Isoindoline compound containing phenoxyalkylamine structure, pharmaceutical composition comprising same, and use thereof

Abstract

The present invention relates to an isoindoline compound containing a phenoxyalkylamine structure, a pharmaceutical composition comprising same, and a use thereof. The isoindoline compound containing a phenoxyalkylamine structure has a structure as shown in formula (I). The compound of the present invention has increased proliferation inhibitory activity against cancer cells and a degradation effect on proteins such as GSPT1, and has practical value.

Description

Isoindoline compounds containing phenethyloxy alkylene amine structure, pharmaceutical compositions and applications thereof

[0001] This application claims priority to the Chinese patent application No. 202410874928.6, filed on July 1, 2024, entitled "Isoindoline compounds containing phenethyloxy alkylene amine structure, pharmaceutical compositions and applications thereof", the entire contents of which are incorporated herein by reference. TECHNICAL FIELD

[0002] The present application relates to the technical field of medicine, in particular to an isoindoline compound containing a phenethyloxy alkylene amine structure, a pharmaceutical composition thereof and applications thereof. BACKGROUND

[0003] Eukaryotic peptide chain release factor 3a (eRF3a) is encoded by G1 to S phase transition 1 gene (GSPT1) and is up-regulated in various human cancers. It is a potential proto-oncogene and participates in various biological processes as a key member of the peptide chain release factor, including terminating protein translation, regulating intracellular mRNA degradation, regulating cell growth cycle and apoptosis, and participating in cytoskeleton formation. Recent evidence shows that GSPT1 is abnormally highly expressed in various malignant tumors and plays an important role in the occurrence and development of tumors. GSPT1 is a carcinogenic driver factor for various cancer types, including hematological tumors, breast cancer, liver cancer, gastric cancer and prostate cancer, etc. In addition, GSPT1 is significantly up-regulated in cancer tissues and cell lines, its high expression is positively correlated with tumor size, and its overexpression can promote cancer cell proliferation and migration.

[0004] There are many literatures reporting GSPT1 degradation agents with different structures, including WO2023070120A1, WO2023201282A1, WO2023274246A1, WO2023069708A1, etc. The activity and selectivity of existing GSPT1 degradation agents need to be further improved. Therefore, it is of great research value and practical significance to develop GSPT1 degradation agents with higher activity and selectivity, further improve the treatment effect of tumors and reduce the toxic side effects of drugs. SUMMARY

[0005] The purpose of the present application is to provide a class of isoindoline compounds containing phenethyloxy alkylene amine structure with novel structure and pharmaceutical compositions thereof, which are CRBN E3 ligase modulators (cereblon modulators) with improved proliferation inhibition activity on cancer cells and degradation effect on GSPT1 and other proteins.

[0006] The present application provides, in a first aspect, a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0007] wherein,

[0008] R1is hydrogen or halogen;

[0009] R2, R3are each independently hydrogen or C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl);

[0010] and R1, R2, R3are not simultaneously hydrogen;

[0011] n is 1, 2 or 3;

[0012] R 41 , R 42 , R 43 , R 44 are each independently hydrogen or halogen;

[0013] R5, R6, R a , R b , R c , R d , R e , R f , m is selected from the following definitions:

[0014] (i) R5, R6are each independently 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 heterocycloalkyl or -C 1-4 alkylene-R7, said C 1-8 alkyl, C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl are unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen, C 1-8 alkyl;

[0015] R7is C 3-6 cycloalkyl, -S(O)2-C 1-4 alkyl, -S(O)-C 1-4 alkyl, -S(O)2-NH2, -C(O)NH2, hydroxy or cyano; said C 3-6 cycloalkyl are unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen;

[0016] R a , Rb , R c , R d , R e , R f each independently hydrogen, C 1-8 alkyl or halogen;

[0017] m is 0, 1 or 2;

[0018] (ii) R6is hydrogen, C 1-8 alkyl, C 3-6 cycloalkyl, -C 1-4 alkylene-C 3-6 cycloalkyl, haloC 1-8 alkyl, or 4- to 6-membered heterocycloalkyl, which C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen, C 1-8 alkyl;

[0019] R a , R b , R d , R e , R f each independently hydrogen, C 1-8 alkyl or halogen;

[0020] R c and R5are joined to form a saturated 4- to 7-membered nitrogen-containing heterocycloalkyl; which saturated 4- to 7-membered nitrogen-containing heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of C 1-8 alkyl, halogen;

[0021] m is 0, 1 or 2;

[0022] (iii) R6is hydrogen, C 1-8 alkyl, C 3-6 cycloalkyl, -C 1-4 alkylene-C 3-6 cycloalkyl, haloC 1-8 alkyl, or 4- to 6-membered heterocycloalkyl, which C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen, C 1-8 alkyl;

[0023] R a , R b , R d , R c , R f each independently hydrogen, C 1-8 alkyl or halogen;

[0024] R eR5is connected to form a saturated 4- to 7-membered nitrogen-containing heterocycloalkyl; said saturated 4- to 7-membered nitrogen-containing heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of C 1-8 alkyl, halogen;

[0025] m is 0, 1 or 2.

[0026] In some embodiments, R1is halogen; R2, R3are each independently hydrogen.

[0027] In some embodiments, R1is halogen; R2, R3are each independently hydrogen.

[0028] In some embodiments, R1is hydrogen; R2is hydrogen; R3is C 1-3 alkyl.

[0029] In some embodiments, R5is hydrogen, C 1-3 alkyl, C 3-6 ycloalkyl, -C 1-2 alkylene-C 3-6 ycloalkyl, halogenated C 1-3 alkyl (preferably fluorinated C 1-3 alkyl), 4- to 6-membered heterocycloalkyl or -C 1-2 alkylene-R7, said C 1-3 alkyl, C 3-6 ycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen, C 1-3 alkyl;

[0030] R6is hydrogen;

[0031] R7is C 3-6 ycloalkyl, -S(O)2-C 1-3 alkyl, -S(O)-C 1-3 alkyl, -S(O)2-NH2, -C(O)NH2, hydroxyl or cyano; said C 3-6 ycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen;

[0032] R a , R b , R c , R d , R e , R f each independently hydrogen, C 1-3 alkyl or halogen;

[0033] m is 0.

[0034] In some embodiments, R6is hydrogen, C 1-3 alkyl, C 3-6cycloalkyl, -C 1-2 alkylene-C 3-6 cycloalkyl, haloC 1-3 alkyl, or 4- to 6-membered heterocycloalkyl, said C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of halogen, C 1-3 alkyl;

[0035] R a , R b , R d , R e , R f each independently is hydrogen, C 1-3 alkyl or halogen;

[0036] R c and R5join to form a saturated 4- to 7-membered nitrogen-containing heterocycloalkyl; said saturated 4- to 7-membered nitrogen-containing heterocycloalkyl is unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of C 1-8 alkyl, halogen;

[0037] m is 0 or 1.

[0038] In some embodiments, R6is hydrogen, C 1-3 alkyl, C 3-6 cycloalkyl, -C 1-2 alkylene-C 3-6 cycloalkyl, haloC 1-3 alkyl, or 4- to 6-membered heterocycloalkyl, said C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of halogen, C 1-3 alkyl;

[0039] R a , R b , R d , R c , R f each independently is hydrogen, C 1-3 alkyl or halogen;

[0040] R e and R5join to form a saturated 4- to 7-membered nitrogen-containing heterocycloalkyl; said saturated 4- to 7-membered nitrogen-containing heterocycloalkyl is unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of C 1-8 alkyl, halogen;

[0041] m is 0.

[0042] In some embodiments, the 4- to 6-membered heterocycloalkyl group is selected from the group consisting of azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, oxazolidinyl, dioxolanyl, piperidinyl, piperazinyl, morpholinyl, dioxanyl, thiomorpholinyl, thiomorpholin-1,1 -dioxide, tetrahydropyranyl, pyrrolidin-2-one, dihydrofuran-2(3H)-one, morpholin-3-one, piperazin-2-one, and piperidin-2-one.

[0043] In some embodiments, the saturated 4- to 7-membered nitrogen-containing heterocycloalkyl group is selected from the group consisting of azetidinyl, tetrahydropyrrolyl, oxazolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiomorpholin-1,1 -dioxide, pyrrolidin-2-one, morpholin-3-one, piperazin-2-one, and piperidin-2-one.

[0044] In some embodiments, the compound of Formula (I) is of the structure of Formula (I-A) or Formula (I-B):

[0045] wherein R 51 , R 62 each independently is hydrogen, C 1-3 alkyl, C 3-6 cycloalkyl, -C 1-2 alkylene-C 3-6 cycloalkyl, haloC 1-3 alkyl, 4- to 6-membered heterocycloalkyl, or -C 1-2 alkylene-R7, said C 1-3 alkyl, C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of halogen, C 1-3 alkyl;

[0046] R7is 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; said C 3-6 cycloalkyl is unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of halogen;

[0047] n is 1 ;

[0048] s is 1, 2, or 3;

[0049] t is 0, 1, or 2;

[0050] and when s is 1, t is not 0;

[0051] R0each independently is hydrogen, C1-8 alkyl, halo;

[0052] q is 0, 1, 2, or 3;

[0053] p is 0, 1, or 2.

[0054] In some embodiments, R1is hydrogen; R2, R3are each independently hydrogen.

[0055] In some embodiments, R1is hydrogen; R2is hydrogen; R3is C 1-3 alkyl.

[0056] In some embodiments, R 51 is selected from the group consisting of -CH2-CN, -(CH2)2-S(O)2-CH3, -(CH2)2-OH, methyl, ethyl, 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 halocyclobutyl (preferably -(CH2) 0-1 fluorocyclobutyl), -(CH2) 0-1 halocyclopentyl (preferably -(CH2) 0-1 fluorocyclopentyl), -(CH2) 0-1 halocyclohexyl (preferably -(CH2) 0-1 fluorocyclohexyl), -(CH2) 0-1 cyclopropyl, -(CH2) 0-1 cyclobutyl, -(CH2) 0-1 cyclopentyl, -(CH2) 0-1 cyclohexyl, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, oxazolidinyl, dioxolanyl, piperidinyl, piperazinyl, morpholinyl, dioxanyl, thiomorpholinyl, thiomorpholine-1,1-dioxide, tetrahydropyranyl.

[0057] In some embodiments, R 62 is hydrogen.

[0058] In some embodiments, in formula (I-A), R 51 is C 3-6 cycloalkyl or haloC 1-3 alkyl, the C 3-6 cycloalkyl is unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of halo; n is 1.

[0059] In some embodiments, in formula (I-A), R 51 is selected from the group consisting of -CH2-CF3, -CH2-CHF2, cyclopropyl, fluorocyclobutyl, -CH(CF3)-CH3.

[0060] In some embodiments, the compound of formula (I) is selected from any one of the following structures:

[0061] In some embodiments, the compound of formula (I) is selected from any one of the following structures:

[0062] The compound of the first aspect of the present application, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof has significant protein modulation and improved cell proliferation inhibition effect on the proteins including GSPT1, N-MYC and C-MYC, and is a protein modulator to restore protein homeostasis including GSPT1 activity.

[0063] The second aspect of the present application provides a compound represented by formula (II), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0064] wherein R1' is hydrogen or halogen;

[0065] R2', R3' are each independently hydrogen or C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl);

[0066] n is 1, 2 or 3;

[0067] R 41 , R 42 , R 43 , R 44 are each independently hydrogen or halogen;

[0068] R5', R6, R a , R b , R c , R d , R e , R f , m is selected from the following definitions:

[0069] (i) R5' is C 3-6 cycloalkyl, -C 1-4 alkylene-C 3-6 cycloalkyl, halo-C 1-8 alkyl, 4- to 6-membered heterocycloalkyl or -C1-4 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;

[0070] 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;

[0071] 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;

[0072] R a R b R c R d R e R f Each independently is hydrogen, C 1-8 Alkyl or halogen;

[0073] m is 0, 1, or 2;

[0074] (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;

[0075] R a R b R d R e R f Each independently is hydrogen, C1-8 alkyl or halogen;

[0076] R c to form, together with R5', a saturated 4- to 7-membered nitrogen-containing heterocycloalkyl; said saturated 4- to 7-membered nitrogen-containing heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of C 1-8 alkyl, halogen;

[0077] m is 0, 1 or 2;

[0078] (iii) R6is 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 heterocycloalkyl, said C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen, C 1-8 alkyl;

[0079] R a , R b , R d , R c , R f each independently hydrogen, C 1-8 alkyl or halogen;

[0080] R e to form, together with R5', a saturated 4- to 7-membered nitrogen-containing heterocycloalkyl; said saturated 4- to 7-membered nitrogen-containing heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of C 1-8 alkyl, halogen;

[0081] m is 0, 1 or 2.

[0082] In some embodiments, R1', R2', R3' are hydrogen.

[0083] In some embodiments, R5' is C 3-6 cycloalkyl, -C 1-4 alkylene-C 3-6 cycloalkyl, halogenated C 1-8 alkyl, 4- to 6-membered heterocycloalkyl or -C 1-4 alkylene-R7, said C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen, C 1-8 alkyl;

[0084] R6is hydrogen;

[0085] R7is C 3-6cycloalkyl, -S(O)2-C 1-4 alkyl, -S(O)-C 1-4 alkyl, -S(O)2-NH2, -C(O)NH2, hydroxy, or cyano; said C 3-6 cycloalkyl is unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of halogen;

[0086] R a , R b , R c , R d , R e , R f each independently is hydrogen, C 1-8 alkyl, or halogen;

[0087] m is 0, 1, or 2.

[0088] In some embodiments, R6is hydrogen.

[0089] In some embodiments, the compound of Formula (II) is of the structure of Formula (II-A) or Formula (II-B):

[0090] wherein R 51 ' is C 3-6 cycloalkyl, -C 1-2 alkylene-C 3-6 cycloalkyl, haloC 1-3 alkyl, 4- to 6-membered heterocycloalkyl, or -C 1-2 alkylene-R7, said C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of halogen, C 1-3 alkyl;

[0091] R 62 each independently is hydrogen, C 1-3 alkyl, C 3-6 cycloalkyl, -C 1-2 alkylene-C 3-6 cycloalkyl, haloC 1-3 alkyl, 4- to 6-membered heterocycloalkyl, or -C 1-2 alkylene-R7, said C 1-3 alkyl, C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted with 1, 2, or 3 substituents selected from the group consisting of halogen, C 1-3 alkyl;

[0092] R7is C 3-6 cycloalkyl, -S(O)2-C 1-3 alkyl, -S(O)-C 1-3alkyl, -S(O)2-NH2, -C(O)NH2, hydroxy, or cyano; said C 3-6 cycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen;

[0093] n is 1 ;

[0094] s is 1, 2 or 3;

[0095] t is 0, 1 or 2;

[0096] and when s is 1, t is not 0;

[0097] R° is each independently hydrogen, C 1-8 alkyl, halogen;

[0098] q is 0, 1, 2 or 3;

[0099] p is 0, 1 or 2.

[0100] In some embodiments, R 51 is selected from the group consisting of -CH2-CN, -(CH2)2-S(O)2-CH3, -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 halocyclobutyl (preferably -(CH2) 0-1 fluorocyclobutyl), -(CH2) 0-1 halocyclopentyl (preferably -(CH2) 0-1 fluorocyclopentyl), -(CH2) 0-1 halocyclohexyl (preferably -(CH2) 0-1 fluorocyclohexyl), -(CH2) 0-1 cyclopropyl, -(CH2) 0-1 cyclobutyl, -(CH2) 0-1 cyclopentyl, -(CH2) 0-1 cyclohexyl, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, oxazolidinyl, dioxolanyl, piperidinyl, piperazinyl, morpholinyl, dioxanyl, thiomorpholinyl, thiomorpholine-1,1 -dioxide, tetrahydropyranyl.

[0101] In some embodiments, R 62 is hydrogen.

[0102] In some embodiments, in formula (II-A), R 51 is C3-6 cycloalkyl or haloC 1-3 alkyl, said C 3-6 cycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen; n is 1.

[0103] In some embodiments, in formula (II-A), R 51 is selected from the group consisting of -CH2-CF3, -CH2-CHF2, cyclopropyl, fluorocyclobutyl, -CH(CF3)-CH3.

[0104] The third aspect of the present application provides a pharmaceutical composition comprising the compound of the first or second aspect of the present application, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and a pharmaceutically acceptable carrier.

[0105] The fourth aspect of the present application provides use of the compound of the first or second aspect of the present application, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or the pharmaceutical composition of the third aspect of the present application in the preparation of a GSPT1 degrader.

[0106] The fifth aspect of the present application provides use of the compound of the first or second aspect of the present application, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or the pharmaceutical composition of the third aspect of the present application in the preparation of a medicament for treating or preventing a disease associated with GSPT1, N-MYC or C-MYC protein mutation, expression imbalance, allosteric and functional abnormality.

[0107] In some embodiments, the disease includes, but is 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 cancer, non-Hodgkin's lymphoma, central nervous system tumor, prostate cancer, thyroid cancer, acute myeloid leukemia, myelodysplastic syndrome.

[0108] The sixth aspect of the present application provides use of the compound of the first or second aspect of the present application, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or the pharmaceutical composition of the third aspect of the present application in the preparation of a medicament for treating a disease associated with or caused by GSPT1, in particular, treating a cancer associated with GSPT1, such as glioma, thyroid cancer, lung cancer, colorectal cancer, head and neck cancer, gastric cancer, liver cancer, pancreatic cancer, kidney cancer, urothelial cancer, prostate cancer, testicular cancer, breast cancer, cervical cancer, endometrial cancer, ovarian cancer, melanoma, multiple myeloma, acute myeloid leukemia, myelodysplastic syndrome.

[0109] In some embodiments, the compound of the first or second aspect of the present application, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or the pharmaceutical composition of the third aspect of the present application is used for treating solid tumors, in particular for treating breast cancer.

[0110] The seventh aspect of the present application provides a method for degrading GSPT1 protein in a patient in need thereof, comprising administering to the patient the compound of the first or second aspect of the present application, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or the pharmaceutical composition of the third aspect of the present application.

[0111] It should be understood that, within the scope of the present application, each of the technical features described above and in the following (such as the examples) can be combined with each other to form new or preferred technical solutions. Due to the limited space, they will not be listed one by one here. DETAILED DESCRIPTION

[0112] The inventors have made an unexpected discovery that the isoindoline compounds of this type containing phenethyloxy alkylene amine structure can regulate the level of proteins such as GSPT1, have obviously improved inhibitory proliferation effect on tumor cells such as human breast ductal carcinoma cells, and can effectively alleviate or treat cancer and other related diseases. On this basis, the inventors have completed the present application.

[0113] Definitions of terms

[0114] In order to more clearly understand the technical content of the present application, the terms of the present application are further described below.

[0115] "Alkyl" refers to straight-chain and branched saturated aliphatic hydrocarbon groups. "C 1-8 "Alkyl" refers to straight-chain and branched saturated aliphatic hydrocarbon groups. "C 1-6 "Alkyl" refers to straight-chain and branched saturated aliphatic hydrocarbon groups. "C 1-3alkyl; non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, s-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 various branched isomers thereof.

[0116] "Cycloalkyl" and "cycloalkyl ring" are used interchangeably and refer to a saturated monocyclic, bicyclic, or polycyclic cyclic hydrocarbon radical which can be fused to an aryl or heteroaryl group. The cycloalkyl ring can be optionally substituted. In certain embodiments, the cycloalkyl ring contains one or more carbonyl groups, e.g., oxo groups. "C 3-8 "Cycloalkyl" and "cycloalkyl ring" are used interchangeably and refer to a saturated monocyclic, bicyclic, or polycyclic cyclic hydrocarbon radical which can be fused to an aryl or heteroaryl group. The cycloalkyl ring can be optionally substituted. In certain embodiments, the cycloalkyl ring contains one or more carbonyl groups, e.g., oxo groups. "C 3-6 "Cycloalkyl" and "cycloalkyl ring" are used interchangeably and refer to a saturated monocyclic, bicyclic, or polycyclic cyclic hydrocarbon radical which can be fused to an aryl or heteroaryl group. The cycloalkyl ring can be optionally substituted. In certain embodiments, the cycloalkyl ring contains one or more carbonyl groups, e.g., oxo groups. "C

[0117] "Heteroaralkyl" refers to an alkyl group substituted with a heteroaryl group. The point of attachment is through the alkyl group. "Heteroaralkyl" and "heteroaralkyl ring" are used interchangeably and refer to a heteroaryl group as defined herein, which is substituted with an alkyl group. The heteroaralkyl ring can be optionally substituted. In certain embodiments, the heteroaralkyl ring contains one or more carbonyl or thiocarbonyl groups, for example, oxo and thioxo containing groups. "3- to 8-membered heteroaralkyl" refers to a heteroaryl group having 3 to 8 ring atoms, preferably 4 to 8 ring atoms, wherein 1, 2, or 3 ring atoms are heteroatoms selected from nitrogen, oxygen, and sulfur. More preferably, 3- to 6-membered heteroaralkyl refers to a heteroaryl group having 3 to 6 ring atoms, wherein 1 or 2 ring atoms are heteroatoms selected from nitrogen, oxygen, and sulfur. More preferably, 4- to 6-membered heteroaralkyl refers to a heteroaryl group having 4 to 6 ring atoms, wherein 1 or 2 ring atoms are heteroatoms selected from nitrogen, oxygen, and sulfur. Non-limiting examples include pyridylalkyl, pyrimidinylalkyl, pyrazinylalkyl, pyridazinylalkyl, thienylalkyl, furanylalkyl, pyrrolylalkyl, imidazolylalkyl, oxazolylalkyl, thiazolylalkyl, isoxazolylalkyl, isothiazolylalkyl, oxadiazolylalkyl, thiadiazolylalkyl, triazolylalkyl, tetrazolylalkyl, and the like.

[0118] "Heteroaryl" and "heteroaryl ring" are used interchangeably and refer to a monocyclic, bicyclic, or polycyclic 4n+2 aromatic ring system (e.g., having 6 or 10 p-electrons shared in a cyclic array) having ring carbon atoms and ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur. The heteroaryl ring can be optionally substituted. "5- to 10-membered heteroaryl" refers to a monocyclic or bicyclic heteroaryl group having 5 to 10 ring atoms, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms. "5- to 6-membered heteroaryl" refers to a monocyclic heteroaryl group having 5 to 6 ring atoms, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms, non-limiting examples of which include thienyl, furanyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, pyrrolyl, 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, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, tetrazinyl. "9- or 10-membered heteroaryl" refers to a bicyclic heteroaryl group having 9 or 10 ring atoms, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms, non-limiting examples of which include indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzoimidazolyl, benzoxazolyl, benzoisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indolizinyl, purinyl, pyrido[3,2-d]pyrimidinyl, pyrido[2,3-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrido[4,3-d]pyrimidinyl, 1,8-naphthyridinyl, 1,7-naphthyridinyl, 1,6-naphthyridinyl, 1,5-naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. "Heteroatom" refers to nitrogen, oxygen, or sulfur. In heteroaryl groups containing one or more nitrogen atoms, the point of attachment can be a carbon or a nitrogen atom, as valency permits. Heteroaryl bicyclic systems can include one or more heteroatoms in one or both rings.

[0119] "Alkoxy" refers to -O-alkyl, wherein alkyl is as defined above. Preferred C 1-8 "Alkoxy" refers to -O-alkyl, wherein alkyl is as defined above. Preferred C 1-6 "Alkoxy" refers to -O-alkyl, wherein alkyl is as defined above. Preferred C 1-3 "Alkoxy" refers to -O-alkyl, wherein alkyl is as defined above. Preferred C

[0120] "A bond" refers to a covalent bond between two groups attached thereto.

[0121] "Halogen" refers to fluorine, chlorine, bromine, or iodine.

[0122] "Halo" means one or more (e.g., 1, 2, 3, 4, or 5) hydrogens on a group are replaced with a halogen.

[0123] For example, "haloalkyl" means an alkyl group, as previously described, wherein one or more (e.g., 1, 2, 3, 4, or 5) hydrogens are replaced with a halogen. Preferably, haloalkyl is haloC 1-8 alkyl, more preferably haloC 1-6 alkyl, more preferably haloC 1-3 alkyl. Examples of haloalkyl include, but are not limited to, chloromethyl, dichloromethyl, trichloromethyl, chloroethyl, 1,2-dichloroethyl, trichloroethyl, bromoethyl, fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl, trifluoroethyl, and the like.

[0124] For example, "haloalkyl" means an alkyl group, as previously described, wherein one or more (e.g., 1, 2, 3, 4, or 5) hydrogens are replaced with a halogen. Preferably, haloalkyl is haloC 1-8 alkyl, more preferably haloC 1-6 alkyl, more preferably haloC 1-3 alkyl. Examples of haloalkyl include, but are not limited to, chloromethyl, dichloromethyl, trichloromethyl, chloroethyl, 1,2-dichloroethyl, trichloroethyl, bromoethyl, fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl, trifluoroethyl, and the like.

[0125] For example, "haloalkyl" means an alkyl group, as previously described, wherein one or more (e.g., 1, 2, 3, 4, or 5) hydrogens are replaced with a halogen. Preferably, haloalkyl is haloC 3-8 alkyl, more preferably haloC 3-6 alkyl. Examples of haloalkyl include, but are not limited to, chloromethyl, dichloromethyl, trichloromethyl, chloroethyl, 1,2-dichloroethyl, trichloroethyl, bromoethyl, fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl, trifluoroethyl, and the like.

[0126] For example, "haloalkyl" means an alkyl group, as previously described, wherein one or more (e.g., 1, 2, 3, 4, or 5) hydrogens are replaced with a halogen. Preferably, haloalkyl is haloC 1-8 alkyl, more preferably haloC 1-6 alkyl, more preferably haloC 1-3 alkyl. Examples of haloalkyl include, but are not limited to, chloromethyl, dichloromethyl, trichloromethyl, chloroethyl, 1,2-dichloroethyl, trichloroethyl, bromoethyl, fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl, trifluoroethyl, and the like.

[0127] "Amino" means NH2, "cyano" means CN, "nitro" means NO2, "benzyl" means -CH2-phenyl, "oxo" means =O, "carboxy" means -C(O)OH, "acetyl" means -C(O)CH3, "hydroxymethyl" means -CH2OH, "hydroxyethyl" means -CH2CH2OH or -CHOHCH3, "hydroxy" means -OH, "mercapto" means SH, "cyclopropylidene" has the structure:

[0128] "Substituted" means that one or more hydrogen atoms, preferably 1 to 5 hydrogen atoms, of a group are independently of each other replaced by a corresponding number of substituents, more preferably 1 to 3 hydrogen atoms are independently of each other replaced by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, which can be determined (experimentally or theoretically) by the person skilled in the art without undue effort, as to whether a substitution is possible or not. For example, an amino or hydroxy group with a free hydrogen can be unstable when bound to a carbon atom with an unsaturated (e.g. olefinic) bond.

[0129] Unless defined otherwise, "substituents independently of each other selected from" as used herein means that when more than one hydrogen of a group is replaced by a substituent, the kind of substituents selected can be the same or different, each independently of the other.

[0130] Unless defined otherwise, "the same or different, and each independently" as used herein means that when more than one identical substituent group is present in a general formula, the groups can be the same or different, each independently of the other. For example, L is (CR 01 R 02 ) s , when s is 2, i.e. L is (CR 01 R 02 )-(CR 01 R 02 ), wherein the two R 01 groups can be the same or different, the two R 02 groups can be the same or different, each independently of the other, e.g. L can be C(CH3)(CN)-C(CH2CH3)(OH), C(CH3)(CN)-C(CH3)(OH) or C(CN)(CH2CH3)-C(OH)(CH2CH3).

[0131] Unless defined otherwise, any group herein can be substituted or unsubstituted. When the above groups are substituted, the substituents are preferably 1 to 5 of the following 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 C1-6 alkoxy, more preferably C 1-3 halo-C 1-8 alkyl (preferably halo-C 1-6 alkyl, more preferably halo-C 1-3 alkyl), C 3-8 cycloalkyl (preferably C 3-6 cycloalkyl), halo-C 1-8 alkoxy (preferably halo-C 1-6 alkoxy, more preferably halo-C 1-3 alkoxy), C 1-8 alkyl-substituted amino, halo-C 1-8 alkyl-substituted amino, acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxy, 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 (preferably C 2-6 alkynyl, more preferably C 2-4 alkynyl), -CONR a0 R b0 , -C(O)OC 1-10 alkyl (preferably -C(O)OC 1-6 alkyl, more preferably -C(O)OC 1-3 alkyl), -CHO, -OC(O)C 1-10 alkyl (preferably -OC(O)C 1-6 alkyl, more preferably -OC(O)C 1-3 alkyl), -SO2C 1-10 alkyl (preferably -SO2C 1-6 alkyl, more preferably -SO2C 1-3 alkyl), -SO2C 6-10 aryl (preferably -SO2C6aryl, such as -SO2-phenyl), -COC 6-10 aryl (preferably -COC6aryl, such as -CO-phenyl), 4- to 6-membered saturated or unsaturated mono-heterocyclic ring, 4- to 6-membered saturated or unsaturated monocyclic ring, 5- to 6-membered monocyclic heteroaryl ring, 8- to 10-membered bicyclic heteroaryl ring, spirocyclic ring, spiroheterocyclic ring, bridged cyclic ring, or bridged heterocyclic ring, wherein R a0 , R b0 each independently is hydrogen or C 1-3 alkyl.

[0132] In the present invention, when two or more "preferably" appear in one scheme, any two "preferably" can be independent of each other.

[0133] In the present application, when the number of substituents is more than one, any two substituents can be the same or different. For example, they can be two same or different halogen substitutions, one halogen and one hydroxyl substitution.

[0134] Each of the above-mentioned substituents can be further substituted by the groups described herein.

[0135] When the 4- to 6-membered saturated monocyclic heterocycle described herein is substituted, the positions of the substituents can be at their possible chemical positions, and exemplary representative substitution cases of the monocyclic heterocycle are shown below:

[0136] wherein "Sub" represents each of the above-mentioned substituents; represents the position of connection with other atoms.

[0137] Pharmaceutical composition

[0138] Generally, the compounds of the present application, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, can be administered in a suitable dosage form prepared by combining the active ingredient with one or more pharmaceutically acceptable carriers. These dosage forms include those suitable for oral, rectal, topical, buccal, and other non-gastrointestinal administration (e.g., subcutaneous, intramuscular, intravenous, etc.). For example, dosage forms suitable for oral administration include capsules, tablets, granules, syrups, and the like. The compounds of the present application contained in these formulations can be in solid or granular form; solutions or suspensions in aqueous or non-aqueous liquids; water-in-oil or oil-in-water emulsions, and the like. The above-mentioned dosage forms can be prepared by conventional pharmaceutical processes using the active compound in combination with one or more carriers or excipients. The above-mentioned carriers need to be compatible with the active compound or other excipients. The active compound can form a solution or suspension with the carrier.

[0139] "Pharmaceutically acceptable carrier" refers to a non-toxic, inert, solid, semi-solid, or liquid material or liquid filling machine, diluent, encapsulating material, or auxiliary formulation or any type of excipient, which is compatible with the patient, preferably a mammal, more preferably a human, and is suitable for delivering the active agent to the target site without terminating the activity of the agent.

[0140] "Compound of the present application" refers to the compound of formula (I) of the present application, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, which has the effect of modulating the level of GSPT1 and other proteins and inhibiting the proliferation of tumor cells, and intermediate compounds for preparing the compound of formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0141] The composition of the present application is formulated, dosed and administered in a manner compatible with good medical practice. The "therapeutically effective amount" of the compound to be administered is dependent on the type of the specific disorder being treated, the individual's size, the severity of the disorder, the manner of administration and the like.

[0142] "Therapeutically effective amount" means an amount of a compound of the present application that will elicit the biological or medical response of a subject, for example, reduction or inhibition of an enzyme or a protein activity, or ameliorate symptoms, arrest or delay disease progression, or prevent a disease, etc.

[0143] "Patient" means an animal, preferably a mammal, more preferably a human. The term "mammal" refers to warm blooded vertebrate mammals, including, for example, cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice, pigs, and humans.

[0144] "Treat" means to reduce, slow the progression of, alleviate, prevent, or maintain 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.

[0145] The "pharmaceutically acceptable salts" include pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. Pharmaceutically acceptable acid addition salts are salts of an acid formed by combination with the free base which retain the biological effectiveness and non-toxicity of the free base. These salts are prepared in a manner known per se.

[0146] "Pharmaceutically acceptable base addition salts" include, but are not limited to, salts of inorganic acids such as sodium, potassium, calcium, and magnesium salts, and the like. Salts of organic acids, including but not limited to, ammonium salts, triethylamine salts, lysine salts, arginine salts, and the like. These salts are prepared in a manner known per se.

[0147] When the compounds of the present application as represented by Formula (I) contain one or more chiral centers, they can exist as different optically active forms. When the compounds of Formula (I) contain one chiral center, the compound comprises a pair of enantiomers. Both enantiomers of the compound and mixtures thereof, such as racemic mixtures, are within the scope of the present application. The enantiomers can be resolved by methods known in the art, such as crystallization and chiral chromatography. When the compounds of Formula (I) contain more than one chiral center, the compound comprises pairs of enantiomers and diastereomers. All enantiomers and diastereomers of the compound, mixtures of enantiomers, mixtures of diastereomers, and mixtures of enantiomers and diastereomers are within the scope of the present application. The enantiomers, diastereomers can be resolved by methods known in the art, such as crystallization and preparative chromatography.

[0148] Methods of preparation

[0149] The present application provides methods for preparing compounds of Formula (I) which can be synthesized using standard synthetic techniques known to those skilled in the art or using methods known in the art in combination with the methods described herein. The solvents, temperatures, and other reaction conditions given herein can be varied by one skilled in the art. The reactions can be used in sequence to provide the compounds of the present application, or they can be used to synthesize fragments which are subsequently incorporated by the methods described herein and / or methods known in the art.

[0150] The compounds described herein can be synthesized using methods analogous to those described below or the exemplary methods described in the examples, or using related disclosures by those skilled in the art, by using appropriate alternative starting materials. The starting materials used to synthesize the compounds described herein can be synthesized or can be obtained from commercial sources. The compounds described herein and other related compounds having different substituents can be synthesized using techniques and starting materials known to those skilled in the art. The general methods for preparing the compounds disclosed herein can be derived from reactions known in the art and the reactions can be modified by reagents and conditions deemed appropriate by those skilled in the art to introduce the various moieties in the molecules provided herein.

[0151] As an example, a general synthetic route for the compounds of Formula (I-A) of the present application is shown in Scheme 1 below:

[0152] Scheme 1

[0153] The compound of Formula (I-A-1) is reacted with the compound of Formula (I-A-2) or a salt thereof in the presence of triphosgene to form the compound of Formula (I-A-a). The reaction can be carried out by reacting the compound of Formula (I-A-1) with triphosgene in an inert solvent at a temperature (e.g., -20 °C to 80 °C, preferably at 0 °C to 60 °C, more preferably at 20 °C to 60 °C) for a period of time (e.g., 5 minutes to 48 hours, preferably 0.1 hour to 5 hours), and then the reaction mixture is reacted with the compound of Formula (I-A-2) or a salt thereof in the presence of a basic reagent at a temperature (e.g., -20 °C to 80 °C, preferably at 0 °C to 60 °C, more preferably at 20 °C to 60 °C) for a period of time (e.g., 5 minutes to 48 hours, preferably 0.1 hour to 5 hours), to form the compound of Formula (I-A-a). The inert solvent and the basic reagent can be known in the art, and the inert solvent can be selected from, for example, C 1-4alkyl alcohol, 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, and the like, or a combination thereof. The basic reagent can be selected from, for example, aqueous 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, imidazole, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, sodium acetate, sodium phenoxide, sodium benzoate, sodium citrate, N-methylmorpholine, pyridine, or a combination thereof. In formula (I-A-1), R p represents a nitrogen protecting group, which can be, for example, a common nitrogen protecting group known in the art, such as benzyloxycarbonyl (-Cbz), tert-butyloxycarbonyl (-Boc).

[0154] The compound of formula (I-A-a) is deprotected under suitable conditions, such as acid conditions or palladium-carbon / hydrogen reduction, to give the compound of formula (I-A).

[0155] As an example, a general synthetic route for the compounds of formula (I-A) of the present application is shown in Scheme 2 below:

[0156] Scheme 2

[0157] The compound of formula (I-A-3) is reacted with the compound of formula (I-A-2) or a salt thereof in the presence of triphosgene to form the compound of formula (I-A-b). The reaction can be carried out in an inert solvent at a temperature (for example, -20 °C to 80 °C, preferably 0 °C to 60 °C, more preferably 20 °C to 60 °C) for a period of time (for example, 5 minutes to 48 hours, preferably 0.1 hour to 5 hours) by reacting the compound of formula (I-A-3) with triphosgene, and then the reaction mixture is reacted with the compound of formula (I-A-2) or a salt thereof in the presence of a basic reagent at a temperature (for example, -20 °C to 80 °C, preferably 0 °C to 60 °C, more preferably 20 °C to 60 °C) for a period of time (for example, 5 minutes to 48 hours, preferably 0.1 hour to 5 hours), thereby obtaining the compound of formula (I-A-b). The inert solvent and the basic reagent can be known in the art, wherein the inert solvent can be selected from, for example, C 1-4alkyl alcohol, 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, and the like, or a combination thereof. The basic reagent can be selected from, for example, aqueous 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, imidazole, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, sodium acetate, sodium phenoxide, sodium benzoate, sodium citrate, N-methylmorpholine, pyridine, or a combination thereof. R p represents a nitrogen protecting group, for example, can be a common nitrogen protecting group known in the art, such as -Cbz, -Boc.

[0158] The compound of formula (I-A-b) is deprotected under suitable conditions, for example, under acidic conditions or under palladium-carbon / hydrogen reduction conditions, to give the compound of formula (I-A-c).

[0159] The compound of formula (I-A-c) is subjected to a reductive amination reaction with a corresponding aldehyde or ketone compound to give the compound of formula (I-A). The reductive amination reaction is carried out by reacting the compound of formula (I-A-c), the aldehyde or ketone compound, and a reducing agent in an inert solvent at a temperature ranging from, for example, -20 °C to 80 °C, preferably from 0 °C to 60 °C, more preferably from 20 °C to 60 °C, for a period of time ranging from, for example, 0.5 hour to 48 hours, preferably from 0.5 hour to 5 hours, to give the compound of formula (I-A). The inert solvent and the reducing agent can be known in the art, wherein the reducing agent can be selected from, for example, tetrabutylammonium borohydride, sodium malonate borohydride, sodium triacetoxyborohydride, sodium cyanoborohydride, sodium borohydride, lithium borohydride, potassium borohydride, borane, and the like. The inert solvent can be selected from, for example, C 1-4 alkyl alcohol, 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, and the like, or a combination thereof.

[0160] As an example, the compound of formula (I-B) of the present application when R 62 The general synthetic route for the compound of formula (I-B-b) when R

[0161] Scheme 3

[0162] The compound of formula (I-B-1) is reacted with the compound of formula (I-A-2) or a salt thereof in the presence of triphosgene to form the compound of formula (I-B-a). The reaction can be carried out by reacting the compound of formula (I-B-1) with triphosgene in an inert solvent at a temperature (e.g., -20 °C to 80 °C, preferably at 0 °C to 60 °C, more preferably at 20 °C to 60 °C) for a period of time (e.g., 5 minutes to 48 hours, preferably 0.1 hour to 5 hours), and then the reaction mixture is reacted with the compound of formula (I-A-2) or a salt thereof in the presence of a basic reagent at a temperature (e.g., -20 °C to 80 °C, preferably at 0 °C to 60 °C, more preferably at 20 °C to 60 °C) for a period of time (e.g., 5 minutes to 48 hours, preferably 0.1 hour to 5 hours) to form the compound of formula (I-B-a). The inert solvent and the basic reagent can be known in the art, and the inert solvent can be selected from the group consisting of C 1-4 alkyl alcohol, 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, and the like, or a combination thereof. The basic reagent can be selected from the group consisting of aqueous 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, imidazole, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, sodium acetate, sodium phenoxide, sodium benzoate, sodium citrate, N-methylmorpholine, pyridine, or a combination thereof. The R p represents a nitrogen protecting group, which can be a common nitrogen protecting group known in the art, such as -Cbz, -Boc.

[0163] The compound of formula (I-B-a) is deprotected to form the compound of formula (I-B-b) under suitable conditions, such as acid conditions or palladium-carbon / hydrogen gas reduction conditions.

[0164] The main advantages of the present application include:

[0165] Compared with the prior art compounds, the isoindoline compounds of the present application containing a phenethyloxyalkyleneamine structure as a GSPT1 and the like protein degrading agent have a significantly improved inhibitory effect on the proliferation of tumor cells, such as human breast ductal carcinoma cells and the like, and the inhibitory activity of the proliferation of MDA-MB-453 cells is IC 50 less than 500 nmol / L (nM), and in some embodiments, has an IC 50 value of less than 300 nM, and in some embodiments, has an IC 50values, in some embodiments have an IC 50 values, in some embodiments have an IC

[0166] The application will be further described in conjunction with specific examples. It should be understood, however, that these examples are intended to illustrate the application and are not intended to limit the scope of the application. The experimental procedures in the following examples, unless otherwise indicated, were carried out in accordance with conventional procedures 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 specified. As used herein, the terms used are intended to have the same meaning as understood by one of ordinary skill in the art. Furthermore, any method and material similar or equivalent to those described herein can be used in the practice of the present application.

[0167] Known starting materials were either synthesized using or according to methods known in the art, or were purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, and Daejung Chemicals.

[0168] Unless otherwise specified, the reactions in the examples were carried out under a nitrogen or argon atmosphere.

[0169] DCM: dichloromethane, DMF: dimethylformamide, THF: tetrahydrofuran, Pd(dppf)Cl2: [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium, Pin2B2: pinacolatoboronate, SEM-Cl: 2-(trimethylsilyl)ethoxymethyl chloride, ACN: acetonitrile, FA: formic acid, n-BuLi: n-butyllithium, KOAc: potassium acetate, HATU: 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate, EDCI: 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride.

[0170] The percentage content involved in the present application, unless otherwise specified, refers to mass percentage for solid-liquid mixing and solid-phase-solid-phase mixing, and refers to volume percentage for liquid-phase-liquid-phase mixing.

[0171] The final product of the present application can be obtained by purification after preparation, and the purification and separation conditions can be obtained by one skilled in the art according to the characteristics of the compound and by routine knowledge in the art.

[0172] As used herein, room temperature refers to about 20-30 °C. Overnight refers to 10-24 h.

[0173] Preparation of intermediate compound V1

[0174] Step 1: Into a 2 L three-necked flask, V1-1 (50.0 g, 231.92 mmol) was added, and stirred under nitrogen protection in an ice-water bath at 0 °C. Borane tetrahydrofuran (1 M, 510.23 mmol, 510.2 mL) was added dropwise slowly. After the addition was completed, the reaction was continued to stir for 12 h. After the reaction was cooled to room temperature, it was stirred in a water bath, and methanol was added dropwise slowly to quench the reaction until no gas bubbles were generated. The reaction was dried under reduced pressure. The obtained crude product was separated and purified by Combi Flash silica gel column (120 g, 0-30 vol% / 100-70 vol% ethyl acetate / petroleum ether) to obtain orange-red solid V1-2 (45.46 g, 97.23% yield). 1 H NMR (400 MHz, CDC13) δ 8.25 (d, J = 2.4 Hz, 1H), 8.07 (dd, J = 8.5, 2.4 Hz, 1H), 7.49 (d, J = 8.4 Hz, 1H), 3.94 (t, J = 6.5 Hz, 2H), 3.11 (t, J = 6.5 Hz, 2H). LC-MS m / z (ESI): 201.9 [M+1] + .

[0175] Step 2: V1-2 (10.09 g, 50.05 mmol) was dissolved in toluene (100.0 mL), and tetrabutylammonium hydrogen sulfate (10.03 g, 29.53 mmol) was added. The mixture was stirred in an ice-water bath at 0 °C, and tert-butyl bromoacetate (39.05 g, 200.19 mmol) was added dropwise slowly. A solution of sodium hydroxide (9.61 g, 240.23 mmol) dissolved in H2O (60.0 mL) was added dropwise slowly. After the addition was completed, the reaction was gradually warmed to room temperature and continued to stir for 12 h. The reaction was poured into a separatory funnel, and the organic phase was separated. The aqueous phase was extracted with ethyl acetate (60 mL), and the combined organic phase was dried with anhydrous sodium sulfate. After filtration, the reaction was dried under reduced pressure. The obtained crude product was dried under reduced pressure with an oil pump to obtain yellow oily liquid V1-3 (15.6 g, 98.72% yield). 1 H NMR (400 MHz, CDC13) δ 8.18 (d, J = 2.4 Hz, 1H), 8.03 (dd, J = 8.5, 2.4 Hz, 1H), 7.57 (d, J = 8.5 Hz, 1H), 3.92 (s, 2H), 3.78 (t, J = 6.5 Hz, 2H), 3.12 (t, J = 6.5 Hz, 2H), 1.43 (s, 9H).

[0176] Step 3: V1-3 (15.6 g, 49.41 mmol) was dissolved in dichloromethane (44.0 mL) and stirred at 20 °C, trifluoroacetic acid (28.17 g, 247.03 mmol) was added, and the reaction was stirred for 24 h. The reaction was concentrated under reduced pressure. The crude product was further dried by oil pump under reduced pressure, and purified by Combi Flash silica gel column (120 g, 0-38% ethyl acetate / petroleum ether) to give yellow oily liquid V1 (10.24 g, 79.83% yield). 1 H NMR (400 MHz, CDC13) δ 8.25 (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] + .

[0177] Preparation of intermediate compound V2

[0178] Step 1: V2-1 (413 mg, 1 mmol) was dissolved in N,N-dimethylformamide (10 mL), and tris(dibenzylideneacetone)dipalladium (47 mg, 0.05 mmol), 1,1'-bis(diphenylphosphino)ferrocene (28 mg, 0.05 mmol), zinc acetate (91 mg, 0.5 mmol), zinc cyanide (170 mg, 1.5 mmol) were added. The reaction was stirred at 120 °C for 2 h under nitrogen protection. The solvent was removed by concentration (i.e. concentrated dry), and the product was directly separated by silica gel column (dichloromethane:methanol = 100:1 to 9:1) to give brown solid V2-2 (390 mg, crude). LC-MS m / z (ESI): 288.0 [M+1] + .

[0179] Step 2: V2-2 (390 mg, 1.3 mmol) was dissolved in methanol (10 mL), and hydrochloric acid (1 mL), platinum dioxide (150 mg, 0.6 mmol) was added. The reaction was stirred under hydrogen for 15 h. The product was directly separated by silica gel column (dichloromethane:methanol = 100:1 to 2:1) to give off-white solid V2 (360 mg, 91.1% yield). LC-MS m / z (ESI): 292.0 [M+1] + .

[0180] Preparation of intermediate compound V3

[0181] 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 successively. The reaction was stirred at 20 °C for 4 h. The reaction solution was poured into a separatory funnel, water (300 mL) was added, and ethyl acetate (60 mL x 2) was used to extract the solution. The combined organic phase was washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting crude product was purified by Combi Flash silica gel column (120 g, 0-65% ethyl acetate / petroleum ether) to give yellow oily liquid V3-1 (10.41 g, 93.07% yield). LC-MS m / z (ESI): 273.1 [M+1] + .

[0182] Step 2: V3-1 (10.41 g, 38.18 mmol) was dissolved in tetrahydrofuran (60 mL) in a 500 mL three-necked flask, which was stirred in an ice-water bath at 0 °C under nitrogen protection. Borane tetrahydrofuran (1 M, 381.76 mmol, 382 mL) was added dropwise slowly. After the addition was completed, the reaction was heated and stirred in an oil bath at 70 °C for 12 h. After the reaction was cooled to room temperature, it was stirred in a water bath, and methanol was added dropwise slowly to quench the reaction until no gas bubbles were generated. The resulting liquid was dissolved in a small amount of methanol and stirred, and hydrochloric acid / dioxane (4 M, 191 mmol, 47.7 mL) was added. After stirring for 10 min, LC-MS detection showed that the intermediate was completely converted into the target product. The solution was concentrated under reduced pressure, dissolved in ethyl acetate (100 mL), and poured into a separatory funnel. Saturated sodium bicarbonate (200 mL) was added to adjust the pH of the aqueous solution to weak alkaline, and ethyl acetate (80 mL x 2) was used to extract the solution. The combined organic phase was washed with saturated brine (100 mL) and dried over anhydrous sodium sulfate. After filtration, the solution was concentrated under reduced pressure, and the resulting crude product was dried under reduced pressure by oil pump to give yellow oily liquid V3-2 (9.81 g, 79.46% yield). LC-MS m / z (ESI): 259.1 [M+1] + .

[0183] 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) and stirred at 20 °C, triethylamine (9.21 g, 91.01 mmol, 12.69 mL) was added slowly, after the addition was completed, the reaction was continued to stir for 12 hours. The reaction solution was rotary evaporated under reduced pressure. The obtained crude product was separated and purified by Combi Flash silica gel column (120 g, 0-16% ethyl acetate / petroleum ether) to obtain yellow oily liquid V3-3 (5.63 g, 51.72% yield). LC-MS m / z (ESI): 259.1 [M+1-100] + .

[0184] 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), iron powder (4.38 g, 78.45 mmol) was added, after the addition was completed, nitrogen was replaced, and the reaction was heated and stirred in an 80 °C oil bath for 4 hours. The cooled reaction solution was filtered through diatomite, the filter cake was washed with ethanol (80 mL x 3), the combined filtrate was rotary evaporated under reduced pressure. The obtained crude product was separated and purified by Combi Flash silica gel column (40 g, 0-28% ethyl acetate / petroleum ether) to obtain yellow oily liquid V3 (4.71 g, 91.29% yield). LC-MS m / z (ESI): 229.1 [M+1-100] + .

[0185] Preparation of intermediate compound V4

[0186] Step 1: V1 (2.59 g, 10 mmol) was dissolved in N,N-dimethylformamide (20 mL), trifluoroethylamine (1 g, 10 mmol), HATU (5.7 g, 1.5 mmol), triethylamine (2 g, 20 mmol) were added, and the reaction was carried out at room temperature for 1 hour, water was added, and ethyl acetate was extracted, and concentrated to dryness to obtain brown solid V4-1 (3.4 g, crude). LC-MS m / z (ESI): 341.0 [M+1] + .

[0187] Step 2: V4-1 (3.4 g, 10 mmol) was dissolved in tetrahydrofuran (15 mL), and borane tetrahydrofuran (60 mL) was added, and the reaction was carried out at 70 °C for 2 hours, and methanol was added to quench, and concentrated to dryness to obtain yellow oily substance V4-2 (3.9 g, crude). LC-MS m / z (ESI): 327.0 [M+1] + .

[0188] Step 3: V4-2 (3.9 g, 12 mmol) was dissolved in tetrahydrofuran (40 mL), di-tert-butyl dicarbonate (2.5 g, 12 mmol), triethylamine (2.5 g, 25 mmol) were added, and the reaction was stirred at room temperature for 12 hours. The reaction was concentrated to dryness and separated by silica gel column (ethyl acetate / petroleum ether = 31%) to give yellow oil V4-3 (2.9 g, 56.9% yield). LC-MS m / z (ESI): 327.0 [M+1-100] + .

[0189] Step 4: V4-3 (2.9 g, 6.8 mmol) was dissolved in ethanol (40 mL), water (20 mL), and iron powder (2.1 g, 40 mmol), ammonium chloride (2.1 g, 40 mmol) were added, and the reaction was stirred at 86 °C for 2 hours. The reaction was filtered, the filtrate was extracted, and concentrated to dryness to give yellow oil V4 (2 g, crude). LC-MS m / z (ESI): 297.0 [M+1-100] + .

[0190] Preparation of intermediate compound 3k

[0191] Step 1: Compound 3a (10 g, 56.13 mmol) was dissolved in anhydrous dichloromethane (80 mL), 2,2-dimethyl-1,3-dioxane-4,6-dione (7.69 g, 53.33 mmol) was added, and the reaction was reduced to 0 °C. 4-dimethylaminopyridine (3.43 g, 28.07 mmol), triethylamine (13.06 g, 129.11 mmol) were added, and finally solid EDCI (16.14 g, 84.20 mmol) was slowly added. The reaction was stirred at 0 °C for half an hour, then gradually increased to room temperature, and then reacted for 4 hours. LC-MS monitoring showed that the raw material was basically gone, and a new main peak was obtained as an intermediate state (LC-MS did not give the molecular weight). The reaction was concentrated to give brown oil crude 3b, which was directly used in the next step.

[0192] Step 2: The above dried crude 3b was added to toluene (250 mL) and concentrated hydrochloric acid (12 M, 35 mL), and the reaction was gradually heated to 65 °C with an oil bath for 12 hours. The reaction solution was carefully neutralized to pH about 9 with saturated aqueous sodium carbonate solution, then extracted with ethyl acetate three times, the combined organic phase was dried with sodium sulfate, washed with sodium bicarbonate aqueous solution, and concentrated to dryness under reduced pressure to give 3c (7.8 g, 78.8% yield) as a white solid. LC-MS m / z (ESI): 177.1 [M+1] + .

[0193] Step 3: Compound 3c (3.6 g, 20.43 mmol) was dissolved in anhydrous methanol (100 mL), sodium acetate (13.41 g, 163.48 mmol), hydroxylamine hydrochloride (11.36 g, 163.48 mmol) were added, the reaction was heated to 45 °C in an oil bath with stirring for 2 hours. The methanol was removed under reduced pressure, ethyl acetate and water were added, the organic phase was extracted twice with ethyl acetate, the combined organic phase was 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] + .

[0194] Step 4: Compound 3d (4.5 g, 23.54 mmol) was dissolved in anhydrous methanol (100 mL), wet palladium-carbon (1.09 g, 10% wt) was added, the reaction was heated to 40 °C after three times of hydrogen balloon replacement, and the reaction was stirred for 3 hours. After the reaction was cooled, diatomite was filtered, the filter cake was washed with methanol, and the product 3e (3.5 g, crude product) was used without drying, and the filtrate was directly used for the next reaction. LC-MS m / z (ESI): 178.1 [M+1] + .

[0195] Step 5: Compound 3e (3.5 g, 19.75 mmol) was dissolved in anhydrous methanol (120 mL) (this reaction system is the filtrate after the last step), triethylamine (6.38 g, 49.38 mmol, 8.60 mL), di-tert-butyl dicarbonate (6.47 g, 29.63 mmol, 6.81 mL) were added, and the reaction was stirred at room temperature for 1 hour. The reaction solution was dried under reduced pressure to give a brown oily crude product. After the brown oily substance was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 4: 1), compound 3f (3.6 g, 65.7% yield) was obtained as a brown oily substance; LC-MS m / z (ESI): 222.1 [M+1-56] + .

[0196] 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, and the reaction was stirred at room temperature for 1.5 hours. The reaction solution was neutralized to pH about 5 by 2M hydrochloric acid, and then the organic phase was extracted with ethyl acetate, and the combined organic phase was dried under reduced pressure to give compound 3g (3.5 g, 91.3% yield) as a white solid; LC-MS m / z (ESI): 317.8 [M+23] + .

[0197] Step 7: Compound 3g (1 g, 3.39 mmol) was dissolved in anhydrous methanol (6 mL) and ethyl acetate (6 mL), the reaction was cooled to -20 °C, then tri- methylsilanated diazomethane (2 M, 8.47 mL) was added slowly dropwise under argon protection, the reaction was stirred at -10 °C to room temperature for 1 h. The reaction was quenched carefully by adding water, then the organic phase was extracted with ethyl acetate, the combined organic phase was rotary evaporated under reduced pressure to give compound 3h (1.1 g, crude) as brown oil, which was used directly in the next step. LC-MS m / z (ESI): 332.2 [M+23] + .

[0198] Step 8: Compound 3h (0.85 g, 2.75 mmol) was dissolved in anhydrous tetrahydrofuran (15 mL), the reaction was cooled to 0 °C, then triphenylphosphine (1.08 g, 4.12 mmol), carbon tetrabromide (1.37 g, 4.12 mmol) was added, the reaction was stirred at room temperature for 2 h. The reaction was rotary evaporated under reduced pressure to give crude brown oil. The brown oil was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4: 1) to give compound 3i (166 mg, 16.2% yield) as brown oil; LC-MS m / z (ESI): 316.1, 318.1 [M+1-56] + .

[0199] Step 9: Compound 3i (160 mg, 429.82 μmol) was dissolved in anhydrous acetonitrile (7 mL), N, N-diisopropylethylamine (277.75 mg, 2.15 mmol) was added, the reaction was heated to 85 °C and stirred for 16 h. The reaction was rotary evaporated under reduced pressure to give crude brown oil, the brown oil was purified by silica gel column chromatography (dichloromethane: methanol = 10: 1) to give compound 3j (145 mg, 87.1% yield) as brown oil; LC-MS m / z (ESI): 388.2 [M+1] + .

[0200] Step 10: Compound 3j (160 mg, 412.98 μmol) was dissolved in anhydrous dichloromethane (3 mL), trifluoroacetic acid (706.34 mg, 6.19 mmol) was added, the reaction was stirred at room temperature for 1 h. The reaction was rotary evaporated under reduced pressure to give brown crude oil. The brown oil was purified by silica gel column chromatography (dichloromethane: methanol = 7: 1) to give compound 3k (105 mg, 88.5% yield) as brown oil; LC-MS m / z (ESI): 288.2 [M+1] + .

[0201] Preparation of intermediate compound 4i

[0202] Step 1 : Compound 3c (6 g, 34.06 mmol) and R-(+)-tert-butylsulfonamide (8.26 g, 68.12 mmol) were dissolved in dry tetrahydrofuran (90 mL), and tetraethyl orthotitanate (68.12 mmol, 14.28 mL) was added. After the reaction was protected with argon, it was heated to 75 °C and stirred for 20 hours. The dehydrated imine compound 4a was completely formed. The crude product was directly used in the next step reaction. LC-MS m / z (ESI): 280.1 [M+1] + .

[0203] Step 2: In another reaction flask, tetrahydrofuran (40 mL) and sodium borohydride (5.15 g, 136.23 mmol) were added, and the temperature was lowered to -60 °C. The tetrahydrofuran (90 mL) mixture containing the imine product 4a from the previous step was slowly added dropwise through a dropping funnel into the new reaction flask. After the addition was complete, the temperature was gradually increased to room temperature, and the reaction was allowed to proceed for 2 hours. After the reaction was complete, the reaction mixture was slowly added to an excess of aqueous solution, and a large amount of solid was precipitated. Bubbles were generated in the reaction, and care was taken to avoid material overflow. The reaction mixture was then filtered through diatomite, and the filter cake was washed with tetrahydrofuran until the filtrate was clear. The combined filtrate was extracted three times with ethyl acetate and twice with dichloromethane:methanol (10:1). The organic phase was dried over sodium sulfate and concentrated to give a brown solid. The brown solid was purified by column chromatography (dichloromethane:methanol = 20:1) to give compound 4b (4.8 g, 50.1% yield over two steps) as a brown solid. LC-MS m / z (ESI): 282.1 [M+1] + .

[0204] Step 3: Compound 4b (4.8 g, 17.06 mmol) was dissolved in dry dioxane (20 mL), and hydrochloric acid dioxane (4 M, 21.32 mL) was added. The reaction was stirred at room temperature for 0.5 hours. After the reaction was concentrated and dried, compound 4c (3.8 g, crude) was obtained as a brown solid. The product was directly used in the next step reaction. LC-MS m / z (ESI): 178.1 [M+1] + ;

[0205] Step 4: Compound 4c (3.5 g, 19.75 mmol) was dissolved in dry tetrahydrofuran (40 mL), water (40 mL), and sodium bicarbonate (6.64 g, 79.01 mmol) was added portionwise. BOC anhydride (8.62 g, 39.50 mmol) was added slowly, and the reaction was stirred at room temperature for 20 hours. Ethyl acetate was added, and the organic phase was combined, dried over sodium sulfate, and concentrated to give a crude oil. The crude product was purified by column chromatography on silica gel (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]+ .

[0206] Step 5 to Step 9: Refer to the preparation method of intermediate compound 3k, replace 3f with 4d to prepare intermediate 4i. LC-MS m / z (ESI): 288.1 [M+1] + .

[0207] Preparation of intermediate compound 12e

[0208] 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 h. The reaction was concentrated and the crude product was purified by silica gel column (petroleum ether: ethyl acetate = 20: 1) to give a colorless liquid which solidified to white solid 12b (2.6 g, 68.0% yield) after standing. 1 H NMR (400 MHz, CDC13) δ 7.73 ~ 7.68 (m, 2H), 4.88 (m, 3H), 3.95 (s, 2H).

[0209] Step 2: 12b (2.6 g, 7.98 mmol), 3-amino-2,6-piperidinedione hydrochloride (1.71 g, 10.37 mmol) and diisopropylethylamine (3.09 g, 23.93 mmol) were dissolved in acetonitrile (40 mL) and the reaction was stirred at 80 °C for 48 h. The reaction was concentrated and purified by silica gel column (dichloromethane: methanol = 10: 1) to directly give gray solid 12c (2.7 g, 99.23% yield). MS m / z (ESI): 341.0 [M+1] + .

[0210] Step 3: A solution of 12c (2.7 g, 7.91 mmol), zinc cyanide (1.71 g, 7.91 mmol), 1,1'-bis(diphenylphosphino)ferrocene (877 mg, 1.58 mmol) and tris(dibenzylideneacetone)dipalladium (724 mg, 0.791 mmol) in N,N-dimethylformamide (30 mL) was stirred at 120 °C for 12 h under nitrogen protection. The reaction was concentrated and purified by silica gel column (dichloromethane: methanol = 10: 1) to give gray solid 12d (2.0 g, 87.97% yield). LC-MS m / z (ESI): 288.0 [M+1] + .

[0211] Step 4: 12d (1 g, 3.48 mmol) was dissolved in methanol (15 mL), concentrated hydrochloric acid (1.5 mL) was added, and platinum dioxide (237.3 mg, 1.04 mmol) was added. The reaction was stirred at room temperature under a hydrogen atmosphere for 12 h. The reaction was filtered, the filter cake was washed with methanol (25 mL x 5), and the filtrate was combined and concentrated to give yellow solid 12e (1.1 g, 96.41% yield). MS m / z (ESI): 292.1 [M+1] + .

[0212] Preparation of intermediate compound 13d

[0213] Step 1: Compound V1 (2.59 g, 10 mmol) was dissolved in N,N-dimethylformamide (20 mL), ammonium chloride (2.1 g, 40 mmol), HATU (5.7 g, 1.5 mmol), triethylamine (2 g, 20 mmol) were added, and the reaction was stirred at room temperature for 1 h. Water was added, and the reaction was extracted with ethyl acetate, dried, and concentrated to give brown solid 13a (2.7 g, crude). LC-MS m / z (ESI): 259.0 [M+1] + .

[0214] Step 2: 13a (2.58 g, 10 mmol) was dissolved in tetrahydrofuran (15 mL), and borane tetrahydrofuran (60 mL) was added. The reaction was stirred at 70 °C for 2 h, dried, quenched with methanol, and dried to give yellow oil 13b (2.9 g, crude). LC-MS m / z (ESI): 245.0 [M+1] + .

[0215] Step 3: 13b (2.5 g, 10 mmol) was dissolved in tetrahydrofuran (40 mL), di-tert-butyl dicarbonate (2.5 g, 12 mmol), and triethylamine (2.5 g, 25 mmol) were added. The reaction was stirred at room temperature for 12 h, dried, and purified by column chromatography on silica gel (ethyl acetate / petroleum ether = 31 vol% / 69 vol%) to give yellow oil 13c (2.0 g, 56.8% yield). LC-MS m / z (ESI): 245.0 [M+1-100] + .

[0216] 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), ammonium chloride (0.6 g, 10 mmol) was added. The reaction was stirred at 86 °C for 2 h, filtered, the filtrate was extracted, and dried to give yellow oil 13d (0.5 g, crude). LC-MS m / z (ESI): 215.0 [M+1-100] + .

[0217] Preparation of intermediate compound 16c

[0218] Step 1 : 1 -Benzyl oxycarbonyl-3-pyrrolidinone (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) and stirred at 0 °C, then trimethylsilyl trifluoromethanesulfonate (1.29 g, 5.64 mmol) was added. After the addition was completed, the reaction was stirred for another 0.5 h, then concentrated to dryness to give yellow oil 16b (0.8 g, crude). LC-MS m / z (ESI): 405.0 [M+1] + .

[0219] Step 2: 16b (0.8 g, 2 mmol) was dissolved in ethanol (10 mL), water (5 mL), then reduced iron powder (0.6 g, 10 mmol), ammonium chloride (0.6 g, 10 mmol) were added, and the reaction was stirred at 86 °C for 2 h. The reaction mixture was filtered, the filtrate was extracted, concentrated to dryness, and purified by CombiFlash column chromatography (12 g, 0-36% ethyl acetate / petroleum ether) to give yellow oil 16c (0.5 g, 67.5% yield). LC-MS m / z (ESI): 375.0 [M+1] + .

[0220] Preparation of intermediate compound 22f

[0221] Step 1 : Compound 22a (491 mg, 2.02 mmol) and N-bromosuccinimide (431.37 mg, 2.42 mmol) were dissolved in dichloromethane (6 mL) and stirred at room temperature, then azobisisobutyronitrile (16.58 mg, 100.99 μmol) was added, and the reaction was stirred at 90 °C under nitrogen protection for 2 h. The reaction mixture was concentrated to dryness under reduced pressure, and the obtained crude product was purified by CombiFlash silica gel column chromatography to give compound 22b (570 mg, 87.64% yield). LC-MS m / z (ESI): 322.9 [M+1] + .

[0222] Step 2: Compound 22b (570 mg, 1.77 mmol) and 3-amino-2,6-piperidinedione (437.04 mg, 2.66 mmol, HC1) were dissolved in acetonitrile (5.0 mL), then diisopropylethylamine (686.35 mg, 5.31 mmol, 925.00 μί) was added, and the reaction was stirred at 80 °C under nitrogen protection for 24 h. After the reaction was cooled to room temperature, the reaction was concentrated under reduced pressure. The obtained crude product was purified by CombiFlash silica gel column to give compound 22c (171 mg, 28.65% yield). LC-MS m / z (ESI): 337.0 [M+1] + .

[0223] 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), then 1,1'-bis(diphenylphosphino)ferrocene (14.06 mg, 25.36 μmol) was added, followed by the addition of bis(dibenzylideneacetone)palladium (23.22 mg, 25.36 μmol) under nitrogen protection, and the reaction was stirred at 100 °C for 14 h. After the reaction was cooled to room temperature, the reaction was concentrated under reduced pressure. The obtained crude product was purified by CombiFlash silica gel column to give compound 22d (133 mg, 92.57% yield). LC-MS m / z (ESI): 284.0 [M+1] + .

[0224] 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) and stirred at 0 °C under nitrogen protection, then nickel dichloride hexahydrate (338.98 mg, 1.43 mmol) and sodium borohydride (40.47 mg, 1.07 mmol) were added successively. After the addition was completed, the reaction was stirred at room temperature for 16 h. The reaction was concentrated under reduced pressure, and the obtained crude product was purified by CombiFlash silica gel column to give compound 22e (84 mg, 60.81% yield). LC-MS m / z (ESI): 388.2 [M+1] + .

[0225] Step 5: Compound 22e (84 mg, 216.81 μmol) was dissolved in dichloromethane (1.5 mL) and stirred at 20 °C. Trifluoroacetic acid (247.21 mg, 2.17 mmol) was added. After the addition was completed, the reaction was stirred for another 12 h. The reaction was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (30 mL) and acetonitrile (10 mL), and basified by the addition of triethylamine (1.5 mL). The mixture was concentrated under reduced pressure again. The resulting crude product was purified by CombiFlash silica gel column to give compound 22f (30 mg, 48.16% yield). LC-MS m / z (ESI): 288.1 [M+1] + .

[0226] Example 1: Preparation of compound 1

[0227] Step 1: V3 (328 mg, 1 mmol) was dissolved in tetrahydrofuran (8 mL), and then triphosgene (120 mg, 0.4 mmol) was added. The reaction was stirred at room temperature for 0.5 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue was added to a solution of V2 (290 mg, 1 mmol) and triethylamine (600 mg, 6 mmol) in N,N-dimethylformamide (5 mL). The reaction was stirred at room temperature for 0.3 h. The reaction was concentrated under reduced pressure. The residue was separated by silica gel column (dichloromethane:methanol = 100:1 to 10:1) to give 1a (290 mg, 45.0% yield) as a yellow oil. LC-MS m / z (ESI): 546.0 [M+1-100] + .

[0228] Step 2: 1a (65 mg, 0.1 mmol) was dissolved in dichloromethane (5 mL), and then trifluoroacetic acid (2 mL) was added. The reaction was stirred at room temperature for 0.5 h. The reaction was directly concentrated to give a yellow oil. The yellow oil was directly separated by reverse phase HPLC (acid method, GILSON, Waters-SunFire C18 column; ACN / Water+0.04% FA; gradient, 10%-30%). After lyophilization, compound 1 (18 mg, 40.8% yield) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 10.91 (s, 1H), 9.00 (s, 1H), 7.63 (d, J = 2.0 Hz, 1H), 7.59 - 7.46 (m, 2H), 7.17 (dt, J = 8.4, 5.3 Hz, 2H), 7.03 (t, J = 5.9 Hz, 1H), 5.09 (dd, J = 13.3, 5.1 Hz, 1H), 4.55 (d, J = 17.4 Hz, 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, 16.5 Hz, 2H), 2.46 - 2.31 (m, 4H), 2.02 - 1.94 (m, 1H). LC-MS m / z (ESI): 546.2 [M+1] + .

[0229] Example 2: Preparation of compound 2

[0230] The title product compound 2 (130 mg, 65% yield) was prepared according to the synthetic procedure of Example 1 by replacing starting material V3 with V4. 1 H NMR (400 MHz, DMSO-d6) δ 10.91 (s, 1H), 9.00 (s, 1H), 7.63 (d, J = 2.0 Hz, 1H), 7.59 - 7.46 (m, 2H), 7.17 (dt, J = 8.4, 5.3 Hz, 2H), 7.03 (t, J = 5.9 Hz, 1H), 5.09 (dd, J = 13.3, 5.1 Hz, 1H), 4.55 (d, J = 17.4 Hz, 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, 16.5 Hz, 2H), 2.46 - 2.31 (m, 4H), 2.02 - 1.94 (m, 1H). LC-MS m / z (ESI): 546.2 [M+1] + .

[0231] Example 3: Preparation of compound 3

[0232] Step 1 : Compound intermediate V3 (90 mg, 273.69 μmol) was dissolved in anhydrous dichloromethane (5 mL), the reaction system was cooled to 0 °C, N,N- diisopropylethylamine (106.12 mg, 821.08 μmol, 143.01 μL) was added under argon protection, and then triphosgene (26.80 mg, 90.32 μmol) dissolved in dichloromethane (1 mL) was slowly added dropwise with a syringe. The reaction was kept in ice water bath for 1 h. Then compound 3k (78.64 mg, 273.69 μmol) dissolved in N,N-dimethylformamide (2 mL) was slowly added dropwise with a syringe. The reaction temperature was gradually increased to room temperature, and the reaction was carried out for 15 h. The reaction solution was concentrated by rotary evaporation under reduced pressure to obtain a brown oil. The brown oil was purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain compound 31 (125 mg, 71.1% yield) as a brown oil. LC-MS m / z (ESI): 541.8 [M+1-100] + .

[0233] Step 2: Compound 31 (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 was carried out at room temperature for 0.5 h. The reaction solution was concentrated by rotary evaporation under reduced pressure to obtain a brown oil. The brown oil was dried by rotary evaporation and then dried by oil pump. The product was separated by reverse phase HPLC preparation (acid method, GILSON, Waters-SunFire C18 column; ACN / Water+0.04% FA; gradient, 20%-50%) to obtain compound 3 (110 mg, 59.8% yield) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 11.02 (s, 1H), δ 9.12 (s, 1H), 8.38 (s, 1H), 7.69 (d, J = 7.8 Hz, 1H), 7.63 (d, J = 2.1 Hz, 1H), 7.55 (s, 1H), 7.48 (d, J = 7.9 Hz, 1H), 7.39 (s, 1H), 7.23 - 7.08 (m, 2H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.89 (p, J = 7.1 Hz, 1H), 4.45 (dd, J = 17.4, 2.9 Hz, 1H), 4.30 (dd, J = 17.4, 1.8 Hz, 1H), 3.55 (q, J = 6.4, 5.2 Hz, 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 = 7.0 Hz, 3H). LC-MS m / z (ESI): 542.2 [M+1] + .

[0234] Example 4: Preparation of compound 4

[0235] The title product compound 4 (57.2 mg, 55.6% yield) was prepared according to the preparation method of Example 3 by replacing starting material V3 with V4 and replacing starting material 3k with 4i. 1 H NMR (400 MHz, DMSO-d6) δ 11.00 (s, 1H), 8.56 (s, 1H), 7.70 (d, J = 7.9 Hz, 1H), 7.70 (d, J = 2.1 Hz, 1H), 7.55 (s, 1H), 7.48 (d, J = 7.9 Hz, 1H), 7.19 (d, J = 8.4 Hz, 1H), 7.09 (dd, J = 8.4, 2.1 Hz, 1H), 6.84 (d, J = 7.4 Hz, 1H), 5.11 (dd, J = 13.3, 5.2 Hz, 1H), 4.90 (t, J = 7.1 Hz, 1H), 4.46 (d, J = 17.0 Hz, 1H), 4.31 (d, J = 17.3 Hz, 1H), 3.54 (t, J = 7.0 Hz, 2H), 3.46 (t, J = 5.6 Hz, 2H), 3.21 (q, J = 10.3 Hz, 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 = 6.9 Hz, 3H). LC-MS m / z (ESI): 610.1 [M+1] + .

[0236] Example 5: Preparation of compound 5

[0237] The title product compound 5 (48.3 mg, 32% yield) was prepared according to the preparation method of Example 4 by replacing the starting material R-(+)-tert-butylsulfonamide with S-(-)-tert-butylsulfonamide. 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.56 (s, 1H), 7.70 (d, J = 7.9 Hz, 1H), 7.62 (d, J = 2.1 Hz, 1H), 7.55 (s, 1H), 7.48 (d, J = 7.9 Hz, 1H), 7.19 (d, J = 8.4 Hz, 1H), 7.09 (dd, J = 8.4, 2.1 Hz, 1H), 6.83 (d, J = 7.4 Hz, 1H), 5.10 (dd, J = 13.3, 5.2 Hz, 1H), 4.90 (t, J = 7.1 Hz, 1H), 4.45 (d, J = 17.0 Hz, 1H), 4.31 (d, J = 17.3 Hz, 1H), 3.53 (t, J = 7.0 Hz, 2H), 3.43 (t, J = 5.6 Hz, 2H), 3.21 (q, J = 10.3 Hz, 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 = 6.9 Hz, 3H). LC-MS m / z (ESI): 610.1 [M+1] + .

[0238] Example 6: Preparation of compound 6

[0239] The title product compound 6 (71.93 mg, 17.39% yield) was prepared according to the preparation method of Example 1 by replacing the starting material methylamine with 2,2-difluoroethylamine. 1H NMR (400 MHz, DMSO-d6) δ 11.01 (s, 1H), 8.97 (s, 1H), 7.65 (d, J = 2.0 Hz, 1H), 7.57 (d, J = 7.7 Hz, 1H), 7.51 (t, J = 7.0 Hz, 1H), 7.24 - 7.11 (m, 2H), 7.00 (t, J = 6.0 Hz, 1H), 6.09 (tt, J = 55.6, 4.1 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.59 - 4.28 (m, 4H), 3.61 - 3.47 (m, 4H), 3.08 (td, J = 15.8, 4.1 Hz, 2H), 2.95 - 2.77 (m, 6H), 2.59 (dd, J = 15.4, 2.7 Hz, 1H), 2.41 (td, J = 13.2, 4.5 Hz, 1H), 2.05 - 1.94 (m, 1H). LC-MS m / z (ESI): 596.2 [M+1] + .

[0240] Example 7: Preparation of compound 7

[0241] The title product compound 7 (4.8 mg, 15.0% yield) was prepared according to the preparation method of Example 1 by replacing the raw material methylamine with cyclopropylamine. 1 H NMR (400 MHz, DMSO-d6) δ 11.01 (s, 1H), 8.97 (s, 1H), 7.65 (d, J = 2.0 Hz, 1H), 7.57 (d, J = 7.7 Hz, 1H), 7.51 (t, J = 7.0 Hz, 1H), 7.24 - 7.11 (m, 2H), 7.00 (t, J = 6.0 Hz, 1H), 6.09 (tt, J = 55.6, 4.1 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.59 - 4.28 (m, 4H), 3.61 - 3.47 (m, 4H), 3.08 (td, J = 15.8, 4.1 Hz, 2H), 2.95 - 2.77 (m, 6H), 2.59 (dd, J = 15.4, 2.7 Hz, 1H), 2.41 (td, J = 13.2, 4.5 Hz, 1H), 2.05 - 1.94 (m, 1H). LC-MS m / z (ESI): 596.2 [M+1] + .

[0242] Example 8: Preparation of compound 8

[0243] The title product compound 8 (5.1 mg, 15.0% yield) was prepared according to the preparation method of Example 1 by replacing the starting material methylamine with (1R,2S)-2-fluorocyclopropane-1 -amine. 1 H NMR (400 MHz, DMSO-d6) δ 11.00 (s, 1H), 8.86 (s, 1H), 7.76 - 7.68 (m, 1H), 7.64 (s, 1H), 7.57 (d, J = 7.6 Hz, 1H), 7.51 (d, J = 5.9 Hz, 1H), 7.19 (d, J = 13.5 Hz, 1H), 6.87 (s, 1H), 5.17 - 5.02 (m, 1H), 4.53 (s, 1H), 4.40 (d, J = 8.4 Hz, 2H), 3.76 (d, J = 34.9 Hz, 1H), 3.62 - 3.57 (m, 2H), 3.54 - 3.41 (m, 2H), 3.07 (s, 2H), 2.96 (d, J = 4.8 Hz, 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] + .

[0244] Example 9: Preparation of compound 9

[0245] The title product compound 9 (17.32 mg, 46.39% yield) was prepared according to the preparation method of Example 1 by replacing the starting material methylamine with 2-(methylsulfonyl)ethylamine. 1H NMR (400 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.83 (s, 1H), 8.12 (s, 1H), 7.63 (d, J = 2.0 Hz, 1H), 7.57 (d, J = 7.7 Hz, 1H), 7.51 (t, J = 7.0 Hz, 1H), 7.21 (d, J = 8.4 Hz, 1H), 7.15 (d, J = 8.7 Hz, 1H), 6.83 (s, 1H), 5.09 (dd, J = 13.5, 5.1 Hz, 1H), 4.55 (d, J = 17.3 Hz, 1H), 4.42 (d, J = 5.8 Hz, 2H), 4.37 (d, J = 17.3 Hz, 1H), 3.57 (d, J = 7.0 Hz, 4H), 3.24 - 3.17 (m, 3H), 3.06 (s, 3H), 3.00 (s, 2H), 2.87 (d, J = 7.2 Hz, 3H), 2.59 (d, J = 17.2 Hz, 1H), 2.04 - 1.94 (m, 2H), 1.04 (t, J = 7.0 Hz, 1H). LC-MS m / z (ESI): 638.2 [M+1] + .

[0246] Example 10: Preparation of compound 10

[0247] The title product compound 10 (50 mg, 37% yield) was prepared according to the preparation method of Example 1 by replacing the starting material methylamine with 2-aminoacetonitrile. 1 H NMR (400 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.83 (s, 1H), 8.12 (s, 1H), 7.63 (d, J = 2.0 Hz, 1H), 7.57 (d, J = 7.7 Hz, 1H), 7.51 (t, J = 7.0 Hz, 1H), 7.21 (d, J = 8.4 Hz, 1H), 7.15 (d, J = 8.7 Hz, 1H), 6.83 (s, 1H), 5.09 (dd, J = 13.5, 5.1 Hz, 1H), 4.55 (d, J = 17.3 Hz, 1H), 4.42 (d, J = 5.8 Hz, 2H), 4.37 (d, J = 17.3 Hz, 1H), 3.57 (d, J = 7.0 Hz, 4H), 3.24 - 3.17 (m, 3H), 3.06 (s, 3H), 3.00 (s, 2H), 2.87 (d, J = 7.2 Hz, 3H), 2.59 (d, J = 17.2 Hz, 1H), 2.04 - 1.94 (m, 2H), 1.04 (t, J = 7.0 Hz, 1H). LC-MS m / z (ESI): 638.2 [M+1] + .

[0248] Example 11: Preparation of compound 11

[0249] Referring to the preparation method of Example 1, the raw material methylamine was replaced with 3,3-difluorocyclobutylamine to prepare the title product compound 11 (130 mg, 65% yield). 1 H NMR (400 MHz, DMSO-d6) δ 11.01 (s, 1H), 8.85 (s, 1H), 8.12 (s, 1H), 7.63 (s, 1H), 7.57 (d, J = 7.9 Hz, 1H), 7.51 (d, J = 6.8 Hz, 1H), 7.22 (d, J = 8.6 Hz, 1H), 7.16 (d, J = 10.5 Hz, 1H), 6.85 (s, 1H), 5.14-5.08 (m, 1H), 4.55 (d, J = 17.5 Hz, 1H), 4.40 (dd, J = 20.2, 11.7 Hz, 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 = 14.1 Hz, 2H), 2.58 (d, J = 14.8 Hz, 1H), 2.41 (d, J = 12.2 Hz, 3H), 1.98 (s, 1H). LC-MS m / z (ESI): 622.2 [M+1] + .

[0250] Example 12: Preparation of compound 12

[0251] Step 1: To a solution of compound V3 (60 mg, 182.46 μmol), diisopropylethylamine (47.16 mg, 364.93 μmol, 63.56 μL) in N,N-dimethylformamide (2 mL) was added N,N'-carbonyldiimidazole (29.59 mg, 182.46 μmol) in portions at 0 °C, slowly warmed to room temperature, continue to stir for 1 hour, then the above reaction solution was added dropwise to diisopropylethylamine (47.16 mg, 364.93 μmol, 63.56 μL) and 12e (53.15 mg, 182.46 μmol) in N,N-dimethylformamide (2 mL), the reaction solution was stirred for 1 hour. The reaction solution was concentrated and purified by silica gel column (dichloromethane:methanol = 20:1) to obtain yellow solid 12f (35 mg, 29.69% yield). MS m / z (ESI): 546.2 [M+1-100] + .

[0252] Step 2: A solution of 12f (35 mg, 54.17 μmol) in trifluoroacetic acid (0.2 mL) and dichloromethane (1 mL) was prepared at room temperature and stirred for 1 h. The reaction was concentrated and purified by preparative high performance liquid chromatography (acidic method, GILSON, Waters-SunFire C18; ACN / Water+0.04% FA; gradient, 10%-65%) to give compound 12 as a white solid (8.69 mg, 22.52% yield). 1 H NMR (400 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.90 (s, 1H), 8.39 (br s, 2H), 7.65 (d, J = 4.0 Hz, 1H), 7.58 (d, J = 4.0 Hz, 1H), 7.53 (d, J = 12.0 Hz, 1H), 7.23 (d, J = 4.0 Hz, 1H), 7.17 (dd, J = 8.0, 4.0 Hz, 1H), 6.89 (t, J = 4.0 Hz, 1H), 5.10 (dd, J = 12.0, 2.0 Hz, 1H), 4.46 - 4.28 (m, 4H), 3.63 (t, J = 8.0 Hz, 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] + .

[0253] Example 13: Preparation of compound 13

[0254] Step 1: After 13d (314 mg, 1 mmol) was dissolved in tetrahydrofuran (8 mL), triphosgene (120 mg, 0.4 mmol) was added and the reaction was stirred at room temperature for 0.5 h. After being dried, it was added to a solution of compound V2 (290 mg, 1 mmol) and triethylamine (600 mg, 6 mmol) in N,N-dimethylformamide (5 mL) and the reaction was stirred at room temperature for 0.3 h. After being dried, the yellow oil 13e (350 mg, 55.5% yield) was obtained directly by silica gel column chromatography (dichloromethane:methanol = 100:1 to 10:1). LC-MS m / z (ESI): 532.0 [M+1-100] + .

[0255] Step 2: After 13e (230 mg, 0.38 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (2 mL) was added and the reaction was allowed to proceed at room temperature for 0.5 h. The reaction was directly concentrated by rotary evaporation to give yellow oil 13f (390 mg, crude). LC-MS m / z (ESI): 532.0 [M+1] + .

[0256] Step 3: After 13f (390 mg, 0.7 mmol) was dissolved in dichloromethane (3 mL), acetic acid (3 mL), 3-oxetanone (400 mg, 5.6 mmol) was added and the reaction was allowed to proceed at 60 °C for 2 h. Sodium borohydride (400 mg, 2.3 mmol) was added and the reaction was allowed to proceed at 60 °C for 2 h. The reaction was concentrated and purified by silica gel column (dichloromethane: methanol = 100:1 to 8:1) to give yellow oil. The product was further purified by preparative HPLC (acidic method, GILSON, Waters-SunFire C18; ACN / Water+0.04% FA; gradient, 5%-35% ACN). After lyophilization, white solid compound 13 (60 mg, 15% yield) was obtained. 1 H NMR (400 MHz, DMSO-d6) d 11.01 (s, 1H), 8.85 (s, 1H), 8.43 (s, 1H), 7.63 (s, 1H), 7.57 (d, J = 7.9 Hz, 1H), 7.51 (d, J = 6.8 Hz, 1H), 7.22 (d, J = 8.6 Hz, 1H), 7.16 (d, J = 10.5 Hz, 1H), 6.85 (s, 1H), 5.14-5.08 (m, 1H), 4.55 (d, J = 17.5 Hz, 1H), 4.40 (dd, J = 20.2, 11.7 Hz, 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 = 14.1 Hz, 2H), 2.58 (d, J = 14.8 Hz, 1H), 2.41 (d, J = 12.2 Hz, 3H), 1.98 (s, 1H). LC-MS m / z (ESI): 588.2 [M+1] + .

[0257] Example 14: Preparation of compound 14

[0258] The title product compound 14 (15 mg, 14% yield) was prepared according to the procedure described in Example 13, by replacing 3-oxetanone with cyclobutanone. 1H NMR (400 MHz, DMSO-d6) δ 11.00 (s, 1H), 8.89 (s, 1H), 7.64 (s, 1H), 7.56 (d, J = 7.7 Hz, 1H), 7.50 (s, 1H), 7.19 (s, 1H), 7.15 (s, 1H), 6.90 (s, 1H), 5.10 (d, J = 8.4 Hz, 1H), 4.55 (d, J = 17.0 Hz, 1H), 4.39 (dd, J = 19.4, 11.2 Hz, 3H), 3.51 (s, 4H), 3.13 (s, 4H), 2.82 (s, 3H), 2.63 (d, J = 20.0 Hz, 2H), 2.43 - 2.39 (m, 3H), 1.58 (s, 3H). LC-MS m / z (ESI): 586.2 [M+1]

[0259] 1H), 2.00 (s, 3H), 1.58 (s, 3H). LC-MS m / z (ESI): 586.2 [M+1] + .

[0260] Example 15: Preparation of compound 15

[0261] The title product compound 15 (15 mg, 14% yield) was prepared according to the preparation method of Example 13 by replacing 3-oxetanone with (3,3-difluorocyclobutyl) methanol. 1 H NMR (400 MHz, DMSO-d6) δ 11.00 (s, 1H), 8.89 (s, 1H), 7.64 (s, 1H), 7.56 (d, J = 7.7 Hz, 1H), 7.50 (s, 1H), 7.19 (s, 1H), 7.15 (s, 1H), 6.90 (s, 1H), 5.10 (d, J = 8.4 Hz, 1H), 4.55 (d, J = 17.0 Hz, 1H), 4.39 (dd, J = 19.4, 11.2 Hz, 3H), 3.51 (s, 4H), 3.13 (s, 4H), 2.82 (s, 3H), 2.63 (d, J = 20.0 Hz, 2H), 2.43 - 2.39 (m, + .

[0262] Example 16: Preparation of compound 16

[0263] Step 1: 16c (188 mg, 0.5 mmol) was dissolved in tetrahydrofuran (6 mL), then triphosgene (60 mg, 0.2 mmol) was added, and the reaction was allowed to proceed at room temperature for 0.5 h. After being dried by rotary evaporation, the residue was added to a solution of V2 (145 mg, 0.5 mmol) and triethylamine (300 mg, 3 mmol) in N,N-dimethylformamide (5 mL), and the reaction was allowed to proceed at room temperature for 0.3 h. After being dried by rotary evaporation, the residue was separated by column chromatography (dichloromethane:methanol = 100:1 to 10:1) to directly obtain yellow oil 16d (180 mg, 51.8% yield). LC-MS m / z (ESI): 692.0 [M+1] + .

[0264] Step 2: 16d (180 mg, 0.26 mmol) was dissolved in methanol (5 mL), then palladium-carbon (40 mg) was added, and the reaction was allowed to proceed at room temperature under hydrogen for 0.5 h. After the reaction was completed, the reaction mixture was directly filtered and dried by rotary evaporation to obtain yellow oil. The yellow oil was directly subjected to preparative high-performance liquid chromatography separation (acid method, GILSON, Waters-SunFire C18; ACN / water+0.04% FA; gradient, 5%-30% ACN), and white solid compound 16 (30 mg, 20.8% yield) was obtained after freeze-drying. 1 H NMR (400 MHz, DMSO-d6) δ 9.42 (s, 1H), 8.40 (s, 1H), 7.65 (s, 1H), 7.56 (d, J = 7.8 Hz, 1H), 7.53-7.45 (m, 2H), 7.19 (s, 2H), 5.09 (d, J = 13.1 Hz, 1H), 4.55 (d, J = 17.4 Hz, 1H), 4.45-4.29 (m, 4H), 3.77 (s, 2H), 3.53 (dd, J = 21.4, 6.2 Hz, 7H), 2.91-2.82 (m, 4H), 2.58 (d, J = 17.2 Hz, 1H), 1.99 (s, 1H). LC-MS m / z (ESI): 558.2 [M+1] + .

[0265] Example 17: Preparation of compound 17

[0266] The title product compound 17 (43 mg, 33.8% yield) was prepared according to the preparation method of Example 16, by replacing 1-benzyloxycarbonyl-3-pyrrolidinone with N-benzyloxycarbonyl-2-pyrrolidinecarboxaldehyde. 1H NMR (400 MHz, DMSO-d6) δ 11.11 - 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, 5.0 Hz, 1H), 4.55 (d, J = 17.2 Hz, 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 = 6.9 Hz, 3H), 2.84 (t, J = 6.9 Hz, 2H), 2.58 (d, J = 15.0 Hz, 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] + .

[0267] Example 18: Preparation of compound 18

[0268] The title product compound 18 (25 mg, 16.8% yield) was prepared according to the preparation method of Example 16 by replacing the starting material 1-benzyloxycarbonyl-3-pyrrolidinone with 3-formylazetidine-1-carboxylic acid benzyl ester. 1 H NMR (400 MHz, DMSO-d6) δ 9.42 (s, 1H), 8.40 (s, 1H), 7.65 (s, 1H), 7.56 (d, J = 7.8 Hz, 1H), 7.53 - 7.45 (m, 2H), 7.19 (s, 2H), 5.09 (d, J = 13.1 Hz, 1H), 4.55 (d, J = 17.4 Hz, 1H), 4.45 - 4.29 (m, 4H), 3.77 (s, 2H), 3.53 (dd, J = 21.4, 6.2 Hz, 7H), 2.91 - 2.82 (m, 4H), 2.58 (d, J = 17.2 Hz, 1H), 1.99 (s, 1H). LC-MS m / z (ESI): 558.2 [M+1] + .

[0269] Example 19: Preparation of compound 19

[0270] The title product compound 19 (4.15 mg, 8.68% yield) was prepared according to the preparation method of Example 16 by replacing the starting material 1-benzyloxycarbonyl-3-pyrrolidinone with N-Cbz-piperidin-3-one. 1H NMR (400 MHz, DMSO-d6) δ 11.01 (s, 1H), 8.92 (s, 1H), 8.26 (s, 1H), 7.63 (d, J = 2.1 Hz, 1H), 7.57 (d, J = 7.7 Hz, 1H), 7.53 - 7.48 (m, 1H), 7.21 (d, J = 8.4 Hz, 1H), 7.15 (dd, J = 8.6, 2.2 Hz, 1H), 6.93 (t, J = 6.0 Hz, 1H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.55 (d, J = 17.4 Hz, 1H), 4.42 (d, J = 5.8 Hz, 2H), 4.37 (d, J = 17.4 Hz, 1H), 3.57 (t, J = 7.2 Hz, 2H), 3.48 - 3.40 (m, 2H), 3.04 (d, J = 13.6 Hz, 1H), 2.82 (t, J = 7.0 Hz, 2H), 2.75 - 2.66 (m, 2H), 2.63 - 2.54 (m, 2H), 1.98 (d, J = 13.6 Hz, 1H), 1.77 (s, 1H), 1.68 (s, 1H), 1.44 (s, 3H). LC-MS m / z (ESI): 571.8 [M+1] + .

[0271] Example 20: Preparation of Compound 20

[0272] The title product compound 20 (4.54 mg, 3.84% yield) was prepared according to the preparation method of Example 16 by replacing 1-benzyloxycarbonyl-3-pyrrolidinone with 1-benzyloxycarbonyl-3-azetidinone. 1 H NMR (400 MHz, DMSO-d6) δ 11.01 (s, 1H), 8.92 (s, 1H), 8.26 (s, 1H), 7.63 (d, J = 2.1 Hz, 1H), 7.57 (d, J = 7.7 Hz, 1H), 7.53 - 7.48 (m, 1H), 7.21 (d, J = 8.4 Hz, 1H), 7.15 (dd, J = 8.6, 2.2 Hz, 1H), 6.93 (t, J = 6.0 Hz, 1H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.55 (d, J = 17.4 Hz, 1H), 4.42 (d, J = 5.8 Hz, 2H), 4.37 (d, J = 17.4 Hz, 1H), 3.57 (t, J = 7.2 Hz, 2H), 3.48 - 3.40 (m, 2H), 3.04 (d, J = 13.6 Hz, 1H), 2.82 (t, J = 7.0 Hz, 2H), 2.75 - 2.66 (m, 2H), 2.63 - 2.54 (m, 2H), 1.98 (d, J = 13.6 Hz, 1H), 1.77 (s, 1H), 1.68 (s, 1H), 1.44 (s, 3H). LC-MS m / z (ESI): 571.8 [M+1] + .

[0273] Example 21: Preparation of compound 21

[0274] Step 1: After 13f (531 mg, 1 mmol) was dissolved in dichloromethane (10 mL) and methanol (3 mL), 2-oxoacetic acid (740 mg, 10 mmol), Ti(OiPr)4(0.8 mL) were added, and the reaction was carried out at 60 °C for 2 h, sodium borohydride (750 mg, 3 mmol) was added, 2-oxoacetic acid (740 mg, 10 mmol) was added, and the reaction was carried out at 60 °C for 4 h, H2O (5 mL) was added, and the mixture was filtered, washed with dichloromethane / methanol, and concentrated, and the product (210 mg) was obtained as an oil directly by silica gel column chromatography (dichloromethane:methanol = 100:1 to 3:1). The title compound 21 (63 mg, 11% yield) was obtained by preparative high performance liquid chromatography separation (acid method, GILSON, Waters-SunFire C18; ACN / Water+0.04% FA; gradient, 5%-25% ACN) and freeze-drying. 1 H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 10.98 (s, 1H), 7.64 (d, J = 2.1 Hz, 1H), 7.57 (d, J = 7.7 Hz, 1H), 7.53-7.47 (m, 1H), 7.19 (d, J = 8.4 Hz, 1H), 7.12 (dd, J = 8.4, 2.1 Hz, 1H), 6.80 (t, J = 5.8 Hz, 1H), 5.10 (dd, J = 13.3, 5.3 Hz, 1H), 4.55 (d, J = 17.2 Hz, 1H), 4.40 (dd, J = 19.9, 11.5 Hz, 3H), 3.52 (t, J = 7.0 Hz, 2H), 3.42 (t, J = 5.6 Hz, 2H), 3.28-3.11 (m, 2H), 2.91 (d, J = 13.9 Hz, 1H), 2.82 (t, J = 7.0 Hz, 2H), 2.71 (s, 2H), 2.66-2.54 (m, 2H), 2.36 (d, J = 38.3 Hz, 1H), 2.19 (s, 1H), 1.97 (s, 1H). LC-MS m / z (ESI): 544.1 [M+1] + .

[0275] Example 22: Preparation of compound 22

[0276] Referring to the synthetic method of Example 1, the starting material V2 was replaced by compound 22f to prepare the title product compound 22 (19.26 mg, 58.95% yield). 1H NMR (400 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.92 (s, 1H), 8.30 (s, 1H), 7.65 (d, J = 2.1 Hz, 1H), 7.53 (d, J = 7.8 Hz, 1H), 7.40 (d, J = 7.9 Hz, 1H), 7.24 - 7.12 (m, 2H), 6.93 (t, J = 5.6 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.42 (d, J = 17.1 Hz, 1H), 4.37 (d, J = 5.7 Hz, 2H), 4.25 (d, J = 17.1 Hz, 1H), 3.56 (q, J = 5.7, 4.4 Hz, 4H), 2.88 (dt, J = 19.6, 6.1 Hz, 4H), 2.63 - 2.56 (m, 1H), 2.43 (s, 3H), 2.38 (dd, J = 13.3, 8.8 Hz, 2H), 2.27 (s, 3H), 2.03 - 1.94 (m, 1H). LC-MS m / z (ESI): 541.8 [M+1] + .

[0277] Example 23: Preparation of compounds 23-1 and 23-2

[0278] The title product compound 23-1 (26.52 mg, 22.74% yield) was prepared according to the preparation method of Example 1 by replacing the raw material methylamine with (S)-1,1,1-trifluoropropan-2-amine. 1 H NMR (400 MHz, DMSO-d6) δ 11.01 (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), 7.52 ~ 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, 8.0 Hz, 1H), 4.56 (d, J = 20.0 Hz, 1H), 4.44 ~ 4.37 (m, 3H), 3.56 ~ 3.52 (m, 2H), 3.44 ~ 3.40 (m, 2H), 3.26 (br s, 1H), 2.96 ~ 2.82 (m, 3H), 2.73 (br s, 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] + .

[0279] The title product compound 23-2 (22.52 mg, 22.72% yield) was prepared according to the procedure of Example 1 by replacing the starting material methylamine with (R)-1,1,1-trifluoropropan-2-amine. 1 H NMR (400 MHz, DMSO-d6) δ 11.01 (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, 4.0 Hz, 1H), 4.56 (d, J = 20.0 Hz, 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), 2.76-2.70 (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] + .

[0280] Example 24: Preparation of compound 24

[0281] The title product compound 24 (130 mg, 65% yield) was prepared according to the synthetic procedure of Example 1 by replacing the starting material V2 with 12e and replacing the starting material V3 with V4. 1H NMR (400 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.87 (s, 1H), 8.32 (s, 1H), 7.65 (d, J = 2.1 Hz, 1H), 7.56 (d, J = 6.3 Hz, 1H), 7.50 (d, J = 8.8 Hz, 1H), 7.19 (d, J = 8.4 Hz, 1H), 7.12 (dd, J = 8.2, 2.2 Hz, 1H), 6.85 (t, J = 6.0 Hz, 1H), 5.09 (dd, J = 13.3, 5.0 Hz, 1H), 4.46 - 4.41 (m, 1H), 4.40 (s, 2H), 4.28 (d, J = 17.3 Hz, 1H), 3.52 (t, J = 7.0 Hz, 2H), 3.42 (t, J = 5.8 Hz, 2H), 3.20 (q, J = 10.6 Hz, 2H), 2.82 (t, J = 7.1 Hz, 2H), 2.70 (t, J = 5.6 Hz, 2H), 2.67 - 2.63 (m, 1H), 2.62 - 2.58 (m, 1H), 2.35 (d, J = 13.2 Hz, 1H), 1.98 (d, J = 9.7 Hz, 1H). LC-MS m / z (ESI): 614.2 [M+1] + .

[0282] Example 25: Preparation of compound 25

[0283] Step 1: Compound 21 (20 mg, 0.03 mmol) was dissolved in DMF (2 mL), added ammonium chloride (28 mg, 0.5 mmol), HATU (18 mg, 0.045 mmol), triethylamine (11 mg, 0.1 mmol), reacted at room temperature for 1 hour, concentrated dry, directly sent to preparative high performance liquid chromatography separation (acid method, GILSON, Waters-SunFire C18; ACN / Water+0.04%FA; gradient, 10%-30%ACN), after freeze-drying to obtain compound 25 (6 mg, 30.1% yield). 1H NMR (400 MHz, DMSO-d6) δ 10.99 (s, 1H), 7.64 (d, J = 2.1 Hz, 1H), 7.57 (d, J = 7.7 Hz, 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, 2.1 Hz, 1H), 6.80 (t, J = 5.8 Hz, 1H), 5.10 (dd, J = 13.3, 5.3 Hz, 1H), 4.55 (d, J = 17.2 Hz, 1H), 4.40 (dd, J = 19.9, 11.5 Hz, 3H), 3.52 (t, J = 7.0 Hz, 2H), 3.42 (t, J = 5.6 Hz, 2H), 3.28 - 3.11 (m, 2H), 2.91 (d, J = 13.9 Hz, 1H), 2.82 (t, J = 7.0 Hz, 2H), 2.71 (s, 2H), 2.66 - 2.54 (m, 2H), 2.36 (d, J = 38.3 Hz, 1H), 2.19 (s, 1H), 1.98 (s, 1H). LC-MS m / z (ESI): 589.2 [M+1] + .

[0284] Example 26: Preparation of compound 26

[0285] The title product compound 26 (23.43 mg, 29.46% yield) was prepared by following the synthetic procedure of example 1, replacing the starting material methylamine with 2-aminoethanesulfonamide. 1 H NMR (400 MHz, DMSO-d6) δ 11.01 (s, 1H), 8.84 (s, 1H), 8.14 (s, 1H), 7.66 (s, 1H), 7.65 ~ 7.51 (m, 2H), 7.22 (d, J = 8.0 Hz, 1H), 7.15 (d, J = 8.0 Hz, 1H), 6.91 ~ 6.83 (m, 3H), 5.11 (dd, J = 16.0, 8.0 Hz, 1H), 4.56 (d, J = 20.0 Hz, 1H), 4.48 ~ 4.31 (m, 3H), 3.58 ~ 3.49 (m, 4H), 3.20 ~ 3.16 (m, 2H), 3.03 ~ 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] + .

[0286] Example 27: Preparation of compound 27

[0287] The title product compound 27 (45.3 mg, 32% yield) was prepared according to the procedure of Example 4 by replacing the raw material R-(+)-tert-butylsulfonamide with S-(-)-tert-butylsulfonamide and replacing the raw material V4 with V3. 1 H NMR (400 MHz, DMSO-d6) δ 9.04 (s, 1H), 8.34 (s, 1H), 7.69 (d, J = 7.9 Hz, 1H), 7.63 (d, J = 2.0 Hz, 1H), 7.55 (s, 1H), 7.48 (d, J = 7.9 Hz, 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 = 7.1 Hz, 1H), 4.45 (dd, J = 17.4, 3.0 Hz, 1H), 4.31 (d, J = 17.3 Hz, 1H), 3.55 (q, J = 6.6, 5.9 Hz, 6H), 3.01 - 2.54 (m, 6H), 2.40 (s, 3H), 1.99 (s, 1H), 1.40 (d, J = 7.0 Hz, 3H). LC-MS m / z (ESI): 542.2 [M+1] + .

[0288] Example 28: Preparation of compound 28

[0289] The title product compound 28 (187 mg, 33.32% yield) was prepared according to the procedure of Example 1 by replacing the raw material methylamine with ethylamine. 1 H NMR (400 MHz, DMSO-d6) δ 9.04 (s, 1H), 8.34 (s, 1H), 7.69 (d, J = 7.9 Hz, 1H), 7.63 (d, J = 2.0 Hz, 1H), 7.55 (s, 1H), 7.48 (d, J = 7.9 Hz, 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 = 7.1 Hz, 1H), 4.45 (dd, J = 17.4, 3.0 Hz, 1H), 4.31 (d, J = 17.3 Hz, 1H), 3.55 (q, J = 6.6, 5.9 Hz, 6H), 3.01 - 2.54 (m, 6H), 2.40 (s, 3H), 1.99 (s, 1H), 1.40 (d, J = 7.0 Hz, 3H). LC-MS m / z (ESI): 542.2 [M+1] + .

[0290] Example 29: Preparation of compound 29

[0291] Reference to the preparation method of Example 1, the raw material methylamine is replaced by ethanolamine to prepare the title product compound 29 (340 mg, 29.63% yield). 1 H NMR (400 MHz, DMSO-d6) δ 11.03 (s, 1H), 9.23 (d, J = 11.4 Hz, 1H), 8.29 (s, 1H), 7.71 - 7.46 (m, 3H), 7.35 - 7.13 (m, 3H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.57 (d, J = 17.4 Hz, 1H), 4.48 - 4.33 (m, 3H), 3.61 - 3.46 (m, 7H), 2.99 - 2.80 (m, 5H), 2.73 (d, J = 4.9 Hz, 2H), 2.60 (ddd, J = 17.4, 4.4, 2.2 Hz, 1H), 2.43 (dd, J = 13.2, 4.5 Hz, 1H), 2.00 (dd, J = 9.3, 4.3 Hz, 1H); LC-MS m / z (ESI): 576.2 [M+1] + .

[0292] Example 30: Preparation of compound 30

[0293] Reference to the preparation method of Example 16, the raw material 1-benzyloxycarbonyl-3-pyrrolidinone is replaced by (R)-N-benzyloxycarbonyl-2-pyrrolidinecarboxaldehyde, V2 is replaced by the preparation of reference 4i, the raw material R-(+)-tert-butylsulfinamide is replaced by the intermediate prepared by S-(-)-tert-butylsulfinamide to prepare the title product compound 30 (26.70 mg, 30.27% yield). 1 H NMR (400 MHz, DMSO-d6) δ 9.74 (d, J = 4.9 Hz, 1H), 8.45 (s, 1H), 7.99 (dd, J = 7.7, 4.3 Hz, 1H), 7.69 - 7.60 (m, 2H), 7.54 (s, 1H), 7.46 (d, J = 7.9 Hz, 1H), 7.17 (d, J = 3.7 Hz, 2H), 5.08 (dd, J = 13.3, 5.1 Hz, 1H), 4.87 (t, J = 7.1 Hz, 1H), 4.42 (dd, J = 17.3, 4.9 Hz, 1H), 4.27 (dd, J = 17.3, 2.1 Hz, 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]+ .

[0294] Example 31: Preparation of compound 31

[0295] The title product compound 31 (140 mg, 73.33% yield) was prepared by following the procedure of Example 27, replacing the starting material methylamine with 2- (methylsulfonyl) ethylamine. 1 H NMR (400 MHz, DMSO-d6) δ 11.00 (s, 1H), 8.61 (d, J = 3.5 Hz, 1H), 8.15 (s, 1H), 7.70 (d, J = 7.9 Hz, 1H), 7.62 (d, J = 2.1 Hz, 1H), 7.55 (s, 1H), 7.48 (d, J = 7.9 Hz, 1H), 7.20 (d, J = 8.4 Hz, 1H), 7.10 (dd, J = 8.4, 2.2 Hz, 1H), 6.88 (dd, J = 7.8, 2.9 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.89 (p, J = 6.9 Hz, 1H), 4.45 (dd, J = 17.5, 2.0 Hz, 1H), 4.31 (d, J = 17.3 Hz, 1H), 3.63 - 3.44 (m, 4H), 3.25 (t, J = 6.9 Hz, 2H), 3.00 (d, J = 7.6 Hz, 5H), 2.96 - 2.80 (m, 3H), 2.75 (t, J = 5.4 Hz, 2H), 2.59 (d, J = 17.4 Hz, 1H), 2.45 - 2.28 (m, 1H), 1.98 (ddd, J = 9.9, 5.2, 2.6 Hz, 1H), 1.41 (d, J = 6.9 Hz, 3H); LC-MS m / z (ESI): 634.2 [M+1] + .

[0296] Example 32: Preparation of compound 32

[0297] The title product compound 32 (54 mg, 30.32% yield) was prepared by following the procedure of Example 27, replacing the starting material methylamine with isopropylamine. 1H NMR (400 MHz, DMSO-d6) δ 10.99 (s, 1H), 9.22 (s, 1H), 8.36 (s, 1H), 7.67 (d, J = 7.7 Hz, 1H), 7.62 (s, 1H), 7.53 (s, 1H), 7.46 (d, J = 7.9 Hz, 2H), 7.16 (t, J = 8.3 Hz, 2H), 5.09 (dd, J = 13.4, 4.7 Hz, 1H), 4.92 - 4.84 (m, 1H), 4.43 (d, J = 17.7 Hz, 1H), 4.28 (d, J = 16.8 Hz, 1H), 3.52 (dd, J = 15.0, 7.8 Hz, 4H), 2.89 (s, 1H), 2.81 (dd, J = 15.0, 7.8 Hz, 4H), 2.58 (dd, J = 36.5, 19.1 Hz, 2H), 2.41 - 2.28 (m, 1H), 1.98 (s, 1H), 1.38 (d, J = 6.9 Hz, 3H), 1.01 (d, J = 6.0 Hz, 6H). LC-MS m / z (ESI): 570.2 [M+1] + .

[0298] Example 33: Preparation of compound 33

[0299] The title product compound 33 (231 mg, 43.20% yield) was prepared by following the procedure of Example 27, replacing the raw material methylamine with ethylamine. 1 H NMR (400 MHz, DMSO-d6) δ 10.99 (br s, 1H), 9.36 (s, 1H), 8.38 (s, 1H), 7.66 (d, J = 7.9 Hz, 1H), 7.62 (s, 2H), 7.54 (s, 1H), 7.46 (d, J = 7.8 Hz, 1H), 7.16 (s, 2H), 5.09 (dd, J = 13.3, 5.1 Hz, 1H), 4.87 (t, J = 7.0 Hz, 1H), 4.42 (dd, J = 17.2, 4.0 Hz, 1H), 4.28 (d, J = 17.3 Hz, 1H), 3.55 (t, J = 5.6 Hz, 5H), 2.89 (d, J = 4.9 Hz, 3H), 2.83 (t, J = 6.7 Hz, 2H), 2.78 - 2.73 (m, 2H), 2.57 (d, J = 17.4 Hz, 1H), 2.40 - 2.31 (m, 1H), 2.01 - 1.92 (m, 1H), 1.38 (d, J = 6.9 Hz, 3H), 1.07 (t, J = 6.5 Hz, 3H). LC-MS m / z (ESI): 556.2 [M+1] + .

[0300] Example 34: Preparation of compound 34

[0301] The title product compound 34 (32 mg, 28.23% yield) was prepared according to the procedure described in Example 27 by replacing the starting material methylamine with (S)-3-aminotetrahydrofuran. 1 H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 8.85 (s, 1H), 8.21 (s, 1H), 7.67 (d, J = 7.8 Hz, 1H), 7.61 (d, J = 1.8 Hz, 1H), 7.53 (s, 1H), 7.46 (d, J = 7.9 Hz, 1H), 7.17 (d, J = 8.4 Hz, 1H), 7.14 - 7.08 (m, 2H), 5.09 (dd, J = 13.3, 4.9 Hz, 1H), 4.91 - 4.84 (m, 1H), 4.43 (dd, J = 17.4, 2.3 Hz, 1H), 4.29 (d, J = 17.3 Hz, 1H), 3.73 (dd, J = 14.8, 7.6 Hz, 1H), 3.67 - 3.63 (m, 1H), 3.60 (dd, J = 7.9, 6.1 Hz, 1H), 3.54 (dd, J = 12.6, 5.6 Hz, 2H), 3.48 - 3.43 (m, 3H), 3.39 (d, J = 10.7 Hz, 1H), 2.90 (dd, J = 10.6, 6.6 Hz, 1H), 2.82 (t, J = 6.9 Hz, 2H), 2.73 (dt, J = 12.5, 6.1 Hz, 2H), 2.58 (d, J = 17.1 Hz, 1H), 2.37 (d, J = 13.0 Hz, 1H), 2.01 - 1.92 (m, 2H), 1.66 (dd, J = 11.1, 5.7 Hz, 1H), 1.39 (d, J = 6.9 Hz, 3H). LC-MS m / z (ESI): 598.0 [M+1] + .

[0302] Example 35: Preparation of compound 35

[0303] The title product compound 35 (102.83 mg, 39.52% yield) was prepared according to the procedure described in Example 27 by replacing the starting material methylamine with (R)-3-aminotetrahydrofuran. 1H NMR (400 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.73 (d, J = 3.8 Hz, 1H), 8.19 (s, 1H), 7.68 (d, J = 7.8 Hz, 1H), 7.61 (d, J = 2.1 Hz, 1H), 7.54 (s, 1H), 7.47 (d, J = 7.9 Hz, 1H), 7.18 (d, J = 8.4 Hz, 1H), 7.09 (dd, J = 8.4, 2.2 Hz, 1H), 7.00 (dd, J = 7.7, 3.3 Hz, 1H), 5.09 (dd, J = 13.3, 5.1 Hz, 1H), 4.88 (p, J = 6.9 Hz, 1H), 4.43 (dd, J = 17.3, 2.8 Hz, 1H), 4.29 (d, J = 17.3 Hz, 1H), 3.71 (q, J = 7.5 Hz, 1H), 3.65 (dd, J = 9.0, 5.9 Hz, 1H), 3.59 (td, J = 8.0, 5.7 Hz, 1H), 3.52 (t, J = 7.0 Hz, 2H), 3.45 (t, J = 5.5 Hz, 2H), 3.43 - 3.40 (m, 1H), 3.33 (td, J = 6.8, 3.8 Hz, 1H), 2.90 (ddd, J = 17.2, 13.6, 5.4 Hz, 1H), 2.82 (t, J = 7.0 Hz, 2H), 2.71 (dq, J = 11.8, 5.7 Hz, 2H), 2.62 - 2.54 (m, 1H), 2.44 - 2.29 (m, 1H), 2.02 - 1.87 (m, 2H), 1.68 - 1.58 (m, 1H), 1.39 (d, J = 6.9 Hz, 3H); LC-MS m / z (ESI): 598.3 [M+1] + .

[0304] Example 36: Preparation of compound 36

[0305] The title product compound 36 (285.97 mg, 41.69% yield) was prepared by following the procedure of example 27, replacing the starting material methylamine with 2-aminoethyl sulfonamide. 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.58 (s, 1H), 8.16 (s, 1H), 7.70 (d, J = 7.9 Hz, 1H), 7.62 (d, J = 2.2 Hz, 1H), 7.55 (s, 1H), 7.48 (d, J = 7.9 Hz, 1H), 7.20 (d, J = 8.4 Hz, 1H), 7.10 (d, J = 8.8 Hz, 1H), 6.86 (s, 2H), 5.10 (dd, J = 13.3, 5.0 Hz, 1H), 4.90 (t, J = 7.1 Hz, 1H), 4.45 (d, J = 17.3 Hz, 1H), 4.31 (d, J = 17.3 Hz, 1H), 3.53 (t, J = 7.1 Hz, 2H), 3.45 (t, J = 5.5 Hz, 2H), 3.34 - 3.30 (m, 1H), 3.10 (d, J = 7.0 Hz, 2H), 2.92 (s, 3H), 2.83 (t, J = 7.1 Hz, 2H), 2.68 (s, 2H), 2.60 (d, J = 16.9 Hz, 1H), 2.39 (d, J = 14.0 Hz, 1H), 2.00 (s, 1H), 1.42 (d, J = 6.9 Hz, 3H); LC-MS m / z (ESI): 635.2 [M+1] + .

[0306] Biological test

[0307] Test Example 1: Test of compound on MDA-MB-453 cell proliferation inhibition activity

[0308] The proliferation inhibition activity on human breast ductal carcinoma cells MDA-MB-453 was tested by taking the compounds of the examples of the present application and control compound a as examples, to evaluate the in vitro cell activity of the compounds, wherein the structure of control compound a is shown below, which can be prepared by referring to the existing literature or obtained by purchase.

[0309] Control compound a

[0310] I. Reagents required for the experiment

[0311] II. Plates and instruments required for the experiment

[0312] III. Experimental method

[0313] 1. MDA-MB-453 cells in logarithmic phase were seeded into 384 cell culture plates except for negative control wells, to obtain compound test wells and positive control wells, with 45 μL of IMDM medium containing 10% FBS, at a cell density of 2000 / well, and the negative control wells were 45 μL of IMDM medium containing 10% FBS without cells;

[0314] 2. The required compound test final concentration solution was prepared by 10 times dilution with 100% DMSO;

[0315] 3. 5 μL of the prepared compound solution was added to the compound test wells, and 5 μL of 2.5% DMSO solution was added to the negative control wells and the positive control wells;

[0316] 4. The cell culture plate was placed in a 37°C, 5% CO2 cell incubator for 3 days;

[0317] 5. After taking out from the incubator, it was placed at room temperature for 0.5 h, and the cell culture plate was allowed to return to room temperature;

[0318] 6. The CTG reagent was taken out from the refrigerator in advance and allowed to return to room temperature;

[0319] 7. 40 μL of the CTG reagent was added to each well of the cell culture plate;

[0320] 8. The cell culture plate was placed at room temperature and incubated in the dark for 0.5 h;

[0321] 9. The luminescence signal value was read using a Tecan enzyme label meter.

[0322] Four, data calculation

[0323] 1. The PC well is the positive control well, i.e., the maximum value well tested on day 3, and the NC well is the negative control well, i.e., the minimum value well tested on day 3.

[0324] 2. Calculation of compound inhibition rate

[0325] wherein Inhibition ratio represents the inhibition rate of the tested compound, Signal Test represents the signal value of the cells after adding the detection compound for 72 h, SignalMean of NC represents the signal value of the negative control well, and SignalMean of PC represents the signal value of the positive control well. The XLFIT 5.0 software (UK IDBS Company) was used for fitting, taking the logarithmic value of the compound concentration as the X axis and the inhibition rate as the Y axis, and the four-parameter model was used to calculate the half-inhibitory concentration IC 50 .

[0326] The test results of the compound of the embodiment of the present application on the inhibition activity of MDA-MB-453 cell proliferation are shown in Table 1.

[0327] Table 1

[0328] The test results show that the compound of the present application has obviously improved inhibition activity on MDA-MB-453 cells compared with the control compound a.

[0329] Test Example 2: Experiment of compound on GSPT1 protein degradation in MDA-MB-453 cells

[0330] I. Reagents

[0331] II. Consumables and instruments

[0332] III. Experimental process

[0333] 1. Before testing, use 20 μg / mL PDL solution to coat the 384-well culture plate, and then store the coated culture plate in a 4°C refrigerator after covering with sealing film;

[0334] 2. MDA-MB-453 cells were subcultured in complete medium (IMDM + 10% FBS) to maintain logarithmic growth. Before inoculation, the cells were collected and counted after washing with PBS and TE digestion, and inoculated in the coated 384-well culture plate at a density of 2,000 cells per well (including compound test wells, negative control wells and positive control wells, wherein the negative control wells do not contain cells, and the compound test wells and positive control wells contain cells), and then incubated in an incubator at 37°C, 5% CO2 for 4 hours;

[0335] 3. Prepare compound gradient dilutions with DMSO, add the gradient dilutions to the compound test wells, and add IMDM containing DMSO to the negative control wells and positive control wells, and the final concentration of DMSO in all wells is 0.25 vol%. Then, continue to incubate in an incubator at 37°C, 5% CO2 for 24 hours;

[0336] 4. Take out the culture plate, directly add fixing solution (2.5 vol% GA in PBS) to all wells, and fix the cells at room temperature for 30 minutes, and then discard the culture solution;

[0337] 5. Add calibration staining solution (40 μg / mL Hoechst 33342 and 0.1 vol% Triton X-100 in PBS) to all wells, and stain at room temperature for 30 minutes in the dark, and then discard the staining solution, and immediately read the fluorescence signal value on the enzyme marker (excitation wavelength360nm / emission wavelength 460nm );

[0338] 6. Add antigen retrieval solution (1 mM EDTA in water) to all wells, and place the microplate on a microplate shaker heater for high temperature retrieval for 2 hours. Then remove the plate and discard the retrieval solution after it has cooled to room temperature.

[0339] 7. Prepare antibody diluent (1 wt% BSA in TBST) and add to the negative control wells. Then add GSP T1 antibody to the antibody diluent at a ratio of 1:1000 and add to the remaining wells, except the negative control wells. Cover the plate with adhesive film and incubate at 4°C overnight. Then discard all liquid.

[0340] 8. Wash all wells with TBST solution three times. Prepare antibody diluent (1 wt% BSA in TBST) and add anti-rabbit IgG-HRP conjugated antibody to the antibody diluent at a ratio of 1:500 and add to all wells. Incubate at room temperature for 2 hours. Then discard all liquid.

[0341] 9. Wash all wells with TBST solution three times. Then add TMB substrate to all wells and incubate at room temperature for 20 minutes in the dark. Immediately after incubation, add 2M sulfuric acid to stop the color development reaction. Read the signal on a microplate reader: absorbance value (OD 450 nm ).

[0342] Four, Data calculation

[0343] 1. Before analysis, all data will be normalized according to the signal in each well, as follows:

[0344] Where TMB values (OD 450 nm ) represent the absorbance value of the cells produced, please refer to experiment operation 9, Hoechst 33342 values (Fluorescence 460 nm ) represent the fluorescence signal value of the cells, please refer to experiment operation 5, Max (Hoechst 33342 value) represents the fluorescence signal value of the positive control well cells.

[0345] 2. The calculation method of compound degradation rate is as follows:

[0346] Where DR% represents the degradation rate of the test compound to the target protein, Signal TestThe 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.

[0347] 3. Fit the compound concentration and DR% using a four-parameter model, as follows:

[0348] DR% PC The percentage (DR%) indicates the degradation rate of the target protein in the positive control wells. NC The degradation rate of the target protein in the negative control wells is represented by LogCmpd, where 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 2 below:

[0349] Table 2

[0350] As can be seen from the results in the table above, the compounds in this application have strong GSPT1 protein degradation activity.

[0351] Test Example 3: Pfeiffer Cell Selectivity Experiment

[0352] I. Reagents

[0353] II. Consumables and Instruments

[0354] III. Experimental Procedure

[0355] 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.

[0356] 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.

[0357] 3. Add the gradient dilution of the compound prepared in DMSO to the compound test wells, and add the RPMI 1640 containing DMSO to the negative control wells and the positive control wells. The final concentration of DMSO in all wells is 0.1 vol%. Then, place the plate in an incubator at 37°C, 5% CO2, and continue incubation for 24 hours;

[0358] 4. Take out the plate and directly add the fixing solution (PBS containing 2.5 vol% GA) to all wells, and fix the cells at room temperature for 30 minutes. Then, discard the solution;

[0359] 5. Add the calibration staining solution (PBS containing 40 μg / mL Hoechst 33342 and 0.1 vol% Triton X-100) to all wells, and stain at room temperature for 30 minutes in the dark. Then, discard the staining solution, and immediately read the fluorescence signal value (excitation wavelength 360nm / emission wavelength 460nm ) on the microplate reader;

[0360] 6. Add the antigen repair solution (1x boric acid buffer solution) to all wells, and place the plate in a microplate shaking heater for high-temperature repair for 2 hours. Then, take out the plate and discard the repair solution after cooling to room temperature;

[0361] 7. Prepare the antibody dilution solution (TBST containing 1 wt% BSA), and add it to the negative control wells. Then, add the GSPT1 antibody (or IKZF1 antibody, or IZKF3 antibody, or CK1a antibody) to the antibody dilution solution at a ratio of 1:1000, and add it to the remaining wells except the negative control wells. After covering with sealing film, incubate at 4°C overnight, and then discard all the liquid;

[0362] 8. Wash all wells with TBST solution for three times. Prepare the antibody dilution solution (TBST containing 1 wt% BSA), and add the anti-rabbit IgG-HRP conjugated antibody to the antibody dilution solution at a ratio of 1:500, and add it to all wells. Incubate at room temperature for 2 hours, and then discard all the liquid;

[0363] 9. Wash all wells with TBST solution for three times. Then, add the TMB substrate to all wells, and incubate at room temperature in the dark for 20 minutes for color development. Immediately after the incubation, add 2M sulfuric acid to stop the color development reaction, and read the signal on the microplate reader: absorbance value (OD 450nm ).

[0364] Four, data calculation

[0365] 1. Before analysis, all data will be normalized according to the signal in each well, as follows:

[0366] TMB values (OD 450nm ) represent the absorbance values generated by the cells, please refer to Experimental Operation 9, Hoechst 33342 values (Fluorescence 460nm ) represent the fluorescence signal values of the cells, please refer to Experimental Operation 5, Max (Hoechst 33342 value) represents the fluorescence signal values of the cells in the positive control well.

[0367] 2. The degradation rate of the compound is calculated as follows:

[0368] wherein DR% represents the degradation rate of the target protein by the test compound, Signal Test represents the signal generated by the cells treated with the compound, SignalMean of NC represents the signal generated by the cells in the negative control well, and SignalMean of PC represents the signal generated by the cells in the positive control well.

[0369] 3. The four-parameter model is used to fit the compound concentration and DR% as follows:

[0370] wherein DR% PC represents the degradation rate of the target protein by the positive control well, DR% NC represents the degradation rate of the target protein by the negative control well, LogCmpd represents the logarithmic value of the compound concentration, and Hillslope represents the slope value obtained by curve fitting. The DC 50 values of different compounds are calculated by this model and compared. The test results are shown in Table 3.

[0371] Table 3: Inhibition activity of compounds on IKZF1 and IKZF3

[0372] The test results show that the compounds of the present application have more excellent selectivity for IKZF1 and IKZF3 compared with the control compounds a and b, wherein the structure of the control compound b is shown below, which can be prepared by referring to the existing literature or obtained by purchase.

[0373] Control compound b

[0374] Test Example 4: In vivo pharmacokinetic test in mice

[0375] The drug concentration in the plasma of mice at different time points after intravenous administration (IV) of the compound of the present application was determined by LC / MS / MS method, the pharmacokinetic behavior of the compound of the present application in mice was studied, and the pharmacokinetic characteristics were evaluated.

[0376] Reagents

[0377] 1. Experimental Protocol: Healthy adult male ICR mice, weighing 25-40 g, 3 per group, all mice were free to eat and drink, animals were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. or Shanghai Jihui Experimental Animal Breeding Co., Ltd. The animals were transported to the animal room for at least 3 days of acclimation, SPF area room, indoor temperature maintained at 20-26℃, indoor relative humidity maintained at 40-70%. Daylight lighting, 12 hours of light (08:00-20:00) and 12 hours of no light, 2-5 per cage (same drug administration group). Unlimited access to feed (irradiated and sterilized, purchased from Shaanxi Qinle Pharmaceutical Chemical Co., Ltd., China) and water (tap water purified by ultrapure water filtration system). The specific dosing regimen is as follows:

[0378] 2. Animal Experiment Prescription:

[0379] Solvent: 5% DMSO / 10% Solutol HS15 / 85% water; Preparation of drug preparation: accurately weigh the appropriate amount of test compound, add 5% of the formula volume of DMSO, shake and ultrasonic to fully dissolve, after dissolving, add 10% of the formula volume of Solutol HS15, vortex for 2 minutes, finally add 85% of the formula volume of pure water, ultrasonic to get a clear and transparent solution with a concentration of 0.4 mg / mL.

[0380] 3. Dosing method: select animals that meet the experimental requirements before dosing, weigh and label. ICR mice were administered 2 mg / kg via tail vein.

[0381] 4. Sample collection: before sampling, bind the mice, at the predetermined blood sampling time points, each dosed mouse was sampled at 0.083, 0.25, 0.5, 1, 2, 4, 7, 24 h after dosing, a total of 8 time points, about 80 μL of blood was collected through the submandibular vein. The blood was transferred to a 1.5 mL tube pre-added with K2EDTA, centrifuged (eppendorf, Centrifuge 5804R) for 3 min (10000 rpm, 4℃), the plasma was separated, the whole process was completed within 15 min after blood sampling. All samples need to be stored in a -20℃ refrigerator (Panasonic) until sample analysis.

[0382] 5. Sample analysis and calculation: LC / MS / MS (SCIEX, TRIPLE QUAD 6500+) method was used to determine the drug concentration and Data Analysie System software (Shanghai Boji Pharmaceutical Technology Co., Ltd., version number 3.0) was used to calculate the pharmacokinetic parameters such as drug half-elimination time T 1 / 2 , drug exposure AUC 0-t , drug clearance CLz, etc. The pharmacokinetic property parameters of some compounds of the present application in mice under the same dose and administration mode are shown in Table 4:

[0383] Table 4 Pharmacokinetic parameters of compounds in mice

[0384] From the data in the above table, it can be seen that the representative compounds of the present application have more excellent in vivo pharmacokinetic parameter properties compared with the control compound a.

[0385] All the documents mentioned in the present application are cited as references in the present application, just as each document is cited as a reference individually. In addition, it should be understood that various modifications or changes can be made to the present application by those skilled in the art after reading the above description of the present application, and these equivalent forms also fall within the scope defined by the claims of the present application.

[0386] The technical features of the above-described embodiments can be combined in any manner. In order to make the description concise, all possible combinations of the technical features in the above-described embodiments are not described, however, as long as the combinations of the technical features do not contradict each other, they should be considered as falling within the scope of the present application.

[0387] The above-described embodiments only express several implementation manners of the present application, and the description is relatively specific and detailed, however, it should not be understood as a limitation on the scope of the patent. It should be pointed out that for those skilled in the art, several modifications and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application. Therefore, the protection scope of the patent of the present application should be subject to the appended claims.

Claims

A compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: wherein R1is hydrogen or halogen; R2, R3are each independently hydrogen or C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl); and R1, R2, R3are not simultaneously hydrogen; n is 1, 2 or 3; R 41 , R 42 , R 43 , R 44 each independently is hydrogen or halogen; R5, R6, R a , R b , R c , R d , R e , R f , m is selected from the group consisting of: (i) R5, R6are each independently hydrogen, C 1-8 alkyl, C 3-6 cycloalkyl, -C 1-4 alkylene-C 3-6 cycloalkyl, halogen-C 1-8 alkyl, 4- to 6-membered heterocycloalkyl or -C 1-4 alkylene-R7, said C 1-8 alkyl, C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen, C 1-8 alkyl; R7is C 3-6 cycloalkyl, -S(O)2-C 1-4 alkyl, -S(O)-C 1-4 alkyl, -S(O)2-NH2, -C(O)NH2, hydroxy or cyano; said C 3-6 cycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen; 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) R6is 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 heterocycloalkyl, which C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen, 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 R5is selected from the group consisting of hydrogen; C1-6alkyl; C1-6alkyl substituted with 1, 2, or 3 substituents independently selected from the group consisting of halogen; and a saturated 4- to 7-membered nitrogen-containing heterocycloalkyl ring formed with R5; said saturated 4- to 7-membered nitrogen-containing heterocycloalkyl ring being unsubstituted or substituted with 1, 2, or 3 substituents independently selected from the group consisting of C1-6alkyl, halogen; 1-8 alkyl, halogen; m is 0, 1 or 2; (iii) R6is 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 heterocycloalkyl, which C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen, 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 with R5to form a saturated 4- to 7-membered nitrogen-containing heterocycloalkyl; said saturated 4- to 7-membered nitrogen-containing heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of C 1-8 alkyl, halogen; m is 0, 1 or 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R1is halogen; R2, R3are each independently hydrogen. The compound of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R1is fluorine; R2, R3are each independently hydrogen. The compound of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R1is hydrogen; R2is hydrogen; R3is C 1-3 alkyl. The compound of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R5is hydrogen, C 1-3 alkyl, C 3-6 cycloalkyl, -C 1-2 alkylene-C 3-6 cycloalkyl, halogen-C 1-3 alkyl (preferably fluorinated C 1-3 alkyl), 4- to 6-membered heterocycloalkyl or -C 1-2 alkylene-R7, said C 1-3 alkyl, C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen, C 1-3 alkyl; R6is hydrogen; R7is C 3-6 cycloalkyl, -S(O)2-C 1-3 alkyl, -S(O)-C 1-3 alkyl, -S(O)2-NH2, -C(O)NH2, hydroxy or cyano; said C 3-6 cycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen; 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 of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R6is hydrogen, C 1-3 alkyl, C 3-6 cycloalkyl, -C 1-2 alkylene-C 3-6 cycloalkyl, halo-C 1-3 alkyl, or 4- to 6-membered heterocycloalkyl, said C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen, 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 R5is selected from the group consisting of hydrogen; C1-6alkyl; C1-6alkyl substituted with 1, 2, or 3 substituents independently selected from the group consisting of halogen; and a saturated 4- to 7-membered nitrogen-containing heterocycloalkyl ring formed with R5; said saturated 4- to 7-membered nitrogen-containing heterocycloalkyl ring being unsubstituted or substituted with 1, 2, or 3 substituents independently selected from the group consisting of C1-6alkyl, halogen; 1-8 alkyl, halogen; m is 0 or 1. The compound of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R6is hydrogen, C 1-3 alkyl, C 3-6 cycloalkyl, -C 1-2 alkylene-C 3-6 cycloalkyl, halogen-C 1-3 alkyl, or 4- to 6-membered heterocycloalkyl, which C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen, 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 R5is selected from the group consisting of hydrogen; C1-6alkyl; C1-6alkyl substituted with 1, 2, or 3 substituents independently selected from the group consisting of halogen; and a saturated 4- to 7-membered nitrogen-containing heterocycloalkyl ring formed with R5; said saturated 4- to 7-membered nitrogen-containing heterocycloalkyl ring being unsubstituted or substituted with 1, 2, or 3 substituents independently selected from the group consisting of C1-6alkyl, halogen; 1-8 alkyl, halogen; m is 0. The compound of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein The compound of formula (I) is represented by formula (I-A) or formula (I-B): wherein R 51 , R 62 each independently is hydrogen, C 1-3 alkyl, C 3-6 cycloalkyl, -C 1-2 alkylene-C 3-6 cycloalkyl, halo-C 1-3 alkyl, 4- to 6-membered heterocycloalkyl or -C 1-2 alkylene-R7, said C 1-3 alkyl, C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group of halogen, C 1-3 alkyl; R7is C 3-6 cycloalkyl, -S(O)2-C 1-3 alkyl, -S(O)-C 1-3 alkyl, -S(O)2-NH2, -C(O)NH2, hydroxy or cyano; said C 3-6 cycloalkyl is unsubstituted or substituted by 1, 2 or 3 substituents selected from the group consisting of halogen; n is 1; s is 1, 2 or 3; t is 0, 1 or 2; and when s is 1, t is not 0; R0is each independently hydrogen, C 1-8 alkyl, halogen; q is 0, 1, 2 or 3; p is 0, 1 or 2. The compound of claim 1 or 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R1is fluorine; R2, R3are each independently hydrogen. The compound of claim 1 or 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R1is hydrogen; R2is hydrogen; R3is C 1-3 alkyl. The compound of claim 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R 51 a group selected from the group consisting of -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 -halogenocyclopropyl (preferably -(CH2) 0-1 -fluorocyclopropyl), -(CH2) 0-1 -halogenocyclobutyl (preferably -(CH2) 0-1 -fluorocyclobutyl), -(CH2) 0-1 -halogenocyclopentyl (preferably -(CH2) 0-1 -fluorocyclopentyl), -(CH2) 0-1 -halogenocyclohexyl (preferably -(CH2) 0-1 -fluorocyclohexyl), -(CH2) 0-1 -cyclopropyl, -(CH2) 0-1 -cyclobutyl, -(CH2) 0-1 -cyclopentyl, -(CH2) 0-1 -cyclohexyl, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, oxazolidinyl, dioxolanyl, piperidinyl, piperazinyl, morpholinyl, dioxanyl, thiomorpholinyl, thiomorpholine-1,1 -dioxide, tetrahydropyranyl. The compound of claim 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R 62 is hydrogen. The compound of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein The compound of formula (I) is selected from any one of the following structures: The compound of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein The compound of formula (I) is selected from any one of the following structures: A pharmaceutical composition, characterized in that, A pharmaceutical composition comprising a compound of any one of claims 1-14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof; and a pharmaceutically acceptable carrier. Use of a compound of any one of claims 1-14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition of claim 15, in the manufacture of a medicament for treating or preventing a disease associated with a mutation, an imbalance in expression, an allosteric change and a dysfunction of GSPT1, N-MYC or C-MYC protein. The use according to claim 16, characterized in that The disease is selected from the group consisting of 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 cancer, non-Hodgkin's lymphoma, central nervous system tumor, prostate cancer, thyroid cancer, acute myeloid leukemia, myelodysplastic syndrome. Use of a compound of any one of claims 1-14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition of claim 15, in the manufacture of a GSPT1 degrader. Use of a compound of any one of claims 1-14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition of claim 15, in the manufacture of a medicament for treating a disease associated with or caused by GSPT1. Use according to claim 19, characterized in that The disease is a cancer associated with GSPT1 selected from the group consisting of glioma, thyroid cancer, lung cancer, colorectal cancer, head and neck cancer, gastric cancer, liver cancer, pancreatic cancer, kidney cancer, urothelial cancer, prostate cancer, testicular cancer, breast cancer, cervical cancer, endometrial cancer, ovarian cancer, melanoma, multiple myeloma, acute myeloid leukemia, myelodysplastic syndrome.

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