Preparation method and application of FLT3 degradation agent based on pyrazinecarboxamide skeleton

Abstract

The invention provides a pyrazinecarboxamide compound with a structure as shown in a formula (I), or a pharmaceutically acceptable salt, a stereoisomer, an N-oxide, a prodrug molecule, a solvate and a deuterated compound of the pyrazinecarboxamide compound, a composition containing the compound as shown in the formula (I), and a pyridopyrimidone compound with the structure as shown in the formula (I). The invention also relates to application of the compound shown in the formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, or an N-oxide, a solvate and a deuterated compound thereof, and a composition containing the compound shown in the formula (I) in drugs for treating FLT3 (FMS-like tyrosine kinase 3, Fmslide tyrosine kinase) related diseases participating in cancer cell growth. The compound provided by the invention can efficiently degrade FLT3 protein in cells, so that the proliferation of tumor cells with FLT3 mutation can be remarkably inhibited, and the compound can be used for preparing drugs for treating FLT3 protein related cancers and other related diseases. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, and in particular to a class of compounds for targeting ubiquitination and degradation of FLT3 protein, and a pharmaceutical composition and application thereof. Background Art

[0002] Acute myeloid leukemia (AML) is a malignant proliferative disease caused by clonal expansion and differentiation loss of bone marrow hematopoietic stem cells, with a high relapse rate and poor prognosis. Currently, the treatment of AML mainly relies on chemotherapy combined with allogeneic hematopoietic stem cell transplantation, but elderly and frail patients have difficulty tolerating conventional chemotherapy. Targeted therapy has gradually become a hot topic in research and treatment due to its powerful advantages. Therefore, it is necessary to develop more effective targeted drugs to meet the unmet needs in the clinical treatment of AML.

[0003] The sequencing results of AML patients by the Cancer Genome Atlas Research Network showed that genes involved in the onset of the disease include FLT3 (25%-35%), NPM1 (27%), DNMT3A (26%), IDH1 / IDH2 (20%), NRAS / KRAS (12%), TP53 (8%), etc. Among them, human FMS-like tyrosine kinase 3 (FLT3) is a type III receptor tyrosine kinase located on chromosome 13q12, consisting of five immunoglobulin-like domains in the extracellular region, a juxtamembrane domain (JMD) and a tyrosine kinase (TK) domain. FLT3 is mainly expressed on the cell membrane of early hematopoietic progenitor cells and stem cells. Normally, FLT3 is silent and is not activated in normal differentiated cells. When FLT3 ligand (FL) binds to the FLT3 extracellular domain, the receptor tyrosine kinase dimerizes and then activates the downstream signaling pathway through autophosphorylation. 35% of AML patients can develop FLT3 mutations, inducing the activation of constitutive kinases independent of FL. Mutations include two main types, namely internal tandem repeat mutations in the juxtamembrane region (FLT3-ITD) and point mutations or deletions and insertion mutations in the tyrosine kinase domain (FLT3-TKD). Mutations can cause activation of multiple downstream signal transduction pathways, leading to DNA damage and repair defects, and increasing the abnormal proliferation and anti-apoptosis ability of leukemia cells. AML with FLT3 mutations is considered to be a refractory leukemia and is associated with a poor prognosis. Although the approval of FLT3 inhibitors such as midostaurin, gilteritinib and quizartinib has shown good clinical efficacy for mutated AML, the rapid emergence of drug resistance has led to a short duration of treatment and an inability to maintain the ideal therapeutic effect.

[0004] Proteolysis Targeting Chimeras (PROTAC) is a bifunctional structural molecule that degrades disease-related target proteins by inducing the ubiquitin-proteasome system (UPS). PROTAC molecules generally consist of three parts: target protein binding ligand, Linker, and E3 ligase ligand. Its working principle is that after the PROTAC molecule enters the cell, the POI ligand specifically recognizes the target protein, and the E3 ligand binds to the E3 ubiquitin ligase to form a ternary complex of POI-PROTAC-E3Ligase, which shortens the distance between the protein to be degraded and the E3 ubiquitin ligase, so that the ubiquitin on the E2 ubiquitin conjugating enzyme is transferred to the target protein and ubiquitinated, thereby triggering the proteasome to degrade the target protein and achieve the purpose of treating the disease. Compared with traditional kinase inhibitors, PROTAC technology has many advantages: 1) Inducing degradation of target proteins in a catalytic manner, that is, when the target protein is degraded, the degrader will dissociate from the complex and enter the next round of catalytic cycle; 2) Easier to achieve selectivity, that is, the sensitivity of E3 ubiquitin ligases to substrate protein degradation is different, giving degraders better selectivity; 3) Broader pharmacological functions, that is, by degrading intact proteins, thereby inhibiting kinase function and non-kinase function at the same time. Due to its many advantages in drug development, PROTAC technology has been widely used in the study of disease-related proteins, such as kinases, nuclear receptors, G protein-coupled receptors and transcription factors.

[0005] Therefore, there is an urgent need in the art to develop a compound that is suitable for treating acute myeloid leukemia and can ubiquitinate and degrade FLT3 protein. Summary of the invention

[0006] The purpose of the present invention is to provide a compound capable of targeting ubiquitination and degradation of FLT3 protein and a pharmaceutical composition thereof.

[0007] In the first aspect of the present invention, a pyrazinecarboxamide compound, a pharmaceutically acceptable salt thereof, a stereoisomer thereof or a prodrug molecule thereof is provided, wherein the compound has a structure shown in formula (I):

[0008]

[0009] in:

[0010] Y is selected from the group consisting of -CH2-, -C(=O)-;

[0011] R1 and R2 are each independently selected from the following group: -H, -F;

[0012] L has the structure shown below: -Z-(L1) m -;

[0013] wherein m is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30;

[0014] The Z is selected from the following group: a chemical bond, a 3-12 membered ring having 0-4 heteroatoms, -NH-, -O-, -C(O)-, -CONH-, -NHCO-, -S-, -CH2-, -C≡C-, -CH=CH-, -P=O-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-;

[0015] Each L1 is independently selected from the following group: a 3-12 membered ring having 0-4 heteroatoms, -NH-, -O-, -C(O)-, -CONH-, -NHCO-, -S-, -CH2-, -C≡C-, -CH=CH-, -P=O-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-;

[0016] The L segment may be optionally replaced by one or more R L Substituted; wherein said R L Selected from the following group: halogen, -OH, -CN, -CF3, -NH2, -COOH, oxygen atom (=O), substituted or unsubstituted C1-C6 alkyl or haloalkyl, substituted or unsubstituted C1-C6 alkoxy; or, 2 R L The radicals and the atoms to which they are attached together form a 3-8 membered ring containing 0-4 heteroatoms;

[0017] Ring A is a substituted or unsubstituted 5-7 membered heterocyclic ring;

[0018] L2 is a 4-7 membered heterocyclic ring or is absent;

[0019] The substitution refers to that one or more H atoms on the group are replaced by a substituent selected from the group consisting of halogen and deuterium atom.

[0020] In some embodiments, the compound of formula I has a structure selected from the following group:

[0021]

[0022] Wherein, the definitions of Y, L, R1 and R2 are as described above.

[0023] In some embodiments, L has the structure shown in the following formula:

[0024]

[0025] in:

[0026] Z is selected from the following group: a 3-8 membered ring having 0-4 heteroatoms, -NH-, -O-, -C(O)-, -CONH-, -NHCO-, -S-, -CH2-, -C≡C-, -CH=CH-, -P=O-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-;

[0027] n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0028] And the dashed line indicates the point of attachment of the linker moiety.

[0029] In some embodiments, L has the structure shown in the following formula:

[0030]

[0031] in:

[0032] W1 and W2 are absent or are each independently a 3-12 membered ring having 0-4 heteroatoms, which is optionally replaced by R L replace;

[0033] R L The definition of is as mentioned above;

[0034] B is selected from the following group: a 3-12 membered ring that is absent or has 0-4 heteroatoms, -NH-, -O-, -CONH-, -NHCO-, -S-, -C≡C-, -CH=CH-, -P=O-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-, C1-C6 alkyl or haloalkyl (straight chain, branched, optionally substituted) and optionally one or more C atoms are replaced by O, N; or C1-C6 alkoxy (straight chain, branched, optionally substituted);

[0035] Z is selected from the following group: a 3-12 membered ring containing 0-4 heteroatoms, -NH-, -O-, -CONH-, -NHCO-, -S-, -CH2-, -C≡C-, -CH=CH-, -P=O-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-;

[0036] each n is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; and

[0037] Dashed lines indicate points of attachment of linker moieties.

[0038] In some embodiments, or its stereoisomers or prodrug molecules, it is characterized in that the Z is selected from the following groups: -NHCO-, -CH2-, -C(O)-, a 3-12 membered ring containing 0-4 heteroatoms, -NH-, -O-.

[0039] In another preferred embodiment, the 3-12 membered ring having 0-4 heteroatoms is selected from the following group:

[0040] In some embodiments, L has a structure selected from the following group:

[0041]

[0042]

[0043]

[0044]

[0045]

[0046]

[0047] wherein n and m are each independently 0, 1, 2, 3, 4, 5 or 6.

[0048] In some embodiments, the compound is selected from the following group:

[0049]

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056]

[0057]

[0058] In the second aspect of the present invention, a pharmaceutical composition for treating tumors is provided, characterized in that it comprises the compound or its pharmaceutically acceptable salt, or stereoisomer, its deuterated product or its prodrug molecule as described in the first aspect of the present invention, and an optional pharmaceutically acceptable carrier.

[0059] In the third aspect of the present invention, there is provided a use of the compound as described in the first aspect of the present invention, its pharmaceutically acceptable salt, its stereoisomer, its deuterated product or its prodrug molecule, characterized in that it is used to treat or prevent a disease or condition related to the activity or expression level of FLT3; wherein the disease or condition is selected from the following group: cardiovascular disease, diabetes, hypertension, muscular dystrophy, Parkinson's disease, Alzheimer's disease, and cancer.

[0060] In some embodiments, the cancer includes but is not limited to: hematological tumors, gastrointestinal stromal tumors, histiocytic lymphoma, non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, nasopharyngeal carcinoma.

[0061] In another preferred embodiment, the invention is used to treat an indication selected from the group consisting of non-small cell lung cancer, malignant melanoma, prostate cancer, kidney cancer, bladder cancer, ovarian cancer, colon cancer, rectal cancer, breast cancer, cervical cancer, lung cancer, laryngeal cancer, nasopharyngeal cancer, pancreatic cancer, multiple myeloma, B lymphoma, leukemia, or to prevent tumor recurrence after surgery.

[0062] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0063] Figure 1 .Preliminary screening of PROTACs degradation in MV4-11 cells. 7-13 are the original experimental numbers of 2-7aa.

[0064] Figure 2 .Study on the concentration-dependent degradation of FLT3 by the preferred compounds. The activity data were all carried out in three independent repeated experiments, and the half degradation concentration (DC 50 ) = mean ± standard deviation (SD).

[0065] Figure 3 . The preferred compounds were tested for their inhibitory activity on MV4-11 cells. The activity data were obtained by three independent repeated experiments, and the half inhibitory concentration (IC 50 ) = mean ± standard deviation (SD).

[0066] Figure 4.Selective degradation study of compound 2-7aa.

[0067] Figure 5 Apoptosis analysis of MV4-11 cells after treatment with compound 2-7aa and gilteritinib.

[0068] Figure 6 .Cell cycle analysis of MV4-11 cells after treatment with compound 2-7aa and gilteritinib.

[0069] Figure 7 . Mouse xenograft tumor experiment. 7-13 is the original experimental number of 2-7aa. *P<0.05, **P<0.01, ***P<0.001. DETAILED DESCRIPTION

[0070] After long-term and in-depth research, the inventors have prepared a class of compounds that can target ubiquitination and degrade FLT3 protein and its pharmaceutical composition. The compounds can efficiently and selectively degrade FLT3 and other proteins in cells, and therefore have potential uses for treating indications related to abnormal expression of FLT3 kinase. Based on the above findings, the inventors have completed the present invention.

[0071] the term

[0072] In the compounds described herein, when any variable (e.g., R1, R2, etc.) occurs more than once in any component, its definition at each occurrence is independent of the definition at every other occurrence. Likewise, combinations of substituents and variables are permitted as long as such combinations render the compound stable. Lines drawn from substituents into the ring system indicate that the indicated bond may be attached to any substitutable ring atom. If the ring system is polycyclic, it means that such bonds are attached only to any appropriate carbon atom of the adjacent ring. It is to be understood that one of ordinary skill in the art may select substituents and substitution patterns for the compounds of the invention to provide chemically stable compounds that can be readily synthesized from readily available raw materials by techniques in the art and the methods set forth below. If a substituent is itself substituted with more than one group, it is to be understood that these groups may be on the same carbon atom or on different carbon atoms as long as the structure is stable.

[0073] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. The terms used in the description are for the purpose of describing particular embodiments only and are not intended to limit the invention.

[0074] The following terms are used to describe the present invention. Where a term is not specifically defined herein, it is given the art-recognized meaning by one of ordinary skill in the art in which the term is applied in the context of its use to describe the present invention.

[0075] The term "ubiquitin ligase" as used herein refers to a protein family that promotes the transfer of ubiquitin to a specific substrate protein, thereby targeting the substrate protein for degradation. For example, Von Hippel-Lindau E3 ubiquitin ligase or CRBN E3 ubiquitin ligase is such a protein: it alone or in combination with E2 ubiquitin-conjugating enzymes causes the connection of ubiquitin to lysine on the target protein, and then targets a specific protein substrate for degradation by the proteasome. Thus, the E3 ubiquitin ligase that forms a complex alone or with the E2 ubiquitin conjugating enzyme is responsible for the transfer of ubiquitin to the target protein. In general, the ubiquitin ligase participates in polyubiquitination, so that the second ubiquitin is connected to the first ubiquitin; the third ubiquitin is connected to the second ubiquitin, and so on. Polyubiquitination marks proteins for degradation by the proteasome. However, there are some ubiquitination events limited to monoubiquitination, in which only a single ubiquitin is added to the substrate molecule by the ubiquitin ligase. Monoubiquitinated proteins are not targeted to the proteasome for degradation, but instead may be altered in their cellular location or function, for example, by binding to other proteins with domains capable of binding ubiquitin. Further complicating the situation is that different lysines on ubiquitin can be targeted by E3 to form chains. The most common lysine is Lys48 on the ubiquitin chain. This is the lysine used to make polyubiquitin recognized by the proteasome.

[0076] As used herein, the term "alkyl" is intended to include branched and straight chain saturated aliphatic hydrocarbon groups having a specific number of carbon atoms. For example, the definition of "C1-C6" in "C1-C6 alkyl" includes groups having 1, 2, 3, 4, 5 or 6 carbon atoms in a straight or branched arrangement. For example, "C1-C6 alkyl" specifically includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl, and hexyl.

[0077] The term "cycloalkyl" as used herein refers to a saturated or partially unsaturated monocyclic, bicyclic or polycyclic hydrocarbon group whose ring atoms are composed of carbon atoms, and the bicyclic or polycyclic rings include spirocyclic, fused and bridged rings. For example, "cycloalkyl" includes but is not limited to the following groups: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, wait.

[0078] The term "alkoxy" used herein refers to a group having an -O-alkyl structure, such as -OCH3, -OCH2CH3, -OCH2CH2CH3, -O-CH2CH(CH3)2, -OCH2CH2CH2CH3, -O-CH(CH3)2, etc.

[0079] As used herein, the term "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic, bicyclic or polycyclic cyclic substituent, wherein one or more ring atoms are selected from N, O or S(O)m (wherein m is an integer from 0 to 2) heteroatoms, and the remaining ring atoms are carbon, and the bicyclic or polycyclic rings include spirocyclic, fused and bridged rings. For example: morpholinyl, piperidinyl, tetrahydropyrrolyl, pyrrolidinyl, dihydroimidazolyl, dihydroisoxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl, dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl, dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl, tetrahydrofuranyl, tetrahydrothienyl, etc., and N-oxides thereof. Attachment of the heterocyclic substituents may be via a carbon atom or via a heteroatom.

[0080] As used herein, the term "halo" or "halo" refers to chlorine, fluorine, bromine and iodine.

[0081] The present invention provides a compound having a structure shown in formula (I):

[0082]

[0083] in:

[0084] Y is selected from: -CH2-, -C(=O)-;

[0085] R1 and R2 are each independently selected from the following group: -H, -F;

[0086] L has the structure shown below: -Z-(L1) m -;

[0087] wherein m is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30;

[0088] The Z is selected from the following group: a 3-12 membered ring having 0-4 heteroatoms, -NH-, -O-, -C(O)-, -CONH-, -NHCO-, -S-, -CH2-, -C≡C-, -CH=CH-, -P=O-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-;

[0089] Each L1 is independently selected from the following group: a 3-12 membered ring having 0-4 heteroatoms, -NH-, -O-, -C(O)-, -CONH-, -NHCO-, -S-, -CH2-, -C≡C-, -CH=CH-, -P=O-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-;

[0090] The L segment may be optionally replaced by one or more R L substituted, and the R L The substituent is selected from the following group: halogen, -OH, -CN, -CF3, -NH2, -COOH, oxygen atom (=O), substituted or unsubstituted C1-C6 alkyl or haloalkyl, substituted or unsubstituted C1-C6 alkoxy, or 2 R L The radicals and the atoms to which they are attached together form a 3-8 membered ring containing 0-4 heteroatoms;

[0091] The above substitution refers to that one or more H atoms on the group are replaced by a substituent selected from the following group: halogen, deuterium atom.

[0092] In a preferred embodiment of the present invention, the compound is selected from the following group:

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101] Pharmaceutically acceptable salts

[0102] The present invention includes free forms of compounds of formula I, as well as pharmaceutically acceptable salts and stereoisomers thereof. Some specific exemplary compounds herein are protonated salts of amine compounds. The term "free form" refers to amine compounds in non-salt form. Pharmaceutically acceptable salts included include not only the exemplary salts of specific compounds described herein, but also typical pharmaceutically acceptable salts of free forms of all compounds of formula I. The free form of a specific salt of the compound can be isolated using techniques known in the art. For example, the free form can be regenerated by treating the salt with an appropriate dilute aqueous base solution, such as dilute aqueous NaOH, dilute aqueous potassium carbonate, dilute aqueous ammonia, and dilute aqueous sodium bicarbonate. The free form differs somewhat from its respective salt form in certain physical properties, such as solubility in polar solvents, but for the purposes of the invention, such acid and base salts are equivalent to their respective free forms in other pharmaceutical aspects.

[0103] Pharmaceutically acceptable salts of the present invention can be synthesized from compounds of the present invention containing a basic or acidic moiety by conventional chemical methods. Typically, salts of basic compounds are prepared by ion exchange chromatography or by reacting a free base with a stoichiometric amount or excess of an inorganic or organic acid in the desired salt form in an appropriate solvent or combination of solvents. Similarly, salts of acidic compounds are formed by reacting with an appropriate inorganic or organic base.

[0104] Therefore, the pharmaceutically acceptable salts of the compounds of the present invention include inorganic or organic acid salts formed by the basic compounds of the present invention and inorganic or organic acids.

[0105] Wherein, the inorganic acid salt is hydrochloride, sulfate, bisulfate, nitrate, hydrobromide, hydroiodide, carbonate, bicarbonate, sulfite, bisulfite, pyrosulfate, monohydrogen phosphate, dihydrogen phosphate, perchlorate, persulfate, hemisulfate, bisulfate, thiocyanate, phosphate, pyrophosphate, metaphosphate or any combination thereof;

[0106] The organic acid salt is formate, acetate, propionate, butyrate, benzoate, malonate, succinate, pyruvate, methanesulfonate, ethanesulfonate, propanesulfonate, citrate, 4-nitrobenzoate, benzenesulfonate, p-toluenesulfonate, malate, propiolate, 2-butynoate, 2-hydroxy-ethanesulfonate, vinyl acetate, tartrate, L-tartrate, fumarate, isethionate, maleate, lactate, lactobionate, pamoate, salicylate, galactarate, glucoheptonate, mandelate, 1,2-ethanedisulfonate, naphthalenesulfonate , oxalate, trifluoroacetate, trifluoromethanesulfonate, adipate, suberate, sebacate, butyne-1,4-dioate, hexyne-1,6-dioate, glycolate, alginate, ascorbate, isoascorbate, aspartate, L-aspartate, glutamate, L-glutamate, 2-phenoxybenzoate, 2-(4-hydroxybenzoyl)benzoate, acetoacetate, 2-hydroxyethanesulfonate, benzenesulfonate, borate, chlorobenzoate, camphorate, itaconate, camphorsulfonate, L-camphorsulfonate, methylbenzoate, dinitrobenzoate, amino Sulfonate, lactobionate, galacturonate, cyclopentylpropionate, dodecyl sulfate, acrylate, cyclopentanepropionate, glycerophosphate, methoxybenzoate, digluconate, gluconate, heptanoate, hexanoate, 2-hydroxy-ethanesulfonate, trimethylacetate, glucuronate, laurate, phthalate, phenylacetate, lauryl sulfate, 2-acetoxybenzoate, nicotinate, cinnamate, oleate, palmitate, pamoate, pectinate, phthalate, glutarate, hydroxymaleate, hydroxybenzoate, 3-hydroxy-2-naphthoate, 3 -phenylpropionate, isobutyrate, pivalate, picrate, stearate, 2,2-dichloroacetate, acylated amino acid salts, alginate, 4-acetamidobenzenesulfonate, caprate, cholate, octanoate, nonanoate, cyclamate, phthalate, cysteine ​​hydrochloride, sorbate, pamoate, mucate, glycinate hydrochloride, naphthalenedisulfonate, xylenesulfonate, cystine dihydrochloride, undecanoate, polyvinylsulfonate, sulfosalicylate, phenylbutyrate, 4-hydroxybutyrate, polyvinyl sulfate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, valerate, or any combination thereof.

[0107] If the compound of the present invention is acidic, then suitable "pharmaceutically acceptable salts" refer to salts prepared from pharmaceutically acceptable non-toxic bases including inorganic bases and organic bases. Salts derived from inorganic bases include aluminum salts, ammonium salts, calcium salts, copper salts, iron salts, ferrous salts, lithium salts, magnesium salts, manganic salts, manganous salts, potassium salts, sodium salts, zinc salts, etc. Ammonium salts, calcium salts, magnesium salts, potassium salts and sodium salts are particularly preferred. Salts derived from pharmaceutically acceptable organic non-toxic bases, including salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydroxocobalamin, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, guazine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like.

[0108] Berg et al., "Pharmaceutical Salts," J. Pharm. Sci. '1977: 66: 1-19, describes in more detail the preparation of the above-mentioned pharmaceutically acceptable salts and other typical pharmaceutically acceptable salts.

[0109] Drug metabolites and prodrugs

[0110] The metabolites of the compounds and pharmaceutically acceptable salts thereof involved in the present invention, as well as prodrugs that can be converted into the structures of the compounds and pharmaceutically acceptable salts involved in the present application in vivo, are also included in the claims of the present invention.

[0111] Pharmaceutical composition and use thereof

[0112] The present invention also provides a pharmaceutical composition, which comprises an active ingredient within a safe and effective amount, and a pharmaceutically acceptable carrier or excipient. Since the active ingredient of the present application, the compound of formula I and its pharmaceutically acceptable salt have a good inhibitory effect on the abnormal expression of proteins such as FLT3, the pharmaceutical composition can be used to treat tumors, cardiovascular diseases, diabetes, hypertension, muscular dystrophy, Parkinson's disease, Alzheimer's disease and other diseases related to abnormal expression of proteins such as FLT3 in humans or other mammals.

[0113] In one embodiment, the active ingredient of the present application, or a pharmaceutical composition containing the active ingredient can be used to prevent and / or treat tumors such as non-small cell lung cancer, malignant melanoma, prostate cancer, kidney cancer, bladder cancer, ovarian cancer, colon cancer, rectal cancer, breast cancer, cervical cancer, lung cancer, laryngeal cancer, nasopharyngeal cancer, pancreatic cancer, multiple myeloma, B lymphoma, leukemia, etc., or to prevent tumor recurrence after surgery.

[0114] The "active ingredient" described in the present invention refers to the compound of formula I described in the present invention or its pharmaceutically acceptable salt or its stereoisomer or its prodrug molecule.

[0115] The "active ingredients" and pharmaceutical compositions described in the present invention can be used as protein degraders such as FLT3, and can be used to prepare drugs for preventing and / or treating tumors, cardiovascular diseases, diabetes, hypertension, muscular dystrophy, Parkinson's disease, Alzheimer's disease, etc.

[0116] "Safe and effective amount" means: the amount of active ingredient is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of active ingredient per dose, more preferably, 10-200 mg of active ingredient per dose. Preferably, the "one dose" is one tablet.

[0117] "Pharmaceutically acceptable carrier or excipient" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity.

[0118] "Compatibility" herein means that the components in the composition can be blended with the active ingredient of the present invention and with each other without significantly reducing the efficacy of the active ingredient.

[0119] Examples of pharmaceutically acceptable carriers or excipients include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers. Wetting agents (such as sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, or a combination thereof.

[0120] In another preferred embodiment, the compound of formula I of the present invention can form a complex with a macromolecular compound or a polymer through a non-bonding reaction. In another preferred embodiment, the compound of formula I of the present invention as a small molecule can also be connected to a macromolecular compound or a polymer through a chemical bond. The macromolecular compound can be a biological macromolecule such as a high polysaccharide, protein, nucleic acid, polypeptide, etc.

[0121] There is no particular limitation on the administration of the active ingredient or pharmaceutical composition of the present invention. Representative administration methods include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), etc.

[0122] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules.

[0123] In these solid dosage forms, the active ingredient is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients:

[0124] (a) fillers or extenders, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid;

[0125] (b) binders, for example, hydroxymethylcellulose, alginate, gelatin, polyvinyl pyrrolidone, sucrose and gum arabic;

[0126] (c) humectants, for example, glycerin;

[0127] (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate;

[0128] (e) a buffering solvent, such as paraffin;

[0129] (f) absorption accelerators, for example, quaternary ammonium compounds;

[0130] (g) wetting agents, such as cetyl alcohol and glyceryl monostearate;

[0131] (h) adsorbents, for example, kaolin; and

[0132] (i) Lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain a buffering agent.

[0133] The solid dosage forms can also be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They can contain opacifying agents, and the release of the active ingredient in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes.

[0134] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active ingredient, the liquid dosage form may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances. In addition to these inert diluents, the composition may also contain adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and spices.

[0135] In addition to the active ingredients, suspensions may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances.

[0136] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0137] The compounds of the present invention may be administered alone or in combination with other therapeutic agents.

[0138] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 20 to 500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

[0139] Combination medication

[0140] The compound of formula I can be used in combination with other drugs known to treat or improve similar conditions. When administered in combination, the administration method and dosage of the original drug remain unchanged, and the compound of formula I is taken simultaneously or subsequently. When the compound of formula I is taken simultaneously with one or more other drugs, it is preferred to use a pharmaceutical composition containing one or more known drugs and the compound of formula I. Drug combination also includes taking the compound of formula I and one or more other known drugs in overlapping time periods. When the compound of formula I is used in combination with one or more other drugs, the dosage of the compound of formula I or the known drug may be lower than when they are used alone.

[0141] The drugs or active ingredients that can be used in combination with the compounds of formula I include, but are not limited to, estrogen receptor modulators, androgen receptor modulators, retinal receptor modulators, cytotoxins / cytostatics, antiproliferative agents, protein transferase inhibitors, HMG-CoA reductase inhibitors, HIV protein kinase inhibitors, reverse transcriptase inhibitors, angiogenesis inhibitors, cell proliferation and survival signal inhibitors, drugs that interfere with cell cycle checkpoints and apoptosis inducers, cytotoxic drugs, tyrosine protein inhibitors, EGFR inhibitors, VEGFR inhibitors, serine / threonine protein inhibitors, Bcr -Abl inhibitors, c-Kit inhibitors, Met inhibitors, Raf inhibitors, MEK inhibitors, MMP inhibitors, topoisomerase inhibitors, histidine deacetylase inhibitors, proteasome inhibitors, CDK inhibitors, Bcl-2 family protein inhibitors, MDM2 family protein inhibitors, IAP family protein inhibitors, STAT family protein inhibitors, PI3K inhibitors, AKT inhibitors, integrin blockers, interferon-α, interleukin-12, COX-2 inhibitors, p53, p53 activators, VEGF antibodies, EGF antibodies, JAK inhibitors, etc.

[0142] In one embodiment, the drugs or active ingredients that can be used in combination with the compound of formula I include but are not limited to: aldesleukin, alendronic acid, interferon, atrenoin, allopurinol, allopurinol sodium, palonosetron hydrochloride, hexamethylmelamine, aminoglutethimide, amifostine, amrubicin, anacrine, anastrozole, dolasetron, aranesp, arglabin, arsenic trioxide, arnosin, 5-azacytidine, azathioprine, bacillus Calmette-Guérin or tice bacillus Calmette-Guérin, betadine, betamethasone acetate, betamethasone sodium phosphate preparation, bexarotene, bleomycin sulfate, bromourea, bortezomib, busulfan, calcitonin, alezosumab injection, capecitabine, carboplatin, casodex, cefone , cisplatin, cladribine, cladribine, clodrophosphamide, cytarabine, dacarbazine, actinomycin D, daunorubicin liposome, dexamethasone, dexamethasone phosphate, estradiol valerate, denileukin 2, depomet, deslorelin, delazosen, diethylstilbestrol, diflucan, docetaxel, doxorubicin, dronabinol, chin-166-chitosan complex, eligard, rasburicase, epirubicin hydrochloride, aprepitant, epirubicin, epoetin alfa, erythropoietin, epoetin, levamisole tablets, estradiol preparations, 17-β-estradiol, estramustine sodium phosphate, ethinyl estradiol, amifostine, hydroxyphosphoric acid, vanbifu, etoposide, fadrozole, tamoxifen Oxifen preparations, filgrastim, finasteride, firaprestin, floxuridine, fluconazole, fludarabine, 5-fluorodeoxyuridine nucleoside monophosphate, 5-fluorouracil, fluoxymesterone, flutamide, formestan, 1-β-D-arabinofuranosylcytosine-5'-stearoyl phosphate, fotemustine, fulvestrant, gamma globulin, gemcitabine, gemtuzumab, imatinib mesylate, carmustine rice paper capsules, goserelin, granisiron hydrochloride, histrelin, and mexin, hydrocortisone, erythro-hydroxynonyl adenine, hydroxyurea, tentan isobetamin, idarubicin, ifosfamide, interferon α, interferon-α2, interferon α-2A, interferon α-2B, interferon α-nl, interferon α-n3, interferon β , interferon gamma-la, interleukin-2, intron A, Iressa, irinotecan, Kateri, lentinan sulfate, letrozole, leucovorin, leuprorelin acetate, levomizole, levofolinate calcium salt, levothyroxine sodium, levothyroxine sodium preparations, lomustine, lonidamine, dronabinol, nitrogen mustard, methylcobalamin, medroxyprogesterone acetate, megestrol acetate, melphalan, esterified estrogen, 6-mercaptopurine, mesna, methotrexate, methyl aminolevulinate, miltefosine, minocycline, mitomycin C, mitotane, mitoxantrone, trilostane, liposomal doxorubicin citrate, nedaplatin, pegfilgrastim, oprelleukin, neupogen, nilutamide, tamoxifen,NSC-631570, recombinant human interleukin 1-β, octreotide, ondansetron hydrochloride, dehydrocortisone oral solution, oxaliplatin, paclitaxel, prednisone sodium phosphate preparation, pegaspargase, PEGASYS, pentostatin, lysin, pilocarpine hydrochloride, pilocarpine, plicamycin, porfimer sodium, prednimustine, stiprenisolone, prednisone, premarin, procarbazine, recombinant human erythropoietin, raltitrexed, libi, etidronate rhenium-186, rituximab, doxycycline-A, romotide, pilocarpine hydrochloride tablets, octreotide, sarmustine, semustine, sizoran, sobuzosine, sodium methylprednisolone, pafosic acid, stem cell therapy, streptozocin, strontium-89 chloride, levothyroxine sodium, tamoxifen Fen, tamsulosin, tasofenamin, tastolactone, taxotere, tesifazines, temozolomide, teniposide, testosterone propionate, methyltestosterone, thioguanine, thiotepa, thyroid stimulating hormone, tiludronic acid, topotecan, toremifene, tositumomab, trastuzumab, treosulfan, retinoic acid, methotrexate tablets, trimethylmelamine, trimetrexate, triptorelin acetate, triptorelin pamoate, eufodine, uridine, valrubicin, vesirinone, vinblastine, vincristine, vinamide, vinorelbine, virulizine, dexrazoxane, statins ester, zofran, paclitaxel protein stabilized preparation, acolbifene, interferon r-lb, affinitak, aminopterin, arzoxifene, as oprisnil, atamestane, atrasentan, BAY43-9006, Avastin, CCI-779, CDC-501, Celebrex, cetuximab, clenatol, cyproterone acetate, decitabine, DN-101, doxorubicin-MTC, dSLIM, dutasteride, edotecarin, eflornithine, exitecan, fenretinide, histamine dihydrochloride, histrelin hydrogel implant, holmium-166DOTMP, ibandronic acid, interferon gamma, intron-PEG, ixabepilone, keyhole limpet hemocyanin, L-651582, lanreotide, lasofoxifene, libra, lonafamib, miproxifene, minocycline, MS-2 09. Liposomal MTP-PE, MX-6, nafarelin, nemorubicin, nevastatin, noratript, oblimersen, onco-TCS, osidem, paclitaxel polyglutamate, sodium bromoglutamate, PN-401, QS-21, quasi-sea, R-1549, raloxifene, ranpirnase, 13-cis retinoic acid, satraplatin, ciocalcitol, T-138067, tarceva, paclitaxel docosahexaenoic acid, thymosin αl, gazofurin, tipifarnib, tirapazamine, TLK-286, toremifene, trans-MID-lo7R, valspodar, vapreotide, vatalanib, verteporfin, vinflunine, Z-100 and zolelinic acid or a combination thereof.

[0143] The benefits of the present invention lie in that a PROTAC compound with a novel structure is provided, which can efficiently and highly selectively degrade FLT3 protein in cells, effectively inhibit the growth of various tumor cells, and can be used to prepare anti-tumor drugs.

[0144] General Synthesis Methods

[0145] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are generally performed under conventional conditions such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or under conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0146] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0147] The starting materials in the following examples can be obtained from commercial sources, or prepared by methods known in the art, or prepared according to the methods described herein.

[0148] The structures of the compounds were determined by nuclear magnetic resonance ( 1 H-NMR, 13 C-NMR) and / or mass spectrometry (MS). NMR determination uses a Bruker AV-400 nuclear magnetic resonance instrument, the determination solvent is deuterated chloroform (CDCl3) or deuterated dimethyl sulfoxide (DMSO-d6), and TMS is an internal standard. MS determination uses a LCQAD-40000 mass spectrometer. Column chromatography uses 300-400 mesh silica gel (produced by Qingdao Ocean Chemical Plant).

[0149] abbreviation

[0150] Some abbreviations used in this article have the following meanings:

[0151] BOC, Boc tert-butoxycarbonyl

[0152] General synthetic scheme

[0153] Specific embodiments are further illustrated by the following synthetic schemes and examples, which should not be considered limiting in any way. The experimental procedures used to generate the data are discussed in detail below.

[0154]

[0155] Reagents and conditions: (a) N, N-diisopropylethylamine (DIPEA), anhydrous tetrahydrofuran (THF), 50°C, 6h, 68%; (b) glacial acetic acid, sodium triacetoxyborohydride (STAB), anhydrous dichloromethane (DCM), 0°C, overnight reaction, 67%; (c) 10% palladium on carbon, hydrogen, methanol (MeOH), room temperature, overnight reaction, 94%; (d) DIPEA, anhydrous THF, 50°C, 6h, 65%; (e) 10% palladium on carbon, hydrogen, MeOH , room temperature, overnight reaction, 89%; (f) palladium acetate, 4,5-bis(diphenylphosphino-9,9-dimethylxanthene), potassium carbonate, argon protection, anhydrous 1,4-dioxane, 90°C, overnight reaction, 50%; (g) 4N sodium hydroxide, dimethyl sulfoxide / ethanol (DMSO / EtOH=1:2), 30% hydrogen peroxide (H2O2), warming from 0°C to room temperature, 2h, 97%; (h) trifluoroacetic acid (TFA), DCM, room temperature, 2h, 92%.

[0156]

[0157] Reaction reagents and conditions: (a) potassium acetate, glacial acetic acid, 120°C, overnight reaction; (b) 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), DIPEA, N,N-dimethylformamide (DMF), room temperature, 6h, 40%; (c) DIPEA, N-methylpyrrolidone (NMP), 140°C, 6h, 42-62%.

[0158] Example 1: Preparation of 3-((4-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-carbonyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-6a)

[0159] Step 1: 3-Bromo-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carbonitrile (2-12)

[0160]

[0161] 3-Bromo-5-chloro-6-ethylpyrazine-2-carbonitrile 2-11 (4.9 g, 20 mmol), 4-aminotetrahydropyran (2.4 g, 24 mmol) and N,N-diisopropylethylamine (DIPEA, 6.5 g, 50 mmol) were dissolved in anhydrous tetrahydrofuran (THF, 100 mL), reacted at 50 ° C for 6 hours, and after the reaction was completed as monitored by thin layer chromatography (TLC), 200 mL of ice water was slowly added to the system, and solid precipitated. It was filtered under reduced pressure, the filter cake was washed with ether, and vacuum dried to obtain a white solid compound 2-12 (4.2 g, 68%). 1 H NMR(400MHz, CDCl3) δ4.90(d,J=7.5Hz,1H),4.30–4.16(m,1H),4.02(ddd,J=12.1,4.3,2.1Hz,2H),3.56(td,J=11.8,2.2Hz,2H) ,2.55(q,J=7.3Hz,2H),2.04(ddd,J=12.3,4.4,2.1Hz,2H),1.63–1.51(m,2H),1.32(t,J=7.3Hz,3H).MS(ESI),m / z:332.8[M+Na] + .

[0162] Step 2: tert-Butyl 4-(piperidin-4-yl)piperazine-1-carboxylate (2-15)

[0163]

[0164] Benzyl 4-oxopiperidin-1-carboxylate 2-13 (4.7 g, 20 mmol), 1-tert-butyloxycarbonylpiperazine (3.7 g, 20 mmol) and glacial acetic acid (1 mL) were dissolved in anhydrous dichloromethane (DCM, 100 mL), reacted at 0°C for 1 hour, and sodium triacetoxyborohydride (12.7 g, 60 mmol) was slowly added in portions, and stirred at 0°C overnight. After the reaction was completed by TLC monitoring, 200 mL of ice water was slowly added to the system, stirred and quenched, and extracted with dichloromethane three times. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum rotary evaporation. The residue was separated and purified by column chromatography (petroleum ether / ethyl acetate = 5:1) to obtain a white solid 2-14 (5.4 g, 67%). Compound 2-14 (5.4 g, 13.4 mmol) was dissolved in 80 mL of methanol, and an appropriate amount of 10% palladium carbon reagent was added. The reaction system was replaced with hydrogen three times, and stirred at room temperature overnight. After the reaction was completed by TLC monitoring, diatomaceous earth (SiO2) was filtered, and the combined filtrate was evaporated under reduced pressure to remove the solvent to obtain a white solid 2-15 (3.4 g, 94%). MS (ESI), m / z: 270.4 [M+H] + .

[0165] Step 3: tert-Butyl 4-(1-(4-amino-2-methoxyphenyl)piperidin-4-yl)piperazine-1-carboxylate (2-17)

[0166]

[0167] Compound 2-15 (3.4 g, 12.4 mmol), 2-fluoro-5-nitroanisole (2.1 g, 12.4 mmol) and DIPEA (4.0 g, 31 mmol) were dissolved in 80 mL of anhydrous tetrahydrofuran and reacted at 50 °C for 6 hours. After the reaction was completed by TLC monitoring, ice water was added to the system, and ethyl acetate was extracted three times. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum rotary evaporation. The residue was separated and purified by column chromatography (petroleum ether / ethyl acetate = 2:1) to obtain a yellow oily substance 2-16 (3.4 g, 65%). Compound 2-16 (3.4 g, 8.1 mmol) was dissolved in 60 mL of methanol, and an appropriate amount of 10% palladium carbon reagent was added. The reaction system was replaced with hydrogen three times, and stirred at room temperature overnight. After the reaction was completed by TLC monitoring, diatomaceous earth (SiO2) was filtered, and the filtrate was combined and the solvent was removed by vacuum rotary evaporation to obtain a brown solid 2-17 (2.8 g, 89%). 1 H NMR (400MHz, CDCl3) δ6.76 (d, J = 8.1Hz, 1H), 6.28–6.20 (m, 2H), 3.81 (s, 3H), 3.52–3.35 (m, 8H), 2. 58–2.51(m,4H),2.51–2.46(m,2H),2.44–2.37(m,1H),1.86–1.76(m,4H),1.46(s,9H).MS(ESI)for 2-16,m / z:442.9[M+Na] + .MS(ESI)for 2-17,m / z:391.4[M+H] + .

[0168] Step 4: tert-Butyl 4-(1-(4-((3-cyano-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazine-1-carboxylate (2-18)

[0169]

[0170] Compound 2-12 (1.5 g, 5 mmol), compound 2-17 (1.9 g, 5 mmol), palladium acetate (112 mg, 0.5 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethyloxanthene (579 mg, 1 mmol) and potassium carbonate (1.0 g, 7.5 mmol) were dissolved in 30 mL of dry 1,4-dioxane. The reaction system was replaced with argon five times and heated to 90 ° C and stirred overnight. After the reaction was completed by TLC monitoring, the heating was stopped and the system was cooled to room temperature. The solvent was removed by vacuum rotary evaporation, water and ethyl acetate were added and extracted three times, the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered and vacuum rotary evaporation was used to remove the solvent, and the residue was separated and purified by column chromatography (dichloromethane / methanol = 30:1) to obtain a reddish brown solid 2-18 (1.6 g, 50%). 1 HNMR (400MHz, CDCl3) δ7.20 (dd, J=8.6, 2.4Hz, 1H), 6.86 (d, J=8.6Hz, 1H), 6.83 (d, J=2.4Hz ,1H),6.69(s,1H),4.76(d,J=7.2Hz,1H),4.14–4.04(m,1H),4.03–3.96(m,2H),3.87(s,3H) ,3.54–3.42(m,8H),2.60–2.48(m,8H),2.47–2.40(m,1H),2.05–1.97(m,2H),1.92–1.85(m ,2H),1.85–1.74(m,2H),1.61–1.49(m,2H),1.47(s,9H),1.27(t,J=7.4Hz,3H).MS(ESI)for 2-18,m / z:621.4[M+H] + .

[0171] Step 5: tert-Butyl 4-(1-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-ylpiperazine-1-carboxylate (2-19)

[0172]

[0173] Compound 2-18 (1.6 g, 2.5 mmol) and 4N sodium hydroxide solution (4 mL, 16 mmol) were added to a mixed solution of DMSO (10 mL) and ethanol (20 mL), and 30% hydrogen peroxide solution (2 mL, 21 mmol) was slowly added dropwise under ice bath conditions, and the mixture was reacted at room temperature for 2 hours. After the reaction was completed by TLC monitoring, 100 mL of ice water was added to the system, and the mixture was extracted with ethyl acetate three times. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum rotary evaporation to obtain compound 2-19 (1.7 g, 97%). 1 H NMR (400MHz, CDCl3) δ10.72(s,1H),7.50(s,1H),7.46(dd,J=8.6,2.3Hz,1H),6.90(d,J=2.3Hz,1H ),6.84(d,J=8.6Hz,1H),5.17(s,1H),4.61(d,J=7.3Hz,1H),4.26–4.14(m,1H),4.05–3.98(m,2H) ,3.87(s,3H),3.57–3.48(m,4H),3.47–3.42(m,4H),2.60–2.48(m,8H),2.47–2.40(m,1H),2.12–2 .04(m,2H),1.91–1.75(m,4H),1.60–1.51(m,2H),1.46(s,9H),1.29(t,J=7.4Hz,3H).MS(ESI)for 2-19,m / z:639.5[M+H] + .

[0174] Step 6: 6-ethyl-3-((3-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-5)

[0175]

[0176] Compound 2-19 (1.7 g, 2.6 mmol) was dissolved in 10 mL of dichloromethane, and 5 mL of trifluoroacetic acid was added. The mixture was reacted at room temperature for 2 hours. After the reaction was completed by TLC monitoring, the solvent was removed by rotary evaporation under reduced pressure. Saturated sodium bicarbonate solution was slowly added dropwise to the system until the system became weakly alkaline. Dichloromethane was added to dissolve and extracted three times. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. The residue was separated and purified by column chromatography (dichloromethane / methanol = 10:1) to obtain yellow solid compound 2-5 (1.3 g, 92%). 1H NMR (400MHz, DMSO-d6) δ11.01(s,1H),7.53(s,1H),7.26(d,J=8.4Hz,1H),7.23–7.19(m,1H),7. 02(d,J=2.3Hz,1H),6.82(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),4.19–4.06(m,1H),3.96–3.89( m,2H),3.81(s,3H),3.42–3.34(m,6H),2.72–2.65(m,4H),2.58(q,J=7.4Hz,2H),2.49–2.39(m, 5H),2.26–2.17(m,1H),1.89–1.74(m,4H),1.69–1.49(m,4H),1.19(t,J=7.3Hz,3H).MS(ESI)for 2-5,m / z:539.3[M+H] + .

[0177] Step 7: Preparation of 3-((4-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-carbonyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-6a)

[0178]

[0179] Compound 2-5 (135 mg, 0.25 mmol), compound 2-21 (76 mg, 0.25 mmol) and polypeptide condensation agent 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU, 114 mg, 0.3 mmol) were dissolved in 5 mL N,N-dimethylformamide, and N,N-diisopropylethylamine (DIPEA, 131 μL, 0.75 mmol) was added and reacted at room temperature for 6 hours. After the reaction was completed by TLC monitoring, 30 mL of ice water was added to the system, and ethyl acetate was extracted three times. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum rotary evaporation. The residue was separated and purified by column chromatography (dichloromethane / methanol = 20:1) to obtain a light yellow solid compound 2-6a (80 mg, 40%). 1H NMR(500MHz,DMSO-d6)δ11.15(s,1H),11.02(s,1H),8.00(d,J=7.6Hz,1H),7.91(s,1H),7.88(d,J=7.7Hz,1H),7.53(s,1H),7.26(dd,J=8.4,2.3Hz,1H),7.22(d,J=3.2Hz,1H),7.02(d,J=2.3Hz,1H),6.82(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),5.19(dd,J=12.8,5.4Hz,1H),4.15–4.07(m,1H),3.94–3.89(m,2H),3.80(s,3H),3.73–3.59(m,2H),3.41–3.34(m,4H),3.32–3.28(m,2H),2.94–2.85(m,1H),2.67–2.61(m,2H),2.61–2.54(m,4H),2.54–2.51(m,2H),2.49–2.46(m,2H),2.40–2.34(m,1H),2.11–2.04(m,1H),1.88–1.77(m,4H),1.67–1.52(m,4H),1.19(t,J=7.4Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ172.82,169.84,169.67,166.79,166.64,166.60,152.27,151.75,150.15,142.39,136.38,135.38,133.24,131.64,131.59,131.50,123.88,121.64,118.27,111.66,111.53,104.68,66.47,61.15,55.68,50.52,49.13,48.78,48.38,47.59,42.03,32.20,30.95,28.19,24.23,21.97,10.98.HRMS(ESI)for C 42 H 51 O8N 10 [M+H] + ,Calcd:823.3886,Found:823.3872.HPLC analysis:MeCN-H2O(0.1%TFA),RT=8.319min,98.22%purity.

[0180] Example 2: Preparation of 3-((4-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-6b)

[0181]

[0182] Compound 2-5 (162 mg, 0.3 mmol) and 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione 2-22 (124 mg, 0.45 mmol) were dissolved in 5 mL of N-methylpyrrolidone, and N,N-diisopropylethylamine (DIPEA, 149 μL, 0.9 mmol) was added, and heated at 140°C for 6 hours. After the reaction was completed by TLC monitoring, the heating was stopped and the system was cooled to room temperature. 30 mL of ice water was added to the system, and ethyl acetate was extracted three times. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum rotary evaporation. The residue was separated and purified by column chromatography (dichloromethane / methanol = 20:1) to obtain a yellow solid 2-6b (145 mg, 62%). 1 H NMR (500MHz, DMSO-d6) δ11.11(s,1H),11.03(s,1H),7.73–7.67(m,1H),7.53(d,J=3.3Hz,1H),7.38–7.31(m,2H),7.27(dd,J=8.6,2 .3Hz,1H),7.24(d,J=3.3Hz,1H),7.02(d,J=2.3Hz,1H),6.83(d,J=8.6Hz,1H),6.79(d,J=7.5Hz,1H),5.10(dd,J=12.7,5.5Hz,1H), 4.17–4.06(m,1H),3.96–3.89(m,2H),3.82(s,3H),3.41–3.34(m,4H),3.33–3.26(m,4H),2.88(ddd,J=16.7,13.7,5.4Hz,1H),2.80 –2.65(m,4H),2.63–2.51(m,6H),2.39–2.31(m,1H),2.07–2.00(m,1H),1.91–1.81(m,4H),1.69–1.55(m,4H),1.19(t,J=7.3Hz,3H). 13C NMR (126MHz, DMSO-d6) δ172.86,170.06,169.67,167.10,166.34,152.28,151. 75,150.15,149.78,136.45,135.90,135.35,133.70,131.48,123.71,118.27, 116.46,114.79,111.66,111.57,104.76,66.47,61.16,55.72,50.94,50.53,4 8.81,48.66,47.60,32.20,30.98,28.37,24.23,22.09,10.98.HRMS(ESI)forC 41 H 51 N 10 O7[M+H] + , Calcd: 795.3937, Found: 795.3940. HPLC analysis: MeCN-H2O (0.1% TFA), RT=8.654min, 98.73% purity.

[0183] Example 3: Preparation of 3-((4-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-6c)

[0184]

[0185] The synthesis method is the same as Example 2 (yellow solid, 101 mg, 42%). 1HNMR(500MHz,DMSO-d6)δ11.08(s,1H),11.02(s,1H),7.68(d,J=8.5Hz,1H),7.53(d,J=3.4Hz,1H),7.34(d,J=2.2Hz,1H),7.26(ddd,J=8.7,4.6,2.2Hz,2H),7.22(d,J=3.3Hz,1H),7.03(d,J=2.3Hz,1H),6.83(d,J=8.7Hz,1H),6.78(d,J=7.5Hz,1H),5.07(dd,J=12.8,5.4Hz,1H),4.17–4.06(m,1H),3.96–3.89(m,2H),3.82(s,3H),3.44(t,J=4.9Hz,4H),3.36(td,J=12.0,2.1Hz,4H),2.88(ddd,J=16.9,13.8,5.2Hz,1H),2.67(s,4H),2.62–2.54(m,3H),2.53(d,J=13.1Hz,3H),2.41–2.32(m,1H),2.06–1.96(m,1H),1.88–1.82(m,4H),1.62(dtd,J=22.7,11.5,10.7,3.9Hz,4H),1.19(t,J=7.4Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ172.87,170.14,169.70,167.61,167.02,155.27,152.29,151.76,150.17,136.43,135.36,133.85,131.48,124.90,118.28,117.69,111.66,111.57,107.82,104.73,66.49,61.05,55.70,50.48,48.79,48.33,47.62,47.23,32.22,31.02,28.29,24.24,22.22,10.95.HRMS(ESI)for C 41 H 51 N 10 O7[M+H] + ,Calcd:795.3937,Found:795.3941.HPLC analysis:MeCN-H2O(0.1%TFA),RT=8.563min,99.51%purity.

[0186]

[0187] Reaction reagents and conditions: (a) trifluoroacetic anhydride (TFAA), DCM, argon protection, -40°C, 3h; anhydrous tert-butyl alcohol (t-BuOH), room temperature, overnight reaction; (b) potassium bicarbonate, potassium iodide, DMF, 120°C, 6h; (c) trifluoroacetic acid, DCM, room temperature, 2h; (d) HATU, DIPEA, DMF, room temperature, 6h, 37-62%.

[0188] Example 4: Preparation of 3-((4-(4-(4-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-7a)

[0189]

[0190] Compound 2-28a (194 mg, 0.5 mmol) was dissolved in 10 mL of dichloromethane, 2 mL of trifluoroacetic acid was added, and the mixture was reacted at room temperature for 2 hours. After the reaction was completed, the solvent was removed by rotary evaporation under reduced pressure, and the residue was separated and purified by column chromatography (dichloromethane / methanol = 25:1) to obtain yellow solid compound 2-30a. Compound 2-5 (108 mg, 0.2 mmol), compound 2-30a (66 mg, 0.2 mmol) and polypeptide condensation agent HATU (91 mg, 0.24 mmol) were dissolved in 5 mL of N,N-dimethylformamide, DIPEA (105 μL, 0.6 mmol) was added, and the mixture was reacted at room temperature for 6 hours. After the reaction was completed as monitored by TLC, 30 mL of ice water was added to the system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. The residue was separated and purified by column chromatography (dichloromethane / methanol = 15:1) to give a yellow solid compound 2-7a (72 mg, 42%). 1H NMR(500MHz,DMSO-d6)δ11.11(s,1H),11.02(s,1H),7.78(dd,J=8.6,7.3Hz,1H),7.53(d,J=3.3Hz,1H),7.45(d,J=7.2Hz,1H),7.33(d,J=8.6Hz,1H),7.27(dd,J=8.6,2.3Hz,1H),7.22(d,J=3.2Hz,1H),7.02(d,J=2.3Hz,1H),6.83(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),5.18(s,2H),5.11(dd,J=12.7,5.5Hz,1H),4.15–4.07(m,1H),3.95–3.90(m,2H),3.81(s,3H),3.49–3.41(m,4H),3.40–3.33(m,4H),2.89(ddd,J=18.9,13.5,5.4Hz,1H),2.64–2.55(m,5H),2.55–2.51(m,3H),2.49–2.42(m,2H),2.40–2.32(m,1H),2.08–2.01(m,1H),1.88–1.77(m,4H),1.68–1.53(m,4H),1.19(t,J=7.4Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ172.81,169.95,169.66,166.81,165.27,164.87,155.64,152.28,151.74,150.14,136.57,135.41,133.09,131.51,120.21,118.31,116.15,115.46,111.67,111.57,104.74,66.44,66.05,61.21,55.69,54.92,50.47,48.76,48.38,47.58,44.41,41.76,32.19,30.96,28.11,24.21,22.01,10.96.HRMS(ESI)for C 43 H 53 O9N 10 [M+H] + ,Calcd:853.3991,Found:853.3993.HPLC analysis:MeCN-H2O(0.1%TFA),RT=8.466min,97.34%purity.

[0191] Example 5: Preparation of 3-((4-(4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)butyryl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-7b)

[0192]

[0193] The synthesis method is the same as Example 4 (yellow solid, 99 mg, 56%). 1 H NMR(500MHz,DMSO-d6)δ11.10(s,1H),11.01(s,1H),7.81(dd,J=8.5,7.2Hz,1H),7 .56–7.51(m,2H),7.45(d,J=7.2Hz,1H),7.26(dd,J=8.5,2.3Hz,1H),7.21(d,J=3.3 Hz,1H),7.02(d,J=2.3Hz,1H),6.79(dd,J=18.8,8.1Hz,2H),5.09(dd,J=12.7,5.4H z,1H),4.23(t,J=6.3Hz,2H),4.16–4.07(m,1H),3.95–3.89(m,2H),3.81(s,3H),3. 47–3.43(m,4H),3.40–3.34(m,4H),3.32–3.29(m,2H),2.89(ddd,J=16.8,13.7,5.4 Hz,1H),2.62–2.60(m,1H),2.61–2.55(m,2H),2.56–2.51(m,3H),2.49–2.43(m,4H) ,2.30(s,1H),2.07–2.00(m,1H),2.02–1.95(m,2H),1.88–1.82(m,2H),1.76(d,J=1 1.7Hz, 2H), 1.63 (qd, J=12.3, 4.5Hz, 2H), 1.58–1.48 (m, 2H), 1.19 (t, J=7.4Hz, 3H). 13C NMR (126MHz, DMSO-d6) δ172.86,170.03,169.83,169.69,166.90,165.46,155.93,152. 30,151.77,150.17,137.15,136.41,135.39,133.26,131.52,119.85,118.29,116.35,1 15.30,111.67,111.57,104.74,69.82,68.15,66.49,61.17,55.71,49.07,48.78,48.5 6,47.61,45.30,41.46,32.22,30.99,28.14,24.39,24.25,22.06,10.99.HRMS(ESI)for C 45 H 57 O9N 10 [M+H] + , Calcd: 881.4304, Found: 881.4298. HPLC analysis: MeCN-H2O (0.1% TFA), RT=8.701min, 96.82% purity.

[0194] Example 6: Preparation of 3-((4-(4-(4-(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)pentanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-7c)

[0195]

[0196] The synthesis method is the same as Example 4 (yellow solid, 106 mg, 59%). 1H NMR(500MHz,DMSO-d6)δ11.12(s,1H),11.02(s,1H),7.81(dd,J=8.5,7.2Hz,1H),7.53(d,J=3.3Hz,1H),7.51(d,J=8.6Hz,1H),7.44(d,J=7.3Hz,1H),7.26(dd,J=8.6,2.3Hz,1H),7.23(d,J=3.3Hz,1H),7.02(d,J=2.3Hz,1H),6.80(t,J=8.5Hz,2H),5.08(dd,J=12.8,5.5Hz,1H),4.22(t,J=6.2Hz,2H),4.17–4.06(m,1H),3.94–3.89(m,2H),3.80(s,3H),3.46–3.41(m,4H),3.39–3.37(m,2H),3.35–3.27(m,4H),2.88(ddd,J=16.7,13.7,5.4Hz,1H),2.62–2.52(m,4H),2.48–2.46(m,2H),2.45–2.39(m,4H),2.29(tt,J=11.4,3.6Hz,1H),2.06–2.00(m,1H),1.88–1.82(m,2H),1.81–1.73(m,4H),1.72–1.65(m,2H),1.65–1.58(m,2H),1.58–1.49(m,2H),1.18(t,J=7.4Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ172.84,170.35,170.01,169.69,166.92,165.42,155.99,152.29,151.77,150.17,137.10,136.43,135.39,133.27,131.51,119.73,118.29,116.20,115.18,111.67,111.56,104.70,68.76,66.48,61.13,55.69,50.55,49.12,48.78,48.62,47.61,45.32,41.39,32.21,32.01,30.99,28.18,28.17,28.00,24.25,22.08,21.45,10.99.HRMS(ESI)for C 46 H 59 O9N 10 [M+H] +, Calcd: 895.4461, Found: 895.4468. HPLC analysis: MeCN-H2O (0.1% TFA), RT=8.923min, 95.63% purity.

[0197] Example 7: Preparation of 3-((4-(4-(4-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)hexanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-7d)

[0198]

[0199] The synthesis method is the same as Example 4 (yellow solid, 73 mg, 40%). 1 H NMR(500MHz,DMSO-d6)δ11.11(s,1H),11.02(s,1H),7.81(t,J=7.9Hz,1H),7 .52(d,J=8.6Hz,2H),7.44(d,J=7.2Hz,1H),7.26(dd,J=8.6,2.3Hz,1H),7.21 (d,J=3.3Hz,1H),7.02(d,J=2.3Hz,1H),6.81(d,J=8.6Hz,1H),6.78(d,J=7.6 Hz,1H),5.08(dd,J=12.8,5.4Hz,1H),4.20(t,J=6.4Hz,2H),4.16–4.07(m,1H ),3.96–3.88(m,2H),3.81(s,3H),3.46–3.40(m,4H),3.40–3.29(m,4H),2.9 2–2.84(m,1H),2.62–2.55(m,4H),2.54–2.43(m,6H),2.40–2.34(m,1H),2.32 (t,J=7.4Hz,2H),2.06–1.99(m,1H),1.87–1.82(m,2H),1.81–1.74(m,4H),1. 68–1.60(m,2H),1.60–1.52(m,4H),1.51–1.43(m,2H),1.19(t,J=7.4Hz,3H). 13C NMR(126MHz,DMSO-d6)δ172.90,170.42,170.06,169.73,166.93,165.40,156.07,152.31,15 1.80,150.20,137.12,136.44,135.41,133.29,131.55,119.81,118.29,116.24,115.22,111 .68,111.60,104.75,68.77,66.51,61.17,55.72,50.54,49.13,48.79,48.63,47.64,45.38, 41.38,32.29,32.24,31.02,28.33,28.20,25.20,24.57,24.28,22.09,11.00.HRMS(ESI)for C 47 H 61 O9N 10 [M+H] + , Calcd: 909.4618, Found: 909.4625. HPLC analysis: MeCN-H2O (0.1% TFA), RT=9.027min, 95.35% purity.

[0200] Example 8: Preparation of 3-((4-(4-(4-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide Preparation (2-7e)

[0201]

[0202] The synthesis method is the same as Example 4 (yellow solid, 96 mg, 51%). 1H NMR(500MHz,DMSO-d6)δ11.10(s,1H),11.00(s,1H),7.79(dd,J=8.5,7.3Hz,1H),7.52–7.47(m,2H),7.42(d,J=7.2Hz,1H),7.24(dd,J=8.6,2.3Hz,1H),7.20(d,J=3.3Hz,1H),7.00(d,J=2.3Hz,1H),6.79(d,J=8.7Hz,1H),6.77(d,J=7.6Hz,1H),5.06(dd,J=12.8,5.5Hz,1H),4.18(t,J=6.4Hz,2H),4.13–4.05(m,1H),3.93–3.88(m,2H),3.79(s,3H),3.45–3.37(m,4H),3.37–3.33(m,2H),3.31–3.27(m,2H),2.86(ddd,J=16.9,13.8,5.4Hz,1H),2.60–2.50(m,4H),2.47–2.40(m,6H),2.33–2.28(m,1H),2.26(t,J=7.5Hz,2H),2.01(dtd,J=13.0,5.7,5.3,2.1Hz,1H),1.83(dd,J=11.7,4.1Hz,2H),1.79–1.70(m,4H),1.66–1.57(m,2H),1.57–1.50(m,2H),1.49–1.41(m,4H),1.37–1.25(m,4H),1.17(t,J=7.4Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ172.79,170.44,169.97,169.64,166.86,165.32,156.03,152.26,151.73,150.13,137.03,136.41,135.34,133.25,131.47,119.78,118.25,116.21,115.12,111.66,111.56,104.76,68.78,66.43,61.11,55.68,50.49,49.10,48.73,48.60,47.56,45.36,41.35,32.21,32.18,30.96,28.70,28.48,28.38,28.16,25.21,24.77,24.19,22.01,10.95.HRMS(ESI)for C 49 H 65 O9N 10 [M+H] +, Calcd: 937.4930, Found: 937.4914. HPLC analysis: MeCN-H2O (0.1% TFA), RT=9.581min, 96.27% purity.

[0203] Example 9: Preparation of 3-((4-(4-(4-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)decanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-7f)

[0204]

[0205] The synthesis method is the same as Example 4 (yellow solid, 71 mg, 37%). 1 H NMR(500MHz,DMSO-d6)δ11.10(s,1H),11.01(s,1H),7.80(dd,J=8.5,7.3Hz,1H),7.5 5–7.52(m,1H),7.50(d,J=8.6Hz,1H),7.43(d,J=7.2Hz,1H),7.25(dd,J=8.6,2.3Hz, 1H),7.21(d,J=3.3Hz,1H),7.02(d,J=2.3Hz,1H),6.81(d,J=8.6Hz,1H),6.77(d,J=7 .5Hz,1H),5.08(dd,J=12.8,5.4Hz,1H),4.19(t,J=6.4Hz,2H),4.15–4.06(m,1H),3.9 5–3.88(m,2H),3.80(s,3H),3.46–3.39(m,4H),3.39–3.30(m,4H),2.93–2.82(m,1H) ,2.62–2.54(m,4H),2.54–2.42(m,6H),2.36–2.29(m,1H),2.26(t,J=7.5Hz,2H),2.05 –1.99(m,1H),1.88–1.82(m,2H),1.81–1.71(m,4H),1.68–1.58(m,2H),1.58–1.51(m ,2H),1.51–1.41(m,4H),1.37–1.31(m,2H),1.30–1.25(m,6H),1.19(t,J=7.3Hz,3H). 13CNMR(151MHz,DMSO-d6)δ172.79,170.44,169.96,169.64,166.86,165.31,156.03,152.26,151.73, 150.13,137.03,136.41,135.34,133.25,131.47,119.78,118.25,116.22,115.12,111.66,111.57, 104.76,68.79,66.43,61.11,55.68,50.50,49.10,48.73,48.60,47.57,45.38,41.36,32.23,32.18 ,30.96,28.88,28.77,28.75,28.61,28.41,28.16,25.25,24.85,24.20,22.00,10.95.HRMS(ESI)for C 51 H 69 O9N 10 [M+H] + , Calcd: 965.5244, Found: 965.5247. HPLC analysis: MeCN-H2O (0.1% TFA), RT=10.209min, 95.42% purity.

[0206] Example 10: Preparation of 3-((4-(4-(4-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)acetyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-8a)

[0207]

[0208] The synthesis method is the same as Example 4 (yellow solid, 75 mg, 44%). 1H NMR(500MHz,DMSO-d6)δ11.11(s,1H),11.02(s,1H),7.83(d,J=8.3Hz,1H),7.53(d,J=3.3Hz,1H),7.43(d,J=2.3Hz,1H),7.34(dd,J=8.3,2.3Hz,1H),7.27(dd,J=8.6,2.3Hz,1H),7.22(d,J=3.2Hz,1H),7.02(d,J=2.3Hz,1H),6.82(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),5.16–5.08(m,3H),4.16–4.06(m,1H),3.95–3.89(m,2H),3.81(s,3H),3.49–3.42(m,4H),3.41–3.34(m,4H),2.93–2.85(m,1H),2.63–2.53(m,6H),2.53–2.47(m,4H),2.39–2.32(m,1H),2.08–2.02(m,1H),1.88–1.77(m,4H),1.69–1.53(m,4H),1.19(t,J=7.4Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ172.88,170.03,169.72,167.01,166.86,164.92,163.76,152.30,151.79,150.19,136.43,135.41,133.74,131.54,125.20,123.23,121.23,118.30,111.68,111.59,109.13,104.73,66.51,66.13,61.20,55.72,50.56,49.03,48.83,48.52,47.63,44.46,41.77,32.23,31.01,28.20,24.27,22.11,10.99.HRMS(ESI)for C 43 H 53 O9N 10 [M+H] + ,Calcd:853.3991,Found:853.3984.HPLC analysis:MeCN-H2O(0.1%TFA),RT=8.349min,95.10%purity.

[0209] Example 11: Preparation of 3-((4-(4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)butanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-8b)

[0210]

[0211] The synthesis method is the same as Example 4 (yellow solid, 81 mg, 46%). 1 H NMR (500MHz, DMSO-d6) δ11.11(s,1H),11.02(s,1H),7.84(d,J=8.3Hz,1H),7.53 (d,J=3.4Hz,1H),7.44(d,J=2.3Hz,1H),7.36(dd,J=8.3,2.3Hz,1H),7.26(dd,J =8.6,2.3Hz,1H),7.22(d,J=3.2Hz,1H),7.02(d,J=2.3Hz,1H),6.81(d,J=8.6Hz ,1H),6.78(d,J=7.5Hz,1H),5.11(dd,J=12.7,5.4Hz,1H),4.20(t,J=6.5Hz,2H), 4.15–4.06(m,1H),3.94–3.89(m,2H),3.80(s,3H),3.50–3.41(m,4H),3.40–3.2 9(m,4H),2.88(ddd,J=16.7,13.6,5.4Hz,1H),2.62–2.51(m,6H),2.49–2.43(m, 6H),2.36–2.28(m,1H),2.06–2.01(m,1H),2.01–1.95(m,2H),1.87–1.82(m,2H) ,1.81–1.74(m,2H),1.68–1.59(m,2H),1.58–1.50(m,2H),1.19(t,J=7.3Hz,3H). 13C NMR (126MHz, DMSO-d6) δ172.85,170.00,169.79,169.70,166.93,166.86,164.01,152. 29,151.77,150.17,136.42,135.38,133.96,131.51,125.40,122.94,120.68,118.27,1 11.67,111.56,109.01,104.72,68.27,66.49,64.98,61.15,55.70,50.54,48.99,48.5 8,47.61,45.29,41.53,32.22,30.99,28.34,28.18,24.25,22.12,10.98.HRMS(ESI)for C 45 H 57 N 10 O9[M+H] + , Calcd: 881.4304, Found: 881.4307. HPLC analysis: MeCN-H2O (0.1% TFA), RT=8.723min, 95.17% purity.

[0212] Example 12: Preparation of 3-((4-(4-(4-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)hexanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-8c)

[0213]

[0214] The synthesis method is the same as Example 4 (yellow solid, 107 mg, 59%). 1H NMR(500MHz,DMSO-d6)δ11.11(s,1H),11.02(s,1H),7.83(d,J=8.3Hz,1H),7.53(d,J=3.5Hz,1H),7.42(d,J=2.3Hz,1H),7.35(dd,J=8.3,2.3Hz,1H),7.26(dd,J=8.6,2.3Hz,1H),7.22(d,J=3.1Hz,1H),7.02(d,J=2.3Hz,1H),6.81(d,J=8.7Hz,1H),6.78(d,J=7.5Hz,1H),5.11(dd,J=12.8,5.4Hz,1H),4.17(t,J=6.4Hz,2H),4.14–4.08(m,1H),3.95–3.88(m,2H),3.80(s,3H),3.49–3.42(m,4H),3.41–3.35(m,4H),2.88(ddd,J=16.6,13.7,5.5Hz,1H),2.62–2.52(m,6H),2.49–2.46(m,4H),2.41–2.35(m,1H),2.33(t,J=7.4Hz,2H),2.05–1.99(m,1H),1.88–1.82(m,2H),1.82–1.74(m,4H),1.68–1.60(m,2H),1.60–1.52(m,4H),1.48–1.41(m,2H),1.19(t,J=7.4Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ172.85,170.41,170.01,169.70,166.95,166.87,164.15,152.29,151.78,150.17,136.34,135.43,133.98,131.53,125.36,122.89,120.75,118.30,111.68,111.55,108.86,104.71,68.80,66.49,61.25,55.71,50.46,49.08,48.99,48.59,47.62,45.15,41.19,32.22,32.16,31.00,28.29,28.08,25.24,24.54,24.26,22.12,10.98.HRMS(ESI)for C 47 H 61 N 10 O9[M+H] +, Calcd: 909.4618, Found: 909.4616. HPLC analysis: MeCN-H2O (0.1% TFA), RT=9.150min, 98.25% purity.

[0215] Example 13: Preparation of 3-((4-(4-(4-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)octanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-8d)

[0216]

[0217] The synthesis method is the same as Example 4 (yellow solid, 116 mg, 62%). 1 H NMR (500MHz, DMSO-d6) δ11.10(s,1H),11.01(s,1H),7.82(d,J=8.3Hz,1H),7.52(d,J=3 .3Hz,1H),7.42(d,J=2.3Hz,1H),7.34(dd,J=8.3,2.3Hz,1H),7.26(dd,J=8.6,2.3Hz,1H ),7.21(d,J=3.2Hz,1H),7.02(d,J=2.3Hz,1H),6.81(d,J=8.6Hz,1H),6.78(d,J=7.5Hz ,1H),5.11(dd,J=12.8,5.4Hz,1H),4.17(t,J=6.5Hz,2H),4.15–4.06(m,1H),3.94–3.89 (m,2H),3.80(s,3H),3.48–3.39(m,4H),3.39–3.33(m,4H),2.88(ddd,J=16.6,13.7,5. 3Hz,1H),2.62–2.55(m,4H),2.55–2.42(m,6H),2.37–2.31(m,1H),2.28(t,J=7.5Hz,2H) ,2.07–2.00(m,1H),1.88–1.81(m,2H),1.81–1.72(m,4H),1.68–1.60(m,2H),1.60–1.53 (m,2H),1.53–1.46(m,2H),1.46–1.39(m,2H),1.38–1.28(m,4H),1.19(t,J=7.4Hz,3H). 13C NMR (126MHz, DMSO-d6) δ172.83,170.45,169.99,169.68,166.94,166.86,164.14,152.28,151 .76,150.16,136.40,135.38,133.97,131.49,125.34,122.87,120.75,118.26,111.67,111.55 ,108.84,104.71,68.82,66.48,61.15,55.69,50.53,49.13,48.97,48.63,47.60,45.38,41.3 8,32.22,30.99,28.74,28.56,28.37,28.18,25.30,24.81,24.24,22.11,10.98.HRMS(ESI)for C 49 H 65 O9N 10 [M+H] + , Calcd: 937.4931, Found: 937.4938. HPLC analysis: MeCN-H2O (0.1% TFA), RT=9.747min, 97.94% purity.

[0218] Example 14: Preparation of 3-((4-(4-(4-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)decanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-8e)

[0219]

[0220] The synthesis method is the same as Example 4 (yellow solid, 110 mg, 57%). 1H NMR(500MHz,DMSO-d6)δ11.11(s,1H),11.01(s,1H),7.82(d,J=8.3Hz,1H),7.53(d,J=3.3Hz,1H),7.42(d,J=2.3Hz,1H),7.34(dd,J=8.3,2.3Hz,1H),7.26(dd,J=8.6,2.3Hz,1H),7.22(d,J=3.3Hz,1H),7.01(d,J=2.3Hz,1H),6.81(d,J=8.6Hz,1H),6.78(d,J=7.6Hz,1H),5.11(dd,J=12.8,5.4Hz,1H),4.16(t,J=6.5Hz,2H),4.14–4.07(m,1H),3.94–3.89(m,2H),3.80(s,3H),3.48–3.39(m,4H),3.39–3.30(m,4H),2.88(ddd,J=16.8,13.7,5.4Hz,1H),2.62–2.55(m,4H),2.54–2.41(m,6H),2.37–2.29(m,1H),2.27(t,J=7.5Hz,2H),2.07–2.00(m,1H),1.88–1.81(m,2H),1.81–1.70(m,4H),1.68–1.60(m,2H),1.60–1.51(m,2H),1.51–1.44(m,2H),1.44–1.38(m,2H),1.37–1.30(m,2H),1.30–1.25(m,6H),1.19(t,J=7.3Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ172.82,170.44,169.99,169.68,166.93,166.85,164.13,152.27,151.75,150.15,136.40,135.37,133.96,131.48,125.33,122.86,120.73,118.25,111.66,111.54,108.83,104.70,68.82,66.47,61.14,55.69,50.53,49.12,48.97,47.60,45.39,41.38,32.25,32.20,30.98,28.94,28.86,28.80,28.70,28.41,28.17,25.39,24.88,24.24,22.10,10.97.HRMS(ESI)for C 51 H 69 O9N 10[M+H] + , Calcd: 965.5244, Found: 965.5253. HPLC analysis: MeCN-H2O (0.1% TFA), RT=10.518min, 98.85% purity.

[0221]

[0222] Reaction reagents and conditions: (a) dichlorothionyl (SOCl2), room temperature, 2h; anhydrous tert-butanol (t-BuOH), room temperature, overnight reaction; (b) TFAA, DCM, -40℃, 3h; anhydrous tert-butanol (t-BuOH), room temperature, overnight reaction; (c) 10% palladium carbon, hydrogen, MeOH, room temperature, overnight reaction; (d) DIPEA, NMP, 120℃, 5h; (e) TFA, DCM, room temperature, 2h; (f) HATU, DIPEA, DMF, room temperature, 6h, 27-68%.

[0223] Example 15: Preparation of 3-((4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)glycyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-9a)

[0224]

[0225] The synthesis method is the same as Example 4 (yellow solid, 54 mg, 32%). 1H NMR(500MHz,DMSO-d6)δ11.11(s,1H),11.02(s,1H),7.61(t,J=7.8Hz,1H),7.56–7.50(m,1H),7.27(d,J=8.6Hz,1H),7.24–7.21(m,1H),7.13–7.04(m,3H),7.02(t,J=1.9Hz,1H),6.82(d,J=8.6Hz,1H),6.79(d,J=7.4Hz,1H),5.07(dd,J=13.1,5.3Hz,1H),4.19(d,J=4.5Hz,2H),4.16–4.06(m,1H),3.94–3.89(m,2H),3.81(s,3H),3.54–3.46(m,4H),3.40–3.34(m,4H),2.89(ddd,J=18.6,14.2,5.4Hz,1H),2.64–2.54(m,6H),2.54–2.47(m,4H),2.40–2.32(m,1H),2.07–2.00(m,1H),1.88–1.77(m,4H),1.67–1.53(m,4H),1.18(t,J=7.4Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ172.89,170.15,169.68,168.84,167.40,166.21,152.28,151.76,150.16,145.45,136.41,136.22,135.38,132.04,131.51,118.29,111.67,111.56,110.84,109.55,104.71,66.48,61.17,55.69,50.56,48.74,48.60,48.44,47.60,44.21,43.64,42.04,32.21,28.20,24.24,22.18,10.98.HRMS(ESI)for C 43 H 54 O8N 11 [M+H] + ,Calcd:852.4151,Found:852.4155.HPLC analysis:MeCN-H2O(0.1%TFA),RT=8.698min,95.73%purity.

[0226] Example 16: Preparation of 3-((4-(4-(4-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)propanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-9b)

[0227]

[0228] The synthesis method is the same as Example 4 (yellow solid, 83 mg, 48%). 1 H NMR(500MHz,DMSO-d6)δ11.08(s,1H),11.00(s,1H),7.57(dd,J=8.6,7.1Hz,1H) ,7.51(d,J=3.3Hz,1H),7.24(dd,J=8.6,2.3Hz,1H),7.20(d,J=3.3Hz,1H),7.12 (d,J=8.6Hz,1H),7.04–6.98(m,2H),6.79(d,J=8.7Hz,1H),6.78–6.74(m,2H),5 .03(dd,J=12.7,5.4Hz,1H),4.14–4.05(m,1H),3.93–3.86(m,2H),3.79(s,3H),3 .53(q,J=6.4Hz,2H),3.47–3.35(m,4H),3.35–3.25(m,4H),2.86(ddd,J=16.8,1 3.6,5.3Hz,1H),2.63(t,J=6.3Hz,2H),2.59–2.50(m,3H),2.47–2.36(m,7H),2. 32–2.25(m,1H),2.03–1.97(m,1H),1.83(dd,J=12.6,3.8Hz,2H),1.73(d,J=11. 8Hz,2H),1.61(qd,J=12.3,4.4Hz,2H),1.55–1.46(m,2H),1.17(t,J=7.4Hz,3H). 13CNMR(151MHz,DMSO-d6)δ172.80,170.08,169.65,169.04,168.82,167.30,152.27,151.7 4,150.14,146.19,136.41,136.34,135.34,132.24,131.48,118.27,117.14,111.66,111. 58,110.49,109.19,104.76,66.44,61.10,55.69,50.51,48.83,48.54,48.42,47.57,45. 28,41.44,38.20,32.18,31.89,30.98,28.12,28.08,24.20,22.17,10.96.HRMS(ESI)forC 44 H 56 O8N 11 [M+H] + , Calcd: 866.4308, Found: 866.4295. HPLC analysis: MeCN-H2O (0.1% TFA), RT=8.793min, 98.72% purity.

[0229] Example 17: Preparation of 3-((4-(4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-9c)

[0230]

[0231] The synthesis method is the same as Example 4 (yellow solid, 90 mg, 51%). 1H NMR(500MHz,DMSO-d6)δ11.10(s,1H),11.02(s,1H),7.58(dd,J=8.6,7.1Hz,1H),7.53(d,J=3.3Hz,1H),7.26(dd,J=8.5,2.3Hz,1H),7.22(d,J=3.3Hz,1H),7.17(d,J=8.6Hz,1H),7.05–7.00(m,2H),6.81(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),6.65(t,J=6.2Hz,1H),5.06(dd,J=12.7,5.4Hz,1H),4.16–4.07(m,1H),3.95–3.89(m,2H),3.81(s,3H),3.49–3.40(m,4H),3.40–3.33(m,4H),3.32–3.29(m,2H),2.94–2.83(m,1H),2.62–2.53(m,4H),2.52–2.41(m,6H),2.39(t,J=7.0Hz,2H),2.34–2.26(m,1H),2.06–1.99(m,1H),1.88–1.82(m,2H),1.82–1.73(m,4H),1.68–1.58(m,2H),1.58–1.48(m,2H),1.19(t,J=7.4Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ172.89,170.17,170.03,169.69,168.85,167.37,152.28,151.76,150.17,146.43,136.41,136.29,135.38,132.28,131.51,118.28,117.31,111.66,111.56,110.42,109.11,104.72,66.48,61.17,55.70,50.53,48.99,48.55,47.61,45.26,41.61,41.47,32.21,31.02,29.39,28.15,24.28,24.24,22.20,10.99.HRMS(ESI)for C 45 H 58 O8N 11 [M+H] + ,Calcd:880.4464,Found:880.4470.HPLC analysis:MeCN-H2O(0.1%TFA),RT=8.863min,99.50%purity.

[0232] Example 18: Preparation of 3-((4-(4-(4-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-9d)

[0233]

[0234] The synthesis method is the same as Example 4 (yellow solid, 96 mg, 53%). 1 H NMR(600MHz,DMSO-d6)δ11.09(s,1H),11.01(s,1H),7.57(dd,J=8.6,7.1Hz, 1H),7.53(d,J=3.3Hz,1H),7.26(dd,J=8.6,2.4Hz,1H),7.21(d,J=3.3Hz,1H) ,7.09(d,J=8.7Hz,1H),7.04–6.99(m,2H),6.81(d,J=8.6Hz,1H),6.78(d,J= 7.5Hz,1H),6.52(t,J=6.0Hz,1H),5.05(dd,J=12.9,5.5Hz,1H),4.15–4.07(m ,1H),3.94–3.89(m,2H),3.80(s,3H),3.44–3.38(m,4H),3.34–3.24(m,6H), 2.92–2.84(m,1H),2.62–2.51(m,4H),2.49–2.39(m,6H),2.32–2.25(m,3H),2 .06–2.00(m,1H),1.87–1.82(m,2H),1.80–1.73(m,2H),1.67–1.60(m,2H),1. 60–1.55(m,2H),1.55–1.49(m,4H),1.39–1.31(m,2H),1.18(t,J=7.4Hz,3H). 13C NMR(151MHz,DMSO-d6)δ172.85,170.38,170.13,169.68,168.98,167.34,152.29,151.77,15 0.17,146.46,136.45,136.32,135.37,132.22,131.52,118.28,117.21,111.67,111.61,110 .41,109.02,104.79,66.47,61.11,55.71,50.52,49.12,48.63,48.57,47.60,45.39,41.79, 41.41,32.21,32.17,31.01,28.56,28.21,26.11,24.59,24.23,22.20,10.97.HRMS(ESI)for C 47 H 62 O8N 11 [M+H] + , Calcd: 908.4777, Found: 908.4771. HPLC analysis: MeCN-H2O (0.1% TFA), RT=9.386min, 98.15% purity.

[0235] Example 19: Preparation of 3-((4-(4-(4-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-9e)

[0236]

[0237] The synthesis method is the same as Example 4 (yellow solid, 127 mg, 68%). 1H NMR(500MHz,DMSO-d6)δ11.09(s,1H),11.02(s,1H),7.58(dd,J=8.6,7.1Hz,1H),7.52(d,J=3.3Hz,1H),7.26(dd,J=8.6,2.3Hz,1H),7.22(d,J=3.3Hz,1H),7.09(d,J=8.6Hz,1H),7.02(s,1H),7.01(d,J=5.3Hz,1H),6.81(d,J=8.7Hz,1H),6.77(d,J=7.5Hz,1H),6.52(t,J=5.9Hz,1H),5.05(dd,J=12.7,5.5Hz,1H),4.16–4.06(m,1H),3.94–3.89(m,2H),3.80(s,3H),3.47–3.38(m,4H),3.38–3.33(m,2H),3.32–3.25(m,4H),2.88(ddd,J=16.8,13.7,5.4Hz,1H),2.62–2.51(m,4H),2.50–2.40(m,6H),2.35–2.29(m,1H),2.27(t,J=7.5Hz,2H),2.05–1.99(m,1H),1.85(ddd,J=12.3,4.4,1.9Hz,2H),1.81–1.74(m,2H),1.68–1.59(m,2H),1.59–1.51(m,4H),1.51–1.44(m,2H),1.37–1.26(m,6H),1.19(t,J=7.4Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ172.81,170.41,170.10,169.65,168.96,167.31,152.26,151.73,150.13,146.43,136.40,136.28,135.34,132.20,131.47,118.25,117.19,111.66,111.56,110.37,109.01,104.75,66.43,61.11,55.68,50.49,49.09,48.60,48.54,47.57,45.36,41.82,41.36,32.20,32.18,30.98,28.72,28.64,28.57,28.16,26.23,24.77,24.20,22.15,10.94.HRMS(ESI)for C 49 H 66 O8N 11 [M+H] +, Calcd: 936.5090, Found: 936.5074. HPLC analysis: MeCN-H2O (0.1% TFA), RT=10.000min, 96.27% purity.

[0238] Example 20: Preparation of 3-((4-(4-(4-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)decanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-9f)

[0239]

[0240] The synthesis method is the same as Example 4 (yellow solid, 81 mg, 42%). 1 H NMR(500MHz,DMSO-d6)δ11.10(s,1H),11.02(s,1H),7.57(t,J=7.8Hz,1H),7.53 (d,J=3.3Hz,1H),7.26(dd,J=8.5,2.3Hz,1H),7.22(d,J=3.3Hz,1H),7.08(d,J=8 .6Hz,1H),7.04–6.99(m,2H),6.84–6.76(m,2H),6.52(t,J=5.9Hz,1H),5.05(dd ,J=12.7,5.4Hz,1H),4.16–4.06(m,1H),3.95–3.88(m,2H),3.80(s,3H),3.46–3. 39(m,4H),3.39–3.34(m,2H),3.33–3.24(m,4H),2.93–2.82(m,1H),2.62–2.54( m,4H),2.53–2.42(m,6H),2.35–2.29(m,1H),2.26(t,J=7.5Hz,2H),2.04–1.99(m ,1H),1.87–1.81(m,2H),1.81–1.75(m,2H),1.68–1.60(m,2H),1.60–1.52(m,4H) ,1.50–1.42(m,2H),1.37–1.28(m,4H),1.28–1.24(m,6H),1.18(t,J=7.3Hz,3H). 13C NMR(126MHz,DMSO-d6)δ172.87,170.47,170.16,169.68,168.99,167.34,152.28,151.75,150.16,1 46.45,136.39,136.32,135.38,132.22,131.50,118.27,117.21,111.66,111.54,110.40,109.01,1 04.71,66.47,61.17,55.69,50.51,49.11,48.62,48.55,47.60,45.35,41.84,41.34,32.24,32.20, 31.01,28.95,28.85,28.79,28.74,28.69,28.14,26.33,24.87,24.23,22.18,10.98.HRMS(ESI)for C 51 H 70 O8N 11 [M+H] + , Calcd: 964.5403, Found: 964.5395. HPLC analysis: MeCN-H2O (0.1% TFA), RT=10.703min, 96.88% purity.

[0241] Example 21: Preparation of 3-((4-(4-(4-(12-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)dodecanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-9 g)

[0242]

[0243] The synthesis method is the same as Example 4 (yellow solid, 60 mg, 30%). 1H NMR(500MHz,DMSO-d6)δ11.09(s,1H),11.01(s,1H),7.57(dd,J=8.6,7.1Hz,1H),7.52(d,J=3.3Hz,1H),7.26(dd,J=8.6,2.3Hz,1H),7.21(d,J=2.8Hz,1H),7.08(d,J=8.6Hz,1H),7.04–6.99(m,2H),6.81(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),6.54–6.49(m,1H),5.04(dd,J=12.7,5.5Hz,1H),4.17–4.06(m,1H),3.95–3.88(m,2H),3.80(s,3H),3.49–3.39(m,4H),3.39–3.34(m,2H),3.32–3.24(m,4H),2.88(ddd,J=16.9,13.7,5.5Hz,1H),2.62–2.55(m,3H),2.54–2.51(m,2H),2.49–2.40(m,5H),2.35–2.29(m,1H),2.26(t,J=7.5Hz,2H),2.05–1.99(m,1H),1.87–1.81(m,2H),1.81–1.74(m,2H),1.68–1.60(m,2H),1.60–1.51(m,4H),1.50–1.43(m,2H),1.37–1.28(m,4H),1.28–1.23(m,10H),1.19(t,J=7.3Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ172.86,170.45,170.15,169.68,168.98,167.34,152.27,151.75,150.16,146.44,136.39,136.30,135.38,132.21,131.48,118.26,117.19,111.67,111.54,110.39,109.00,104.71,66.47,61.15,55.69,50.52,49.11,48.63,48.55,47.59,45.38,41.85,41.36,32.24,32.20,31.01,29.03,28.99,28.92,28.80,28.69,28.16,26.35,24.88,24.23,22.18,10.97.HRMS(ESI)for C 53 H 74 O8N 11 [M+H]+ , Calcd: 992.5716, Found: 992.5724. HPLC analysis: MeCN-H2O (0.1% TFA), RT=11.528min, 98.15% purity.

[0244] Example 22: Preparation of 3-((4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)glycyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-10a)

[0245]

[0246] The synthesis method is the same as Example 4 (yellow solid, 56 mg, 33%). 1 H NMR (500MHz, DMSO-d6) δ11.05(s,1H),11.02(s,1H),7.57(d,J=8.4Hz,1H),7.53(d,J=3.3Hz,1H),7.27(dd,J=8.6,2.3Hz,1H),7.22(d,J= 3.3Hz,1H),7.12(s,1H),7.09(t,J=5.1Hz,1H),7.04–6.99(m,2H),6.82(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),5.04(dd,J=12.8,5.4Hz,1 H),4.17–4.06(m,3H),3.95–3.89(m,2H),3.81(s,3H),3.51(q,J=5.5,4.6Hz,4H),3.40–3.34(m,4H),2.88(ddd,J=17.1,14.1,5.6Hz,1H) ,2.61–2.55(m,5H),2.54–2.47(m,5H),2.40–2.32(m,1H),2.04–1.96(m,1H),1.88–1.78(m,4H),1.68–1.53(m,4H),1.19(t,J=7.3Hz,3H). 13C NMR(151MHz,DMSO-d6)δ172.83,170.18,169.65,167.76,167.22,166.52,154.00 ,152.27,151.74,150.14,136.42,135.35,134.02,131.48,124.77,118.27,116.6 4,111.66,111.58,104.76,66.44,61.14,55.69,50.53,48.88,48.64,48.49,47. 57,44.43,44.21,41.94,32.18,30.99,28.19,24.20,22.24,10.95.HRMS(ESI)for C 43 H 54 O8N 11 [M+H] + , Calcd: 852.4151, Found: 852.4143. HPLC analysis: MeCN-H2O (0.1% TFA), RT=8.481min, 98.87% purity.

[0247] Example 23: Preparation of 3-((4-(4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)butyryl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-10b)

[0248]

[0249] The synthesis method is the same as Example 4 (yellow solid, 69 mg, 39%). 1H NMR(500MHz,DMSO-d6)δ11.06(s,1H),11.02(s,1H),7.57(d,J=8.3Hz,1H),7.53(d,J=3.3Hz,1H),7.26(dd,J=8.6,2.3Hz,1H),7.22(d,J=3.3Hz,1H),7.16(t,J=5.5Hz,1H),7.02(d,J=2.3Hz,1H),6.96(d,J=2.1Hz,1H),6.86(dd,J=8.4,2.1Hz,1H),6.81(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),5.03(dd,J=12.9,5.4Hz,1H),4.16–4.07(m,1H),3.94–3.89(m,2H),3.81(s,3H),3.49–3.40(m,4H),3.40–3.29(m,4H),3.19(q,J=6.6Hz,2H),2.92–2.81(m,1H),2.61–2.51(m,5H),2.49–2.39(m,7H),2.35–2.27(m,1H),2.01–1.95(m,1H),1.88–1.82(m,2H),1.82–1.75(m,4H),1.68–1.58(m,2H),1.58–1.50(m,2H),1.19(t,J=7.4Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ172.88,170.23,170.05,169.68,167.73,167.19,154.46,152.29,151.76,150.16,136.38,135.39,134.25,131.52,125.18,118.29,115.90,111.67,111.56,104.71,66.48,61.19,55.70,50.51,49.00,48.63,48.57,47.60,45.22,42.04,41.43,32.20,31.01,29.48,28.13,24.24,23.95,22.27,10.99.HRMS(ESI)for C 45 H 58 O8N 11 [M+H] + ,Calcd:880.4464,Found:880.4456.HPLC analysis:MeCN-H2O(0.1%TFA),RT=8.632min,97.81%purity.

[0250] Example 24: Preparation of 3-((4-(4-(4-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)hexanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-10c)

[0251]

[0252] The synthesis method is the same as Example 4 (yellow solid, 96 mg, 53%). 1 H NMR (500MHz, DMSO-d6) δ11.06(s,1H),11.02(s,1H),7.56(d,J=8.3Hz,1H),7.53(d,J=3 .3Hz,1H),7.26(dd,J=8.6,2.3Hz,1H),7.22(d,J=3.3Hz,1H),7.11(t,J=5.3Hz,1H),7. 02(d,J=2.3Hz,1H),6.94(d,J=2.1Hz,1H),6.84(dd,J=8.4,2.1Hz,1H),6.81(d,J=8.7H z,1H),6.78(d,J=7.5Hz,1H),5.02(dd,J=12.8,5.4Hz,1H),4.11(dtt,J=11.4,8.0,4.0H z,1H),3.94–3.89(m,2H),3.80(s,3H),3.46–3.39(m,4H),3.39–3.35(m,2H),3.32–3.2 8(m,2H),3.15(q,J=6.6Hz,2H),2.86(ddd,J=16.9,13.9,5.3Hz,1H),2.61–2.51(m,4H), 2.49–2.41(m,6H),2.33–2.26(m,3H),2.01–1.94(m,1H),1.88–1.81(m,2H),1.80–1.74 (m,2H),1.67–1.60(m,2H),1.60–1.50(m,6H),1.41–1.34(m,2H),1.19(t,J=7.4Hz,3H). 13C NMR (151MHz, DMSO-d6) δ172.82,170.35,170.18,169.65,167.72,167.16,154.48,152. 27,151.74,150.14,136.42,135.35,134.22,131.49,125.12,118.26,115.78,111.66, 111.58,104.77,66.44,61.10,55.69,50.50,49.12,48.62,48.60,47.57,45.39,42.40 ,41.40,32.19,30.99,28.19,28.09,26.32,24.59,24.20,22.24,10.95.HRMS(ESI)for C 47 H 62 O8N 11 [M+H] + , Calcd: 908.4777, Found: 908.4766. HPLC analysis: MeCN-H2O (0.1% TFA), RT=8.996min, 97.99% purity.

[0253] Example 25: Preparation of 3-((4-(4-(4-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)octanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-10d)

[0254]

[0255] The synthesis method is the same as Example 4 (yellow solid, 50 mg, 27%). 1HNMR(500MHz,DMSO-d6)δ11.06(s,1H),11.02(s,1H),7.56(dd,J=8.3,1.8Hz,1H),7.53(s,1H),7.26(d,J=8.5Hz,1H),7.22(s,1H),7.10(t,J=5.7Hz,1H),7.02(s,1H),6.94(s,1H),6.87–6.76(m,3H),5.02(dd,J=12.7,5.4Hz,1H),4.19–4.07(m,1H),3.95–3.88(m,2H),3.80(s,3H),3.47–3.39(m,4H),3.39–3.34(m,2H),3.34–3.29(m,2H),3.15(q,J=6.7Hz,2H),2.92–2.82(m,1H),2.62–2.51(m,4H),2.50–2.40(m,6H),2.37–2.30(m,1H),2.28(t,J=7.5Hz,2H),2.02–1.94(m,1H),1.88–1.82(m,2H),1.81–1.75(m,2H),1.68–1.59(m,2H),1.59–1.52(m,4H),1.52–1.45(m,2H),1.40–1.26(m,6H),1.19(t,J=7.4Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ172.82,170.44,170.19,169.65,167.72,167.16,154.48,152.27,151.74,150.14,136.42,135.34,134.22,131.49,125.11,118.26,115.77,111.66,111.58,104.77,66.44,61.11,55.69,50.51,49.12,48.61,47.57,45.39,42.47,41.38,32.22,32.18,30.99,28.75,28.61,28.20,28.18,26.45,24.80,24.20,22.24,10.96.HRMS(ESI)for C 49 H 66 O8N 11 [M+H] + ,Calcd:936.5090,Found:936.5071.HPLC analysis:MeCN-H2O(0.1%TFA),RT=9.555min,95.67%purity.

[0256] Example 26: Preparation of 3-((4-(4-(4-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)decanoyl)piperazin-1-yl))piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-10e)

[0257] Step 1: 10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)decanoic acid (2-38e)

[0258]

[0259] 10-Aminodecanoic acid 2-32f (5.0 g, 26.7 mmol) was dissolved in 50 mL of thionyl chloride, reacted at room temperature for 2 hours, concentrated under reduced pressure to obtain a crude product, and then 100 mL of anhydrous tert-butyl alcohol was added and stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the solvent was removed by rotary evaporation under reduced pressure to obtain a crude compound 2-34f. 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione 2-23 (1.1 g, 3.8 mmol), compound 2-34f (1.0 g, 4.2 mmol) and N,N-diisopropylethylamine (DIPEA, 2 mL, 11.4 mmol) were dissolved in 20 mL of N-methylpyrrolidone and heated to 120 ° C for 5 hours. After the reaction was completed by TLC monitoring, the heating was stopped and the system was cooled to room temperature. 100 mL of ice water was added to the system, and ethyl acetate was extracted three times. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum rotary evaporation. The residue was separated and purified by column chromatography (dichloromethane / methanol = 100: 1) to obtain a yellow solid compound 2-36e. The obtained compound 2-36e was dissolved in 10 mL of dichloromethane, 2 mL of trifluoroacetic acid was added, and the reaction was carried out at room temperature for 2 hours. After the reaction was completed by TLC monitoring, the solvent was removed by vacuum rotary evaporation, and the residue was separated and purified by column chromatography (dichloromethane / methanol = 25: 1) to obtain a yellow solid compound 2-38e (120 mg, 75%). 1H NMR (500MHz, DMSO-d6) δ11.98(s,1H),11.05(s,1H),7.55(d,J=8.4Hz,1H),7.14–7.05(m,1H),6.93(d,J=2.1 Hz,1H),6.84(dd,J=8.4,2.1Hz,1H),5.02(dd,J=12.7,5.4Hz,1H),3.17–3.12(m,2H),2.87(ddd,J=16.6,13. 7,5.4Hz,1H),2.61–2.54(m,1H),2.53–2.51(m,1H),2.18(t,J=7.3Hz,2H),2.03–1.95(m,1H),1.61–1.51(m, 2H),1.51–1.45(m,2H),1.39–1.33(m,2H),1.32–1.28(m,2H),1.27–1.23(m,6H).MS(ESI),m / z:466.2[M+Na] + .

[0260] Step 2: 3-((4-(4-(4-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)decanoyl)piperazin-1-yl))piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-10e)

[0261]

[0262] Compound 2-5 (108 mg, 0.2 mmol), compound 2-38e (87 mg, 0.2 mmol) and polypeptide condensation agent 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU, 91 mg, 0.24 mmol) were dissolved in 5 mL N,N-dimethylformamide, and N,N-diisopropylethylamine (DIPEA, 105 μL, 0.6 mmol) was added and reacted at room temperature for 6 hours. After the reaction was completed by TLC monitoring, 30 mL of ice water was added to the system, and ethyl acetate was extracted three times. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum rotary evaporation. The residue was separated and purified by column chromatography (dichloromethane / methanol = 15:1) to obtain yellow solid compound 2-10e (119 mg, 62%). 1H NMR(500MHz,DMSO-d6)δ11.06(s,1H),11.01(s,1H),7.55(d,J=8.3Hz,1H),7.53(d,J=3.2Hz,1H),7.26(dd,J=8.6,2.3Hz,1H),7.22(d,J=3.2Hz,1H),7.10(t,J=5.3Hz,1H),7.02(d,J=2.3Hz,1H),6.93(d,J=2.1Hz,1H),6.84(dd,J=8.6,2.1Hz,1H),6.81(d,J=8.7Hz,1H),6.78(d,J=7.6Hz,1H),5.02(dd,J=12.7,5.4Hz,1H),4.15–4.06(m,1H),3.94–3.89(m,2H),3.80(s,3H),3.44–3.36(m,6H),3.33–3.30(m,2H),3.14(q,J=6.6Hz,2H),2.87(ddd,J=16.7,13.6,5.2Hz,1H),2.61–2.52(m,5H),2.49–2.42(m,5H),2.37–2.29(m,1H),2.27(t,J=7.5Hz,2H),2.02–1.95(m,1H),1.87–1.82(m,2H),1.81–1.75(m,2H),1.67–1.60(m,2H),1.60–1.52(m,4H),1.49–1.44(m,2H),1.40–1.33(m,2H),1.32–1.29(m,2H),1.29–1.25(m,6H),1.18(t,J=7.3Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ172.89,170.47,170.25,169.69,167.76,167.20,154.50,152.28,151.77,150.17,136.41,135.39,134.24,131.50,125.15,118.26,115.78,111.67,111.56,104.72,66.49,61.15,55.70,50.53,49.13,48.64,45.39,42.53,41.38,32.26,32.21,31.02,28.99,28.89,28.81,28.27,28.19,26.58,24.88,24.25,22.28,10.98.HRMS(ESI)for C 51 H 70 O8N 11 [M+H]+ , Calcd: 964.5403, Found: 964.5395. HPLC analysis: MeCN-H2O (0.1% TFA), RT=10.152min, 98.56% purity.

[0263] Example 27: Preparation of 3-((4-(4-(4-(12-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)dodecanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-10f)

[0264]

[0265] The synthesis method is the same as Example 4 (yellow solid, 67 mg, 34%). 1 H NMR (500MHz, DMSO-d6) δ11.05(s,1H),11.02(s,1H),7.55(d,J=8.3Hz,1H),7.52(d,J=3.3 Hz,1H),7.26(dd,J=8.6,2.3Hz,1H),7.21(d,J=3.3Hz,1H),7.09(t,J=5.4Hz,1H),7.02(d ,J=2.3Hz,1H),6.93(d,J=2.1Hz,1H),6.86–6.81(m,1H),6.81(d,J=8.7Hz,1H),6.77(d,J =7.5Hz,1H),5.02(dd,J=12.7,5.4Hz,1H),4.17–4.05(m,1H),3.95–3.88(m,2H),3.80(s,3 H),3.45–3.38(m,4H),3.37–3.29(m,4H),3.14(q,J=6.5Hz,2H),2.92–2.81(m,1H),2.62– 2.51(m,4H),2.49–2.40(m,6H),2.35–2.29(m,1H),2.26(t,J=7.5Hz,2H),2.02–1.94(m,1 H),1.88–1.81(m,2H),1.81–1.75(m,2H),1.69–1.60(m,2H),1.60–1.50(m,4H),1.50–1.4 3(m,2H),1.40–1.32(m,2H),1.32–1.28(m,2H),1.28–1.22(m,10H),1.19(t,J=7.4Hz,3H). 13C NMR (126MHz, DMSO-d6) δ172.87,170.45,170.23,169.68,167.74,167.19,154.49,152.27,151. 75,150.16,136.40,135.37,134.23,131.49,125.14,118.26,115.77,111.67,111.55,104.71,6 6.47,61.14,55.69,50.53,49.12,48.63,47.60,45.40,42.50,41.39,32.25,32.20,31.01,29. 07,29.00,28.92,28.85,28.81,28.26,28.18,26.57,24.89,24.23,22.27,10.97.HRMS(ESI)for C 53 H 74 O8N 11 [M+H] + , Calcd: 992.5716, Found: 992.5725. HPLC analysis: MeCN-H2O (0.1% TFA), RT=10.940min, 96.53% purity.

[0266]

[0267] Reaction reagents and conditions: (a) TFA, DCM, room temperature, 2h; (b) HATU, DIPEA, DMF, room temperature, 6h, 23%; (c) TFA, DCM, room temperature, 2h; (d) HATU, DIPEA, DMF, room temperature, 6h, 39%; (e) iodomethane, potassium carbonate, DMF, 80℃, 4h; (f) TFA, DCM, room temperature, 2h, 88%; (g) HATU, DIPEA, DMF, room temperature, 6h, 33%.

[0268] Example 28: Preparation of 3-((4-(4-(4-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-4-yl)oxy)acetyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-7aa)

[0269]

[0270] The synthesis method is the same as Example 4 (yellow solid, 38 mg, 23%). 1H NMR(500MHz,DMSO-d6)δ11.02(s,1H),10.99(s,1H),7.53(d,J=3.2Hz,1H),7.46(t,J=7.8Hz,1H),7.32(d,J=7.4Hz,1H),7.27(dd,J=8.6,2.3Hz,1H),7.23(d,J=3.3Hz,1H),7.15(d,J=8.2Hz,1H),7.02(d,J=2.3Hz,1H),6.82(d,J=8.6Hz,1H),6.79(d,J=7.5Hz,1H),5.12(dd,J=13.3,5.1Hz,1H),5.02(s,2H),4.44–4.23(m,2H),4.16–4.07(m,1H),3.95–3.89(m,2H),3.81(s,3H),3.49–3.42(m,4H),3.40–3.33(m,4H),2.92(ddd,J=17.3,13.6,5.2Hz,1H),2.62–2.54(m,5H),2.54–2.41(m,5H),2.38–2.31(m,1H),2.04–1.97(m,1H),1.88–1.82(m,2H),1.82–1.77(m,2H),1.68–1.60(m,2H),1.59–1.52(m,2H),1.19(t,J=7.4Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ172.94,171.08,169.69,168.05,165.30,153.40,152.27,151.76,150.17,136.38,135.39,133.31,131.49,129.77,129.68,118.26,115.39,115.05,111.67,111.55,104.70,66.48,65.92,61.18,55.68,54.95,51.66,50.51,48.87,48.47,47.60,45.17,44.51,41.71,32.21,31.28,28.16,24.24,22.44,10.96.HRMS(ESI)for C 43 H 55 N 10 O8[M+H] + ,Calcd:839.4199,Found:839.4203.HPLC analysis:MeCN-H2O(0.1%TFA),RT=8.081min,96.69%purity.

[0271] Example 29: Preparation of 3-((4-(4-(4-(2-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindol-4-yl)oxy)acetyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-7ab)

[0272]

[0273] The synthesis method is the same as Example 4 (yellow solid, 65 mg, 39%). 1 H NMR(600MHz,DMSO-d6)δ11.02(s,1H),10.96(s,1H),7.53(d,J=3.1Hz,1H),7.5 2–7.49(m,1H),7.26(dd,J=8.6,2.3Hz,1H),7.22(d,J=3.2Hz,1H),7.12(d,J=7 .5Hz,1H),7.02(d,J=2.4Hz,1H),6.89(d,J=8.3Hz,1H),6.82(d,J=8.6Hz,1H), 6.78(d,J=7.5Hz,1H),5.06(dd,J=13.3,5.2Hz,1H),5.00(s,2H),4.40–4.21(m, 2H),4.16–4.08(m,1H),3.94–3.90(m,2H),3.81(s,3H),3.53–3.42(m,4H),3.3 9–3.33(m,4H),2.91(ddd,J=17.3,13.7,5.5Hz,1H),2.62–2.54(m,5H),2.52–2. 47(m,4H),2.41–2.31(m,2H),2.00–1.95(m,1H),1.88–1.82(m,2H),1.81–1.76 (m,2H),1.63(qd,J=12.4,4.5Hz,2H),1.59–1.52(m,2H),1.19(t,J=7.4Hz,3H). 13C NMR(151MHz,DMSO-d6)δ172.91,171.14,169.64,166.54,165.40,155.64,152.27,151.7 3,150.13,144.58,136.40,135.35,133.09,131.47,118.96,118.27,115.68,112.05,111 .66,111.56,104.74,66.43,66.39,61.14,55.68,54.91,51.30,50.50,48.91,48.44,47. 56,46.51,44.67,41.76,32.18,31.24,28.17,28.10,24.19,22.45,10.95.HRMS(ESI)for C 43 H 55 N 10 O8[M+H] + , Calcd: 839.4199, Found: 839.4186. HPLC analysis: MeCN-H2O (0.1% TFA), RT=8.198min, 97.28% purity.

[0274] Example 30: Preparation of 6-ethyl-3-((3-methoxy-4-(4-(4-(2-((2-(1-methyl-2,6-dioxopiperidin-3-yl)-1-oxoisoindole-4-yl)oxy)acetyl)piperazin-1-yl)piperidin-1-yl)phenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-7ac)

[0275] Step 1: 2-((2-(1-methyl-2,6-dioxopiperidin-3-yl)-1-oxoisoindole-4-yl)oxy)acetic acid (2-44)

[0276]

[0277] Tert-butyl 2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindole-4-yl)oxy)acetate 2-39 (374 mg, 1 mmol), iodomethane (62 μL, 1 mmol) and potassium carbonate (415 mg, 3 mmol) were dissolved in 15 mL N, N-dimethylformamide and reacted at 80°C for 4 hours. After the reaction was completed, 30 mL of ice water was added to the system, extracted three times with ethyl acetate, the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum rotary evaporation to obtain a white solid compound 2-43. Compound 2-43 was dissolved in 10 mL of dichloromethane, 2 mL of trifluoroacetic acid was added, and the reaction was carried out at room temperature for 2 hours. After the reaction was completed by TLC monitoring, the solvent was removed by vacuum rotary evaporation, and the residue was separated and purified by column chromatography (dichloromethane / methanol = 25:1) to obtain a white solid compound 2-44 (155 mg, 88%). 1 H NMR (500MHz, DMSO-d6) δ13.11(s,1H),7.47(t,J=7.8Hz,1H),7.34(d,J=7.5Hz,1H),7.17(d,J=8.1Hz,1H),5.18(dd,J=13.5,5.1Hz,1H) ,4.85(s,2H),4.44–4.20(m,2H),3.04–2.94(m,4H),2.78–2.71(m,1H),2.48–2.39(m,1H),2.06–1.97(m,1H).MS(ESI),m / z:333.0[M+H] + .

[0278] Step 2: 6-ethyl-3-((3-methoxy-4-(4-(4-(2-((2-(1-methyl-2,6-dioxopiperidin-3-yl)-1-oxoisoindole-4-yl)oxy)acetyl)piperazin-1-yl)piperidin-1-yl)phenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (2-7ac)

[0279]

[0280] The synthesis method is the same as Example 4 (yellow solid, 56 mg, 33%). 1H NMR(500MHz,DMSO-d6)δ11.02(s,1H),7.53(d,J=2.8Hz,1H),7.47(t,J=7.8Hz,1H),7.33(d,J=7.4Hz,1H),7.27(dd,J=8.6,2.3Hz,1H),7.22(d,J=3.2Hz,1H),7.16(d,J=8.2Hz,1H),7.02(d,J=2.3Hz,1H),6.82(d,J=8.7Hz,1H),6.78(d,J=7.5Hz,1H),5.18(dd,J=13.4,5.1Hz,1H),5.01(s,2H),4.43–4.24(m,2H),4.15–4.07(m,1H),3.95–3.89(m,2H),3.81(s,3H),3.50–3.42(m,4H),3.39–3.30(m,4H),3.01(s,3H),3.03–2.95(m,1H),2.62–2.54(m,4H),2.54–2.51(m,4H),2.47–2.40(m,1H),2.40–2.29(m,1H),2.05–1.95(m,2H),1.88–1.77(m,4H),1.67–1.54(m,4H),1.19(t,J=7.4Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ171.98,170.70,169.69,168.08,165.30,153.40,152.28,151.76,150.17,136.40,135.39,133.29,131.51,129.82,129.71,118.27,115.44,115.09,111.67,111.56,104.71,66.48,65.97,61.18,55.69,54.96,52.17,50.54,48.91,48.50,47.61,45.16,44.56,41.75,32.21,31.42,28.18,26.61,24.24,21.71,10.98.HRMS(ESI)for C 44 H 56 N 10 NaO8[M+Na] + ,Calcd:875.4175,Found:875.4174.HPLC analysis:MeCN-H2O(0.1%TFA),RT=8.383min,99.23%purity.

[0281] Example 31: (E)-3-((4-(4-(4-(3-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)acryloyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-99)

[0282]

[0283] The synthesis method is the same as Example 4, yellow solid. 1 H NMR (500MHz, DMSO-d6) δ11.14(s,1H),11.02(s,1H),8.44(s,1H),8.16(d,J=7.8Hz,1H),7.94(d,J=7.7Hz,1H),7.65(s,2H),7.53(d,J=3.3Hz ,1H),7.27(dd,J=8.5,2.3Hz,1H),7.22(d,J=3.2Hz,1H),7.02(d,J=2.3Hz,1H),6.83(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),5.18(dd,J=12. 8,5.4Hz,1H),4.15–4.08(m,1H),3.94–3.89(m,2H),3.81(s,3H),3.79 –3.74(m,2H),3.61–3.56(m,2H),3.39–3.33(m,5H),2.90(ddd,J=16.7 ,13.7,5.4Hz,1H),2.65–2.52(m,9H),2.40–2.33(m,1H),2.10–2.05(m ,1H),1.84(t,J=12.5Hz,4H),1.68–1.56(m,4H),1.19(t,J=7.3Hz,3H).

[0284] Example 32: 3-((4-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-carbonyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-100)

[0285]

[0286] The synthesis method is the same as Example 4, yellow solid. 1H NMR (500MHz, DMSO-d6) δ11.02(s,2H),7.79(d,J=7.7Hz,1H),7.64(s,1H),7.55–7.49(m,2H),7.26(dd,J=8.5,2.3Hz,1H),7.22(d,J =3.3Hz,1H),7.02(d,J=2.3Hz,1H),6.81(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),5.14(dd,J=13.3,5.1Hz,1H),4.54–4.34(m,2H),4 .17–4.07(m,1H),3.94–3.89(m,2H),3.80(s,3H),3.70–3.62(m,2H),3.44–3.28(m,6H),2.92(ddd,J=18.1,13.6,5.6Hz,1H),2.65– 2.55(m,6H),2.54–2.46(m,3H),2.46–2.31(m,2H),2.05–1.99(m,1H),1.87–1.77(m,4H),1.68–1.52(m,4H),1.18(t,J=7.3Hz,3H).

[0287] Example 33: 3-((4-(4-(4-(6-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-6-azaspiro[2.5]octane-1-carbonyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-150)

[0288]

[0289] The synthesis method is the same as Example 4, yellow solid. 1H NMR(500MHz,DMSO-d6)δ11.08(s,1H),11.01(s,1H),7.69(dd,J=8.4,7.1Hz,1H),7.52(d, J=3.3Hz,1H),7.35(dd,J=12.6,7.8Hz,2H),7.26(dd,J=8.6,2.3Hz,1H),7.21(d,J=3.3Hz ,1H),7.02(d,J=2.4Hz,1H),6.82(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),5.09(dd,J=12. 8,5.4Hz,1H),4.14–4.06(m,1H),3.95–3.88(m,2H),3.80(s,3H),3.72–3.60(m,2H),3.60– 3.52(m,1H),3.47–3.39(m,1H),3.39–3.29(m,7H),3.26–3.15(m,2H),2.87(ddd,J=16.9, 13.8,5.4Hz,1H),2.68–2.51(m,7H),2.48–2.39(m,2H),2.37–2.29(m,1H),2.06–1.99(m, 1H),1.98–1.93(m,1H),1.88–1.82(m,2H),1.82–1.76(m,2H),1.76–1.71(m,1H),1.70–1. 58(m,5H),1.58–1.50(m,2H),1.19(t,J=7.4Hz,3H),1.11–1.06(m,1H),0.80–0.76(m,1H).

[0290] Example 34: 3-((4-(4-(6-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-6-azaspiro[2.5]octane-1-carbonyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-153)

[0291]

[0292] The synthesis method is the same as Example 4, yellow solid. 1H NMR(500MHz,DMSO-d6)δ11.06(s,1H),11.01(s,1H),7.66(dd,J=8.6,1.2Hz,1H),7.52( d,J=3.3Hz,1H),7.35(t,J=2.5Hz,1H),7.28–7.23(m,2H),7.21(d,J=3.2Hz,1H),7.02( dd,J=2.4,1.2Hz,1H),6.78(dd,J=11.8,8.1Hz,2H),5.05(dd,J=12.9,5.4Hz,1H),4.10 (dtt,J=11.4,7.9,4.0Hz,1H),3.94–3.88(m,2H),3.79(s,3H),3.66–3.60(m,2H),3.59– 3.53(m,3H),3.50–3.45(m,1H),3.43–3.35(m,4H),3.32–3.26(m,3H),2.91–2.80(m,1H ),2.61–2.51(m,5H),2.49–2.42(m,3H),2.42–2.35(m,1H),2.29(t,J=11.5Hz,1H),2.00 –1.93(m,2H),1.87–1.81(m,2H),1.78–1.71(m,2H),1.62(qd,J=12.3,4.6Hz,4H),1.58 –1.44(m,4H),1.18(t,J=7.4Hz,3H),1.09(t,J=4.6Hz,1H),0.79(dd,J=7.9,3.9Hz,1H).

[0293] Example 35: 3-((4-(4-(4-(8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-8-oxooctanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-136)

[0294]

[0295] The synthesis method is the same as Example 4, yellow solid. 1H NMR(500MHz,DMSO-d6)δ11.02(s,2H),9.76(s,1H),7.81(dd,J=7.3,1.8Hz, 1H),7.55–7.51(m,1H),7.51–7.46(m,2H),7.26(dd,J=8.6,2.3Hz,1H),7.21 (d,J=3.3Hz,1H),7.02(d,J=2.3Hz,1H),6.82(d,J=8.6Hz,1H),6.78(d,J=7. 5Hz,1H),5.14(dd,J=13.3,5.1Hz,1H),4.43–4.30(m,2H),4.17–4.06(m,1H) ,3.95–3.89(m,2H),3.80(s,3H),3.46–3.39(m,4H),3.39–3.28(m,4H),2.9 7–2.86(m,1H),2.64–2.54(m,3H),2.49–2.41(m,5H),2.38–2.33(m,3H),2.3 3–2.26(m,4H),2.05–1.99(m,1H),1.88–1.82(m,2H),1.81–1.75(m,2H),1.6 8–1.57(m,4H),1.57–1.46(m,4H),1.35–1.30(m,4H),1.19(t,J=7.3Hz,3H).

[0296] Example 36: 3-((4-(4-(4-(6-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-6-oxohexanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-140)

[0297]

[0298] The synthesis method is the same as Example 4, yellow solid. 1H NMR (500MHz, DMSO-d6) δ11.01(d,J=3.4Hz,2H),9.79(s,1H),7.80(dd,J=7.2,1.7Hz,1H),7.55–7.45(m,3H),7.26(dd,J=8.6,2.3Hz,1H),7.21(d ,J=2.7Hz,1H),7.02(d,J=2.3Hz,1H),6.82(d,J=8.7Hz,1H),6.77(d,J=7 .5Hz,1H),5.15(dd,J=13.3,5.1Hz,1H),4.43–4.30(m,2H),4.15–4.06(m ,1H),3.96–3.88(m,2H),3.81(s,3H),3.48–3.40(m,3H),3.40–3.33(m, 4H),3.31–3.23(m,2H),2.98–2.87(m,1H),2.64–2.54(m,3H),2.49–2.42 (m,4H),2.42–2.27(m,7H),2.06–1.99(m,1H),1.88–1.82(m,2H),1.81–1 .73(m,2H),1.68–1.59(m,4H),1.59–1.50(m,4H),1.19(t,J=7.3Hz,3H).

[0299] Example 37: 3-((4-(4-(4-(8-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-4-yl)amino)-8-oxooctanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-142)

[0300]

[0301] The synthesis method is the same as Example 4, yellow solid. 1H NMR(500MHz,DMSO-d6)δ11.03(s,1H),11.01(s,1H),10.27(s,1H),8.32(d,J =8.2Hz,1H),7.57(t,J=7.9Hz,1H),7.52(d,J=3.3Hz,1H),7.28–7.23(m,2H) ,7.21(d,J=3.3Hz,1H),7.02(d,J=2.3Hz,1H),6.81(d,J=8.7Hz,1H),6.77(d ,J=7.5Hz,1H),5.07(dd,J=13.3,5.2Hz,1H),4.50–4.33(m,2H),4.16–4.07(m ,1H),3.95–3.89(m,2H),3.80(s,3H),3.45–3.33(m,9H),2.90(ddd,J=17.2, 13.6,5.4Hz,1H),2.65–2.55(m,3H),2.49–2.39(m,8H),2.34–2.25(m,3H),2. 06–2.00(m,1H),1.88–1.82(m,2H),1.81–1.75(m,2H),1.68–1.59(m,4H),1. 59–1.52(m,2H),1.52–1.45(m,2H),1.36–1.29(m,4H),1.19(t,J=7.3Hz,3H).

[0302] Example 38: 3-((4-(4-(4-(3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)ethoxy)propanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY7-47)

[0303]

[0304] The synthesis method is the same as Example 4, yellow solid. 1HNMR (500MHz, DMSO-d6) δ11.11(s,1H),11.02(s,1H),7.81(td,J=7.8,7.1,1. 5Hz,1H),7.54(d,J=8.7Hz,2H),7.46(dd,J=7.2,1.4Hz,1H),7.26(dt,J=8.6, 1.9Hz,1H),7.22(s,1H),7.02(t,J=1.9Hz,1H),6.81(d,J=8.8Hz,1H),6.78(d ,J=8.1Hz,1H),5.08(dd,J=12.6,5.3Hz,1H),4.34(t,J=4.5Hz,2H),4.16–4.0 7(m,1H),3.94–3.89(m,2H),3.82–3.79(m,5H),3.67–3.58(m,4H),3.54–3.49 (m,2H),3.48–3.29(m,8H),2.93–2.84(m,1H),2.65–2.51(m,7H),2.49–2.39( m,5H),2.34–2.24(m,1H),2.05–1.99(m,1H),1.88–1.82(m,2H),1.80–1.73(m ,2H),1.63(qd,J=12.3,4.4Hz,2H),1.58–1.49(m,2H),1.19(t,J=7.0Hz,3H).

[0305] Example 39: 3-((4-(4-(4-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-2-oxoethyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-17)

[0306]

[0307] The synthesis method is the same as Example 4, yellow solid. 1H NMR(400MHz,Chloroform-d)δ11.15(s,1H),10.74(s,1H),8.88(d,J=8.4Hz,1H),8.65(s,1H),7.77–7.69(m,1H),7.58(d,J=7.3Hz,1H),7.50(s, 1H),7.46(dd,J=8.6,2.3Hz,1H),6.94(d,J=2.3Hz,1H),6.87(d,J=8.6Hz,1H),5.27(s,1H),4.96(dd,J=12.0,5.7Hz,1H),4.63(d,J=7.3Hz,1H),4 .27–4.15(m,1H),4.06–3.98(m,2H),3.87(s,3H),3.58–3.48(m,4H),3.3 0–3.24(m,2H),3.14–2.96(m,4H),2.93–2.84(m,4H),2.81–2.73(m,3H), 2.63(t,J=11.5Hz,2H),2.52(q,J=7.4Hz,2H),2.24–2.15(m,2H),2.13–2 .02(m,4H),1.99–1.88(m,2H),1.61–1.51(m,2H),1.30(t,J=7.4Hz,3H).

[0308] Example 40: 3-((4-(4-(4-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-8-oxooctyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-18)

[0309]

[0310] The synthesis method is the same as Example 4, yellow solid. 1H NMR (400MHz, Chloroform-d) δ10.71(s,1H),9.48(s,1H),8.81(d,J=8.5Hz,1H),7.71(dd,J=8.5,7.3Hz,1H),7.55(d,J=7.3Hz,1H),7.50(s ,1H),7.45(dd,J=8.6,2.3Hz,1H),6.90(d,J=2.3Hz,1H),6.84(d,J=8.6Hz,1H),5.17(s,1H),4.98–4.89(m,1H),4.61(d,J=7.3Hz,1H),4.26 –4.15(m,1H),4.05–3.98(m,2H),3.86(s,3H),3.58–3.46(m,4H),2.9 3–2.85(m,1H),2.85–2.70(m,6H),2.61–2.43(m,8H),2.41–2.31(m,1H ),2.25–2.15(m,1H),2.12–2.05(m,2H),1.99–1.90(m,2H),1.88–1.70(m,8H),1.63–1.50(m,6H),1.45–1.35(m,4H),1.29(t,J=7.4Hz,3H).

[0311] Example 41: 3-((4-(4-(4-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-10-oxodecanyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-21)

[0312]

[0313] The synthesis method is the same as Example 4, yellow solid. 1H NMR(400MHz,Chloroform-d)δ10.72(s,1H),9.46(s,1H),8.83(d,J=8.4Hz,1H),7.75–7.67(m,1H),7.55(dd,J=7.3,0.8Hz,1H),7.5 2(s,1H),7.46(dd,J=8.6,2.3Hz,1H),6.90(d,J=2.3Hz,1H),6.83(d,J=8.6Hz,1H),5.15(s,1H),4.95(dd,J=12.3,5.4Hz,1H),4.61( d,J=7.4Hz,1H),4.26–4.15(m,1H),4.06–3.98(m,2H),3.86(s,3H),3.58–3.45(m,4H),3.01–2.64(m,10H),2.61–2.42(m,8H),2.25 –2.13(m,2H),2.12–2.04(m,2H),2.02–1.90(m,2H),1.89–1.65(m,8H),1.55–1.45(m,5H),1.45–1.33(m,6H),1.30(t,J=7.4Hz,3H).

[0314] Example 42: 3-((4-(4-(4-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-6-oxohexyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-22)

[0315]

[0316] The synthesis method is the same as Example 4, yellow solid. 1H NMR (400MHz, Chloroform-d) δ10.73(s,1H),9.45(s,1H),8.80(d,J=8.5Hz,1H),7.72(dd,J=8.6,7.4Hz,1H),7.56(d,J=7.2Hz,1H),7.51( s,1H),7.47(dd,J=8.6,2.3Hz,1H),6.89(d,J=2.3Hz,1H),6.83(d,J=8.7Hz,1H),5.17(s,1H),4.95(dd,J=12.3,5.5Hz,1H),4.61(d,J=7. 3Hz,1H),4.25–4.16(m,1H),4.06–3.98(m,2H),3.86(s,3H),3.57–3.47(m,4H),3.04–2.84(m,6H),2.83–2.69(m,4H),2.65–2.55(m,3H), 2.55–2.45(m,5H),2.23–2.17(m,1H),2.12–2.05(m,2H),2.03–1.94(m,2H),1.92–1.73(m,6H),1.50–1.35(m,6H),1.30(t,J=7.4Hz,3H).

[0317] Example 43: 3-((4-(4-(4-(12-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-12-oxododecyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-24)

[0318]

[0319] The synthesis method is the same as Example 4, yellow solid. 1H NMR (400MHz, Chloroform-d) δ10.73(s,1H),9.43(s,1H),8.83(d,J=8.5Hz,1H),7.71(dd,J=8.5,7.3Hz,1H),7.54(d,J=7.3Hz,1H),7.50(s,1H),7.46(d d,J=8.6,2.3Hz,1H),6.90(d,J=2.3Hz,1H),6.82(d,J=8.6Hz,1H),5.23(s, 1H),4.96(dd,J=12.3,5.3Hz,1H),4.63(d,J=7.3Hz,1H),4.26–4.13(m,1H), 4.02(dt,J=11.6,3.5Hz,2H),3.86(s,3H),3.57–3.46(m,4H),3.03–2.70(m ,10H),2.68–2.54(m,6H),2.51(q,J=7.4Hz,2H),2.45(t,J=7.6Hz,2H),2.26 –2.13(m,2H),2.12–2.04(m,3H),2.03–1.93(m,3H),1.91–1.81(m,3H),1.8 0–1.70(m,4H),1.69–1.49(m,6H),1.43–1.32(m,6H),1.29(t,J=7.3Hz,3H).

[0320] Example 44: 3-((4-(4-(4-(6-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-6-oxohexyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-26)

[0321]

[0322] The synthesis method is the same as Example 4, yellow solid. 1H NMR (400MHz, Chloroform-d) δ10.72(s,1H),8.61(s,1H),7.85(d,J=8.0Hz,1H),7.68(d,J=7.5Hz,1H),7.50(s,1H),7.49–7.42(m,2H),6.90(d,J= 2.3Hz,1H),6.83(d,J=8.6Hz,1H),5.33(s,1H),5.20(s,1H),5.14(dd,J= 13.2,5.1Hz,1H),4.62(d,J=7.3Hz,1H),4.50(q,J=16.7Hz,2H),4.25–4. 14(m,1H),4.05–3.98(m,2H),3.86(s,3H),3.57–3.45(m,4H),3.03–2.81 (m,8H),2.81–2.73(m,1H),2.72–2.62(m,2H),2.61–2.46(m,7H),2.43–2 .30(m,1H),2.25–2.15(m,2H),2.13–2.04(m,2H),2.04–1.97(m,2H),1.9 6–1.85(m,6H),1.62–1.53(m,2H),1.52–1.41(m,2H),1.30–1.27(m,3H).

[0323] Example 45: 3-((4-(4-(4-(8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-8-oxooctyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-27)

[0324]

[0325] The synthesis method is the same as Example 4, yellow solid. 1H NMR(400MHz,Chloroform-d)δ10.72(s,1H),8.36(s,1H),7.76–7.67(m,2H),7.52(s,1H),7.49–7.43(m,2H),6.90(d,J=2.3Hz,1H),6.83(d,J=8.6H z,1H),5.34(s,1H),5.24–5.12(m,2H),4.62(d,J=7.3Hz,1H),4.57–4.36 (m,2H),4.26–4.13(m,1H),4.05–3.98(m,2H),3.86(s,3H),3.57–3.44(m, 4H),2.94(s,6H),2.90–2.72(m,4H),2.72–2.61(m,2H),2.61–2.55(m,2H ),2.51(q,J=7.4Hz,3H),2.48–2.43(m,2H),2.41–2.29(m,2H),2.25–2.17 (m,2H),2.08(d,J=12.8Hz,3H),2.04–1.98(m,2H),1.98–1.90(m,3H),1. 87–1.81(m,2H),1.59–1.51(m,2H),1.42–1.35(m,4H),1.30–1.26(m,3H).

[0326] Example 46: 3-((4-(4-(4-(10-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-10-oxodecanyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-28)

[0327]

[0328] The synthesis method is the same as Example 4, yellow solid. 1H NMR (400MHz, CDCl3) δ10.71(s,1H),8.03(s,1H),7.72(d,J=7.4Hz,1H),7.60(d,J=7.9Hz,1H),7.53–7.49(m,1H),7.48–7.41(m,2H ),6.90(d,J=2.3Hz,1H),6.83(d,J=8.6Hz,1H),5.26–5.22(m,1H),5.19(dd,J=13.3,5.3Hz,1H),4.63(d,J=7.3Hz,1H),4.52–4.32( m,2H),4.25–4.14(m,1H),4.06–3.97(m,2H),3.85(s,3H),3.58–3.45(m,4H),2.91–2.64(m,10H),2.60–2.40(m,10H),2.30(qd,J=1 3.1,5.3Hz,2H),2.24–2.14(m,2H),2.11–2.03(m,2H),1.95–1.88(m,2H),1.85–1.64(m,6H),1.63–1.49(m,4H),1.32–1.26(m,6H).

[0329] Example 47: 3-((4-(4-(4-(12-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-12-oxododecyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-29)

[0330]

[0331] The synthesis method is the same as Example 4, yellow solid. 1H NMR(400MHz,Chloroform-d)δ10.71(s,1H),7.73(d,J=7.5Hz,1H),7.60(d,J=7.8Hz,1H),7.57–7.48(m,2H),7.48–7.43(m,2H),6.9 0(d,J=2.3Hz,1H),6.84(d,J=8.6Hz,1H),5.47–5.37(m,1H),5.35(t,J=4.9Hz,1H),5.22(dd,J=13.3,5.1Hz,1H),5.17(s,1H),4.61 (d,J=7.3Hz,1H),4.51–4.37(m,2H),4.27–4.15(m,1H),4.06–3.98(m,2H),3.86(s,3H),3.58–3.45(m,4H),2.95–2.62(m,10H),2.6 0–2.38(m,10H),2.39–2.31(m,1H),2.26–2.18(m,2H),2.12–1.98(m,4H),1.97–1.86(m,4H),1.86–1.69(m,10H),1.30–1.25(m,8H).

[0332] Example 48: 3-((4-(4-(4-(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)-5-oxopentyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-48)

[0333]

[0334] The synthesis method is the same as Example 4, yellow solid. 1H NMR (500MHz, DMSO-d6) δ11.11(s,1H),11.01(s,1H),10.55(s,1H),8.26(d,J=1.8 Hz,1H),7.91(dd,J=8.2,1.8Hz,1H),7.87(d,J=8.2Hz,1H),7.52(d,J=3.3Hz,1H), 7.25(dd,J=8.6,2.3Hz,1H),7.21(d,J=3.3Hz,1H),7.02(d,J=2.3Hz,1H),6.81(d ,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),5.12(dd,J=12.8,5.5Hz,1H),4.16–4.06(m, 1H),3.95–3.88(m,2H),3.80(s,3H),3.39–3.28(m,5H),2.88(ddd,J=16.9,13.8, 5.4Hz,1H),2.62–2.45(m,9H),2.41(t,J=7.3Hz,3H),2.39–2.32(m,2H),2.31–2.2 1(m,4H),2.04(dtd,J=12.9,5.3,2.2Hz,1H),1.88–1.82(m,2H),1.82–1.77(m,2H ),1.67–1.58(m,4H),1.57–1.50(m,2H),1.50–1.44(m,2H),1.19(t,J=7.4Hz,3H).

[0335] Example 49: 3-((4-(4-(4-(2-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-4-yl)amino)-2-oxoethyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-57)

[0336]

[0337] The synthesis method is the same as Example 4, yellow solid. 1H NMR (500MHz, DMSO-d6) δ11.34(s,1H),11.02(d,J=2.1Hz,2H),8.46(d,J=8.3Hz, 1H),7.57(t,J=7.9Hz,1H),7.53(d,J=3.3Hz,1H),7.26(dd,J=7.7,3.2Hz,2H),7. 22(d,J=3.3Hz,1H),7.03(d,J=2.3Hz,1H),6.83(d,J=8.6Hz,1H),6.78(d,J=7.5 Hz,1H),5.08(dd,J=13.2,5.2Hz,1H),4.46(d,J=17.5Hz,1H),4.32(d,J=17.5Hz, 1H),4.17–4.07(m,1H),3.95–3.90(m,2H),3.81(s,3H),3.40–3.29(m,4H),3.17 (s,2H),2.94(ddd,J=17.9,13.5,5.4Hz,1H),2.74–2.61(m,5H),2.61–2.51(m,8H ),2.42(qd,J=13.1,4.5Hz,1H),2.33–2.24(m,1H),2.09–2.02(m,1H),1.89–1.8 2(m,4H),1.63(qd,J=12.2,4.4Hz,2H),1.59–1.51(m,2H),1.19(t,J=7.4Hz,3H).

[0338] Example 50: 3-((4-(4-(4-(5-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-4-yl)amino)-5-oxopentyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-58)

[0339]

[0340] The synthesis method is the same as Example 4, yellow solid. 1H NMR (500MHz, DMSO-d6) δ11.03(s,1H),11.01(s,1H),10.26(s,1H),8.32(d,J=8. 3Hz,1H),7.57(t,J=7.9Hz,1H),7.52(d,J=3.3Hz,1H),7.28–7.22(m,2H),7.21( d,J=3.3Hz,1H),7.02(d,J=2.3Hz,1H),6.81(d,J=8.7Hz,1H),6.77(d,J=7.5Hz, 1H),5.07(dd,J=13.3,5.2Hz,1H),4.48(d,J=17.6Hz,1H),4.36(d,J=17.6Hz,1H ),4.17–4.06(m,1H),3.95–3.88(m,2H),3.80(s,3H),3.40–3.27(m,5H),2.90(d dd,J=17.3,13.6,5.4Hz,1H),2.65–2.54(m,4H),2.50–2.38(m,8H),2.40–2.31( m,2H),2.32–2.19(m,4H),2.03(dtd,J=12.5,5.2,2.2Hz,1H),1.88–1.81(m,2H) ,1.81–1.75(m,2H),1.69–1.57(m,4H),1.56–1.43(m,4H),1.19(t,J=7.4Hz,3H).

[0341] Example 51: 3-((4-(4-(4-(8-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-4-yl)amino)-8-oxooctyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LWY6-64)

[0342]

[0343] The synthesis method is the same as Example 4, yellow solid. 1H NMR(500MHz,DMSO-d6)δ11.04(s,1H),11.01(s,1H),10.27(s,1H),8.32(d,J=8 .3Hz,1H),7.57(t,J=7.9Hz,1H),7.52(d,J=3.1Hz,1H),7.28–7.23(m,2H),7.2 1(d,J=3.3Hz,1H),7.02(d,J=2.3Hz,1H),6.81(d,J=8.7Hz,1H),6.77(d,J=7.5 Hz,1H),5.07(dd,J=13.3,5.1Hz,1H),4.51–4.31(m,2H),4.16–4.06(m,1H),3.9 5–3.89(m,2H),3.80(s,3H),3.40–3.26(m,6H),2.94–2.85(m,1H),2.65–2.55( m,3H),2.49–2.43(m,4H),2.43–2.28(m,6H),2.27–2.16(m,4H),2.03(dtd,J=1 2.2,5.2,2.1Hz,1H),1.88–1.82(m,2H),1.81–1.74(m,2H),1.69–1.57(m,4H), 1.57–1.47(m,2H),1.43–1.35(m,2H),1.34–1.24(m,6H),1.19(t,J=7.4Hz,3H).

[0344] Example 52: 3-((4-(4-(4-(2-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)oxy)acetyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (JJ4-98)

[0345]

[0346] The synthesis method is the same as Example 4, yellow solid. 1H NMR (500MHz, DMSO-d6) δ11.03(s,1H),10.99(s,1H),7.54(s,1H),7.50(d,J=8.2Hz,1H),7.27(d,J=9.1Hz,1H),7.25–7.18(m,3H),7. 02(s,1H),6.83(d,J=8.7Hz,1H),6.80(d,J=7.4Hz,1H),5.11(dd,J=13.3,5.2Hz,1H),4.96(s,2H),4.41–4.21(m,2H),4.15–4.07(m, 1H),3.96–3.89(m,2H),3.81(s,3H),3.50(s,4H),3.44–3.35(m,4H),3.16(d,J=4.5Hz,2H),2.91(td,J=17.8,15.6,5.4Hz,1H),2.72 –2.63(m,2H),2.62–2.53(m,6H),2.44–2.34(m,1H),2.04–1.95(m,1H),1.90–1.80(m,4H),1.69–1.55(m,4H),1.19(t,J=7.3Hz,3H).

[0347] Example 53: 3-((4-(4-(4-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)acetyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (JJ4-99)

[0348]

[0349] The synthesis method is the same as Example 4, yellow solid. 1H NMR (500MHz, DMSO-d6) δ11.02(s,1H),10.98(s,1H),7.63(d,J=8.4Hz,1H),7.53(s,1H),7.27(dd,J=8.6,2.3Hz,1H),7.23(s,1H),7.14(d, J=2.2Hz,1H),7.05(dd,J=8.3,2.3Hz,1H),7.02(d,J=2.3Hz,1H),6.82(d,J=8.7Hz,1H),6.80(d,J=7.5Hz,1H),5.08(dd,J=13.3,5.1Hz,1H) ,4.95(s,2H),4.43–4.23(m,2H),4.16–4.08(m,1H),3.95–3.89(m,2H ),3.81(s,3H),3.50–3.43(m,4H),3.39–3.30(m,4H),2.95–2.86(m,1H ),2.62–2.55(m,5H),2.54–2.48(m,3H),2.41–2.32(m,2H),2.03–1.94(m,2H),1.88–1.77(m,4H),1.66–1.53(m,4H),1.19(t,J=7.4Hz,3H).

[0350] Example 54: 3-((4-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-5-yl)glycylamino)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (JJ5-13)

[0351]

[0352] The synthesis method is the same as Example 4, yellow solid. 1H NMR (500MHz, DMSO-d6) δ11.03(s,1H),10.96(s,1H),7.53(d,J=3.2Hz,1H),7.27(dd,J=8.6,3.5Hz,2H),7.23(d,J=3.3Hz,1H),7.01(dd,J=8.3,2 .2Hz,2H),6.91(d,J=2.2Hz,1H),6.82(d,J=8.7Hz,1H),6.79(d,J=7.5Hz,1H),5.91(t,J=5.1Hz,1H),5.08(dd,J=13.3,5.1Hz,1H),4.31–4.13(m, 2H),4.15–4.07(m,1H),3.98(d,J=5.0Hz,2H),3.95–3.90(m,2H),3.81(s ,3H),3.56–3.46(m,4H),3.40–3.31(m,4H),2.90(ddd,J=17.9,13.6,5.4 Hz,1H),2.62–2.54(m,5H),2.54–2.47(m,3H),2.43–2.31(m,2H),2.03–1 .93(m,2H),1.88–1.77(m,4H),1.68–1.52(m,4H),1.19(t,J=7.4Hz,3H).

[0353] Example 55: 3-((4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindol-5-yl)oxy)butanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (JJ5-14)

[0354]

[0355] The synthesis method is the same as Example 4, yellow solid. 1H NMR (500MHz, DMSO-d6) δ11.03(s,1H),11.00(s,1H),7.54(d,J=3.3Hz,1H),7.50(d,J=8.1Hz,1H),7.29–7.23(m,2H),7.23–7.18(m,2H),7.02(d,J =2.3Hz,1H),6.80(t,J=7.2Hz,2H),5.11(dd,J=13.3,5.1Hz,1H),4.40–4 .21(m,2H),4.16–4.09(m,1H),4.07(t,J=6.5Hz,2H),3.95–3.88(m,2H),3 .80(s,3H),3.49–3.40(m,4H),3.39–3.29(m,4H),2.90(ddd,J=18.3,13. 5,5.4Hz,1H),2.62–2.54(m,4H),2.48–2.43(m,6H),2.42–2.35(m,1H),2 .34–2.27(m,1H),2.03–1.92(m,4H),1.88–1.81(m,2H),1.79–1.72(m,2H ),1.63(qd,J=12.2,4.4Hz,2H),1.57–1.48(m,2H),1.19(t,J=7.4Hz,3H).

[0356] Example 56: 3-((4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-5-yl)oxy)butanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (JJ5-15)

[0357]

[0358] The synthesis method is the same as Example 4, yellow solid. 1H NMR(500MHz,DMSO-d6)δ11.04(s,1H),10.98(s,1H),7.62(d,J=8.4Hz,1H),7.54( d,J=3.3Hz,1H),7.27(dd,J=8.5,2.3Hz,1H),7.24(d,J=3.2Hz,1H),7.16(d,J=2.1 Hz,1H),7.05(dd,J=8.4,2.2Hz,1H),7.02(d,J=2.3Hz,1H),6.80(dd,J=8.3,2.9H z,2H),5.07(dd,J=13.3,5.1Hz,1H),4.41–4.23(m,2H),4.16–4.10(m,1H),4.08(t ,J=6.4Hz,2H),3.94–3.89(m,2H),3.80(s,3H),3.48–3.40(m,4H),3.39–3.28(m, 4H),2.90(ddd,J=18.1,13.6,5.4Hz,1H),2.61–2.53(m,3H),2.49–2.42(m,7H),2. 41–2.32(m,1H),2.32–2.24(m,1H),2.03–1.93(m,4H),1.88–1.81(m,2H),1.78–1. 72(m,2H),1.63(qd,J=12.2,4.4Hz,2H),1.57–1.48(m,2H),1.18(t,J=7.3Hz,3H).

[0359] Example 57: 3-((4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindol-4-yl)oxy)butanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (JJ5-16)

[0360]

[0361] The synthesis method is the same as Example 4, yellow solid. 1H NMR(500MHz,DMSO-d6)δ11.02(s,1H),10.96(s,1H),7.56–7.50(m,2H),7.25 (dd,J=8.6,2.3Hz,1H),7.22(d,J=3.3Hz,1H),7.10(d,J=7.5Hz,1H),7.05(d, J=8.3Hz,1H),7.02(d,J=2.3Hz,1H),6.81(d,J=8.6Hz,1H),6.78(d,J=7.5Hz ,1H),5.01(dd,J=13.3,5.1Hz,1H),4.40–4.21(m,2H),4.16–4.07(m,3H),3.9 6–3.89(m,2H),3.81(s,3H),3.50–3.41(m,4H),3.40–3.28(m,4H),2.90(ddd ,J=18.0,13.7,5.4Hz,1H),2.63–2.52(m,5H),2.49–2.41(m,5H),2.40–2.32( m,1H),2.32–2.24(m,1H),2.02–1.93(m,4H),1.88–1.82(m,2H),1.77–1.70(m ,2H),1.63(qd,J=12.1,4.5Hz,2H),1.56–1.46(m,2H),1.19(t,J=7.4Hz,3H).

[0362] Example 58: 3-((4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-4-yl)oxy)butanoyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (JJ5-17)

[0363]

[0364] The synthesis method is the same as Example 4, yellow solid. 1H NMR (500MHz, DMSO-d6) δ11.02(s,1H),10.98(s,1H),7.53(d,J=3.3Hz,1H),7.48(t,J=7.8Hz,1H),7.31(d,J=7.5Hz,1H),7.28–7.20(m,3H),7.0 2(d,J=2.3Hz,1H),6.81(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),5.12(dd,J=13.3,5.1Hz,1H),4.42–4.21(m,2H),4.17–4.07(m,3H),3.95–3.89 (m,2H),3.81(s,3H),3.48–3.40(m,4H),3.39–3.28(m,4H),2.92(ddd,J =18.7,13.6,5.4Hz,1H),2.64–2.54(m,3H),2.49–2.40(m,8H),2.29(t, J=11.6Hz,1H),2.03–1.94(m,4H),1.88–1.82(m,2H),1.78–1.73(m,2H) ,1.63(qd,J=12.1,4.4Hz,2H),1.58–1.48(m,2H),1.19(t,J=7.3Hz,3H).

[0365] Example 59: 3-((4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-4-yl)amino)butyryl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (JJ5-18)

[0366]

[0367] The synthesis method is the same as Example 4, yellow solid. 1H NMR(500MHz,DMSO-d6)δ11.02(s,1H),10.97(s,1H),7.53(d,J=3.3Hz,1H),7.33(t ,J=7.8Hz,1H),7.26(dd,J=8.6,2.3Hz,1H),7.22(d,J=3.3Hz,1H),7.02(d,J=2.3H z,1H),6.81(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),6.68–6.61(m,3H),4.99(dd,J =13.3,5.1Hz,1H),4.35–4.17(m,2H),4.16–4.07(m,1H),3.96–3.89(m,2H),3.81( s,3H),3.48–3.43(m,2H),3.43–3.37(m,3H),3.37–3.29(m,3H),3.23(q,J=6.7Hz, 2H),2.89(ddd,J=17.1,13.6,5.4Hz,1H),2.62–2.55(m,3H),2.49–2.42(m,6H),2. 41–2.34(m,3H),2.33–2.26(m,1H),2.01–1.94(m,1H),1.88–1.82(m,2H),1.81–1. 73(m,4H),1.63(qd,J=12.2,4.5Hz,2H),1.58–1.48(m,2H),1.19(t,J=7.4Hz,3H).

[0368] Example 60: 3-((4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)amino)butyryl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (JJ5-19)

[0369]

[0370] The synthesis method is the same as Example 4, yellow solid. 1H NMR(500MHz,DMSO-d6)δ11.02(s,1H),10.92(s,1H),7.53(d,J=3.3Hz,1H),7.39(d,J =8.3Hz,1H),7.26(dd,J=8.6,2.3Hz,1H),7.22(d,J=3.3Hz,1H),7.02(d,J=2.3Hz,1H ),6.81(d,J=8.6Hz,1H),6.78(d,J=7.5Hz,1H),6.66(dd,J=8.4,2.0Hz,1H),6.63(d, J=1.9Hz,1H),6.41(t,J=5.5Hz,1H),5.01(dd,J=13.3,5.1Hz,1H),4.27(d,J=16.6Hz, 1H),4.17–4.07(m,2H),3.96–3.89(m,2H),3.80(s,3H),3.49–3.40(m,4H),3.39–3.2 9(m,4H),3.10(q,J=6.6Hz,2H),2.88(ddd,J=17.1,13.6,5.4Hz,1H),2.63–2.52(m,4 H),2.49–2.39(m,7H),2.38–2.26(m,2H),1.97–1.90(m,1H),1.89–1.82(m,2H),1.82 –1.73(m,4H),1.63(qd,J=12.4,4.6Hz,2H),1.58–1.49(m,2H),1.19(t,J=7.3Hz,3H).

[0371] Example 61: 3-((4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyryl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (JJ5-20)

[0372]

[0373] The synthesis method is the same as Example 4, yellow solid. 1H NMR (500MHz, DMSO-d6) δ11.05–10.99(m,2H),7.53(d,J=3.3Hz,1H),7.31–7.25(m,2H),7.22(d,J=3.3Hz,1H),7.02(d,J=2.3Hz,1H),6.93(d,J=7. 4Hz,1H),6.84–6.75(m,3H),5.66(t,J=5.6Hz,1H),5.12(dd,J=13.3,5.1 Hz,1H),4.23(d,J=17.1Hz,1H),4.17–4.06(m,2H),3.95–3.89(m,2H),3. 81(s,3H),3.50–3.40(m,4H),3.39–3.29(m,4H),3.15(q,J=6.6Hz,2H),2 .92(ddd,J=17.2,13.6,5.4Hz,1H),2.66–2.54(m,4H),2.49–2.39(m,7H) ,2.31(ddd,J=13.3,9.0,5.2Hz,2H),2.06–2.00(m,1H),1.88–1.74(m,6H ),1.63(qd,J=12.3,4.6Hz,2H),1.58–1.49(m,2H),1.19(t,J=7.3Hz,3H).

[0374] Example 62: 3-((4-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)amino)butyryl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (JJ5-22)

[0375]

[0376] The synthesis method is the same as Example 4, yellow solid. 1H NMR(500MHz,DMSO-d6)δ11.02(s,1H),10.95(s,1H),7.53(d,J=3.3Hz,1H), 7.26(dd,J=8.3,3.1Hz,2H),7.22(d,J=3.3Hz,1H),7.02(d,J=2.3Hz,1H),6. 86(dd,J=8.3,2.3Hz,1H),6.83–6.76(m,3H),5.94(t,J=5.6Hz,1H),5.06(dd ,J=13.3,5.1Hz,1H),4.26(d,J=16.3Hz,1H),4.17–4.07(m,2H),3.95–3.88( m,2H),3.81(s,3H),3.48–3.41(m,4H),3.39–3.29(m,4H),3.07(q,J=6.6Hz ,2H),2.89(ddd,J=17.2,13.6,5.5Hz,1H),2.62–2.54(m,3H),2.49–2.38(m, 8H),2.37–2.27(m,2H),2.00–1.92(m,1H),1.88–1.82(m,2H),1.82–1.73(m, 4H), 1.63 (qd, J=12.4, 4.5Hz, 2H), 1.58–1.49 (m, 2H), 1.19 (t, J=7.4Hz, 3H).

[0377] Example 63: 3-((4-(4-(-4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)piperidin-4-yl)methyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (ZLC4-41)

[0378]

[0379] Step 1: tert-Butyl 4-((4-(1-(4-(3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)methyl)piperidine-1-carboxylate

[0380] To the DMF (3mL) system of intermediate 2-5 (65mg, 0.12mmol), 4-(2-bromomethyl)-1-Boc-piperazine (67mg, 0.24mmol) and potassium carbonate (33mg, 0.24mmol) were added in sequence, and the temperature was raised to 80°C. After the reaction was completed by TLC monitoring, it was concentrated under reduced pressure, extracted three times with dichloromethane / methanol system, the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by normal phase column chromatography to obtain a yellow solid (38mg, 43%). MS (ESI), m / z: 736.5 [M+H] + .

[0381] Step 2: 3-((4-(4-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)piperidin-4-yl)methyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (ZLC4-41)

[0382] Trifluoroacetic acid (1 mL) was added dropwise to the anhydrous dichloromethane system (2 mL) of the product of step 1 (68 mg, 0.093 mmol), stirred at room temperature overnight, concentrated under reduced pressure, and DMF (2 mL), DIPEA (0.1 mL), 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione (25.5 mg, 0.0925 mmol) were added in sequence, and the temperature was raised to 110° C. After the reaction was completed by TLC monitoring, it was concentrated under reduced pressure, extracted three times with a dichloromethane / methanol system, the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by normal phase column chromatography to obtain a yellow solid 4-41 (10 mg, 13%). 1H NMR (600MHz, DMSO-d6) δ11.08(s,1H),11.01(s,1H),7.67(dd,J=8.4,7.2Hz,1H),7.5 2(d,J=2.7Hz,1H),7.32(dd,J=11.0,7.8Hz,2H),7.26(dd,J=8.6,2.2Hz,1H),7.21(d ,J=2.8Hz,1H),7.02(d,J=2.3Hz,1H),6.82(d,J=8.7Hz,1H),6.78(d,J=7.5Hz,1H),5 .08(dd,J=12.8,5.5Hz,1H),4.11(d,J=7.4Hz,1H),3.92(d,J=11.2Hz,2H),3.81(s,3H ),3.69(d,J=11.6Hz,2H),3.51(s,5H),3.37(d,J=11.9Hz,5H),3.30(s,1H),2.87(t, J=10.3Hz,3H),2.61(d,J=3.8Hz,1H),2.58(q,J=7.4Hz,4H),2.54–2.51(m,5H),2.18( s,1H),2.05–1.96(m,2H),1.87–1.77(m,6H),1.71(s,1H),1.63(dd,J=11.7,3.6Hz,3 H),1.55(d,J=9.9Hz,1H),1.46(d,J=6.8Hz,1H),1.19(t,J=7.4Hz,3H).HRMS(ESI)for C 47 H 62 N 11 O7[M+H] + ,Calcd:892.4828,Found:892.4839.

[0383] Example 64: 3-((4-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)azetidin-3-yl)methyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (ZLC4-42)

[0384]

[0385] Step 1: tert-butyl 3-((4-(1-(4-(3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)methyl)azetidine-1-carboxylate (ZLC4-40)

[0386] 1-Boc-3-(bromomethyl)azetidine (125 mg, 0.5 mmol) and potassium carbonate (69 mg, 0.5 mmol) were added to the DMF (3 mL) system of intermediate 2-5 (134 mg, 0.25 mmol) in sequence, and the temperature was raised to 80°C. After the reaction was completed by TLC monitoring, it was concentrated under reduced pressure, extracted three times with dichloromethane / methanol system, the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by normal phase column chromatography to obtain a yellow solid (56 mg, 32%). MS (ESI), m / z: 707.5 [M+H] + .

[0387] Step 2: 3-((4-(4-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)azetidin-3-yl)methyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (ZLC4-42)

[0388] Trifluoroacetic acid (1 mL) was added dropwise to the anhydrous dichloromethane system (2 mL) of the product of step 1 (56 mg, 0.079 mmol), stirred at room temperature overnight, concentrated under reduced pressure, and then DMF (2 mL), DIPEA (0.1 mL), 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione (22 mg, 0.079 mmol) were added in sequence, and the temperature was raised to 110° C. After the reaction was completed by TLC monitoring, it was concentrated under reduced pressure, extracted three times with a dichloromethane / methanol system, the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by normal phase column chromatography to obtain a yellow solid 4-42 (8 mg, 12%). 1H NMR (600MHz, DMSO-d6) δ11.07–11.05(m,1H),11.02–11.00(m,1H),7.57–7.53(m,1H),7.53–7.51(m,1H),7.27–7.24(m,1H),7.22–7.20( m,1H),7.11–7.08(m,1H),7.03–7.01(m,1H),6.83–6.81(m,1H),6.79–6.76(m,2H),5.06–5.02(m,1H),4.32–4.25(m,2H),4.17–4.07(m,1 H),3.94–3.89(m,2H),3.86–3.79(m,5H),3.41–3.34(m,6H),2.93–2.82(m,2H),2.61–2.51(m,9H),2.49–2.46(m,1H),2.45–2.35(m,3H), 2.27(s,1H),2.02–1.95(m,2H),1.87–1.78(m,4H),1.63(dd,J=11.6,3.8Hz,2H),1.55(d,J=9.1Hz,2H),1.20–1.17(m,3H).HRMS(ESI)for C 45 H 58 N 11 O7[M+H] + ,Calcd:864.4515,Found:864.4523.

[0389] Example 65: 3-((4-(4-(4-(1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)azetidine-3-carbonyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LZ002-013)

[0390]

[0391] Step 1: 1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)azetidine-3-carboxylic acid 3-(4-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (3.25 g, 10 mmol), 3-azetidinecarboxylic acid (1.52 g, 15 mmol), palladium acetate (0.22 g, 1.0 mmol), RuPhos (0.93 g, 2.0 mmol) and NaO were prepared by heating 1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)azetidine-3-carboxylic acid to a molten salt. tBu (2.88 g, 30 mmol) was added to 1,4-dioxane (50 mL) system, replaced with nitrogen three times, and stirred. Stirred until uniform, heated to 85-95°C for 17 hours, concentrated under reduced pressure at 45°C, and purified by column chromatography (dichloromethane / methanol) to obtain a brown solid (0.72 g, 21%).

[0392] Step 2: 3-((4-(4-(4-(1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)azetidine-3-carbonyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LZ002-013)

[0393] The intermediate prepared in step 1 (0.52 g, 1.5 mmol), HATU (0.68 g, 1.8 mmol), DIPEA (0.78 g, 6.0 mmol), DMF (7.5 mL) and 2-5 (0.97 g, 1.8 mmol) were added to the reaction bottle and stirred, and reacted at room temperature (25 ° C) for 14 hours. The reaction solution was slowly added to 100 mL of water at room temperature to precipitate a yellow solid powder. Stirred at room temperature for 1 hour, filtered, and the filter cake was separated and purified by gradient elution of silica gel column chromatography (dichloromethane / methanol) to obtain a yellow solid crude product (0.10 g, 8%). The product was prepared and purified by HPLC [MeCN-HCOONH4 (10 mM), MeCN (0%-75%), 20 mL / min, 35 min]. The obtained product solution was extracted with dichloromethane, washed with water, dried, and the organic phase was spin-dried and freeze-dried to obtain LZ002-013 (24 mg, purity 96.88%). 1HNMR(600MHz,DMSO-d6)δ10.99(d,J=12.7Hz,2H),7.51(d,J=3.2Hz,1H),7.28(t,J=7.7Hz ,1H),7.24(dd,J=8.6,2.4Hz,1H),7.20–7.15(m,1H),7.03(d,J=2.4Hz,1H),6.97(dd,J=7. 4,0.8Hz,1H),6.78(ddd,J=15.3,14.1,8.1Hz,3H),6.10(t,J=6.2Hz,1H),5.41(s,1H),5. 14–5.08(m,2H),4.24–4.13(m,2H),4.11(dt,J=7.8,4.0Hz,1H),4.00(d,J=6.1Hz,2H),3.9 2(dt,J=10.3,3.7Hz,2H),3.81(s,3H),3.48(s,3H),3.42–3.33(m,4H),3.28(s,1H),2.94 –2.87(m,1H),2.59(t,J=7.3Hz,3H),2.48–2.41(m,2H),2.41–2.37(m,1H),2.31(qd,J=13. 2,4.5Hz,2H),2.22(d,J=11.2Hz,1H),2.14(s,2H),2.05–1.99(m,1H),1.85(ddd,J=12.5,4 .4,2.0Hz,2H),1.69–1.59(m,4H),1.49–1.42(m,2H),1.19(t,J=7.4Hz,3H).HRMS(ESI)for C 45 H 58 N 11 O7[M+H] + ,Calcd:864.4515,Found:864.4490.

[0394] Example 66: 3-((4-(4-(4-(1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)pyrrolidine-3-carbonyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LZ002-014)

[0395]

[0396] The synthesis method is the same as Example 65, yellow solid. 1H NMR(600MHz,DMSO-d6)δ10.98(d,J=35.6Hz,2H),7.51(d,J=3.3Hz,1H),7.30(t,J=7 .7Hz,1H),7.26(dd,J=8.6,2.4Hz,1H),7.19(d,J=3.3Hz,1H),7.06–6.98(m,2H),6.8 3(d,J=8.6Hz,1H),6.76(d,J=7.5Hz,1H),6.72(d,J=8.3,1H),5.10(ddd,J=13.3,5.2 ,2.2Hz,1H),4.70–4.52(m,2H),4.14–4.09(m,1H),3.94–3.87(m,2H),3.81(s,3H),3 .63(q,J=8.5Hz,1H),3.57–3.53(m,2H),3.51–3.44(m,4H),3.38–3.32(m,4H),2.96– 2.85(m,2H),2.62–2.57(m,3H),2.54(d,J=11.9Hz,2H),2.35(s,1H),2.19–2.12(m,1 H),2.07(s,1H),2.01–1.95(m,2H),1.88–1.78(m,4H),1.72–1.51(m,5H),1.50–1.38 (m,1H),1.32(d,J=22.0Hz,1H),1.27–1.22(m,3H),1.20–1.17(m,3H).HRMS(ESI)for C 45 H 60 N 11 O7[M+H] + ,Calcd:878.4672,Found:878.4608.

[0397] Example 67: 3-((4-(4-(4-(1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)piperidine-4-carbonyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LZ002-015)

[0398]

[0399] The synthesis method is the same as Example 65, yellow solid. 1H NMR (600MHz, DMSO-d6) δ11.01(s,1H),10.96(s,1H),7.51(d,J=3.2Hz,1H),7.43(t,J=7.7 Hz,1H),7.30(d,J=7.4Hz,1H),7.26(dd,J=8.6,2.3Hz,1H),7.17(dd,J=19.4,5.6Hz,2H),7 .03(d,J=2.3Hz,1H),6.82(d,J=8.6Hz,1H),6.76(d,J=7.5Hz,1H),5.11(dd,J=13.3,5.2H z,1H),4.44(d,J=17.3Hz,1H),4.31(d,J=17.3Hz,1H),4.12(dt,J=7.6,4.0Hz,1H),3.95–3 .90(m,2H),3.81(s,3H),3.52(dd,J=29.1,14.0Hz,4H),3.42–3.32(m,7H),2.95–2.88(m, 1H),2.86–2.77(m,3H),2.62–2.56(m,3H),2.55–2.51(m,3H),2.48–2.43(m,2H),2.34(tt, J=11.3,3.5Hz,1H),2.04–1.93(m,1H),1.88–1.77(m,4H),1.76–1.71(m,4H),1.64(tdd,J =15.5,9.4,4.1Hz,3H),1.57(dt,J=12.1,6.0Hz,2H),1.19(t,J=7.4Hz,3H).HRMS(ESI)for C 47 H 62 N 11 O7[M+H] + ,Calcd:892.4828,Found:892.4769.

[0400] Example 68: 3-((4-(4-(4-(2-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoquinolin-4-yl)-2-azaspiro[3.3]heptane-6-carbonyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LZ002-016)

[0401]

[0402] The synthesis method is the same as Example 65, yellow solid. 1H NMR (600MHz, DMSO-d6) δ11.00(s,1H),10.96(s,1H),7.51(d,J=3.2Hz,1H),7.30(t,J=7.7Hz,1H) ,7.25(dd,J=8.6,2.3Hz,1H),7.18(d,J=3.2Hz,1H),7.07–7.02(m,2H),6.82(d,J=8.7Hz,1H),6. 75(d,J=7.5Hz,1H),6.53(dd,J=8.0,0.9Hz,1H),5.32(m,1H),5.09(dd,J=13.3,5.2Hz,1H),4.42 (d,J=16.9Hz,1H),4.28(d,J=16.9Hz,1H),4.14–4.09(m,1H),4.02(q,J=7.2Hz,2H),3.94–3.90( m,2H),3.87(q,J=7.5Hz,2H),3.81(s,3H),3.45(t,J=4.4Hz,2H),3.36(m,4H),3.27–3.21(m,2H) ,2.94–2.89(m,1H),2.62–2.56(m,3H),2.54–2.45(m,5H),2.45–2.40(m,1H),2.39–2.35(m,4H), 2.35–2.30(m,1H),2.05–1.95(m,2H),1.85(ddd,J=12.4,4.3,1.9Hz,2H),1.79(d,J=11.9Hz,2H) ,1.64(td,J=12.0,4.2Hz,2H),1.55(dd,J=12.4,8.8Hz,2H),1.19(t,J=7.4Hz,3H).HRMS(ESI)for C 48 H 62 N 11 O7[M+H] + ,Calcd:904.4828,Found:904.4772.

[0403] Example 69: 3-[4-(4-{2-[2-(2,6-dioxy-piperidin-3-yl)-1-oxy-2,3-dihydro-1H-isoindol-4-oxy]-acetyl}-piperazin-1-yl)-3-methoxy-phenylamino]-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-021)

[0404]

[0405] Step 1: tert-Butyl 4-(2-methoxy-4-nitrophenyl)piperazine-1-carboxylate

[0406] Disperse 1-tert-Butyloxycarbonylpiperazine (10.00 g, 53.76 mmol) and 2-fluoro-5-nitroanisole (11.03 g, 64.51 mmol) in DMF (100 mL), add DIPEA (20.81 g, 161.28 mmol), and stir at 90 ° C overnight. After the reaction is completed by TLC monitoring, the reaction system is diluted with water (500 mL), extracted with ethyl acetate three times, the organic phases are combined and washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (PE: EA = 2: 1) to obtain a brown solid product (9.50 g, 52.49%). MS (ESI), m / z: 338.2 [M + H] + .

[0407] Step 2: tert-Butyl 4-(4-amino-2-methoxyphenyl)piperazine-1-carboxylate

[0408] The product of step 1 (9.50 g, 28.19 mmol) was added to a 250 mL single-mouth bottle, and ethanol (100 mL) and H2O (50 mL) were added. Ammonium chloride (7.47 g, 140.95 mmol) and iron powder (7.89 g, 140.95 mmol) were added under stirring at room temperature, and stirred at 100°C for 4 hours. Filtered with diatomaceous earth, the filter cake was washed with methanol. The organic phase was dried and separated by silica gel column chromatography (PE: EA = 3: 97) to obtain a brown-yellow solid product (4.50 g, 52.02%). MS (ESI), m / z: 308.4 [M+H] + .

[0409] Step 3: tert-Butyl 4-(4-((3-cyano-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazine-1-carboxylate

[0410] 2-12 (3.0 g, 10 mmol) was dispersed in dry 1,4-dioxane (60 mL), and the product of step 2 (3.07 g, 10 mmol), palladium acetate (224 mg, 1.0 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (1.16 g, 2 mmol) and potassium carbonate (2.06 g, 15 mmol) were added to the reaction system. After inert gas replacement, the reaction system was stirred at 90 ° C overnight. After the reaction was completed by TLC monitoring, the system was cooled to room temperature, diluted with water, extracted with ethyl acetate, the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (DCM: MeOH = 80: 1) to obtain a reddish brown solid product (3.10 g, 57%). MS (ESI), m / z: 538.2 [M + H] + .

[0411] Step 4: tert-Butyl 4-(4-((3-carbamoyl-5-ethyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrazin-2-yl)amino)-2-methoxyphenyl)piperazine-1-carboxylate

[0412] Disperse the product of step 3 (3.10 g, 5.77 mmol) into a mixed solution of DMSO (25 mL) / ethanol (50 mL), add 4N sodium hydroxide (10 mL, 40 mmol), slowly add 30% hydrogen peroxide (5 mL, 52.5 mmol), and keep the temperature below 20°C. After the addition is complete, warm to room temperature and continue stirring for 2 hours. After the reaction is completed by TLC monitoring, the reaction system is diluted with water, extracted with ethyl acetate, the organic phases are combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product (3.10 g, 97%). MS (ESI), m / z: 556.3 [M+H] + .

[0413] Step 5: 6-ethyl-3-((3-methoxy-4-(piperazin-1-yl)phenyl)amino)-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide

[0414] The product of step 4 (3.10 g, 5.58 mmol) was dissolved in a mixed solvent of dichloromethane / trifluoroacetic acid (2:1), stirred at room temperature for 3 hours, concentrated under reduced pressure after the reaction was complete, and column chromatography (DCM:MeOH=10:1) gave a yellow solid (2.34 g, 92%). MS (ESI), m / z: 456.3 [M+H] + .

[0415] Step 6: 3-[4-(4-{2-[2-(2,6-dioxy-piperidin-3-yl)-1-oxy-2,3-dihydro-1H-isoindol-4-yloxy]-acetyl}-piperazin-1-yl)-3-methoxy-phenylamino]-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-021)

[0416] The product of step 5 (1.63g, 3.58mmol), 2-40 (0.88g, 2.75mmol), DMF (40mL) and DIPEA (1.07g, 8.25mmol) were added to a 100mL single-mouth bottle and stirred at room temperature for 30 minutes. HATU (1.25g, 3.30mmol) was added again and stirred at room temperature overnight. After the reaction was completed by TLC detection, water was slowly added dropwise to the system, and a yellow solid powder was precipitated. After stirring, it was quickly filtered and the filter cake was naturally air-dried. Silica gel column chromatography was separated and purified (DCM: MeOH = 85: 15) to obtain a yellow solid crude product (1.00g, 50%). HPLC preparation and purification [MeCN-HCOONH4 (10mM), MeCN (0%-75%), 20mL / min, 35min]. The obtained finished product solution was extracted with dichloromethane, washed with water, dried, the organic phase was spin-dried, and freeze-dried to obtain 124 mg of pure LZ002-021. 1 H NMR (600MHz, DMSO) δ11.04(s,1H),10.96(s,1H),7.53(d,J=2.6Hz,1H),7.47(t,J=7.8Hz,1H),7.34–7.30(m,2H),7.21–7.16(m,2H),7.05(d,J=2. 3Hz,1H),6.84(d,J=8.6Hz,1H),6.77(d,J=7.5Hz,1H),5.11(dd,J=13.3,5.1Hz,1H),5.06(s,2H),4.42(d,J=17.3Hz,1H),4.30(d,J=17.3Hz,1H), 4.15–4.08(m,1H),3.95–3.89(m,2H),3.83(s,3H),3.64–3.56(m,4H),3. 37(td,J=11.8,2.1Hz,2H),3.02–2.95(m,2H),2.93–2.90(m,3H),2.62–2 .55(m,3H),2.45(td,J=13.4,4.6Hz,1H),2.05–1.96(m,1H),1.88–1.83(m,2H),1.64(qd,J=12.3,4.5Hz,2H),1.19(t,J=7.4Hz,3H).HRMS(ESI)for C 38 H 46 N9O8[M+H] + ,Calcd:756.3464,Found:756.3381.

[0417] Example 70: 3-[4-(4-{2-[2-(2,6-dioxy-piperidin-3-yl)-1-oxy-2,3-dihydro-1H-isoindol-4-ylamino]-acetyl}-piperazin-1-yl)-3-methoxy-phenylamino]-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-022)

[0418]

[0419] The synthesis method is the same as Example 69, yellow solid. 1 H NMR (600MHz, DMSO) δ11.04 (s, 1H), 10.96 (s, 1H), 7.53 (d, J = 2.6Hz, 1H), 7.34 –7.28(m,2H),7.20(d,J=3.3Hz,1H),7.05(d,J=2.3Hz,1H),7.00(d,J=7.4Hz, 1H),6.84(d,J=8.6Hz,1H),6.80(d,J=8.1Hz,1H),6.77(d,J=7.5Hz,1H),5.5 9(t,J=4.8Hz,1H),5.11(dd,J=13.3,5.1Hz,1H),4.32(d,J=16.9Hz,1H),4.20 (d,J=16.9Hz,1H),4.15–4.08(m,3H),3.95–3.89(m,2H),3.83(s,3H),3.70– 3.60(m,4H),3.37(td,J=11.8,2.1Hz,2H),3.01–2.87(m,5H),2.65–2.56(m,3 H),2.38(qd,J=13.2,4.5Hz,1H),2.03(dtd,J=12.8,5.4,2.3Hz,1H),1.88–1. 83(m,2H),1.64(qd,J=12.1,4.5Hz,2H),1.19(t,J=7.4Hz,3H).HRMS(ESI)for C 38 H 47 N 10 O7[M+H] + ,Calcd:755.3624,Found:755.3550.

[0420] Example 71: 3-{4-[4-(6-{2-[2-(2,6-dioxy-piperidin-3-yl)-1-oxy-2,3-dihydro-1H-isoindol-4-yloxy]-acetyl}-2,6-diaza-spiro[3.3]hept-2-yl)-piperidin-1-yl]-3-methoxy-phenylamino}-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-023)

[0421]

[0422] Step 1: tert-Butyl 6-(1-(((9H-fluoren-9-yl)methoxy)carbonyl)piperidin-4-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate

[0423] Disperse Fmoc-4-piperidone (500 mg, 1.55 mmol) and 2-Boc-2,6-diazaspiro[3.3]heptane hemioxalate (465.59 mg, 1.55 mmol) in DCM (40 mL), add acetic acid (2 mL), and stir at room temperature for 1 hour. Add sodium acetate borohydride (990.35 mg, 4.67 mmol) and stir at room temperature overnight. After concentration under reduced pressure, separate by column chromatography (PE:EA=1:1) to obtain a colorless oil (700 mg, 89.74%). MS (ESI), m / z: 504.2[M+H] + .

[0424] Step 2: tert-Butyl 6-(piperidin-4-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate

[0425] Add the product of step 1 (700 mg, 1.39 mmol), THF (20 mL) and tetrabutylammonium fluoride (4 mL 1M, 4.17 mmol) to a 100 mL single-necked bottle and stir at room temperature overnight. Concentrate under reduced pressure and set aside. MS (ESI), m / z: 282.4 [M+H] + .

[0426] Step 3: 3-{4-[4-(6-{2-[2-(2,6-dioxy-piperidin-3-yl)-1-oxy-2,3-dihydro-1H-isoindol-4-yloxy]-acetyl}-2,6-diaza-spiro[3.3]hept-2-yl)-piperidin-1-yl]-3-methoxy-phenylamino}-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-023)

[0427] The synthesis method is the same as Example 69, yellow solid. 1H NMR (600MHz, DMSO) δ11.01 (s, 1H), 10.97 (s, 1H), 7.52 (d, J = 2.5Hz, 1H), 7.48 (t, J = 7. 8Hz,1H),7.35(d,J=7.3Hz,1H),7.25(dd,J=8.6,2.3Hz,1H),7.19(d,J=2.6Hz,1H),7. 15(d,J=8.1Hz,1H),7.04(d,J=2.3Hz,1H),6.82(d,J=8.7Hz,1H),6.76(d,J=7.5Hz,1H ),5.11(dd,J=13.3,5.1Hz,1H),4.75(s,2H),4.43(d,J=17.3Hz,1H),4.33–4.27(m,3H ),4.33–4.27(m,3H),4.17–4.07(m,1H),4.00(s,2H),3.95–3.90(m,2H),3.80(s,3H) ,3.37(td,J=11.8,2.1Hz,2H),3.29–3.17(m,6H),2.92(ddd,J=17.3,13.6,5.4Hz,1H) ,2.63–2.53(m,5H),2.50–2.43(m,1H),2.07–1.99(m,2H),1.89–1.83(m,2H),1.72–1. 66(m,2H),1.66–1.60(m,2H),1.33–1.26(m,2H),1.20(t,J=7.4Hz,3H).HRMS(ESI)for C 44 H 55 N 10 O8[M+H] + ,Calcd:851.4199,Found:851.4163.

[0428] Example 72: 3-{4-[4-(6-{2-[2-(2,6-dioxy-piperidin-3-yl)-1-oxy-2,3-dihydro-1H-isoindol-4-ylamino]-acetyl}-2,6-diaza-spiro[3.3]hept-2-yl)-piperidin-1-yl]-3-methoxy-phenylamino}-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-024)

[0429]

[0430] The synthesis method is the same as Example 71, yellow solid. 1H NMR (600MHz, DMSO-d6) δ11.01(s,1H),10.98(s,1H),7.52(d,J=3.3Hz,1H),7.29(t,J=7.7 Hz,1H),7.25(dd,J=8.6,2.4Hz,1H),7.19(d,J=3.3Hz,1H),7.04(d,J=2.3Hz,1H),6.98(d ,J=7.4Hz,1H),6.82(d,J=8.6Hz,1H),6.76(d,J=7.5Hz,1H),6.66(d,J=8.1Hz,1H),5.74( t,J=5.8Hz,1H),5.11(dd,J=13.3,5.1Hz,1H),4.33–4.25(m,3H),4.17(d,J=17.0Hz,1H),4 .15–4.07(m,1H),3.99(s,2H),3.95–3.89(m,2H),3.81(s,1H),3.80(s,3H),3.44–3.32(m ,6H),3.29–3.18(m,4H),2.92(ddd,J=17.4,13.6,5.5Hz,1H),2.66–2.51(m,5H),2.34(qd, J=13.2,4.5Hz,1H),2.07–2.01(m,1H),1.85(ddd,J=12.5,4.4,2.0Hz,2H),1.77–1.68(m, 2H),1.63(qd,J=12.3,4.4Hz,2H),1.36–1.28(m,2H),1.19(t,J=7.4Hz,3H).HRMS(ESI)for C 44 H 56 N 11 O7[M+H] + ,Calcd:850.4359,Found:850.4263.

[0431] Example 73: 3-{4-[4-(5-{2-[2-(2,6-dioxy-piperidin-3-yl)-1-oxy-2,3-dihydro-1H-isoindol-4-yloxy]-acetyl}-hexahydropyrrolo[3,4-c]pyrrol-2-yl)-piperidin-1-yl]-3-methoxy-phenylamino}-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-025)

[0432]

[0433] The synthesis method is the same as Example 71, yellow solid. 1H NMR (600MHz, DMSO-d6) δ11.01(s,1H),10.97(d,J=3.4Hz,1H),7.52(d,J=3.3Hz,1H),7. 46(t,J=7.8Hz,1H),7.31(d,J=7.5Hz,1H),7.25(dt,J=8.6,1.7Hz,1H),7.20(d,J=3.3Hz ,1H),7.14(d,J=8.2Hz,1H),7.04(d,J=2.4Hz,1H),6.81(d,J=8.6Hz,1H),6.76(d,J=7.5 Hz,1H),5.11(ddd,J=13.3,5.2,2.0Hz,1H),4.95–4.85(m,2H),4.40(dd,J=17.3,3.0Hz, 1H),4.30–4.24(m,1H),4.16–4.07(m,1H),3.94–3.89(m,2H),3.81(s,3H),3.72–3.65( m,1H),3.59–3.51(m,1H),3.43–3.32(m,4H),3.31–3.22(m,3H),2.96–2.81(m,2H),2.76 –2.68(m,1H),2.63–2.50(m,7H),2.50–2.40(m,2H),2.06–1.96(m,2H),1.89–1.80(m,4H ),1.63(qd,J=12.2,4.4Hz,2H),1.56–1.45(m,2H),1.19(t,J=7.4Hz,3H).HRMS(ESI)for C 45 H 57 N 10 O8[M+H] + ,Calcd:865.4355,Found:865.4392.

[0434] Example 74: 3-{4-[4-(5-{2-[2-(2,6-dioxy-piperidin-3-yl)-1-oxy-2,3-dihydro-1H-isoindol-4-ylamino]-acetyl}-hexahydropyrrolo[3,4-c]pyrrol-2-yl)-piperidin-1-yl]-3-methoxy-phenylamino}-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-026)

[0435]

[0436] The synthesis method is the same as Example 71, yellow solid. 1H NMR (600MHz, DMSO-d6) δ11.01(s,1H),10.99(s,1H),7.52(d,J=3.2Hz,1H),7.29(t,J=7.7Hz,1H ),7.24(dt,J=8.6,2.3Hz,1H),7.21(d,J=3.2Hz,1H),7.04(t,J=2.6Hz,1H),6.98(d,J=7.4Hz,1 H),6.81(dd,J=8.6,2.3Hz,1H),6.77(d,J=7.5Hz,1H),6.73(d,J=8.1Hz,1H),5.64–5.59(m,1H) ,5.11(dd,J=13.3,5.1Hz,1H),4.29(d,J=16.9Hz,1H),4.18(dd,J=16.9,2.5Hz,1H),4.15–4.07 (m,1H),4.02–3.93(m,2H),3.94–3.88(m,2H),3.80(s,3H),3.73(ddd,J=11.4,8.4,3.3Hz,1H), 3.61–3.53(m,1H),3.48–3.41(m,1H),3.39–3.32(m,2H),3.29–3.21(m,3H),2.96–2.83(m,2H), 2.77–2.70(m,1H),2.66–2.44(m,9H),2.37(qt,J=13.3,3.9Hz,1H),2.07–1.99(m,2H),1.89–1. 80(m,4H),1.63(qd,J=12.2,4.5Hz,2H),1.57–1.47(m,2H),1.19(t,J=7.3Hz,3H).HRMS(ESI)for C 45 H 58 N 11 O7[M+H] + ,Calcd:864.4515,Found:864.4492.

[0437] Example 75: 3-{4-[4-(5-{2-[2-(2,6-dioxopiperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-4-yloxy]-acetyl}-2,5-diazabicyclo[2.2.2]octyl-2-yl)-piperidin-1-yl]-3-methoxyphenylamino}-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-027)

[0438]

[0439] The synthesis method is the same as Example 71, yellow solid.1 H NMR(600MHz,DMSO-d6)δ11.00(d,J=2.4Hz,1H),10.97(s,1H),7.52(d,J=3.3Hz,1H), 7.46(q,J=7.6Hz,1H),7.33(dd,J=7.5,6.0Hz,1H),7.27–7.22(m,1H),7.21–7.15(m, 2H),7.07–7.03(m,1H),6.82(dd,J=8.6,4.8Hz,1H),6.76(d,J=7.4Hz,1H),5.11(ddd ,J=13.3,5.3,1.7Hz,1H),5.00–4.89(m,2H),4.40(dd,J=17.3,4.1Hz,1H),4.32–4.25 (m,1H),4.16–4.07(m,1H),3.97–3.89(m,3H),3.81(d,J=2.5Hz,3H),3.54–3.48(m,1 H),3.37(td,J=11.8,2.0Hz,2H),3.29–3.19(m,4H),2.95–2.84(m,2H),2.63–2.52(m ,5H),2.48–2.40(m,2H),2.04–1.98(m,1H),1.97–1.82(m,6H),1.80–1.69(m,2H),1. 68–1.54(m,3H),1.51–1.41(m,2H),1.19(t,J=7.4Hz,3H).MS(ESI),m / z:865.43[M+H] + .

[0440] Example 76: 3-{4-[4-(5-{2-[2-(2,6-dioxopiperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-4-ylamino]-acetyl}-2,5-diazabicyclo[2.2.2]octyl-2-yl)-piperidin-1-yl]-3-methoxyphenylamino}-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-028)

[0441]

[0442] The synthesis method is the same as Example 71, yellow solid. 1H NMR (600MHz, DMSO-d6) δ11.00(s,1H),10.98(s,1H),7.52(d,J=3.3Hz,1H),7.30(t,J=7.7Hz,1H ),7.25(dd,J=8.6,2.3Hz,1H),7.19(d,J=3.3Hz,1H),7.06–7.03(m,1H),6.99(d,J=7.4Hz,1H),6 .82(dd,J=8.6,4.7Hz,1H),6.80–6.74(m,2H),5.59–5.51(m,1H),5.11(dd,J=13.2,5.2Hz,1H),4 .40–4.28(m,2H),4.23–4.16(m,1H),4.11(dtt,J=11.2,7.8,3.9Hz,1H),4.08–3.95(m,2H),3.95 –3.90(m,2H),3.81(d,J=3.0Hz,3H),3.57–3.50(m,1H),3.36(td,J=11.8,2.0Hz,2H),3.31–3.20 (m,4H),2.97–2.88(m,2H),2.65–2.51(m,5H),2.48–2.43(m,1H),2.38(qd,J=13.3,4.6Hz,1H),2 .07–2.00(m,1H),1.99–1.88(m,3H),1.88–1.83(m,3H),1.81–1.68(m,2H),1.63(qd,J=12.2,4.4 Hz,2H),1.59–1.52(m,1H),1.51–1.42(m,2H),1.19(t,J=7.4Hz,3H).MS(ESI),m / z:864.44[M+H] + .

[0443] Example 77: 3-{4-[4-(8-{2-[2-(2,6-dioxopiperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-4-yloxy]-acetyl}-3,8-diazabicyclo[3.2.1]octyl-3-yl)-piperidin-1-yl]-3-methoxyphenylamino}-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-029)

[0444]

[0445] The synthesis method is the same as Example 71, yellow solid. 1H NMR(600MHz,DMSO-d6)δ11.00(s,1H),10.96(s,1H),7.51(d,J=3.3Hz,1H),7.50–7.45(m,1H),7.3 3(dd,J=7.5,0.7Hz,1H),7.25(dd,J=8.8,2.5Hz,1H),7.20–7.15(m,2H),7.03(d,J=2.3Hz,1H),6. 82(d,J=8.6Hz,1H),6.75(d,J=7.5Hz,1H),5.11(ddd,J=13.3,5.3,3.1Hz,1H),4.99(d,J=14.7Hz, 1H),4.90(dd,J=14.6,3.4Hz,1H),4.46(d,J=6.8Hz,1H),4.41(dd,J=17.2,1.6Hz,1H),4.34(s,1H ),4.28(d,J=17.2Hz,1H),4.16–4.07(m,1H),3.95–3.89(m,2H),3.81(s,3H),3.36(td,J=11.8,2. 0Hz,2H),3.32–3.27(m,2H),2.96–2.87(m,1H),2.78–2.70(m,2H),2.62–2.55(m,3H),2.55–2.51( m,1H),2.49–2.43(m,1H),2.43–2.38(m,1H),2.31–2.25(m,2H),2.05–1.95(m,2H),1.89–1.82(m, 4H),1.81–1.71(m,3H),1.69–1.59(m,3H),1.59–1.50(m,2H),1.19(t,J=7.4Hz,3H).HRMS(ESI)for C 45 H 57 N 10 O8[M+H] + ,Calcd:865.4355,Found:865.4317.

[0446] Example 78: 3-{4-[4-(8-{2-[2-(2,6-dioxopiperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-4-ylamino]-acetyl}-3,8-diazabicyclo[3.2.1]oct-3-yl)-piperidin-1-yl]-3-methoxyphenylamino}-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-030)

[0447]

[0448] The synthesis method is the same as Example 71, yellow solid. 1 H NMR (600MHz, DMSO-d6) δ11.00(s,1H),10.98(s,1H),7.51(d,J=3.3Hz,1H),7.30(t,J=7.7Hz,1H),7.24 (dd,J=8.6,2.4Hz,1H),7.19(d,J=3.2Hz,1H),7.03(d,J=2.3Hz,1H),6.99(dd,J=7.5,0.8Hz,1H),6.82 (d,J=8.6Hz,1H),6.77(d,J=8.1Hz,1H),6.75(d,J=7.6Hz,1H),5.65–5.61(m,1H),5.11(dd,J=13.3,5. 1Hz,1H),4.48(d,J=6.8Hz,1H),4.42(s,1H),4.31(d,J=16.9Hz,1H),4.19(dd,J=16.9,1.9Hz,1H),4.15 –4.06(m,2H),3.97(t,J=4.9Hz,1H),3.95–3.89(m,2H),3.81(s,3H),3.36(td,J=11.8,2.0Hz,2H),3.3 3–3.27(m,2H),2.92(ddd,J=17.0,13.5,5.3Hz,1H),2.77(d,J=10.4Hz,1H),2.72(d,J=10.7Hz,1H),2.6 5–2.56(m,3H),2.55–2.51(m,2H),2.43–2.33(m,2H),2.32–2.25(m,2H),2.06–1.96(m,2H),1.90–1.82 (m,4H),1.82–1.72(m,2H),1.70–1.59(m,3H),1.58–1.51(m,2H),1.19(t,J=7.4Hz,3H).HRMS(ESI)forC 45 H 58 N 11 O7[M+H] + ,Calcd:864.4515,Found:864.4425.

[0449] Example 79: 3-{4-[4-(3-{2-[2-(2,6-dioxopiperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-4-yloxy]-acetyl}-3,8-diazabicyclo[3.2.1]octyl-8-yl)-piperidin-1-yl]-3-methoxyphenylamino}-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-031)

[0450]

[0451] The synthesis method is the same as Example 71, yellow solid. 1 H NMR (600MHz, DMSO-d6) δ11.00(s,1H),10.96(s,1H),7.51(d,J=3.3Hz,1H),7.46(t,J=7.8Hz,1H),7 .32(d,J=7.2Hz,1H),7.25(dd,J=8.6,2.3Hz,1H),7.19(d,J=3.3Hz,1H),7.14(ddd,J=8.2,1.7,0.8 Hz,1H),7.05(d,J=2.4Hz,1H),6.82(d,J=8.6Hz,1H),6.76(d,J=7.5Hz,1H),5.10(td,J=14.1,13.7 ,5.4Hz,2H),4.86(dd,J=14.8,2.8Hz,1H),4.40(d,J=17.3Hz,1H),4.28(d,J=17.3Hz,1H),4.16–4. 07(m,1H),3.95–3.89(m,2H),3.84(d,J=12.6Hz,1H),3.82(s,3H),3.54–3.48(m,2H),3.44(d,J=12 .2Hz,1H),3.40–3.30(m,3H),2.96–2.87(m,1H),2.83(d,J=12.5Hz,1H),2.63–2.50(m,5H),2.49–2 .41(m,1H),2.39–2.33(m,1H),2.05–1.96(m,2H),1.98–1.91(m,2H),1.85(ddd,J=12.6,4.4,2.0Hz ,2H),1.82–1.75(m,2H),1.70–1.59(m,3H),1.48–1.37(m,4H),1.19(t,J=7.4Hz,3H).HRMS(ESI)for C 45 H 57 N 10 O8[M+H] + ,Calcd:865.4355,Found:865.4296.

[0452] Example 80: 3-{4-[4-(3-{2-[2-(2,6-dioxopiperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-4-ylamino]-acetyl}-3,8-diazabicyclo[3.2.1]octyl-8-yl)-piperidin-1-yl]-3-methoxyphenylamino}-6-ethyl-5-(tetrahydropyran-4-ylamino)-pyrazine-2-carboxylic acid amide (LZ002-032)

[0453]

[0454] The synthesis method is the same as Example 71, yellow solid. 1 H NMR (600MHz, DMSO-d6) δ11.00(s,1H),10.99(s,1H),7.52(d,J=3.3Hz,1H),7.29(t,J=7. 7Hz,1H),7.25(dd,J=8.6,2.3Hz,1H),7.20(d,J=3.3Hz,1H),7.05(d,J=2.3Hz,1H),6.99( d,J=7.4Hz,1H),6.82(d,J=8.6Hz,1H),6.77(dd,J=7.3,2.1Hz,2H),5.55(q,J=5.4Hz,1H ),5.14–5.08(m,1H),4.31(dd,J=16.9,3.0Hz,1H),4.18(d,J=16.9Hz,1H),4.18–4.07(m, 2H),3.95–3.84(m,4H),3.81(s,3H),3.59–3.47(m,3H),3.40–3.32(m,3H),3.31–3.27(m ,2H),2.92(ddd,J=17.4,13.6,5.5Hz,1H),2.85(d,J=12.3Hz,1H),2.65–2.51(m,5H),2.4 3–2.33(m,2H),2.07–1.99(m,1H),1.98–1.91(m,2H),1.88–1.83(m,2H),1.82–1.74(m,2 H),1.63(qd,J=12.2,4.4Hz,3H),1.48–1.37(m,3H),1.19(t,J=7.4Hz,3H).HRMS(ESI)for C 45 H 58 N 11 O7[M+H] + ,Calcd:864.4515,Found:864.4514.

[0455] Example 81: 3-((4-(1-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)acetyl)piperidin-4-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LZ002-033)

[0456]

[0457] Step 1: tert-Butyl 4-(2-methoxy-4-nitrophenyl)-3,6-dihydropyridine-1(2H)-carboxylate

[0458] 2-Bromo-5-nitroanisole (0.51 g, 2.2 mmol), N-Boc-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (0.74 g, 2.4 mmol), Pd(dppf)Cl2 (0.09 g), anhydrous potassium carbonate (1.22 g, 8.8 mmol) and dioxane (24 mL) were added to a 50 mL reaction bottle, and purified water (8.0 mL) was added after stirring evenly. After N2 replacement, the temperature was raised to 80 ° C and reacted for 18 hours. After TLC monitoring, the reaction solution was poured into purified water, extracted with ethyl acetate three times, and the organic phases were combined and dried with anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography (PE:EA=10:1→4.5:1) to obtain 0.9 g of a yellow transparent oil.

[0459] Step 2: tert-Butyl 4-(4-amino-2-methoxyphenyl)piperidine-1-carboxylate

[0460] Add the product of step 1 (0.3g, 0.7mmol), ethanol (4.0mL, 12.5v / m), Pd / C (10%, 0.1g) and ammonium formate (0.25g, 5.0eq) into a 25mL reaction bottle, stir and replace with N2, heat to 55℃ and react for 2-3 hours, add 1.4mmol (0.6g) of the product of step 1, and continue to react for 2 hours (45℃). After the reaction is complete, combine the reaction solutions, add magnesium sulfate, stir and dry, filter and concentrate to obtain a pink solid (0.6g, 89.6%).

[0461] Step 3: 3-((4-(1-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)acetyl)piperidin-4-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LZ002-033)

[0462] The synthesis method is the same as Example 71, yellow solid. 1H NMR (600MHz, DMSO-d6) δ11.16(s,1H),10.96(s,1H),7.55(d,J=2.5Hz,1H),7.47(t,J=7.8Hz,1H),7.37(d,J=8. 3Hz,1H),7.33(d,J=7.4Hz,1H),7.23(d,J=2.5Hz,1H),7.18(d,J=8.2Hz,1H),7.05(d,J=8.2Hz,1H),7.02(s,1H) ,6.79(d,J=7.5Hz,1H),5.11(dd,J=13.3,5.1Hz,1H),5.05(d,J=3.2Hz,2H),4.47(d,J=12.1Hz,1H),4.41(d,J=1 7.3Hz,1H),4.29(d,J=17.4Hz,1H),4.19–4.09(m,1H),3.98–3.89(m,3H),3.83(s,3H),3.40–3.33(m,2H),3.20–

[0463] 3.13(m,1H),3.09(tt,J=12.1,3.3Hz,1H),2.96–2.87(m,1H),2.71–2.6 4(m,1H),2.59(q,J=7.3Hz,3H),2.49–2.41(m,1H),2.01(ddd,J=12.4,6. 2,2.1Hz,1H),1.86(ddd,J=12.3,4.4,2.0Hz,2H),1.78–1.71(m,2H),1.70–1.60(m,3H),1.49–1.39(m,1H),1.20(t,J=7.4Hz,3H).HRMS(ESI)for C 39 H 47 N8O8[M+H] + ,Calcd:755.3511,Found:755.3505.

[0464] Example 82: 3-((4-(1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-4-yl)glycylamino)piperidin-4-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (LZ002-034)

[0465] The synthesis method is the same as Example 81, yellow solid. 1H NMR(600MHz,DMSO-d6)δ11.16(s,1H),10.98(s,1H),7.56–7.53(m,1H),7.38(dd,J=8.3,2 .1Hz,1H),7.31(t,J=7.7Hz,1H),7.25–7.21(m,1H),7.05(d,J=8.4Hz,1H),7.01(s,1H),7 .00(d,J=7.3Hz,1H),6.82(d,J=8.1Hz,1H),6.79(d,J=7.5Hz,1H),5.54(t,J=5.1Hz,1H), 5.12(dd,J=13.2,5.1Hz,1H),4.54(d,J=12.2Hz,1H),4.32(dd,J=16.9,4.1Hz,1H),4.20(d d,J=16.9,6.5Hz,1H),4.18–4.04(m,4H),3.94–3.89(m,2H),3.83(s,3H),3.40–3.33(m,2 H),3.18–3.06(m,2H),2.97–2.88(m,1H),2.73–2.66(m,1H),2.65–2.56(m,3H),2.39(qd, J=13.3,4.5Hz,1H),2.03(dtd,J=12.5,5.1,2.6Hz,1H),1.89–1.83(m,2H),1.79–1.72(m, 2H),1.64(qd,J=12.1,4.5Hz,3H),1.49–1.39(m,1H),1.20(t,J=7.3Hz,3H).HRMS(ESI)for C 39 H 48 N9O7[M+H] + ,Calcd:754.3671,Found:754.3682.

[0466] Example 83: 3-((4-(4-(-4-((1-(2-(2,6-dioxopiperidin-3-yl)-7-fluoro-1,3-dioxoisoindol-4-yl)azetidin-3-yl)methyl)piperazin-1-yl)piperidin-1-yl)-3-methoxyphenyl)amino)-6-ethyl-5-((tetrahydro-2H-pyran-4-yl)amino)pyrazine-2-carboxamide (ZLC5-69)

[0467]

[0468] Step 1:

[0469] 4,7-difluoroisobenzofuran-1,3-dione (250 mg, 1.35 mmol), piperidine-4-carboxaldehyde hydrochloride (267 mg, 1.63 mmol), potassium acetate (410 mg, 4.2 mmol) were added to the Shrek bottle in sequence, argon was evacuated 3 times, acetic acid (3 mL) was added, and the temperature was raised to 120 ° C. After 12 h, the reaction was completed, cooled to room temperature, concentrated under pressure, EA (20 mL) was added, ultrasonicated for 10 min, filtered, washed three times with PE / EA system (PE: EA = 1: 1), and dried to obtain intermediate ZLC5-61 (purple solid, 374 mg).

[0470] 1 H NMR(500MHz,DMSO-d6)δ11.10(s,1H),7.82(t,J=5.7Hz,2H),5.15(dd,J=13.0,5 .4Hz,1H),2.93–2.83(m,1H),2.64–2.57(m,1H),2.46(m,1H),2.09–2.01(m,1H).

[0471] Step 2:

[0472] To the anhydrous dichloromethane system (2mL) of the intermediate ZLC4-40 (47mg, 0.077mmol), trifluoroacetic acid (1mL) was added dropwise, stirred at room temperature overnight, concentrated under reduced pressure, and then DMF (1mL), DIPEA (0.1mL), and the intermediate ZLC5-61 (25mg, 0.085mmol) were added in sequence, and the temperature was raised to 90°C. After the reaction was completed by TLC monitoring, it was concentrated under reduced pressure and separated by normal phase column chromatography to obtain ZLC5-69 (yellow solid, 22mg, 33%)

[0473] 11H NMR (500 MHz, DMSO-d6) δ 11.09 (s, 1H), 11.02 (s, 1H), 7.53 (s, 1H), 7.47 (t, J = 8.7 Hz, 1H), 7.26 (d, J = 7.6 Hz, 1H), 7.22 (s, 1H), 7.02 (s, 1H), 6.82 (td, J = 15.1, 13.8, 8.6 Hz, 3H), 5.04 (dd, J = 12.8, 5.4 Hz, 1H), 4.26 (d, J = 5.2 Hz, 2H), 4.11 (m, 1H), 3.92 (d, J = 8.7 Hz, 2H), 3.81 (s, 5H), 3.52–3.43 (m, 2H), 2.92–2.82 (m, 2H), 2.70–2.50 (m, 11H), 2.49–2.42 (m, 3H), 1.99 (m, 2H), 1.85 (d, J = 11.3 Hz, 4H), 1.63 (qd, J = 11.6, 3.3 Hz, 4H), 1.23 (s, 4H), 1.18 (t, J = 7.3 Hz, 3H). MS (ESI), m / z: 882.4 [M+H] + .

[0474] Example 84:

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[0587] Example 85: Screening of compounds for their ability to degrade FLT3 in tumor cells

[0588] Experimental method: The MV4-11 cell line with FLT3-ITD mutation was incubated for 24 hours at a concentration of 10 nM. The optimal compounds with good degradation efficiency were obtained efficiently and quickly through immunoblotting analysis, and different concentrations were set for degradation efficiency detection. ImageJ software was used to quantify the gray value of the immunoblot band, and GraphPadPrism software was used to fit the half-degradation concentration DC of the compound. 50 value.

[0589] Table 1 and Table 2 summarize the Figure 1 and Figure 2 The quantitative analysis results showed that among the above compounds, 2-7a, 2-9a and 2-9d could significantly induce the degradation of FLT3 protein in MV4-11 cells at low nanomolar concentrations. Through the concentration-dependent degradation study, it was found that 2-7a and 2-9a had a better half-degradation concentration DC 50 The degradation study of LZ002 series molecules showed that compounds 023, 024, 025, 026, 027, 028, 031 and 032 can effectively degrade FLT3 protein in MV4-11 cells, and the DC 50 value.

[0590] Table 1. Expression levels of FLT3 protein after treatment with degradation agents

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[0595] * The degradation screening was performed three times independently, and the FLT3 Level% value is the average gray value of the FLT3 protein band in the three experiments.

[0596] Table 2. Summary of degradation agent activities

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[0599] *Activity data are based on three independent replicates of the experiment, with a half-degradation concentration of DC 50 Take the average of three data.

[0600] Example 86: In vitro tumor cell proliferation inhibitory activity

[0601] The inhibitory effect of the compound on cell proliferation was detected using CCK-8 cell counting kit (Bimake). The specific steps are as follows:

[0602] 1) Cell inoculation: MV4-11 suspension tumor cells in the logarithmic growth phase were inoculated in 96-well plates at the same density (15,000 cells / 100 μL / well).

[0603] 2) Preparation of working solution: Use the corresponding culture medium required for cell culture as the diluent (with or without solvent DMSO) to dilute the test compound and control compound stock solutions to obtain a series of working solutions with the required concentration being 3 times the final concentration. The content of solvent DMSO in each concentration group is consistent with that in the solvent control group.

[0604] 3) Co-incubation: 24 hours after inoculation, add 100 μL / well of the compound series concentration working solution to the 96-well plate, mix well and co-culture for 72 hours. All groups have at least 3 replicates, and each compound has 11 concentrations. Blank control group: only add culture medium, no cells and drugs, to eliminate the interference of culture medium on contrast color.

[0605] 4) Absorbance measurement: 10 μL of CCK-8 solution was added to each well of a 96-well plate. After incubation for 2 hours, the plate was shaken thoroughly to make it uniform. The absorbance values ​​(A) at 450 and 650 nm were measured on an ELISA reader. 450 and A 650 ).

[0606] 5) Data processing: The obtained A 450 -A 650 The original data were used to obtain the cell viability of each treatment well (calculation method is as follows); then the cell viability data and its corresponding compound concentration were used to calculate the IC of the compound on different cells using the nonlinear regression model of GraphPad Prism 7.0Demo software 50 value.

[0607] Calculation of cell survival rate: Cell survival rate (%) = [(As-Ac) / (Ab-Ac)]×100%] (As: experimental well; Ab: vehicle control well; Ac: blank well).

[0608] The results are shown in Table 3 below, where each compound has good activity against MV4-11, and the half-maximal inhibitory concentration of most compounds against MV4-11 is less than 10 nM.

[0609] Table 3. Antiproliferation activity of compounds on tumor cells

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[0619] *Activity data are based on three independent replicates of the experiment, and the half inhibitory concentration (IC) 50 Take the average of three data.

[0620] Example 87: Effect of Compound 2-7aa on Selective Degradation of FLT3, AXL, ALK and LTK in Tumor Cells

[0621] Methods: MDA-MB-231, MV4-11 and Raji cell lines were purchased from the American Type Culture Collection (ATCC). Conventional Western Blot was used for detection as follows. MDA-MB-231 (density 1×10 5 / mL), MV4-11 and Raji cells (density 5×10 5 / mL) were inoculated in 12-well plates. After the overnight growth was stabilized, the test compound was added for 0-24 hours, the cells were lysed to extract the protein, and boiled for 10 minutes to denature it. Take an appropriate amount of protein sample for SDS-PAGE electrophoresis. After the end, the protein in the gel was transferred to the nitrocellulose membrane using a semi-dry electrotransfer system, and the membrane was placed in a blocking solution (5% skim milk powder diluted in TBS containing 0.1% Tween 20) for 1 hour at room temperature. Then the membrane was placed in a primary antibody solution (1:1000 diluted in blocking solution) and incubated overnight at 4°C. Wash three times with TBS containing 0.1% Tween 20, each time for 5 minutes. The membrane was placed in a secondary antibody solution (horseradish peroxidase-labeled goat anti-rabbit / mouse IgG, 1:2000 diluted in blocking solution) and reacted at room temperature for 1 hour. After washing the membrane three times as above, use ECL plus reagent to develop color, and take pictures with Amersham Imager 600system.

[0622] Figure 4 The results showed that compound 2-7aa could degrade FLT3 protein in MV4-11 cells, but did not degrade the expression levels of off-target proteins AXL, ALK and LTK, and had good selectivity.

[0623] Example 88: Apoptosis-inducing effect of compound 2-7aa on acute myeloid leukemia cell line MV4-11

[0624] Methods: Cell apoptosis detection kit (BD Pharmingen TM PE Annexin V Apoptosis Detection Kit I) was purchased from BD. MV4-11 cells (1×10 6 / well) were inoculated in a six-well plate and cultured overnight at 37°C and 5% CO2. After treatment with different concentrations of compounds and positive control gilteritinib for 48 hours, the cells were collected and centrifuged. After washing with PBS, the cells were suspended in 500 μL of 1× binding buffer containing 5 μL Annexin V and 5 μL 7-AAD, reacted at room temperature in the dark for 15 minutes, filtered, and analyzed by flow cytometry (BD FACSCelesta TM ) for apoptosis detection, and protein samples treated with compounds for 48 hours were analyzed by immunoblotting.

[0625] Figure 5 The results showed that 2-7aa had a stronger ability to induce cell apoptosis than gilteritinib, and significantly affected the expression of apoptosis-related proteins, leading to the downregulation of PARP, Caspase 3 and Caspase 9 expression, and concentration-dependently upregulating the expression of their corresponding activated forms of proteins, Cleaved PARP and Cleaved Caspase 3.

[0626] Example 89: Effect of Compound 2-7aa on the Cycle Induction of Acute Myeloid Leukemia Cell Line MV4-11

[0627] Methods: Cell cycle and apoptosis detection kit (C1052) was purchased from Bio-Tech Company, China. MV4-11 cells (1×10 6 / well) were inoculated in a six-well plate and cultured overnight at 37°C and 5% CO2. The suspended cells were treated with different concentrations of the test compound and the positive control gilteritinib for 24 hours, collected by centrifugation and washed with PBS. The cells were dispersed in 70% pre-cooled ethanol and fixed for 2 hours, centrifuged and the supernatant was discarded, and 500 μL of propidium iodide staining solution was added to each tube of cell sample, and reacted at 37°C in the dark for 30 minutes. After cell filtration, flow cytometry (BD FACSCelesta TM ) for cell cycle analysis, and the ratios of cells in different phases were calculated by built-in software.

[0628] Figure 6The results showed that compound 2-7aa blocked the early DNA synthesis stage of FLT3-ITD mutated cells and inhibited the cell mitosis process; compared with gilteritinib, 2-7aa could produce a strong apoptotic effect earlier at a lower concentration.

[0629] Example 90: In vivo pharmacokinetic study of compound 2-7aa in animals

[0630] Methods: This experiment was completed by Shanghai Medicilon. SD rats were randomly divided into three groups, 3 rats in each group, and single administration of 10 mg / kg was performed orally and intraperitoneally, and 2 mg / kg was injected intravenously. Blood was collected multiple times, and the pharmacokinetic parameters of the compound plasma samples were analyzed by LC-MS / MS method.

[0631] The results in Table 4 show that: after a single oral dose of 10 mg / kg, compound 2-7aa had no exposure in rat plasma, resulting in extremely low bioavailability and ineffective absorption. When intraperitoneally injected at a dose of 10 mg / kg, the in vivo AUC value and the peak concentration C in plasma were max They are 1612.11h*ng / mL and 151.42ng / mL respectively, and the half-life of the drug is T 1 / 2 The bioavailability was 51.46% and the absorption rate was 4.44 h, indicating that intraperitoneal administration could promote the effective absorption of compound 2-7aa.

[0632] Table 4. Pharmacokinetic data of compound 2-7aa in rats

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[0634] Example 91: In vivo animal pharmacodynamics study of compound 2-7aa

[0635] Methods: This experiment was performed by Shanghai Medicilon. MV4-11 cells were cultured in IMDM medium containing 10% FBS and were stably subcultured at 37°C and 5% CO2. Cells in the logarithmic growth phase were resuspended in complete medium and Matrigel was added to adjust the cell concentration to 5×10 7 / mL, and inoculated under sterile conditions into the right back of NOD / SCID immunodeficient female mice subcutaneously at an inoculation concentration of 5×10 6 / 0.1mL / mouse. When the tumor volume reaches 200mm 3At the time of the experiment, the animals were randomly divided into a control group and an experimental group according to the tumor volume. The drug group was intraperitoneally injected with the test compound or orally administered with the positive drug gilteritinib every day according to the animal weight, and the control group was intraperitoneally injected with an equal amount of normal saline in the same way. The clinical symptoms of the animals were observed and recorded every day, and the animal weight and tumor volume were measured. After 21 days, the mice in each group were killed, and the subcutaneous tumors were photographed and weighed, and the experimental results were analyzed.

[0636] Figure 7 The results showed that: after comparison with the blank control group, it was found that compound 2-7aa could effectively inhibit the growth of MV4-11 xenograft tumors in vivo, without significant changes in the weight of mice, and had low toxicity; the positive compound gilteritinib could completely regress the tumor after administration, showing a good anti-tumor effect.

[0637] The above-mentioned embodiments only express several implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

[0638] All documents mentioned in the present invention 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 teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A pyrazinecarboxamide compound, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, a deuterated product thereof or a prodrug molecule thereof, characterized in that: The compound has the structure shown in formula (I): in: Y is selected from the group consisting of -CH2-, -C(=O)-; R1 and R2 are each independently selected from the following group: -H, -F; L has the structure shown below: -Z-(L1) m -; wherein m is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30; The Z is selected from the following group: a chemical bond, a 3-12 membered ring having 0-4 heteroatoms, -NH-, -O-, -C(O)-, -CONH-, -NHCO-, -S-, -CH2-, -C≡C-, -CH=CH-, -P=O-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-; Each L1 is independently selected from the following group: a 3-12 membered ring having 0-4 heteroatoms, -NH-, -O-, -C(O)-, -CONH-, -NHCO-, -S-, -CH2-, -C≡C-, -CH=CH-, -P=O-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-; The L segment may be optionally replaced by one or more R L Substituted; wherein said R L Selected from the following group: halogen, -OH, -CN, -CF3, -NH2, -COOH, oxygen atom (=O), substituted or unsubstituted C1-C6 alkyl or haloalkyl, substituted or unsubstituted C1-C6 alkoxy; or, 2 R L The radicals and the atoms to which they are attached together form a 3-8 membered ring containing 0-4 heteroatoms; Ring A is a substituted or unsubstituted 5-7 membered heterocyclic ring; L2 is a 4-7 membered heterocyclic ring or is absent; The substitution refers to that one or more H atoms on the group are replaced by a substituent selected from the group consisting of halogen and deuterium atom.

2. The compound according to claim 1, its pharmaceutically acceptable salt, its stereoisomer or its prodrug molecule, characterized in that: The compound of formula I has a structure selected from the following group: Wherein, the definitions of Y, L, R1 and R2 are as described in claim 1.

3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that: L has the structure shown below: in: Z is selected from the following group: a 3-8 membered ring having 0-4 heteroatoms, -NH-, -O-, -C(O)-, -CONH-, -NHCO-, -S-, -CH2-, -C≡C-, -CH=CH-, -P=O-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-; n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; And the dashed line indicates the point of attachment of the linker moiety.

4. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that: L has the structure shown below: in: W1 and W2 are absent or are each independently a 3-12 membered ring having 0-4 heteroatoms, which is optionally replaced by R L replace; R L The definition of as in claim 1; B is selected from the following group: a 3-12 membered ring that is absent or has 0-4 heteroatoms, -NH-, -O-, -CONH-, -NHCO-, -S-, -C≡C-, -CH=CH-, -P=O-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-, C1-C6 alkyl or haloalkyl (straight chain, branched, optionally substituted) and optionally one or more C atoms are replaced by O, N; or C1-C6 alkoxy (straight chain, branched, optionally substituted); Z is selected from the following group: a 3-12 membered ring containing 0-4 heteroatoms, -NH-, -O-, -CONH-, -NHCO-, -S-, -CH2-, -C≡C-, -CH=CH-, -P=O-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-; each n is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; and Dashed lines indicate points of attachment of linker moieties.

5. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that: The Z is selected from the following groups: -NHCO-, -CH2-, -C(O)-, a 3-12 membered ring containing 0-4 heteroatoms, -NH-, and -O-. In another preferred embodiment, the 3-12 membered ring having 0-4 heteroatoms is selected from the following group:

6. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, characterized in that: The L has a structure selected from the following group: wherein n and m are each independently 0, 1, 2, 3, 4, 5 or 6.

7. The compound according to claim 1, its pharmaceutically acceptable salt, its stereoisomer or its prodrug molecule, characterized in that: The compound is selected from the following group: In another preferred embodiment, the compound is selected from the following group:

8. A pharmaceutical composition for treating tumors, characterized in that The invention comprises the compound according to claim 1 or a pharmaceutically acceptable salt thereof, or a stereoisomer, a deuterated product thereof or a prodrug molecule thereof, and an optional pharmaceutically acceptable carrier.

9. Use of the compound according to claim 1, its pharmaceutically acceptable salt, its stereoisomer, its deuterated product or its prodrug molecule, characterized in that: Used for treating or preventing diseases or conditions related to the activity or expression of FLT3; wherein the diseases or conditions are selected from the following group: cardiovascular disease, diabetes, hypertension, muscular dystrophy, Parkinson's disease, Alzheimer's disease, and cancer.

10. The use according to claim 9, characterized in that The cancers include: hematological tumors, gastrointestinal stromal tumors, histiocytic lymphoma, non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, and nasopharyngeal carcinoma. In another preferred embodiment, the invention is used to treat an indication selected from the group consisting of non-small cell lung cancer, malignant melanoma, prostate cancer, kidney cancer, bladder cancer, ovarian cancer, colon cancer, rectal cancer, breast cancer, cervical cancer, lung cancer, laryngeal cancer, nasopharyngeal cancer, pancreatic cancer, multiple myeloma, B lymphoma, leukemia, or to prevent tumor recurrence after surgery.

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