Substituted tricyclic derivative as well as pharmaceutical composition and application thereof

Abstract

The invention discloses a substituted tricyclic derivative. The structure of the substituted tricyclic derivative is shown as a formula (I). In addition, the invention also discloses a pharmaceutical composition of the derivative and application thereof. The compound provided by the invention has a remarkable inhibition effect and a degradation effect on SHP2, has excellent cell inhibition activity, and has a relatively high application value. # imgabs0 #

Description

[0001] This application claims the priority of the Chinese patent application filed with the China Patent Office on July 6, 2023, with application number CN202310827216.4 and invention name “Substituted tricyclic derivatives, pharmaceutical compositions and uses thereof”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The present application relates to the field of medical technology, and in particular to a substituted tricyclic derivative, a pharmaceutically acceptable salt thereof, a pharmaceutical composition and its medical use. Background Art

[0003] Protein tyrosine phosphatases (PTPs) catalyze the dephosphorylation of phosphotyrosine and are key control elements in mammalian signal transduction. Deviations in their biological functions can cause regulatory disorders in the body and lead to the occurrence of a variety of diseases such as cancer, diabetes, and autoimmune diseases. Among them, Src homology 2 domain-containing protein tyrosine phosphatase (SHP2) is currently the only confirmed proto-oncoprotein in the PTP family. Src homology-2 phosphatase (SHP2) is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene that contributes to a variety of cellular functions including proliferation, differentiation, cell cycle maintenance, and migration. SHP2 is involved in signal transduction through the Ras-mitogen-activated protein kinase (JAK-STAT or phosphoinositide 3-kinase-AKT pathway).

[0004] SHP2 is considered to be an ideal target for cancer intervention, mainly in three aspects: First, SHP2 is a common node for multiple RAS signaling pathways. Activating RAS is very important for the growth and survival of cancer cells. Almost all protein tyrosine kinases (PTKs) activate the RAS signaling pathway by activating SHP2. Therefore, a suitable SHP2 inhibitor can "catch all" different PTK gene mutations and has the potential to become a broad-spectrum anti-cancer drug.

[0005] Secondly, due to the overlap of PTK and SHP2 signaling pathways, SHP2 inhibitors can be used in combination with kinase inhibitors to dually inhibit interconnected signaling pathways. This combination therapy is more effective than single therapy, is not easy to develop drug resistance, and can reverse the acquired drug resistance of PTK inhibitors; finally, SHP2 is also involved in the signaling pathway of programmed cell death checkpoint PD-1 / PD-L1, and is a direct downstream complex element of PD-1 / PD-L1. Inhibiting the binding of SHP2 to PD-1 may restore T cell activation, opening up a new path for this target in immunotherapy.

[0006] Mutations in the PTPN11 gene and subsequently SHP2 have been identified in a variety of human diseases, such as Noonan syndrome, Leopard syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, acute myeloid leukemia, and breast, lung, and colon cancers. Therefore, SHP2 represents a highly attractive target for the development of new therapeutics for a variety of diseases. Summary of the invention

[0007] The purpose of the present application is to provide a novel bifunctional compound which has an activity inhibitory and degradation effect on SHP2.

[0008] The first aspect of the present application provides a compound represented by formula (I), or a pharmaceutically acceptable salt thereof:

[0009]

[0010] in,

[0011] R1 and R2 are each independently hydrogen, C 1-3 Alkyl, -NH2, C 1-3 Alkoxy, cyano, hydroxy, carboxyl, halogen (preferably fluorine or chlorine), halogenated C 1-3 Alkyl, halogenated C 1-3 Alkoxy, -C(O)NH2, -C(O)C 1-3 Alkyl, -OC(O)C 1-3 Alkyl or -C(O)OC 1-3 alkyl;

[0012] R3 and R4 are each independently hydrogen, C 1-3 Alkyl or -C(O)C 1-6 alkyl;

[0013] (R5) m1 represents m1 R5, m1 is 0 or 1, each R5 is the same or different, and each independently represents halogen (preferably fluorine, chlorine or bromine), cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, -NH2, -NHCOC 1-3 Alkyl, -C(O)C 1-3 Alkyl, -C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, hydroxyl, carboxyl, halogenated C 1-3 Alkyl or halogenated C 1-3 Alkoxy;

[0014] (R6) m2represents m2 R6, m2 is 0, 1, 2, 3 or 4, each R6 is the same or different, and each independently represents halogen (preferably fluorine, chlorine or bromine), cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl or halogenated C 1-3 Alkoxy;

[0015] W is -CH2- or -C(O)-;

[0016] X is -(X1) p1 -(L1) q1 -(X2) p2 -(L2) q2 -(X3) p3 -(L3) q3 -(X4) p4 -(L4) q4 -,

[0017] Wherein, L1, L2, L3, and L4 are each independently C 1-10 Alkylene;

[0018] X1, X2, X3, and X4 are each independently -O-, -NH-, -C(O)-, -NHC(O)-, or -(OCH2CH2) p5 -, azetidine ring group, tetrahydropyrrole ring group, piperazine ring group, piperidine ring group, triazole ring group, C 2-4 Alkenylene, C 2-4 Alkyne group,

[0019] p1, p2, p3, p4, p5, q1, q2, q3, q4 are each independently 0, 1, 2, 3 or 4.

[0020] In some embodiments, R1 and R2 are each independently hydrogen, methyl, -NH2, methoxy, cyano, hydroxyl, carboxyl, fluorine, chlorine, trifluoromethoxy, difluoromethoxy, monofluoromethoxy, trifluoromethyl, difluoromethyl, monofluoromethyl, -C(O)NH2, -COCH3, -COOCH3 or -OCOCH3.

[0021] In some embodiments, R3 and R4 are each independently hydrogen, methyl or -C(O)CH3.

[0022] In some embodiments, (R5) m1It represents m1 R5, m1 is 0 or 1, each R5 is the same or different, and each independently represents fluorine, chlorine, cyano, methyl, methoxy, vinyl, ethynyl, -NH2, -NHCOCH3, -C(O)CH3, -C(O)OCH3, -OC(O)CH3, hydroxyl, carboxyl, trifluoromethyl, difluoromethyl, monofluoromethyl, trifluoromethoxy, difluoromethoxy or monofluoromethoxy.

[0023] In some embodiments, (R6) m2 represents m2 R6, m2 is 0, 1, 2, 3 or 4, each R6 is the same or different, and each independently is fluorine, chlorine, cyano, methyl, methoxy, vinyl, ethynyl, trifluoromethyl, difluoromethyl, monofluoromethyl, trifluoromethoxy, difluoromethoxy or monofluoromethoxy.

[0024] In some embodiments, the compound represented by formula (I) has the structure represented by formula (I-1),

[0025]

[0026] In some embodiments, m1 is 0.

[0027] In some embodiments, m1 is 1, and R5 is -NH2.

[0028] In some embodiments, R6 is halogen, preferably fluorine, chlorine or bromine.

[0029] In some embodiments, m2 is 2, and R6 is fluoro.

[0030] In some embodiments, R3, R4 are hydrogen.

[0031] In some embodiments, R1 is hydrogen.

[0032] In some embodiments, R2 is -NH2.

[0033] In some embodiments, W is -CH2-.

[0034] In some embodiments, W is -C(O)-.

[0035] In some embodiments, the structure Select from the following structures:

[0036]

[0037] In some embodiments, the structure Select from the following structures:

[0038]

[0039] In some embodiments, the structure Select from the following structures:

[0040]

[0041] In some embodiments, the compound of formula (I) is selected from the structure shown below,

[0042]

[0043] In some embodiments, L1, L2, L3, L4 are each independently -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, or -(CH2)7-.

[0044] In some embodiments, X1, X2, X3, and X4 are each independently -O-, -NH-, -C(O)-, -NHC(O)-, -(OCH2CH2)1-, -(OCH2CH2)2-, -(OCH2CH2)3-, -(OCH2CH2)4-,

[0045] In some embodiments, X is

[0046] Wherein, n is independently an integer of 1-10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0047] In some embodiments, X is selected from the structure shown below,

[0048]

[0049] In some embodiments, X is selected from the structure shown below,

[0050]

[0051] in End and connect, End and connect.

[0052] In some embodiments, X is selected from the structure shown below,

[0053]

[0054]

[0055] in End and connect, End and connect.

[0056] In some embodiments, the compound of formula (I) is a compound of formula (II),

[0057]

[0058] Among them, X is

[0059] Wherein, n is independently an integer of 1-10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0060] In some embodiments, X is selected from the following structures,

[0061]

[0062] Wherein, n is independently an integer of 1-5, for example, 1, 2, 3, 4 or 5.

[0063] In some embodiments, X is selected from the following structures,

[0064]

[0065] in End and connect, End and connection; wherein for

[0066] In some embodiments, the compound of formula (II) is a structure shown in formula (II-1),

[0067]

[0068] In some embodiments, the compound of formula (I) is a compound of formula (III),

[0069]

[0070] Wherein, X is selected from the following structures,

[0071] Wherein, n is independently an integer of 1-10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0072] In some embodiments, X is selected from the following structures,

[0073] Wherein, n is independently an integer of 1-5, for example, 1, 2, 3, 4 or 5.

[0074] In some embodiments, X is selected from the following structures,

[0075] in End and connect, End and connection; wherein said for In some embodiments, the compound of formula (III) is a structure shown in formula (III-1),

[0076]

[0077] In some embodiments, the compound of Formula (I) is selected from the specific compounds in the Examples.

[0078] In some embodiments, the compound of formula (I) is selected from the compounds in Table 1:

[0079] Table 1

[0080]

[0081]

[0082]

[0083] The second aspect of the present application provides a pharmaceutical composition, which includes the compound described in the first aspect of the present application, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0084] The third aspect of the present application provides the use of the compound described in the first aspect of the present application, or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition described in the second aspect of the present application in the preparation of a medicament for treating diseases or conditions mediated by SHP2 or associated with abnormal SHP2 activity.

[0085] In some embodiments, diseases or conditions associated with aberrant SHP2 activity include solid tumors and hematological tumors.

[0086] In some embodiments, the SHP2-mediated disease or condition is cancer, including but not limited to: juvenile myelomonocytic leukemia (JMML), acute myeloid leukemia (AML), B-cell acute lymphoblastic leukemia (B-ALL), neuroblastoma, esophageal cancer, breast cancer, lung cancer, colon cancer, gastric cancer, head and neck cancer.

[0087] The fourth aspect of the present application provides a method for treating a disease or condition mediated by SHP2 or associated with abnormal SHP2 activity, the method comprising administering to a patient a therapeutically effective amount of the compound described in the first aspect of the present application, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in the second aspect of the present application.

[0088] In some embodiments, diseases or conditions associated with aberrant SHP2 activity include solid tumors and hematological tumors.

[0089] In some embodiments, the SHP2-mediated disease or condition is cancer, including but not limited to: juvenile myelomonocytic leukemia (JMML), acute myeloid leukemia (AML), B-cell acute lymphoblastic leukemia (B-ALL), neuroblastoma, esophageal cancer, breast cancer, lung cancer, colon cancer, gastric cancer, head and neck cancer.

[0090] Beneficial effects of this application:

[0091] The compound of formula (I) provided in the present application has significant SHP2 enzyme inhibition activity and SHP2 protein degradation activity, and has excellent cell inhibition activity, and is expected to be developed into a drug for treating and / or preventing diseases or conditions mediated by SHP2 or associated with abnormal SHP2 activity.

[0092] Of course, implementing any product or method of the present application does not necessarily require achieving all of the advantages described above at the same time. BRIEF DESCRIPTION OF THE DRAWINGS

[0093] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application, and for ordinary technicians in this field, other embodiments can also be obtained based on these drawings.

[0094] Figure 1 The graph shows the degradation results of SHP2 protein by the compound of formula (I). DETAILED DESCRIPTION

[0095] After extensive and in-depth research, the applicant unexpectedly discovered substituted tricyclic derivatives, which have significant SHP2 enzyme inhibition activity and SHP2 protein degradation activity, and have excellent cell inhibition activity. Therefore, this series of compounds is expected to be developed into drugs for treating and / or preventing diseases or conditions mediated by SHP2 or associated with abnormal SHP2 activity. On this basis, the inventor completed this application.

[0096] Definition of terms

[0097] In order to more clearly understand the technical content of this application, the terms of this application are further explained below.

[0098] "Alkyl" refers to straight-chain or branched saturated aliphatic hydrocarbon groups. 1-6 "Alkyl" refers to an alkyl group having 1 to 6 carbon atoms, preferably C 1-3 Alkyl; Non-limiting examples of alkyl groups include: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and various branched chain isomers thereof.

[0099] "Alkenyl" refers to a straight or branched unsaturated aliphatic hydrocarbon group having one or more carbon-carbon double bonds (C=C). 2-4 "Alkenyl" refers to an alkenyl group having 2 to 4 carbon atoms; non-limiting examples of alkenyl include ethenyl, propenyl, isopropenyl, n-butenyl, isobutenyl, and the like.

[0100] "Alkynyl" refers to straight-chain or branched unsaturated aliphatic hydrocarbon groups having one or more carbon-carbon triple bonds. 2-4 "Alkynyl" refers to an alkynyl group having 2 to 4 carbon atoms; non-limiting examples of alkynyl include ethynyl, propynyl, n-butynyl, isobutynyl, and the like.

[0101] "Alkylene" is divalent and requires two binding partners. Formally, the second valence is generated by removing a hydrogen atom from an alkyl group, such as -CH3 and -CH2-, -CH2CH3 and -CH2CH2- or -CHCH3, etc. In certain embodiments, C 1-10 Alkylene, more preferably C 1-8 Alkylene, more preferably C 1-6 Alkylene, most preferably C 1-3 Alkylene. For example, C1-3 Alkylene includes -CH2-, -(CH2)2-, -CH(CH3)-, -(CH2)3-, -CH(CH2CH3), -CH2CH(CH3)-, and -C(CH3)2-. In certain embodiments, the alkylene may be methylene, ethylene, propylene, 1-methylethylene, butylene, 1-methylpropylene, 1,1-dimethylethylene, 1,2-dimethylethylene, pentylene, 1,1-dimethylpropylene, 2,2-dimethylpropylene, 1,2-dimethylpropylene, 1,3-dimethylpropylene, and the like. In the absence of any further definition, the generic terms propylene, butylene, pentylene, hexylene, and the like are intended to refer to all conceivable isomeric forms having the corresponding number of carbon atoms, i.e., propylene includes 1-methylethylene, and butylene includes 1-methylpropylene, 2-methylpropylene, 1,1-dimethylethylene, and 1,2-dimethylethylene.

[0102] "Alkenylene" is composed of at least two carbon atoms, wherein at least two adjacent carbon atoms are linked together by a C-C double bond, and the carbon atom can only be part of a C-C double bond. Formally, in the alkylene defined above, two hydrogen atoms on adjacent carbon atoms are formally removed and the free valence is saturated to form a second bond, then the corresponding alkenylene is formed. In certain embodiments, alkenylene can be vinylene, propenylene, 1-methylvinylene, butenylene, 1-methylpropenylene, 1,1-dimethylvinylene, 1,2-dimethylvinylene, etc. In the absence of any further definition, the general terms propenylene, butenylene, etc. mean all conceivable isomeric forms with the corresponding number of carbon atoms, i.e., propenylene includes 1-methylpropenylene, and butenylene includes 1-methylpropenylene, 2-methylpropenylene, 1,1-dimethylvinylene and 1,2-dimethylvinylene. For one or more double bonds, alkenylene can optionally exist in cis or trans or E or Z form.

[0103] "Alkynylidene" refers to a group consisting of at least two carbon atoms, wherein at least two adjacent carbon atoms are linked together by a C-C triple bond. Formally, in the alkylene group defined above, two hydrogen atoms are removed from two adjacent carbon atoms, respectively, and the free valence is saturated to form two additional bonds, forming the corresponding alkynylene. In certain embodiments, the alkynylene group may be ethynylene, propynylene, 1-methylethynylene, butynylene, 1-methylpropynylene, 1,1-dimethylethynylene, 1,2-dimethylethynylene, etc. In the absence of any further definition, the general terms propynylene, butynylene, etc. mean all conceivable isomeric forms with the corresponding number of carbon atoms, i.e., propynylene includes 1-methylethynylene, and butynylene includes 1-methylpropynylene, 2-methylpropynylene, 1,1-dimethylethynylene, and 1,2-dimethylethynylene.

[0104] "Alkyleneoxy" refers to a divalent alkoxy group. In certain embodiments, C 1-10 Alkyleneoxy, more preferably C 1-8 Alkyleneoxy, more preferably C 1-6 Alkyleneoxy, most preferably C 1-3 Alkyleneoxy. In certain embodiments, the alkyleneoxy group can be -OCH2-, -OCH(CH3)CH2-, -OCH2CH2O-, -CH2CH2O-, etc. In the absence of any further definition, the general terms propyleneoxy, butyleneoxy, pentyleneoxy, hexyleneoxy, etc. mean all conceivable isomeric forms having the corresponding number of carbon atoms, i.e., propyleneoxy includes -O(CH2)3O-, -O(CH3)3-, -OCH2CH(CH3)-, -OC(CH3)2-, -OCH(CH3)CH2-, -OCH2CH(CH3)O-, -OC(CH3)2O-, and -OCH(CH3)CH2O-.

[0105] "Halogen" refers to fluorine, chlorine, bromine or iodine.

[0106] "Halo" means that one or more (eg, 1, 2, 3 or all) hydrogen atoms in a group are replaced by halogen.

[0107] "Haloalkyl" refers to an alkyl group substituted with one or more (e.g., 1, 2, 3 or all) halogens, wherein the alkyl group is as defined above. Examples of haloalkyl groups include, but are not limited to, monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, monobromoethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, and the like.

[0108] "Alkoxy" refers to -O-alkyl, wherein alkyl is as defined above. Non-limiting examples of alkoxy include methoxy, ethoxy, n-propoxy, isopropoxy, and the like.

[0109] "Haloalkoxy" refers to an alkoxy group substituted with one or more (e.g., 1, 2, 3, 4, or 5) halogens, wherein the definition of alkoxy is as described above. Haloalkoxy includes, but is not limited to, trifluoromethoxy, trifluoroethoxy, monofluoromethoxy, monofluoroethoxy, difluoromethoxy, difluoroethoxy, and the like.

[0110] "Amino" refers to -NH2, "cyano" refers to -CN, "carboxy" refers to -C(O)OH, "acetyl" refers to -C(O)CH3, and "hydroxy" refers to -OH.

[0111] -Boc means tert-butyloxycarbonyl, -Tos means p-toluenesulfonyl, -Ts means p-toluenesulfonyl, -PMB means p-methoxybenzyl, -TBS means tert-butyldimethylsilyl, and -Ms means methanesulfonyl.

[0112] When the number of substituents is not indicated in the present application, it means that the substituents may be substituted with the number of substituents that can be substituted.

[0113] Unless otherwise defined, the "substituents independently selected from..." mentioned herein means that when more than one hydrogen on a group is replaced by a substituent, the substituents may be the same or different in type, and the substituents selected are independently of each other.

[0114] Unless otherwise defined, any group herein may be substituted or unsubstituted. When the above groups are substituted, the substituents are preferably 1 to 5 groups independently selected from deuterium, halogen (preferably fluorine, chlorine), cyano, hydroxyl, carboxyl, C 1-8 Alkyl (preferably C 1-6 Alkyl, more preferably C 1-3 Alkyl), C 1-8 Alkoxy (preferably C 1-6 Alkoxy, more preferably C 1-3 Alkoxy), C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-8 Alkyl (preferably halogenated C 1-6 Alkyl, more preferably halogenated C 1-3 Alkyl), cyano substituted C 1-8 Alkyl (preferably cyano substituted C 1-6 Alkyl, more preferably cyano substituted C 1-3 Alkyl), halogenated C 1-8 Alkoxy (preferably halogenated C 1-6 Alkoxy, more preferably halogenated C 1-3 Alkoxy), NR A0 R B0 、-SO2C 1-3 Alkyl, -S(O)C 1-3Alkyl, -C(O)NR A1 R B1 、-C(O)C 1-8 Alkyl (preferably -C(O)C 1-6 Alkyl, more preferably -C(O)C 1-3 Alkyl), -C(O)OC 1-8 Alkyl (preferably -C(O)OC 1-6 Alkyl, more preferably -C(O)OC 1-3 Alkyl), -OC(O)C 1-8 Alkyl (preferably -OC(O)C 1-6 Alkyl, more preferably -OC(O)C 1-3 Alkyl), C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl; wherein the 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl in the substituent is unsubstituted or substituted by 1, 2 or 3 groups independently selected from halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3 Alkoxy, NR A0 R B0 、-SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NR A1 R B1 、-C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl;

[0115] R A1 , R B1 are independently hydrogen or C 1-3 Alkyl; or R A1 , R B1 Together with the connected nitrogen atom, it forms a 4- to 6-membered saturated monocyclic heterocyclic ring; the 4- to 6-membered saturated monocyclic heterocyclic ring is unsubstituted or substituted by 1, 2 or 3 substituents independently selected from the following groups: deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3Alkoxy, -SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NH2, -C(O)NH(C 1-3 alkyl), -C(O)N(C 1-3 Alkyl)2, -C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl;

[0116] R A0 , R B0 are independently hydrogen, C 1-3 Alkyl or acetyl; or R A0 , R B0 Together with the connected nitrogen atom, it forms a 4- to 6-membered saturated monocyclic heterocyclic ring; the 4- to 6-membered saturated monocyclic heterocyclic ring is unsubstituted or substituted by 1, 2 or 3 substituents independently selected from the following groups: deuterium, halogen, cyano, hydroxyl, carboxyl, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, halogenated C 1-3 Alkoxy, -SO2C 1-3 Alkyl, -S(O)C 1-3 Alkyl, -C(O)NH2, -C(O)NH(C 1-3 alkyl), -C(O)N(C 1-3 Alkyl)2, -C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkyloxy, 3- to 6-membered heterocycloalkyl.

[0117] In the present application, when two or more “preferably” appear in one solution, any two “preferably” may be independent of each other.

[0118] In the present application, when the number of substituents is greater than 1, any two substituents may be the same or different. For example, they may be two same or different halogen substitutions, or they may be one halogen and one hydroxyl substitution.

[0119] Each type of substituent group described above may itself be substituted by the groups described herein.

[0120] Pharmaceutical composition

[0121] Generally, the compounds of the present invention or their pharmaceutically acceptable salts can be administered in a suitable dosage form with one or more pharmaceutical carriers. These dosage forms are suitable for oral administration, rectal administration, topical administration, oral administration, and other parenteral administration (e.g., subcutaneous, intramuscular, intravenous, etc.). For example, dosage forms suitable for oral administration include capsules, tablets, granules, and syrups.

[0122] "Pharmaceutically acceptable carrier" refers to a non-toxic, inert, solid, semi-solid substance or liquid filler, diluent, encapsulating material or auxiliary formulation or any type of excipient that is compatible with a patient, preferably a mammal, more preferably a human, and is suitable for delivering an active agent to a target site without terminating the activity of the agent.

[0123] The compositions of the present application are formulated, dosed and administered in a manner consistent with medical practice. The "therapeutically effective amount" of the compound administered is determined by factors such as the specific condition to be treated, the individual being treated, the cause of the condition, the target of the drug, and the mode of administration.

[0124] "Therapeutically effective amount" refers to the amount of the compound of the present invention that will induce a biological or medical response in an individual, such as reducing or inhibiting enzyme or protein activity or improving symptoms, alleviating symptoms, slowing or delaying disease progression, or preventing disease.

[0125] "Patient" refers to 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.

[0126] "Treatment" means to lessen, slow the progression, attenuate, prevent, or maintain an existing disease or condition (eg, cancer). Treatment also includes curing, preventing the development of, or alleviating to some extent, one or more symptoms of a disease or condition.

[0127] The "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0128] "Pharmaceutically acceptable acid addition salt" refers to a salt formed with an inorganic or organic acid that retains the biological effectiveness of the free base without other adverse effects.

[0129] "Pharmaceutically acceptable base addition salts" include, but are not limited to, salts with inorganic bases and salts with organic bases.

[0130] The compounds of the present application may contain one or more chiral centers and exist in different optically active forms. When the compound contains one chiral center, the compound comprises enantiomers. When the compound contains more than one chiral center, diastereomers may exist. The present application includes these two stereoisomers and mixtures of these two stereoisomers, such as racemates, diastereoisomer mixtures, etc.

[0131] The enantiomers, diastereomers and mixtures of these isomers of the compounds of the present application are all within the scope of protection of the present application. Enantiomers and diastereomers can be separated by methods known in the art, such as crystallization and chiral chromatography. Unless otherwise specified, the wedge-shaped bond is used. Represents the absolute configuration of a stereocenter. When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, unless otherwise specified, they include E and Z geometric isomers. Likewise, all tautomeric forms are included within the scope of this application.

[0132] Preparation method

[0133] The application provides a method for preparing a compound of formula (I), which can be synthesized using standard synthesis 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 provided herein can be changed according to the art. The reactions can be used in sequence to provide compounds of the application, or they can be used to synthesize fragments, which are subsequently added by methods described herein and / or methods known in the art.

[0134] The compounds described in this application can be synthesized using appropriate selectable starting materials using methods similar to the following or the exemplary methods described in the examples, or relevant public documents used by those skilled in the art. The starting materials or intermediates used to synthesize the compounds described in this application can be synthesized or can be obtained from commercial sources. If the existing literature is not reported or cannot be obtained from commercial sources, similar existing preparation methods of analogs or similar preparation methods recorded in this application can be used to prepare. Compounds such as described in this application and other related compounds with different substituents can be synthesized using techniques and raw materials known to those skilled in the art. The general method for preparing compounds disclosed in this application can be from reactions known in the art, and the reaction can be modified by reagents and conditions deemed appropriate by those skilled in the art to introduce various parts in the molecules provided by this application.

[0135] Compared with the prior art, the main advantages of the present application are: the compounds of the present application have novel structures and excellent SHP2 enzyme inhibitory activity, some of which are below 110nM, and some of which can even reach below 10nM; the compounds of the present application also have a significant degradation effect on SHP2 protein. The compounds of the present application also have excellent cytotoxicity, especially the cytotoxicity to MV411 cells is below μM, and some compounds can reach below 0.1μM; in short, the compounds of the present application have excellent SHP2 inhibitory activity and SHP2 protein degradation activity, and have significant cytotoxicity, and have potential therapeutic effects on diseases or conditions related to abnormal SHP2 activity.

[0136] The present application will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present application and are not intended to limit the scope of the present application. The experimental methods for which specific conditions are not indicated in the following examples are usually based on conventional conditions such as Sambrook et al., Molecular Cloning: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight. Unless otherwise defined, the terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any method and material similar or equivalent to the recorded content can all be applied to the present application.

[0137] Reagents and instruments

[0138] 1 H NMR: Bruker AVANCE-400 nuclear magnetometer, internal standard is tetramethylsilane (TMS).

[0139] LC-MS: Agilent 1290 HPLC System / 6130 / 6150MS liquid chromatography-mass spectrometer (manufacturer: Agilent), column: Waters BEH / CHS, 50×2.1 mm, 1.7 μm.

[0140] Preparative high performance liquid chromatography (pre-HPLC): GX-281 (Manufacturer: Gilson).

[0141] Use ISCO Combiflash-Rf75 or Rf200 automatic column analyzer and Agela 4g, 12g, 20g, 40g, 80g, 120g disposable silica gel columns.

[0142] Known starting materials can be used or synthesized according to methods known in the art, or can be purchased from companies such as ABCR MBH & Co. KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc, and Darui Chemicals.

[0143] In the embodiment, the reaction progress can be monitored by thin layer chromatography (TLC), and the compound purification can be performed by column chromatography. The developing solvent system used in column chromatography or TLC can be selected from: dichloromethane and methanol system, n-hexane and ethyl acetate system, petroleum ether and ethyl acetate system and acetone system, etc. The volume ratio of the solvent is adjusted according to the different polarities of the compound.

[0144] As used herein, DIEA: N,N-diisopropylethylamine, DMSO: dimethyl sulfoxide, MeOH: methanol, DCM: dichloromethane, HATU: 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, DMF: N,N-dimethylformamide, EA: ethyl acetate, PE: petroleum ether, PPh3: triphenylphosphine, THF: tetrahydrofuran, DIAD: diisopropyl azodicarboxylate, DEAD: diethyl azodicarboxylate, K2CO3: potassium carbonate, NMP: N-methylpyrrolidone, TFA: trifluoromethane Fluoroacetic acid, NaHCO3: sodium bicarbonate, KI: potassium iodide, NaI: sodium iodide, NaN3: sodium azide, T3P: 1-propylphosphoric acid cyclic anhydride, CuBr2: cupric bromide, H2O: water, PdCl2(PPh3)2: bis(triphenylphosphine)palladium(II) chloride, CuI: cuprous iodide, CuSO4: cupric sulfate, DDQ: 2,3-dichloro-5,6-dicyano-1,4-benzoquinone, NaBH4: sodium borohydride, NH4HCO3: ammonium bicarbonate, DTT: dithiothreitol, BSA: bovine serum albumin, FBS: fetal bovine serum.

[0145] As mentioned in this article, the percentage contents, unless otherwise specified, refer to mass percentage for solid-liquid mixing and solid-solid mixing, and refer to volume percentage for liquid-liquid mixing.

[0146] As used herein, room temperature refers to about 20-30°C.

[0147] As used herein, "overnight" means about 10 to 16 hours.

[0148] Preparation of intermediate Z1

[0149]

[0150] Step 1: Dissolve 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (0.552 g, 2 mmol), DIEA (0.78 g, 6 mmol), and 3-(2-(2-aminoethoxy)ethoxy)propionic acid (0.35 g, 2 mmol) in DMSO (6 mL). Stir at 100°C for 8 h. The reaction solution was concentrated and purified by flash column (12 g, 0-30% MeOH / DCM) to obtain Z1-a (450 mg, brown oil) with a yield of 52.1%. MS m / z (ESI): 434.1 [M+H] + .

[0151] Step 2: Dissolve Z1-a (0.433 g, 1 mmol), piperazine (0.13 g, 1.5 mmol), HATU (0.57 g, 1.5 mmol), and triethylamine (0.21 g, 2 mmol) in DMF (6 mL). Stir at room temperature for 0.5 h. After the reaction solution is concentrated, it is purified by flash column (12 g, 0-13% MeOH / DCM) to obtain intermediate Z1 (280 mg, brown oil) with a yield of 56.1%. MS m / z (ESI): 502.1 [M+H] + .

[0152] Preparation of intermediate Z2

[0153]

[0154] Step 1: Dissolve 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (0.552 g, 2 mmol), DIEA (0.78 g, 6 mmol), and 7-aminoheptan-1-ol (0.39 g, 3 mmol) in DMSO (6 mL). Stir at 100°C for 8 h. The reaction solution was concentrated and purified by flash column (12 g, 0-89% EA / PE) to obtain Z2-a (560 mg, brown oil) with a yield of 72.1%. MS m / z (ESI): 388.1 [M+H] + .

[0155] Step 2: Z2-a (0.338 g, 1 mmol), PPh3 (524 g, 2 mmol), carbon tetrabromide (0.664 g, 2 mmol) were dissolved in DCM (8 mL), stirred at room temperature for 2 h, and the reaction solution was concentrated and purified by flash column (12 g, 0-43% EA / PE) to obtain Z2-b (370 mg, brown oil), yield 82.2%, MS m / z (ESI): 450.1 [M+H] + .

[0156] Step 3: Dissolve Z2-b (0.225 g, 0.5 mmol), piperazine (0.07 g, 0.75 mmol), and DIEA (0.13 g, 1 mmol) in DMF (5 mL). Stir at 60°C for 3 h. Concentrate the reaction solution and purify it with a flash column (12 g, 0-10% MeOH / DCM) to obtain intermediate Z2 (210 mg, brown oil). Yield: 89.1%. MS m / z (ESI): 456.1 [M+H] + .

[0157] Preparation of intermediate Z3

[0158]

[0159] Step 1: 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (1.00 g, 3.65 mmol) was added to ultra-dry DMF (20 mL) at room temperature, and DIEA (0.9 mL, 5.47 mmol) was added, and the mixture was stirred for 5 minutes. Then, a solution of tert-butyl 2-bromoacetate (0.82 mL, 5.47 mmol) and ultra-dry DMF was added dropwise to the reaction solution, and then the temperature was raised to 30° C. for 4 hours. After adding water, a solid was precipitated. The solid was dried to obtain Z3-a (1.13 g, light blue solid, purity 95%), with a yield of 79.9%, MS m / z (ESI): 387.2 [MH] - .

[0160] Step 2: Add Z3-a (1.13 g, 2.90 mmol) to a hydrogen chloride / 1,4-dioxane solution (20 mL, 4 mol / L) at room temperature, and heat to 40°C for 2 hours. The reaction solution was concentrated and purified by slurrying with dichloromethane to obtain Z3-b (1.00 g, gray solid, crude product), MS m / z (ESI): 333.0 [M+H] + .

[0161] Step 3: Z3-b (500 mg, 1.51 mmol), tert-butyl piperazine-1-carboxylate (420.8 mg, 2.26 mmol), HATU (688.6 mg, 1.81 mmol), DIEA (0.75 mL, 4.53 mmol) were added to DMF (4 mL) at room temperature, and reacted at room temperature for 3 hours. The reaction solution was purified by column chromatography (EA) to obtain Z3-c (1.10 g, colorless oil, crude product), MS m / z (ESI): 499.3 [MH] - .

[0162] Step 4: Add Z3-c (1.1 g, 2.2 mmol) to a hydrogen chloride / 1,4-dioxane solution (10 mL, 4 mol / L) at room temperature. After the addition is complete, stir at room temperature overnight. The reaction solution is concentrated and dried under reduced pressure to obtain intermediate Z3 (1.00 g, yellow solid, crude product). MS m / z (ESI): 401.1 [M+H] + .

[0163] Preparation of intermediate Z4

[0164]

[0165] Step 1: Add tert-butyl 4-(2-hydroxyethyl)piperazine-1-carboxylate (1 g, 4.3 mmol) to ultra-dry THF (20 mL) at room temperature, then add PPh3 (1.13 g, 4.3 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (982 mg, 3.58 mmol). After addition, stir for 15 minutes, cool the system to 0°C, then drop DIAD (0.85 mL, 4.3 mmol) and ultra-dry THF (10 mL) solution, maintain the temperature at 0-10°C, add slowly to room temperature and react for 1 hour. After the reaction solution is concentrated, it is purified by column chromatography (MeOH / DCM=1 / 50) to obtain Z4-a (1.78 g, yellow solid, crude product), MS m / z (ESI): 485.2 [MH] - .

[0166] Step 2: Add Z4-a (780 mg, 1.605 mmol) to a hydrogen chloride / 1,4-dioxane solution (10 mL, 4 mol / L) at room temperature and stir overnight at room temperature. The reaction solution was concentrated to obtain intermediate Z4 (730 mg, white solid, crude product). MSm / z (ESI): 387.2 [M+H] + .

[0167] Preparation of intermediate Z5

[0168]

[0169] Step 1: 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (1.0 g, 3.65 mmol), 2-bromoethanol (684 mg, 5.47 mmol), and PPh3 (1.43 g, 5.47 mmol) were added to toluene (20 mL) at room temperature, and DEAD (952 mg, 5.47 mmol) was added dropwise under ice bath stirring. After the addition was completed, argon was replaced three times, and the temperature was slowly raised to 80 ° C for 6 hours. Water (60 mL) was added to the reaction solution, and ethyl acetate (100 mL × 3) was used for extraction. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE: EA = 1: 1) to obtain Z5-a (1.2 g, white solid, crude product). It was used directly in the next step without further treatment.

[0170] Step 2: Add Z5-a (1.08 g, 2.82 mmol) to DMF (10 mL) at room temperature, add tert-butyl 4-(piperidin-4-ylmethyl)piperazine-1-carboxylate (400 mg, 1.41 mmol) and K2CO3 (580 mg, 4.23 mmol) under stirring at room temperature, and after the addition is complete, heat to 70 ° C and react for 3 hours. Add water (50 mL) to the reaction solution, extract with ethyl acetate (100 mL), wash the obtained organic phase with saturated brine, dry with anhydrous sodium sulfate, concentrate and purify by column chromatography (PE: EA = 1: 1) to obtain Z5-b (360 mg, white solid, yield: 43.7%). 1 H NMR (400MHz, DMSO-d6) δ11.10(s,1H),7.82(d,J=8.4Hz,1H),7.46(d,J=1.6Hz,1H),7.37–7.34(m,1H),5.13–5.09(m,1H),4.28( s,2H),3.38–2.28(m,4H),2.99–2.51(m,7H),2.25(s,4H),2.12–1.90(m,5H),1.67–1.47(m,3H),1.38(s,9H),1.13–1.08(m,2H).

[0171] Step 3: Add Z5-b (230 mg, 0.394 mmol) to DCM (10 mL) at room temperature, and add hydrogen chloride / 1,4-dioxane solution (1 mL, 4 mol / L) under stirring at room temperature. After the addition is complete, stir at room temperature for 1 hour. Filter the reaction solution and dry the resulting filter cake to obtain intermediate Z5 (186 mg, white solid, crude product). Use it directly in the next step without further treatment.

[0172] Preparation of intermediate Z6

[0173]

[0174] Step 1: Add 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (1 g, 3.6 mmol) to ultra-dry THF (20 mL) at room temperature, then add PPh3 (1.15 g, 4.4 mmol), 2-(2-(2-chloroethoxy)ethoxy)ethan-1-ol (905 mg, 3.6 mmol), stir for 15 minutes, cool the system to 0-10°C, then add a solution of DIAD (890 mg, 4.4 mmol) and ultra-dry THF (10 mL) dropwise, maintain 0-10°C during the addition process, return to room temperature and react for 3 hours. The reaction solution was concentrated and purified by column chromatography (DCM / MeOH=50 / 1) to obtain Z6-a (3.00 g, yellow oil, crude product). MSm / z(ESI):424.9[M+H] + .

[0175] Step 2: Disperse Z6-a (1.0 g, 3.36 mmol) and tert-butyl piperazine-1-carboxylate (0.53 g, 4.40 mmol) in DMF (15 mL) at room temperature, add K2CO3 (0.98 g, 7.07 mmol) and potassium iodide (1.06 g, 7.07 mmol) under stirring at room temperature, and slowly heat the reaction solution to 80 ° C for 5 hours. Add water (50 mL) to the reaction solution, extract with ethyl acetate (100 mL × 3), combine the organic phase, wash with saturated brine (50 mL × 6), dry with anhydrous sodium sulfate, concentrate and purify by column chromatography (DCM / MeOH = 20 / 1) to obtain Z6-b (380 mg, yellow solid, purity 73%), yield: 28.0%. MSm / z (ESI): 575.3 [M+H] + .

[0176] Step 3: Add Z6-b (380 mg, 0.66 mmol) to hydrogen chloride / 1,4-dioxane solution (5 mL, 4 mol / L) at room temperature and react for 1 hour at room temperature. The reaction solution was concentrated to obtain intermediate Z6 (200 mg, yellow solid, purity 92.5%), with a yield of 63%, MS m / z (ESI): 475.3 [M+H] + .

[0177] Preparation of intermediate Z7

[0178]

[0179] Step 1: 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (1 g, 3.62 mmol) and tert-butyl 3-aminopropionate (683.36 mg, 4.71 mmol) were dissolved in NMP (15 mL), DIEA (1.40 g, 10.86 mmol) was added, and microwave heating was performed at 130°C for 50 minutes. After the reaction was cooled, water and ethyl acetate were added for dilution, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, dried over sodium sulfate, concentrated, and purified by silica gel column chromatography (PE:EA=4:1-1:4) to obtain Z7-a (360 mg, brown solid, purity 98%), yield: 24.8%, MS m / z (ESI): 346.1 [M-56+H] + .

[0180] Step 2: Dissolve Z7-a (360 mg, 896.83 μmol) in DCM (12 mL), then slowly add TFA (3 mL) dropwise, and react at room temperature for 4 hours. The reaction solution was concentrated to obtain Z7-b (350 mg, brown solid, crude product), MS m / z (ESI): 346.0 [M+H] + The crude product was directly used in the next reaction.

[0181] Step 3: Dissolve Z7-b (170 mg, 492.32 μmol) and tert-butyl piperazine-1-carboxylate (91.69 mg, 492.32 μmol) in THF (8 mL), add DIEA (254.51 mg, 1.97 mmol) and HATU (260.03 mg, 689.24 μmol), and react at room temperature for 1 hour. The reaction solution was concentrated and purified by silica gel column chromatography (DCM: MeOH = 100: 7) to obtain Z7-c (215 mg, brown solid, purity 97%), yield: 85.04%, MS m / z (ESI): 414.1 [M-100+H] + .

[0182] Step 4: Dissolve Z7-c (200 mg, 389.45 μmol) in DCM (8 mL), then slowly add TFA (2 mL) and react at room temperature for 4 hours. The reaction solution was concentrated and dissolved in DCM, then alkalized with triethylamine, concentrated and purified by silica gel column chromatography (DCM: MeOH = 10: 1) to obtain Z7 (100 mg, brown solid, purity 89%), yield: 62.1%, MS m / z (ESI): 414.1 [M+H] + .

[0183] Preparation of intermediate Z8

[0184]

[0185] Step 1: 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (1 g, 3.62 mmol), tert-butyl 3-(2-aminoethoxy)propionate (890.69 mg, 4.71 mmol) were dissolved in NMP (15 mL), DIEA (1.40 g, 10.86 mmol) was added, and then microwaved to 130°C for 50 minutes. After the reaction was cooled, water and ethyl acetate were added for dilution, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, dried over sodium sulfate, concentrated, and purified by silica gel column chromatography (PE:EA=4:1-1:4) to obtain Z8-a (920 mg, brown solid, purity 100%), yield 57.1%, MS m / z (ESI): 390.1 [M-56+H] + .

[0186] Step 2: Z8-a (850 mg, 1.91 mmol) was dissolved in DCM (15 mL), and then TFA (4 mL) was slowly added dropwise. The reaction mixture was reacted at room temperature for 4 hours. The reaction solution was concentrated to obtain Z8-b (700 mg, brown solid, crude product). MS m / z (ESI): 390.0 [M+H] + The crude product was directly used in the next reaction.

[0187] Step 3: Dissolve tert-butyl piperazine-1-carboxylate (95.67 mg, 513.67 μmol) and Z8-b (200 mg, 513.67 μmol) in THF (8 mL), add DIEA (265.55 mg, 2.05 mmol) and HATU (271.31 mg, 719.13 μmol), and react at room temperature for 1 hour. The reaction solution was concentrated and purified by silica gel column chromatography (DCM: MeOH = 100: 7) to obtain Z8-c (275 mg, brown solid, purity 95%), with a yield of 96.0%, MS m / z (ESI): 458.1 [M-100+H] + .

[0188] Step 4: Dissolve Z8-c (250 mg, 448.35 μmol) in DCM (8 mL), then slowly add TFA (2 mL) and react at room temperature for 4 hours. The reaction solution was concentrated and dissolved in dichloromethane, alkalized with triethylamine, and then concentrated and purified by silica gel column chromatography (DCM: MeOH = 10: 1) to obtain intermediate Z8 (180 mg, brown solid, purity 85%), with a yield of 87.7%, MS m / z (ESI): 458.0 [M+H] + .

[0189] Preparation of intermediate Z9

[0190]

[0191] Step 1: 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (1.5 g, 5.43 mmol) and tert-butyl N-(3-aminopropyl)carbamate (1.23 g, 7.06 mmol) were dissolved in NMP (15 mL), DIEA (2.11 g, 16.29 mmol) was added, and then microwaved to 130 degrees for reaction for 50 minutes. After the reaction was cooled, water and ethyl acetate were added for dilution, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (PE:EA=4:1-1:4) to obtain Z9-a (1.4 g, brown solid, purity 100%), with a yield of 59.9%, MS m / z (ESI): 331.1 [M-100+H] + .

[0192] Step 2: Dissolve Z9-a (1.4 g, 3.25 mmol) in DCM (20 mL), then slowly add TFA (5 mL) and react at room temperature for 4 h. The reaction solution was concentrated and purified by slurrying with methyl tert-butyl ether (150 mL) to obtain intermediate Z9 (912 mg, brown solid) with a yield of 84.9%, MS m / z (ESI): 331.2 [M+H] + .

[0193] Preparation of intermediate Z10

[0194]

[0195] Step 1: Dissolve 2-(2,6-dioxopiperidine-3-yl)-4-hydroxyisoindoline-1,3-dione (1 g, 3.65 mmol) and tert-butyl 3-[2-(2-bromoethoxy)ethoxy]propanoate (1.08 g, 3.65 mmol) in DMF (15 mL), add NaHCO3 (612.63 mg, 7.29 mmol), KI (60.53 mg, 364.66 μmol), heat to 70 ° C for 20 h, then add KI (60.53 mg, 364.66 μmol), NaHCO3 (612.63 mg, 7.29 mmol) and tert-butyl 3-[2-(2-bromoethoxy)ethoxy]propanoate (1.08 g, 3.65 mmol), and continue to react at 70 ° C for 10 h. The reaction solution was diluted with water and ethyl acetate, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (PE:EA=10:1-1:2) to obtain Z10-a (1.27 g, brown oil, purity 97.8%), yield: 71%, MS m / z (ESI): 435.2 [M-56+H] + .

[0196] Step 2: Dissolve Z10-a (1.27 g, 2.59 mmol) in DCM (15 mL), add TFA (3 mL), and stir at room temperature for 4 h. The reaction solution was concentrated to obtain Z10-b (1.27 g, brown oil, crude product), which was directly used in the next step. MS m / z (ESI): 435.0 [M+H] + .

[0197] Step 3: Dissolve Z10-b (1.2 g, 2.76 mmol) and tert-butyl piperazine-1-carboxylate (514.51 mg, 2.76 mmol) in THF (20 mL), add DIEA (1.43 g, 11.05 mmol) and HATU (1.46 g, 3.87 mmol), and stir at room temperature for 1 hour. The reaction solution was concentrated and purified by silica gel column chromatography (DCM: MeOH = 10: 1) to obtain Z10-c (1.2 g, brown solid, purity 94.3%), with a yield of 72.1%, MS m / z (ESI): 603.1 [M-100+H] + .

[0198] Step 4: Dissolve Z10-c (1.1 g, 1.83 mmol) in DCM (18 mL), add TFA (3.5 mL), and react at room temperature for 4 h. The reaction solution was concentrated and dissolved in dichloromethane, then triethylamine was added for neutralization, and concentrated and purified by silica gel column chromatography (DCM: MeOH = 8: 1) to obtain intermediate Z10 (700 mg, brown solid, purity 85.2%), yield 76.3%, MSm / z (ESI): 503.0 [M+H] + .

[0199] Preparation of intermediate Z11

[0200]

[0201] Referring to the preparation method of intermediate Z10, 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione was replaced with 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione to prepare intermediate Z11, MS m / z (ESI): 503.0 [M+H] + .

[0202] Preparation of intermediate Z12

[0203]

[0204] Step 1: Z12-a (200 mg, 443.00 μmol) was dissolved in DMF (3 mL), and NaN3 (34.56 mg, 531.60 μmol) was added, and the mixture was heated and stirred in an oil bath at 60°C for 2 h. Water (50 mL) was added to quench the mixture, and the mixture was extracted with ethyl acetate (20 mL×2). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, concentrated, and purified by Combi Flash silica gel column chromatography (4 g, 0-50% EA / PE, 0-5% MeOH / DCM) to obtain intermediate Z12 (251 mg, yellow solid), with a yield of 60.01%, MS m / z (ESI): 387.1 [M+H] + .

[0205] Preparation of intermediate Z13

[0206]

[0207] Step 1: Dissolve 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (1.4 g, 5.07 mmol) and tert-butyl 3-aminopropionate (1.03 g, 7.10 mmol) in NMP (12 mL), and add DIEA (1.97 g, 15.21 mmol). Heat the reaction to 110 ° C for 2 h in a microwave oven. Dilute the reaction solution with ethyl acetate (80 mL) and water (80 mL), extract the aqueous phase with ethyl acetate, and wash the combined organic phases with saturated sodium chloride, dry over anhydrous sodium sulfate, concentrate, and purify by silica gel column chromatography (PE: EA = 10: 1-1: 1) to obtain Z13-a (1.4 g, brown solid, purity 100%), with a yield of 68.81%, MS m / z (ESI): 346.0 [M-56+H] + .

[0208] Step 2: Dissolve Z13-a (1.4 g, 3.49 mmol) in DCM (40 mL), add TFA (11.91 g, 104.47 mmol), and stir at room temperature for 2 h. The reaction solution was concentrated and purified by slurrying with ethyl acetate to obtain Z13-b (0.7 g, brown solid, purity 96.3%), with a yield of 58.1%, MS m / z (ESI): 346.0 [M+H] + .

[0209] Step 3: Dissolve Z13-b (421.82 mg, 1.45 mmol) and tert-butyl 2-amino-7,7-difluoro-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (500 mg, 1.45 mmol) in THF (15 mL), add DIEA (374.28 mg, 2.90 mmol), and finally add T3P (2.80 g, 3.62 mmol, purity 50%), heat to 80 ° C in an oil bath and react for 2.5 h. After the reaction solution is concentrated, it is purified by silica gel column chromatography (PE:EA=1:1) to obtain Z13-c (650 mg, brown solid, purity 98.8%), yield: 72.6%, MS m / z (ESI): 619.3 [M+H] + .

[0210] Step 4: Dissolve Z13-c (650 mg, 1.05 mmol) in DCM (20 mL), add TFA (3 mL), and react at room temperature for 2 h. The reaction solution was concentrated and dissolved in DCM, then alkalized with triethylamine, concentrated and purified by silica gel column chromatography (DCM: MeOH = 100: 1-100: 6) to obtain intermediate Z13 (350 mg, brown oil, purity 88.4%), yield 64.24%, MS m / z (ESI): 518.9 [M+H] + .

[0211] Preparation of intermediate Z14

[0212]

[0213] Step 1: Dissolve tert-butyl 2-amino-7,7-difluoro-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (0.582 g, 2 mmol), CuBr2 (0.57 g, 3 mmol), and isoamyl nitrite (0.39 g, 3 mmol) in acetonitrile (10 mL), and stir at 65 ° C for 1 h. The reaction solution was concentrated and purified by flash column (12 g, 0-56% EA / PE) to obtain Z14-a (560 mg, brown oil) with a yield of 78.8%. MS m / z (ESI): 354.9; 356.9 [M+H] + .

[0214] Step 2: 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (0.357 g, 1 mmol), DIEA (0.39 g, 3 mmol), tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (0.2 g, 1 mmol) were dissolved in DMSO (5 mL), stirred at 100 ° C for 8 h, and the reaction solution was concentrated and purified by flash column (12 g, 0-89% EA / PE) to obtain Z14-b (380 mg, brown oil) with a yield of 84.4%. MS m / z (ESI): 455.1 [M+H] + .

[0215] Step 3: Z14-b (278 mg, 0.5 mmol) was dissolved in DCM (3 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature for 0.5 h. The reaction was completed by LCMS. The reaction solution was concentrated to give Z14-c (200 mg, brown oil) with a yield of 100%, which was used directly in the next step. MS m / z (ESI): 355.1 [M+H] + .

[0216] Step 4: Z14-c (0.20 g, 0.56 mmol), DIEA (0.20 g, 1.6 mmol), and compound Z14-a (0.2 g, 0.56 mmol) were dissolved in DMSO (5 mL), stirred at 100 ° C for 8 h, and the reaction solution was concentrated and purified by flash column (12 g, 0-80% EA / PE) to obtain Z14-d (120 mg, brown oil) with a yield of 32.1%. MS m / z (ESI): 629.1 [M+H] + .

[0217] Step 5: Dissolve Z14-d (0.063 g, 0.1 mmol) in MeOH (3 mL), add hydrogen chloride / 1,4-dioxane (0.5 mL, 4 mol / L), and stir at room temperature for 0.2 h. The reaction solution was concentrated to obtain intermediate Z14, which was used directly in the next step. MS m / z (ESI): 529.1 [M+H] + .

[0218] Preparation of intermediate Z15

[0219]

[0220] Step 1: Dissolve 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (1.1 g, 4 mmol) and methyl 4-bromobutyrate (0.73 g, 4 mmol) in DMF (10 mL), add NaI (0.60 g, 4 mmol) and NaHCO3 (0.67 g, 8 mmol), stir at 70°C for 24 h, concentrate the reaction solution and purify it with a flash column (12 g, 0-95% EA / PE) to obtain Z15-a (1.5 g, brown solid), with a yield of 99.9%. MS m / z (ESI): 375.1 [M+H] + .

[0221] Step 2: Dissolve Z15-a (1.5 g, 4 mmol) in water (30 mL), add TFA (13 mL), stir at 70 ° C for 3 h, and the reaction is complete after LCMS detection. The reaction solution is concentrated and purified by flash column (20 g, 0-25% MeOH / DCM) to obtain Z15-b (1 g, white solid) with a yield of 71.4%, MS m / z (ESI): 361.1 [M+H] + .

[0222] Step 3: Z15-b (0.360 g, 1 mmol), tert-butyl 2-amino-7,7-difluoro-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (0.291 g, 1 mmol), T3P (0.99 g, 3 mmol), DIEA (0.26 g, 2 mmol) were dissolved in THF (8 mL) and stirred at room temperature for 0.5 h. The reaction solution was concentrated and purified by flash column (12 g, 0-70% EA / PE) to obtain Z15-c (390 mg, brown oil) with a yield of 61.9%. MS m / z (ESI): 634.1 [M+H] + .

[0223] Step 4: Dissolve Z15-c (0.312 g, 0.5 mmol) in MeOH (8 mL), add hydrogen chloride / 1,4-dioxane solution (1.0 mL), stir at room temperature for 0.2 h, and concentrate the reaction solution to obtain intermediate Z15, which is used directly in the next step. MS m / z (ESI): 534.1 [M+H] + .

[0224] Preparation of intermediate Z16

[0225]

[0226] Step 1: Z14-a (0.291 g, 1 mmol), DIEA (0.39 g, 3 mmol), butane-1,4-diamine (0.1 g, 1.2 mmol) were dissolved in DMSO (5 mL), stirred at 100 ° C for 8 h, and the reaction solution was concentrated and purified by flash column (12 g, 0-98% EA / PE) to obtain Z16-a (343 mg, brown oil) with a yield of 99.8%. MS m / z (ESI): 363.1 [M+H] + .

[0227] Step 2: Z16-a (0.343 g, 1 mmol), DIEA (0.39 g, 3 mmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.274 g, 1 mmol) were dissolved in DMSO (5 mL), stirred at 100 ° C for 8 h, and the reaction solution was concentrated and purified by flash column (12 g, 0-78% EA / PE) to obtain Z16-b (210 mg, brown oil) with a yield of 34.4%. MS m / z (ESI): 619.1 [M+H] + .

[0228] Step 3: Dissolve Z16-b (0.210 g, 0.34 mmol) in DCM (8 mL), add TFA (2 mL), stir at room temperature for 0.5 h, and concentrate the reaction solution to obtain intermediate Z16, which is used directly in the next step. MS m / z (ESI): 519.1 [M+H] + .

[0229] Preparation of intermediate Z17

[0230]

[0231] Step 1: Dissolve 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (0.274 g, 1 mmol) and tert-butyl 3-hydroxypropylcarbamate (0.175 g, 1 mmol) in THF (8 mL), add PPh3 (0.39 g, 1.5 mmol) and DIAD (0.3 g, 1.5 mmol), stir at room temperature for 1 h, concentrate the reaction solution and purify it with a flash column (12 g, 0-70% EA / PE) to obtain Z17-a (360 mg, brown oil), yield 83.7%, MS m / z (ESI): 432.1 [M+H] + .

[0232] Step 2: Z17-a (360 mg, 0.84 mmol) was dissolved in DCM (8 mL), TFA (2 mL) was added, and the mixture was stirred at room temperature for 0.5 h. The reaction was complete when detected by LCMS. The reaction solution was concentrated to give Z17-b (300 mg, brown oil) with a yield of 100%, which was used directly in the next step. MS m / z (ESI): 332.1 [M+H] + .

[0233] Step 3: Z17-b (0.3 g, 0.91 mmol), DIEA (0.20 g, 2.7 mmol), and Z14-a (0.34 g, 0.91 mmol) were dissolved in DMSO (7 mL), stirred at 100 ° C for 8 h, and the reaction solution was concentrated and purified by flash column (12 g, 0-80% EA / PE) to obtain Z17-c (120 mg, brown oil) with a yield of 22.2%. MS m / z (ESI): 606.1 [M+H] + .

[0234] Step 4: Dissolve Z17-c (0.12 g, 0.2 mmol) in DCM (5 mL), add TFA (1.5 mL), stir at room temperature for 0.5 h, and concentrate the reaction solution to obtain intermediate Z17, which is directly used in the next step. MS m / z (ESI): 506.1 [M+H] + .

[0235] Preparation of intermediate Z18

[0236]

[0237] Referring to the preparation method of intermediate Z17, (3-hydroxypropyl) tert-butyl carbamate was replaced with (4-hydroxybutyl) tert-butyl carbamate to prepare intermediate Z18, MS m / z (ESI): 520.1 [M+H] + .

[0238] Preparation of intermediate Z19

[0239]

[0240] Step 1: Dissolve tert-butyl 2-amino-7,7-difluoro-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (1.5 g, 5.15 mmol), hex-5-ynoic acid (866 mg, 7.72 mmol), and DIEA (3.99 g, 30.89 mmol) in THF (30 mL), add T3P (4.91 g, 7.72 mmol, 50% ethyl acetate solution), and stir the reaction solution at 80°C for 12 h. The reaction solution was concentrated and purified by column chromatography (PE:EA=3:1) to obtain Z19-a (1.42 g, yellow solid), with a yield of 71%, MS m / z (ESI): 386.2 [M+H] + .

[0241] Step 2: Dissolve Z19-a (200 mg, 0.519 mmol), 4-bromo-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (168 mg, 0.519 mmol), PdCl2(PPh3)2 (36 mmol, 51 μmol), and CuI (20 mg, 104 μmol) in triethylamine (3 mL) and DMF (6 mL) solution. Stir the reaction solution at 90 ° C for 12 h under nitrogen protection. After the reaction solution is concentrated, it is separated and purified by column chromatography (PE: EA = 2: 1) to obtain Z19-b (310 mg, yellow solid) with a yield of 95%. MS m / z (ESI): 528.2 [M+H-100] + ,572.2[M+H-56] + .

[0242] Step 3: Dissolve Z19-b (300 mg, 0.478 mmol) in TFA (3 mL) and stir at room temperature for 1 h. The reaction solution was concentrated and purified by column chromatography (PE: EA = 1: 1) to obtain intermediate Z19 (210 mg, white solid) with a yield of 83%. MSm / z (ESI): 528.2 [M+H] + .

[0243] Preparation of intermediate Z20

[0244]

[0245] Step 1: 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (2.0 g, 7.29 mmol), tert-butyl 5-bromopentanoate (1.9 g, 8.02 mmol), NaHCO3 (1.23 g, 14.59 mmol) and KI (1.21 g, 8.08 mmol) were dissolved in DMF (30 mL), and the reaction solution was stirred at 100°C for 10 h. The reaction solution was concentrated and purified by column chromatography (DCM: MeOH = 20: 1) to obtain Z20-a (1.46 g, yellow oil) with a yield of 46%, MS m / z (ESI): 375.1 [M+H-56] + .

[0246] Step 2: Dissolve Z20-a (200 mg, 0.519 mmol) in TFA (5 mL) and DCM (15 mL) solution, and stir the reaction solution at room temperature for 6 h. After the reaction solution is concentrated, it is separated and purified by column chromatography (DCM: MeOH = 15: 1) to obtain Z20-b (1.2 g, yellow oil), with a yield of 94%, MS m / z (ESI): 375.1 [M+H] + .

[0247] Step 3: Z20-b (1.2 g, 3.19 mmol), tert-butyl 2-amino-7,7-difluoro-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (0.93 g, 3.19 mmol), and DIEA (4.13 g, 31.92 mmol) were dissolved in THF (25 mL) and stirred at 60°C for 2 h. The reaction solution was concentrated and purified by preparative liquid chromatography (preparative column: Waters-SunFire C18, elution phase: 0.05% NH4HCO3 aqueous solution / acetonitrile, elution gradient: start 20% acetonitrile-end 80% acetonitrile, flow rate: 30 mL / min) to obtain Z20-c (730 mg, yellow solid), yield: 35%. MS m / z (ESI): 648.3 [M+H] + .

[0248] Step 4: Dissolve Z20-c (730 mg, 1.13 mmol) in TFA (2 mL) and DCM (10 mL) solution, and stir the reaction solution at room temperature for 2 h. After the reaction solution is concentrated, it is separated and purified by column chromatography (DCM: MeOH = 10: 1) to obtain intermediate Z20 (605 mg, white solid), yield: 98%. MS m / z (ESI): 548.2 [M+H] + .

[0249] Preparation of intermediate Z21

[0250]

[0251] Step 1: Z14-a (1.46 g, 4.10 mmol) and 3-aminopropan-1-ol (616.61 mg, 8.21 mmol) were dissolved in DMSO (10 mL), and DIEA (1.59 g, 12.31 mmol, 2.14 mL) was added, and the reaction was carried out under microwave at 100°C for 1 h. The reaction solution was poured into a separatory funnel, and water (150 mL) was added, and ethyl acetate was extracted (2×30 mL). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column (20 g, 0-4% DCM / MeOH) to obtain Z21-a (1.08 g, light yellow solid); yield: 75.3%; MS m / z (ESI): 350.1 [M+H] + .

[0252] Step 2: Dissolve Z21-a (500 mg, 1.43 mmol) and 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindole-1,3-dione (274.70 mg, 1.00 mmol) in THF (15 mL), add PPh3 (563.01 mg, 2.15 mmol), and then slowly dropwise add DIAD (578.14 mg, 2.86 mmol) dissolved in THF (3 mL), and react at room temperature for 12 hours. After the reaction solution is concentrated, it is purified by silica gel column chromatography (DCM: MeOH = 100: 5) to obtain Z21-b (350 mg, crude product). MS m / z (ESI): 606.0 [M+H] + .

[0253] Step 3: Dissolve Z21-b (150 mg, 242.08 μmol) in DCM (7 mL) and TFA (1.5 mL) and react at room temperature for 2 h. The reaction solution was concentrated and purified by silica gel column chromatography (DCM: MeOH = 100: 5) to obtain intermediate Z21 (100 mg, brown solid, crude product), MS m / z (ESI): 505.9 [M+H] + .

[0254] Preparation of intermediate Z22

[0255]

[0256] Step 1: Dissolve 2-(2,6-dioxopiperidine-3-yl)-4-hydroxyisoindoline-1,3-dione (582.11 mg, 2.12 mmol) and ethyl 2-[2-[2-[2-(tert-butoxycarbonylamino)ethoxy]ethoxy]-ethoxy]4-methylbenzenesulfonate (950 mg, 2.12 mmol) in DMF (8 mL), add KI (140.95 mg, 849.08 μmol) and K2CO3 (586.74 mg, 4.25 mmol), and heat the reaction to 40°C for 6 h. The reaction solution was diluted with water and ethyl acetate, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (DCM; MeOH = 10:1) to obtain Z22-a (650 mg, brown solid, purity 81%), with a yield of 55.7%, MS m / z (ESI): 450.1 [M-100+H] + .

[0257] Step 2: Dissolve Z22-a (650 mg, 1.18 mmol) in DCM (15 mL), add TFA (4 mL), and react at room temperature for 4 h. The reaction solution was concentrated to obtain Z22-b (800 mg, crude product), MS m / z (ESI): 450.0 [M+H] + .

[0258] Step 3: Z22-b (506.14 mg, 1.13 mmol) and Z14-a (200 mg, 563.07 μmol) were dissolved in DMSO (7 mL), DIEA (2.91 g, 22.52 mmol) was added, and the mixture was reacted at 150 °C for 2.5 h in a microwave oven. Ethyl acetate was added to dilute the reaction solution, the aqueous phase was extracted twice with ethyl acetate, the combined organic phases were washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (DCM: MeOH = 10: 1) to obtain Z22-c (85 mg, brown solid, purity 71%), with a yield of 14.6%, MS m / z (ESI): 724.3 [M+H] + .

[0259] Step 4: Dissolve Z22-c (85 mg, 117.45 μmol) in DCM (4 mL), add TFA (1 mL), and stir at room temperature for 3 h. The reaction solution was concentrated and dissolved in DCM, alkalized with triethylamine, concentrated, and purified by silica gel column chromatography (DCM: MeOH = 10: 1) to obtain intermediate Z22 (70 mg, brown solid, purity 90.8%), yield 95.6%, MS m / z (ESI): 624.3 [M+H] + .

[0260] Preparation of intermediate Z23

[0261]

[0262] Referring to the preparation method of intermediate Z16, butane-1,4-diamine was replaced with propane-1,3-diamine, and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione was replaced with 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione to prepare intermediate Z23, MS m / z (ESI): 505.2 [M+H] + .

[0263] Preparation of intermediate Z24

[0264]

[0265] Step 1: Propyl-2-yn-1-amine (395.41 mg, 7.18 mmol) and Z14-a (1.7 g, 4.79 mmol) were dissolved in DMSO (8 mL), and DIEA (1.86 g, 14.36 mmol, 2.50 mL) was added, and the mixture was stirred and reacted at 130°C for 4 h under microwave heating. Water (150 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (40 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by CombiFlash silica gel column (24 g, 0-17% EA / PE) to obtain Z24-a (291 mg, white solid, purity 67%), with a yield of 12.37%, MS m / z (ESI): 330.0 [M+H] + .

[0266] Step 2: Trimethylsilyl azide (50.97 mg, 442.44 μmol) and Z24-a (145 mg, 294.9 μmol) were dissolved in THF (1.5 mL) and H2O (1.5 mL), and sodium ascorbate (5.84 mg, 29.50 μmol) and CuSO4 (9.42 mg, 58.99 μmol) were added, and the reaction was stirred at 20°C for 14 h. Water (10 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (40 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by Combi Flash silica gel column (4 g, 0-20% EA / PE, 0-5% MeOH / DCM) to obtain Z24-b (66 mg, dark green solid), with a yield of 60.09%, MS m / z (ESI): 373.1 [M+H] + .

[0267] Step 3: Dissolve Z12-a (89.98 mg, 174.55 μmol) and Z24-b (65 mg, 174.55 μmol) in DMF (2 mL), add K2CO3 (72.37 mg, 523.64 μmol), and heat and stir in an oil bath at 65°C for 2 h. Concentrate the reaction solution and purify it with a Combi Flash silica gel column (4 g, 0-3% MeOH / DCM) to obtain Z24-c (64 mg, yellow solid, purity 67%), with a yield of 34.32%, MS m / z (ESI): 716.3 [M+H] + .

[0268] Step 4: Dissolve Z24-c (71 mg, 99.20 μmol) in DCM (2.0 mL), add TFA (113.11 mg, 992.00 μmol), and stir at 20°C for 40 min. Add DCM (40 mL) to the reaction solution, concentrate, then add DCM (40 mL) and an appropriate amount of triethylamine to obtain a clear solution, and concentrate to obtain intermediate Z24 (85 mg, yellow viscous solid, purity 67%), yield 93.26%, MS m / z (ESI): 616.2 [M+H] + .

[0269] Preparation of intermediate Z25

[0270]

[0271] Step 1: Disperse 5-fluoro-1-oxo-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-carboxylic acid tert-butyl ester (10 g, 31.3 mmol) and (R)-(+)-tert-butylsulfenamide (3.8 g, 31.3 mmol) in tetraethyl titanate (150 mL) at room temperature. After addition, the reaction solution was reacted at 120°C for 12 h. Extracted with ethyl acetate (100 mL×3). The organic phases were combined, washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE / EA=5 / 1) to obtain Z25-a (8.1 g, white solid, purity 73%), with a yield of 61%, MS m / z (ESI): 423.0[M+H] + .

[0272] Step 2: Disperse Z25-a (1.6 g, 1.57 mmol) and p-methoxybenzylamine (519 mg, 3.80 mmol) in DMSO (25 mL) at room temperature, add DIEA (1.40 g, 11.36 mmol) in batches, and react at 105°C for 4 h after the addition is complete. The reaction solution is extracted with ethyl acetate (100 mL × 3), the organic phases are combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE / EA = 5 / 1) to obtain Z25-b (850 mg, white solid, purity 76%), with a yield of 42%, MS m / z (ESI): 540.0 [M+H] + .

[0273] Step 3: Disperse Z25-b (850 mg, 3.80 mmol) in THF (25 mL) at room temperature, add DDQ (358 mg, 1.57 mmol) in batches, and react the reaction solution at room temperature for 4 h. The reaction solution was extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE / EA = 5 / 1) to obtain Z25-c (350 mg, brown solid, purity 80%), with a yield of 53%, MS m / z (ESI): 420.0 [M+H] + .

[0274] Step 4: Disperse Z25-c (350 mg, 0.83 mmol) in THF (25 mL) at room temperature, add diisobutylaluminum hydride (1.66 ml, 1.66 mmol) in batches, and react the reaction solution at 25 ° C for 4 h. The reaction solution was extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH = 20 / 1) to obtain Z25-d (280 mg, yellow solid, purity 89%), yield: 80%, MS m / z (ESI): 422.0 [M+H] + .

[0275] Step 5: Disperse Z25-d (280 mg, 0.67 mmol), monomethyl succinate (88 mg, 0.67 mmol), and triethylamine (68 mg, 0.67 mmol) in DMF (10 mL) at room temperature, add HATU (254 mg, 0.67 mmol) in batches, and react the reaction solution at room temperature for 4 h. The reaction solution was extracted with ethyl acetate (100 mL × 3), the organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE / EA = 3 / 1) to obtain Z25-e (285 mg, yellow solid, purity 83%), with a yield of 80%, MS m / z (ESI): 536.0 [M+H] + .

[0276] Step 6: Disperse Z25-e (285 mg, 0.53 mmol) in THF (10 mL) and water (2 mL) at room temperature, add lithium hydroxide (25 mg, 1.06 mmol) in batches, and react the reaction solution at room temperature for 4 h. The reaction solution was extracted with ethyl acetate (100 mL × 3), the organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH = 15 / 1) to obtain Z25-f (240 mg, yellow solid, purity 91%), with a yield of 86%, MS m / z (ESI): 522.0 [M+H] + .

[0277] Step 7: Disperse Z25-f (70 mg, 0.13 mmol), intermediate Z9 (44 mg, 0.13 mmol), and triethylamine (13 mg, 0.13 mmol) in DMF (5 mL) at room temperature, add HATU (49 mg, 0.13 mmol) in batches, and react the reaction solution at room temperature for 4 h. The reaction solution was extracted with ethyl acetate (100 mL × 3), the organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH = 15 / 1) to obtain intermediate Z25 (78 mg, yellow solid, purity 95%), yield: 69%, MS m / z (ESI): 834.0 [M+H] + .

[0278] Preparation of intermediate Z26

[0279]

[0280] Step 1: Disperse intermediate Z7 (60 mg, 0.15 mmol), Z25-f (75 mg, 0.15 mmol), and triethylamine (15 mg, 0.15 mmol) in DMF (5 mL) at room temperature, add HATU (57 mg, 0.15 mmol) in batches under stirring at room temperature, and react the reaction solution at room temperature for 2 h. The reaction solution was extracted with ethyl acetate (100 mL × 3), the organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH = 20 / 1) to obtain intermediate Z26 (61 mg, white solid, purity 90%), yield: 46%, MS m / z (ESI): 917.0 [M+H] + .

[0281] Preparation of intermediate Z27

[0282]

[0283] Step 1: Disperse 5-fluoro-1-oxo-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-carboxylic acid tert-butyl ester (2.0 g, 6.26 mmol) and 2-(2-((tert-butyldimethylsilyl)oxy)ethoxy)ethan-1-ol (74 mg, 0.27 mmol) in DMF (25 mL) at room temperature, add potassium tert-butoxide (2.1 g, 18.79 mmol) in batches under stirring at room temperature, and react the reaction solution at 60°C for 1 h. The reaction solution was extracted with ethyl acetate (100 mL×3), the organic phases were combined, washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE / EA=5 / 1) to obtain Z27-a (810 mg, white solid, purity 88%), with a yield of 25%, MS m / z (ESI): 464.1 [M-56+H] + .

[0284] Step 2: Disperse Z27-a (810 mg, 1.75 mmol) and (R)-(+)-tert-butylsulfenamide (283 mg, 2.34 mmol) in tetraethyl titanate (25 mL) at room temperature, and react the reaction solution at 120°C for 12 h. The reaction solution was extracted with ethyl acetate (100 mL×3). The organic phases were combined, washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE / EA=5 / 1) to obtain Z27-b (520 mg, white solid, purity 78%), with a yield of 54%, MS m / z (ESI): 623.0 [M+H] + .

[0285] Step 3: Disperse Z27-b (520 mg, 0.84 mmol) in THF (25 mL) at room temperature, add diisobutyl aluminum hydride (1.68 ml, 1.68 mmol) in batches under stirring at room temperature, and after the addition is completed, the reaction solution is reacted at -78 ° C for 1 h, and then the reaction solution is kept at 0 ° C for 1 h. The reaction solution is extracted with ethyl acetate (100 mL × 3), the organic phases are combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE / EA = 5 / 1) to obtain Z27-c (410 mg, white solid, purity 88%), with a yield of 79%, MS m / z (ESI): 625.0 [M+H] + .

[0286] Step 4: Disperse Z27-c (300 mg, 0.48 mmol) in THF (25 mL) at room temperature, add tetrabutylammonium bromide (251 mg, 0.96 mmol) in batches under stirring at room temperature, and after the addition is completed, the reaction solution is reacted at room temperature for 2 h. The reaction solution is extracted with ethyl acetate (100 mL × 3), the organic phases are combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE / EA = 5 / 1) to obtain Z27-d (180 mg, white solid, purity 91%), with a yield of 78%, MS m / z (ESI): 511.0 [M+H] + .

[0287] Step 5: Disperse Z27-d (180 mg, 0.35 mmol) and 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (97 mg, 0.35 mmol) in THF (25 mL) at room temperature, add PPh3 (92 mg, 0.35 mmol) and DIAD (70 mg, 0.35 mmol) in batches under stirring at room temperature, and react the reaction solution at 25°C for 2 h. The reaction solution was extracted with ethyl acetate (100 mL×3), the organic phases were combined, washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH=20 / 1) to obtain intermediate Z27 (120 mg, white solid, purity 75%), with a yield of 45%, MS m / z (ESI): 767.0 [M+H] + .

[0288] Preparation of intermediate Z28

[0289]

[0290] Step 1: Z25-f (200 mg, 0.383 mmol), 5-(3-aminopropoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione hydrochloride (169 mg, 0.460 mmol), and HATU (160 mg, 0.42 mmol) were added to a mixture of DMF and THF (1 mL / 1 mL) at room temperature, and triethylamine (0.11 mL, 0.766 mmol) was added dropwise under stirring at room temperature. The reaction solution was reacted at room temperature for 3 hours. The reaction solution was concentrated and purified by column chromatography (DCM / MeOH=20 / 1) to obtain intermediate Z28 (260 mg, transparent oil, crude product), MS m / z (ESI): 833.1 [MH] - .

[0291] Preparation of intermediate Z29

[0292]

[0293] Step 1: Disperse 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (1.0 g, 3.65 mmol) and oxybis(ethane-2,1-diyl)bis(4-methylbenzenesulfonate) (1.51 g, 3.65 mmol) in DMSO (15 mL) at room temperature, add K2CO3 (1.5 g, 10.95 mmol) in batches under stirring at room temperature, and react the reaction solution at 100°C for 2 h. The reaction solution was extracted with ethyl acetate (100 mL×3), the organic phases were combined, washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH=20 / 1) to obtain Z29-a (560 mg, white solid, purity 78%), with a yield of 30%, MS m / z (ESI): 517.0 [M+H] + .

[0294] Step 2: Disperse Z29-a (560 mg, 1.1 mmol), tert-butyl piperazine-1-carboxylate (205 mg, 1.1 mmol), and triethylamine (111 mg, 1.1 mmol) in DMSO (5 mL) at room temperature, add DIEA (143 mg, 1.1 mmol) in batches under stirring at room temperature, and react the reaction solution at 100 ° C for 3 h. The reaction solution was extracted with ethyl acetate (100 mL × 3), and the organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH = 20 / 1) to obtain Z29-b (270 mg, white solid, purity 92%), with a yield of 53%, MS m / z (ESI): 531.0 [M+H] + .

[0295] Step 3: Disperse Z29-b (270 mg, 0.13 mmol) in hydrogen chloride / 1,4-dioxane (5 mL, 4 mol / L) at room temperature, and react the reaction solution at room temperature for 2 h. The reaction solution was extracted with ethyl acetate (100 mL × 3), and the organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH = 20 / 1) to obtain Z29-c (121 mg, yellow solid, purity 65%), with a yield of 55%, MS m / z (ESI): 431.0 [M+H] + .

[0296] Step 4: Disperse Z29-c (121 mg, 0.28 mmol), Z25-f (140 mg, 0.28 mmol), and triethylamine (28 mg, 0.28 mmol) in DMF (5 mL) at room temperature, add HATU (106 mg, 0.28 mmol) in batches under stirring at room temperature, and react the reaction solution at 25°C for 2 h. The reaction solution was extracted with ethyl acetate (100 mL × 3), the organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH = 20 / 1) to obtain intermediate Z29 (180 mg, white solid, purity 81%), with a yield of 69%, MS m / z (ESI): 934.0 [M+H] + .

[0297] Preparation of intermediate Z30

[0298]

[0299] Step 1: Disperse (R,Z)-1-((tert-butylsulfinyl)imino)-5-fluoro-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-carboxylic acid tert-butyl ester (500 mg, 1.18 mmol) and propargylamine (65 mg, 1.18 mmol) in DMSO (5 mL) at room temperature, add DIEA (460 mg, 3.54 mmol) in batches under stirring at room temperature, and react at 90°C for 12 h. The reaction solution was extracted with ethyl acetate (100 mL×3), the organic phases were combined, washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH=20 / 1) to obtain Z30-a (310 mg, brown solid, purity 85%), yield: 57%, MSm / z (ESI): 458.0 [M+H] + .

[0300] Step 2: Disperse Z30-a (310 mg, 0.68 mmol) in THF (25 mL) at room temperature, add diisobutylaluminum hydride (1.36 mL, 1.36 mmol) in batches, and after the addition is complete, react the reaction solution at -78°C for 4.0 hours. The reaction solution is extracted with ethyl acetate (100 mL×3). The organic phases are combined, washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH=20 / 1) to obtain Z30-b (240 mg, yellow solid, purity 81%), with a yield of 77%, MS m / z (ESI): 460.0 [M+H] + .

[0301] Step 3: Disperse 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (500 mg, 1.82 mmol) and ((oxybis(ethane-2,1-diyl))bis(oxy))bis(ethane-2,1-diyl)bis(4-methylbenzenesulfonate) (916 mg, 1.82 mmol) in DMSO (10 mL) at room temperature. Add DIEA (707 mg, 5.47 mmol) in batches under stirring at room temperature. The reaction solution is reacted at 110°C for 2.0 hours. The reaction solution was extracted with ethyl acetate (100 mL × 3), and the organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH = 20 / 1) to obtain Z30-c (80 mg, white solid, purity 75%), with a yield of 7%, MS m / z (ESI): 605.0 [M+H] + .

[0302] Step 4: Disperse Z30-c (80 mg, 0.13 mmol) in DMF (5 mL) at room temperature, add sodium azide (8.5 mg, 0.13 mmol) in batches under stirring at room temperature, and react at 60°C for 2.0 hours. The reaction solution was extracted with ethyl acetate (100 mL × 3), and the organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH = 20 / 1) to obtain Z30-d (30 mg, white solid, purity 62%), with a yield of 51%, MS m / z (ESI): 448.0 [M-28+H] + .

[0303] Step 5: Disperse Z30-d (30 mg, 0.07 mmol) and Z30-b (32 mg, 0.07 mmol) in THF (5 mL) and water (1 ml) at room temperature, add sodium ascorbate (3 mg, 0.014 mmol) and copper sulfate (2.5 mg, 0.014 mmol) in batches under stirring at room temperature, and react the reaction solution at 25°C for 12 h. The reaction solution was extracted with ethyl acetate (100 mL × 3), the organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH = 20 / 1) to obtain intermediate Z30 (27 mg, white solid, purity 62%), with a yield of 41%, MS m / z (ESI): 957.0 [M+Na] + .

[0304] Preparation of intermediate Z31

[0305]

[0306] Step 1: Referring to the preparation method of intermediate Z10, 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione was replaced with 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione, and tert-butyl 3-[2-(2-bromoethoxy)ethoxy]propionate was replaced with tert-butyl 3-(2-hydroxyethoxy)propionate to prepare intermediate Z31-a, MS m / z (ESI): 459.2 [M+H] + .

[0307] Step 2: Z25-f (130 mg, 0.249 mmol), Z31-a (185 mg, 0.374 mmol), HATU (104 mg, 0.274 mmol), DMF (1 mL), THF (1 mL) and triethylamine (0.07 mL, 0.498 mmol) were added to the reaction bottle at room temperature, and stirred at room temperature for 3 hours. Water (10 mL) was added to the reaction solution, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated and purified by column chromatography (DCM / MeOH=300 / 1-20 / 1) to obtain intermediate Z31 (130 mg, light yellow oil, purity 95.2%), yield 54.4%, MS m / z (ESI): 962.4 [M+H] + .

[0308] Preparation of intermediate Z32

[0309]

[0310] Step 1: Z25-f (150 mg, 0.288 mmol) and Z3 (150 mg, 0.345 mmol) were added to a mixture of DMF and THF (v / v=1:1, 4 mL) at room temperature, the reaction solution was cooled to 0°C, HATU (131.1 mg, 0.345 mmol) was added, and triethylamine (0.04 mL, 0.28 mmol) was slowly added dropwise. After the addition was completed, the mixture was returned to room temperature for 5 h. The reaction solution was concentrated and purified by column chromatography (MeOH / DCM=1 / 30) to obtain intermediate Z32 (160 mg, white solid, purity 100%), with a yield of 61.5%, MS m / z (ESI): 902.4 [MH] - .

[0311] Preparation of intermediate Z33

[0312]

[0313] Step 1: At room temperature, add intermediate Z4 (146 mg, 0.345 mmol), Z25-f (150 mg, 0.287 mmol), HATU (131 mg, 0.345 mmol), DMF (1 mL) and THF (1 mL) to the reaction bottle in sequence, add triethylamine (0.08 mL, 0.574 mmol) dropwise to the reaction system, and stir at room temperature for 2 h. The reaction solution was concentrated and purified by column chromatography (PE / EA=200 / 1-20 / 1) to obtain intermediate Z33 (230 mg, light yellow solid, purity 97%), with a yield of 89.8%, MS m / z (ESI): 890.4 [M+H] + .

[0314] Example 1: Preparation of Compound 1

[0315]

[0316] Step 1: Disperse 1a (4.0 g, 6.5 mmol) and tert-butyldimethylsilyl acetaldehyde (1.35 g, 7.8 mmol) in DMF (100 mL) at room temperature, add NaBH4 (288.6 mg, 7.8 mmol) in batches under stirring at room temperature, and react the reaction solution at room temperature for 5 h after the addition is completed. The reaction solution is extracted with ethyl acetate (100 mL × 3), and the combined organic phase is washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH = 20 / 1) to obtain 1b (1.8 g, white solid, purity 96%), yield: 62%, MS m / z (ESI): 661.1 [M+H] + .

[0317] Step 2: Disperse 1b (1.8 g, 2.7 mmol) and triethylamine (327 mg, 3.24 mmol) in THF (100 mL) at room temperature, add methanesulfonyl chloride (371 mg, 3.24 mmol) in batches under stirring at room temperature, and react at room temperature for 1 h after the addition. The reaction solution is extracted with ethyl acetate (100 mL × 3), and the combined organic phase is washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, and concentrated to obtain 1c (1.5 g, yellow solid, purity 92%), with a yield of 75%, MS m / z (ESI): 740.1 [M+H] + .

[0318] Step 3: Disperse 1c (200 mg, 0.27 mmol) and 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (74 mg, 0.27 mmol) in DMF (50 mL) at room temperature, add K2CO3 (75 mg, 0.54 mmol) in batches under stirring at room temperature, and after the addition is completed, the reaction solution is heated to 80°C for 1 h. The reaction solution is extracted with ethyl acetate (100 mL×3), the organic phases are combined, washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH=20 / 1) to obtain 1d (80 mg, white solid, purity 97%), with a yield of 32%, MS m / z (ESI): 917.1 [M+H] + .

[0319] Step 4: Disperse 1d (80 mg, 0.09 mmol) in a methanol solution of hydrogen chloride (10 mL, 4 mol / L) at room temperature, add the mixture, and react at room temperature for 1 hour. After the reaction solution is concentrated, it is purified by preparative HPLC (preparative column: 21.2×250 mm C18 column; system: 0.04% HCOOH / H2O; wavelength: 254 / 214 nm; gradient: 19%-48% acetonitrile change) to obtain compound 1 (2.0 mg, purity 99%), with a yield of 2.8%, MS m / z (ESI): 813.1 [M+H] + ; 1H NMR(400MHz,DMSO-d6)δ7.81(s,1H),7.58(s,2H),7.30(s,1H),7.16(s,5H),6.78(s,2H),5 .04(s,1H),4.82(s,1H),4.36(s,1H),4.25(s,2H),3.87(s,2H),3.64(s,1H),3.06(d,J=15 .1Hz,2H),2.92–2.81(m,2H),2.74(s,1H),2.65(s,2H),2.30(s,1H),1.97(d,J=7.4Hz,2H) ,1.72(s,1H),1.60(s,1H),1.48(d,J=11.8Hz,2H),1.07(s,1H),1.03(s,1H),0.83(s,1H).

[0320] Example 2: Preparation of Compound 2

[0321]

[0322] Step 1: Disperse 1c (200 mg, 0.27 mmol) and 3-(5-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione (70 mg, 0.27 mmol) in DMF (5 mL) at room temperature, add K2CO3 (75 mg, 0.54 mmol) in batches under stirring at room temperature, and after the addition is complete, the reaction solution is reacted at 80°C for 1 h. The reaction solution is extracted with ethyl acetate (100 mL×3), the organic phases are combined, washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH=20 / 1) to obtain 2a (66 mg, white solid, purity 92%), with a yield of 27%, MS m / z (ESI): 902.1 [M+H] + .

[0323] Step 2: Disperse 2a (66 mg, 0.07 mmol) in a methanol solution of hydrogen chloride (10 mL, 4 mol / L) at room temperature. After the addition was completed, the reaction solution was reacted at room temperature for 1 h. The reaction solution was concentrated and purified by preparative HPLC (preparative column: 21.2×250 mm C18 column; system: 0.04% HCOOH / H2O; wavelength: 254 / 214 nm; gradient: 19%-35% acetonitrile change) to obtain compound 2 (4.0 mg, purity 100%), yield 5%, MS m / z (ESI): 798.1 [M+H] + ; 1H NMR (400MHz, DMSO-d6) δ10.21(t,J=5.9Hz,1H)7.82(t,J=5.9Hz,1H),7.57(s,1H),7.29(d,J=5.6Hz,1H),7.19–7.14(m,4H),6.91 (d,J=7.4Hz,1H),6.80(s,1H),6.77(s,1H),5.35(s,2H),5.13(dd,J=13.5,5.2Hz,1H),4.82(s,2H),4.23(s,2H),4.19–4.04(m,2H ),3.85(s,1H),3.83(s,2H),3.15(s,2H),3.06(d,J=15.8Hz,2H),2.98–2.88(m,2H),2.73(d,J=14.7Hz,1H),2.63(d,J=16.2Hz,1 H), 2.32–2.20 (m, 2H), 1.98 (d, J = 7.7Hz, 2H), 1.72 (t, J = 11.0Hz, 1H), 1.61 (t, J = 10.4Hz, 1H), 1.48 (d, J = 12.4Hz, 1H), 1.12 (s, 1H).

[0324] Example 3: Preparation of Compound 3

[0325]

[0326] Step 1: Disperse tert-butyl 2-amino-7,7-difluoro-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (5.0 g, 17.16 mmol) and succinic acid monomethyl chloride (3.88 g, 25.75 mmol) in THF (100 mL) at room temperature. Add sodium hydride (1.03 g, 60 wt%, 25.75 mmol) in batches under stirring at room temperature. After the addition is completed, the reaction system is evacuated and replaced with argon three times, and the reaction solution is heated to 60°C for 5 h. The reaction solution was slowly added to a saturated aqueous ammonium chloride solution (100 mL), extracted with ethyl acetate (100 mL × 3), and the organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE / EA = 5 / 1) to obtain 3a (4.33 g, white solid, purity 85%), with a yield of 62%, MS m / z (ESI): 406.1 [M+H] + .

[0327] Step 2: Add 3a (4.7 g, 11.59 mmol) to 1,4-dioxane (47 mL) at room temperature, add hydrogen chloride / dioxane solution (29 mL, 4 mol / L) dropwise under stirring at room temperature, and react at room temperature for 4 h. Filter the reaction solution, wash the filter cake with dioxane, and dry the filter cake to obtain 3b (2.62 g, white solid), with a yield of 74%. 1 H NMR (400MHz, DMSO-d6) δ12.62(s,1H),10.57(br,2H),4.44(s,2H),4.05-3.89(m,2H),3.59(s,3H),2.74(t,J=6.4Hz,2H),2.66(t,J=6.4Hz,2H).

[0328] Step 3: K2CO3 (552.59 mg, 4.00 mmol) was added to a DMF (5 mL) solution containing 2-amino-3-carboxylic acid-6-chloropyrazine (300 mg, 1.73 mmol) and (R)-N-((S)-1,3-dihydrospiro[indene-2,4'-piperidin]-1-yl)-2-methylpropane-2-sulfenamide (530 mg, 1.73 mmol), and the reaction solution was stirred at 80°C for 2 h. The reaction solution was concentrated and purified by silica gel column chromatography (DCM: MeOH = 5:1) to give 3f (398 mg, white solid); yield 51.2%, MS m / z (ESI): 444.2 [M+H] + .

[0329] Step 4: 3b (1.07 g, 3.13 mmol), 3f (1.39 g, 3.13 mmol), and HATU (1.43 g, 3.76 mmol) were added to DMF (20 mL) at room temperature, and DIEA (1.21 g, 9.39 mmol) was added dropwise under stirring at room temperature. After the addition was completed, the mixture was reacted at room temperature for 2 h. The reaction solution was slowly added to water (100 mL), stirred at room temperature for 2 h, filtered, and the filter cake was washed with water. The filter cake was dried to obtain 3c (2.16 g, light yellow solid, 91% purity, yield: 94%). MS m / z (ESI): 731.3 [M+H] + .

[0330] Step 5: Add 3c (2.2 g, 3.01 mmol) to THF at room temperature, add water (1 mL) and lithium hydroxide (216 mg, 9.03 mmol) under stirring at room temperature, and after the addition is complete, heat to 80°C and react for 2 hours. The reaction solution is concentrated and purified by column chromatography (DCM / MeOH=10 / 1) to obtain 3d (1.53 g, yellow solid), yield: 70%, 1H NMR (400MHz, DMSO-d6) δ12.49(br,2H),7.62(s,1H),7.28–7.20(m,4H),6.79(s,2H),5.64(d,J=10.4Hz,1H),5.01(s,2H),4.56–4.31(m,5 H),3.16–3.05(m,3H),2.74–2.64(m,3H),9.02-8.88(m,1H),2.55–2.49(m,2H),2.04–2.00(m,1H),1.69–1.57(m,2H),1.30–1.20(m,10H).

[0331] Step 6: Compound 3d (100 mg, 0.139 mmol), Z17-b (56 mg, 0.139 mmol), and HATU (64 mg, 0.167 mmol) were added to a mixed solution of DMF (1 mL) and THF (0.5 mL) at room temperature. Triethylamine (28 mg, 0.279 mmol) was added under ice-water bath conditions. After the addition was completed, the temperature was raised to 50°C and reacted for 6 hours. The reaction solution was concentrated and purified by column chromatography (DCM / MeOH=20 / 1) to obtain 3e (81 mg, light yellow solid), with a yield of 56%. 1 H NMR(400MHz,DMSO-d6)δ12.35(s,1H),11.10(s,1H),8.01–7.98(m,1H),7.85–7.81(m,1H),7.62(s,1H),7.46–7.20(m,5H),6.79(s,2H),5 .64(d,J=10.4Hz,1H),5.16–4.91(m,3H),4.66–4.11(m,7H),3.24–2.54(m,13H),2.46–2.42(m,2H),2.05–1.53(m,6H),1.39–1.11(m,9H).

[0332] Step 7: Under ice-water bath conditions, 3e (80 mg, 0.077 mmol) was added to a mixed solution of THF (1 mL) and MeOH (1 mL). Under ice-water bath conditions, hydrogen chloride / 1,4-dioxane solution (0.2 mL, 4 mol / L) was added dropwise. After the addition was completed, the reaction was restored to room temperature for 0.5 h. The reaction solution was concentrated and purified by preparative HPLC (preparative column: 21.2×250 mm C18 column; system: 0.04% HCOOH / H2O; wavelength: 254 / 214 nm; gradient: 11%-52% acetonitrile change) to obtain compound 3 (30 mg, white solid, purity 93%), yield 41%, MS m / z (ESI): 926.4 [M+H] + ;1 H NMR (400MHz, DMSO-d6) δ11.10(br,1H),8.28(s,1H),8.02–7.99(m,1H),7.83(d,J=8.4Hz,1H),7.61(s,1 H),7.41(d,J=2.0Hz,1H),7.35–7.31(m,2H),7.21–7.16(m,3H),6.80(s,2H),5.13–5.09(m,1H),5.00(s, 2H),4.95(s,2H),4.29–4.26(m,2H),4.20–4.17(m,2H),3.24–3.06(m,8H),2.93–2.83(m,1H),2.68–2.5 4(m,5H),2.46–2.42(m,2H),2.06–2.01(m,1H),1.91–1.85(m,2H),1.78–1.49(m,3H),1.15–1.12(m,1H).

[0333] Example 4: Preparation of Compound 4

[0334]

[0335] Step 1: Dissolve 1a (2.41 g, 3.91 mmol) and tert-butyl 3-formylazetidine-1-carboxylate (940.89 mg, 5.08 mmol) in DMF (8 mL), place in a 60°C oil bath and heat with stirring for 1 h. Add NaBH4 (147.83 mg, 3.91 mmol) at room temperature and continue stirring for 10 min. Concentrate the reaction solution and purify it with a Combi Flash silica gel column (basified 20 g, 0-5% MeOH / DCM) to obtain 4a (811 mg, yellow solid, purity 35%), yield 9.24%, MS m / z (ESI): 786.1 [M+H] + .

[0336] Step 2: 4a (811 mg, 361.15 μmol) was dissolved in DCM (2.5 mL), and TFA (411.78 mg, 3.61 mmol) was added, and the reaction was stirred at 20°C for 1 h. A large amount of dichloromethane was added to the reaction solution, and ethyl acetate (10 mL) was added to dissolve after concentration, and saturated sodium bicarbonate solution (50 mL) was added to neutralize, and the aqueous phase was extracted with ethyl acetate (20 mL×2), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain 4b (423 mg, yellow solid, purity 35%), with a yield of 59.77%, MS m / z (ESI): 686.3 [M+H] + .

[0337] Step 3: 4b (224 mg, 114.31 μmol) and 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (37.89 mg, 137.17 μmol) were dissolved in DMSO (3 mL), and DIEA (44.32 mg, 342.93 μmol) was added, and the mixture was stirred at 100 °C for 45 min under microwave heating. Water (100 mL) was added to dilute the mixture, and the aqueous phase was extracted with ethyl acetate (20 mL × 2). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by Combi Flash silica gel column (4 g, 0-4% MeOH / DCM) to obtain 4c (121 mg, yellow solid, yield 20.23%, purity 18%), MS m / z (ESI): 942.0 [M+H] + .

[0338] Step 4: 4c (121 mg, 23.12 μmol) was dissolved in 1,4-dioxane (4.5 mL), and hydrochloric acid (8.43 mg, 231.19 μmol, 0.1 mL) was added dropwise, and the mixture was stirred at 20°C for 5 min. The reaction solution was concentrated, and dichloromethane (10 mL) and an appropriate amount of triethylamine were added to the mixture until it was completely dissolved. After further concentration, the mixture was purified by preparative liquid chromatography (column: Waters-SunFire C18, mobile phase: 0.05% NH4HCO3 water / acetonitrile, 54% acetonitrile-62% acetonitrile, 30 mL / min) to obtain compound 4 (1.0 mg), with a yield of 5.16%, MS m / z (ESI): 838.0 [M+H] + .

[0339] Example 5: Preparation of Compound 5

[0340]

[0341] Step 1: Dissolve Z25-d (182 mg, 349.67 μmol) and 3-bromoprop-1-yne (62.40 mg, 524.51 μmol) in DMF (0.6 mL), add K2CO3 (144.98 mg, 1.05 mmol) at 20°C, and continue stirring for 3 h. The reaction solution was filtered through a diatomaceous earth layer, and the filter cake was washed with DCM (20 mL). The filtrate was concentrated and purified by a Combi Flash silica gel column (4 g, 0-40% EA / PE, 0-5% MeOH / DCM) to obtain 5a (71 mg, yellow solid, purity 88%), with a yield of 38.87%, MS m / z (ESI): 339.3 [M+120] + .

[0342] Step 2: Dissolve 5a (71 mg, 135.93 μmol) in DCM (1 mL) at room temperature, add TFA (154.99 mg, 1.36 mmol) dropwise, and continue stirring for 30 min. Add DCM (20 mL) to the reaction solution, concentrate, add DCM (30 mL) again, and concentrate to obtain 5b (55 mg, yellow oily liquid, purity 40%), yield 45.02%, MS m / z (ESI): 360.2 [M+H] + .

[0343] Step 3: Dissolve 5b (55 mg, 61.19 μmol) and (3-amino-5-chloropyrazin-2-yl)(2-amino-7,7-difluoro-6,7-dihydrothiazolo[5,4-c]pyridin-5(4H)-yl)methanone (25.46 mg, 73.43 μmol) in DMF (2 mL), add K2CO3 (25.37 mg, 183.57 μmol), and heat in an oil bath at 70°C with stirring for 2 h. The reaction solution was filtered through a celite layer, and the filter cake was washed with DCM (30 mL) and MeOH (3 mL). The filtrate was concentrated and purified by Combi Flash silica gel column (4 g, 0-40% EA / PE, 0-5% MeOH / DCM) to obtain 5c (55 mg, yellow solid, purity 44%), with a yield of 59.04%, MS m / z (ESI): 671.3 [M+H] + .

[0344] Step 4: Dissolve intermediate Z12 (16.75 mg, 43.36 μmol) and 5c (55 mg, 36.13 μmol) in THF (1.5 mL) and water (1.5 mL), then add sodium ascorbate (715.75 μg, 3.61 μmol) and CuSO4 (1.15 mg, 7.23 μmol), and stir at 20°C for 14 h. Filter the reaction solution through a diatomaceous earth layer, add DCM (30 mL) and MeOH (5 mL) to wash the filter cake, and purify the filtrate through a Combi Flash silica gel column (4 g, 0-20% EA / PE, 0-5% MeOH / DCM) to obtain 5d (57 mg, brown solid, purity 63%), yield 94.11%, MS m / z (ESI): 518.6 [(M-19+H) / 2] + .

[0345] Step 5: 5d (57 mg, 34.00 μmol) was dissolved in THF (1 mL) and MeOH (0.5 mL), and hydrochloric acid (12.40 mg, 340.00 μmol, 0.1 mL) was added, and the mixture was stirred at 20°C for 5 min. DCM (20 mL) was added to the reaction solution for dilution, and after concentration, it was purified by preparative liquid chromatography (column: Waters-SunFire C18, mobile phase: 0.05% NH4HCO3 water / acetonitrile, 5% acetonitrile-90% acetonitrile, 30 mL / min) to obtain compound 5 (4.15 mg, yellow solid, purity 98.74%), with a yield of 12.66%, MS m / z (ESI): 936.0 [M-17+H] + ; 1 H NMR (400MHz, DMSO-d6) δ8.35(s,1H),7.87(s,1H),7.57(s,1H),7.49(d,J=8.3Hz,1H),7.20(s,2H),7.01(d,J=8.0Hz,1H), 6.92(d,J=2.1Hz,1H),6.82–6.77(m,2H),6.56(s,2H),6.45(d,J=10.3Hz,2H),5.89(s,1H),5.07(dd,J=13.0,5.3Hz,1H),4 .80(s,2H),4.43(d,J=5.3Hz,2H),4.23(d,J=5.6Hz,4H),3.87–3.70(m,5H),3.42–3.37(m,4H),3.14(s,2H),2.90(d,J=16 .0Hz,2H),2.76–2.56(m,1H),2.60–2.52(m,1H),1.97(s,2H),1.61(d,J=11.3Hz,2H),1.40(d,J=12.8Hz,2H),1.22(s,2H).

[0346] Example 6-Example 17

[0347] Compound 6-Compound 17 were prepared by referring to the above preparation method, substituting the intermediates in Table 2 to obtain the corresponding compounds shown in Table 2 and Table 3.

[0348] Table 2

[0349]

[0350]

[0351] The structural characterization data of the obtained compound are shown in Table 3:

[0352] Table 3

[0353]

[0354]

[0355] Example 18: Preparation of Compound 18

[0356]

[0357] Step 1: Dissolve Z13 (230 mg, 443.59 μmol) and 3-amino-5-chloro-pyrazine-2-carboxylic acid (76.99 mg, 443.59 μmol) in THF (20 mL), add DIEA (143.33 mg, 1.11 mmol, 193.17 μL) and T3P (423.19 mg, 1.33 mmol), and react at room temperature for 1 h. The reaction solution was concentrated and purified by silica gel column chromatography (DCM: MeOH = 100: 5) to obtain 18a (210 mg, brown solid, purity 93.4%), yield: 70.23%; MS m / z (ESI): 673.9 [M+H] + .

[0358] Step 2: The reactant 18a (200 mg, 296.72 μmol), (R)-N-((S)-1,3-dihydrospiro[indene-2,4'-piperidin]-1-yl)-2-methylpropane-2-sulfenamide (118.21 mg, 385.74 μmol) and were dissolved in DMF (8 mL), K2CO3 (73.82 mg, 534.10 μmol) was added, and the reaction was heated to 80°C for 7 hours. The reaction solution was concentrated and purified by silica gel column chromatography (DCM: MeOH = 100:5) to obtain 18b (185 mg, brown solid, purity 82.1%), yield: 66.04%; MS m / z (ESI): 472.7 [M / 2+H] + .

[0359] Step 3: 18b (200 mg, 211.86 μmol) was dissolved in MeOH (6 mL) and THF (1.5 mL), and hydrogen chloride / 1,4-dioxane (1 mL, 4 mol / L) was added, and the reaction was allowed to react at room temperature for 15 minutes. The reaction solution was concentrated and separated by an acidic preparative column (preparative column: 21.2×250 mm C18 column; system: 0.04% HCOOH / H2O; wavelength: 254 / 214 nm; gradient: 24%-44% acetonitrile change) to obtain compound 18 (80 mg, purity 99.2%), yield: 44.6%; MS m / z (ESI): 840.0 [M+H] + . 1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),δ8.21(s,1H),7.68–7.49(m,2H),7.41–7.24(m,1 H),7.18(dd,J=7.4,3.7Hz,4H),7.03(d,J=7.0Hz,1H),6.78(d,J=24.4Hz,3H),5.03(dd, J=12.7,5.4Hz,1H),4.54(s,3H),4.27(d,J=13.2Hz,2H),3.64(q,J=6.7Hz,2H),3.16–2 .99(m,2H),2.89–2.56(m,5H),2.12–1.89(m,1H),1.85–1.45(m,3H),1.28–0.91(m,2H).

[0360] Example 19-Example 29

[0361] Compounds 19 to 29 can be prepared by referring to the preparation method of Example 18, replacing the intermediate with a suitable intermediate in Table 4 to obtain the corresponding compounds, as shown in Tables 4 and 5:

[0362] Table 4

[0363]

[0364]

[0365] The structural characterization data of the obtained compound are shown in Table 5:

[0366] Table 5

[0367]

[0368]

[0369] Example 30: Preparation of Compound 30

[0370]

[0371] Step 1: Disperse the intermediate Z25 (78 mg, 0.09 mmol) in DCM (5 mL) at room temperature, add TFA (1 mL) in batches under stirring at room temperature, and react the reaction solution at room temperature for 2 h. The reaction solution was extracted with ethyl acetate (100 mL × 3), and the organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH = 20 / 1) to obtain 30a (48 mg, white solid, purity 79%), yield: 69%, MS m / z (ESI): 734.0 [M+H] + .

[0372] Step 2: 30a (48 mg, 0.065 mmol) and (3-amino-5-chloropyrazin-2-yl)(2-amino-7,7-difluoro-6,7-dihydrothiazolo[5,4-c]pyridin-5(4H)-yl)methanone (22 mg, 0.065 mmol) were dispersed in DMF (5 mL) at room temperature. K2CO3 (9 mg, 0.065 mmol) was added in batches under stirring at room temperature. The reaction solution was reacted at 80°C for 2 h. The reaction solution was extracted with ethyl acetate (100 mL×3). The organic phases were combined, washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH=20 / 1) to obtain 30b (30 mg, white solid, purity 76%), yield: 44%, MS m / z (ESI): 522.0[(M+H) / 2] + .

[0373] Step 3: Disperse 30b (30 mg, 0.03 mmol) in hydrogen chloride / 1,4-dioxane (5 mL, 4 mol / L) at room temperature, and react the reaction solution at room temperature for 2 h. After the reaction solution was concentrated, it was purified by preparative liquid phase (preparative column: 21.2×250 mm C18 column; system: 0.04% HCOOH / H2O; wavelength: 254 / 214 nm; gradient: 13%-37% acetonitrile change) to obtain compound 30 (3.5 mg, purity 100%), yield: 19%, MS m / z (ESI): 940.0 [M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ9.86(s,1H),7.94(s,1H),7.58(s,1H),7.54–7.47(m,2H),7.28(s,1H),7.20(s,2H),7.17(s, 1H),7.09(s,1H),6.93(s,1H),6.83–6.77(m,2H),5.06–4.95(m,1H),4.81(s,2H),4.51–4.37(m,1H),4.23(s,2H),3.8 3(s,1H),3.44(s,2H),3.14(s,4H),2.97(s,2H),2.83(d,J=11.9Hz,2H),2.56(s,2H),2.52(s,2H),2.39(d,J=6.8Hz, 2H),2.22(s,1H),1.97(d,J=7.5Hz,2H),1.68(s,2H),1.60–1.55(m,1H),1.43(s,1H),1.15–1.09(m,1H),0.83(s,1H).

[0374] Example 31: Preparation of Compound 31

[0375]

[0376] Step 1: Disperse intermediate Z26 (61 mg, 0.07 mmol) in DCM (5 mL) at room temperature, add TFA (1 mL) in batches under stirring at room temperature, and after the addition is complete, the reaction solution is reacted at room temperature for 2 h. The reaction solution is concentrated and purified by column chromatography (DCM / MeOH=20 / 1) to obtain 31a (51 mg, white solid, purity 71%), yield: 91%, MS m / z (ESI): 817.0 [M+H] + .

[0377] Step 2: 31a (51 mg, 0.063 mmol) and (3-amino-5-chloropyrazin-2-yl)(2-amino-7,7-difluoro-6,7-dihydrothiazolo[5,4-c]pyridin-5(4H)-yl)methanone (21 mg, 0.063 mmol) were dispersed in DMF (5 mL) at room temperature. K2CO3 (9 mg, 0.063 mmol) was added in batches under stirring at room temperature. The reaction solution was reacted at 80°C for 2 h. The reaction solution was extracted with ethyl acetate (100 mL×3), and the organic phases were combined, washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM / MeOH=20 / 1) to obtain 31b (45 mg, white solid, purity 70%), yield: 64%, MS m / z (ESI): 564.0 [M / 2+H] + .

[0378] Step 3: Disperse 31b (45 mg, 0.04 mmol) in hydrogen chloride / 1,4-dioxane (5 mL, 4 mol / L) at room temperature, add and react at room temperature for 2 h. After the reaction solution is concentrated, it is purified by preparative liquid phase (preparative column: 21.2×250 mm C18 column; system: 0.04% HCOOH / H2O; wavelength: 254 / 214 nm; gradient: 11%-40% acetonitrile change) to obtain compound 31 (7.0 mg, white solid, purity 100%), yield: 16%, MS m / z (ESI): 512.0 [M / 2+H] + ; 1H NMR (400MHz, DMSO-d6) δ11.07(s,1H),9.88(s,1H),7.59(s,1H),7.55(d,J=8.4Hz,1H),7.50(s,1H),7.31(d,J=8.0Hz,1H),7.21(s,2H),7 .15(s,1H),6.95(s,1H),6.85(d,J=8.9Hz,1H),6.79(s,2H),5.30(s,1H),5.01(dd,J=13.0,5.5Hz,1H),4.81(s,2H),4.46(s,2H),4.23(s ,2H),3.85(s,1H),3.47(s,5H),3.45(s,4H),3.15(d,J=11.6Hz,2H),3.02(d,J=16.0Hz,1H),2.84(d,J=12.2Hz,1H),2.64(d,J=7.5Hz,4H ), 2.53(s,2H),2.31(s,1H),2.21(s,1H),1.98(d,J=8.0Hz,2H),1.64(d,J=33.2Hz,2H),1.46(d,J=11.0Hz,2H),1.14(s,1H),0.83(s,1H).

[0379] Example 32-Example 38

[0380] Compounds 32 to 38 can refer to the preparation method of Example 30 or Example 31, and the intermediates are replaced with appropriate intermediates in Table 6 to obtain the corresponding compounds, as shown in Table 6 and Table 7:

[0381] Table 6

[0382]

[0383]

[0384] The structural characterization data of the obtained compound are shown in Table 7:

[0385] Table 7

[0386]

[0387]

[0388] Test Example 1: Phosphatase activity detection

[0389] IC 50The value was detected by Homogeneous Full Length SHP-2Assay Kit (BPS Bioscience #79330). The kit uses 6,8-difluoro-4-methylumbelliferyl phosphate (DiFMUP) as the reaction substrate. SHP2 enzyme solution (diluted to 0.04ng / μL with 1× buffer) and 0.5μM SHP-2 activating peptide (activating peptide) were mixed in 1× buffer solution containing 2.5mM DTT to activate PTP. DMSO (0.1% (V / V)) or compound (concentration: 0.5nM-1μM) was added and mixed and incubated for 1h. DiFMUP (50μM, total reaction volume of 20μL) was added to start the reaction. After incubation for 30min, the fluorescence intensity of the reaction solution was detected by Tecan Infinite M1000 (excitation light: 350nm, emission light 460nm). IC was calculated by XLfit 5.0 software 50 The test results are shown in Table 8. From the results in Table 8, it can be seen that the compounds provided in the present application have good inhibitory activity against SHP-2 enzyme.

[0390] Table 8: IC of compounds for inhibition of SHP-2 enzyme activity 50 value

[0391]

[0392]

[0393] Test Example 2: Cell Proliferation Detection

[0394] MV 411 cells (20,000 cells / well) were inoculated in a 96-well plate (Corning #3916) and cultured in 80 μL / well of culture medium (10% fetal bovine serum (FBS) culture medium (IMDM), Gibco #10099-141C). After 24 hours of culture, 20 μL of the compound prepared in this application at different concentrations was added (the final concentration of DMSO was 0.1% (V / V)). On the 4th day, 60 μL of CellTiter-Glo (Promega #G7558) was added to each well at 80 rpm, 25°C, 15 min, and the fluorescence value (i.e., signal value) was detected using Victor X5 (PE). IC was calculated by XLfit 5.0 software 50 The test results are shown in Table 9. As can be seen from Table 9, the compounds provided in the present application have significant inhibitory activity on MV-4-11 cell proliferation.

[0395] Table 9: IC values ​​of compounds for MV-4-11 cell proliferation inhibition 50 value

[0396] Compound No. <![CDATA[IC 50 (μM)]]> Compound No. <![CDATA[IC 50 (μM)]]> Compound No. <![CDATA[IC 50 (μM)]]> 1 0.0632 9 0.5783 21 0.0349 2 0.0814 13 0.2222 31 0.0625 3 0.044 15 0.0483 37 0.1587 6 0.0563 18 0.0586

[0397] Test Example 3 Protein degradation test

[0398] 1. Compound Treatment of MV411 Cells

[0399] 1) Collect logarithmic phase MV411 cells and adjust the cell suspension concentration to 8×e 5 / mL, the cells were plated into 12-well plates, 1 mL of 5% CO2 (volume fraction, the rest was air) was added to each well and incubated at 37°C.

[0400] 2) Add different concentrations of compounds to each well to the final concentrations of 1.11 μM, 0.37 μM, 0.12 μM, and 0.04 μM, add 0.1% DMSO as blank wells, and incubate at 5% CO2 and 37°C for 16 h.

[0401] 2. Collecting Protein Samples

[0402] 1) Collect the treated cells into a 1.5 mL centrifuge tube, centrifuge at 300 g for 5 min, wash once with PBS, and discard the PBS.

[0403] 2) After adding 50 μL of 1× cell lysis buffer to each sample and mixing with the cells, place on ice for 10 min, centrifuge at 12000 rpm for 10 min, collect the supernatant, and store in a -80°C refrigerator.

[0404] 3) Take 32.5 μL of supernatant, add 17.5 μL of 4× loading buffer containing DTT, mix well and boil the sample in a metal bath at 100°C for 5 min.

[0405] 4) Electrophoresis: Place the 26-well precast gel in the electrophoresis tank, add buffer, load 10 μL of sample into each well, turn on the power supply, 60V, adjust the voltage to 100V after 30 minutes and continue electrophoresis. Terminate the electrophoresis when the bromophenol blue has almost completely run out of the PAGE gel;

[0406] 5) Transfer: After electrophoresis, remove the gel and place it in a transfer sandwich, then place it in a transfer apparatus at 25V for 7min.

[0407] 6) Blocking: After the transfer, immerse the NC membrane in TBST buffer containing 5% BSA and shake it on a shaker at room temperature for 1 hour.

[0408] 7) Primary antibody incubation: After blocking, wash 3 times with TBST buffer, then add primary antibody at a dilution ratio of 1:1000 and incubate overnight at 4°C. After incubation, wash the NC membrane 3 times with TBST buffer, shaking for 5 minutes each time.

[0409] 8) Secondary antibody incubation: Discard the TBST buffer, add 1:1000 secondary antibody, and shake at room temperature for 1 hour. Discard the secondary antibody and wash the membrane 3 times with TBST buffer, shaking for 5 minutes each time.

[0410] 9) Exposure: Evenly cover the ECL color developing substrate on the NC film and expose it for imaging.

[0411] 3. Experimental results

[0412] Figure 1 The figure is a Western blot (immunoblotting test), i.e., the result of the degradation of SHP2 protein by the compound of formula (I); wherein Actin represents the internal reference. Figure 1 It can be clearly seen that the exemplary compounds of the present application, such as Compound 11, Compound 12, Compound 13, Compound 14, Compound 16, Compound 17, Compound 35, and Compound 38, have a degradation effect on SHP2 protein, which indicates that the compounds of the present application have a good degradation effect on SHP2 protein.

[0413] All documents mentioned in this application are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teaching content of this application, those skilled in the art can make various changes or modifications to this application, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound represented by formula (I), or a pharmaceutically acceptable salt thereof: in, R1 and R2 are each independently hydrogen, C 1-3 Alkyl, -NH2, C 1-3 Alkoxy, cyano, hydroxy, carboxyl, halogen (preferably fluorine or chlorine), halogenated C 1-3 Alkyl, halogenated C 1-3 Alkoxy, -C(O)NH2, -C(O)C 1-3 Alkyl, -OC(O)C 1-3 Alkyl or -C(O)OC 1-3 alkyl; Preferably, R1 and R2 are each independently hydrogen, methyl, -NH2, methoxy, cyano, hydroxyl, carboxyl, fluorine, chlorine, trifluoromethoxy, difluoromethoxy, monofluoromethoxy, trifluoromethyl, difluoromethyl, monofluoromethyl, -C(O)NH2, -COCH3, -COOCH3 or -OCOCH3; Preferably, R1 is hydrogen; Preferably, R2 is -NH2; R3 and R4 are each independently hydrogen, C 1-3 Alkyl or -C(O)C 1-6 alkyl; Preferably, R3 and R4 are each independently hydrogen, methyl or -C(O)CH3; Preferably, R3 and R4 are hydrogen; (R5) m1 represents m1 R5, m1 is 0 or 1, each R5 is the same or different, and each independently represents halogen (preferably fluorine, chlorine or bromine), cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, -NH2, -NHCOC 1-3 Alkyl, -C(O)C 1-3 Alkyl, -C(O)OC 1-3 Alkyl, -OC(O)C 1-3 Alkyl, hydroxyl, carboxyl, halogenated C 1-3 Alkyl or halogenated C 1-3 Alkoxy; Preferably, (R5) m1 represents m1 R5, m1 is 0 or 1, each R5 is the same or different, and each independently represents fluorine, chlorine, cyano, methyl, methoxy, vinyl, ethynyl, -NH2, -NHCOCH3, -C(O)CH3, -C(O)OCH3, -OC(O)CH3, hydroxyl, carboxyl, trifluoromethyl, difluoromethyl, monofluoromethyl, trifluoromethoxy, difluoromethoxy or monofluoromethoxy; Preferably, m1 is 1, and R5 is -NH2; Preferably, m1 is 0; (R6) m2 represents m2 R6, m2 is 0, 1, 2, 3 or 4, each R6 is the same or different, and each independently represents halogen (preferably fluorine, chlorine or bromine), cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl or halogenated C 1-3 Alkoxy; Preferably, (R6) m2 represents m2 R6, m2 is 0, 1, 2, 3 or 4, each R6 is the same or different, and each independently represents fluorine, chlorine, cyano, methyl, methoxy, vinyl, ethynyl, trifluoromethyl, difluoromethyl, monofluoromethyl, trifluoromethoxy, difluoromethoxy or monofluoromethoxy; Preferably, R6 is halogen, preferably fluorine, chlorine or bromine; Preferably, m2 is 2, and R6 is fluorine; W is -CH2- or -C(O)-; X is -(X1) p1 -(L1) q1 -(X2) p2 -(L2) q2 -(X3) p3 -(L3) q3 -(X4) p4 -(L4) q4 -, Wherein, L1, L2, L3, and L4 are each independently C 1-10 Alkylene; X1, X2, X3, and X4 are each independently -O-, -NH-, -C(O)-, -NHC(O)-, or -(OCH2CH2) p5 -, azetidine ring group, tetrahydropyrrole ring group, piperazine ring group, piperidine ring group, triazole ring group, C 2-4 Alkenylene, C 2-4 Alkyne group, p1, p2, p3, p4, p5, q1, q2, q3, q4 are each independently 0, 1, 2, 3 or 4; Preferably, L1, L2, L3, L4 are each independently -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4- or -(CH2)7-; Preferably, X1, X2, X3, and X4 are each independently -O-, -NH-, -C(O)-, -NHC(O)-, -(OCH2CH2)1-, -(OCH2CH2)2-, -(OCH2CH2)3-, -(OCH2CH2)4-, Preferably, the compound represented by formula (I) has the structure represented by formula (I-1), 2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that structure for Preferably, the structure for 3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that structure for:

4. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that X is selected from the following structures: Wherein, n is each independently an integer of 1-10; Preferably, X is selected from the following structures: Preferably, X is selected from the following structures: in End and Connection, "-*" end and connect; Preferably, X is selected from the following structures, in End and Connection, "-*" end and connect.

5. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The compound of formula (I) has the structure shown in formula (II), Wherein, X is selected from the following structures: Wherein, n is each independently an integer of 1-10; Preferably, X is selected from the following structures: Wherein, n is each independently an integer of 1-5; Preferably, X is selected from the following structures: in End and Connection, "—*" end and connect; Preferably, the compound of formula (II) is a structure represented by formula (II-1), 6. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The compound of formula (I) has the structure shown in formula (III), Wherein, X is selected from the following structures: Wherein, n is each independently an integer of 1-10; Preferably, X is selected from the following structures: Wherein, n is each independently an integer of 1-5; Preferably, X is selected from the following structures: in End and Connection, "—*" end and connect; Preferably, the compound of formula (III) is a compound of formula (III-1), 7. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The compound of formula (I) is selected from the following compounds:

8. A pharmaceutical composition, characterized in that The invention comprises a compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

9. Use of the compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 8 in the preparation of a medicament for treating and / or preventing a disease or condition mediated by SHP2 or associated with abnormal SHP2 activity.

10. The use according to claim 9, characterized in that: The diseases or conditions associated with abnormal SHP2 activity include: solid tumors and hematological tumors; preferably cancer; more preferably juvenile myelomonocytic leukemia, acute myeloid leukemia, B-cell acute lymphoblastic leukemia, neuroblastoma, esophageal cancer, breast cancer, lung cancer, colon cancer, gastric cancer or head and neck cancer.

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