2H),2.97-2.72(m,7H),2.60 (d,J=16.9Hz,1H),2.46-2.29(m,1H),2.11(t,J=7.4Hz,2H),2.05-1.91(m,3H),1.45 (dd, J = 23.7, 7.0 Hz, 8H), 1.25 (dd, J = 34.3, 7.2 Hz, 6H), 0.83 (dd, J = 18.8, 6.7 Hz, 7H). LC-MS m / z(ESI):903.5[M / 2+23] + .

[0553] Example A-22: Synthesis of compound DL-22

[0554] Following the preparation method of Example A-4, starting compound 7 was replaced with compound 27, and starting material 1.4L was replaced with 1.7A to obtain the title product compound DL-22 (12 mg, 19% yield). 1H NMR (400MHz, DMSO-d6) δ10.95(s,1H),9.72(s,1H),8.52(d,J=3.2Hz,1H),8.06(d,J=7.0Hz,1H),7.98(s,1H),7.67(d,J=7.8Hz,1H),7.63-7 .50(m,4H),7.46(d,J=7.9Hz,1H),7.34(s,1H),7.26(s,1H),7.13(t,J=9.7Hz,1H),7.06(s,1H),6.94(s,2H),6.83-6.73(m,1H),5.13-5.01 (m,1H),4.95(s,2H),4.88(t,J=7.1Hz,1H),4.49-4.20(m,2H),4.14-3.91(m,2H),3.74-3.35(m,68H),3.21(s,3H),3.01-2.69(m,7H),2.69 -2.50(m,2H),2.43-2.25(m,1H),2.15(t,J=7.3Hz,2H),2.08-1.61(m,5H),1.42(dq,J=29.9,9.0,8.2Hz,7H),0.80(dd,J=22.5,6.8Hz,7H). LC-MS m / z(ESI):920.5[M / 2+23] + .

[0555] Example A-23: Synthesis of compound DL-23

[0556] Following the preparation method of Example A-22, starting material 1.7A was replaced with 1.23B to obtain the title product compound DL-23 (12 mg, 19% yield). 1H NMR (400MHz, DMSO-d6) δ10.94(s,1H),9.71(s,1H),8.52(s,1H),8.05(d,J=7.0Hz,1H),7.98(s,1H),7.67(d,J= 7.9Hz,1H),7.62-7.54(m,3H),7.53(s,1H),7.45(d,J=7.9Hz,1H),7.33(s,1H),7.25(d,J=7.8Hz,2H),7.13(s,1 H),7.05(d,J=8.0Hz,1H),6.94(s,2H),6.79(d,J=7.4Hz,1H),5.07(dd,J=13.2,5.1Hz,1H),4.95(s,2H),4.88(t ,J=7.2Hz,1H),4.49(d,J=5.3Hz,2H),4.42(d,J=17.0Hz,2H),4.30(t,J=14.3Hz,5H),4.10(dd,J=7.8,5.5Hz,1H ),4.00(s,1H),3.79(s,1H),3.73-3.63(m,2H),3.59(d,J=6.7Hz,7H),3.48(d,J=16.5Hz,53H),3.39(d,J=7.0Hz ,3H),3.34(d,J=7.6Hz,5H),2.95-2.83(m,3H),2.79(d,J=9.1Hz,7H),2.59(s,1H),2.54(s,2H),2.36(d,J=13.2 Hz,1H),2.15(t,J=7.1Hz,3H),2.06-1.93(m,4H),1.85(d,J=18.4Hz,3H),1.69(s,1H),1.43(d,J=12.4Hz,4H),1 .39(d,J=7.0Hz,4H), 1.28(d,J=7.0Hz,4H), 1.18(d,J=7.6Hz,3H), 0.82(d,J=6.8Hz,3H), 0.77(d,J=6.8Hz,3H). LC-MS m / z(ESI):1054.4[M / 2+1] + .

[0557] Example A-24: Synthesis of compound DL-24

[0558] Following the preparation method of Example A-22, starting material 1.7A was replaced with 1.24B to obtain the title product compound DL-24 (6.8 mg, 9.7% yield). 1H NMR (400MHz, DMSO-d6) δ10.94(s,1H),9.72(s,1H),8.52(d,J=3.1Hz,1H),8.06(d,J=7.3Hz,1H),7.97(s,1H),7 .67(d,J=7.8Hz,1H),7.62-7.54(m,3H),7.51(d,J=13.5Hz,2H),7.45(d,J=7.9Hz,1H),7.33(d,J=7.9Hz,1H),7. 25(s,2H),7.20(d,J=8.0Hz,1H),7.13(s,1H),7.06(s,1H),6.94(s,2H),6.79(d,J=7.3Hz,1H),5.07(dd,J=12.9 ,5.1Hz,1H),4.95(s,2H),4.91-4.83(m,1H),4.44(d,J=4.7Hz,2H),4.39(d,J=4.9Hz,2H),4.29(q,J=8.7,6.3Hz ,5H),4.10(t,J=6.9Hz,1H),4.00(s,1H),3.65(s,2H),3.57(d,J=11.1Hz,5H),3.51-3.42(m,28H),3.37(d,J=7 .4Hz,3H),3.33(s,2H),2.88(s,2H),2.79(d,J=9.3Hz,8H),2.59(s,4H),2.54(s,3H),2.36(d,J=13.2Hz,1H),2. 15(t,J=7.0Hz,3H),1.99(dd,J=14.6,8.8Hz,4H),1.87(s,3H),1.70(s,1H),1.45(q,J=10.7,9.0Hz,5H),1.39(d ,J=6.9Hz,4H),1.27(dd,J=7.1,4.4Hz,4H),1.18(d,J=9.5Hz,3H),0.82(d,J=6.8Hz,3H),0.77(d,J=6.7Hz,3H). LC-MS m / z(ESI):922.6[M / 2+1] + .

[0559] Example A-25: Synthesis of compound DL-25

[0560] Following the preparation method of Example A-22, the starting material 1.7A was replaced with 1.5A to obtain the title product compound DL-25 (9.5 mg, 19% yield). 1H NMR (400MHz, DMSO-d6) δ10.95(s,1H),9.71(s,1H),8.55(d,J=3.2Hz,1H),8.05(d,J=7.0Hz,1H),7.99(s,1H),7.6 7(d,J=7.8Hz,1H),7.58(d,J=2.2Hz,2H),7.56(s,1H),7.53(s,1H),7.46(d,J=8.0Hz,1H),7.34(d,J=7.8Hz,1H),7 .30-7.22(m,2H),7.13(s,1H),7.06(s,1H),6.94(s,2H),6.81(dd,J=7.8,2.3Hz,1H),5.07(dd,J=13.2,5.1Hz,1H) ,4.95(s,2H),4.88(p,J=7.0Hz,1H),4.42(d,J=17.4Hz,1H),4.36-4.24(m,2H),4.09(dd,J=7.8,5.5Hz,1H),4.00( s,1H),3.67(d,J=14.6Hz,1H),3.59(d,J=6.7Hz,2H),3.47(d,J=5.0Hz,36H),3.41-3.37(m,4H),3.20(s,3H),2.95 -2.83(m,2H),2.79(d,J=9.1Hz,5H),2.55(dd,J=17.7,11.3Hz,3H),2.36(dt,J=13.1,3.7Hz,1H),2.15(t,J=7.4Hz ,2H),1.99(td,J=16.2,14.7,5.2Hz,3H),1.89(d,J=14.0Hz,2H),1.69(d,J=11.2Hz,1H),1.52-1.41(m,4H),1.39( d, J=7.0Hz, 3H), 1.28 (d, J=7.0Hz, 3H), 1.17 (dt, J=15.0, 8.2Hz, 3H), 0.83 (d, J=6.8Hz, 3H), 0.77 (d, J=6.8Hz, 3H). LC-MS m / z(ESI):810.5[M / 2+1] + .

[0561] Example A-26: Synthesis of compound DL-26

[0562] Following the preparation method of Example A-22, raw material 1.7A was replaced with 1.26B to obtain the title product DL-26 (21 mg, 7% yield). 1H NMR (400MHz, DMSO-d6) δ11.00 (s, 1H), 9.81 (s, 1H), 8.57 (d, J = 3.1Hz, 1H), 8.13 (d,J=7.1Hz,1H),8.03(s,1H),7.70(d,J=7.8Hz,1H),7.64-7.52(m,4H),7.48(d ,J=7.9Hz,1H),7.32(d,J=29.4Hz,3H),7.25-7.04(m,2H),6.98(s,1H),6.84(d, J=7.5Hz,1H),5.44(s,2H),5.11(dd,J=13.4,5.0Hz,1H),4.97(s,2H),4.93-4.7 4(m,2H),4.65-4.50(m,3H),4.44(d,J=16.3Hz,3H),4.30(d,J=17.4Hz,2H),4. 21-3.86(m,4H),3.82-3.39(m,40H),3.00-2.72(m,7H),2.68-2.52(m,4H),2.38 (q,J=14.8,14.3Hz,1H),2.16(d,J=7.7Hz,2H),2.11-1.78(m,3H),1.70(s,1H), 1.43 (dd, J=17.3, 10.3Hz, 7H), 1.35-1.11 (m, 7H), 0.81 (dd, J=24.6, 6.7Hz, 6H). LC-MS m / z (ESI): 856.5 [M / 2+1] + .

[0563] Example A-27: Synthesis of compound DL-27

[0564] Following the preparation method of Example A-13, compound 7 was replaced with compound 27 to obtain the title product compound DL-27 (22 mg, 9.42% yield). 1H NMR (400MHz, DMSO-d6) δ11.16(s,1H),10.97(s,1H),9.97(s,1H),8.54(d,J= 3.2Hz, 1H), 8.20 (dd, J=24.0, 7.3Hz, 2H), 7.80 (d, J=8.7Hz, 1H), 7.69 (d, J=7. 9Hz,1H),7.66-7.51(m,5H),7.48(d,J=7.9Hz,1H),7.28(d,J=8.0Hz,2H),7.1 6(s,1H),7.08(d,J=8.0Hz,1H),6.98(s,2H),6.80(d,J=7.5Hz,1H),5.10(dd, J=13.2,5.1Hz,1H),5.02-4.81(m,3H),4.64(s,1H),4.52-4.02(m,4H),3.55- 3.35(m,35H),3.23(s,3H),3.04(d,J=6.2Hz,2H),2.98-2.74(m,7H),2.62(t, J=18.9Hz,1H),2.46-2.28(m,1H),2.11(t,J=7.4Hz,2H),2.05-1.90(m,2H),1 .46(dt,J=24.8,7.5Hz,8H),1.34-1.11(m,7H),0.83(dd,J=18.7,6.8Hz,7H). LC-MS m / z (ESI): 837.4 [M / 2+23] + .

[0565] Example A-28: Synthesis of compound DL-28

[0566] Step 1: Intermediate 1.4H (116.29 mg, 132.46 μmol) and compound 31 (70 mg, 110.39 μmol) were dissolved in DMF (4 mL), and DIPEA (142.67 mg, 1.10 mmol) was added. The reaction mixture was stirred at room temperature for 3 hours. After evaporation to dryness, a brown oil was obtained. Silica gel column chromatography (DCM:MeOH = 12:1) yielded the brown oil compound DL-28-1 (130 mg, 85.78% yield). LC-MS m / z (ESI): 1272.6 [M-100+1] +

[0567] Step 2: Dissolve intermediate DL-28-1 (130 mg, 94.69 μmol) in DCM (2.5 mL). Slowly add TFA (431.85 mg, 3.79 mmol) dissolved in DCM (0.5 mL) to the reaction solution. Stir at room temperature for 1 hour. After concentration, obtain crude DL-28-2 (100 mg, crude product) as a brown oil. LC-MS m / z (ESI): 1272.6 [M+1] +

[0568] Step 3: Compound DL-28-2 (50 mg, 39.28 μmol) and intermediate 1.5A (19.73 mg, 43.21 μmol) were dissolved in DMF (3 mL), and DIPEA (25.38 mg, 196.41 μmol) and HATU (17.78 mg, 47.14 μmol) were added. The reaction mixture was stirred at room temperature for 3 hours. After concentration, the reaction solution was separated by acidic preparation (acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 20%-55%) to obtain compound DL-28 (26.5 mg, 39.18% yield); LC-MS m / z (ESI): 856.0 [M / 2+1] + ; 1H NMR (400MHz, DMSO-d6) δ10.99(s,1H),9.77(s,1H),8.57(s,1H),8.10(d,J=6.7Hz, 1H),8.01(s,1H),7.67(d,J=7.9Hz,1H),7.64-7.55(m,3H),7.53(s,1H),7.46(d,J =7.9Hz,1H),7.41-7.22(m,3H),7.19-7.09(m,1H),7.05(s,1H),6.96(s,1H),6.83 (d,J=7.4Hz,1H),5.09(dd,J=13.3,5.1Hz,1H),4.99(s,2H),4.88(t,J=7.1Hz,1H) ,4.42(d,J=17.3Hz,1H),4.35-4.23(m,2H),4.06(d,J=34.1Hz,2H),3.47(d,J=4.8 Hz,30H),3.41-3.34(m,26H),3.21(s,3H),3.02-2.71(m,6H),2.64-2.51(m,2H),2 .36(d,J=13.2Hz,1H),2.14(d,J=7.7Hz,2H),2.09-1.65(m,2H),1.42(dd,J=27.8, 7.1Hz,7H),1.29(d,J=7.1Hz,3H),1.23-1.08(m,1H),0.80(dd,J=23.5,6.7Hz,6H).

[0569] Example A-29: Synthesis of compound DL-29

[0570] Step 1: Compound 42 (315 mg, 461.82 μmol, TFA), intermediate 1.4H (405.45 mg, 461.82 μmol), DMF (5 mL), and N,N-diisopropylethylamine (298.43 mg, 2.31 mmol, 402.20 μL) were added to a reaction flask, and the reaction was carried out at 20 °C for 16 hours. The solvent was evaporated, and silica gel was added. The mixture was purified by column chromatography (DCM / MeOH, O to 20%) to give a yellow liquid DL-29-1 (210 mg, 34.79% yield). LC-MS m / z (ESI): 1206.5 [M+1-100] + .

[0571] Step 2: Add DL-29-1 (100 mg, 76.52 μmol), DCM (1 mL), and TFA (174.49 mg, 1.53 mmol) to the reaction flask and react at 20 °C for 2 hours. Remove the solvent by rotary evaporation and add directly to the next step to obtain the yellow liquid DL-29-2 (90 mg, 74.58 μmol, 97.47% yield). LC-MS m / z (ESI): 603.8 [M / 2+1] + .

[0572] Step 3: Add DL-29-2 (90 mg, 74.58 μmol), intermediate 1.5A (34.05 mg, 74.58 μmol), DMF (2 mL), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (28.14 mg, 74.58 μmol), and N,N-diisopropylethylamine (48.19 mg, 372.90 μmol, 64.95 μL) to the reaction flask and react at 20 °C for 2 hours. The solvent was evaporated and concentrated to obtain a yellow oil. The yellow oil was directly sent for preparative high-performance liquid chromatography (HPLC) for separation (acid method, Gilson, Waters-SunFire C18; ACN / Water + 0.04% FA; gradient, 15%-45%). After freeze-drying, a pale yellow solid compound DL-29 (6 mg, 4.89% yield) was obtained. LC-MS m / z (ESI): 823.0 [M / 2+1] + . 1H NMR(400MHz,DMSO-d6)δ10.99(s,1H),9.77(s,1H),8.81(s,1H),8.40(s,1 H),8.25(d,J=8.9Hz,1H),8.14-8.04(m,2H),7.70-7.51(m,5H),7.49-7.3 5(m,3H),7.26(d,J=8.2Hz,2H),7.12(dd,J=28.5,9.1Hz,3H),6.96(s,1H) ,5.09(dd,J=13.3,5.1Hz,1H),5.02-4.82(m,3H),4.48-4.24(m,3H),4.10 -3.97(m,3H),3.79(s,1H),3.66 -3.55(m,5H),3.47(s,38H),3.40(dd,J=5.9,3.6Hz,1H),3.21(s,4H),2.90-2.81(m,1H),2. 81-2.71(m,2H),2.40-2.27(m,1H),2.15(t,J=7.4Hz,2H),2.00(dd,J=13.6,7.9Hz,4H),1.75 -1.65(m,6H),1.50-1.35(m,6H),1.28(d,J=7.1Hz,3H),1.24-1.11(m,3H),0.80(dd,J=23.3,6.7Hz,6H).

[0573] Example A-30: Synthesis of compound DL-30

[0574] Step 1: Intermediate 1.4H (200 mg, 0.22 mmol) and compound 1 (124 mg, 0.22 mmol) were dissolved in DMF (5 mL), followed by the addition of DIPEA (57.2 mg, 0.44 mmol) and HATU (83.6 mg, 0.22 mmol). The reaction was carried out at room temperature for 2.0 h. After the reaction was completed, the product was directly concentrated by rotary evaporation to obtain a yellow oil. Separation was performed by silica gel column chromatography (dichloromethane:methanol = 100:1 to 10:1) to directly give a brown solid DL-30-1 (169 mg, 60% yield). LC-MS m / z (ESI): 1284.5 [M+1] + .

[0575] Step 2: Dissolve DL-30-1 (169 mg, 0.14 mmol) in DCM (5 mL), then add TFA (1 mL) and react at room temperature for 2.0 h. After the reaction, directly evaporate to dryness and concentrate to obtain a yellow oil DL-30-2 (165 mg, 100% yield), which can be directly used for the next step. LC-MS m / z (ESI): 1184.5 [M+1] + .

[0576] Step 3: Dissolve DL-30-2 (165 mg, 0.14 mmol) and intermediate 1.7A (88 mg, 0.14 mmol) in DMF (5 mL), then add DIPEA (36.4 mg, 0.28 mmol) and HATU (53.2 mg, 0.14 mmol), and react at room temperature for 1.5 h. After the reaction, directly evaporate to dryness and concentrate to obtain a yellow oil. The yellow oil was directly sent for preparative high performance liquid chromatography (Acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 24%-44%), and after freeze-drying, a pale yellow solid compound DL-30 (13 mg, 23.5% yield) was obtained. LC-MS m / z (ESI): 900.0 [M / 2+1] + . 1H NMR (400MHz, DMSO-d6) δ10.97(s,1H),9.71(s,1H),8.76(s,1H),8.05(d,J=6.9Hz,1 H),7.97(s,1H),7.62(d,J=1.8Hz,1H),7.53(dt,J=14.2,7.5Hz,4H),7.34(s,1H),7. 26(d,J=7.2Hz,2H),7.13(d,J=14.6Hz,2H),6.95(d,J=3.7Hz,2H),6.77(s,1H),5.09 (dd,J=13.2,5.1Hz,1H),4.96(s,2H),4.54(d,J=17.3Hz,1H),4.43-4.39(m,2H),4.3 5-4.30(m,1H),4.13-4.07(m,1H),4.00(s,1H),3.65(s,1H),3.59(d,J=6.5Hz,2H),3 .48(s,53H),3.40(d,J=5.2Hz,2H),3.34(d,J=6.9Hz,6H),3.21(s,3H),2.94-2.84(m ,2H),2.79(s,4H),2.64-2.52(m,3H),2.15(t,J=7.3Hz,2H),2.03-1.94(m,3H),1.45 (s,4H),1.28(d,J=6.9Hz,3H),1.19(d,J=16.2Hz,4H),0.80(dd,J=22.4,6.7Hz,6H).

[0577] Example A-31: Synthesis of compound DL-31

[0578] Intermediate DL-30-2 (55 mg, 0.046 mmol) and intermediate 1.5A (21.2 mg, 0.046 mmol) were dissolved in DMF (5 mL), followed by the addition of DIPEA (12 mg, 0.092 mmol) and HATU (17.5 mg, 0.046 mmol). The reaction was carried out at room temperature for 1.5 h. After the reaction was completed, the solution was directly concentrated by rotary evaporation to obtain a yellow oil. The yellow oil was directly subjected to preparative high-performance liquid chromatography (HPLC) for separation (acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 24%-44%). After lyophilization, a pale yellow solid compound DL-31 (12 mg, 16% yield) was obtained. LC-MS m / z (ESI): 811.9 [M / 2+1] + . 1H NMR (400MHz, DMSO-d6) δ10.98(s,1H),9.72(s,1H),8.77(s,1H),8.06(d,J=7.2Hz,1H),7. 98(s,1H),7.62(d,J=2.0Hz,1H),7.59-7.54(m,3H),7.52-7.48(m,1H),7.34(s,1H),7.27 (s,2H),7.13(d,J=17.6Hz,2H),6.95(s,2H),6.77(d,J=5.9Hz,1H),5.09(dd,J=13.4,5.1 Hz,1H),4.95(s,2H),4.54(d,J=17.3Hz,1H),4.44-4.36(m,3H),4.31(s,1H),4.11-4.07( m,1H),4.00(s,1H),3.60(dd,J=20.7,13.5Hz,4H),3.47(d,J=5.0Hz,31H),3.39(dd,J=5. 8,3.5Hz,3H),3.21(s,3H),2.94-2.85(m,2H),2.80(d,J=9.0Hz,5H),2.66-2.52(m,3H),2 .41(d,J=13.4Hz,1H),2.14(d,J=7.1Hz,2H),2.03-1.93(m,4H),1.70(s,3H),1.50-1.36( m,5H),1.28(d,J=7.1Hz,3H),1.18(dd,J=15.7,8.4Hz,4H),0.80(dd,J=22.7,6.8Hz,8H).

[0579] Example A-32: Synthesis of compound DL-32

[0580] Intermediate DL-30-2 (55 mg, 0.046 mmol) and intermediate 1.24B (30.4 mg, 0.046 mmol) were dissolved in DMF (5 mL), followed by the addition of DIPEA (12 mg, 0.092 mmol) and HATU (17.5 mg, 0.046 mmol). The reaction was carried out at room temperature for 1.5 h. After the reaction was completed, the solution was directly concentrated by rotary evaporation to obtain a yellow oil. The yellow oil was directly subjected to preparative high-performance liquid chromatography (HPLC) (acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 24%-44%). After freeze-drying, a pale yellow solid compound DL-32 (11 mg, 12.8% yield) was obtained. LC-MS m / z (ESI): 924.4 [M / 2+1] + . 1H NMR (400MHz, DMSO-d6) δ10.98(s,1H),9.73(s,1H),8.77(s,1H),8.04(d,J=51.7Hz,3H),7.63-7.47(m,8H),7.38(s,3H),7 .11(s,4H),6.95(s,1H),6.78(s,1H),5.30(s,1H),5.10(s,1H),4.95(s,2H),4.54(d,J=17.4Hz,2H),4.49-4.46(m,1H),4 .40(d,J=7.6Hz,4H),4.34(s,4H),4.10(s,2H),3.58(s,7H),3.47(d,J=5.6Hz,23H),2.79(s,12H),2.60(d,J=37.2Hz,7H) ,2.31-2.28(m,1H),2.15(s,2H),1.97(d,J=7.5Hz,6H),1.44(s,8H),1.28(d,J=6.9Hz,6H),0.79(dd,J=21.8,6.0Hz,12H).

[0581] Example A-33: Synthesis of compound DL-33

[0582] Step 1: Compound 33 (46 mg, 0.083 mmol) and intermediate 1.4H (72.7 mg, 0.083 mmol) were dissolved in DMF (5 mL), followed by the addition of DIPEA (20.8 mg, 0.16 mmol) and HATU (31.5 mg, 0.083 mmol). The reaction was carried out at room temperature for 2.0 h. After the reaction was completed, the product was directly concentrated by rotary evaporation to obtain a yellow oil. Separation was performed by silica gel column chromatography (dichloromethane:methanol = 100:1 to 10:1) to directly give a brown solid DL-33-1 (32 mg, 29.8% yield). MS m / z (ESI): 1294.58 [M+1] + .

[0583] Step 2: Dissolve DL-33-1 (32 mg, 0.025 mmol) in DCM (5 mL), then add TFA (1 mL) and react at room temperature for 2.0 h. After the reaction is complete, directly evaporate to dryness and concentrate to obtain a yellow oil DL-33-2 (25 mg, 83% yield), which can be directly used for the next step. MS m / z (ESI): 1194.5 [M+1] + .

[0584] Step 3: Dissolve DL-33-2 (25 mg, 0.021 mmol) and intermediate 1.5A (9.6 mg, 0.021 mmol) in DMF (5 mL), then add DIPEA (5.5 mg, 0.042 mmol) and HATU (8 mg, 0.021 mmol), and react at room temperature for 1.5 h. After the reaction, directly evaporate to dryness and concentrate to obtain a yellow oil. The yellow oil was directly sent for preparative high performance liquid chromatography (Acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 24%-44%), and after freeze-drying, a pale yellow solid compound DL-33 (8.5 mg, 23.5% yield) was obtained. MS m / z (ESI): 816.9 [M / 2+1] + . 1 H NMR (400MHz, DMSO-d6) δ10.98(s,1H),9.76(s,1H),8.56(s,1H),8.10(d,J=6.6Hz,1H),8.01(s,1H) ,7.68(d,J=8.1Hz,1H),7.58(d,J=9.8Hz,3H),7.53(s,1H),7.46(d,J=8.0Hz,1H),7.36(s,1H),7.2 6(d,J=7.8Hz,2H),7.16-7.04(m,2H),6.96(s,2H),6.83(d,J=7.1Hz,1H),5.09(dd,J=13.1,4.9Hz, 1H),4.96(s,2H),4.88(t,J=7.0Hz,1H),4.43(d,J=17.0Hz,1H),4.30(t,J=13.4Hz,2H),4.13-4.08( m,1H),4.02(s,1H),3.66(s,1H),3.58(t,J=6.4Hz,2H),3.47(d,J=5.1Hz,31H),3.41-3.38(m,2H), 3.29(s,4H),3.21(s,3H),3.19-3.15(m,2H),2.88(dd,J=21.4,9.5Hz,2H),2.78(s,2H),2.57(d,J=1 8.8Hz,3H),2.39-2.31(m,1H),2.14(d,J=7.2Hz,2H),2.03-1.94(m,4H),1.69(s,2H),1.50-1.35(m ,8H),1.29(d,J=7.1Hz,3H),1.22-1.12(m,5H),0.96(t,J=6.8Hz,3H),0.80(dd,J=23.3,6.7Hz,6H).

[0585] Example A-34: Synthesis of compound DL-34

[0586] Step 1: Compound 32 (51 mg, 0.089 mmol) and intermediate 1.4H (96.14 mg, 0.11 mmol) were dissolved in DMF (10 mL), followed by the addition of DIPEA (14.3 mg, 0.11 mmol) and HATU (41.8 mg, 0.11 mmol). The reaction was carried out at room temperature for 12.0 h. After the reaction was completed, the product was directly concentrated by rotary evaporation to obtain a yellow oil. Separation was performed by silica gel column chromatography (dichloromethane:methanol = 100:1 to 10:1) to directly give a brown solid DL-34-1 (25 mg, 21.5% yield). LC-MS m / z (ESI): 1308.6 [M+1] + .

[0587] Step 2: Dissolve DL-34-1 (25 mg, 0.019 mmol) in DCM (5 mL), then add TFA (1 mL) and react at room temperature for 2.0 h. After the reaction, directly evaporate to dryness and concentrate to obtain a yellow oil DL-34-2 (21 mg, 91.3% yield), which can be directly used for the next step. LC-MS m / z (ESI): 1208.6 [M+1] + .

[0588] Step 3: Dissolve DL-34-2 (21 mg, 0.017 mmol) and intermediate 1.5A (9.1 mg, 0.02 mmol) in DMF (10 mL), then add DIPEA (2.6 mg, 0.02 mmol) and HATU (7.6 mg, 0.02 mmol), and react at room temperature for 1.5 h. After the reaction, directly evaporate to dryness and concentrate to obtain a yellow oil. The yellow oil was directly sent for preparative high performance liquid chromatography (acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 24%-44%), and after freeze-drying, a pale yellow solid compound DL-34 (4.2 mg, 12.7% yield) was obtained. LC-MS m / z (ESI): 823.9 [M / 2+1] + . 1H NMR (400MHz, DMSO-d6) δ10.98(s,1H),9.77(s,1H),8.66(s,1H),8.06(d,J=25.4 Hz,2H),7.67(d,J=7.9Hz,1H),7.62-7.53(m,6H),7.46(d,J=8.1Hz,1H),7.37(s, 1H),7.26(d,J=7.5Hz,2H),7.07(s,2H),6.96(s,3H),5.09(dd,J=13.2,5.0Hz,1 H),4.96(s,2H),4.91-4.84(m,1H),4.43(d,J=16.7Hz,1H),4.32(d,J=10.0Hz,2H ),4.01(s,2H),3.58(s,2H),3.47(s,34H),3.40(d,J=5.3Hz,4H),3.21(s,5H),2 .93-2.71(m,6H),2.66-2.52(m,4H),2.33(d,J=19.5Hz,2H),2.15(s,2H),2.01-1 .95(m,4H),1.45(s,4H),1.39(d,J=6.9Hz,3H),1.29(d,J=7.1Hz,4H),1.16(d,J =6.0Hz,2H),1.03(d,J=6.6Hz,5H),0.82(d,J=5.8Hz,3H),0.77(d,J=6.7Hz,2H).

[0589] Example A-35: Synthesis of compound DL-35

[0590] Step 1: Intermediate 1.4H (150 mg, 0.17 mmol) and compound 28 (140 mg, 0.25 mmol) were dissolved in DMF (10 mL), followed by the addition of DIPEA (44.2 mg, 0.34 mmol) and HATU (64.6 mg, 0.17 mmol). The reaction was carried out at room temperature for 12.0 h. After the reaction was completed, the product was directly concentrated by rotary evaporation to obtain a yellow oil. Separation was performed by silica gel column chromatography (dichloromethane:methanol = 100:1 to 10:1) to directly give a brown solid DL-35-1 (134.6 mg, 61.0% yield). LC-MS m / z (ESI): 1198.5 [M+1-100] + .

[0591] Step 2: Dissolve DL-35-1 (50 mg, 0.04 mmol) in DCM (5 mL), then add TFA (1 mL) and react at room temperature for 2.0 h. After the reaction is complete, directly evaporate to dryness and concentrate to obtain a yellow oily substance DL-35-2 (38 mg, 79% yield), which can be directly used for the next step. LC-MS m / z (ESI): 1198.5 [M+1] + .

[0592] Step 3: Dissolve DL-35-2 (38 mg, 0.032 mmol) and intermediate 1.5A (14.5 mg, 0.032 mmol) in DMF (10 mL), then add DIPEA (9.4 mg, 0.072 mmol) and HATU (13.7 mg, 0.036 mmol), and react at room temperature for 2.0 h. After the reaction, directly evaporate to dryness and concentrate to obtain a yellow oil. The yellow oil was directly sent for preparative high performance liquid chromatography (acid method, GILSON, Waters-SunFire C18 column; ACN / Water + 0.04% FA; gradient, 24%-44%), and after freeze-drying, a pale yellow solid compound DL-35 (12.0 mg, 32.0% yield) was obtained. LC-MS m / z (ESI): 818.9 [M / 2+1] + . 1 H NMR(400MHz,DMSO-d6)δ10.96(s,1H),9.71(s,1H),8.75(s,1H),8.05(d,J =6.2Hz,1H),7.96(s,1H),7.61(s,1H),7.59-7.53(m,3H),7.50(d,J=6.0Hz ,1H),7.32(s,1H),7.24(d,J=7.7Hz,2H),7.12(d,J=14.5Hz,2H),6.93(s,2 H),6.77(s,1H),5.07(s,1H),4.94(s,2H),4.53(d,J=17.4Hz,1H),4.42-4. 29(m,4H),4.08(s,2H),3.69-3.56(m,3H),3.46(s,36H),3.39(d,J=4.4Hz ,5H),3.18(d,J=12.6Hz,5H),2.92-2.75(m,4H),2.58(s,2H),2.39(s,1H), 2.14(s,2H),1.98(s,3H),1.88(d,J=12.5Hz,3H),1.43(s,5H),1.27(d,J=6 .9Hz,3H),1.19(d,J=8.8Hz,3H),0.96(t,J=6.6Hz,3H),0.84-0.75(m,5H).

[0593] Example A-36: Synthesis of compound DL-36

[0594] Step 1: Intermediate 1.4H (149 mg, 144.26 μmol) and compound 35 (111.69 mg, 145.66 μmol) were dissolved in DMF (2.0 mL), and DIPEA (55.93 mg, 432.78 μmol, 75.38 μL) was added. The mixture was stirred at 40 °C for 16 hours. The reaction solution was then dried under reduced pressure using an oil pump. The crude product was purified by Combi Flash silica gel column chromatography (4 g, 0–9.0% MeOH / DCM) to obtain a colorless, viscous solid DL-36-1 (175 mg, 78.94% yield). LC-MS m / z (ESI): 1336.5 [M+1] + .

[0595] Step 2: Dissolve DL-36-1 (85 mg, 55.31 μmol) in DCM (1.2 mL), stir at room temperature, and slowly add a solution of TFA (378.41 mg, 3.32 mmol) dissolved in DCM (0.3 mL). After the addition is complete, continue stirring for 30 min. Dilute the reaction solution with DCM (5.0 mL) and evaporate to dryness under reduced pressure. Then evaporate to dryness under reduced pressure using an oil pump to obtain a colorless, viscous solid DL-36-2 (85 mg, 99.40% yield). LC-MS m / z (ESI): 1236.5 [M+1] + .

[0596] Step 3: DL-36-2 (80 mg, 54.98 μmol) and intermediate 1.5A (28 mg, 61.33 μmol) were dissolved in DMF (1.5 mL), and DIPEA (21.32 mg, 164.94 μmol, 28.73 μL) was added. The mixture was stirred at 20 °C, and then HATU (25.09 mg, 65.98 μmol) was added. After the addition was complete, the mixture was stirred for another 2 hours. The reaction solution was dried under reduced pressure using an oil pump at 40 °C. The crude product was purified by HPLC (acid method, GILSON, Waters-Sun Fire C18 column; ACN / Water-0.04% FA; 5%-96%), and after lyophilization, a white solid compound DL-36 (7.14 mg, 7.75% yield, 100.00% purity) was obtained. LC-MS m / z (ESI): 838.0 [M / 2+1] + . 1H NMR (400MHz, DMSO-d6) δ11.00(s,1H),9.78(s,1H),8.65(s,1H),8.16-8.09(m,1H),8.03(s,1H),7. 67(d,J=9.2Hz,1H),7.62(d,J=18.0Hz,2H),7.58(s,1H),7.53(s,1H),7.46(d,J=8.4Hz,1H),7.37( d,J=8.4Hz,1H),7.27(d,J=8.3Hz,2H),7.12(s,1H),7.06(d,J=8.4Hz,1H),6.96(s,2H),6.94-6.89 (m,1H),5.09(dd,J=5.6Hz,8.4Hz,1H),4.97(s,2H),4.86(d,J=7.6Hz,1H),4.47-4.37(m,3H),4.34- 4.25(m,3H),4.10(t,J=6.7Hz,2H),4.04-3.98(m,2H),3.62-3.57(m,7H),3.56-3.44(m,38H),3.20 (s,3H),2.92-2.86(m,1H),2.77(s,2H),2.66-2.57(m,2H),2.39-2.31(m,2H),2.15(t,J=7.5Hz,2H ),2.06-1.94(m,4H),1.88-1.78(m,2H),1.74-1.65(m,1H),1.53-1.42(m,4H),1.39(t,J=6.7Hz,3H ), 1.28 (d, J = 7.3Hz, 3H), 1.18 (dd, J = 16.6, 9.2Hz, 3H), 0.82 (d, J = 6.8Hz, 3H), 0.77 (d, J = 6.8Hz, 3H).

[0597] Example A-38: Synthesis of compound DL-38

[0598] Intermediate 1.9C (93 mg, 37.15 μmol), intermediate 1.24B (26 mg, 38.14 μmol), and DIPEA (48.01 mg, 371.47 μmol, 64.70 μL) were dissolved in DMF (2.0 mL), stirred at 20 °C, and HATU (16.95 mg, 44.58 μmol) was added. The reaction mixture was stirred for another 30 min. The reaction solution was evaporated to dryness under reduced pressure at room temperature. The crude product was purified by HPLC (acid method, GILSON, Waters-Sun Fire C18 column; ACN / Water-0.04% FA; 5%-95%), and lyophilized to give a white solid compound DL-38 (20.90 mg, 28.14% yield, 97.07% purity). LC-MS m / z (ESI): 970.5 [M / 2+1] + . 1H NMR (400MHz, DMSO-d6) δ10.97(s,1H),9.73(s,1H),8.80(s,1H),8.17(s,1H),8.07(d,J=6.9Hz,1H),7.9 9(s,1H),7.62(s,1H),7.58(d,J=9.0Hz,2H),7.54(s,1H),7.53-7.47(m,1H),7.34(d,J=7.6Hz,1H),7.27 (t,J=10.4Hz,2H),7.13(d,J=17.6Hz,2H),6.94(s,2H),6.81(d,J=5.7Hz,1H),5.08(dd,J=13.3,5.1Hz, 1H),4.99(s,2H),4.56(s,1H),4.52(s,1H),4.45(s,1H),4.44-4.37(m,4H),4.36-4.25(m,4H),4.10(t,J =6.7Hz,2H),4.00(s,2H),3.70-3.61(m,5H),3.60-3.53(m,9H),3.47(d,J=4.9Hz,31H),2.92(d,J=25.8 Hz,6H),2.83-2.75(m,3H),2.69(dd,J=14.7,6.8Hz,3H),2.62-2.56(m,3H),2.53(t,J=7.1Hz,4H),2.44- 2.34(m,2H),2.15(t,J=7.3Hz,2H),2.08-1.93(m,4H),1.89(d,J=16.6Hz,2H),1.70(s,1H),1.46(dq,J= 14.4,7.7Hz,4H),1.28(d,J=7.0Hz,3H),1.24-1.11(m,3H),0.82(d,J=6.7Hz,3H),0.77(d,J=6.7Hz,3H).

[0599] Example A-39: Synthesis of compound DL-39

[0600] Intermediate DL-39-1 (65 mg, 48.35 μmol) and intermediate 1.5A (24.62 mg, 53.94 μmol) were dissolved in DMF (1.5 mL), and DIPEA (31.24 mg, 241.75 μmol, 42.11 μL) was added. The mixture was stirred at 20 °C, and then HATU (22.06 mg, 58.02 μmol) was added. After the addition was complete, the mixture was stirred for another hour. The reaction mixture was then dried under reduced pressure using an oil pump at 40 °C. The crude product was purified by HPLC (acid method, GILSON, Waters-Sun Fire C18 column; ACN / Water-0.04% FA; 7%-95%), and lyophilized to give a white solid compound DL-39 (12.88 mg, 15.79% yield, 99.30% purity). LC-MS m / z (ESI): 838.0 [M / 2+1] + . 1H NMR (400MHz, DMSO-d6) δ10.95(s,1H),9.73(s,1H),8.51(d,J=3.3Hz,1H),8.06(d,J=7.0Hz,1H),7.98(s,1H),7.66(d,J=7 .9Hz,1H),7.58(d,J=2.1Hz,2H),7.54(d,J=16.1Hz,2H),7.45(d,J=7.9Hz,1H),7.33(d,J=7.8Hz,1H),7.26(d,J=8.2Hz,2H ),7.12(d,J=8.4Hz,1H),7.04(d,J=8.2Hz,1H),6.94(d,J=1.4Hz,2H),6.78(d,J=7.4Hz,1H),5.07(dd,J=13.1,5.1Hz,1H) ,4.96(s,2H),4.87(t,J=7.1Hz,1H),4.42(d,J=17.4Hz,2H),4.28(d,J=17.8Hz,2H),4.09(t,J=6.7Hz,1H),4.00(s,1H),3. 77(s,1H),3.66(d,J=14.4Hz,1H),3.58(d,J=6.6Hz,4H),3.46(d,J=4.8Hz,31H),3.43-3.36(m,4H),3.36-3.33(m,2H),3. 27-3.22(m,3H),3.20(d,J=1.5Hz,3H),3.18-3.09(m,1H),2.94-2.82(m,2H),2.82-2.73(m,2H),2.61-2.50(m,3H),2.35(d ,J=13.3Hz,1H),2.14(t,J=7.4Hz,2H),1.99(dt,J=15.6,7.8Hz,5H),1.88-1.77(m,2H),1.75-1.61(m,1H),1.44(q,J=11.1 ,9.2Hz,4H),1.38(d,J=6.9Hz,3H),1.27(d,J=7.0Hz,3H),1.23-1.09(m,3H),0.82(d,J=6.7Hz,3H),0.76(d,J=6.8Hz,3H).

[0601] Example B-1: Synthesis of conjugates ADC01 to ADC19, ADC21 to ADC36, ADC38 and ADC39

[0602] Two equivalents of tri-(2-carboxyethyl)phosphine (TCEP) were added to a pertuzumab (2 mg / ml) PBS / 1.0 mM EDTA (pH 6.8) buffer solution to treat the antibody solution, and the solution was incubated at 25°C for 3 hours to reduce interchain disulfide bonds. Compounds DL-1 to DL-19 prepared in Examples A-1 to A-19, compounds DL-21 to DL-36 prepared in Examples A-21 to A-36, and compounds DL-38 to DL-39 prepared in Examples A-38 to A-39 were dissolved in DMSO. Ten equivalents of DMSO solution of compounds DL-1 to DL-19, DL-21 to DL-36, and DL-38 to DL-39 were slowly added to the reduced antibody solution, and the reaction was stirred at 25°C for 1-4 hours. After the reaction was complete, the sample was filtered through a 0.22 μm membrane, and uncoupled compounds were removed using a desalting column with a 25 mM-His buffer (pH 5.5). The aggregation degree of the ADC was determined by size exclusion chromatography, and the DAR value of the ADC was determined by LC-MS. The structures and aggregation degrees of the obtained conjugates are shown in Tables 1 and 2, respectively.

[0603] Example B-2: Synthesis of conjugate ADC20

[0604] Using the same method as in Example B-1, compound DL-20 was conjugated with pertuzumab to obtain ADC20, and its DAR value and aggregation degree were detected. The test results are shown in Tables 1 and 2. Compound DL-20 was prepared according to the method in patent WO2021198965A1, and its structure is shown below:

[0605] Table 1: Structure and characterization of couplings

[0606] Table 2: Aggregation degree of couplings

[0607] As can be seen from Table 2, the conjugates of this application have a lower degree of aggregation.

[0608] Biological testing

[0609] Test Example 1: Test of the inhibitory activity of the compound on the proliferation of MDA-MB-453 cells

[0610] Taking the compounds in the embodiments of this application and control compound a as examples, the inhibitory activity against the proliferation of human breast ductal carcinoma cells MDA-MB-453 was tested to evaluate the in vitro cell activity of the compounds. The structure of control compound a is shown below, and it can be prepared with reference to existing literature or obtained by commercial purchase.

[0611] I. Reagents required for the experiment

[0612] II. Boards and instruments required for the experiment

[0613] III. Experimental Methods

[0614] 1. MDA-MB-453 cells in the logarithmic growth phase were seeded into 384 cell culture plates except for the negative control wells to obtain compound test wells and positive control wells. The culture medium was 45 μL of IMDM medium containing 10 vol% FBS, and the cell density was 2000 / well. The negative control wells contained 45 μL of IMDM medium containing 10 vol% FBS without cells.

[0615] 2. Prepare a 10-fold solution of the desired compound to achieve the final concentration using 100% DMSO;

[0616] 3. Add 5 μL of the prepared test compound solution to the compound test well, and add 5 μL of 2.5 vol% DMSO solution to the negative control well and the positive control well;

[0617] 4. Place the cell culture plates in a 37°C, 5% CO2 cell culture incubator for 3 days;

[0618] 5. Remove from the incubator and let stand at room temperature for 0.5 hours to allow the cell culture plate to return to room temperature;

[0619] 6. Remove the CTG reagent from the refrigerator beforehand to thaw and allow it to return to room temperature;

[0620] 7. Add 40 μL of CTG reagent to each well of the cell culture plate;

[0621] 8. Incubate the cell culture plates at room temperature in the dark for 0.5 hours;

[0622] 9. Use a Tecan microplate reader to read the fluorescence signal value.

[0623] IV. Data Calculation

[0624] 1. PC wells are positive control wells, i.e., the wells with the maximum value measured on day 3, and NC wells are negative control wells, i.e., the wells with the minimum value measured on day 3.

[0625] 2. Calculation of compound inhibition rate

[0626] Where Inhibition ratio represents the inhibition rate of the tested compound, Signal Test This indicates the signal value of the cell after 72 hours of treatment with the detection compound. Mean of NC The value of the negative control well is represented by , and the SignalMean of PC is represented by the signal value of the positive control well. The data was fitted using XLFIT 5.0 software (IDBS, UK), with the logarithm of compound concentration as the X-axis and the inhibition rate as the Y-axis. A four-parameter model was used to calculate the half-maximal inhibitory concentration (IC50) of the compound. 50 .

[0627] The results of the inhibitory activity test of the compounds in the embodiments of this application on the proliferation of MDA-MB-453 cells are shown in Table 3:

[0628] Table 3

[0629] Test results show that, compared with control compound a, the compound of this application has significantly enhanced proliferative inhibitory activity against MDA-MB-453 cells.

[0630] Test Example 2: Degradation of GSPT1 protein in MDA-MB-453 cells by the compound

[0631] I. Reagents

[0632] II. Consumables and Instruments

[0633] III. Experimental Procedure

[0634] 1. Before testing, the 384-well culture plate was coated with 20 μg / mL PDL solution. After coating, the culture plate was sealed with film and stored at 4°C.

[0635] 2. MDA-MB-453 cells were passaged in complete medium (IMDM + 10% FBS) and maintained in the logarithmic growth phase. Before seeding, cells were washed with PBS, digested with TE, and counted. They were then seeded at a density of 2,000 cells / well into pre-coated 384-well culture plates (containing compound test wells, negative control wells, and positive control wells, where the negative control wells were cell-free, and the compound test wells and positive control wells contained cells). The plates were then incubated at 37°C and 5% CO2 for 4 hours.

[0636] 3. Prepare a gradient dilution buffer for the compound using DMSO. Add the gradient dilution buffer to the test wells of the compound, and add IMDM containing DMSO to the negative control wells and positive control wells. The final concentration of DMSO in all wells should be 0.25 vol%. Then, place the wells in an incubator and incubate at 37°C and 5% CO2 for 24 hours.

[0637] 4. Remove the culture plate and add fixative (PBS containing 2.5 vol% GA) directly to all wells. Fix the cells at room temperature for 30 minutes, then discard the culture medium.

[0638] 5. Add calibration staining solution (PBS containing 40 μg / mL Hoechst 33342 and 0.1 vol% Triton X-100) to all wells, stain at room temperature in the dark for 30 minutes, then discard the staining solution and immediately read the fluorescence signal value (excitation wavelength) on a microplate reader. 360nm / Emission wavelength 460nm );

[0639] 6. Add antigen retrieval solution (aqueous solution containing 1 mM EDTA) to all wells, place on a microplate shaker and heater, and perform high-temperature retrieval for 2 hours. Then remove the culture plate, allow it to cool to room temperature, and discard the retrieval solution.

[0640] 7. Prepare antibody dilution buffer (TBST containing 1 wt% BSA) and add it to the negative control wells. Then add GSPT1 antibody to the antibody dilution buffer at a ratio of 1:1000 and add it to the remaining wells except the negative control wells. After sealing with film, incubate at 4°C overnight, then discard all liquid.

[0641] 8. Wash all wells with TBST solution, repeating three times. Prepare antibody dilution buffer (TBST containing 1 wt% BSA), add anti-rabbit IgG-HRP conjugate antibody to the antibody dilution buffer at a ratio of 1:500, and add to all wells. Incubate at room temperature for 2 hours, then discard all liquid.

[0642] 9. Wash all wells with TBST solution, repeating three times. Then add TMB substrate to all wells and incubate at room temperature in the dark for 20 minutes to develop color. Immediately after incubation, add 2M sulfuric acid to stop the color development reaction. Read the signal on a microplate reader: absorbance value (OD). 450nm ).

[0643] IV. Data Calculation

[0644] 1. Before analysis, all data will be homogenized according to the signal in each well, as follows:

[0645] Where TMB values(OD) 450 nm This represents the absorbance value produced by the cells. Please refer to Experiment 9, Hoechst 33342 values(Fluorescence). 460 nm The value represents the fluorescence signal of the cell. Please refer to Experiment 5. Max(Hoechst 33342value) represents the fluorescence signal value of the positive control well.

[0646] 2. The calculation method for compound degradation rate is as follows:

[0647] Where DR% represents the degradation rate of the target protein by the tested compound, Dignal Test The SignalMean of NC indicates the signal generated by cells treated with the compound, and the SignalMean of PC indicates the signal generated by the positive control well.

[0648] 3. Fit the compound concentration and DR% using a four-parameter model, as follows:

[0649] DR% PC The percentage (DR%) indicates the degradation rate of the target protein in the positive control wells. NC The negative control well represents the degradation rate of the target protein, LogCmpd represents the logarithm of the compound concentration, and Hillslope represents the slope value obtained from curve fitting. The half-maximal concentration (DC) of different compounds is calculated using this model. 50 The values ​​were calculated and compared. The test results for representative compounds are shown in Table 4 below:

[0650] Table 4

[0651] As can be seen from the results in the table above, the compounds in this application have strong GSPT1 protein degradation activity.

[0652] Test Example 3: Pfeiffer Cell Selectivity Experiment

[0653] I. Reagents

[0654] II. Consumables and Instruments

[0655] III. Experimental Procedure

[0656] 1. Before testing, the 384-well culture plate was coated with 20 μg / mL PDL solution. After coating, the culture plate was sealed with film and stored at 4°C.

[0657] 2. Pfeiffer cells were passaged in complete medium (RPMI 1640 + 10% FBS) and maintained in the logarithmic growth phase. Before seeding, cells were harvested and counted, and seeded at a density of 10,000 cells / well into pre-coated 384-well culture plates (containing compound test wells, negative control wells, and positive control wells, where the negative control wells were cell-free, and the compound test wells and positive control wells contained cells). The plates were then incubated at 37°C and 5% CO2 for 4 hours.

[0658] 3. Prepare a gradient dilution buffer for the compound using DMSO. Add the gradient dilution buffer to the test wells of the compound, and add RPMI 1640 containing DMSO to the negative control wells and positive control wells. The final concentration of DMSO in all wells should be 0.1 vol%. Then place the wells in an incubator and incubate at 37°C and 5% CO2 for 24 hours.

[0659] 4. Remove the culture plate and add fixative (PBS containing 2.5% GA) directly to all wells. Fix the cells at room temperature for 30 minutes, then discard the culture medium.

[0660] 5. Add calibration staining solution (PBS containing 40 μg / mL Hoechst 33342 and 0.1% Triton X-100) to all wells, stain at room temperature in the dark for 30 minutes, then discard the staining solution and immediately read the fluorescence signal value (excitation wavelength) on a microplate reader. 360nm / Emission wavelength 460nm );

[0661] 6. Add antigen retrieval solution (containing 1× borate buffer) to all wells, place on a microplate shaker and heater, and perform high-temperature retrieval for 2 hours. Then remove the culture plate, let it cool to room temperature, and discard the retrieval solution.

[0662] 7. Prepare antibody dilution buffer (TBST containing 1% BSA) and add it to the negative control wells. Then, add GSPT1 / IKZF1 / IZKF3 / CK1α antibody to the antibody dilution buffer at a ratio of 1:1,000 and add it to the remaining wells except the negative control wells. After sealing with film, incubate at 4°C overnight, and then discard all liquid.

[0663] 8. Wash all wells with TBST solution, repeating three times. Prepare antibody dilution buffer (TBST containing 1 wt% BSA), add anti-rabbit IgG-HRP conjugate antibody to the antibody dilution buffer at a ratio of 1:500, and add to all wells. Incubate at room temperature for 2 hours, then discard all liquid.

[0664] 9. Wash all wells with TBST solution, repeating three times. Then add TMB substrate to all wells and incubate at room temperature in the dark for 20 minutes to develop color. Immediately after incubation, add 2M sulfuric acid to stop the color development reaction. Read the absorbance value (OD) on a microplate reader. 450 nm ).

[0665] IV. Data Calculation

[0666] 1. Before analysis, all data will be homogenized according to the signal in each well, as follows:

[0667] Where TMB values(OD) 450 nm This represents the absorbance value produced by the cells. Please refer to Experiment 9, Hoechst 33342 values(Fluorescence). 460 nm The value represents the fluorescence signal of the cell. Please refer to Experiment 5. Max(Hoechst 33342value) represents the fluorescence signal value of the positive control well.

[0668] 2. The calculation method for compound degradation rate is as follows:

[0669] Where DR% represents the degradation rate of the target protein by the test compound, Signal Test The SignalMean of NC indicates the signal generated by cells treated with the compound, and the SignalMean of PC indicates the signal generated by the positive control well.

[0670] 3. Fit the compound concentration and DR% using a four-parameter model, as follows:

[0671] DR% PC The percentage (DR%) indicates the degradation rate of the target protein in the positive control wells. NC The negative control well represents the degradation rate of the target protein, LogCmpd represents the logarithm of the compound concentration, and Hillslope represents the slope value obtained from curve fitting. The DC of different compounds is calculated using this model. 50The values ​​were calculated and compared. The test results are shown in Table 5.

[0672] Table 5: Inhibitory activity of compounds against IKZF1 and IKZF3

[0673] Test results show that, compared with control compounds a and b, the compounds of this application have better selectivity for IKZF1 and IKZF3. The structure of control compound b is shown below, and it can be prepared with reference to existing literature or obtained by commercial purchase.

[0674] Test Example 4: Pharmacokinetic Study in Mice

[0675] The drug concentration in the plasma of mice at different time points after intravenous (IV) administration of the compound of the present invention was determined by LC / MS / MS to study the pharmacokinetic behavior of the compound of the present invention in mice and evaluate its pharmacokinetic characteristics.

[0676] Reagents:

[0677] 1. Experimental Protocol: Healthy adult male ICR mice, weighing 25-40g, were used as experimental animals, 3 mice per group. All mice had free access to food and water. Animals were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. or Shanghai Jihui Laboratory Animal Breeding Co., Ltd. After arrival at the animal facility, animals were allowed a minimum acclimatization period of 3 days. The SPF (Spectrum Discharge) area was maintained at a temperature of 20-26℃ and a relative humidity of 40-70%. Illumination was provided by fluorescent lamps, with 12 hours of illumination (08:00-20:00) and 12 hours of no illumination, 2-5 mice per cage (same treatment group). Unlimited access to feed (irradiated and sterilized, purchased from Shaanxi Qinle Pharmaceutical Chemical Co., Ltd., China) and water (tap water purified by an ultrapure water filtration system) was provided. The specific dosing regimen is shown in the table below.

[0678] 2. Animal experimental prescription:

[0679] Solvent: 5% DMSO / 10% Solutol HS15 / 85% water (all by volume); Preparation of drug administration formulation: Intravenous group: Accurately weigh an appropriate amount of the test compound, add 5% of the formulation volume of DMSO, shake and sonicate to dissolve it completely, add 10% of the formulation volume of Solutol HS15 after dissolution, vortex for 2 minutes, and finally add 85% of the formulation volume of pure water, and sonicate to obtain a clear and transparent solution with a concentration of 0.4 mg / mL.

[0680] 3. Administration method: Select animals that meet the experimental requirements before administration, weigh and mark them. ICR mice are administered 2 mg / kg via tail vein.

[0681] 4. Sample Collection: Before sampling, mice were tethered. Blood was collected from each drug-treated mouse at predetermined time points: 0.083, 0.25, 0.5, 1, 2, 4, 7, and 24 hours after drug administration, for a total of 8 time points. Approximately 80 μL of blood was collected via the submandibular vein. The blood was transferred to 1.5 mL test tubes pre-filled with K2EDTA and centrifuged for 3 min (10000 rpm, 4℃) using an Eppendorf Centrifuge 5804R centrifuge to separate the plasma. The entire process was completed within 15 minutes of blood collection. All samples were stored at -20℃ (Panasonic) until sample analysis.

[0682] 5. Sample Analysis and Calculation: Drug concentration was determined using LC / MS / MS (SCIEX, TRIPLE QUAD 6500+), and pharmacokinetic parameters, such as the half-life (T0.05) of the drug, were calculated using Data Analysie System software (Shanghai Bojia Pharmaceutical Technology Co., Ltd., version 3.0). 1 / 2 Drug exposure AUC 0-t Drug clearance rate CLz, etc. The pharmacokinetic properties of some of the compounds in the embodiments of this invention in mice at the same dose and administration route are shown in Table 6:

[0683] Table 6. Pharmacokinetic parameters of the compounds in mice.

[0684] As can be seen from the data in the table above, compared with control compound a, the compound represented by this application has superior in vivo pharmacokinetic parameters.

[0685] Test Example 5: Assay on the inhibitory activity of the conjugate (ADC) on the proliferation of BT474 tumor cells

[0686] I. Reagents required for the experiment

[0687] II. Boards and instruments required for the experiment

[0688] III. Experimental Procedure

[0689] 1. BT-474 cells in the logarithmic growth phase were seeded into 384 cell culture plates except for the negative control wells to obtain compound test wells and positive control wells. The culture medium was 45 μL of RPMI 1640 medium containing 10 vol% FBS, and the cell density was 3000 / well. The negative control wells contained 45 μL of RPMI 1640 medium containing 10 vol% FBS without cells.

[0690] 2. Prepare a 10-fold solution of the desired compound to achieve the final concentration using 100% DMSO;

[0691] 3. Add 5 μL of the prepared test compound solution to the compound test well, and add 5 μL of 1 vol% DMSO solution to the negative control well and the positive control well;

[0692] 4. Place the cell culture plates in a 37°C, 5% CO2 cell culture incubator and culture for 5 days;

[0693] 5. Remove from the incubator and let stand at room temperature for 0.5 hours to allow the cell culture plate to return to room temperature;

[0694] 6. Remove the CTG reagent from the refrigerator beforehand to thaw and allow it to return to room temperature;

[0695] 7. Add 40 μL of CTG reagent to each well of the cell culture plate;

[0696] 8. Incubate the cell culture plates at room temperature in the dark for 0.5 hours;

[0697] 9. Use a Tecan microplate reader to read the Luminescence value.

[0698] IV. Data Calculation

[0699] 1. PC wells are positive control wells, i.e., the wells with the maximum value measured on day 5, and NC wells are negative control wells, i.e., the wells with the minimum value measured on day 5.

[0700] 2. Calculation of compound inhibition rate

[0701] Where Inhibition ratio represents the inhibition rate of the test compound, Test group represents the signal produced by cells treated with the compound, NC group represents the signal produced by the negative control well, and PC group represents the signal produced by the positive control well. Conjugates ADC01 to ADC19 and ADC20 were tested as one batch, conjugates ADC21 to ADC27 and ADC20 were tested as one batch, and conjugates ADC28 to ADC36 and ADC38 to ADC39 were tested as one batch. The test results of the three batches are shown in Tables 7, 8 and 9, respectively.

[0702] Table 7: Results of the inhibitory activity of ADC on the proliferation of BT474 tumor cells

[0703] Table 8: Results of ADC's inhibitory activity against BT474 tumor cell proliferation

[0704] Table 9: Results of the inhibitory activity of ADC on the proliferation of BT474 tumor cells

[0705] Test results show that the conjugate of this application has significantly enhanced proliferative inhibitory activity against BT474 cells.

[0706] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

[0707] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0708] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

A compound of formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: in, R1 is hydrogen or halogen; R2 and R3 are each independently hydrogen or C. 1-8 Alkyl (preferably C) 1-6 Alkyl, more preferably C 1-3 alkyl); Furthermore, R1, R2, and R3 are not all hydrogen at the same time; n is 1, 2, or 3; R 41 R 42 R 43 R 44 Each can be either hydrogen or halogen; R5, R6, R a R b R c R d R e R f m is selected from the following group of definitions: (i) R5 and R6 are each independently hydrogen and C. 1-8 Alkyl, C 3-6 cycloalkyl, -C 1-4 Alkylene-C 3-6 cycloalkyl, halogenated C 1-8 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-4 Alkylene-R7, the C 1-8 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-8 alkyl; R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-4 Alkyl, -S(O)-C 1-4 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens; R a R b R c R d R e R f Each independently is hydrogen, C 1-8 Alkyl or halogen; m is 0, 1, or 2; (ii) R6 is hydrogen, C 1-8 Alkyl, C 3-6 cycloalkyl, -C 1-4 Alkylene-C 3-6 cycloalkyl, halogenated C 1-8 Alkyl, or 4- to 6-membered heterocyclic alkyl, wherein the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-8 alkyl; R a R b R d R e R f Each independently is hydrogen, C 1-8 Alkyl or halogen; R c The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens; m is 0, 1, or 2; (iii) R6 is hydrogen, C 1-8 Alkyl, C 3-6 cycloalkyl, -C 1-4 Alkylene-C 3-6 cycloalkyl, halogenated C 1-8 Alkyl, or 4- to 6-membered heterocyclic alkyl, wherein the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-8 alkyl; R a R b R d R c R f Each independently is hydrogen, C 1-8 Alkyl or halogen; R e The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens; m can be 0, 1, or 2. The compound as claimed in claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R1 is a halogen; R2 and R3 are each hydrogen independently. The compound as claimed in claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R1 is fluorine; R2 and R3 are each hydrogen independently. The compound as claimed in claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R1 is hydrogen; R2 is hydrogen; R3 is C. 1-3 alkyl. The compound as claimed in claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R5 is hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl (preferably fluorinated C) 1-3 Alkyl), 4- to 6-membered heterocyclic alkyl or -C 1-2 Alkylene-R7, the C 1-3 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl; R6 is hydrogen; R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens; R a R b R c R d R e R f Each independently is hydrogen, C 1-3 Alkyl or halogen; m is 0. The compound as claimed in claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R6 is hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl, or 4- to 6-membered heterocyclic alkyl, wherein the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl; R a R b R d R e R f Each independently is hydrogen, C 1-3 Alkyl or halogen; R c The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens; m is 0 or 1. The compound as claimed in claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R6 is hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl, or 4- to 6-membered heterocyclic alkyl, wherein the C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl; R a R b R d R c R f Each independently is hydrogen, C 1-3 Alkyl or halogen; R e The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens; m is 0. The compound as claimed in claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, The compound represented by formula (I) has the structure shown in formula (IA) or formula (IB): Where R 51 R 62 Each independently is hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-2 Alkylene-R7, the C 1-3 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl; R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens; n is 1; s is 1, 2, or 3; t is 0, 1, or 2; And when s is 1, t is not 0; R0 is independently hydrogen and C. 1-8 Alkyl groups, halogens; q can be 0, 1, 2, or 3; p is 0, 1, or 2. The compound as claimed in claim 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R1 is fluorine; R2 and R3 are each hydrogen independently. The compound as claimed in claim 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R1 is hydrogen; R2 is hydrogen; R3 is C. 1-3 alkyl. The compound as claimed in claim 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R 51 Groups selected from the following group: -CH2-CN, -(CH2)2-S(O)2-CH3, -(CH2)2-S(O)2-NH2, methyl, -CH2-CF3, -CH(CF3)-CH3, -CH2-CHF2, -CH2-C(O)NH2, -(CH2) 0-1 -Halocyclopropyl (preferably -(CH2)) 0-1 -Fluorocyclopropyl), -(CH2) 0-1 -Halogenated cyclobutyl (preferably -(CH2)) 0-1 -Fluorocyclobutyl), -(CH2) 0-1 -Halocyclopentyl (preferably -(CH2)) 0-1 -Fluorocyclopentyl), -(CH2) 0-1 -Halogenated cyclohexyl (preferably -(CH2)) 0-1 -Fluorocyclohexyl), -(CH2) 0-1 -Cyclopropyl, -(CH2) 0-1 -cyclobutyl, -(CH2) 0-1 -cyclopentyl, -(CH2) 0-1 - Cyclohexyl, azirhexacyclobutyl, oxacyclobutyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, oxazolidinyl, dioxopranyl, piperidinyl, piperazine, morpholinyl, dioxohexacycloyl, thiomorpholinyl, thiomorpholin-1,1-dioxide, tetrahydropyranyl. The compound as claimed in claim 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R 62 It is hydrogen. The compound as claimed in claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, Compounds of formula (I) are selected from any of the following structures: A compound of formula (A), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: in, T is a linker group containing a functional group that can react with a thiol group; L is a linking group containing either non-cleavable or cleavable units; R1 is hydrogen or halogen; R2 and R3 are each independently hydrogen or C. 1-8 Alkyl (preferably C) 1-6 Alkyl, more preferably C 1-3 alkyl); Furthermore, R1, R2, and R3 are not all hydrogen at the same time; n is 1, 2, or 3; R 41 R 42 R 43 R 44 Each can be either hydrogen or halogen; R5, R a R b R c R d R e R f m is selected from the following group of definitions: (i) R5 is hydrogen, C 1-8 Alkyl, C 3-6 cycloalkyl, -C 1-4 Alkylene-C 3-6 cycloalkyl, halogenated C 1-8 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-4 Alkylene-R7, the C 1-8 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-8 alkyl; R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-4 Alkyl, -S(O)-C 1-4 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens; R a R b R c R d R e R f Each independently is hydrogen, C 1-8 Alkyl or halogen; m is 0, 1, or 2; (ii)R a R b R d R e R f Each independently is hydrogen, C 1-8 Alkyl or halogen; R c The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens; m is 0, 1, or 2; (iii)R a R b R d R c R f Each independently is hydrogen, C 1-8 Alkyl or halogen; R e The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens; m can be 0, 1, or 2. The compound of claim 14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, The compound represented by formula (A) has the structure shown in formula (AA) or formula (AB): Where R 51 For hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-2 Alkylene-R7, the C 1-3 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl; R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens; n is 1; s is 1, 2, or 3; t is 0, 1, or 2; And when s is 1, t is not 0; R0 is independently hydrogen and C. 1-8 Alkyl groups, halogens; q can be 0, 1, 2, or 3; p is 0, 1, or 2. The compound of claim 14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, T is T1-T2-; where T1 is a functional group selected from the following group: Where X is a halogen; T2 is absent or is a linking group selected from the following group: The waveform line represents the connection point with T1, the asterisk represents the connection point with L, and t1 is an integer from 1 to 10; t2 and t3 are each an integer from 0 to 10 independently. The compound of claim 14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, T is selected from The asterisk indicates the connection point with L. A compound of formula (B), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: Where Ab represents an antibody or antigen-binding fragment; y is any value between 1 and 10; S represents the sulfur atom of the thiol group in the Ab portion; T1' is selected from: The wavy line indicates the connection point with T2, and the asterisk indicates the connection point with S; T2 is absent or is a linking group selected from the following groups: The wavy line represents the connection point with T1', the asterisk represents the connection point with L, and t1 is an integer from 1 to 10; t2 and t3 are each an independent integer from 0 to 10. L is a linking group containing either non-cleavable or cleavable units; R5, R a R b R c R d R e R f m, R 41 R 42 R 43 R 44 R1, R2, R3, R1, and n are as defined in claim 14. The compound of claim 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, T1' is selected from The wavy line represents the connection point with T2, and the asterisk represents the connection point with S. The compound of claim 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, The compound represented by formula (B) has the structure shown in formula (BA) or formula (BB): Where R 51 For hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl, 4- to 6-membered heterocyclic alkyl or -C 1-2 Alkylene-R7, the C 1-3 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl; R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens; n is 1; s is 1, 2, or 3; t is 0, 1, or 2; And when s is 1, t is not 0; R0 is independently hydrogen and C. 1-8 Alkyl groups, halogens; q can be 0, 1, 2, or 3; p is 0, 1, or 2. The compound of claim 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, T1'-T2 are selected from The wavy line represents the connection point with L, and the asterisk represents the connection point with S. The compound as claimed in claim 14 or 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, L is L1-L2-L3; where L1 either does not exist or is a structure selected from the following group: The wavy line represents the connection point with L2, the asterisk represents the connection point with T or T2, and q1 is an integer from 1 to 8; R L1 R L2 Each of the following is an independent structure selected from the group below: Where p1 is an integer from 1 to 12; u is an integer from 1 to 15; s1, s2, s3, and s4 are each an integer from 1 to 15 independently; and v is an integer from 1 to 20. R 1 R 2 Each independently is C 1-6 alkyl; W does not exist or is -NH-C 1-3 alkylene-; Ring A is a benzene ring or a 4- to 6-membered heterocyclic alkyl ring; L2 is a short peptide composed of 2 to 4 amino acid residues; the amino acid residues are selected from natural amino acid residues and non-natural amino acid residues; L3 either does not exist or has the following structure: Where R L3 For hydrogen or The wavy line represents the connection point with -N-, and the asterisk represents the connection point with L2. The compound as claimed in claim 22, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, L2 is a dipeptide or tripeptide; preferably, L2 is a dipeptide or tripeptide selected from the group consisting of: phenylalanine-lysine, valine-alanine, valine-lysine, alanine-lysine, valine-citrulline, phenylalanine-citrulline, leucine-citrulline, isoleucine-citrulline, phenylalanine-arginine, tryptophan-citrulline, glutamic acid-valine-alanine, glutamic acid-valine-citrulline, glycine-valine-citrulline, alanine-alanine-alanine, and alanine-alanine-asparagine. The compound as claimed in claim 22, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, L3 is selected from The wavy line represents the connection point with -N-, and the asterisk represents the connection point with L2. The compound of claim 14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, TL is a structure selected from the following formula: Where R L1 R L2 L2 is defined as in claim 22, and t1 is an integer from 1 to 10. The compound of claim 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, T1'-T2-L is a structure selected from the following formula: Where R L1 R L2 L2, t1 are defined as in claim 22; wherein the waveform line represents the connection point with -N-, and the asterisk represents the connection point with S. The compound of claim 14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, TL is selected from the structure in Table A. The compound of claim 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, T1'-T2-L is selected from the structure in Table B. The compound of claim 14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, TL is selected from the structure of Table C. The compound of claim 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, T1'-T2-L is selected from the structure of Table D. The compound of claim 14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, The compound shown in formula (A) is selected from the structures in Table E. The compound of claim 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, The compound shown in formula (B) is selected from the structures in Table F, where Ab is an antibody against Her 2 or its antigen-binding fragment; y is any value between 1 and 10. The compound as claimed in claim 14 or 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R1 is a halogen; R2 and R3 are each hydrogen independently. The compound as claimed in claim 14 or 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R1 is fluorine; R2 and R3 are each hydrogen independently. The compound as claimed in claim 14 or 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R1 is hydrogen; R2 is hydrogen; R3 is C. 1-3 alkyl. The compound as claimed in claim 14 or 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R5 is hydrogen, C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl (preferably fluorinated C) 1-3 Alkyl), 4- to 6-membered heterocyclic alkyl or -C 1-2 Alkylene-R7, the C 1-3 Alkyl, C 3-6 Cycloalkyl groups, 4- to 6-membered heterocyclic alkyl groups are unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of: halogens, C 1-3 alkyl; R7 is C 3-6 Cycloalkyl, -S(O)2-C 1-3 Alkyl, -S(O)-C 1-3 Alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens; R a R b R c R d R e R f Each independently is hydrogen, C 1-3 Alkyl or halogen; m is 0. The compound as claimed in claim 14 or 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R5 is C 1-3 Alkyl, C 3-6 cycloalkyl, -C 1-2 Alkylene-C 3-6 cycloalkyl, halogenated C 1-3 Alkyl (preferably fluorinated C) 1-3 alkyl) or -C 1-2 Alkylene-R7, the C 3-6 The cycloalkyl group is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of halogens; R7 is -S(O)2-C 1-3 Alkyl groups or -S(O)2-NH2; R c R d R e R f Each is independently hydrogen; m is 0. The compound as claimed in claim 14 or 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, R a R b R d R e R f Each independently is hydrogen, C 1-3 Alkyl or halogen (preferably R) a R b R d R e R f (for hydrogen); R c The saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is formed by connection with R5; the saturated 4- to 7-membered nitrogen-containing heterocyclic alkyl group is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of: C 1-8 Alkyl groups, halogens; m is 0 or 1. The compound as claimed in claim 15 or 20, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, In compounds of formula (AA) or formula (BA), the structure Selected from the following structure: The compound as claimed in claim 15 or 20, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, In compounds of formula (AB) or formula (BB), the structure Selected from the following structure: The compound as claimed in claim 14 or 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, is characterized in that, In the compounds shown in formula (A) or formula (B), the structure Selected from the following structure: A method for preparing the compound of formula (B), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, The method includes coupling Ab with a compound of formula (A); In this context, Ab represents an antibody or antigen-binding fragment. A pharmaceutical composition, characterized in that, include The compound of any one of claims 1-13, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof; or The compound of claim 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof; And pharmaceutically acceptable carriers. Use of a compound of any one of claims 1-13, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a compound of claim 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition of claim 43, in the preparation of a medicament for the treatment or prevention of diseases associated with mutations, expression imbalances, allosteric changes, or functional abnormalities of GSPT1, N-MYC, or C-MYC proteins. The use according to claim 44 is characterized in that, The diseases mentioned are selected from esophageal cancer, brain tumor, lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, urothelial carcinoma, non-Hodgkin's lymphoma, central nervous system tumors, prostate cancer, thyroid cancer, acute myeloid leukemia, and myelodysplastic syndrome. The use of any compound of claims 1-13, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or the compound of claim 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or the pharmaceutical composition of claim 43, in the preparation of a GSPT1 degrading agent. Use of a compound of any one of claims 1-13, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a compound of claim 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition of claim 43, in the preparation of a medicament for treating diseases associated with or caused by GSPT1. The use according to claim 47 is characterized in that, The diseases mentioned are those related to GSPT1, selected from glioma, thyroid cancer, lung cancer, colorectal cancer, head and neck cancer, gastric cancer, liver cancer, pancreatic cancer, kidney cancer, urothelial carcinoma, prostate cancer, testicular cancer, breast cancer, cervical cancer, endometrial cancer, ovarian cancer, melanoma, multiple myeloma, acute myeloid leukemia, and myelodysplastic syndrome.

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