2,5-diketopiperazine-imidazole compounds, methods of making and using the same

Abstract

The application discloses 2,5-diketopiperazine-imidazole compounds, a preparation method and application thereof. Specifically disclosed are compounds as shown in formula (I), a tautomer thereof, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, or a solvate of any one of the foregoing. The compound of the application is novel in structure and has good activity.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical chemistry, and particularly relates to 2,5-diketopiperazine-imidazole compounds, a preparation method thereof and application thereof. BACKGROUND

[0002] Plinabulin (NPI-2358) belongs to 2,5-diketopiperazine compound derivatives, which is a derivative of low molecular weight cyclic dipeptide Phenylahistin produced by marine fungus Aspergillus sp. through structural modification, and is a microtubule protein binding agent. Plinabulin can selectively act on the vicinity of the colchicine binding site in endothelial microtubule protein, inhibit microtubule protein polymerization, block microtubule assembly, thereby destroying the cytoskeleton of endothelial cells and inhibiting tumor blood flow. Experiments show that plinabulin acts on cells to make cells stop at the early stage of mitosis, and induces cell death. Recent studies have found that plinabulin is a differentiation immune and stem cell regulator, and a guanine nucleotide exchange factor (GEF-H1) activator, which can target and change the tumor microenvironment and destroy tumor blood vessels through multiple mechanisms; as an effective antigen-presenting cell (APC) inducer (by activating dendritic cell maturation), its persistent anticancer effect is related to its T cell activation effect (Cell Reports 2019, 28:13, 3367-3386).

[0003] At present, the candidate drug is developed by BeyondSpring Pharmaceutical Company, and is undergoing clinical phase III experiment, which is used for treating non-small cell lung cancer in combination with docetaxel, and is used for preventing chemotherapy-induced neutropenia (CIN) in non-myeloid malignancies. The indication of whitening has also been applied for NDA.

[0004] The chemical structural formula of plinabulin is as follows:

[0005]

[0006] The molecular formula of plinabulin is C 19 H 20 N4O2, the molecular weight is 336.39, the CAS number is 714272-27-2. It has good stability, but poor water solubility, large toxic side effects, and still needs to continue to develop high-activity microtubule inhibitors. SUMMARY

[0007] The technical problem to be solved by the present application is the defects of plinabulin, and 2,5-diketopiperazine-imidazole compounds, a preparation method thereof and application thereof are provided. The compound of the present application has novel structure and good activity.

[0008] To achieve the above object, the present application adopts the following technical solutions:

[0009] The present application provides a compound as shown in formula (I), a tautomer thereof, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, or a solvate of any of the foregoing (referring to the compound as shown in formula (I), the tautomer thereof, the stereoisomer thereof or the pharmaceutically acceptable salt thereof):

[0010]

[0011] wherein,

[0012] R 1 is hydrogen, deuterium, halogen, C1-C8alkyl, C1-C8alkyl substituted by one or more halogens, C1-C8alkoxy, C1-C8alkoxy substituted by one or more halogens, C2-C8alkenyl, benzoyl, benzoyl substituted by one or more halogens, phenoxy or "phenoxy substituted by one or more halogens";

[0013] R 1a , R 1b and R 1c are independently hydrogen, deuterium, halogen or C1-C8alkyl;

[0014] Alternatively, R 1 , R 1a together with the carbon atom to which they are attached form a C6-C 10 aryl group or a "3-10 membered heteroaryl group having 1-5 heteroatoms selected from N, O and S";

[0015] Alternatively, R 1 , R 1b together with the carbon atom to which they are attached form a C6-C 10 aryl group or a "3-10 membered heteroaryl group having 1-5 heteroatoms selected from N, O and S";

[0016] Alternatively, R 1b , R 1c together with the carbon atom to which they are attached form a C6-C 10 aryl group or a "3-10 membered heteroaryl group having 1-5 heteroatoms selected from N, O and S";

[0017] R 2 is hydrogen, deuterium, C1-C8alkyl substituted by one or more halogens, C1-C8alkyl or C3-C 10 cycloalkyl;

[0018] L is "Ci-C8-heteroalkylene with one or more heteroatoms selected from the group consisting of N, O, S and Se in a number ranging from 1 to 4, the number of heteroatoms being 1 to 4", Ci-C8-alkylene, or "Ci-C8-alkylene substituted by one or more R 2-2 substituted "Ci-C8-heteroalkylene with one or more heteroatoms selected from the group consisting of N, O, S and Se in a number ranging from 1 to 4, the number of heteroatoms being 1 to 4", Ci-C8-alkylene, or "Ci-C8-alkylene substituted by one or more R 2-1 substituted Ci-C8-alkylene" ;

[0019] R 2-1 and R 2-2 are independently deuterium, OH, Ci-C8-alkoxy, or "Ci-C8-alkoxy substituted by one or more R 2-1-1 substituted Ci-C8-alkoxy" ;

[0020] R 2-1-1 are independently C3-C 10 cycloalkyl, C6-C 10 aryl, or "3-10 membered heteroaryl with one or more heteroatoms selected from the group consisting of N, O and S in a number ranging from 1 to 5, the number of heteroatoms being 1 to 5" ;

[0021] Ring A is C6-C 10 aryl, "3-10 membered heteroaryl with one or more heteroatoms selected from the group consisting of N, O and S in a number ranging from 1 to 5, the number of heteroatoms being 1 to 5", "3-10 membered heteroaryl with one or more heteroatoms selected from the group consisting of N, O and S in a number ranging from 1 to 5, the number of heteroatoms being 1 to 5, substituted by one or more R A-1 substituted C6-C 10 aryl, "3-10 membered heteroaryl with one or more heteroatoms selected from the group consisting of N, O and S in a number ranging from 1 to 5, the number of heteroatoms being 1 to 5", "3-10 membered heteroaryl with one or more heteroatoms selected from the group consisting of N, O and S in a number ranging from 1 to 5, the number of heteroatoms being 1 to 5, substituted by one or more R A-2 substituted 3-10 membered heteroaryl with one or more heteroatoms selected from the group consisting of N, O and S in a number ranging from 1 to 5, the number of heteroatoms being 1 to 5", C3-C 10 cycloalkyl, "3-10 membered heterocycloalkyl with one or more heteroatoms selected from the group consisting of N, O and S in a number ranging from 1 to 5, the number of heteroatoms being 1 to 5", or "3-10 membered heterocycloalkyl with one or more heteroatoms selected from the group consisting of N, O and S in a number ranging from 1 to 5, the number of heteroatoms being 1 to 5, substituted by one or more R A-3 substituted C3-C 10 cycloalkyl, "3-10 membered heterocycloalkyl with one or more heteroatoms selected from the group consisting of N, O and S in a number ranging from 1 to 5, the number of heteroatoms being 1 to 5", or "3-10 membered heterocycloalkyl with one or more heteroatoms selected from the group consisting of N, O and S in a number ranging from 1 to 5, the number of heteroatoms being 1 to 5, substituted by one or more R A-4 substituted 3-10 membered heterocycloalkyl with one or more heteroatoms selected from the group consisting of N, O and S in a number ranging from 1 to 5, the number of heteroatoms being 1 to 5" ;

[0022] R A-1 , R A-2 , R A-3 and R A-4 are independently deuterium, halogen, hydroxyl, cyano, nitro, Ci-C8-alkyl, Ci-C8-alkyl substituted by one or more halogens, Ci-C8-alkoxy, Ci-C8-alkoxy substituted by one or more halogens, C3-C 10 cycloalkyl, C6-C 10 aryl, "3-10 membered heteroaryl with one or more heteroatoms selected from the group consisting of N, O and S in a number ranging from 1 to 5, the number of heteroatoms being 1 to 5", "3-10 membered heteroaryl with one or more heteroatoms selected from the group consisting of N, O and S in a number ranging from 1 to 5, the number of heteroatoms being 1 to 5, substituted by one or more R A-1-1 substituted C6-C 10 aryl, or -C(=O)-R3-1-2 ;

[0023] R 3-1-2 is C1-C8alkyl, C1-C8alkyl substituted by one or more halogen, C6-C10aryl, C6-C10aryl substituted by one or more R 10 ; 3-1-2-1 ; 10 ;

[0024] R A-1-1 and R 3-1-2-1 are independently halogen, hydroxy, cyano, nitro, C1-C8alkyl, C1-C8alkyl substituted by one or more halogen, C1-C8alkoxy, C1-C8alkoxy substituted by one or more halogen or C3-C7cycloalkyl. 10 ;

[0025] In a preferred embodiment of the present application, certain groups in the compounds of formula (I), the tautomers thereof, the stereoisomers thereof, the pharmaceutically acceptable salts thereof or the solvates of any of the foregoing (simply referred to as "in a preferred embodiment of the present application"), are defined as follows, and the groups not mentioned are as described in any of the embodiments of the present application (simply referred to as "in an embodiment of the present application"),

[0026] when R 1 is halogen, said halogen is fluorine.

[0027] In an embodiment of the present application, when R 1 is C1-C8alkyl, said C1-C8alkyl is C1-C4alkyl, for example methyl.

[0028] In an embodiment of the present application, when R 1 is C1-C8alkyl substituted by one or more halogen, said C1-C8alkyl substituted by one or more halogen is C1-C4alkyl substituted by one or more halogen, for example trifluoromethyl.

[0029] In an embodiment of the present application, when R 1 is C1-C8alkoxy, said C1-C8alkoxy is C1-C4alkoxy, for example methoxy.

[0030] In an embodiment of the present application, when R 1 is C1-C8alkoxy substituted by one or more halogen, said C1-C8alkoxy substituted by one or more halogen is C1-C4alkoxy substituted by one or more halogen, for example trifluoromethoxy.

[0031] In an embodiment of the present application, when R 1When R is phenyl substituted by one or more halogen, the halogen is fluorine. Preferably, the phenyl substituted by one or more halogen is

[0032] In a certain embodiment of the application, when R 1 is phenyl substituted by one or more halogen, the halogen is fluorine. Preferably, the phenyl substituted by one or more halogen is

[0033] In a certain embodiment of the application, when R 1a , R 1b and R 1c are independently halogen, the halogen is fluorine.

[0034] In a certain embodiment of the application, when R 1a , R 1b and R 1c are independently C1-C8 alkyl, the C1-C8 alkyl is C1-C4 alkyl, e.g. methyl.

[0035] In a certain embodiment of the application, when R 1 , R 1a together with the carbon atom to which they are attached form C6-C 10 aryl, the C6-C 10 aryl is phenyl.

[0036] In a certain embodiment of the application, when R 1 , R 1a together with the carbon atom to which they are attached form "3-10 membered heteroaryl having 1-5 heteroatoms selected from the group consisting of N, O and S", the "3-10 membered heteroaryl having 1-5 heteroatoms selected from the group consisting of N, O and S" is "5-6 membered heteroaryl having 1-2 heteroatoms selected from the group consisting of N, O and S"; e.g. pyridyl.

[0037] In a certain embodiment of the application, when R 2 is C1-C8 alkyl, the C1-C8 alkyl is C1-C4 alkyl, e.g. methyl, ethyl, isopropyl or tert-butyl.

[0038] In a certain embodiment of the application, when R 2 is C3-C 10 cycloalkyl, the C3-C 10 cycloalkyl is C3-C6 cycloalkyl, e.g. cyclopropyl.

[0039] In one aspect of the present invention, when L is "a C1-C8 heteroalkylene group whose heteroatoms are selected from one or more of N, O, S, and Se, and whose heteroatoms number 1-4", the phrase "a C1-C8 heteroalkylene group whose heteroatoms are selected from one or more of N, O, S, and Se, and whose heteroatoms number 1-4" is interpreted as "a C1-C5 heteroalkylene group whose heteroatoms are selected from one or more of O and S, and whose heteroatoms number 1-2". For example...

[0040] In one aspect of the present invention, when L is a C1-C8 alkylene group, the C1-C8 alkylene group is a C1-C5 alkylene group, such as methylene.

[0041] In one aspect of this invention, when ring A is C6-C 10 aryl or "by one or more R A-1 Replacement C6-C 10 When "aryl", the C6-C 10 The aryl group is phenyl or naphthyl (e.g. ), for example, phenyl.

[0042] In one aspect of the present invention, ring A is "a 3-10 membered heteroaryl group whose heteroatoms are selected from one or more of N, O and S, and whose number of heteroatoms is 1-5" or "is surrounded by one or more R..." A-2 When the substituted heteroatom is selected from one or more of N, O, and S, and the number of heteroatoms is 1-5 in a 3-10 membered heteroaryl group, the phrase "the heteroatom is selected from one or more of N, O, and S, and the number of heteroatoms is 1-5 in a 3-10 membered heteroaryl group" is translated as "the heteroatom is selected from one or more of N, O, and S, and the number of heteroatoms is 1-2 in a 5-10 membered heteroaryl group," for example, pyridyl, indolyl, indololinyl, pyrroleyl, or tetrahydroquinolinyl, and for another example...

[0043] In one aspect of this invention, when ring A is C3-C 10 cycloalkyl or "by one or more R A-3 Replacement C3-C 10 When "cycloalkyl", the C3-C 10 The cycloalkyl group is a C3-C6 cycloalkyl group, such as a cyclopentyl group.

[0044] In one aspect of the present invention, ring A is "a 3-10 membered heterocyclic alkyl group with heteroatoms selected from one or more of N, O and S, and having 1-5 heteroatoms" or "is surrounded by one or more R..." A-4When the substituted heteroatom is selected from one or more of N, O, and S, and the heteroatom number is 1-5, and it is a 3-10 membered heterocyclic alkyl group, then the "heteroatom selected from one or more of N, O, and S, and the heteroatom number is 1-5," is defined as "the heteroatom selected from one or more of N, O, and S, and the heteroatom number is 1-2, and it is a 5-6 membered heterocyclic alkyl group," for example, pyrrolidinyl or piperidinyl, and for another example...

[0045] In one aspect of the present invention, when R A-1 R A-2 R A-3 and R A-4 When it is a halogen on its own, the halogen is fluorine, chlorine, or bromine.

[0046] In one aspect of the present invention, when R A-1 R A-2 R A-3 and R A-4 When independently a C1-C8 alkyl group, the C1-C8 alkyl group is a C1-C4 alkyl group, such as methyl.

[0047] In one aspect of the present invention, when R A-1 R A-2 R A-3 and R A-4 When independently a C1-C8 alkyl group substituted with one or more halogens, the C1-C8 alkyl group substituted with one or more halogens is a C1-C4 alkyl group substituted with one or more halogens, such as trifluoromethyl.

[0048] In one aspect of the present invention, when R A-1 R A-2 R A-3 and R A-4 When independently a C1-C8 alkoxy group, the C1-C8 alkoxy group is a C1-C4 alkoxy group, such as a methoxy group.

[0049] In one aspect of the present invention, when R A-1 R A-2 R A-3 and R A-4 When independently a C1-C8 alkoxy group is substituted with one or more halogens, the C1-C8 alkoxy group substituted with one or more halogens is a C1-C4 alkoxy group substituted with one or more halogens, such as a trifluoromethoxy group.

[0050] In one aspect of the present invention, when R A-1 R A-2 R A-3 and R A-4 Independently for C6-C 10C6-C10aryl or "C6-C10aryl substituted by one or more R A-1-1 C6-C10aryl or "C6-C10aryl substituted by one or more R 10 C6-C10aryl or "C6-C10aryl substituted by one or more R 10 C6-C10aryl is phenyl.

[0051] In a certain embodiment of the present application, R 3-1-2 C1-C8alkyl or C3-C8cycloalkyl, preferably C1-C8alkyl or C3-C8cycloalkyl substituted by one or more halogen.

[0052] In a certain embodiment of the present application, R 3-1-2 C6-C10aryl or "C6-C10aryl substituted by one or more R 10 C6-C10aryl or "C6-C10aryl substituted by one or more R 3-1-2-1 C6-C10aryl or "C6-C10aryl substituted by one or more R 10 C6-C10aryl or "C6-C10aryl substituted by one or more R 10 C6-C10aryl is phenyl.

[0053] In a certain embodiment of the present application, R 1 halogen, benzoyl substituted by one or more halogen or "phenoxy substituted by one or more halogen", for example R 1 halogen, benzoyl substituted by one or more halogen or "phenoxy substituted by one or more halogen", for example R

[0054] In a certain embodiment of the present application, R 1a halogen.

[0055] In a certain embodiment of the present application, R 1b halogen.

[0056] In a certain embodiment of the present application, R 1c halogen.

[0057] In a certain embodiment of the present application, R 2 C1-C8alkyl or C3-C8cycloalkyl, preferably C1-C8alkyl or C3-C8cycloalkyl substituted by one or more halogen. 10 C1-C8alkyl or C3-C8cycloalkyl, preferably C1-C8alkyl or C3-C8cycloalkyl substituted by one or more halogen. 10 C1-C8alkyl or C3-C8cycloalkyl, preferably C1-C8alkyl or C3-C8cycloalkyl substituted by one or more halogen.

[0058] In a certain embodiment of the present application, L is "C1-C8heteroalkylene with one or more heteroatoms selected from the group consisting of N, O, S and Se in the number ranging from 1 to 4" or C1-C8alkylene, preferably C1-C8alkylene.

[0059] In a certain embodiment of the present application, ring A is C6-C10aryl, C6-C10aryl substituted by one or more R 10 C6-C10aryl, C6-C10aryl substituted by one or more R A-1 C6-C10aryl, C6-C10aryl substituted by one or more R 10 3-10 membered heteroaryl with one or more heteroatoms selected from the group consisting of N, O and S in the number ranging from 1 to 5, C3-C8cycloalkyl, C3-C8cycloalkyl substituted by one or more R10 Cycloalkyl or "a 3-10 membered heterocycloalkyl group with 1-5 heteroatoms selected from one or more of N, O and S"; preferably C6-C 10 aryl, with one or more R A-1 Replacement C6-C 10 Aryl or "a 5-6 membered heteroaryl group selected from one or more of N, O and S, with 1-2 heteroatoms".

[0060] In one aspect of the present invention, R A-1 Independently halogen, cyano, nitro, C1-C8 alkyl, C1-C8 alkyl substituted with one or more halogens, C1-C8 alkoxy, C1-C8 alkoxy substituted with one or more halogens, C6-C 10 Aryl or -C(=O)-R 3-1-2 Preferably, it is a halogen, cyano, nitro, C1-C8 alkyl, C1-C8 alkyl substituted with one or more halogens, C1-C8 alkoxy, C1-C8 alkoxy substituted with one or more halogens, or -C(=O)-R 3-1-2 .

[0061] In one aspect of the present invention, R 3-1-2 It is a C1-C8 alkoxy or C6-C 10 Aryl; preferably C1-C8 alkoxy.

[0062] In one aspect of the present invention, for

[0063] In one embodiment of the present invention, ring A is...

[0064] In one aspect of the present invention, the compound represented by formula (I) is not any of the following compounds:

[0065]

[0066]

[0067]

[0068]

[0069]

[0070] Preferably, the compound represented by formula (I) is any of the following compounds:

[0071]

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078] The present application also provides a preparation method of the above-mentioned compound as shown in formula (I), which comprises the following steps: condensation reaction of a compound as shown in formula (II) with a compound as shown in formula (III) as shown below, to obtain the compound as shown in formula (I);

[0079]

[0080] wherein, L, R 1 , R 1a , R 1b , R 1c , R 2 and the definition of ring A is described in any of the preceding.

[0081] The present application also provides a compound as shown in formula (II):

[0082]

[0083] wherein, L, R 2 and the definition of ring A is described in any of the preceding.

[0084] In a certain scheme of the present application, the compound as shown in formula (II) is not any of the following compounds:

[0085]

[0086]

[0087]

[0088]

[0089] Preferably, the compound as shown in formula (II) is any of the following compounds:

[0090]

[0091]

[0092]

[0093] The present application also provides a method for preparing the above-mentioned compound represented by formula (II), which comprises the following step: subjecting a 2,5-diketopiperazine derivative represented by formula (A) and a compound represented by formula (B) to a substitution reaction as shown below to obtain the compound represented by formula (II);

[0094]

[0095] wherein Y is halogen or -S(=O)2-R 8 , R 8 is C1-C6 alkyl or C6-C 10 aryl; L, R 2 and ring A are as defined in any one of the preceding embodiments.

[0096] Preferably, the substitution reaction is carried out in an aprotic solvent, which can be conventional in the art, and preferably the aprotic solvent is one or more of tetrahydrofuran, dichloromethane, cyclohexane, acetonitrile, acetone, N,N-dimethylformamide, N,N-dimethylacetamide, dimethylsulfoxide, hexamethylphosphoramide, benzene, toluene, nitrobenzene, xylene, carbon tetrachloride and carbon disulfide; for example N,N-dimethylformamide.

[0097] Preferably, the substitution reaction is carried out in the presence of a base, which can be a base commonly used in such reactions in the art, and preferably the base is one or more of sodium hydride, potassium hydroxide, potassium carbonate, cesium carbonate, sodium bicarbonate, potassium bicarbonate, 1,8-diazabicycloundec-7-ene, 1,5-diazabicyclo[4.3.0]non-5-ene, triethylamine and diisopropylethylamine; for example cesium carbonate.

[0098] Preferably, the substitution reaction is carried out in the presence of a catalyst, which can be a catalyst commonly used in such reactions in the art, and preferably the catalyst is one or more of potassium iodide and sodium iodide; for example potassium iodide.

[0099] The present application also provides a pharmaceutical composition comprising the above-mentioned compound represented by formula (I), a tautomer thereof, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, or a solvate of any of the foregoing, and a pharmaceutical adjuvant.

[0100] In certain embodiments, the pharmaceutical adjuvant does not comprise a cosolvent.

[0101] The present application also provides the use of the above-mentioned compound of formula (I), a tautomer thereof, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, or a solvate of any of the foregoing, or the above-mentioned pharmaceutical composition in the manufacture of a medicament. Preferably, the medicament is for the prevention and / or treatment of cancer. The cancer is preferably one or more of lung cancer, pancreatic cancer, colon cancer and liver cancer. The compounds of the present application exhibit strong cytotoxicity against a variety of tumor cells, for example, all of the compounds of the present application exhibit proliferation inhibitory activity against NCI-H460, BXPC-3, HT-29, etc. cells with IC 50 values < 5 μM.

[0102] The present application also provides the use of the above-mentioned compound of formula (I), a tautomer thereof, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, or a solvate of any of the foregoing, or the above-mentioned pharmaceutical composition in the manufacture of a microtubulin inhibitor.

[0103] In the use, the microtubulin inhibitor can be used in vivo in a mammalian organism; it can also be used in vitro, mainly as a standard or control sample for comparison or in a kit according to the conventional methods in the art, for rapid detection of the effect of the microtubulin inhibitor.

[0104] The present application also provides a method for preventing and / or treating cancer, comprising administering to a patient a therapeutically effective amount of the above-mentioned compound of formula (I), a tautomer thereof, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, or a solvate of any of the foregoing, or the above-mentioned pharmaceutical composition.

[0105] The compounds of the present application, tautomers thereof, stereoisomers thereof, pharmaceutically acceptable salts thereof, solvates of any of the foregoing, pharmaceutical compositions can be administered topically or systemically, for example, for enteral administration, such as rectal or oral administration, or for parenteral administration to a mammal, especially a human. Exemplary compositions for rectal administration include suppositories, which can be prepared from a mixture of the present compounds and a suitable non-irritating excipient, such as a conventional solid dosage form, like cocoa butter, synthetic glyceride esters, or polyethylene glycol, which are solid at ordinary temperatures but liquefy and / or dissolve in the rectal cavity to release the drug.

[0106] A therapeutically effective amount of the active ingredient is defined in context and depends on the species, body weight, age, individual condition, individual pharmacokinetic parameters of the mammal, the disease to be treated, and the mode of administration. For enteral administration, such as oral administration, the compounds of the present application can be formulated into a wide variety of dosage forms.

[0107] The effective amount of the compound, the tautomer thereof, the stereoisomer thereof, the pharmaceutically acceptable salt thereof, the solvate of any one of the foregoing, or the pharmaceutical composition of the present application can be easily determined by routine experiments, and the most effective and convenient administration route and the most appropriate formulation can also be determined by routine experiments.

[0108] The pharmaceutical excipient can be those widely used in the pharmaceutical production field. The excipient is mainly used to provide a safe, stable and functional pharmaceutical composition, and can also provide a method for allowing the active ingredient to be dissolved at a desired rate after the subject receives administration, or to facilitate the active ingredient to be effectively absorbed after the subject receives administration of the composition. The pharmaceutical excipient can be an inert filler, or provide a certain function, such as stabilizing the overall pH of the composition or preventing the degradation of the active ingredient of the composition. The pharmaceutical excipient can include one or more of the following excipients: binders, suspending agents, emulsifying agents, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adherents, glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavorants, and sweeteners.

[0109] The substances that can be used as the pharmaceutically acceptable excipient include, but are not limited to, ion exchangers, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylate, waxes, polyethylene-polyoxypropylene-block polymers, wool fat, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, phosphate buffer solutions, and other non-toxic, suitable lubricants such as sodium lauryl sulfate and magnesium stearate, coloring agents, releasing agents, coating agents, sweetening agents, flavoring agents and perfuming agents, preservatives and antioxidants.

[0110] The pharmaceutical composition of the present application can be prepared according to the disclosure using any method known to those skilled in the art. For example, conventional mixing, dissolving, granulating, emulsifying, dragee-making, encapsulating, entrapping or lyophilizing processes.

[0111] Pharmaceutical dosage forms of the compounds of the present application can be provided in immediate release, controlled release, sustained release or targeted drug release systems. For example, common dosage forms include solutions and suspensions, (micro)emulsions, ointments, gels and patches, liposomes, tablets, dragees, soft or hard shell capsules, suppositories, ovules, implants, amorphous or crystalline powders, aerosols and lyophilized formulations. Depending on the route of administration used, special devices can be required to apply or administer the drug, such as syringes and needles, inhalers, pumps, injection pens, applicators or special flasks. Pharmaceutical dosage forms often consist of the drug, excipients and a container / sealing system. One or more excipients (also known as inactive ingredients) can be added to the compounds of the present application to improve or facilitate the manufacture, stability, administration and safety of the drug, and can provide a means to obtain the desired drug release profile. The type of excipient added to the drug can therefore depend on various factors, such as the physical and chemical properties of the drug, the route of administration and the preparation steps. Pharmaceutical excipients exist in the art and include those listed in various pharmacopeias. (See U.S. Pharmacopeia (USP), Japanese Pharmacopoeia (JP), European Pharmacopoeia (EP) and British Pharmacopoeia (BP); U.S. Food and Drug Administration (www.fda.gov) Center for Drug Evaluation and Research (CEDR) publications, such as the Inactive Ingredient Guide (1996); Handbook of Pharmaceutical Additives, by Ash and Ash (2002, Synapse Information Resources, Inc., Endicott NY; etc.).

[0112] Pharmaceutical dosage forms of the compounds of the present application can be manufactured by any of the methods well-known in the art, for example by conventional mixing, sieving, dissolving, melting, granulating, dragee-making, tabletting, suspending, extruding, spray-drying, sifting, emulsifying, (nano / micro)encapsulating, pan-coating, or lyophilizing processes. As described above, the compositions of the present application can include one or more than one physiologically acceptable inactive ingredient that facilitates the processing of the active molecules into formulations for pharmaceutical use.

[0113] The pharmaceutical compositions and dosage forms can contain one or more compounds of the present application, one or more pharmaceutically acceptable salts thereof, or one or more solvates of any of the foregoing, as an active ingredient. Pharmaceutically acceptable carriers can be either solid or liquid. Solid forms include powders, tablets, pills, capsules, caplets, cachets, suppositories, and dispersible granules. A solid carrier can also serve as a diluent, flavoring agent, solubilizer, lubricant, suspending agent, binder, preservative, tablet disintegrant, or an encapsulating material. In powders, the carrier is a finely divided solid, which is in a mixture with the finely divided active ingredient. In tablets, the active ingredient is mixed with the carrier having the necessary binding capacity in suitable proportions and compacted in the shape and size desired. Suitable carriers include, but are not limited to, magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, methylcellulose, sodium carboxymethylcellulose, low-melting wax, cocoa butter, and the like. Formulations of active compounds can include encapsulating materials as carriers, providing a capsule in which active ingredient, with or without carriers, is enclosed.

[0114] Other forms adapted to oral administration include liquid form preparations including emulsions, syrups, elixirs, aqueous solutions, aqueous suspensions, or solid form preparations which are intended to be converted shortly before use to liquid form preparations. Emulsions can be prepared in solutions, for example, in aqueous propylene glycol solutions or can contain emulsifying agents, such as lecithin, sorbitan monoleate, or acacia. Aqueous solutions can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizing, and thickening agents. Aqueous suspensions can be prepared by dispersing the finely divided active component in water with a surfactant to aid suspension. Solid form preparations include solutions, suspensions, and emulsions, and can contain colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, and the like.

[0115] For parenteral administration, the pharmaceutical compositions of the present application can be in the form of sterile injectable or infusible solutions or suspensions, for example, as a sterile aqueous or oleaginous suspension. This suspension can be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable or infusible preparations can also be sterile injectable or infusible solutions or suspensions in a non-toxic parenterally acceptable diluent or solvent. For example, the pharmaceutical compositions can be solutions in 1,3-propanediol. Other examples of acceptable vehicles and solvents that can be used in the pharmaceutical compositions of the present application include, but are not limited to, mannitol, water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions can also contain a long-chain alcohol diluent or dispersant. Solutions for parenteral use can also include suitable stabilizing agents, and if desired, buffer substances. Suitable stabilizing agents include antioxidants such as, individually or in combination, sodium bisulfite, sodium sulfite, or ascorbic acid, citric acid and its salts, and EDTA sodium salt. Parenteral solutions can also contain preservatives, such as benzalkonium chloride, parabens, and chlorobutanol.

[0116] Therapeutically effective doses can be estimated initially using a variety of methods well-known in the art. Initial dosages for use in animals can be based on effective concentrations found to be

[0117] An effective amount or therapeutically effective amount or dose of an agent, such as a compound of the present application, refers to an amount of the agent or compound that will elicit the improvement of symptoms or prolongation of survival in an individual. Toxicity and therapeutic efficacy of such molecules can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., by measuring the LD 50 (the dose that is lethal to 50% of the population) and the ED 50 (the dose that is therapeutically effective for 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD 50 / ED 50 Agents that exhibit large therapeutic indices are preferred.

[0118] An effective amount or a therapeutically effective amount is the amount of a compound or pharmaceutical composition that will elicit the biological or medical response of a tissue, system, animal, or human that is being sought by a researcher, veterinarian, medical doctor, or other clinician. The dose is preferably within a range of circulating concentrations that include the ED 50 of minimal toxicity or no toxicity. The dosage can vary within this range depending upon the dosage form employed and / or the route of administration utilized. The proper dosage and frequency for any particular case depends on the particular condition being treated, as well as on the individual patient's characteristics such as general health, age, sex, diet, and weight. The particular dosage for any given case will depend upon such factors as the severity and type of disease, the patient's physical status, the particular compound or pharmaceutical composition used, and the mode of administration.

[0119] The dose and interval between doses can be adjusted individually to provide plasma levels of the active moiety that are sufficient to achieve the desired effects; i.e., the minimal effective concentration (MEC). The MEC will vary for each compound but can be estimated from in vitro data and from animal experiments. The dosages necessary to achieve the MEC will vary depending on individual characteristics and route of administration. In the case of local administration or selective uptake, the effective local concentration of the drug can not be related to plasma concentration.

[0120] The amount of the agent or composition administered can vary depending on factors such as the gender, age, and weight of the individual being treated, the severity of the affliction, the manner of administration, and the judgment of the prescribing physician.

[0121] As used herein, the terms have the following meanings unless otherwise indicated:

[0122] As will be understood by those skilled in the art, the use of in the structural formulae of the groups described herein means that the corresponding group is attached to the rest of the molecule by this point.

[0123] As used herein, the terms can be preceded and / or followed by a single dash "-", or a double dash " = " indicating the order of the bond between the named substituent and the parent moiety; a single dash indicates a single bond, and a double dash indicates a double bond. In the absence of a single or double dash, a single bond between the substituent and the parent moiety is to be assumed; furthermore, the substituents are to be read "from left to right" unless otherwise indicated.

[0124] The term "plurality" means 2, 3, 4, or 5, preferably 2 or 3.

[0125] The term "pharmaceutically acceptable" means that the salt, solvent, adjuvant, etc. is not toxic or safe and appropriate for use in patients. The "patients" are preferably mammals, more preferably humans.

[0126] The term "solvate" means a physical association or a bond between a compound of this application and one or more solvent molecules, which can be or can not be water. This physical association can, for example, be due to hydrogen bonding, to van der Waals forces, to ionic interactions, or to a combination thereof. Solvates are generally identifiable by their X-ray crystal structures, by their melting point, or by their spectroscopic properties, such as IR, NMR, and Raman spectra. The term "solvate" includes, but is not limited to, sulfates, methanolic, ethanolic, and the like.

[0127] The terms "pharmaceutically acceptable salt" and "solvate" of a pharmaceutically acceptable salt" in the term "solvate of a pharmaceutically acceptable salt" mean, as described above, a compound of this application prepared with 1, with a relatively nontoxic, pharmaceutically acceptable acid or base, or 2, with a stoichiometric or non-stoichiometric amount of a solvent.

[0128] When any variable (e.g., R 4-1 ) occurs more than one time in a compound, its definition in each occurrence is independent of its meaning in every other instance. Thus, if a group is substituted with 1, 2, or 3 R 4-1 groups, that is to say, the group can be substituted with up to 3 R 4-1 groups, the definition of R 4-1 at each occurrence is independent of the definition at every other occurrence. 4-1 In addition, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0129] The term "halogen" means fluorine, chlorine, bromine, or iodine.

[0130] The term "alkyl" means a straight or branched chain alkyl group having the number of carbon atoms indicated. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like.

[0131] The term "alkylene" means a divalent group derived from the formal removal of two monovalent or one divalent atom or group from a straight or branched chain alkane. The two valences can be on the same carbon atom or on different carbon atoms (e.g., the two valences are on the two terminal carbon atoms). For example, methylene can be (-CH2-), and ethylene can be -CH2CH2- or -CH(CH3)-.

[0132] The term "heteroalkylene" means a divalent group derived from the formal removal of two monovalent or one divalent atom or group from a straight or branched chain heteroalkane. The two valences can be on the same atom or on different atoms (e.g., the two valences are on the two terminal atoms). For example, propylene containing an oxygen atom can be and the like.

[0133] The term "alkoxy" means the group -O-R X where R Xalkyl as defined above.

[0134] The term "alkenyl" refers to straight-chain or branched hydrocarbon groups having one or more carbon-carbon double bonds and no carbon-carbon triple bonds, the one or more carbon-carbon double bonds can be internal or terminal, examples of alkenyl groups include ethenyl, allyl, methyl-ethenyl, propenyl, butenyl, pentenyl, 1,1-dimethyl-2-propenyl, hexenyl, and the like.

[0135] The term "aryl" refers to a C6-Ci2carbocyclic ring having a completely conjugated pi- electron system. Examples of aryl groups include phenyl and naphthyl. 10 aryl, such as phenyl or naphthyl.

[0136] The term "heteroaryl" refers to an aromatic group containing heteroatoms, preferably 1-5 aromatic 3-6 membered monocyclic or 9-10 membered bicyclic rings, at least one ring of which has aromaticity, when bicyclic, for example, furanyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thiophenyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, thiadiazolyl, benzimidazolyl, indolyl, indazolyl, indolinyl, benzothiazolyl, benzoisothiazolyl, benzoxazolyl, benzoisoxazolyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, and the like.

[0137] The term "cycloalkyl" refers to a saturated cyclic alkyl group, for example, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0138] The term "heterocycloalkyl" refers to a saturated cyclic group having heteroatoms, preferably 1-5 ring heteroatoms independently selected from N, O, and S, in a 3-10 membered saturated monocyclic or bicyclic ring. Examples of heterocycloalkyl groups are: tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, tetrahydropyridinyl, tetrahydropyrrolyl, azetidinyl, thiazolidinyl, oxazolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, azepanyl, diazepanyl, oxazepanyl, and the like. Preferred heterocyclyl groups are morpholin-4-yl, piperidin-1-yl, pyrrolidin-1-yl, thiomorpholin-4-yl, and 1,1-dioxo-thiomorpholin-4-yl.

[0139] The term "treatment" refers to therapeutic treatment. When used in relation to a particular condition, treatment refers to: (1) alleviating one or more of the signs or symptoms of the disease or condition, (2) interfering with (a) one or more of the points in a biological cascade that leads to or causes the condition or (b) one or more of the signs of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or with treatment of the condition, or (4) slowing the development of the condition or one or more of the signs of the condition.

[0140] The term "prevention" refers to a reduction in the risk of acquiring or developing a disease or disorder.

[0141] The term "patient" refers to any animal, preferably a mammal, and most preferably a human, to which a compound or composition according to embodiments of the present application is to be administered or has been administered. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, and the like, with humans being most preferred.

[0142] The above-mentioned preferred conditions can be combined in any way, without departing from the common general knowledge of the skilled person, to give further preferred embodiments of the present application.

[0143] The reagents and starting materials used in the present application are commercially available.

[0144] The positive progress effect of the present application is that:

[0145] The compounds involved in the present application are novel compounds obtained by the inventors of the present application through structural and synthetic route design, chemical synthesis, and have not been reported in the literature. Compared with the control Plinabulin, some of the compounds have anti-tumor activity comparable to or even better than Plinabulin, and have good development prospects. BRIEF DESCRIPTION OF DRAWINGS

[0146] Figure 1 Immunofluorescence results of 2,5-diketopiperazine-imidazole compound H460 cell lines. DETAILED DESCRIPTION

[0147] The present application will be further described below by way of examples, but the present application is not limited to the scope of the examples described.

[0148] Example 1 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-(5-tert-butyl-1- benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-1)

[0149]

[0150] 1) Take 50 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-tert-butyl-1H-imidazol-4- yl)methylene)piperazine-2,5-dione (500 mg, 1.72 mmol), potassium carbonate (714.04 mg, 5.17 mmol), DMF (8 ml) in turn, stir at room temperature, then add benzyl bromide (589.13 mg, 3.62 mmol), continue to stir at 60 °C for 2.5 h. The reaction solution is dropped into 4 °C cold water, a solid is precipitated, suction filtration, filter cake water washing, 50 °C vacuum drying, methanol ultrasonic slurry, placed in -30 °C refrigerator overnight, suction filtration, filter cake with cold methanol washing, 50 °C vacuum drying, white solid 185 mg, yield 28.24%.

[0151] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-tert-butyl-1-benzylimidazol-4- yl)methylene)piperazine-2,5-dione (150 mg, 0.40 mmol), 3-p-fluorobenzoyl benzaldehyde (107.97 mg, 0.47 mmol), cesium carbonate (192.69 mg, 0.59 mmol), anhydrous sodium sulfate (112.13 mg, 0.79 mmol), DMF (3 ml) in turn, exhaust, nitrogen protection, placed in 45 °C oil bath, stir for 20 h. LC-MS monitoring reaction, raw material point disappears. After the reaction, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtration, filter cake cold water washing (30 ml*2), the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure. The water phase is extracted with EA until clear, the organic phases are combined and concentrated under reduced pressure. The filter cake and organic phase sample are combined, ultrasonic slurry with methanol, placed in -30 °C refrigerator for 5 h, suction filtration, filter cake with cold methanol washing, 50 °C vacuum drying, yellow solid 96.7 mg, yield 44.70%.

[0152] 1 H NMR (500 MHz, DMSO-d6) δ 12.27 (s, 1H), 10.37 (s, 1H), 7.96 (s, 1H), 7.93-7.89 (m, 2H), 7.82 (s, 1H), 7.76 (d, J = 7.69 Hz, 1H), 7.64 (d, J = 7.74 Hz, 1H), 7.59 (t, J = 7.63 Hz, 1H), 7.43-7.34 (m, 4H), 7.29 (t, J = 7.35 Hz, 1H), 7.02 (s, 1H), 6.94 (d, J = 7.42 Hz, 2H), 6.82 (s, 1H), 5.55 (s, 2H), 1.36 (s, 9H). MS (ESI): m / z 549.78 [M+H] + Mp: 116-118 °C.

[0153] Example 2 Synthesis of (3Z, 6Z)-3-(3-(p-Fluorobenzoyl)phenyl)methylene-6-(5- isopropyl-1-benzylimidazol-4-yl)methylene)piperazine-2, 5-dione (PLN-3-2)

[0154]

[0155] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H- imidazol-4-yl)methylene)piperazine-2, 5-dione (500 mg, 1.81 mmol), potassium carbonate (750.27 mg, 5.43 mmol), DMF (4 ml) in turn, stir for 20 min at room temperature, then add benzyl bromide (619.05 mg, 3.62 mmol), continue to stir the reaction at room temperature for 2.5 h. The reaction solution is dropped into 4 ℃ cold water, solid is precipitated, suction filtration, filter cake is washed with water, vacuum dried at 50 ℃, methanol is pulsed, placed in -30 ℃ refrigerator overnight, suction filtration, filter cake is washed with cold methanol, vacuum dried at 50 ℃, 483.2 mg of brown yellow solid is obtained, the yield is 72.87%.

[0156] 2) Take 25 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1- benzylimidazol-4-yl)methylene)piperazine-2, 5-dione (150 mg, 0.41 mmol), 3-p- fluorobenzoylbenzaldehyde (112.10 mg, 0.49 mmol), cesium carbonate (200.05 mg, 0.61 mmol), anhydrous sodium sulfate (116.29 mg, 0.82 mmol), DMF (3 ml) in turn, exhaust, nitrogen protection, placed in 60 ℃ oil bath, stir for 24 h. LC-MS monitors the reaction, the raw material point disappears. After the reaction is completed, the reaction solution is dropped into 4 ℃ cold water (30 ml), suction filtration, filter cake is washed with cold water (20 ml*2), EA:PE=1:10 (15 ml), the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure, brown solid is obtained, reverse phase column chromatography, gradient elution, 64 mg of light yellow solid is obtained, the yield is 29.24%.

[0157] 1H NMR (500 MHz, DMSO-d6) δ 12.00 (s, 2H), 10.39 (s, 1H), 8.07 (s, 1H), 7.91 (dd, J = 8.5, 5.6 Hz, 2H), 7.84 (s, 1H), 7.77 (d, J = 7.6 Hz, 1H), 7.63 (d, J = 7.7 Hz, 1H), 7.58 (t, J = 7.6 Hz, 1H), 7.39 (dt, J = 12.4, 8.2 Hz, 4H), 7.30 (t, J = 7.3 Hz, 1H), 7.13 (d, J = 7.5 Hz, 2H), 6.81 (s, 1H), 6.67 (s, 1H), 5.35 (s, 2H), 3.12 (dt, J = 14.2, 7.1 Hz, 1H), 1.12 (d, J = 7.1 Hz, 6H). MS (ESI): m / z 535.06 [M+H] + Mp: 216-219 °C.

[0158] Example 3 Preparation of (3Z,6Z)-3-(3-(p-Fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1- benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-3)

[0159]

[0160] A 25 ml dry brown round-bottom flask was taken and (Z)-1-acetyl-3-((5-isopropyl-1- benzylimidazol-4-yl)methylene)piperazine-2,5-dione (141.2 mg, 0.38 mmol), 3-(p- fluorophenylmethoxy)benzaldehyde (100.0 mg, 0.46 mmol), Cs2CO3(188.36 mg, 0.58 mmol), Na2SO4(109.48 mg, 0.77 mmol), DMF (3 ml) were added in sequence, protected by nitrogen, exhausted 3 times, programmed to 60 °C for 24 h. The reaction liquid was poured into cold water (40 ml), a large amount of solid was precipitated, filtered, the filter cake was washed with water, EA:PE = 1:8, washed, methanol and dichloromethane were redissolved. The water phase was extracted with EA, the organic phases were combined, concentrated under reduced pressure, dried. Methanol was pulped (ultrasonic for 20 min), suction filtered, the filter cake was washed with methanol, and dried to obtain 135.3 mg of yellow solid, with a yield of 67.18%. 1 H NMR (400 MHz, DMSO-d6) δ 12.00 (s, 1H), 10.18 (s, 1H), 8.07 (s, 1H), 7.45 - 7.08 (m, 12H), 6.91 (s, 1H), 6.70 (d, J = 21.2 Hz, 2H), 5.35 (s, 2H), 3.12 (s, 1H), 1.11 (s, 6H). 13CNMR (125 MHz, DMSO-d6) δ 159.12, 157.33, 157.22, 157.03, 156.20, 152.57, 138.56, 137.79, 137.08, 135.19, 132.67, 130.27, 128.75, 127.69, 127.20, 126.59, 124.43, 123.99, 120.55, 120.49, 118.91, 117.75, 116.62, 116.43, 113.26, 103.95, 47.97, 23.84, 22.32. MS (ESI): m / z 523.21 [M+H]+. Mp: 184-187 °C.

[0161] Example 4 Synthesis of (3Z,6Z)-3-(3-(p-Fluorobenzoyl)phenyl)methylene-6-(5- isopropyl-l-(2-pyridyl)imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-4)

[0162]

[0163] 1) Take 50 ml dry round bottom flask, add (Z)-l-acetyl-3-((5-isopropyl-lH-imidazol-4- yl)methylene)piperazine-2, 5-dione (300 mg, 1.09 mmol), cesium carbonate (1.42 g, 4.34 mmol), anhydrous sodium sulfate (616.91 mg, 4.34 mmol), DMF (8 ml), stir for 10 min at room temperature, then add 2-(bromomethyl)pyridine hydrobromide (628.53 mg, 2.17 mmol), continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 °C cold water, solid precipitates, suction filtration, filter cake water washing, 50 °C vacuum drying, methanol slurry, placed in -30 °C refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, white solid 326.3 mg, yield 81.79%.

[0164] 2) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(2- pyridyl)imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.41 mmol), 3-p- fluorobenzoylbenzaldehyde (111.90 mg, 0.49 mmol), cesium carbonate (199.66 mg, 0.61 mmol), anhydrous sodium sulfate (116.06 mg, 0.82 mmol), DMF (4 ml), exhaust, nitrogen protection, put into 45 °C oil bath, stir for 18 h. Monitor the reaction by LC-MS, and the starting material point disappears. When the reaction is completed, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtered, and the filter cake is washed with cold water (50 ml*3). The filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, and concentrated under reduced pressure. Ultrasonic slurry with methanol, and place in -30 °C refrigerator for standing, suction filtration, filter cake with cold methanol, 50 °C vacuum drying, get yellow solid 147.6 mg, yield 33.73%.

[0165] 1 H NMR (500 MHz, DMSO-d6) δ 12.01 (s, 1H), 10.36 (s, 1H), 8.53 (d, J = 4.76 Hz, 1H), 8.04 (s, 1H), 7.91 (dd, J = 5.78, 8.17 Hz, 2H), 7.85 - 7.79 (m, 2H), 7.76 (d, J = 7.54 Hz, 1H), 7.64 (d, J = 7.62 Hz, 1H), 7.59 (t, J = 7.63 Hz, 1H), 7.40 (t, J = 8.67 Hz, 2H), 7.33 (dd, J = 4.93, 7.42 Hz, 1H), 7.22 (d, J = 7.82 Hz, 1H), 6.82 (s, 1H), 6.69 (s, 1H), 5.43 (s, 2H), 3.14 (dt, J = 7.10, 14.22 Hz, 1H), 1.13 (d, J = 7.06 Hz, 6H). MS (ESI): m / z 536.17 [M+H] + Mp: 191-193 °C.

[0166] Example 5 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-(5- cyclopropyl-1 -benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-5)

[0167]

[0168] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-cyclopropyl-1H- imidazol-4-yl)methylene)piperazine-2,5-dione (200 mg, 0.73 mmol), potassium carbonate (201.55 mg, 1.46 mmol), DMF (4 ml) in turn, stir at room temperature, then add benzyl bromide (249.44 mg, 1.46 mmol), continue to stir at 40 °C for 1.5 h. The reaction solution is dropped into 4 °C cold water, a solid is precipitated, suction filtration, filter cake water washing, 50 °C vacuum drying, methanol ultrasonic slurry, placed in -30 °C refrigerator overnight, suction filtration, filter cake with cold methanol washing, 50 °C vacuum drying, get brown yellow solid 245 mg, yield 92.20%.

[0169] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-cyclopropyl-1- benzylimidazol-4-yl)methylene)piperazine-2,5-dione (70 mg, 0.19 mmol), 3-para- fluorobenzoyl benzaldehyde (52.60 mg, 0.23 mmol), cesium carbonate (93.87 mg, 0.29 mmol), anhydrous sodium sulfate (54.57 mg, 0.38 mmol), DMF (2 ml) in turn, exhaust, nitrogen protection, placed in 45 °C oil bath, stir for 20 h. LC-MS monitoring reaction, raw material point disappears. Reaction is finished, the reaction solution is dropped into 4 °C cold water (20 ml), suction filtration, filter cake cold water washing (30 ml*2), the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Water phase EA (40 ml*3) extraction until clear, combine organic phase, reduced pressure concentration. Combine filter cake and organic phase sample, methanol ultrasonic slurry, placed in -30 °C refrigerator, suction filtration, filter cake with cold methanol washing, 50 °C vacuum drying oven drying, get yellow solid 76.1 mg, yield 74.38%.

[0170] 1 H NMR (500 MHz, DMSO-d6) δ 11.89 (s, 1H), 10.35 (s, 1H), 8.08 (s, 1H), 7.94-7.87 (m, 2H), 7.81 (s, 1H), 7.75 (d, J = 7.71 Hz, 1H), 7.60 (dt, J = 7.69, 15.27 Hz, 2H), 7.43-7.34 (m, 4H), 7.31 (t, J = 7.32 Hz, 1H), 7.21 (d, J = 7.17 Hz, 2H), 6.81 (s, 1H), 6.69 (s, 1H), 5.34 (s, 2H), 1.48 (tt, J = 5.40, 8.32 Hz, 1H), 1.04-0.90 (m, 2H), 0.68-0.43 (m, 2H). MS (ESI): m / z 533.37 [M+H]+ Mp: 197-200 °C.

[0171] Example 6 Synthesis of (3Z, 6Z)-3-(3-(p-Fluorobenzoyl)phenyl)methylene-6-(5-methyl-1- benzylimidazol-4-yl)methylene)piperazine-2, 5-dione (PLN-3-6)

[0172]

[0173] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-methyl-1H-imidazol-4- yl)methylene)piperazine-2, 5-dione (500 mg, 2.01 mmol), potassium carbonate (835.08 mg, 6.04 mmol), DMF (8 ml) in turn, stir for 20 min at room temperature, then add benzyl bromide (689.02 mg, 4.03 mmol), continue to stir the reaction for 2.5 h at room temperature. The reaction solution is dropped into 4 °C cold water, a solid is precipitated, suction filtration, the filter cake is washed with water, vacuum dried at 50 °C, methanol is pulsed, placed in a -30 °C refrigerator overnight, suction filtration, the filter cake is washed with cold methanol, vacuum dried at 50 °C, 480 mg of brown yellow solid is obtained, the yield is 70.43%.

[0174] 2) Take 25 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-methyl-1-benzylimidazol- 4-yl)methylene)piperazine-2, 5-dione (236 mg, 0.70 mmol), 3-p-fluorobenzoyl benzaldehyde (191.02 mg, 0.84 mmol), cesium carbonate (340.91 mg, 1.05 mmol), anhydrous sodium sulfate (198.15 mg, 1.40 mmol), DMF (4 ml) in turn, exhaust, nitrogen protection, placed in 60 °C oil bath, stir for 24 h. After the reaction is completed, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtration, dissolved in methanol and dichloromethane, filtered, concentrated under reduced pressure, EA is pulsed, placed in a -30 °C refrigerator for 3 h, suction filtration, washed with cold EA, 147.3 mg of yellow solid is obtained, the yield is 41.69%.

[0175] 1H NMR (500 MHz, DMSO-d6) δ 11.87 (s, 1H), 10.36 (s, 1H), 8.11 (s, 1H), 7.91 (dd, J = 8.4, 5.8 Hz, 2H), 7.84 (s, 1H), 7.78 (d, J = 7.5 Hz, 1H), 7.63 (d, J = 7.7 Hz, 1H), 7.58 (t, J = 7.6 Hz, 1H), 7.39 (dd, J = 18.9, 8.3 Hz, 4H), 7.31 (t, J = 7.3 Hz, 1H), 7.18 (d, J = 7.5 Hz, 2H), 6.80 (s, 1H), 6.56 (s, 1H), 5.28 (s, 2H), 2.20 (s, 3H). MS (ESI): m / z 507.10 [M+H] + Mp: 219-221 °C.

[0176] Example 7 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-(5- trifluoromethyl-l-benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-7)

[0177]

[0178] 1) Take 25 mL dry round-bottom flask, add 5-trifluoromethyl-lH-imidazole-4- carboxaldehyde (284 mg, 1.73 mmol), 1,4-diacetylpiperazine-2,5-dione (686.00 mg, 3.46 mmol), cesium carbonate (845.87 mg, 2.60 mmol), DMF (4 mL) in turn, replace nitrogen protection, react at room temperature for 17.5 h. After the reaction, pour the reaction liquid into cold water, a large amount of solid is precipitated, filter, wash the filter cake with cold water, dry, EA, place in -30 °C refrigerator, stand, filter, wash the filter cake with cold EA, dry to get white solid 274 mg, yield 52.38%.

[0179] 2) Take 25 ml dry round bottom flask, add (Z)-1 -acetyl-3-((5-trifluoromethyl- 1 H-imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.50 mmol), potassium carbonate (137.19 mg, 0.99 mmol), anhydrous sodium sulfate (141.00 mg, 0.99 mmol), DMF (2 ml), stir for 20 min at room temperature, then add benzyl bromide (137.19 mg, 0.99 mmol), continue to stir the reaction for 1.5 h at room temperature. Drop the reaction into 4 °C cold water, solid precipitates, suction filtration, filter cake water wash, 50 °C vacuum drying, methanol slurry, placed in -30 °C refrigerator overnight, suction filtration, filter cake with cold methanol wash, 50 °C vacuum drying, white solid 145 mg, yield 74.47%.

[0180] 3) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-trifluoromethyl- 1 -benzylimidazol-4-yl)methylene)piperazine-2,5-dione (100 mg, 0.25 mmol), 3-p- fluorobenzoylbenzaldehyde (69.81 mg, 0.31 mmol), potassium carbonate (52.83 mg, 0.38 mmol), anhydrous sodium sulfate (72.41 mg, 0.51 mmol), DMF (3 ml), exhaust, nitrogen protection, placed in 45 °C oil bath, stir for 10 h. LC-MS monitor the reaction, the reaction is complete, drop the reaction into 4 °C cold water (30 ml), suction filtration, methanol and dichloromethane dissolution, filtration, reduced pressure concentration, methanol slurry, placed in -30 °C refrigerator for 3 h, suction filtration, washed with cold methanol, white solid 129.6 mg, yield 90.71%.

[0181] 1 H NMR (500 MHz, DMSO-d6) δ 11.69 (s, 1H), 10.62 (s, 1H), 8.50 (s, 1H), 7.94 - 7.88 (m, 2H), 7.83 (s, 1H), 7.77 (d, J = 7.59 Hz, 1H), 7.65 (d, J = 7.59 Hz, 1H), 7.59 (t, J = 7.62 Hz, 1H), 7.39 (dd, J = 7.59, 15.64 Hz, 4H), 7.34 (d, J = 6.75 Hz, 1H), 7.18 (d, J = 7.43 Hz, 2H), 6.89 (s, 1H), 6.57 (s, 1H), 5.45 (s, 2H). MS (ESI): m / z 561.41 [M+H] + Mp: 231-232 °C.

[0182] Example 8 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-(l- benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-8)

[0183]

[0184] 1) Take 50 ml dry round bottom flask, add (Z)-1-acetyl-3-((1H-imidazol-4-yl)methylene)piperazine-2,5-dione (600 mg, 2.06 mmol), potassium carbonate (1.06 g, 7.69 mmol), DMF (8 ml) in turn, stir at room temperature, then add benzyl bromide (877.13 mg, 5.13 mmol), continue to stir at room temperature for 2.5 h. The reaction solution is dropped into 4 ℃ cold water, a solid is precipitated, suction filtration, filter cake water washing, 50 ℃ vacuum drying, methanol ultrasonic slurry, placed in-30 ℃ refrigerator overnight, suction filtration, filter cake with cold methanol washing, 50 ℃ vacuum drying, white solid 762 mg, yield 91.63%.

[0185] 2) Take 25 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((1-benzylimidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.46 mmol), 3-p-fluorobenzoyl benzaldehyde (126.66 mg, 0.56 mmol), cesium carbonate (226.02 mg, 0.69 mmol), anhydrous sodium sulfate (131.38 mg, 0.92 mmol), DMF (3 ml) in turn, exhaust, nitrogen protection, placed in 45 ℃ oil bath, stir for 25 h. LC-MS monitoring reaction, reaction is completed, the reaction solution is dropped into 4 ℃ cold water (40 ml), suction filtration, filter cake cold water (30 ml*3) washing, suction dry, methanol and dichloromethane (1:3) dissolution, filtration, reduced pressure concentration, methanol ultrasonic slurry, to the-30 ℃ under overnight, suction filtration, filter cake with cold methanol washing, 50 ℃ vacuum drying, yellow solid 109.2 mg, yield 47.94%.

[0186] 1 1H NMR (500 MHz, DMSO-d6) δ 11.75 (s, 1H), 10.34 (s, 1H), 8.13 (s, 1H), 7.95-7.88 (m, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.15 Hz, 1H), 7.66-7.55 (m, 3H), 7.35 (ddd, J = 7.87, 16.01, 22.93 Hz, 7H), 6.82 (s, 1H), 6.61 (s, 1H), 5.28 (s, 2H). MS (ESI): m / z 493.10 [M+H] +Mp: 186-188 °C.

[0187] Example 9 Synthesis of (3Z,6Z)-3-phenylmethylene-6-((5-tert-butyl-l- benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-9)

[0188]

[0189] A 25 ml dry brown round-bottom flask was charged with (Z)-l-acetyl-3-((5-tert- butyl-l-benzylimidazol-4-yl)methylene)piperazine-2,5-dione (185 mg, 0.49 mmol), benzaldehyde (61.92 mg, 0.58 mmol), cesium carbonate (237.65 mg, 0.73 mmol), anhydrous sodium sulfate (138.14 mg, 0.97 mmol), DMF (3 ml), evacuated, nitrogen protection, placed in a 50 °C oil bath, and stirred to react for 24 h in the dark. After the reaction, the reaction solution was dropped into 4 °C cold water (30 ml), suction filtered, and the filter cake was washed with cold water (20 ml*3). The filter cake was dissolved in a mixture of methanol and dichloromethane (1:3), filtered, and concentrated under reduced pressure. The aqueous phase was extracted with EA until it was clear, the organic phases were combined, and concentrated under reduced pressure. The filter cake and organic phase sample were combined, methanol was ultrasonically beaten, placed in a -30 °C refrigerator for 5 h, suction filtered, the filter cake was washed with cold methanol, and dried at 50 °C under vacuum to obtain 169.6 mg of yellow solid with a yield of 81.77%.

[0190] 1 H NMR (500 MHz, DMSO-d6) δ 12.24 (s, 1H), 10.06 (s, 1H), 7.96 (s, 1H), 7.53 (d, J = 7.54 Hz, 2H), 7.42 (t, J = 7.70 Hz, 2H), 7.37 (t, J = 7.56 Hz, 2H), 7.30 (dt, J = 7.39, 17.80 Hz, 2H), 7.02 (s, 1H), 6.94 (d, J = 7.35 Hz, 2H), 6.76 (s, 1H), 5.55 (s, 2H), 1.37 (s, 9H). MS (ESI): m / z 427.12 [M+H] + Mp: 220-223 °C.

[0191] Example 10 Synthesis of (3Z,6Z)-3-phenylmethylene-6-((5-isopropyl-l- benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-10)

[0192]

[0193] Into a 25 ml dry brown round-bottom flask, (Z)-1 -acetyl-3-((5-isopropyl-1 -benzylimidazol-4-yl)methyl)methyl)piperazine-2,5-dione (172 mg, 0.47 mmol), benzaldehyde (59.77 mg, 0.56 mmol), cesium carbonate (229.41 mg, 0.70 mmol), anhydrous sodium sulfate (133.35 mg, 0.94 mmol), DMF (3 ml), exhaust, nitrogen protection, placed in 55 °C oil bath, dark stirring reaction 20 h. LC-MS monitoring reaction, raw material point disappeared. Reaction, the reaction liquid drop into 4 °C cold water (30 ml), there is a yellow solid precipitation, suction filtration, filter cake cold water wash (30 ml*2), EA: PE = 1:10 (20 ml) wash, the filter cake was dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration, EA beating, in the-30 °C refrigerator overnight, suction filtration, filter cake cold EA wash, 50 °C vacuum drying of yellow solid 111.2 mg, yield 57.43%.

[0194] 1 H NMR (500 MHz, DMSO-d6) δ 11.97 (s, 1H), 10.07 (s, 1H), 8.06 (s, 1H), 7.53 (d, J = 7.7 Hz, 2H), 7.39 (dt, J = 21.0, 7.6 Hz, 4H), 7.30 (q, J = 7.5 Hz, 2H), 7.13 (d, J = 7.5 Hz, 2H), 6.76 (s, 1H), 6.68 (s, 1H), 5.34 (s, 2H), 3.12 (dq, J = 14.2, 7.1 Hz, 1H), 1.12 (d, J = 7.1 Hz, 6H). MS (ESI): m / z 413.10 [M+H] + Mp: 231-233 °C.

[0195] Example 11 Synthesis of (3Z,6Z)-3-benzylidene-6-((5-cyclopropyl-1 -benzylimidazol-4- yl)methyl)piperazine-2,5-dione (PLN-3-11)

[0196]

[0197] Take 10 ml dry brown round-bottom flask, add (Z)-1-acetyl-3-((5-cyclopropyl-1- benzylimidazol-4-yl)methylene)piperazine-2,5-dione (70 mg, 0.19 mmol), benzaldehyde (24.46 mg, 0.23 mmol), cesium carbonate (93.87 mg, 0.29 mmol), anhydrous sodium sulfate (54.57 mg, 0.38 mmol), DMF (2 ml), exhaust, nitrogen protection, placed in 45 °C oil bath, dark stirring reaction 24 h. After the reaction, the reaction liquid was dropped into 4 °C cold water (30 ml), suction filtration, filter cake cold water wash (30 ml*3), the filter cake was dissolved in methanol and dichloromethane (1:3) mixed solution, filtered, reduced pressure concentration. The water phase EA (40 ml*3) was extracted to clear, the organic phase was combined, and reduced pressure concentration. The filter cake and organic phase sample were combined, methanol was ultrasonically beaten, and placed in a-30 °C refrigerator for standing, suction filtration, filter cake was washed with cold methanol, and 50 °C vacuum drying, 60.7 mg of yellow solid was obtained, the yield was 76.98%.

[0198] 1 H NMR (500 MHz, DMSO-d6) δ 11.87 (s, 1H), 10.05 (s, 1H), 8.09 (s, 1H), 7.53 (d, J = 7.53 Hz, 2H), 7.40 (dt, J = 7.59, 18.94 Hz, 4H), 7.34 - 7.29 (m, 2H), 7.22 (d, J = 7.28 Hz, 2H), 6.76 (s, 1H), 6.70 (s, 1H), 5.34 (s, 2H), 1.49 (tt, J = 5.42, 8.29 Hz, 1H), 1.04 - 0.90 (m, 2H), 0.72 - 0.45 (m, 2H). MS (ESI): m / z 411.09 [M + H] + Mp: 246-249 °C.

[0199] Example 12 Synthesis of (3Z,6Z)-3-benzylidene-6-((5-methyl-1-benzylimidazol-4- yl)methylene)piperazine-2,5-dione (PLN-3-12)

[0200]

[0201] Into a 25 ml dry brown round-bottom flask, (Z)-1-acetyl-3-((5-methyl-1- benzylimidazol-4-yl)methylidene)piperazine-2,5-dione (197.80 mg, 0.58 mmol), benzaldehyde (74.44 mg, 0.70 mmol), cesium carbonate (285.71 mg, 0.88 mmol), anhydrous sodium sulfate (166.06 mg, 1.17 mmol), DMF (3 ml), exhaust, nitrogen protection, placed in 60 °C oil bath, dark stirring reaction for 24 h. After the reaction, the reaction liquid was dropped into 4 °C cold water (30 ml), suction filtration, filter cake cold water wash (20 ml*3), EA:PE = 1:10 (20 ml) wash, the filter cake was dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure, EA beating, in the-30 °C refrigerator overnight, suction filtration, filter cake cold EA wash, 50 °C vacuum drying to get yellow solid 140 mg, yield 62.29%.

[0202] 1 H NMR (500 MHz, DMSO-d6) δ 11.86 (s, 1H), 10.02 (s, 1H), 8.11 (s, 1H), 7.53 (d, J = 7.6 Hz, 2H), 7.41 (dd, J = 12.9, 5.0 Hz, 2H), 7.37 (d, J = 7.7 Hz, 2H), 7.34 - 7.28 (m, 2H), 7.18 (d, J = 7.2 Hz, 2H), 6.76 (s, 1H), 6.57 (s, 1H), 5.27 (d, J = 14.2 Hz, 2H), 2.20 (s, 3H). MS (ESI): m / z 385.05 [M+H] + Mp: 236-238 °C.

[0203] Example 13 Synthesis of (3Z,6Z)-3-benzylidene-6-((1-benzylimidazol-4-yl)methylidene)piperazine-2,5-dione (PLN-3-13)

[0204]

[0205] Take 25 ml dry brown round-bottom flask, add (Z)-1-acetyl-3-((1-benzylimidazol-4- yl)methylene)piperazine-2,5-dione (150 mg, 0.46 mmol), benzaldehyde (58.90 mg, 0.56 mmol), cesium carbonate (226.02 mg, 0.69 mmol), anhydrous sodium sulfate (131.38 mg, 0.97 mmol), DMF (3 ml) in turn, exhaust, nitrogen protection, place in 50 ℃ oil bath, stir the reaction for 24 h in the dark. After the reaction, the reaction liquid was dropped into 4 ℃ cold water (40 ml), suction filtration, filter cake cold water wash (30 ml*3), the filter cake was dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure. The water phase was extracted with EA until clear, the organic phases were combined and concentrated under reduced pressure. The filter cake and organic phase sample were combined, methanol was ultrasonically beaten, placed in a-30 ℃ refrigerator, suction filtration, filter cake with cold methanol wash, 50 ℃ vacuum drying, 106.2 mg of yellow solid was obtained, the yield was 61.99%.

[0206] 1 H NMR (500 MHz, DMSO-d6) δ 11.75 (s, 1H), 10.04 (s, 1H), 8.14 (s, 1H), 7.63 (d, J = 0.86 Hz, 1H), 7.54 (d, J = 7.60 Hz, 2H), 7.45 - 7.37 (m, 4H), 7.35 - 7.28 (m, 4H), 6.78 (s, 1H), 6.62 (s, 1H), 5.29 (s, 2H). MS (ESI): m / z 371.00 [M+H] + Mp: 204-206 °C.

[0207] Example 14 Synthesis of (3Z,6Z)-3-(2,5-difluorophenyl)methylene-6-((5-tert-butyl-1- benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-14)

[0208]

[0209] Take 10 ml dry brown round-bottom flask, add (Z)-1 -acetyl-3-((5-tert-butyl 1 - benzylimidazol-4-yl)methylene)piperazine-2, 5-dione (150 mg, 0.39 mmol), 2,5- difluorobenzaldehyde (67.23 mg, 0.47 mmol), cesium carbonate (192.69 mg, 0.59 mmol), anhydrous sodium sulfate (112.13 mg, 0.79 mmol), DMF (3 ml), exhaust, nitrogen protection, placed in 45 °C oil bath, dark stirring reaction for 18 h. LC-MS monitoring reaction, reaction, drop the reaction liquid into 4 °C cold water (40 ml), suction filtration, filter cake cold water wash (30 ml*3), suction dry, the filter cake was dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Water phase EA (50 ml*2) extraction to clear, combined organic phase, reduced pressure concentration. Combined filter cake and organic phase sample, methanol ultrasonic slurry, placed in-30 °C refrigerator, suction filtration, filter cake with cold methanol wash, 50 °C vacuum drying, get yellow solid 157.8 mg, yield 86.53%.

[0210] 1 H NMR (500 MHz, DMSO-d6) δ 12.31 (s, 1H), 10.48 (s, 1H), 7.97 (s, 1H), 7.48 - 7.34 (m, 3H), 7.29 (td, J = 4.96, 9.64 Hz, 2H), 7.24 - 7.16 (m, 1H), 7.05 (s, 1H), 6.94 (d, J = 7.38 Hz, 2H), 6.63 (s, 1H), 5.56 (s, 2H), 1.37 (s, 9H). MS (ESI): m / z 463.15 [M+H] + Mp: 196-198 °C.

[0211] Example 15 Synthesis of (3Z,6Z)-3-(2,5-difluorophenyl)methylene-6-((5-isopropyl-1- benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-15)

[0212]

[0213] Take 25 ml dry brown round-bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 - benzylimidazol-4-yl)methylene)piperazine-2, 5-dione (200 mg, 0.55 mmol), 2, 5- difluorobenzaldehyde (93.08 mg, 0.66 mmol), cesium carbonate (266.75 mg, 0.82 mmol), anhydrous sodium sulfate (155.06 mg, 1.09 mmol), DMF (4 ml), exhaust, nitrogen protection, placed in 50 °C oil bath, dark stirring reaction for 18 h. LC-MS monitoring reaction, reaction, drop the reaction liquid into 4 °C cold water (40 ml), suction filtration, filter cake cold water wash (30 ml*3), dry suction, the filter cake was dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Water phase EA extraction to clear, combined organic phase, reduced pressure concentration. Combined filter cake and organic phase sample, methanol ultrasonic slurry, placed in -30 °C refrigerator, suction filtration, filter cake cold methanol wash, 50 °C vacuum drying, get yellow solid 157.8 mg, yield 62.51%.

[0214] 1 H NMR (500 MHz, DMSO-d6) δ 12.05 (s, 1H), 10.47 (s, 1H), 8.07 (s, 1H), 7.40 (ddd, J = 5.41, 10.31, 20.45 Hz, 3H), 7.33 - 7.26 (m, 2H), 7.23 - 7.17 (m, 1H), 7.13 (d, J = 7.37 Hz, 2H), 6.70 (s, 1H), 6.63 (s, 1H), 5.35 (s, 2H), 3.12 (dt, J = 7.11, 14.24 Hz, 1H), 1.12 (d, J = 7.11 Hz, 6H). MS (ESI): m / z 449.11 [M+H] + . Mp: 205-207 °C.

[0215] Example 16 Synthesis of (3Z,6Z)-3-(2,5-difluorophenyl)methylene-6-((5-cyclopropyl-1- benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-16)

[0216]

[0217] Take 10 ml dry brown round-bottom flask, add (Z)-1 -acetyl-3-((5-cyclopropyl 1 - benzylimidazol-4-yl)methylene)piperazine-2, 5-dione (70 mg, 0.19 mmol), 2,5-difluorobenzaldehyde (32.75 mg, 0.23 mmol), cesium carbonate (93.87 mg, 0.29 mmol), anhydrous sodium sulfate (54.57 mg, 0.38 mmol), DMF (2 ml), exhaust, nitrogen protection, placed in 45 °C oil bath, dark stirring reaction 21 h. LC-MS monitoring reaction, reaction, drop the reaction liquid into 4 °C cold water (20 ml), suction filtration, filter cake cold water wash (20 ml*3), dry suction, the filter cake was dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Water phase EA (30 ml*3) extraction to clear, combined organic phase, reduced pressure concentration. Combined filter cake and organic phase sample, methanol ultrasonic slurry, placed in -30 °C refrigerator, suction filtration, filter cake with cold methanol wash, 50 °C vacuum drying, get yellow solid 55.8 mg, yield 65.07%.

[0218] 1 H NMR (500 MHz, DMSO-d6) δ 11.93 (s, 1H), 10.47 (s, 1H), 8.09 (s, 1H), 7.39 (ddd, J = 5.40, 10.22, 19.64 Hz, 3H), 7.34 - 7.24 (m, 2H), 7.24 - 7.16 (m, 3H), 6.72 (s, 1H), 6.62 (s, 1H), 5.34 (s, 2H), 1.49 (tt, J = 5.40, 8.29 Hz, 1H), 1.01 - 0.88 (m, 2H), 0.70 - 0.45 (m, 2H). MS (ESI): m / z 447.10 [M+H] + .Mp: 228-229 °C.

[0219] Example 17 Synthesis of (3Z,6Z)-3-(2,5-difluorophenyl)methylene-6-((5-methyl-1- benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-17)

[0220]

[0221] Into a 25 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1- benzylimidazol-4-yl)methyl)methyl)piperazine-2,5-dione (200 mg, 0.59 mmol), 2,5- difluorobenzaldehyde (100.79 mg, 0.71 mmol), cesium carbonate (288.87 mg, 0.89 mmol), anhydrous sodium sulfate (167.91 mg, 1.18 mmol), DMF (4 ml), exhaust, nitrogen protection, placed in 50 °C oil bath, dark stirring reaction for 24 h. After the reaction, the reaction liquid was dropped into 4 °C cold water (40 ml), suction filtration, filter cake cold water wash (30 ml*3), dry suction, the filter cake was dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure, EA, in the-30 °C refrigerator overnight, suction filtration, filter cake cold EA wash, 50 °C vacuum drying to get yellow solid 176.7 mg, yield 71.10%.

[0222] 1 H NMR (500 MHz, DMSO-d6) δ 11.92 (s, 1H), 10.45 (s, 1H), 8.12 (s, 1H), 7.31 (ddd, J = 18.78, 45.59, 66.30 Hz, 6H), 6.61 (d, J = 15.76 Hz, 2H), 5.29 (s, 2H), 2.50 (s, 3H). MS (ESI): m / z 421.06 [M+H] + .Mp: 180-182℃.

[0223] Example 18 Synthesis of (3Z,6Z)-3-(2,5-difluorophenyl)methyl)-6-((1-benzylimidazol-4- yl)methyl)piperazine-2,5-dione (PLN-3-18)

[0224]

[0225] Take 25 ml dry brown round-bottom flask, add (Z)-1 -acetyl-3-((1 -benzylimidazol-4- yl)methyl)methyl)piperazine-2, 5-dione (150 mg, 0.46 mmol), 2, 5-difluorobenzaldehyde (7.86 mg, 0.56 mmol), cesium carbonate (226.02 mg, 0.69 mmol), anhydrous sodium sulfate (131.38 mg, 0.92 mmol), DMF (3 ml), exhaust, nitrogen protection, placed in 45 °C oil bath, dark stirring reaction 19 h. LC-MS monitoring reaction, reaction, drop the reaction liquid into 4 °C cold water (40 ml), suction filtration, filter cake cold water wash (30 ml*3), dry suction, the filter cake was dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Water phase EA (50 ml*2) extraction to clear, combined organic phase, reduced pressure concentration. Combined filter cake and organic phase sample, methanol ultrasonic slurry, placed in-30 °C refrigerator, suction filtration, filter cake with cold methanol wash, 50 °C vacuum drying, get yellow solid 97.4 mg, yield 51.82%.

[0226] 1 H NMR (500 MHz, DMSO-d6) δ 11.81 (s, 1H), 10.45 (s, 1H), 8.14 (s, 1H), 7.64 (s, 1H), 7.39 (dd, J = 7.16, 14.38 Hz, 3H), 7.36 - 7.25 (m, 4H), 7.24 - 7.16 (m, 1H), 6.64 (d, J = 2.97 Hz, 2H), 5.28 (s, 2H). MS (ESI): m / z 407.03 [M+H] + .Mp:201-205℃.

[0227] Example 19 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methyl-6-(5-isopropyl-1-(4- fluoro)benzylimidazol-4-yl)methyl)piperazine-2, 5-dione (PLN-3-19)

[0228]

[0229] 1) Take 25 ml dry round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 H- imidazol-4-yl)methylene)piperazine-2,5-dione (200 mg, 0.72 mmol), potassium carbonate (300.13 mg, 2.18 mmol), DMF (4 ml) successively, stir at room temperature for 20 min, then add 4-fluorobenzyl bromide (247.63 mg, 1.45 mmol), continue to stir at room temperature for 2 h. Drop the reaction solution into 4 °C cold water, solid precipitates, suction filtration, filter cake water washing, 50 °C drying, methanol slurry, placed in -30 °C refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get brown yellow solid 252 mg, yield 95.00%.

[0230] 2) Take 25 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(4- fluoro)benzyl imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.39 mmol), 3- p-fluorobenzoyl benzaldehyde (106.88 mg, 0.47 mmol), cesium carbonate (190.70 mg, 0.58 mmol), anhydrous sodium sulfate (110.85 mg, 0.78 mmol), DMF (4 ml) successively, exhaust, nitrogen protection, placed in 45 °C oil bath, stir for 18 h. LC-MS monitoring reaction, raw material point disappears. Reaction is completed, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtration, filter cake cold water washing, the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Methanol ultrasonic slurry, placed in -30 °C refrigerator, suction filtration, filter cake with cold methanol washing, 50 °C vacuum drying, get yellow solid 106.0 mg, yield 49.16%.

[0231] 1 H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.37 (s, 1H), 8.07 (s, 1H), 7.93-7.88 (m, 2H), 7.82 (s, 1H), 7.76 (d, J = 7.54 Hz, 1H), 7.64 (d, J = 7.68 Hz, 1H), 7.59 (t, J = 7.62 Hz, 1H), 7.40 (t, J = 8.73 Hz, 2H), 7.21 (d, J = 7.05 Hz, 4H), 6.82 (s, 1H), 6.68 (s, 1H), 5.33 (s, 2H), 3.13 (dt, J = 7.11, 14.24 Hz, 1H). 1.13 (d, J = 7.04 Hz, 6H). MS (ESI): m / z 553.06 [M+H] + Mp: 172-176 °C.

[0232] Example 20 Synthesis of (3Z, 6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-(5-isopropyl- 1-(3-fluoro)benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-20)

[0233]

[0234] A 25 ml dry brown round bottom flask was charged with (Z)-1-acetyl-3-((5-isopropyl- 1-(3-fluoro)benzylimidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.39 mmol), 3-p-fluorobenzoylbenzaldehyde (106.88 mg, 0.47 mmol), cesium carbonate (190.70 mg, 0.58 mmol), anhydrous sodium sulfate (110.85 mg, 0.78 mmol), DMF (4 ml), evacuated, nitrogen protection, placed in a 45 °C oil bath, and stirred for 14 h. The reaction was monitored by LC-MS, and the starting material point disappeared. After the reaction was completed, the reaction solution was dropped into 4 °C cold water (40 ml), suction filtered, and the filter cake was washed with cold water. The filter cake was dissolved in a mixture of methanol and dichloromethane (1:3), filtered, and concentrated under reduced pressure. The methanol was ultrasonically slurried, placed in a -30 °C refrigerator, suction filtered, and the filter cake was washed with cold methanol. The filter cake was dried at 50 °C under vacuum to give 116.4 mg of a yellow solid with a yield of 53.98%.

[0235] 1 H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.37 (s, 1H), 8.08 (s, 1H), 7.95-7.88 (m, 2H), 7.82 (s, 1H), 7.76 (d, J = 7.57 Hz, 1H), 7.64 (d, J = 7.51 Hz, 1H), 7.59 (t, J = 7.61 Hz, 1H), 7.46-7.37 (m, 3H), 7.15 (t, J = 8.47 Hz, 1H), 7.01 (d, J = 9.78 Hz, 1H), 6.94 (d, J = 7.65 Hz, 1H), 6.82 (s, 1H), 6.68 (s, 1H), 5.37 (s, 2H), 3.12 (dt, J = 6.95, 13.97 Hz, 1H), 1.13 (d, J = 6.91 Hz, 6H). MS (ESI): m / z 553.00 [M+H] + Mp: 182-183 °C.

[0236] Example 21 Synthesis of (3Z, 6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-(5-isopropyl- 1-(2-fluoro)benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-21)

[0237]

[0238] Into a 25 ml dry brown round bottom flask, (Z)-1 -acetyl-3-((5-isopropyl-1 -(2- fluoro)benzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.39 mmol), 3- p-toluoylbenzaldehyde (106.88 mg, 0.47 mmol), cesium carbonate (190.70 mg, 0.58 mmol), anhydrous sodium sulfate (110.85 mg, 0.78 mmol), DMF (4 ml), evacuate, nitrogen protection, put into 45 °C oil bath, stir for 18 h. Monitor the reaction by LC-MS, the raw material point disappears. When the reaction is completed, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtration, the filter cake is washed with cold water, the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure. Ultrasonic slurry with methanol, placed in -30 °C refrigerator, suction filtration, the filter cake is washed with cold methanol, vacuum dried at 50 °C, to get yellow solid 145.6 mg, yield 67.53%.

[0239] 1 H NMR (500 MHz, DMSO-d6) δ 11.97 (s, 1H), 10.37 (s, 1H), 8.01 (s, 1H), 7.94 - 7.89 (m, 2H), 7.82 (s, 1H), 7.76 (d, J = 7.52 Hz, 1H), 7.64 (d, J = 7.57 Hz, 1H), 7.59 (t, J = 7.58 Hz, 1H), 7.40 (t, J = 8.37 Hz, 3H), 7.31 - 7.24 (m, 1H), 7.21 (t, J = 7.50 Hz, 1H), 7.01 (t, J = 7.64 Hz, 1H), 6.82 (s, 1H), 6.69 (s, 1H), 5.40 (s, 2H), 3.14 (dt, J = 7.03, 14.09 Hz, 1H), 1.16 (d, J = 6.94 Hz, 6H). MS (ESI): m / z 553.34 [M+H] + Mp: 178-180 °C.

[0240] Example 22 Preparation of (3Z,6Z)-3-(3-(p-toluoyl)benzylidene)-6-((5-isopropyl-1- p-chlorobenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-22)

[0241]

[0242] 1) Take 25 ml dry round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 H- imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), 4-chlorobenzyl bromide (446.22 mg, 2.17 mmol) in DMF (6 ml) successively, continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, a solid is precipitated, suction filtration, the filter cake is washed with water, dried in a vacuum drying oven at 50 ℃, methanol is pulped, placed in a refrigerator at -30 ℃ for more than 2 h, suction filtration, washed with cold methanol, dried, and white solid 314.3 mg is obtained with a yield of 72.21%.

[0243] 2) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(4- chloro)benzyl imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.37 mmol), 3- p-fluorobenzoyl benzaldehyde (102.47 mg, 0.45 mmol), cesium carbonate (182.88 mg, 0.56 mmol), anhydrous sodium sulfate (106.30 mg, 0.75 mmol), DMF (4 ml) successively, exhaust, nitrogen protection, place in 45 ℃ oil bath, stir for 13.5 h. Monitor the reaction by LC-MS, and the raw material point disappears. After the reaction is completed, the reaction solution is dropped into 4 ℃ cold water (40 ml), suction filtration, the filter cake is washed with cold water, the filter cake is dissolved in a mixed solution of methanol and dichloromethane (1:3), filtered, and concentrated under reduced pressure. Methanol is ultrasonically pulped, placed in a refrigerator at -30 ℃, suction filtered, the filter cake is washed with cold methanol, and dried in a vacuum drying oven at 50 ℃ to obtain yellow solid 173.0 mg with a yield of 81.25%.

[0244] 1 H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.37 (s, 1H), 8.07 (s, 1H), 7.91 (s, 2H), 7.82 (s, 1H), 7.76 (d, J = 6.44 Hz, 1H), 7.67 - 7.55 (m, 2H), 7.43 (dd, J = 8.16, 17.35 Hz, 4H), 7.16 (d, J = 7.47 Hz, 2H), 6.82 (s, 1H), 6.68 (s, 1H), 5.35 (s, 2H), 3.11 (d, J = 6.37 Hz, 1H), 1.13 (d, J = 6.45 Hz, 6H). MS (ESI): m / z 570.31 [M+H] + Mp: 174-176 ℃.

[0245] Example 23 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-m-chlorobenzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-23)

[0246]

[0247] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.06 mmol), potassium carbonate (450.16 mg, 3.26 mmol), 3-chlorobenzyl bromide (446.22 mg, 2.1716 mmol) in DMF (6 ml) successively, continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, a solid is precipitated, suction filtration, the filter cake is washed with water, 50 ℃ vacuum drying, methanol slurry, placed in the-30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get white solid 330.9 mg, yield 76.03%.

[0248] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1-(3-chloro)benzyl imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.37 mmol), 3-p-fluorobenzoyl benzaldehyde (102.47 mg, 0.45 mmol), cesium carbonate (182.88 mg, 0.56 mmol), anhydrous sodium sulfate (106.30 mg, 0.75 mmol), DMF (3.5 ml) successively, exhaust, nitrogen protection, placed in 45 ℃ oil bath, stirring reaction 13.5 h. LC-MS monitoring reaction, raw material point disappears. After the reaction, the reaction solution is dropped into 4 ℃ cold water (40 ml), suction filtration, the filter cake is washed with cold water, the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure. Methanol ultrasonic slurry, placed in-30 ℃ refrigerator, suction filtration, the filter cake is washed with cold methanol, 50 ℃ vacuum drying, get yellow solid 131.0 mg, yield 61.52%.

[0249] 1 H NMR (500 MHz, DMSO-d6) δ 11.98 (s, 1H), 10.37 (s, 1H), 8.08 (s, 1H), 7.97-7.71 (m, 4H), 7.63 (s, 2H), 7.40 (s, 4H), 7.23 (s, 1H), 7.07 (s, 1H), 6.82 (s, 1H), 6.68 (s, 1H), 5.37 (s, 2H), 3.12 (s, 1H), 1.14 (s, 6H). MS (ESI): m / z 570.53 [M+H]+ Mp: 254-256 °C.

[0250] Example 24. Preparation of (3Z, 6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5- isopropyl-1-o-chlorobenzylimidazol-4-yl)methylene)piperazine-2, 5-dione (PLN-3-24)

[0251]

[0252] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2, 5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), 2-chlorobenzyl bromide (446.22 mg, 2.17 mmol) in DMF (6 ml) successively, continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 °C cold water, solid precipitates, suction filtration, filter cake water washing, 50 °C vacuum drying, methanol slurry, placed in -30 °C refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, to get white solid 362.5 mg, yield 83.29%.

[0253] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1-(2-chloro)benzyl imidazol-4-yl)methylene)piperazine-2, 5-dione (150 mg, 0.37 mmol), 3-p-fluorobenzoyl benzaldehyde (102.47 mg, 0.45 mmol), cesium carbonate (182.88 mg, 0.56 mmol), anhydrous sodium sulfate (106.30 mg, 0.75 mmol), DMF (3.5 ml) successively, exhaust, nitrogen protection, placed in 45 °C oil bath, stirring reaction for 13.0 h. LC-MS monitoring reaction, raw material point disappears. After the reaction, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtration, filter cake cold water washing, the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Methanol ultrasonic slurry, placed in -30 °C refrigerator, suction filtration, filter cake with cold methanol washing, 50 °C vacuum drying, to get yellow solid 127.4 mg, yield 59.84%.

[0254] 1H NMR (500 MHz, DMSO-d6) δ 11.98 (s, 1H), 10.38 (s, 1H), 7.99 (s, 1H), 7.95 - 7.88 (m, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.26 Hz, 1H), 7.66 - 7.52 (m, 3H), 7.45 - 7.32 (m, 4H), 6.83 (s, 1H), 6.71 (s, 2H), 5.41 (s, 2H), 3.04 (dt, J = 6.84, 14.04 Hz, 1H), 1.18 (d, J = 6.93 Hz, 6H). MS (ESI): m / z 570.16 [M+H] + Mp: 179-181 °C.

[0255] Example 25 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-p-bromobenzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-25)

[0256]

[0257] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2, 5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), 4-bromobenzyl bromide (542.75 mg, 2.17 mmol) in DMF (6 ml) in turn, continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, solid is precipitated, suction filtration, filter cake is washed with water, 50 ℃ vacuum drying, methanol is pulsed, placed in-30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get white solid 372.4 mg, yield 77.02%.

[0258] 2) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(4- bromo)benzyl imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.34 mmol), 3-p- fluorobenzoylbenzaldehyde (92.25 mg, 0.40 mmol), cesium carbonate (164.64 mg, 0.50 mmol), anhydrous sodium sulfate (95.69 mg, 0.67 mmol), DMF (3.5 ml), evacuate, nitrogen protection, put into 45 °C oil bath, stir the reaction for 15.5 h. LC-MS monitor the reaction, the raw material point disappears. When the reaction is completed, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtration, the filter cake is washed with cold water, the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure, and dried. Ultrasonic slurry in methanol, placed in -30 °C refrigerator, suction filtration, the filter cake is washed with cold methanol, vacuum dried at 50 °C, to get yellow solid 162.7 mg, yield 78.74%.

[0259] 1 H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.41 (s, 1H), 8.08 (s, 1H), 7.94 - 7.88 (m, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.47 Hz, 1H), 7.64 (d, J = 7.57 Hz, 1H), 7.59 (t, J = 7.32 Hz, 3H), 7.40 (t, J = 8.22 Hz, 2H), 7.09 (d, J = 7.68 Hz, 2H), 6.82 (s, 1H), 6.67 (s, 1H), 5.34 (s, 2H), 3.09 (dt, J = 7.05, 14.04 Hz, 1H), 1.12 (d, J = 6.92 Hz, 6H). MS (ESI): m / z 614.89 [M+H] + Mp: 154-155 °C.

[0260] Example 26 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-m-bromobenzyl imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-26)

[0261]

[0262] 1) Take 25 ml dry round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 H- imidazol-4-yl)methyl)piperazine-2,5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), 3-bromobenzyl bromide (542.75 mg, 2.17 mmol) in DMF (6 ml) successively, continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, a solid is precipitated, suction filtration, the filter cake is washed with water, 50 ℃ vacuum drying, methanol slurry, placed in the refrigerator at -30 ℃ for more than 2 h, suction filtration, washed with cold methanol, dried, to get a white solid 368.7 mg, yield 76.25%.

[0263] 2) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(3- bromo)benzyl imidazol-4-yl)methyl)piperazine-2,5-dione (150 mg, 0.34 mmol), 3- p-fluorobenzoyl benzaldehyde (92.25 mg, 0.40 mmol), cesium carbonate (164.64 mg, 0.50 mmol), anhydrous sodium sulfate (95.69 mg, 0.67 mmol), DMF (3.5 ml), exhaust, nitrogen protection, placed in 45 ℃ oil bath, stirring reaction for 24 h. LC-MS monitoring reaction, raw material point disappears. After the reaction, the reaction solution is dropped into 4 ℃ cold water (40 ml), suction filtration, the filter cake is washed with cold water, the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure, dried. Methanol ultrasonic slurry, placed in the refrigerator at -30 ℃, suction filtration, the filter cake is washed with cold methanol, 50 ℃ vacuum drying, to get a yellow solid 121.1 mg, yield 58.61%.

[0264] 1 H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.41 (s, 1H), 8.09 (s, 1H), 7.94-7.88 (m, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.45 Hz, 1H), 7.64 (d, J = 7.55 Hz, 1H), 7.58 (t, J = 7.53 Hz, 1H), 7.51 (d, J = 7.93 Hz, 1H), 7.44-7.30 (m, 4H), 7.10 (d, J = 7.70 Hz, 1H), 6.82 (s, 1H), 6.67 (s, 1H), 5.37 (s, 2H), 3.11 (dt, J = 6.86, 13.98 Hz, 1H), 1.13 (d, J = 6.87 Hz, 6H). MS (ESI): m / z 614.63 [M+H] + Mp: 174-176 ℃.

[0265] Example 27 Preparation of (3Z, 6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-o-bromobenzylimidazol-4-yl)methylene)piperazine-2, 5-dione (PLN-3-27)

[0266]

[0267] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2, 5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), 2-bromobenzyl bromide (254.75 mg, 2.17 mmol) in DMF (6 ml) successively, continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, a solid is precipitated, suction filtration, the filter cake is washed with water, dried at 50 ℃ under vacuum, methanol is pulsed, placed in a refrigerator at -30 ℃ for more than 2 h, suction filtration, washed with cold methanol, dried, to get brown yellow solid 391.1 mg, yield 80.88%.

[0268] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1-(2-bromo)benzylimidazol-4-yl)methylene)piperazine-2, 5-dione (150 mg, 0.34 mmol), 3-p-fluorobenzoyl benzaldehyde (92.25 mg, 0.40 mmol), cesium carbonate (164.64 mg, 0.50 mmol), anhydrous sodium sulfate (95.69 mg, 0.67 mmol), DMF (3.5 ml) successively, exhaust, nitrogen protection, placed in 45 ℃ oil bath, stir the reaction for 18 h. LC-MS monitoring reaction, raw material point disappears. After the reaction, the reaction solution is dropped into 4 ℃ cold water (40 ml), suction filtration, the filter cake is washed with cold water, the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure, dried. Methanol is pulsed with ultrasonic, placed in a refrigerator at -30 ℃, suction filtration, the filter cake is washed with cold methanol, dried at 50 ℃ under vacuum, to get yellow solid 165.0 mg, yield 79.86%.

[0269] 1H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.42 (s, 1H), 8.00 (s, 1H), 7.95 - 7.89 (m, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.55 Hz, 1H), 7.71 (d, J = 7.93 Hz, 1H), 7.64 (d, J = 7.61 Hz, 1H), 7.59 (t, J = 7.62 Hz, 1H), 7.40 (dd, J = 8.14, 16.45 Hz, 3H), 7.29 (t, J = 7.56 Hz, 1H), 6.83 (s, 1H), 6.71 (s, 1H), 6.63 (d, J = 7.68 Hz, 1H), 5.37 (s, 2H), 3.01 (dt, J = 6.98, 14.10 Hz, 1H), 1.18 (d, J = 7.02 Hz, 6H). MS (ESI): m / z 614.60 [M+H] + Mp: 177-178 °C.

[0270] Example 28. Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-(2,5-difluorobenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-28)

[0271]

[0272] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2, 5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), 2,5-difluorobenzyl bromide (449.56 mg, 2.17 mmol) in DMF (6 ml) in turn, continue to stir the reaction at room temperature for 2 h. The reaction liquid is dropped into 4 ℃ cold water, solid is precipitated, suction filtration, filter cake is washed with water, 50 ℃ vacuum drying, methanol is beaten, placed in-30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, white solid 107.0 mg, yield 24.49%.

[0273] 2) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(2,5- difluorobenzyl)imidazol-4-yl)methyl)piperazine-2,5-dione (150 mg, 0.37 mmol), 3-p- fluorobenzoylbenzaldehyde (102.08 mg, 0.45 mmol), cesium carbonate (182.17 mg, 0.56 mmol), anhydrous sodium sulfate (105.89 mg, 0.75 mmol), DMF (4 ml), exhaust, nitrogen protection, put into 45 °C oil bath, stir for 14 h. Monitor the reaction by LC-MS, and the starting material point disappears. When the reaction is completed, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtered, the filter cake is washed with cold water, the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, and concentrated under reduced pressure. Ultrasonic slurry with methanol, placed in -30 °C refrigerator, suction filtered, the filter cake is washed with cold methanol, and dried at 50 °C under vacuum to obtain yellow solid 79.1 mg, yield 37.19%.

[0274] 1 H NMR (600 MHz, DMSO-d6) δ 11.95 (s, 1H), 10.38 (s, 1H), 8.02 (s, 1H), 7.91 (dd, J = 5.60, 8.53 Hz, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.60 Hz, 1H), 7.64 (d, J = 7.66 Hz, 1H), 7.59 (t, J = 7.64 Hz, 1H), 7.40 (t, J = 8.74 Hz, 2H), 7.34 (td, J = 4.42, 9.29 Hz, 1H), 7.26 (ddd, J = 3.35, 8.13, 11.87 Hz, 1H), 6.91 (ddd, J = 3.28, 5.59, 8.62 Hz, 1H), 6.82 (s, 1H), 6.68 (s, 1H), 5.39 (s, 2H), 3.20 - 3.09 (m, 1H), 1.18 (d, J = 7.08 Hz, 6H). MS (ESI): m / z 571.20 [M+H] + Mp: 193-195 °C.

[0275] Example 29 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylidene-6-((5-isopropyl-1-(2,6-difluorobenzyl)imidazol-4-yl)methylidene)piperazine-2,5-dione (PLN-3-29)

[0276]

[0277] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H- imidazol-4-yl)methylene)piperazine-2,5-dione (200 mg, 0.72 mmol), potassium carbonate (300.20 mg, 2.17 mmol) successively, drop 2,6-difluorobenzyl bromide (299.8 mg, 1.45 mmol) in DMF (6 ml), stir the reaction at 45 °C for 2 h. Drop the reaction into 4 °C cold water, solid precipitates, suction filtration, filter cake water wash, 50 °C vacuum drying, methanol slurry, placed in -30 °C refrigerator for more than 2 h, suction filtration, cold methanol wash, dry, get white solid 174.76 mg, yield 59.99%.

[0278] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1- (2,6-difluorobenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (174.76 mg, 0.43 mmol), 3-p-fluorobenzoyl benzaldehyde (118.95 mg, 0.52 mmol), cesium carbonate (212.27 mg, 0.65 mmol), anhydrous sodium sulfate (123.39 mg, 0.87 mmol), DMF (4 ml), exhaust, nitrogen protection, placed in 45 °C oil bath, stir the reaction for 20 h. LC-MS monitoring reaction, raw material point disappears. Reaction is completed, drop the reaction into 4 °C cold water (40 ml), suction filtration, filter cake cold water wash, the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filter, reduced pressure concentration. Methanol ultrasonic slurry, placed in -30 °C refrigerator, suction filtration, filter cake with cold methanol wash, 50 °C vacuum drying, get yellow solid 191.6 mg, yield 64.43%.

[0279] 1 H NMR (600 MHz, DMSO-d6) δ 11.92 (s, 1H), 10.36 (s, 1H), 7.94-7.88 (m, 3H), 7.81 (s, 1H), 7.75 (d, J = 7.73 Hz, 1H), 7.63 (d, J = 7.70 Hz, 1H), 7.58 (t, J = 7.66 Hz, 1H), 7.54-7.47 (m, 1H), 7.39 (t, J = 8.78 Hz, 2H), 7.18 (t, J = 8.23 Hz, 2H), 6.81 (s, 1H), 6.66 (s, 1H), 5.39 (s, 2H), 3.20 (dt, J = 7.42, 14.87 Hz, 1H), 1.17 (d, J = 7.08 Hz, 6H). MS (ESI): m / z 571.31 [M+H] + Mp: 226-228 °C.

[0280] Example 30: Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((5-isopropyl-1-(2,4-difluorobenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-30)

[0281]

[0282] 1) Take a 25 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (200 mg, 0.72 mmol), potassium carbonate (300.2 mg, 2.17 mmol), anhydrous sodium sulfate (205.7 mg, 0.14 mmol), and dropwise add 6 ml of DMF containing 2,4-difluorobenzyl bromide (299.8 mg, 1.45 mmol). Stir the reaction mixture at room temperature for 2 h. Add the reaction mixture dropwise to 4 °C cold water, and a solid precipitates. Filter the mixture, wash the filter cake with water, dry it under vacuum at 50 °C, slurry it with methanol, and let it stand in a -30 °C refrigerator for at least 2 h. Filter the mixture, wash it with cold methanol, and dry it to obtain 216.2 mg of an off-white solid, with a yield of 74.22%.

[0283] 2) Take a 10ml dry brown round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1-(2,4-difluorobenzylimidazol-4-yl) in sequence. Methylenepiperazine-2,5-dione (150 mg, 0.37 mmol), 3-p-fluorobenzoylbenzaldehyde (102.08 mg, 0.45 mmol), cesium carbonate (182.17 mg, 0.56 mmol), anhydrous sodium sulfate (105.89 mg, 0.75 mmol), and DMF (4 ml) were added. The mixture was vented, protected with nitrogen, and placed in an oil bath at 45 °C with stirring for 19 h. The reaction was monitored by LC-MS until the starting material spot disappeared. After the reaction was complete, the reaction solution was added dropwise to 40 ml of cold water at 4 °C, filtered, and the filter cake was washed with cold water. The filter cake was dissolved in a 1:3 mixture of methanol and dichloromethane, filtered, and concentrated under reduced pressure. The methanol was ultrasonically beaten, placed in a -30 °C refrigerator to stand, filtered, and the filter cake was washed with cold methanol and dried under vacuum at 50 °C to give 175.2 mg of a yellow solid, with a yield of 82.37%.

[0284] 1H NMR (600 MHz, DMSO-d6) δ 11.96 (s, 1H), 10.38 (s, 1H), 8.00 (s, 1H), 7.93-7.89 (m, 2H), 7.82 (s, 1H), 7.76 (d, J = 7.72 Hz, 1H), 7.64 (d, J = 7.71 Hz, 1H), 7.59 (t, J = 7.65 Hz, 1H), 7.43-7.37 (m, 2H), 7.33 (dd, J = 5.69, 15.04 Hz, 1H), 7.12 (dd, J = 4.24, 10.64 Hz, 2H), 6.82 (s, 1H), 6.69 (s, 1H), 5.37 (s, 2H), 3.21-3.10 (m, 1H), 1.17 (d, J = 7.13 Hz, 6H). MS (ESI): m / z 571.04 [M+H] + Mp: 184-188 °C.

[0285] Example 31. Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-(3,4-difluorobenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-31)

[0286]

[0287] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2, 5-dione (200 mg, 0.72 mmol), potassium carbonate (300.2 mg, 2.17 mmol), anhydrous sodium sulfate (205.7 mg, 0.14 mmol), drop 3,4-difluorobenzyl bromide (299.8 mg, 1.45 mmol) in DMF (5 ml), stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 °C cold water, a solid is precipitated, suction filtration, filter cake water washing, 50 °C vacuum drying, methanol slurry, placed in -30 °C refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get white solid 216.9 mg, yield 74.46%.

[0288] 2) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl- 1 -(3,4-difluorobenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.37 mmol), 3-p-fluorobenzoylbenzaldehyde (102.08 mg, 0.45 mmol), cesium carbonate (182.17 mg, 0.56 mmol), anhydrous sodium sulfate (105.89 mg, 0.75 mmol), DMF (4 ml), exhaust, nitrogen protection, put into 45 °C oil bath, stir for 19 h. Monitor the reaction by LC-MS, and the raw material point disappears. When the reaction is completed, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtered, the filter cake is washed with cold water, the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, and concentrated under reduced pressure. Ultrasonic slurry with methanol, placed in -30 °C refrigerator, suction filtered, the filter cake is washed with cold methanol, and vacuum dried at 50 °C to obtain yellow solid 166.1 mg, yield 78.09%.

[0289] 1 H NMR (600 MHz, DMSO-d6) δ 11.98 (s, 1H), 10.38 (s, 1H), 8.07 (s, 1H), 7.94-7.89 (m, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.71 Hz, 1H), 7.64 (d, J = 7.71 Hz, 1H), 7.59 (t, J = 7.65 Hz, 1H), 7.45 (dt, J = 8.52, 10.55 Hz, 1H), 7.42-7.37 (m, 2H), 7.31 (ddd, J = 1.90, 7.69, 10.12 Hz, 1H), 6.97 (d, J = 8.37 Hz, 1H), 6.82 (s, 1H), 6.68 (s, 1H), 5.34 (s, 2H), 3.20-3.04 (m, 1H), 1.14 (d, J = 7.12 Hz, 6H). MS (ESI): m / z 571.03 [M+H] + Mp: 202-204 °C.

[0290] Example 32 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl- 1 -(3,5-difluorobenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-32)

[0291]

[0292] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H- imidazol-4-yl)methylene)piperazine-2,5-dione (200 mg, 0.72 mmol), potassium carbonate (300.2 mg, 2.17 mmol), anhydrous sodium sulfate (205.7 mg, 0.15 mmol), drop 3,5-difluorobenzyl bromide (299.8 mg, 1.45 mmol) in DMF (5 ml), stir the reaction at room temperature for 2 h. Drop the reaction into 4 °C cold water, solid precipitates, suction filtration, filter cake water wash, 50 °C vacuum drying, methanol slurry, placed in -30 °C refrigerator for more than 2 h, suction filtration, cold methanol wash, dry, get white solid 136.9 mg, yield 47.0%.

[0293] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1- (3,5-difluorobenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (100 mg, 0.25 mmol), 3-p-fluorobenzoyl benzaldehyde (68.06 mg, 0.30 mmol), cesium carbonate (121.47 mg, 0.37 mmol), anhydrous sodium sulfate (70.60 mg, 0.50 mmol), DMF (4 ml), exhaust, nitrogen protection, placed in 45 °C oil bath, stir the reaction for 13 h. LC-MS monitoring reaction, raw material point disappears. Reaction, drop the reaction into 4 °C cold water (40 ml), suction filtration, filter cake cold water wash, the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filter, reduced pressure concentration. Methanol ultrasonic slurry, placed in -30 °C refrigerator, suction filtration, filter cake with cold methanol wash, 50 °C vacuum drying, get yellow solid 107.3 mg, yield 50.44%.

[0294] 1 H NMR (600 MHz, DMSO-d6) δ 11.97 (s, 1H), 10.39 (s, 1H), 8.08 (s, 1H), 7.95-7.88 (m, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.72 Hz, 1H), 7.64 (d, J = 7.71 Hz, 1H), 7.59 (t, J = 7.65 Hz, 1H), 7.40 (t, J = 8.79 Hz, 2H), 7.21 (ddd, J = 2.17, 5.74, 9.29 Hz, 1H), 6.88 (d, J = 6.22 Hz, 2H), 6.83 (s, 1H), 6.68 (s, 1H), 5.38 (s, 2H), 3.12 (dt, J = 7.10, 14.25 Hz, 1H), 1.15 (d, J = 7.12 Hz, 6H). MS (ESI): m / z 571.01 [M+H] +Mp: 237-239 °C.

[0295] Example 33 Preparation of (3Z, 6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-(3,4-dichlorobenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-33)

[0296]

[0297] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2,5-dione (200 mg, 0.72 mmol), potassium carbonate (300.2 mg, 2.17 mmol), anhydrous sodium sulfate (205.7 mg, 0.14 mmol), drop 3,4-dichlorobenzyl bromide (347.40 mg, 1.45 mmol) in DMF (5 ml), stir the reaction at room temperature for 2 h. Drop the reaction into 4 °C cold water, solid precipitate, suction filtration, filter cake water wash, methanol and dichloromethane redissolve, reduce pressure concentration, dry, methanol ultrasonic beating, placed in -30 °C refrigerator, suction filtration, filter cake cold methanol wash, 50 °C vacuum drying, get white solid 190.2 mg, yield 60.36%.

[0298] 2) Take 25 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1-(3,4- dichlorobenzyl imidazol-4-yl)methylene)piperazine-2,5-dione (130 mg, 0.30 mmol), 3-p- fluorobenzoyl benzaldehyde (81.79 mg, 0.36 mmol), cesium carbonate (145.97 mg, 0.45 mmol), anhydrous sodium sulfate (84.84 mg, 0.60 mmol), DMF (4 ml), exhaust, nitrogen protection, placed in 45 °C oil bath, stir the reaction for 14 h. LC-MS monitoring reaction, raw material point disappears. Reaction, drop the reaction into 4 °C cold water (40 ml), suction filtration, filter cake cold water wash, filter cake dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduce pressure concentration. Methanol ultrasonic beating, placed in -30 °C refrigerator, suction filtration, filter cake with cold methanol wash, 50 °C vacuum drying, get yellow solid 151.4 mg, yield 84.02%.

[0299] 1H NMR (600 MHz, DMSO-d6) δ 11.97 (s, 1H), 10.38 (s, 1H), 8.08 (s, 1H), 7.93-7.89 (m, 2H), 7.82 (s, 1H), 7.75 (d, J = 7.73 Hz, 1H), 7.64 (dd, J = 5.23, 8.00 Hz, 2H), 7.58 (t, J = 7.66 Hz, 1H), 7.47 (d, J = 1.94 Hz, 1H), 7.42-7.37 (m, 2H), 7.07 (dd, J = 1.95, 8.37 Hz, 1H), 6.82 (s, 1H), 6.67 (s, 1H), 5.36 (s, 2H), 3.11 (1H), 1.14 (d, J = 7.09 Hz, 6H). MS (ESI): m / z 603.05 [M+H] + Mp: 205-207 °C.

[0300] Example 34. Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-(2,6-dichlorobenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-34)

[0301]

[0302] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2, 5-dione (200 mg, 0.72 mmol), potassium carbonate (300.2 mg, 2.17 mmol), anhydrous sodium sulfate (205.7 mg, 0.14 mmol), drop 2,6-dichlorobenzyl bromide (347.40 mg, 1.45 mmol) in DMF (5 ml), stir the reaction under 45 °C oil bath for 2 h. Drop the reaction into 4 °C cold water, solid precipitates, suction filtration, filter cake water wash, methanol and dichloromethane redissolve, reduce pressure concentration, dry, methanol ultrasonic beating, placed in -30 °C refrigerator, suction filtration, filter cake cold methanol wash, 50 °C vacuum drying, white solid 208.5 mg, yield 66.17%.

[0303] 2) Take 25 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(2,6- dichlorobenzyl)imidazol-4-yl)methyl)piperazine-2,5-dione (150 mg, 0.34 mmol), 3-(p- fluorobenzoyl)benzaldehyde (94.37 mg, 0.41 mmol), cesium carbonate (168.42 mg, 0.52 mmol), anhydrous sodium sulfate (97.89 mg, 0.69 mmol), DMF (4 ml), evacuate, nitrogen protection, put into 45 °C oil bath, stir for 12 h. Monitor the reaction by LC-MS, the raw material point disappears. When the reaction is completed, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtration, the filter cake is washed with cold water, the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure. Ultrasonic slurry in methanol, placed in -30 °C refrigerator, suction filtration, the filter cake is washed with cold methanol, vacuum dried at 50 °C, to get 173.0 mg of yellow solid, yield 83.19%.

[0304] 1 H NMR (600 MHz, DMSO-d6) δ 11.89 (s, 1H), 10.37 (s, 1H), 7.94 - 7.89 (m, 2H), 7.82 (s, 1H), 7.75 (d, J = 7.72 Hz, 1H), 7.63 (t, J = 8.17 Hz, 3H), 7.58 (t, J = 7.65 Hz, 1H), 7.52 (dd, J = 7.72, 8.54 Hz, 1H), 7.40 (dd, J = 4.96, 11.87 Hz, 3H), 6.80 (s, 1H), 6.70 (s, 1H), 5.43 (s, 2H), 3.39 - 3.33 (m, 1H), 1.32 (d, J = 7.11 Hz, 6H). MS (ESI): m / z 603.04 [M+H] + Mp: 218-221 °C.

[0305] Example 35 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylidene-6-((5-isopropyl-1-p- trifluoromethylbenzyl)imidazol-4-yl)methyl)piperazine-2,5-dione (PLN-3-35)

[0306]

[0307] 1) Take 25 ml dry round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 H- imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), 4-trifluoromethylbenzyl bromide (519.08 mg, 2.17 mmol) in DMF (6 ml) successively, continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, a solid is precipitated, suction filtration, filter cake water washing, 50 ℃ vacuum drying, methanol slurry, placed in -30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get white solid 300.7 mg, yield 63.75%.

[0308] 2) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(4- trifluoromethyl)benzyl imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.34 mmol), 3- p-fluorobenzoyl benzaldehyde (94.55 mg, 0.41 mmol), potassium carbonate (71.57 mg, 0.52 mmol), anhydrous sodium sulfate (98.09 mg, 0.69 mmol), DMF (3 ml), exhaust, nitrogen protection, placed in 45 ℃ oil bath, stirring reaction 20 h. LC-MS monitoring reaction, raw material point disappears. Reaction, the reaction solution is dropped into 4 ℃ cold water (40 ml), suction filtration, filter cake cold water washing, the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Methanol ultrasonic slurry, placed in -30 ℃ refrigerator, suction filtration, filter cake with cold methanol washing, 50 ℃ vacuum drying, get yellow solid 170.6 mg, yield 81.99%.

[0309] 1 H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.38 (s, 1H), 8.10 (s, 1H), 7.94-7.88 (m, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.72 Hz, 3H), 7.64 (d, J = 7.59 Hz, 1H), 7.59 (t, J = 7.57 Hz, 1H), 7.40 (t, J = 8.44 Hz, 2H), 7.32 (d, J = 7.85 Hz, 2H), 6.83 (s, 1H), 6.69 (s, 1H), 5.48 (s, 2H), 3.09 (dt, J = 7.05, 14.09 Hz, 1H), 1.13 (d, J = 6.95 Hz, 6H). MS (ESI): m / z 603.44 [M+H] + Mp: 216-218 °C.

[0310] Example 36 Preparation of (3Z, 6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-m-trifluoromethylbenzylimidazol-4-yl)methylene)piperazine-2, 5-dione (PLN-3-36)

[0311]

[0312] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2, 5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), 3-trifluoromethylbenzyl bromide (519.08 mg, 2.17 mmol) in DMF (6 ml) in turn, continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, solid is precipitated, suction filtration, filter cake is washed with water, 50 ℃ vacuum drying, methanol is pulsed, placed in-30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get white solid 355.9 mg, yield 75.45%.

[0313] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1-(3-trifluoromethyl)benzylimidazol-4-yl)methylene)piperazine-2, 5-dione (150 mg, 0.34 mmol), 3-p-fluorobenzoyl benzaldehyde (94.55 mg, 0.41 mmol), potassium carbonate (71.57 mg, 0.52 mmol), anhydrous sodium sulfate (98.09 mg, 0.69 mmol), DMF (3 ml) in turn, exhaust, nitrogen protection, placed in 45 ℃ oil bath, stirring reaction 19 h. LC-MS monitoring reaction, raw material point disappears. Reaction is finished, the reaction solution is dropped into 4 ℃ cold water (40 ml), suction filtration, filter cake is washed with cold water, the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Methanol ultrasonic pulsed, placed in-30 ℃ refrigerator, suction filtration, filter cake is washed with cold methanol, 50 ℃ vacuum drying, get yellow solid 137.2 mg, yield 65.94%.

[0314] 1H NMR (500 MHz, DMSO-d6) δ 11.98 (s, 1H), 10.38 (s, 1H), 8.11 (s, 1H), 7.94 - 7.88 (m, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.52 Hz, 1H), 7.69 (d, J = 7.73 Hz, 1H), 7.61 (dt, J = 7.57, 19.56 Hz, 3H), 7.54 (s, 1H), 7.40 (t, J = 8.54 Hz, 3H), 6.82 (s, 1H), 6.68 (s, 1H), 5.47 (s, 2H), 3.13 (dt, J = 6.97, 14.04 Hz, 1H), 1.12 (d, J = 6.99 Hz, 6H). MS (ESI): m / z 603.00 [M+H] + Mp: 203-206 °C.

[0315] Example 37. Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-o-trifluoromethylbenzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-37)

[0316]

[0317] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2, 5-dione (200 mg, 0.72 mmol), potassium carbonate (300.13 mg, 2.17 mmol) in turn, 2-trifluoromethylbenzyl bromide (346.04 mg, 1.45 mmol) of DMF (4 ml) was added under stirring at room temperature, continue to stir the reaction at room temperature for 2 h. The reaction liquid was dropped into 4 ℃ cold water, solid was precipitated, suction filtration, filter cake was washed with water, 50 ℃ vacuum drying, methanol was pulsed, placed in-30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, white solid 215.3 mg, yield 68.46%.

[0318] 2) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(2- trifluoromethyl)benzyl imidazol-4-yl)methylene)piperazine-2,5-dione (120 mg, 0.28 mmol), 3- p-fluorobenzoylbenzaldehyde (75.04 mg, 0.33 mmol), potassium carbonate (57.26 mg, 0.41 mmol), anhydrous sodium sulfate (78.47 mg, 0.55 mmol), DMF (3 ml), evacuate, nitrogen protection, put into 45 °C oil bath, stir the reaction for 19 h. LC-MS monitor the reaction, the raw material point disappears. When the reaction is completed, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtration, the filter cake is washed with cold water, the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure. Methanol ultrasonic slurry, placed in -30 °C refrigerator, suction filtration, the filter cake is washed with cold methanol, vacuum dried at 50 °C, to get yellow solid 85.3 mg, yield 51.25%.

[0319] 1 H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.40 (s, 1H), 8.05 (s, 1H), 7.95 - 7.89 (m, 2H), 7.83 (s, 2H), 7.76 (d, J = 7.50 Hz, 1H), 7.66 (dd, J = 7.82, 16.13 Hz, 2H), 7.57 (dt, J = 7.52, 20.10 Hz, 2H), 7.40 (t, J = 8.51 Hz, 2H), 6.83 (s, 1H), 6.72 (s, 1H), 6.63 (d, J = 7.83 Hz, 1H), 5.53 (s, 2H), 2.92 (dt, J = 6.91, 14.13 Hz, 1H), 1.16 (d, J = 6.98 Hz, 6H). MS (ESI): m / z 603.23 [M+H] + Mp: 227-229 °C.

[0320] Example 38 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-p- trifluoromethoxybenzyl imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-38)

[0321]

[0322] 1) Take 25 ml dry round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 H- imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), 4-trifluoromethoxybenzyl bromide (553.82 mg, 2.17 mmol) in DMF (6 ml) successively, continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, a solid is precipitated, suction filtration, filter cake water washing, 50 ℃ vacuum drying, methanol slurry, placed in -30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get white solid 286.2 mg, yield 58.52%.

[0323] 2) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(4- trifluoromethoxybenzyl imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.33 mmol), 3- p-fluorobenzoyl benzaldehyde (91.20 mg, 0.40 mmol), potassium carbonate (69.03 mg, 0.50 mmol), anhydrous sodium sulfate (94.61 mg, 0.67 mmol), DMF (3 ml), exhaust, nitrogen protection, placed in 45 ℃ oil bath, stir for 15 h. LC-MS monitoring reaction, raw material point disappears. Reaction, the reaction solution is dropped into 4 ℃ cold water (40 ml), suction filtration, filter cake cold water washing, the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Methanol ultrasonic slurry, placed in -30 ℃ refrigerator, suction filtration, filter cake with cold methanol washing, 50 ℃ vacuum drying, get yellow solid 103.0 mg, yield 50.00%.

[0324] 1 H NMR (500 MHz, DMSO-d6) δ 12.00 (s, 1H), 10.41 (s, 1H), 8.09 (s, 1H), 7.95-7.88 (m, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.57 Hz, 1H), 7.64 (d, J = 7.60 Hz, 1H), 7.58 (t, J = 7.62 Hz, 1H), 7.43-7.37 (m, 4H), 7.26 (d, J = 8.31 Hz, 2H), 6.82 (s, 1H), 6.67 (s, 1H), 5.40 (s, 2H), 3.11 (dt, J = 7.00, 14.18 Hz, 1H), 1.12 (d, J = 7.03 Hz, 6H). MS (ESI): m / z 619.13 [M+H] + Mp: 198-200 ℃.

[0325] Example 39 Preparation of (3Z, 6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-o-trifluoromethoxybenzylimidazol-4-yl)methylene)piperazine-2, 5-dione (PLN-3-39)

[0326]

[0327] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2, 5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), 4-trifluoromethoxybenzyl bromide (553.82 mg, 2.17 mmol) in DMF (6 ml) successively, continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, a solid is precipitated, suction filtration, filter cake water washing, 50 ℃ vacuum drying, methanol slurry, placed in -30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get white solid 204.8 mg, yield 41.88%.

[0328] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1-(3-trifluoromethoxybenzyl imidazol-4-yl)methylene)piperazine-2, 5-dione (150 mg, 0.33 mmol), 3-p-fluorobenzoyl benzaldehyde (91.20 mg, 0.40 mmol), potassium carbonate (69.03 mg, 0.50 mmol), anhydrous sodium sulfate (94.61 mg, 0.67 mmol), DMF (3 ml) successively, exhaust, nitrogen protection, placed in 45 ℃ oil bath, stirring reaction for 17 h. LC-MS monitoring reaction, raw material point disappears. Reaction, the reaction solution is dropped into 4 ℃ cold water (40 ml), suction filtration, filter cake cold water washing, the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Methanol ultrasonic slurry, placed in -30 ℃ refrigerator, suction filtration, filter cake with cold methanol washing, 50 ℃ vacuum drying, get yellow solid 156.7 mg, yield 76.07%.

[0329] 1H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.42 (s, 1H), 8.11 (s, 1H), 7.95 - 7.89 (m, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.56 Hz, 1H), 7.64 (d, J = 7.67 Hz, 1H), 7.58 (t, J = 7.62 Hz, 1H), 7.52 (t, J = 8.16 Hz, 1H), 7.40 (t, J = 8.71 Hz, 2H), 7.32 (d, J = 8.19 Hz, 1H), 7.15 (d, J = 5.82 Hz, 2H), 6.82 (s, 1H), 6.67 (s, 1H), 5.42 (s, 2H), 3.12 (dt, J = 6.99, 14.12 Hz, 1H), 1.11 (d, J = 7.05 Hz, 6H). MS (ESI): m / z 619.49 [M+H] + Mp: 208-210 °C.

[0330] Example 40. Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-p-cyanobenzyl imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-40)

[0331]

[0332] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2, 5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), 4-cyanobenzyl bromide (425.72 mg, 2.17 mmol) in DMF (6 ml) in turn, continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 °C cold water, solid is precipitated, suction filtration, filter cake is washed with water, 50 °C vacuum drying, methanol is pulsed, placed in -30 °C refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, white solid 351.4 mg, yield 82.68%.

[0333] 2) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(4- cyano)benzyl imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.38 mmol), 3- p-fluorobenzoylbenzaldehyde (104.96 mg, 0.46 mmol), cesium carbonate (187.28 mg, 0.57 mmol), anhydrous sodium sulfate (108.86 mg, 0.77 mmol), DMF (3.5 ml), evacuate, nitrogen protection, put into 45 °C oil bath, stir for 20 h. Monitor the reaction by LC-MS, the raw material point disappears. When the reaction is completed, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtration, the filter cake is washed with cold water, the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure, and dried. Methanol is ultrasonically beaten, placed in a-30 °C refrigerator, suction filtered, the filter cake is washed with cold methanol. Chromatography, reduced pressure concentration, drying, methanol ultrasonic beating, placed in a-30 °C refrigerator, suction filtered, the filter cake is washed with cold methanol, vacuum dried at 50 °C, to get 89.5 mg of yellow solid, yield 41.74%.

[0334] 1 H NMR (500 MHz, DMSO-d6) δ 11.98 (s, 1H), 10.42 (s, 1H), 8.10 (s, 1H), 7.94 - 7.81 (m, 5H), 7.76 (d, J = 7.43 Hz, 1H), 7.64 (d, J = 7.51 Hz, 1H), 7.59 (t, J = 7.58 Hz, 1H), 7.40 (t, J = 8.49 Hz, 2H), 7.29 (d, J = 7.90 Hz, 2H), 6.83 (s, 1H), 6.68 (s, 1H), 5.48 (s, 2H), 3.06 (dt, J = 6.96, 14.05 Hz, 1H), 1.12 (d, J = 6.94 Hz, 6H). MS (ESI): m / z 560.28 [M+H] + Mp: 162-164 °C.

[0335] Example 41 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-m- cyanobenzyl imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-41)

[0336]

[0337] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H- imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), 3-cyanobenzyl bromide (425.72 mg, 2.17 mmol) in DMF (6 ml) successively, continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, a solid is precipitated, suction filtration, filter cake water washing, 50 ℃ vacuum drying, methanol slurry, placed in -30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get white solid 294.1 mg, yield 69.20%.

[0338] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1- (3-cyanobenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.38 mmol), 3- fluorobenzoylbenzaldehyde (104.96 mg, 0.46 mmol), potassium carbonate (79.44 mg, 0.57 mmol), anhydrous sodium sulfate (108.86 mg, 0.77 mmol), DMF (3.5 ml), exhaust, nitrogen protection, placed in 45 ℃ oil bath, stirring reaction for 20 h. LC-MS monitoring reaction, raw material point disappears. After the reaction, the reaction solution is dropped into 4 ℃ cold water (40 ml), suction filtration, filter cake cold water washing, the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration, drying. Methanol ultrasonic slurry, placed in -30 ℃ refrigerator, suction filtration, filter cake with cold methanol washing, 50 ℃ vacuum drying, get yellow solid 178.6 mg, yield 83.29%.

[0339] 1 H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.41 (s, 1H), 8.10 (s, 1H), 7.93-7.88 (m, 2H), 7.85-7.78 (m, 2H), 7.75 (d, J = 7.49 Hz, 1H), 7.68 (s, 1H), 7.61 (dt, J = 7.62, 23.34 Hz, 3H), 7.46-7.37 (m, 3H), 6.82 (s, 1H), 6.67 (s, 1H), 5.42 (s, 2H), 3.11 (dt, J = 7.03, 14.16 Hz, 1H), 1.12 (d, J = 7.00 Hz, 6H). MS (ESI): m / z 560.25 [M+H] + Mp: 209-212 °C.

[0340] Example 42 Preparation of (3Z, 6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-(3,5-dimethylbenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-42)

[0341]

[0342] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2,5-dione (200 mg, 0.72 mmol), potassium carbonate (300.2 mg, 2.17 mmol), anhydrous sodium sulfate (205.7 mg, 0.14 mmol), drop 3,5-dimethylbenzyl bromide (288.28 mg, 1.45 mmol) in DMF (5 ml), stir the reaction at 45 °C for 2 h. Drop the reaction into 4 °C cold water, solid precipitate, suction filtration, filter cake washed with water, redissolved in methanol and dichloromethane, get the crude oil 158.59 mg, yield 55.64%

[0343] 2) Take 25 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1-(3,5- dimethylbenzyl imidazol-4-yl)methylene)piperazine-2,5-dione (158.59 mg, 0.70 mmol), 3-p- fluorobenzoylbenzaldehyde (228.44 mg, 0.58 mmol), cesium carbonate (283.04 mg, 0.87 mmol), anhydrous sodium sulfate (164.52 mg, 1.16 mmol), DMF (4 ml), exhaust, nitrogen protection, placed in 45 °C oil bath, stirred for 16 h. LC-MS monitoring reaction, raw material point disappears. When the reaction is completed, the reaction liquid is dropped into 4 °C cold water (50 ml), suction filtration, filter cake washed with cold water, the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filtered, concentrated under reduced pressure. Methanol ultrasonic slurry, placed in-30 °C refrigerator, suction filtration, filter cake washed with cold methanol, 50 °C vacuum drying, get yellow solid 110.5 mg, yield 28.26%.

[0344] 1H NMR (600 MHz, DMSO-d6) δ 12.01 (s, 1H), 10.36 (s, 1H), 8.03 (s, 1H), 7.94-7.88 (m, 2H), 7.82 (s, 1H), 7.76 (d, J = 7.65 Hz, 1H), 7.63 (d, J = 7.69 Hz, 1H), 7.58 (t, J = 7.65 Hz, 1H), 7.40 (t, J = 8.77 Hz, 2H), 6.92 (s, 1H), 6.81 (s, 1H), 6.72 (s, 2H), 6.68 (s, 1H), 5.25 (s, 2H), 3.10 (tt, J = 7.18, 14.58 Hz, 1H), 2.23 (s, 6H), 1.14 (d, J = 7.11 Hz, 6H). MS (ESI): m / z 563.64 [M+H] + Mp: 167-170 °C.

[0345] Example 43. Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-(3,5-ditrifluoromethylbenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-43)

[0346]

[0347] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2,5-dione (200 mg, 0.72 mmol), potassium carbonate (300.2 mg, 2.17 mmol), anhydrous sodium sulfate (205.7 mg, 0.14 mmol), drop 3,5-ditrifluoromethylbenzyl bromide (444.6 mg, 1.45 mmol) in DMF (5 ml), stir the reaction at 45 °C for 2 h. Drop the reaction into 4 °C cold water, solid precipitates, suction filtration, filter cake water wash, methanol and dichloromethane redissolve, reduce pressure concentration, dry, methanol ultrasonic beating, placed in -30 °C refrigerator, suction filtration, filter cake with cold methanol wash, 50 °C vacuum drying, get white solid 137.2 mg, yield 37.72%

[0348] 2) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(3,5- dinitrobenzyl)imidazol-4-yl)methyl)piperazine-2,5-dione (100 mg, 0.20 mmol), 3-(p- fluorobenzoyl)benzaldehyde (54.50 mg, 0.24 mmol), cesium carbonate (97.29 mg, 0.30 mmol), anhydrous sodium sulfate (56.54 mg, 0.40 mmol), DMF (4 ml), evacuate, nitrogen protection, put into 45 °C oil bath, stir the reaction for 13 h. LC-MS monitor the reaction, the raw material point disappears. When the reaction is completed, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtration, the filter cake is washed with cold water, the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure. Methanol ultrasonic slurry, placed in -30 °C refrigerator, suction filtration, the filter cake is washed with cold methanol, vacuum dried at 50 °C, to get yellow solid 70.6 mg, yield 52.90%.

[0349] 1 H NMR (600 MHz, DMSO-d6) δ 11.94 (s, 1H), 10.39 (s, 1H), 8.14 (s, 1H), 8.10 (s, 1H), 7.93-7.89 (m, 2H), 7.85 (s, 2H), 7.82 (s, 1H), 7.76 (d, J = 7.72 Hz, 1H), 7.64 (d, J = 7.72 Hz, 1H), 7.58 (t, J = 7.66 Hz, 1H), 7.43-7.37 (m, 2H), 6.82 (s, 1H), 6.67 (s, 1H), 5.57 (s, 2H), 3.20-3.11 (m, 1H), 1.12 (d, J = 7.10 Hz, 6H). MS (ESI): m / z 671.51 [M+H] + Mp: 234-236 °C.

[0350] Example 44 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methyl-6-((5-isopropyl-1 -(3,5- dimethoxybenzyl)imidazol-4-yl)methyl)piperazine-2,5-dione (PLN-3-44)

[0351]

[0352] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H- imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.09 mmol), potassium carbonate (450.45 mg, 3.26 mmol), anhydrous sodium sulfate (308.55 mg, 2.17 mmol), drop 3,5-dimethoxybenzyl bromide (501.9 mg, 2.17 mmol) in DMF (5 ml), stir the reaction at room temperature for 2 h. Drop the reaction into 4 °C cold water, solid precipitates, suction filtration, filter cake washed with water, redissolve in methanol and dichloromethane, reduce pressure concentration, dry, methanol ultrasonic beating, placed in -30 °C refrigerator, suction filtration, filter cake washed with cold methanol, 50 °C vacuum drying, get white solid 77.2 mg, yield 16.67%.

[0353] 2) Take 25 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1- (3,5-dimethoxybenzyl)imidazol-4-yl)methylene)piperazine-2,5-dione (200 mg, 0.47 mmol), 3-p-fluorobenzoylbenzaldehyde (128.42 mg, 0.56 mmol), cesium carbonate (229.18 mg, 0.70 mmol), anhydrous sodium sulfate (133.22 mg, 0.94 mmol), DMF (5 ml), exhaust, nitrogen protection, placed in 45 °C oil bath, stir the reaction for 17 h. LC-MS monitoring reaction, raw material point disappears. Reaction, drop the reaction into 4 °C cold water (50 ml), suction filtration, filter cake washed with cold water, dissolve the filter cake in methanol and dichloromethane (1:3) mixed solution, filter, reduce pressure concentration. Methanol ultrasonic beating, placed in -30 °C refrigerator, suction filtration, filter cake washed with cold methanol, 50 °C vacuum drying, get yellow solid 117.80 mg, yield 42.24%.

[0354] 1H NMR (600 MHz, DMSO-d6) δ 12.00 (s, 1H), 10.37 (s, 1H), 8.05 (s, 1H), 7.91 (dd, J = 5.59, 8.55 Hz, 2H), 7.82 (s, 1H), 7.75 (d, J = 7.66 Hz, 1H), 7.64 (d, J = 7.67 Hz, 1H), 7.59 (t, J = 7.65 Hz, 1H), 7.40 (t, J = 8.75 Hz, 2H), 6.82 (s, 1H), 6.69 (s, 1H), 6.44 (s, 1H), 6.27 (d, J = 1.82 Hz, 2H), 5.25 (s, 2H), 3.70 (s, 6H), 3.15 (dt, J = 7.06, 14.19 Hz, 1H), 1.17 (d, J = 7.10 Hz, 6H). MS (ESI): m / z 595.46 [M+H] + Mp: 201-203 °C.

[0355] Example 45. Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-p- phenylbenzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-45)

[0356]

[0357] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2,5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), anhydrous sodium sulfate (308.6 mg, 2.17 mmol), 4-bromomethylbiphenyl (536.85 mg, 2.17 mmol) in DMF (6 ml) in turn, stir the reaction at 45 °C for 2 h. The reaction liquid is dropped into 4 °C cold water, solid is precipitated, suction filtration, filter cake is washed with water, 50 °C vacuum drying, methanol is pulsed, placed in -30 °C refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, white solid 367.1 mg, yield 76.40%.

[0358] 2) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl- 1 -(4-phenylbenzyl)imidazol-4-yl)methyl)piperazine-2,5-dione (150 mg, 0.34 mmol), 3-p-fluorobenzoylbenzaldehyde (92.84 mg, 0.41 mmol), cesium carbonate (165.68 mg, 0.51 mmol), anhydrous sodium sulfate (96.29 mg, 0.68 mmol), DMF (4 ml), evacuate, nitrogen protection, put into 45 °C oil bath, stir for 15 h. Monitor the reaction by LC-MS, the raw material point disappears. When the reaction is completed, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtration, the filter cake is washed with cold water, the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure. Ultrasonic slurry with methanol, placed in-30 °C refrigerator, suction filtration, the filter cake is washed with cold methanol, vacuum dried at 50 °C, to get yellow solid 146.6 mg, yield 70.81%.

[0359] 1 H NMR (500 MHz, DMSO-d6) δ 12.03 (s, 1H), 10.41 (s, 1H), 8.11 (s, 1H), 7.95-7.88 (m, 2H), 7.83 (s, 1H), 7.76 (d, J = 7.46 Hz, 1H), 7.63 (ddd, J = 7.80, 15.24, 29.98 Hz, 6H), 7.49-7.32 (m, 5H), 7.22 (d, J = 7.85 Hz, 2H), 6.82 (s, 1H), 6.69 (s, 1H), 5.39 (s, 2H), 3.17 (dt, J = 6.95, 13.99 Hz, 1H), 1.15 (d, J = 6.95 Hz, 6H). MS (ESI): m / z 611.14 [M+H] + Mp: 160-163 °C.

[0360] Example 46 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylidene-6-((5- isopropyl-1-(2-naphthylmethyl)imidazol-4-yl)methylidene)piperazine-2,5-dione (PLN-3-46)

[0361]

[0362] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl) methylene)piperazine-2,5-dione (300 mg, 1.09 mmol), potassium carbonate (450.16 mg, 3.26 mmol), anhydrous sodium sulfate (308.6 mg, 2.17 mmol), 2-bromomethyl naphthalene (480.15 mg, 2.17 mmol) in DMF (6 ml) in order, stir the reaction at 45 °C for 2 h. Drop the reaction into 4 °C cold water, solid precipitates, suction filtration, filter cake water wash, 50 °C vacuum drying, methanol slurry, put in -30 °C refrigerator for more than 2 h, suction filtration, cold methanol wash, dry, get white solid 236.3 mg, yield 52.25%.

[0363] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1-(2- naphthalenylmethyl imidazol-4-yl) methylene)piperazine-2,5-dione (150 mg, 0.36 mmol), 3- p-fluorobenzoylbenzaldehyde (98.64 mg, 0.43 mmol), cesium carbonate (176.01 mg, 0.54 mmol), anhydrous sodium sulfate (102.31 mg, 0.72 mmol), DMF (4 ml), exhaust, nitrogen protection, put in 45 °C oil bath, stir the reaction for 21 h. LC-MS monitor the reaction, raw material point disappears. Reaction is completed, drop the reaction into 4 °C cold water (40 ml), suction filtration, filter cake cold water wash, filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filter, reduce pressure concentration. Methanol ultrasonic slurry, put in -30 °C refrigerator, suction filtration, filter cake with cold methanol wash, 50 °C vacuum drying, get yellow solid 136.9 mg, yield 65.01%.

[0364] 1 H NMR (500 MHz, DMSO-d6) δ 12.05 (s, 1H), 10.41 (s, 1H), 8.15 (s, 1H), 7.91 (td, J = 6.95, 12.58 Hz, 5H), 7.84 (s, 1H), 7.76 (d, J = 7.56 Hz, 1H), 7.66 - 7.56 (m, 3H), 7.53 - 7.48 (m, 2H), 7.40 (t, J = 8.70 Hz, 2H), 7.31 (d, J = 8.50 Hz, 1H), 6.83 (s, 1H), 6.69 (s, 1H), 5.52 (s, 2H), 3.17 (dt, J = 7.03, 14.20 Hz, 1H), 1.10 (d, J = 7.04 Hz, 6H). MS (ESI): m / z 585.48 [M+H] + Mp: 166-168 °C.

[0365] Example 47 Preparation of (3Z, 6Z)-3-(3-(p-Fluorobenzoyl)phenyl)methylene-6-((5- isopropyl-1-p-toluoylbenzylimidazol-4-yl)methylene)piperazine-2, 5-dione (PLN-3-47)

[0366]

[0367] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2, 5-dione (200 mg, 0.72 mmol), potassium carbonate (300.2 mg, 2.17 mmol) in sequence, drop 4-benzoylbenzyl bromide (398.4 mg, 1.45 mmol) in DMF (6 ml), stir the reaction at room temperature for 2 h. Drop the reaction into 4 °C cold water, solid precipitates, suction filtration, filter cake water wash, 50 °C vacuum drying, get white solid 226.9 mg, yield 66.61%.

[0368] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1-(4- benzoylbenzylimidazol-4-yl)methylene)piperazine-2, 5-dione (150 mg, 0.32 mmol), 3-p- fluorobenzoylbenzaldehyde (87.29 mg, 0.38 mmol), potassium carbonate (66.09 mg, 0.48 mmol), anhydrous sodium sulfate (90.56 mg, 0.64 mmol), DMF (3 ml), exhaust, nitrogen protection, put in 45 °C oil bath, stir the reaction for 30 h. LC-MS monitoring reaction, raw material point disappears. Reaction, drop the reaction into 4 °C cold water (40 ml), suction filtration, filter cake cold water wash, filter cake dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration, drying. Methanol ultrasonic slurry, placed in-30 °C refrigerator, suction filtration, filter cake cold methanol wash. Chromatography, reduced pressure concentration, drying, MeOH slurry, methanol ultrasonic slurry, placed in-30 °C refrigerator, suction filtration, filter cake with cold methanol wash, 50 °C vacuum drying, get yellow solid 105.0 mg, yield 51.57%.

[0369] 1H NMR (500 MHz, DMSO-d6) δ 12.01 (s, 1H), 10.41 (s, 1H), 8.12 (s, 1H), 7.95 - 7.88 (m, 2H), 7.83 (s, 1H), 7.75 (d, J = 7.76 Hz, 3H), 7.72 - 7.63 (m, 4H), 7.57 (dt, J = 7.51, 14.83 Hz, 3H), 7.40 (t, J = 8.34 Hz, 2H), 7.29 (d, J = 7.76 Hz, 2H), 6.82 (s, 1H), 6.69 (s, 1H), 5.49 (s, 2H), 3.12 (dt, J = 6.92, 13.98 Hz, 1H), 1.15 (d, J = 6.96 Hz, 6H). MS (ESI): m / z 639.30 [M+H] + Mp: 175-177 °C.

[0370] Example 48 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-(5-isopropyl-1-(4-nitro)benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-48)

[0371]

[0372] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2,5-dione (200 mg, 0.72 mmol), potassium carbonate (300.13 mg, 2.17 mmol), DMF (4 ml) in turn, stir at room temperature for 20 min, then add p-nitrobenzyl bromide (247.63 mg, 1.45 mmol), continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, solid is precipitated, suction filtration, filter cake is washed with water, 50 ℃ vacuum drying, methanol is pulsed, placed in -30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get brown yellow solid 252 mg, yield 95.00%.

[0373] 2) Take 25 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(4- nitro)benzyl imidazol-4-yl)methylene)piperazine-2,5-dione (200 mg, 0.49 mmol), 3-p- fluorobenzoylbenzaldehyde (133.12 mg, 0.58 mmol), DBN (90.55 mg, 0.73 mmol), anhydrous sodium sulfate (138.09 mg, 0.97 mmol), DMF (4 ml), exhaust, nitrogen protection, put into 45 °C oil bath, stir the reaction for 12 h. Monitor the reaction by LC-MS, and the raw material point disappears. When the reaction is completed, the reaction solution is dropped into 4 °C cold water (40 ml), suction filtered, and the filter cake is washed with cold water (30 ml*2). The filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure, loaded on C18, and purified by reverse column chromatography with gradient elution and concentrated under reduced pressure to obtain 62.3 mg of a light yellow solid, with a yield of 22.11%.

[0374] 1 H NMR (500 MHz, DMSO-d6) δ 11.98 (s, 1H), 10.39 (s, 1H), 8.18 (d, J = 72.15 Hz, 3H), 8.05 - 7.72 (m, 4H), 7.63 (s, 2H), 7.38 (s, 4H), 6.76 (d, J = 68.46 Hz, 2H), 5.54 (s, 2H), 3.07 (s, 1H), 1.14 (s, 6H). MS (ESI): m / z 580.32 [M+H] + Mp: 152-154 °C.

[0375] Example 49 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-(5-isopropyl-1-(3- nitro)benzyl imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-49)

[0376]

[0377] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H- imidazol-4-yl)methyl)piperazine-2,5-dione (500 mg, 1.81 mmol), potassium carbonate (750.26 mg, 5.43 mmol), DMF (10 ml) in turn, stir for 10 min at room temperature, then add m-nitrobenzyl bromide (781.88 mg, 3.62 mmol), continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, a solid is precipitated, suction filtration, filter cake water washing, 50 ℃ vacuum drying, methanol slurry, placed in -30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get white solid 608.1 mg, yield 81.68%.

[0378] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1- (3-nitro) benzyl imidazol-4-yl)methyl)piperazine-2,5-dione (500 mg, 1.22 mmol), 3- p-fluorobenzoyl benzaldehyde (332.84 mg, 1.46 mmol), potassium carbonate (251.94 mg, 1.82 mmol), anhydrous sodium sulfate (345.24 mg, 2.43 mmol), DMF (5 ml), exhaust, nitrogen protection, placed in 45 ℃ oil bath, stir for 19 h. LC-MS monitoring reaction, raw material point disappears. Reaction is finished, the reaction solution is dropped into 4 ℃ cold water (30 ml), suction filtration, filter cake cold water washing (20 ml*2), the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Methanol ultrasonic slurry, placed in -30 ℃ refrigerator, suction filtration, filter cake with cold methanol washing, again with methanol and dichloromethane (10:1) ultrasonic slurry, placed in -30 ℃ refrigerator, suction filtration, filter cake with cold methanol washing, 50 ℃ vacuum drying, get yellow solid 482.3 mg, yield 68.47%.

[0379] 1H NMR (500 MHz, DMSO-d6) δ 11.93 (s, 1H), 10.38 (s, 1H), 8.18 (dd, J = 1.42, 8.15 Hz, 1H), 8.12 (s, 1H), 8.04 (s, 1H), 7.96 - 7.88 (m, 2H), 7.82 (s, 1H), 7.75 (d, J = 7.65 Hz, 1H), 7.69 (t, J = 7.97 Hz, 1H), 7.64 (d, J = 7.72 Hz, 1H), 7.58 (dd, J = 8.09, 16.01 Hz, 2H), 7.40 (t, J = 8.82 Hz, 2H), 6.82 (s, 1H), 6.68 (s, 1H), 5.59 - 5.43 (m, 2H), 3.13 (dt, J = 7.10, 14.24 Hz, 1H), 1.29 - 0.95 (m, 6H). MS (ESI): m / z 580.21 [M+H] + . Mp: 250-252 °C.

[0380] Example 50 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-(5- isopropyl-1-(2-nitro)benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-50)

[0381]

[0382] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2,5-dione (200 mg, 0.72 mmol), potassium carbonate (300.1 mg, 2.17 mmol), DMF (4 ml) in turn, stir for 10 min at room temperature, then add 2- nitrobenzyl bromide (781.88 mg, 3.62 mmol), continue to stir the reaction for 2 h at room temperature. The reaction solution is dropped into 4 °C cold water, solid is precipitated, suction filtration, filter cake is washed with water, 50 °C vacuum drying, methanol is pulsed, placed in -30 °C refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get 225.7 mg of yellow solid, yield 75.78%.

[0383] 2) Take 10 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(2- nitro)benzyl imidazol-4-yl)methylene)piperazine-2,5-dione (100 mg, 0.24 mmol), 3-p- fluorobenzoylbenzaldehyde (66.57 mg, 0.29 mmol), potassium carbonate (50.42 mg, 0.36 mmol), anhydrous sodium sulfate (69.05 mg, 0.49 mmol), DMF (3 ml), exhaust, nitrogen protection, put into 45 °C oil bath, stir for 12 h. Monitor the reaction by LC-MS, and the starting material point disappears. When the reaction is completed, the reaction solution is dropped into 4 °C cold water (30 ml), suction filtration, the filter cake is washed with cold water (20 ml*2), the filter cake is dissolved in a mixture of methanol and dichloromethane (1:3), filtered, concentrated under reduced pressure, dried, methanol ultrasonic beating, placed in-30 °C refrigerator, suction filtration, the filter cake is washed with cold methanol, vacuum dried at 50 °C, to get yellow solid 101.0 mg, yield 71.68%.

[0384] 1 H NMR (500 MHz, DMSO-d6) δ 11.98 (s, 1H), 10.40 (s, 1H), 8.20 (dd, J = 0.84, 8.13 Hz, 1H), 7.99 (d, J = 11.03 Hz, 1H), 7.92 (dd, J = 5.58, 8.71 Hz, 2H), 7.82 (d, J = 13.60 Hz, 1H), 7.75 (dd, J = 7.34, 14.25 Hz, 2H), 7.62 (dt, J = 7.73, 24.43 Hz, 3H), 7.45 - 7.35 (m, 2H), 6.83 (s, 1H), 6.73 (s, 1H), 6.60 (t, J = 11.11 Hz, 1H), 5.74 (s, 2H), 3.16 - 2.93 (m, 1H), 1.36 - 1.02 (m, 6H). MS (ESI): m / z 580.18 [M+H] + Mp: 176-179 °C.

[0385] Example 51 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-(5-isopropyl-1-(4- methoxybenzoyl)benzyl imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-51)

[0386]

[0387] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H- imidazol-4-yl)methyl)piperazine-2,5-dione (500 mg, 1.81 mmol), potassium carbonate (750.27 mg, 5.43 mmol), anhydrous sodium sulfate (514.07 mg, 3.62 mmol), DMF (10 ml) in turn, stir for 10 min at room temperature, then add p-methyl formate benzyl bromide (829.05 mg, 3.62 mmol), continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, a solid is precipitated, suction filtration, filter cake water washing, 50 ℃ vacuum drying, methanol slurry, placed in-30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get white solid 653.8 mg, yield 85.12%.

[0388] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1- (4-methoxy acyl) benzyl imidazol-4-yl)methyl)piperazine-2,5-dione (500 mg, 1.18 mmol), 3-p-fluorobenzoyl benzaldehyde (322.77 mg, 1.41 mmol), cesium carbonate (575.98 mg, 1.77 mmol), anhydrous sodium sulfate (334.80 mg, 2.36 mmol), DMF (10 ml) in turn, exhaust, nitrogen protection, placed in 45 ℃ oil bath, stir for 12 h. LC-MS monitoring reaction, raw material point disappears. Reaction is finished, the reaction solution is dropped into 4 ℃ cold water (100 ml), suction filtration, filter cake cold water washing (100 ml*3), the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Methanol ultrasonic slurry, placed in-30 ℃ refrigerator, suction filtration, filter cake with cold methanol washing, 50 ℃ vacuum drying, get yellow solid 476.5 mg, yield 68.22%.

[0389] 1H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.37 (s, 1H), 8.10 (s, 1H), 7.97 (d, J = 8.21 Hz, 2H), 7.91 (dd, J = 5.62, 8.55 Hz, 2H), 7.82 (s, 1H), 7.76 (d, J = 7.64 Hz, 1H), 7.64 (d, J = 7.67 Hz, 1H), 7.59 (t, J = 7.63 Hz, 1H), 7.40 (t, J = 8.75 Hz, 2H), 7.25 (d, J = 8.19 Hz, 1H), 6.82 (s, 1H), 6.68 (s, 1H), 5.46 (s, 2H), 3.84 (s, 3H), 3.07 (dt, J = 7.14, 14.27 Hz, 1H), 1.11 (d, J = 7.08 Hz, 6H). MS (ESI): m / z 593.19 [M+H] + . Mp:202-204℃.

[0390] Example 52 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-(5-isopropyl-1-(3-methoxybenzoyl)benzylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-52)

[0391]

[0392] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2, 5-dione (500 mg, 1.81 mmol), potassium carbonate (750.27 mg, 5.43 mmol), anhydrous sodium sulfate (514.07 mg, 3.62 mmol), DMF (10 ml) in turn, stir at room temperature for 10 min, then add m-carboxylic acid methyl benzyl bromide (829.05 mg, 3.62 mmol), continue to stir the reaction at room temperature for 2 h. The reaction liquid is dropped into 4 ℃ cold water, a solid is precipitated, suction filtration, filter cake water washing, 50 ℃ vacuum drying, methanol slurry, placed in-30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, get white solid 669.4 mg, yield 87.15%.

[0393] 2) Take 25 ml dry brown round bottom flask, add (Z)-1 -acetyl-3-((5-isopropyl-1 -(3- methoxyacetyl)benzyl imidazol-4-yl)methylene)piperazine-2,5-dione (500 mg, 1.18 mmol), 3- p-fluorobenzoylbenzaldehyde (322.77 mg, 1.41 mmol), cesium carbonate (575.98 mg, 1.77 mmol), anhydrous sodium sulfate (334.80 mg, 2.36 mmol), DMF (10 ml), evacuate, nitrogen protection, put into 45 °C oil bath, stir the reaction for 17 h. LC-MS monitor the reaction, the raw material point disappears. When the reaction is completed, the reaction solution is dropped into 4 °C cold water (100 ml), suction filtration, the filter cake is washed with cold water (100 ml*3), the filter cake is dissolved in a mixture of methanol and dichloromethane (1 :3), filtered, concentrated under reduced pressure. Methanol ultrasonic slurry, placed in -30 °C refrigerator, suction filtration, the filter cake is washed with cold methanol, 50 °C vacuum drying, get yellow solid 464.8 mg, yield 66.55%.

[0394] 1 H NMR (500 MHz, DMSO-d6) d 11.99 (s, 1 H), 10.37 (s, 1 H), 8.10 (s, 1 H), 7.95-7.88 (m, 3H), 7.83 (s, 1 H), 7.76 (d, J = 9.23 Hz, 2H), 7.64 (d, J = 7.64 Hz, 1 H), 7.61 -7.52 (m, 2H), 7.40 (t, J = 8.79 Hz, 3H), 6.82 (s, 1 H), 6.68 (s, 1 H), 5.45 (s, 2H), 3.84 (s, 3H), 3.12 (dt, J = 7.04, 14.18 Hz, 1 H), 1.12 (d, J = 7.06 Hz, 6H). MS (ESI): m / z 593.19 [M+H] + Mp: 219-220 °C.

[0395] Example 53 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-(5-isopropyl-1 -(2- methoxyacetyl)benzyl imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-53)

[0396]

[0397] 1) Take 25 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H- imidazol-4-yl)methyl)piperazine-2,5-dione (300 mg, 1.09 mmol), potassium carbonate (450.15 mg, 3.26 mmol), anhydrous sodium sulfate (308.45 mg, 2.17 mmol), DMF (8 ml), stir for 10 min at room temperature, then add methyl o-benzoate benzyl bromide (497.44 mg, 2.17 mmol), continue to stir the reaction at room temperature for 2 h. The reaction solution is dropped into 4 ℃ cold water, a solid is precipitated, suction filtration, filter cake water washing, 50 ℃ vacuum drying, methanol slurry, placed in -30 ℃ refrigerator for more than 2 h, suction filtration, cold methanol washing, drying, white solid 365.10 mg, yield 79.22%.

[0398] 2) Take 10 ml dry brown round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1- (2-methoxy acyl) benzyl imidazol-4-yl)methyl)piperazine-2,5-dione (200 mg, 0.47 mmol), 3-p-fluorobenzoyl benzaldehyde (129.10 mg, 0.57 mmol), cesium carbonate (230.39 mg, 0.71 mmol), anhydrous sodium sulfate (133.92 mg, 0.94 mmol), DMF (4 ml), exhaust, nitrogen protection, placed in 45 ℃ oil bath, stir for 15 h. LC-MS monitoring reaction, raw material point disappears. Reaction is finished, the reaction solution is dropped into 4 ℃ cold water (40 ml), suction filtration, filter cake cold water washing (50 ml*3), the filter cake is dissolved in methanol and dichloromethane (1:3) mixed solution, filtration, reduced pressure concentration. Methanol ultrasonic slurry, placed in -30 ℃ refrigerator, suction filtration, filter cake with cold methanol washing, 50 ℃ vacuum drying, yellow solid 220.5 mg, yield 78.93%.

[0399] 1H NMR (500 MHz, DMSO-d6) δ 12.01 (s, 1H), 10.38 (s, 1H), 7.99 (d, J = 6.53 Hz, 2H), 7.92 (dd, J = 5.76, 8.27 Hz, 2H), 7.83 (s, 1H), 7.77 (d, J = 7.42 Hz, 1H), 7.64 (d, J = 7.60 Hz, 1H), 7.59 (t, J = 7.64 Hz, 2H), 7.46 (t, J = 7.57 Hz, 1H), 7.41 (t, J = 8.71 Hz, 2H), 6.83 (s, 1H), 6.73 (s, 1H), 6.54 (d, J = 7.83 Hz, 1H), 5.69 (s, 2H), 3.89 (s, 3H), 2.99 (dt, J = 7.02, 14.16 Hz, 1H), 1.17 (d, J = 7.05 Hz, 6H). MS (ESI): m / z 593.14 [M+H] + Mp: 234-236 °C.

[0400] Example 54. Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1- cyclohexylmethylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-54)

[0401]

[0402] A 50 ml dry round bottom flask was charged with (Z)-1-acetyl-3-((5-cyclopropyl-1H- imidazol-4-yl)methylene)piperazine-2,5-dione (200 mg, 0.73 mmol), cesium carbonate (471.65 mg, 1.45 mmol), potassium iodide (120.17 mg, 0.72 mmol), 4A molecular sieves (200 mg), bromomethylcyclohexane (192.28 mg, 1.08 mmol) in DMF (4 ml), evacuated, nitrogen protection, placed in 70 °C oil bath, stirred for 24 h. After the reaction, it was cooled to room temperature, and 3-p-fluorobenzoylbenzaldehyde (132.17 mg, 0.58 mmol) in DMF (2 ml) was added, and the temperature was raised to 45 °C oil bath, and stirred for 20 h. The reaction was monitored by LC-MS, and after the reaction, the reaction solution was dropped into 4 °C cold water (80 ml), suction filtered, the filter cake was washed with cold water, suction dried, dissolved in methanol and dichloromethane (1:3), filtered, dried, chromatographed (MeOH:DCM = 1:40), short column separation (MeOH:DCM = 1:10), reduced pressure concentration, drying, EA ultrasonic beating, adding appropriate amount of PE, suction filtering, drying, 50 °C vacuum drying, to get 42.4 mg of yellow solid, yield 10.83%.

[0403] 1 H NMR (600 MHz, DMSO-d6) δ 12.02 (s, 1H), 10.35 (s, 1H), 7.91 (dd, J = 5.58, 8.61 Hz, 2H), 7.84 (s, 1H), 7.82 (d, J = 7.73 Hz, 1H), 7.75 (d, J = 7.66 Hz, 1H), 7.63 (d, J = 7.68 Hz, 1H), 7.58 (t, J = 7.64 Hz, 1H), 7.40 (t, J = 8.77 Hz, 2H), 6.80 (s, 1H), 6.70 (s, 1H), 3.85 (d, J = 7.49 Hz, 2H), 3.19 (dq, J = 6.99, 14.20 Hz, 1H), 1.71 - 1.56 (m, 4H), 1.49 (d, J = 11.69 Hz, 2H), 1.31 (d, J = 7.07 Hz, 6H), 1.14 (d, J = 9.39 Hz, 2H), 0.96 (dd, J = 10.67, 22.47 Hz, 2H). MS (ESI): m / z 541.23 [M+H] + Mp: 163-165 °C.

[0404] Example 55 Preparation of (3Z,6Z)-3-((-3-(4-fluorophenoxy)phenyl)methylene)-6-((5-isopropyl-1-(3- cyclohexylpropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-55)

[0405] 1) 3-cyclohexylpropyl 4-methylbenzenesulfonate

[0406]

[0407] Take 50 ml dry round bottom flask, 0 ℃, 3-cyclohexylpropan-1-ol (2 g, 14.06 mmol) was dissolved in 10 ml of anhydrous DCM, triethylamine (7 g, 70.30 mmol) was added, nitrogen protection, dropwise addition of p-methylbenzenesulfonyl chloride (4 g, 21.09 mmol) dissolved in 10 ml of anhydrous DCM, transfer to room temperature, stir the reaction for 6 h. Dilute the reaction solution with dichloromethane, purify with water (100 ml*2), dry the organic phase with anhydrous sodium sulfate, filter, reduce pressure concentration, purify by column chromatography, PE:EA = 10:1, get white solid 3.9 g, yield 94.66%.

[0408] 2) (Z)-1-acetyl-3-((1-(3-cyclohexylpropyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0409]

[0410] Into a 50 mL dry round bottom flask, (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), 3-cyclohexylpropyl 4- methylbenzenesulfonate (639 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieves (500 mg), DMF (6 mL), nitrogen protection, placed in 70 °C oil bath, stirred for 24 h. Transfer to a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure, obtain orange oil, then redissolve with a mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:5), suction filtration, wash the filter cake with a mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:5), concentrate under reduced pressure, obtain 400 mg of brown oil mixture, without purification, directly proceed to the next step.

[0411] 3) (3Z,6Z)-3-((-3-(4-fluorophenoxy)phenyl)methylene)-6-((5-isopropyl-1-(3- cyclohexylpropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0412]

[0413] Into a 25 mL dry reaction flask, (Z)-1-acetyl-3-((1-(3-cyclohexylpropyl)-5-isopropyl-1H- imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.75 mmol), 3-(4- fluorophenoxy)benzaldehyde (194 mg, 0.90 mmol), cesium carbonate (366 mg, 1.12 mmol), anhydrous sodium sulfate (213 mg, 1.50 mmol), DMF (6 mL), nitrogen protection, light protection, placed in a 45 °C oil bath, stirred for 20 h. Transfer to a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure, then redissolve with a mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:3), suction filtration, wash the filter cake with a mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:3), concentrate under reduced pressure. Flash purification (methanol: 87%, water: 13%), obtain the product, concentrate under reduced pressure, obtain 88 mg of light yellow solid, with a yield of 14.42%.

[0414] 1H NMR (500 MHz, dmso) δ 12.00 (s, 1H), 10.12 (s, 1H), 7.88 (s, 1H), 7.41 (t, J = 7.9 Hz, 1H), 7.26 (d, J = 7.8 Hz, 1H), 7.23 (t, J = 8.7 Hz, 2H), 7.17 (s, 1H), 7.12 (dd, J = 8.8, 4.4 Hz, 2H), 6.90 (d, J = 8.0 Hz, 1H), 6.72 (s, 1H), 6.70 (s, 1H), 3.98 (t, J = 7.0 Hz, 2H), 3.28 - 3.11 (m, 1H), 1.73 - 1.53 (m, 7H), 1.32 (d, J = 7.0 Hz, 6H), 1.22 - 1.08 (m, 6H), 0.83 (dd, J = 22.1, 10.7 Hz, 2H). MS (ESI): m / z 557.20 [M+H] + Mp: 189-191 °C.

[0415] Example 56 Preparation of (3Z,6Z)-3-(3-(4-fluorophenoxy)phenyl)methylene)-6-((5- isopropyl-1-((cyclohexylmethoxy)ethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5- dione (PLN-3-56)

[0416] 1) 2-cyclohexylmethoxy-1-ethanol

[0417]

[0418] Take 50ml dry round bottom flask, in turn (bromomethyl) cyclohexane (2g, 11.30mmol), add 4mL 25.5M NaOH, ethylene glycol (10ml), exhaust, nitrogen protection, placed in 80°C oil bath, stirring reaction 16h. Cool to room temperature, ethyl acetate (100ml*3) extraction, combined organic phase, anhydrous sodium sulfate drying, filtration, reduced pressure concentration, column chromatography purification (PE:EA=5:1), get orange oil liquid 800mg, yield 44.94%.

[0419] 2) 2-(cyclohexylmethoxy)ethyl 4-methylbenzenesulfonate

[0420]

[0421] Take 50 ml dry round-bottom flask, 0 ℃, 2-cyclohexylmethoxy-1-ethanol (1 g, 6.33 mmol) is dissolved in 10 ml of anhydrous DCM, add triethylamine (3.19 g, 31.65 mmol), exhaust, nitrogen protection, drop 10 ml of anhydrous DCM dissolved p-methyl benzene sulfonyl chloride (1.44 g, 7.60 mmol), transfer to room temperature, stir for 6 h. Purification water (100 ml*2) wash, dry the organic phase with anhydrous sodium sulfate, filter, concentrate under reduced pressure, purify by column chromatography, PE:EA = 10:1, get 1.38 g of yellowish oil liquid, yield 70.12%.

[0422] 3) (Z)-1-acetyl-3-((1-(2-(cyclohexylmethoxy)ethyl)-5-isopropyl-1H-imidazol-4-yl) methylene) piperazine-2,5-dione

[0423]

[0424] Take 50 ml dry round-bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl) methylene) piperazine-2,5-dione (300 mg, 1.08 mmol), 2-(cyclohexylmethoxy) ethyl 4-methyl benzene sulfonate (674 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieve (500 mg), DMF (6 ml), nitrogen protection, placed in 70 ℃ oil bath, stirred for 24 h. Transfer to 100 ml single-neck flask, rinse with ethanol, concentrate under reduced pressure with water to get orange oil, then redissolve with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:5), filter, wash the filter cake with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:5), concentrate under reduced pressure to get 400 mg of brown oil mixture, which is not purified and directly used for the next step.

[0425] 4) (3Z,6Z)-3-(3-(4-fluorophenoxy)phenyl)methylene)-6-((5-isopropyl-1-((cyclohexylmethoxy) ethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0426]

[0427] Take 25 mL dry reaction bottle, (Z)-1 -acetyl-3-((5-isopropyl-1 -(2- cyclohexylmethoxyethyl)-1 H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.72 mmol), 3-(4-fluorophenoxy)benzaldehyde (186 mg, 0.86 mmol), cesium carbonate (351 mg, 1.08 mmol), anhydrous sodium sulfate (204 mg, 1.44 mmol), DMF (6 mL), nitrogen protection, light shielding, placed in 45 °C oil bath for 20 h. Transfer to 100 ml single port bottle, ethanol rinse, concentrate under reduced pressure, then redissolve with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), suction filtration, and then wash the filter cake with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), and concentrate under reduced pressure. Methyl alcohol slurry, placed in -20 °C for 2 h, filtered, dried, and 84 mg of light yellow solid was obtained, with a yield of 13.53%.

[0428] 1 H NMR (500 MHz, dmso) δ 11.97 (s, 1H), 10.13 (s, 1H), 7.82 (s, 1H), 7.48 (d, J = 7.8 Hz, 1H), 7.40 (t, J = 7.9 Hz, 1H), 7.28 (d, J = 7.6 Hz, 1H), 7.22 (d, J = 7.9 Hz, 2H), 7.15 - 7.07 (m, 2H), 6.89 (d, J = 8.1 Hz, 1H), 6.69 (s, 2H), 4.18 (t, J = 4.6 Hz, 2H), 3.60 (t, J = 4.6 Hz, 2H), 3.28 - 3.22 (m, 1H), 3.18 (d, J = 6.3 Hz, 2H), 2.29 (s, 1H), 1.62 (t, J = 12.0 Hz, 4H), 1.31 (d, J = 7.0 Hz, 6H), 1.22 - 1.04 (m, 4H), 0.85 (dd, J = 22.6, 11.4 Hz, 2H). MS (ESI): m / z 573.21 [M+H] + .Mp: 168-170 °C.

[0429] Example 57 Preparation of (3Z,6Z)-3-(3-(4-fluorophenoxy)phenyl)methylene)-6-((5-isopropyl-1-(2-phenoxyethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-57)

[0430] 1,2-Phenoxyethyl-1-ol

[0431]

[0432] Take 50 ml dry round-bottom flask, add bromobenzene (2 g, 12.74 mmol) in turn, potassium carbonate (5.28 g, 38.22 mmol), copper chloride (108 mg, 0.64 mmol), ethylene glycol (16 ml), nitrogen protection, placed in 130 ℃ oil bath, stirring reaction 12 h. Cool to room temperature, adjust pH to 3 with dilute hydrochloric acid, extract with ethyl acetate (100 ml*2), combine the organic phase, dry over anhydrous sodium sulfate, filter, reduce pressure and concentrate, purify by column chromatography, PE:EA = 5:1, get 1.1 g of yellowish oil, the yield is 62.50%.

[0433] 2, 2-Phenoxyethyl-4-methylbenzenesulfonate

[0434]

[0435] Take 50 ml dry round-bottom flask, 0 ℃, 2-phenoxyethyl-1-alcohol (1 g, 7.25 mmol) is dissolved in 10 ml of anhydrous DCM, add triethylamine (3.66 g, 36.25 mmol), nitrogen protection, dropwise addition of p-methylbenzenesulfonyl chloride (2.48 g, 13.05 mmol) dissolved in 10 ml of anhydrous DCM, transfer to room temperature, stirring reaction 6 h. Purification with water (100 ml*2), combine the organic phase, dry over anhydrous sodium sulfate, filter, reduce pressure and concentrate, purify by column chromatography, PE:EA = 4:1, get 1.65 g of white solid, the yield is 77.94%.

[0436] 3, (Z)-1-Acetyl-3-((5-isopropyl-1-(2-phenoxyethyl)-1H-imidazol-4-yl) methylene) piperazine-2, 5-dione

[0437]

[0438] Take 50 ml dry round-bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl) methylene) piperazine-2, 5-dione (300 mg, 1.08 mmol), 2-phenoxyethyl-4-methylbenzenesulfonate (630 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieve (500 mg), DMF (6 ml), nitrogen protection, placed in 70 ℃ oil bath, stirring reaction 24 h. Transfer to 100 ml single-neck flask, rinse with ethanol, reduce pressure and concentrate, get orange oil, then redissolve with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:5), suction filtration, wash the filter cake with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:5), reduce pressure and concentrate, get 400 mg of brown oily mixture, without purification, directly to the next step.

[0439] 2, (3Z, 6Z)-3-(3-(4-fluorophenoxy)phenyl)methylene)-6-((5-isopropyl-l-(2- phenoxyethyl)-lH-imidazol-4-yl)methylene)piperazine-2, 5-dione

[0440]

[0441] Take 25 mL dry reaction bottle, (Z)-1-acetyl-3-((5-isopropyl-l-(2- phenoxyethyl)-lH-imidazol-4-yl)methylene)piperazine-2, 5-dione (300 mg, 0.76 mmol), 3-(4-fluorophenoxy)benzaldehyde (164 mg, 0.76 mmol), cesium carbonate (371 mg, 1.14 mmol), anhydrous sodium sulfate (215 mg, 1.52 mmol), DMF (6 mL), replace nitrogen protection, avoid light, put in 45 °C oil bath, stir for 20 h. Transfer to 100 ml single port bottle, ethanol rinse, concentrated under reduced pressure, then dissolved with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), suction filtration, the filter cake was washed with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), and concentrated under reduced pressure. Flash purification, (methanol: 87%, water: 13%) to obtain the product, concentrated under reduced pressure to obtain 168 mg of light yellow solid, the yield was 40.10%.

[0442] 1 H NMR (500 MHz, DMSO-d6) δ 11.97 (s, 1H), 10.13 (s, 1H), 7.96 (s, 1H), 7.41 (t, J = 7.8 Hz, 1H), 7.28 (dd, J = 14.2, 7.3 Hz, 3H), 7.23 (t, J = 8.6 Hz, 2H), 7.17 (s, 1H), 7.14-7.09 (m, 2H), 7.00-6.85 (m, 4H), 6.72 (s, 2H), 4.45 (s, 2H), 4.24 (s, 2H), 3.20-3.14 (m, 1H), 1.35 (d, J = 6.8 Hz, 2H). MS (ESI): m / z 553.02 [M + H] + Mp: 159-161 °C.

[0443] Example 58 Preparation of (3Z, 6Z)-3-(3-(4-fluorophenoxy)phenyl)methylene)-6-((5-isopropyl-l-(2-(p- tolyloxy)ethyl)-lH-imidazol-4-yl)methylene)piperazine-2, 5-dione (PLN-3-58)

[0444] 1, 2-(p-Tolyl oxy) ethyl-1-ol

[0445]

[0446] Take 50 ml dry round-bottom flask, p-methyl benzyl bromide (2 g, 11.69 mmol), potassium carbonate (4.85 g, 35.08 mmol), copper chloride (99 mg, 0.58 mmol), ethylene glycol (16 ml), nitrogen protection, placed in 130 ℃ oil bath, stirring reaction 12 h. Cool to room temperature, dilute hydrochloric acid pH value is 3, ethyl acetate (100 ml*2) extraction, combined organic phase, anhydrous sodium sulfate drying, filtration, reduced pressure concentration, column chromatography purification, PE:EA = 1:1, get white solid 730 mg, yield 41.10%.

[0447] 2, 2-(p-tolyloxy) ethyl-4-methyl benzene sulfonate

[0448]

[0449] Take 50 ml dry round-bottom flask, 2-(p-tolyloxy) ethyl-1-alcohol (1 g, 6.58 mmol) is dissolved in 10 ml anhydrous DCM, add triethylamine (3.32 g, 32.90 mmol), nitrogen protection, drop p-methyl benzyl sulfuryl chloride (2.25 g, 11.84 mmol) dissolved in 10 ml anhydrous DCM, transfer to room temperature, stirring reaction 6 h. Purification water (100 ml*2) wash, combined organic phase, anhydrous sodium sulfate drying, filtration, reduced pressure concentration, column chromatography purification, PE:EA = 4:1, get white solid 1.65 g, yield 81.96%.

[0450] 3, (Z)-1-acetyl-3-((5-isopropyl-1-(2-(p-tolyloxy) ethyl)-1H-imidazol-4-yl) methylene) piperazine-2, 5-dione

[0451]

[0452] Into a 50 mL dry round bottom flask, (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), 2-(p-tolyloxy)ethyl-4-methylbenzenesulfonate (660 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieves (500 mg), DMF (6 mL), nitrogen protection, placed in 70 °C oil bath, stirred for 24 h. Transfer to a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure, obtain orange oil, redissolve with a mixture of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:5), suction filtration, wash the filter cake with a mixture of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:5), concentrate under reduced pressure, obtain 400 mg of brown oil mixture, without purification, directly to the next step.

[0453] 2, (3Z, 6Z)-3-(3-(4-fluorophenoxy)phenyl)methylene)-6-((5-isopropyl-1-(2-(p-tolyloxy)ethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0454]

[0455] Into a 25 mL dry reaction flask, (Z)-1-acetyl-3-((5-isopropyl-1-(2-(p-tolyloxy)ethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.73 mmol), 3-(4-fluorophenoxy)benzaldehyde (158 mg, 0.73 mmol), cesium carbonate (357 mg, 1.09 mmol), anhydrous sodium sulfate (207 mg, 1.46 mmol), DMF (6 mL), replace nitrogen protection, avoid light, placed in 45 °C oil bath, stirred for 20 h. Transfer to a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure, redissolve with a mixture of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:3), suction filtration, wash the filter cake with a mixture of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:3), concentrate under reduced pressure. Flash purification, methanol: 90%, water: 10% to obtain the product, concentrate under reduced pressure, obtain 150 mg of light yellow solid, yield 36.32%.

[0456] 1H NMR (500 MHz, DMSO-d6) δ 11.97 (s, 1H), 10.13 (s, 1H), 7.95 (s, 1H), 7.40 (t, J = 7.9 Hz, 1H), 7.26 (d, J = 7.8 Hz, 1H), 7.23 (t, J = 8.7 Hz, 2H), 7.17 (s, 1H), 7.12 (dd, J = 8.0, 3.9 Hz, 2H), 7.07 (d, J = 7.8 Hz, 2H), 6.90 (d, J = 8.1 Hz, 1H), 6.81 (d, J = 7.9 Hz, 2H), 6.72 (s, 2H), 4.43 (s, 2H), 4.19 (s, 2H), 3.31 - 3.26 (m, 1H), 2.21 (s, 3H), 1.34 (d, J = 6.9 Hz, 6H). MS (ESI): m / z 567.63 [M+H] + Mp: 157-160 °C.

[0457] Example 59 Preparation of (3Z,6Z)-3-(3-(4-fluorophenoxy)phenyl)methylene-6-((5-isopropyl- 1-(2-(3-phenoxypropyl))-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-59)

[0458] 1, 3-Phenoxypropyl-1-alcohol

[0459]

[0460] Take 50 ml dry round bottom flask, in turn, bromobenzene (2 g, 12.74 mmol), potassium carbonate (5.28 g, 38.22 mmol), copper chloride (108 mg, 0.64 mmol), 1,3-propanediol (16 ml), nitrogen protection, placed in 130 °C oil bath, stirring reaction 12 h. Cool to room temperature, dilute hydrochloric acid to pH 3, ethyl acetate (100 ml*2) extraction, combined organic phase, anhydrous sodium sulfate drying, filtration, reduced pressure concentration, column chromatography purification, PE:EA = 4:1, get 1.1 g of light yellow oil liquid, yield 56.70%.

[0461] 2, 3-Phenoxypropyl 4-methylbenzenesulfonate

[0462]

[0463] Take 50 ml dry round-bottom flask, 0 ℃, 3-phenoxypropyl-1-alcohol (1 g, 6.58 mmol) was dissolved in 10 ml of anhydrous DCM, triethylamine (3.32 g, 32.90 mmol) was added, and nitrogen was protected. 2.25 g of p-methylbenzenesulfonyl chloride (11.84 mmol) dissolved in 10 ml of anhydrous DCM was added dropwise, and the reaction was stirred at room temperature for 6 h. Purification was performed with water (100 ml x 2) washing, and the combined organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography with PE:EA = 10:1 to obtain 1.60 g of white solid with a yield of 79.46%.

[0464] 3, (Z)-1-acetyl-3-((5-isopropyl-1-(3-phenoxypropyl)-1H-imidazol-4-yl) methylene) piperazine-2, 5-dione

[0465]

[0466] Take 50 ml dry round-bottom flask, and sequentially add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl) methylene) piperazine-2, 5-dione (300 mg, 1.08 mmol), 3-phenoxypropyl 4-methylbenzenesulfonate (660 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieve (500 mg), DMF (6 ml), and nitrogen protection. Place in a 70 ℃ oil bath and stir for 24 h. Transfer to a 100 ml single-neck flask, rinse with ethanol, and concentrate under reduced pressure to obtain orange oil. Redissolve with a mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:5), and extract with a mixed solvent (V ethanol:V dichloromethane = 1:5). Concentrate under reduced pressure to obtain a brown oily mixture 400 mg, which is not purified and directly subjected to the next step.

[0467] 2, (3Z, 6Z)-3-(3-(4-fluorophenoxy) phenyl) methylene-6-((5-isopropyl-1-(2-(3-phenoxypropyl))-1H-imidazol-4-yl) methylene) piperazine-2, 5-dione

[0468]

[0469] Take 25 mL dry reaction bottle, (Z)-1 -acetyl-3-((5-isopropyl-1 -(3- phenoxypropyl)-1 H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.73 mmol), 3-(4-fluorophenoxy)benzaldehyde (158 mg, 0.73 mmol), cesium carbonate (357 mg, 1.09 mmol), anhydrous sodium sulfate (207 mg, 1.46 mmol), DMF (6 mL), replace nitrogen protection, avoid light, put in 45 °C oil bath, stir for 20 h. Transfer to 100 ml single mouth bottle, ethanol rinse, concentrate under reduced pressure, then redissolve with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1 :3), suction filtration, wash the filter cake with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1 :3), concentrate under reduced pressure. Flash purification, methanol: 87%, water: 13% to get the product, concentrate under reduced pressure to get 168 mg of light yellow solid, yield 40.67%.

[0470] 1 H NMR (500 MHz, DMSO-d6) δ 11.81 (s, 1 H), 10.13 (s, 1 H), 7.84 (s, 1 H), 7.54 (s, 1 H), 7.43 (d, J = 7.5 Hz, 1 H), 7.33 (t, J = 7.9 Hz, 1 H), 7.29 (t, J = 7.7 Hz, 2H), 7.21 (t, J = 8.6 Hz, 2H), 7.08 (dd, J = 8.7, 4.5 Hz, 2H), 6.96 - 6.90 (m, 3H), 6.79 (d, J = 8.0 Hz, 1 H), 6.56 (s, 1 H), 6.53 (s, 1 H), 4.18 (t, J = 7.0 Hz, 2H), 3.98 (t, J = 5.7 Hz, 2H), 3.25 - 3.12 (m, 1 H), 2.24 - 2.04 (m, 2H), 1.29 (d, J = 7.0 Hz, 6H). MS (ESI): m / z 567.26 [M+H] + Mp: 178-181 °C.

[0471] Example 60 Preparation of (3Z,6Z)-3-(3-(4-fluorophenoxy)phenyl)methylene-6-((5- isopropyl-1 -(2-(2-(benzyloxy)ethyl)-1 H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-60)

[0472] 1,2-(benzyloxy)ethane-1 -ol

[0473]

[0474] Take 50ml dry round-bottom flask, add p-bromobenzyl (2g, 11.69mmol), potassium carbonate (4.85g, 35.08mmol), copper chloride (99mg, 0.58mmol), ethylene glycol (16ml) in turn, nitrogen protection, placed in 130℃ oil bath, stirring reaction 12h. Cool to room temperature, adjust pH to 3 with dilute hydrochloric acid, extract with ethyl acetate (100ml*2), combine the organic phase, dry over anhydrous sodium sulfate, filter, reduce pressure concentration, column chromatography purification, PE:EA = 10:1, get white solid 700mg, yield 39.41%.

[0475] 2, 2-(benzyloxy) ethyl-4-methylbenzenesulfonate

[0476]

[0477] Take 50ml dry round-bottom flask, 0℃, 2-(benzyloxy) ethyl-1-ol (1g, 6.58mmol) is dissolved in 10ml anhydrous DCM, add triethylamine (3.32g, 32.90mmol), nitrogen protection, drop p-methylbenzenesulfonyl chloride (2.25g, 11.84mmol) dissolved in 10ml anhydrous DCM, transfer to room temperature, stirring reaction 6h. Purification water (100ml*2) wash, combine the organic phase, dry over anhydrous sodium sulfate, filter, reduce pressure concentration, column chromatography purification, PE:EA = 4:1, get white solid 1.60g, yield 79.46%.

[0478] 3, (Z)-1-acetyl-3-((1-(2-(benzyloxy) ethyl)-5-isopropyl-1H-imidazol-4-yl) methylene) piperazine-2, 5-dione Take 50ml dry round-bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl) methylene) piperazine-2, 5-dione (300mg, 1.08mmol), 2-(benzyloxy) ethyl-4-methylbenzenesulfonate (660mg, 2.16mmol), cesium carbonate (704mg, 2.16mmol), potassium iodide (180mg, 1.08mmol), 4A molecular sieve (500mg), DMF (6ml), nitrogen protection, placed in 70℃ oil bath, stirring reaction 24h. Transfer to 100ml single neck flask, rinse with ethanol, reduce pressure concentration, get orange oil, then redissolve with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:5), suction filtration, wash the filter cake with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:5), reduce pressure concentration, get brown oil mixture 400mg, without purification, directly to the next step.

[0479] 2. (3Z, 6Z)-3-(3-(4-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(2-(2- (benzyloxy)ethyl)-1H-imidazol-4-yl)methyl)piperazin-2,5-dione

[0480]

[0481] Into a 25 mL dry reaction vial, (Z)-1-acetyl-3-((1-(2-(benzyloxy)ethyl)-5- isopropyl-1H-imidazol-4-yl)methyl)piperazin-2,5-dione (300 mg, 0.73 mmol), 3- (4-fluorophenoxy)benzaldehyde (158 mg, 0.73 mmol), cesium carbonate (357 mg, 1.09 mmol), anhydrous sodium sulfate (207 mg, 1.46 mmol), DMF (6 mL), replace nitrogen protection, avoid light, put into 45 °C oil bath, stir for 20 h. Transfer into 100 mL single port bottle, rinse with ethanol, concentrate under reduced pressure, redissolve with mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:3), suction filtration, wash the filter cake with mixed solvent (Vethanol:Vdichloromethane = 1:3), concentrate under reduced pressure. Flash purification, methanol: 80%, water: 20% to get the product, concentrate under reduced pressure to get 150 mg of light yellow solid, with a yield of 36.32%.

[0482] 1 H NMR (500 MHz, DMSO-d6) δ 12.00 (s, 1H), 10.13 (s, 1H), 7.88 (s, 1H), 7.41 (t, J = 7.9 Hz, 1H), 7.31 (t, J = 7.1 Hz, 2H), 7.29-7.25 (m, 2H), 7.22 (d, J = 6.0 Hz, 4H), 7.18 (s, 1H), 7.12 (dd, J = 8.3, 4.5 Hz, 2H), 6.90 (d, J = 8.0 Hz, 1H), 6.72 (s, 1H), 6.70 (s, 1H), 4.49 (s, 2H), 4.24 (s, 2H), 3.67 (s, 2H), 3.25-3.19 (m, 1H), 1.26 (d, J = 7.0 Hz, 6H). MS (ESI): m / z 567.90 [M+H] + Mp: 129-131 °C.

[0483] Example 61 Preparation of (3Z, 6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5- isopropyl-1-(3-piperidinyl)propylimidazol-4-yl)methyl)piperazin-2,5-dione (PLN-3-61)

[0484]

[0485] 1) Take 250 ml dry round bottom flask, add piperidine (3.0 g, 35.23 mmol), potassium carbonate (7.29 g, 52.75 mmol), potassium iodide (1.17 mg, 7.05 mmol), acetonitrile (120 ml), 1-chloro-3-bromopropane (6.66 g, 42.30 mmol) in sequence, stir the reaction in 25 °C oil bath for 16 h, monitor the reaction by TLC (MeOH:DCM = 1:10), the reaction is complete. Filter, filter cake EA wash, reduce pressure concentration, get light brown oil 2.61 g, yield: 45.79%.

[0486] 2) Take 50 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2,5-dione (300 mg, 1.09 mmol), cesium carbonate (707.55 mg, 2.17 mmol), potassium iodide (180.24 mg, 1.09 mmol), 4A molecular sieve (500 mg), N-3-chloropropylpiperidine (263.31 mg, 1.63 mmol) in DMF (5 ml) solution in sequence, exhaust, nitrogen protection, placed in 70 °C oil bath, stir the reaction for 24 h. After the reaction, naturally cool to room temperature, add 3-p-fluorobenzoylbenzaldehyde (247.80 mg, 1.09 mmol) in DMF (2 ml), warm up to 45 °C oil bath, stir the reaction for 24 h. Monitor the reaction by LC-MS, after the reaction, drop the reaction into 4 °C cold water (80 ml), suction filter, filter cake cold water wash, suction dry, dissolve in methanol and dichloromethane (1:3), filter, dry, methanol ultrasonic beating, placed in -30 °C refrigerator, suction filter, filter cake with cold methanol wash, 50 °C vacuum drying, get yellow solid 142.2 mg, yield 22.90%.

[0487] 1 H NMR (400 MHz, DMSO-d6)) δ 12.02 (s, 1H), 10.36 (s, 1H), 7.84 (dd, J = 19.67, 39.85 Hz, 5H), 7.61 (d, J = 9.85 Hz, 2H), 7.40 (s, 2H), 6.80 (s, 1H), 6.70 (s, 1H), 4.03 (s, 2H), 3.43 (s, 4H), 3.26 (s, 1H), 2.24 (d, J = 34.68 Hz, 8H), 1.82 (s, 2H), 1.49 (s, 6H). MS (ESI): m / z 570.27 [M+H] + Mp: 195-197 °C.

[0488] Example 62 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(3-piperidinyl)propylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-62)

[0489] 1, 1-(3-chloropropyl)piperidine

[0490]

[0491] Into a 250ml dry round bottom flask, piperidine (1.0g, 11.74mmol), potassium carbonate (2.91g, 17.61mmol), potassium iodide (195mg, 1.17mmol), acetonitrile (15ml), 1-chloro-3-bromopropane (1.85g, 11.74mmol) were added in sequence, and stirred in a 25°C oil bath for 5h. TLC (MeOH:DCM=1:10) was used to monitor the reaction until completion. The reaction mixture was extracted with ethyl acetate (50ml*3), and the organic phase was combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 1.20g of colorless oily liquid, with a yield of 63.49%.

[0492] 2, (Z)-1-acetyl-3-((5-isopropyl-1-(3-(piperidin-1-yl)propyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0493] Into a 50ml dry round bottom flask, (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300mg, 1.08mmol), 1-(3-chloropropyl)piperidine (348mg, 2.16mmol), cesium carbonate (704mg, 2.16mmol), potassium iodide (180mg, 1.08mmol), 4A molecular sieves (500mg), DMF (6ml) were added in sequence, and degassed, protected by nitrogen, and stirred in a 70°C oil bath for 24h. The reaction mixture was transferred into a 100ml single-neck flask, washed with ethanol, and concentrated under reduced pressure to obtain orange oily substance. The oily substance was redissolved in a mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane=1:5), filtered, and the filter cake was washed with a mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane=1:5). The filtrate was concentrated under reduced pressure to obtain 400mg of orange oily mixture, which was used directly in the next step without purification.

[0494]

[0495] 2, (3Z,6Z)-3-(3-(p-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(3-piperidinyl)propylimidazol-4-yl)methylene)piperazine-2,5-dione

[0496] ​

[0497] Take 25 mL dry reaction bottle, (Z)-1-acetyl-3-((5-isopropyl-1-(3-(piperidin-1- yl)propyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.75 mmol), 3- (4-fluorophenoxy)benzaldehyde (194 mg, 0.90 mmol), DMF (6 mL), cesium carbonate (367 mg, 1.12 mmol), anhydrous sodium sulfate (213 mg, 1.50 mmol), replace nitrogen protection, avoid light, put in 45 ℃ oil bath and stir for 20 h. Transfer to 100 ml single port bottle, ethanol rinse, concentrate under reduced pressure, and then redissolve with a mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), filter, wash the filter cake with a mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), and concentrate under reduced pressure. Add methanol (3 mL) and ultrasonic dispersion, and then stand at -20 ℃ overnight. Filter and dry to obtain 85 mg of yellow solid with a yield of 20.43%.

[0498] 1 H NMR (500 MHz, DMSO-d6) δ 12.00 (s, 1H), 10.12 (s, 1H), 7.87 (s, 1H), 7.41 (t, J = 7.9 Hz, 1H), 7.27 (d, J = 7.8 Hz, 1H), 7.23 (t, J = 8.8 Hz, 2H), 7.17 (s, 1H), 7.15-7.09 (m, 2H), 6.90 (dd, J = 8.1, 1.7 Hz, 1H), 6.71 (s, 2H), 4.03 (t, J = 7.0 Hz, 2H), 3.26-3.20 (m, 1H), 2.28 (s, 4H), 2.20 (t, J = 6.7 Hz, 2H), 1.88-1.73 (m, 2H), 1.47-1.51 (m, 4H), 1.37 (s, 2H), 1.33 (d, J = 7.1 Hz, 6H). MS (ESI): m / z 558.27 [M+H] + Mp: 166-168 ℃.

[0499] Example 63 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(3-pyrrolidinyl)propylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-63)

[0500] 1, 1-(3-chloropropyl)pyrrolidine

[0501]

[0502] Into a 250 ml dry round bottom flask, pyrrolidine (1.0 g, 14.06 mmol), potassium carbonate (2.91 g, 14.06 mmol), potassium iodide (233 mg, 1.41 mmol), acetonitrile (18 ml), 1-chloro-3-bromopropane (2.21 g, 14.06 mmol) were added in sequence, and stirred in a 25 °C oil bath for 12 h. The reaction was monitored by TLC (MeOH:DCM = 1:10) until completion. The reaction mixture was extracted with ethyl acetate (50 ml*3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a colorless oily liquid 1.25 g with a yield of 60.68%.

[0503] 2, (Z)-1-acetyl-3-((5-isopropyl-1-(3-(pyrrolidin-1-yl)propyl)-1H-imidazol-4-yl)methylene)piperazine-2, 5-dione

[0504]

[0505] Into a 50 ml dry round bottom flask, (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2, 5-dione (300 mg, 1.08 mmol), 1-(3-chloropropyl)pyrrolidine

[0506] (319 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieves (500 mg), DMF (6 ml), evacuated, nitrogen protection, placed in a 70 °C oil bath, stirred for 24 h. Transfer to a 100 ml single neck flask, rinse with ethanol, concentrate under reduced pressure to obtain orange oily substance, then redissolve with a mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:5), suction filtration, wash the filter cake with a mixed solvent (Vethanol:Vdichloromethane = 1:5), concentrate under reduced pressure to obtain an orange oily mixture 400 mg, which was not purified and directly subjected to the next step.

[0507] 3, (3Z, 6Z)-3-(3-(p-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(3-pyrrolidinyl)propyl imidazol-4-yl)methylene)piperazine-2, 5-dione

[0508]

[0509] Take 25 mL dry reaction bottle, (Z)-1 -acetyl-3-((5-isopropyl-1 -(3-(pyrrolidin-1 - yl)propyl)-1 H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.78 mmol), 3-(4- fluorophenoxy)benzaldehyde (168 mg, 0.78 mmol), DMF (6 mL), cesium carbonate (381 mg, 1.17 mmol), anhydrous sodium sulfate (221 mg, 1.56 mmol), replace nitrogen protection, avoid light, put in 45 °C oil bath, stir for 20 h. Transfer to 100 ml single port bottle, ethanol rinse, concentrate under reduced pressure, then redissolve with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), suction filtration, and the filter cake is washed with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), and concentrated under reduced pressure. Add methanol (3 mL), ultrasonic dispersion, -20 °C overnight. Filter, dry, get yellow solid 110 mg, yield 26.01%.

[0510] 1 H NMR (500 MHz, DMSO-d6) δ 12.00 (s, 1H), 10.12 (s, 1H), 7.87 (s, 1H), 7.41 (t, J = 7.9 Hz, 1H), 7.27 (d, J = 7.7 Hz, 1H), 7.23 (t, J = 8.8 Hz, 2H), 7.18 (s, 1H), 7.14-7.10 (m, 2H), 6.90 (dd, J = 8.1, 1.9 Hz, 1H), 6.70 (s, 2H), 4.05 (t, J = 7.1 Hz, 2H), 3.26-3.20 (m, 1H), 2.41 (s, 4H), 2.36 (t, J = 6.8 Hz, 2H), 1.89-1.79 (m, 2H), 1.68 (s, 4H), 1.33 (d, J = 7.1 Hz, 6H). MS (ESI): m / z 544.24 [M+H] + Mp: 172-174 °C.

[0511] Example 64 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1- (dihydroindol-1-yl)propyl imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-64)

[0512] 1, 1 -(3-chloropropyl)dihydroindole

[0513]

[0514] Into a 100 ml dry round bottom flask, indoline (1.0 g, 8.39 mmol), cesium carbonate (4.1 g, 12.58 mmol), potassium iodide (279 mg, 1.68 mmol), acetonitrile (15 ml), 1-chloro-3-bromopropane (2.64 g, 16.78 mmol) were added successively and stirred at 25 °C oil bath for 20 h. The reaction was monitored by TLC (EA:PE = 1:20) until completion. The reaction mixture was extracted with ethyl acetate (50 ml*2), the organic phase was combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (PE:EA = 100:1) to give a yellowish oil 0.90 g with a yield of 54.88%.

[0515] 3. (Z)-1-acetyl-3-((1-(3-(dihydroindol-1-yl)propyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0516]

[0517] Into a 50 ml dry round bottom flask, (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), 1-(3-chloropropyl)dihydroindole (421 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieves (500 mg), DMF (6 ml) were added successively, evacuated, and protected by nitrogen. The reaction was stirred at 70 °C oil bath for 24 h. The reaction mixture was transferred into a 100 ml single-neck flask, washed with ethanol, and concentrated under reduced pressure to give an orange oil. The orange oil was redissolved in a mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:5), filtered, and the filter cake was washed with a mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:5). The filtrate was concentrated under reduced pressure to give a brown oil 450 mg, which was used directly in the next step without purification.

[0518] 4. (3Z,6Z)-3-(3-(p-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(dihydroindol-1-yl)propyl-imidazol-4-yl)methylene)piperazine-2,5-dione

[0519]

[0520] Take 25 mL dry reaction bottle, (Z)-1 -acetyl-3-((1 -(3-(dihydroindol-1 - yl)propyl)-5-isopropyl-1 H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.69 mmol), 3-(4-fluorophenoxy)benzaldehyde (149 mg, 0.69 mmol), cesium carbonate (339 mg, 1.04 mmol), anhydrous sodium sulfate (196 mg, 1.38 mmol), DMF (6 mL), replace nitrogen protection, avoid light, put in 45 °C oil bath, stir for 20 h. Transfer to 100 ml single port bottle, ethanol rinse, water, concentrated under reduced pressure, and then dissolved in a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1 :3), suction filtration, and then washed the filter cake with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1 :3), and then concentrated under reduced pressure. Flash purification, methanol: 81 %, water: 29 % to obtain the product, concentrated under reduced pressure to obtain 128 mg of light yellow solid, yield 31.45 %.

[0521] 1 H NMR (500 MHz, DMSO-d6) δ 12.00 (s, 1 H), 10.13 (s, 1 H), 7.92 (s, 1 H), 7.41 (t, J = 7.8 Hz, 1 H), 7.27 (d, J = 7.8 Hz, 1 H), 7.23 (t, J = 8.4 Hz, 2H), 7.17 (s, 1 H), 7.13 (d, J = 4.0 Hz, 2H), 7.03 (d, J = 6.8 Hz, 1 H), 7.00-6.96 (m, 1 H), 6.90 (d, J = 7.5 Hz, 1 H), 6.71 (d, J = 5.8 Hz, 2H), 6.57 (t, J = 7.2 Hz, 1 H), 6.45 (d, J = 7.6 Hz, 1 H), 4.12 (s, 2H), 3.28 (t, J = 7.5 Hz, 2H), 3.25-3.20 (m, 1 H), 3.06 (t, J = 6.2 Hz, 2H), 2.88 (t, J = 7.8 Hz, 2H), 2.00-1.96 (m, 2H), 1.32 (d, J = 6.7 Hz, 6H). MS (ESI): m / z 592.43 [M+H] + Mp: 143-145 °C.

[0522] Example 65 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1 -(dihydroisoquinolin-2-yl)propyl imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-65)

[0523] 1,2-(3-chloropropyl)-1,2,3,4-tetrahydroisoquinoline

[0524]

[0525] Into a 100 ml dry round bottom flask, 1,2,3,4-tetrahydroisoquinoline (1.0 g, 7.50 mmol), cesium carbonate (3.66 g, 11.25 mmol), potassium iodide (124 mg, 0.75 mmol), acetonitrile (10 ml), 1-chloro-3-bromopropane (2.36 g, 15.00 mmol) were added successively and stirred at 25 °C oil bath for 20 h. The reaction was monitored by TLC (EA:PE = 1:10) until completion. The reaction mixture was extracted with ethyl acetate (50 ml*2), the organic phase was combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (PE:EA = 100:1) to give a yellowish oil 220 g with a yield of 14.01%.

[0526] 3, (Z)-1-acetyl-3-((1-(3-(3,4-dihydroisoquinolin-2(1H)-yl)propyl)-5-isopropyl-1H- imidazol-4-yl)methylene)piperazine-2,5-dione

[0527]

[0528] Into a 50 ml dry round bottom flask, (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), 2-(3-chloropropyl)-1,2,3,4- tetrahydroisoquinoline (451 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieves (500 mg), DMF (6 ml) were added successively, evacuated, and protected by nitrogen. The reaction mixture was stirred at 70 °C oil bath for 24 h. The reaction mixture was transferred into a 100 ml single-neck flask, washed with ethanol, and concentrated under reduced pressure to give an orange oil. The orange oil was redissolved in a mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:5), filtered, and the filter cake was washed with a mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:5). The filtrate was concentrated under reduced pressure to give a brown oil 450 mg, which was used directly in the next step without purification.

[0529] 4, (3Z,6Z)-3-(3-(p-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(dihydroisoquinolin-2- yl)propylimidazol-4-yl)methylene)piperazine-2,5-dione

[0530]

[0531] Into a 25 mL dry reaction vial, (Z)-1-acetyl-3-((1-(3-(3,4-dihydroisoquinolin-2(1H)- yl)propyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0532] Into a 25 mL dry reaction vial, (Z)-1-acetyl-3-((1-(3-(3,4-dihydroisoquinolin-2(1H)- yl)propyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0533] 1 H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.13 (s, 1H), 7.93 (s, 1H), 7.41 (t, J = 7.9 Hz, 1H), 7.27 (d, J = 8.0 Hz, 1H), 7.23 (t, J = 8.8 Hz, 2H), 7.17 (s, 1H), 7.14 - 7.08 (m, 2H), 6.95 - 6.88 (m, 2H), 6.86 (d, J = 7.3 Hz, 1H), 6.72 (s, 1H), 6.69 (s, 1H), 6.49 (d, J = 8.2 Hz, 1H), 6.46 (t, J = 7.3 Hz, 1H), 4.09 (t, J = 7.3 Hz, 2H), 3.28 (t, J = 7.5 Hz, 2H), 3.23 - 3.19 (m, 3H), 2.66 (t, J = 6.2 Hz, 2H), 1.97 - 1.89 (m, 2H), 1.87 - 1.82 (m, 2H), 1.30 (d, J = 7.1 Hz, 6H). MS (ESI): m / z 606.48 [M+H] + Mp: 166-169 °C.

[0534] Example 66 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(1H- pyrrol-1-yl)propyl)imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-66)

[0535] 1,1-(3-chloropropyl)-1H-pyrrole

[0536]

[0537] Into a 100 ml dry round bottom flask, 1H-pyrrole (1.0 g, 14.90 mmol), cesium carbonate (7.28 g, 22.35 mmol), potassium iodide (247 mg, 1.49 mmol), acetonitrile (15 ml), 1-chloro-3-bromopropane (2.35 g, 14.90 mmol) were added in sequence, and the reaction was stirred in a 25 °C oil bath for 20 h. TLC (EA: PE = 1:20) was used to monitor the reaction until completion. The reaction was extracted with ethyl acetate (50 ml*2), and the organic phase was combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 0.64 g of a light yellow oily liquid with a yield of 30.05%.

[0538] 3, (Z)-3-((1-(3-(1H-pyrrol-1-yl)propyl)-5-isopropyl-1H-imidazol-4-yl)methylene)-1- acetyl piperazine-2, 5-dione

[0539]

[0540] Into a 50 ml dry round bottom flask, (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2, 5-dione (300 mg, 1.08 mmol), 1-(3-chloropropyl)-1H-pyrrole (309 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieves (500 mg), DMF (6 ml) were added in sequence, and the reaction was stirred in a 70 °C oil bath for 24 h. The reaction was transferred to a 100 ml single-neck flask, washed with ethanol, and concentrated under reduced pressure with a water, gas, and oil pump to obtain an orange oily substance. The oily substance was redissolved in a mixed solvent of ethanol and dichloromethane (Vethanol: Vdichloromethane = 1:5), filtered, and the filter cake was washed with a mixed solvent of ethanol and dichloromethane (Vethanol: Vdichloromethane = 1:5). The filtrate was concentrated under reduced pressure to obtain a brown oily mixture 400 mg, which was used directly in the next step without purification.

[0541] 4, (3Z, 6Z)-3-(3-(p-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(1H-pyrrol-1- yl)propyl imidazol-4-yl)methylene)piperazine-2, 5-dione

[0542]

[0543] Take 25 mL dry reaction bottle, (Z)-3-((1-(3-(1H-pyrrol-1-yl)propyl)-5- isopropyl-1H-imidazol-4-yl)methylene)-1-acetyl piperazine-2,5-dione (300 mg, 0.78 mmol), 3-(4-fluorophenoxy)benzaldehyde (203 mg, 0.94 mmol), cesium carbonate (381 mg, 1.17 mmol), anhydrous sodium sulfate (221 mg, 1.56 mmol), DMF (6 mL), replace nitrogen protection, avoid light, put in 45 °C oil bath and stir for 20 h. Transfer to 100 ml single port bottle, ethanol rinse, concentrate under reduced pressure, then redissolve with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), filter, wash the filter cake with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), and concentrate under reduced pressure. Flash purification, methanol: 70%, water: 30% to obtain the product, concentrate under reduced pressure to obtain 90 mg of light yellow solid, with a yield of 21.33%.

[0544] 1 H NMR (500 MHz, DMSO-d6) δ 11.97 (s, 1H), 10.13 (s, 1H), 7.87 (s, 1H), 7.41 (t, J = 7.9 Hz, 1H), 7.27 (d, J = 7.8 Hz, 1H), 7.23 (t, J = 8.6 Hz, 2H), 7.17 (s, 1H), 7.12 (dd, J = 8.0, 4.7 Hz, 2H), 6.90 (d, J = 8.1 Hz, 1H), 6.79 (s, 2H), 6.72 (s, 1H), 6.68 (s, 1H), 6.02 (d, J = 1.1 Hz, 2H), 3.95 (t, J = 8.5 Hz, 4H), 3.13 - 3.07 (m, 1H), 2.25 - 2.05 (m, 2H), 1.27 (d, J = 7.0 Hz, 6H). MS (ESI): m / z 540.17 [M+H] + .Mp: 190-192 °C.

[0545] Example 67 Preparation of (3Z,6Z)-3-(3-(4-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1-(1H-pyrrol-1-yl)propylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-67)

[0546] 1, (Z)-3-((1-(3-(1H-pyrrol-1-yl)propyl)-5-isopropyl-1H-imidazol-4-yl)methylene)-6-((Z)-3-(4- fluorobenzoyl)benzylidene) piperazine-2,5-dione

[0547]

[0548] Into a 25 mL dry reaction vial, (Z)-3-((1-(3-(1H-pyrrol-1-yl)propyl)-5- isopropyl-1H-imidazol-4-yl)methylene)-1-acetyl piperazine-2,5-dione (300 mg, 0.78 mmol), 3-(4-fluorobenzoyl)benzaldehyde (214 mg, 0.94 mmol), cesium carbonate (381 mg, 1.17 mmol), anhydrous sodium sulfate (221 mg, 1.56 mmol), DMF (6 mL), nitrogen protection, light protection, 45 °C oil bath, stirring reaction for 20 h. Transfer to a 100 mL single-mouth bottle, rinse with ethanol, concentrate under reduced pressure, and then redissolve with a mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:3), filter, and then rinse the filter cake with a mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:3), and concentrate under reduced pressure. Flash purification, methanol:80%, water:20%, to obtain the product, and concentrate under reduced pressure to obtain 66 mg of a light yellow solid, with a yield of 15.38%.

[0549] 1 H NMR (500 MHz, DMSO-d6) δ 12.00 (s, 1H), 10.35 (s, 1H), 7.90 (t, J = 7.5 Hz, 2H), 7.87 (s, 1H), 7.82 (s, 1H), 7.75 (d, J = 7.5 Hz, 1H), 7.63 (d, J = 7.6 Hz, 1H), 7.58 (t, J = 7.6 Hz, 1H), 7.40 (t, J = 8.6 Hz, 2H), 6.81 (s, 1H), 6.79 (s, 2H), 6.68 (s, 1H), 6.02 (s, 2H), 4.04-3.85 (m, 4H), 3.13-3.07 (m, 1H), 2.16-2.11 (m, 2H), 1.27 (d, J = 7.0 Hz, 6H) MS (ESI): m / z 552.22 [M+H] + Mp: 199-201 °C.

[0550] Example 68 Preparation of (3Z,6Z)-3-(3-(4-fluorobenzoyl)phenyl)methylene-6-((5- cyclopropyl-1-(1H-pyrrol-1-yl)propyl imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-68)

[0551] 1, (Z)-3-((1-(3-(1H-pyrrol-1-yl)propyl)-5-cyclopropyl-1H-imidazol-4-yl)methylene)-1- acetyl piperazine-2,5-dione

[0552]

[0553] Take 50ml dry round bottom flask, add ((Z)-1-acetyl-3-((5-cyclopropyl-1H- imidazol-4-yl)methylene)piperazine-2,5-dione (300mg, 1.09mmol), 1-(3- chloropropyl)-1H-pyrrole (311mg, 2.18mmol), cesium carbonate (710mg, 2.18mmol), potassium iodide (180mg, 1.09mmol), 4A molecular sieve (500mg), DMF (6ml), exhaust, nitrogen protection, placed in 70°C oil bath, stirring reaction for 24h. Transfer to 100ml single necked flask, ethanol rinse, reduced pressure concentration, get orange oil, then dissolved in a mixture of ethanol and dichloromethane (V ethanol: V dichloromethane = 1:5), suction filtration, mixed solvent (V ethanol: V dichloromethane = 1:5) wash the filter cake, reduced pressure concentration, get brown oil mixture 400mg, without purification, directly to the next step.

[0554] 2, (Z)-3-((1-(3-(1H-pyrrol-1-yl)propyl)-5-cyclopropyl-1H-imidazol-4-yl)methylene)-6- ((Z)-3-(4-fluorobenzoyl)benzylidene)piperazine-2,5-dione

[0555]

[0556] Take 25mL dry reaction bottle, (Z)-3-((1-(3-(1H-pyrrol-1-yl)propyl)-5-cyclopropyl-1H- imidazol-4-yl)methylene)-1-acetylpiperazine-2,5-dione (300mg, 0.78mmol), 3-(4- fluorobenzoyl)benzaldehyde (214mg, 0.94mmol), cesium carbonate (381mg, 1.17mmol), anhydrous sodium sulfate (221mg, 1.56mmol), DMF (6mL), replace nitrogen protection, avoid light, placed in 45°C oil bath under stirring reaction for 20h. Transfer to 100ml single necked flask, ethanol rinse, reduced pressure concentration, then dissolved in a mixture of ethanol and dichloromethane (V ethanol: V dichloromethane = 1:3), suction filtration, mixed solvent (V ethanol: V dichloromethane = 1:3) wash the filter cake, reduced pressure concentration. Flash purification, methanol: 80%, water: 20% when the product is obtained, reduced pressure concentration, get 100mg light yellow solid, yield 23.26%.

[0557] 1H NMR (500 MHz, DMSO-d6) δ 11.90 (s, 1H), 10.34 (s, 1H), 7.90 (d, J = 8.0 Hz, 3H), 7.82 (s, 1H), 7.75 (d, J = 7.3 Hz, 1H), 7.63 (d, J = 7.2 Hz, 1H), 7.58 (t, J = 7.5 Hz, 1H), 7.40 (t, J = 8.5 Hz, 2H), 6.80 (d, J = 1.9 Hz, 3H), 6.70 (s, 1H), 6.01 (s, 2H), 4.00 (t, J = 6.5 Hz, 1H), 3.96 (t, J = 6.5 Hz, 1H), 2.29 - 2.17 (m, 2H), 1.71 - 1.66 (m, 1H), 0.99 - 0.93 (m, 2H), 0.59 - 0.54 (m, 2H). MS (ESI): m / z 550.10 [M+H] + Mp: 166.2-168.3 °C.

[0558] Example 69 Preparation of (3Z,6Z)-3-(3-(4-fluorophenoxy)phenyl)methylene-6-((5- cyclopropyl-1-(1H-pyrrol-1-yl)propyl)imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-69)

[0559]

[0560] Take 25 mL dry reaction bottle, (Z)-3-((1-(3-(1H-pyrrol-1-yl)propyl)-5-cyclopropyl-1H- imidazol-4-yl)methylene)-1-acetylpiperazine-2,5-dione (300 mg, 0.78 mmol), 3-(4- fluorophenoxy)benzaldehyde (203 mg, 0.94 mmol), cesium carbonate (381 mg, 1.17 mmol), anhydrous sodium sulfate (221 mg, 1.56 mmol), DMF (6 mL), replace nitrogen protection, avoid light, put in 45 °C oil bath, stir for 20 h. Transfer to 100 ml single port bottle, ethanol rinse, reduce pressure concentration, then use mixed solvent (V ethanol:V dichloromethane = 1:3) to dissolve, suction filtration, mixed solvent (V ethanol:V dichloromethane = 1:3) to wash the filter cake, reduce pressure concentration. Flash purification, methanol:80%, water:20% to get the product, reduce pressure concentration, get 60 mg light yellow solid, yield is 14.33%.

[0561] 1H NMR (500 MHz, DMSO-d6) δ 11.88 (s, 1H), 10.13 (s, 1H), 7.91 (s, 1H), 7.41 (t, J = 7.9 Hz, 1H), 7.26 (d, J = 7.8 Hz, 1H), 7.23 (t, J = 8.8 Hz, 2H), 7.17 (s, 1H), 7.12 (dd, J = 8.4, 4.4 Hz, 2H), 6.90 (d, J = 8.0 Hz, 1H), 6.80 (s, 2H), 6.71 (d, J = 7.5 Hz, 2H), 6.01 (s, 2H), 4.00 (t, J = 7.3 Hz, 2H), 3.96 (t, J = 6.6 Hz, 2H), 2.26 - 2.20 (m, 2H), 1.76 - 1.56 (m, 1H), 0.99 - 0.94 (m, 2H), 0.58 - 0.52 (m, 2H). MS (ESI): m / z 538.32 [M+H] + Mp: 190.2-192.3 °C.

[0562] Example 70 Preparation of (3Z,6Z)-3-(3-(p-Fluorophenoxy)phenyl)methylene-6-((5- isopropyl-1-(1H-pyrrol-1-yl)butyl)imido) methylene)piperazine-2,5-dione (PLN-3-70)

[0563] 1) 1-(4-Chlorobutyl)-1H-pyrrole

[0564]

[0565] Into a 100 ml dry round bottom flask, 1H-pyrrole (1.0 g, 14.90 mmol), cesium carbonate (7.28 g, 22.35 mmol), potassium iodide (247 mg, 1.49 mmol), acetonitrile (15 ml), 1-chloro-4-bromobutane (2.55 g, 14.90 mmol) were added in sequence and stirred at 25 °C oil bath for 20 h, TLC (EA:PE = 1:10) was used to monitor the reaction until completion. The reaction mixture was extracted with ethyl acetate (50 ml*2), the organic phase was combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a light yellow oily liquid 0.50 g with a yield of 21.36%.

[0566] 2) (Z)-3-((1-(4-(1H-pyrrol-1-yl)butyl)-5-isopropyl-1H-imidazol-4-yl)methylene)-1- acetylpiperazine-2,5-dione

[0567]

[0568] Take 50 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), 1-(4-chlorobutyl)-1H-pyrrole (339 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieve (500 mg), DMF (6 ml), exhaust, nitrogen protection, placed in 70 °C oil bath, stirred for 24 h. Transfer to 100 ml single necked flask, ethanol rinse, concentrate under reduced pressure, get orange oil, then redissolve with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:5), suction filtration, the filter cake is washed with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:5), concentrate under reduced pressure, get brown oil mixture 400 mg, without purification, directly to the next step.

[0569] 3) (3Z,6Z)-3-(3-(p-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(1H-pyrrol-1- yl)butyl-imidazol-4-yl)methylene)piperazine-2,5-dione

[0570]

[0571] Take 25 mL dry reaction bottle, (Z)-3-((1-(4-(1H-pyrrol-1-yl)butyl)-5-isopropyl-1H- imidazol-4-yl)methylene)-1-acetylpiperazine-2,5-dione (300 mg, 0.76 mmol), 3-(4- fluorobenzoyl)benzaldehyde (196 mg, 0.91 mmol), cesium carbonate (371 mg, 1.14 mmol), anhydrous sodium sulfate (215 mg, 1.52 mmol), DMF (6 mL), nitrogen protection, light protection, placed in 45 °C oil bath, stirred for 20 h. Transfer to 100 ml single necked flask, ethanol rinse, concentrate under reduced pressure, then redissolve with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), suction filtration, the filter cake is washed with a mixture of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), concentrate under reduced pressure. Flash purification, methanol: 80%, water: 20% to get the product, concentrate under reduced pressure, get 80 mg light yellow solid, yield 13.31%.

[0572] 1H NMR (500 MHz, dmso) δ 11.98 (s, 1H), 10.09 (s, 1H), 7.86 (s, 1H), 7.41 (t, J = 7.9 Hz, 1H), 7.27 (d, J = 7.8 Hz, 1H), 7.23 (t, J = 8.7 Hz, 2H), 7.17 (s, 1H), 7.12 (dd, J = 8.6, 4.4 Hz, 2H), 6.90 (d, J = 7.9 Hz, 1H), 6.73 (s, 2H), 6.72 (s, 1H), 6.68 (s, 1H), 5.97 (s, 2H), 3.98 (t, J = 7.0 Hz, 2H), 3.91 (t, J = 6.6 Hz, 2H), 3.26 - 3.03 (m, 1H), 1.74 - 1.64 (m, 2H), 1.62 - 1.51 (m, 2H), 1.28 (d, J = 7.0 Hz, 6H). MS (ESI): m / z 554.45 [M+H] + Mp: 178-180 °C.

[0573] Example 71 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(1H-indol-1-yl)propyl)imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-71)

[0574] 1, 1-(3-chloropropyl)-1H-indole

[0575]

[0576] Take 100 ml dry two mouth round bottom flask, 0 °C, add 1H-indole (1.0 g, 8.54 mmol), sodium hydride (307 mg, 12.80 mmol), potassium iodide (142 mg, 0.85 mmol) in turn, drop DMF (18 ml), transfer to room temperature stirring 2 h, drop 1-chloro-3-bromopropane (2.02 g, 12.80 mmol), transfer to 25 °C oil bath under stirring reaction 12 h, TLC (EA: PE = 1:10) monitoring reaction to completion. Dichloromethane (50 ml*2) extraction, combined organic phase, anhydrous sodium sulfate drying, filtration, reduced pressure concentration, column chromatography purification, PE: EA = 80:1, colorless oil liquid 660 mg, yield 40%.

[0577] 3, (Z)-3-((1-(3-(1H-indol-1-yl)propyl)-5-isopropyl-1H-imidazol-4-yl)methylene)-1- acetylpiperazine-2,5-dione

[0578]

[0579] Into a 50 mL dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H- imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), 1-(3- chloropropyl)-1H-indole (416 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieves (500 mg), DMF (6 mL), evacuate, nitrogen protection, put into 70 °C oil bath, stir for 24 h. Transfer into a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure, get orange oil, resolubilize with mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:5), suction filtration, wash the filter cake with mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:5), concentrate under reduced pressure, get 450 mg of brown oil mixture, without purification, directly proceed to the next step.

[0580] 4, (3Z, 6Z)-3-(3-(p-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(1H- indol-1-yl)propyl)imidazol-4-yl)methylene)piperazine-2,5-dione

[0581]

[0582] Into a 25 mL dry reaction flask, add (Z)-3-((1-(3-(1H-indol-1-yl)propyl)-5- isopropyl-1H-imidazol-4-yl)methylene)-1-acetylpiperazine-2,5-dione (300 mg, 0.69 mmol), 3-(4-fluorophenoxy)benzaldehyde (149 mg, 0.69 mmol), cesium carbonate (339 mg, 1.04 mmol), anhydrous sodium sulfate (196 mg, 1.38 mmol), DMF (6 mL), replace nitrogen protection, avoid light, put into 45 °C oil bath, stir for 20 h. Transfer into a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure with water, gas, and oil pump, resolubilize with mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), suction filtration, wash the filter cake with mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), concentrate under reduced pressure. Flash purification, get the product when methanol: 81%, water: 29%, concentrate under reduced pressure, get 150 mg of light yellow solid, with a yield of 36.95%.

[0583] 1H NMR (500 MHz, DMSO-d6) δ 11.97 (s, 1H), 10.13 (s, 1H), 7.91 (s, 1H), 7.56 (d, J = 7.8 Hz, 1H), 7.46 (d, J = 8.2 Hz, 1H), 7.41 (d, J = 3.3 Hz, 1H), 7.39 (s, 1H), 7.26 (d, J = 7.9 Hz, 1H), 7.23 (t, J = 8.7 Hz, 2H), 7.17 (s, 1H), 7.15 - 7.09 (m, 3H), 7.03 (t, J = 7.3 Hz, 1H), 6.90 (d, J = 7.6 Hz, 1H), 6.71 (s, 1H), 6.66 (s, 1H), 6.46 (d, J = 2.4 Hz, 1H), 4.27 (t, J = 6.7 Hz, 2H), 4.01 (t, J = 7.5 Hz, 2H), 3.06 - 3.00 (m, 1H), 2.30 - 2.11 (m, 2H), 1.19 (d, J = 7.0 Hz, 6H). MS (ESI): m / z 590.67 [M+H] + Mp: 171-173 °C.

[0584] Example 72 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)phenyl)methylene-6-((5-isopropyl-1- (3-phenyl)propylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-72)

[0585]

[0586] A 25 ml dry round bottom flask was charged with (Z)-1-acetyl-3-((5-isopropyl-1H- imidazol-4-yl)methylene)piperazine-2,5-dione (200 mg, 0.72 mmol), cesium carbonate (471.65 mg, 1.45 mmol), potassium iodide (120.17 mg, 0.72 mmol), 4A molecular sieves (500 mg), 1-bromo-3-phenylpropane (216.17 mg, 1.09 mmol) in DMF (4 ml) solution, evacuated, nitrogen protection, placed in 70 °C oil bath, stirred for 24 h. After the reaction, it was naturally cooled to room temperature, and then 3-p-fluorobenzoylbenzaldehyde (132.17 mg, 0.58 mmol) in DMF (2 ml) was added, and the temperature was raised to 45 °C oil bath, and stirred for 18.5 h. The reaction was monitored by LC-MS, and after the reaction, the reaction solution was dropped into 4 °C cold water (70 ml), suction filtered, the filter cake was washed with cold water, suction dried, dissolved in methanol and dichloromethane (1:3), filtered, dried, methanol was ultrasonically beaten, placed in a -30 °C refrigerator, suction filtered, the filter cake was washed with cold methanol, and dried at 50 °C under vacuum to give a yellow solid 98.5 mg, with a yield of 24.18%.

[0587] 1 H NMR (600 MHz, DMSO) δ 12.03 (s, 1H), 10.35 (s, 1H), 7.94-7.89 (m, 3H), 7.82 (s, 1H), 7.75 (d, J = 7.72 Hz, 1H), 7.64 (d, J = 7.71 Hz, 1H), 7.59 (t, J = 7.65 Hz, 1H), 7.40 (dd, J = 5.31, 12.28 Hz, 2H), 7.30 (t, J = 7.56 Hz, 2H), 7.25-7.18 (m, 3H), 6.81 (s, 1H), 6.70 (s, 1H), 4.04 (t, J = 7.39 Hz, 2H), 3.22-3.13 (m, 1H), 2.61 (d, J = 7.98 Hz, 2H), 2.07-1.93 (m, 2H), 1.29 (d, J = 7.13 Hz, 6H). MS (ESI): m / z 563.09 [M+H] + Mp: 200-202 °C.

[0588] Example 73 Preparation of (3Z,6Z)-3-(3-(4-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(3- phenyl)propyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-3-73)

[0589] 1. (Z)-1-acetyl-3-((5-isopropyl-1-(3-phenylpropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0590]

[0591] Take 50 ml dry round bottom flask, add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4- yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), (3-chloropropyl)benzene (334 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieve (500 mg), DMF (6 ml) in turn, exhaust, nitrogen protection, put into 70 °C oil bath, stir for 24 h. Transfer to 100 ml single neck flask, rinse with ethanol, concentrate under reduced pressure, get orange oil, then redissolve with mixed solvent of ethanol and dichloromethane (Vethanol:Vdichloromethane = 1:5), suction filtration, mixed solvent (Vethanol:Vdichloromethane = 1:5) wash the filter cake, concentrate under reduced pressure, get brown oil mixture 400 mg, without purification, directly to the next step.

[0592] 2. (3Z, 6Z)-3-(3-(4-fluorophenoxy)phenyl)methylene-6-((5-isopropyl-1-(3- phenyl)propyl-1H-imidazol-4-yl)methylene)piperazine-2, 5-dione

[0593]

[0594] A 25 mL dry reaction flask was charged with (Z)-1-acetyl-3-((5-isopropyl-1-(3- phenylpropyl)-1H-imidazol-4-yl)methylene)piperazine-2, 5-dione (300 mg, 0.76 mmol), 3- (4-fluorophenoxy)benzaldehyde (164 mg, 0.76 mmol), cesium carbonate (371 mg, 1.14 mmol), anhydrous sodium sulfate (215 mg, 1.52 mmol), DMF (6 mL), and the reaction was stirred under a nitrogen atmosphere at 45 °C for 20 h. The reaction was transferred to a 100 mL single neck flask, rinsed with ethanol, concentrated under reduced pressure, and re-dissolved in a mixture of ethanol and dichloromethane (1:3). The solution was filtered, and the filter cake was rinsed with a mixture of ethanol and dichloromethane (1:3). The filtrate was concentrated under reduced pressure, and the product was purified by flash chromatography (methanol: 83%, water: 17%) to give a yellow solid. The product was concentrated under reduced pressure to give 150 mg of a yellow solid in 35.88% yield.

[0595] 1 H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.19 (s, 1H), 7.91 (s, 1H), 7.41 (t, J = 7.9 Hz, 1H), 7.29 (dd, J = 15.8, 8.2 Hz, 3H), 7.22 (d, J = 8.4 Hz, 4H), 7.19 (d, J = 5.5 Hz, 2H), 7.14 - 7.08 (m, 2H), 6.90 (dd, J = 8.1, 2.1 Hz, 1H), 6.71 (s, 1H), 6.69 (s, 1H), 4.04 (t, J = 7.3 Hz, 2H), 3.23 - 3.06 (m, 1H), 2.61 (t, J = 7.5, 1H), 2.06 - 1.95 (m, 2H), 1.29 (d, J = 7.1 Hz, 3H). MS (ESI): m / z 551.4 [M+H] + Mp: 161-164 °C.

[0596] Cell proliferation inhibition test of the compound of Effect Example 1

[0597] Tumor cells in log phase growth (NCI-H460, BxPC-3, HT-29) were trypsinized, centrifuged, and counted using a cell counting plate after resuspension in fresh media. 100 μL of media containing 2000-6000 cells per well were added to each well of a 96-well plate and the plate was incubated in an incubator (37°C, 5% CO2). After the cells were fully adherent, fresh media was used to dilute the test samples and Plinabulin to various concentrations, with 4 wells per concentration. After 72 hours of drug treatment, 20 μL of MTT at a concentration of 5 mg / L was added to each well and the plate was incubated for 3-4 hours. The media and MTT were then discarded and 100 μL of DMSO was added to each well. The plate was placed on a 96-well plate shaker to ensure complete dissolution of the purple formazan. The absorbance (OD value) was then measured using a microplate reader at a wavelength of 490 nm. Zeroing wells (blank media, MTT, DMSO) were also included in the experiment. Cell inhibition = 1 - (OD value of drug group - OD value of zeroing group) / (OD value of blank group - OD value of zeroing group) x 100%.

[0598] The results of the cell proliferation inhibition assay for the compounds of the present application are shown in Table 1, where "A" represents the calculated IC 50 less than 1 nM; "B" represents the calculated IC 50 from 1 nM to less than 10 nM; "C" represents the calculated IC 50 from 10 nM to less than 100 nM; "D" represents the calculated IC 50 greater than or equal to 100 nM.

[0599] Table 1 Cell proliferation inhibition assay for compounds of the present application

[0600]

[0601]

[0602]

[0603] Note: NCI-H460 is a human non-small cell lung cancer tumor cell line, BXPC-3 is a human pancreatic cancer tumor cell line, and HT-29 is a human colon cancer cell line. Plinabulin is the positive control. The control is DMSO without sample.

[0604] Compound solubility experiment of Effect Example 2

[0605] 1) Experimental method

[0606] Take 1.5 mL of brown EP tube, respectively, take 1 mg or so of compound, add 1 mL of ultrapure water. Vortex, ultrasonic to compound no longer dissolved (solution turbid or with suspended particles). Put into incubation oscillator, temperature (37±1℃), 100 r / min, shake 24 h, so as to reach full dissolution balance. After 24 h, take supernatant quickly filter with 0.45 μm microporous filter membrane, discard the primary filtrate, take 200 μL of the filtrate, add 200 μL of methanol dilution. Repeat determination at least three times. According to the chromatographic conditions by LC-MS injection, determine the peak area and calculate the equilibrium solubility of each 2,5-diketopiperazine-imidazole compound in pure water.

[0607] 2) Experimental results

[0608] The solubility experiment results of the compounds are shown in Table 2 as follows:

[0609] Table 2

[0610]

[0611]

[0612] Effect Example 3 Immunofluorescence Experiment

[0613] 1) Experimental method

[0614] 1. Preparation of cell climbing sheet: In the clean bench, sterile round cover glass was placed in a twelve-well plate, NPI-H460 cells in logarithmic growth phase were trypsinized, centrifuged to collect the cells and resuspended with fresh culture medium, 1×10 5 cells were added to each well containing sterile cover glass, after the cells adhered (about 24 h), Plinabulin (10 nM), PLN-3-28 (2 nM) and PLN-3-41 (2 nM) were added and cultured for 24 h.

[0615] 2. Fixation and blocking of cell climbing sheet: the cell climbing sheet was immersed in PBS for 3 min each time, fixed with 4% paraformaldehyde for 15 min, the glass was immersed in PBS for 3 min each time, 0.5% Triton X-100 prepared with PBS was added and allowed to stand for 20 min, the glass was immersed in PBS for 3 min each time, the PBS was discarded, 5% normal goat serum was added to the glass and the glass was blocked at room temperature.

[0616] 3. Addition of primary antibody: the blocking solution was absorbed with water paper, and sufficient primary antibody (prepared with PBS at a certain ratio) was added to the glass, which was placed in a wet box and incubated at 4℃ overnight.

[0617] 4. Add the secondary fluorescent antibody: wash the slide in PBS for 3 times, 3 min each time; after discarding the PBS, add the diluted secondary fluorescent antibody, incubate in a wet box at 37°C for 1 h, and wash the slide in PBS for 3 times, 3 min each time. (From the addition of the secondary fluorescent antibody, each step needs to be operated in the dark)

[0618] 5. DAPI re-stain the cell nucleus: place the slide in PBS (pH 7.4) on a decoloring shaker and shake for 3 times, 5 min each time. After slightly shaking off the slide, add the DAPI staining solution in the circle, and incubate at room temperature for 10 min in the dark.

[0619] 6. Mounting: place the slide in PBS (pH 7.4) on a decoloring shaker and shake for 3 times, 5 min each time. After absorbing the slide with an absorbent paper, mount the slide with an anti-fluorescence quenching mounting agent.

[0620] 7. Microscopy and photographing: observe the slide under a fluorescence microscope and collect images. (The DAPI ultraviolet excitation wavelength is 330-380 nm, the emission wavelength is 420 nm, and blue light is emitted; the FITC excitation wavelength is 465-495 nm, the emission wavelength is 515-555 nm, and green light is emitted; the CY3 excitation wavelength is 510-560, and the emission wavelength is 590 nm, and red light is emitted.)

[0621] 2) Experimental results

[0622] The IOD values were calculated using the software Image Pro Plus 6.0, and the results are shown in Table 3 and Figure 1

[0623] Table 3

[0624]

[0625] The experiment shows that in the blank control group, the untreated cells have normal morphology, complete microtubule skeleton, and regular network filaments. After being treated with the compounds for 24 h, the microtubules appear to be atrophied or even broken. And by comparing the β-tubulin fluorescence values, the microtubule effects of compounds PLN-3-28 and PLN-3-41 are better than Plinabulin.

[0626] Effect implementation example 4 Western Blot experiment

[0627] 1) Experimental method

[0628] (1) Extraction of total cell protein

[0629] The NCI-H460 cells in the logarithmic growth phase were diluted to 1×10 6 ​The density of 1 cell / hole was inoculated in a 6-well plate, and after adherent culture of the cells for 24 h, plinabulin (10 nM), PLN-3-41 (5 nM), and PLN-3-41 (10 nM) were added, respectively, and the plate was incubated in an incubator for 24 h. The culture solution was removed, and the cells were washed twice with PBS. RIPA lysis solution containing protease and phosphatase inhibitors was added to lyse the cells on ice for 30 min. The cells were collected in a 1.5 ml EP tube, centrifuged at 4°C and 12000 rpm for 10 min, and the supernatant was used as the total protein of the cells.

[0630] (2) BCA method for determining protein concentration

[0631] A BSA stock solution with a concentration of 2 mg / ml was diluted with distilled water to 2, 1, 0.5, 0.25, 0.125, and 0.0625 mg / ml. A blank control (distilled water) was also prepared. The protein solution after lysis and centrifugation was diluted with distilled water at a ratio of 1:8. The protein content in a series of BSA solutions with different concentrations and sample protein solutions was determined according to the following method:

[0632] First, 25 μl of solution A (prepared by mixing solution S and solution A at a ratio of 1:50) was added to each sample tube, followed by 5 μl of BSA solution or protein solution with different concentrations. After mixing, 200 μl of solution B was quickly added, and the mixture was incubated at room temperature for 15 min in the dark. The absorbance value was read at a wavelength of 562 nm using a spectrophotometer. The absorbance value of the BSA solution obtained by diluting the standard was used to draw a regression curve. The protein concentration of the sample was calculated by substituting the OD value of the sample into the equation.

[0633] (3). Glue filling

[0634] ① The glue filling glass plate was washed with distilled water and dried vertically.

[0635] ② 10 ml of 15% separation glue was prepared according to the above method, 10 μl of TEMED and 100 μl of 10% ammonium persulfate were added, and the mixture was mixed immediately and filled with glue. The glue was filled to 2-3 mm below the lower edge of the comb, and isopropyl alcohol was used to seal the liquid surface to remove bubbles and isolate air. The mixture was incubated at room temperature for 45 minutes until the separation glue completely solidified.

[0636] ③ After the separation glue completely solidified, the isopropyl alcohol on top was poured off, and the mixture was washed with deionized water three times and dried with filter paper.

[0637] ④ 5 ml of 5% concentrated glue was prepared, 5 μl of TEMED and 50 μl of 10% APS were added, and the mixture was mixed immediately and filled with glue. The glue was filled to the top and a Teflon comb was inserted vertically. The mixture was incubated at room temperature for 20 minutes until the concentrated glue solidified.

[0638] 5. After the gel is completely solidified, remove the comb and place the gel in the electrophoresis tank. Add the running buffer and rinse the wells with the running buffer to remove air bubbles.

[0639] (4). Electrophoresis

[0640] 1. Adjust the protein concentration of the protein extract from each treatment group and mix with an equal volume of 1x loading buffer to obtain the loading solution.

[0641] 2. Boil the loading solution in boiling water at 100°C for 5 min to denature the proteins. Then, chill on ice and centrifuge at 3000 rpm for 1 min.

[0642] 3. Add 50 μg of the loading solution to each well and add 5 μl of pre-stained Marker to one well. Add the running buffer to the wells and cover the tank with the tank lid. Connect the power supply and run at a constant voltage of 80 V for about 15 min. Then, change to a constant voltage of 120 V when the bromophenol blue reaches the separation gel. Turn off the power supply when the bromophenol blue reaches about 0.5 cm from the bottom of the gel. Then, remove the gel plate.

[0643] (5). Protein transfer and immunodetection

[0644] 1. Before the end of the electrophoresis, immerse the PVDF membrane in methanol for 15 s. Then, rinse with transfer buffer for 2 min. After standing for 5 min, start the subsequent operation.

[0645] 2. Pry the gel from the water and soak the gel in the transfer buffer for 15 min after trimming the gel.

[0646] 3. Prepare the transfer "sandwich" in the order of black side (negative) → sponge → filter paper → gel → PVDF membrane → filter paper → sponge → red side (positive). Chase away the air bubbles before adding the next layer. The generation of air bubbles can be avoided by preparing the sandwich in the transfer buffer.

[0647] 4. Connect the positive and negative electrodes and place the transfer cassette in the electroblotter with the PVDF membrane facing the positive electrode. Add the transfer buffer.

[0648] 5. Place the electroblotter in ice water and run at a constant current of 100 mA for 90 min.

[0649] 6. After the transfer is completed, quickly remove the PVDF membrane and seal it in 5% BSA at room temperature for 2 h.

[0650] 7. Remove the membrane and wash it with TBST on a shaker for 5 min x 3 times.

[0651] 8. Add TBST-diluted caspase-3 (dilute the antibody according to the manufacturer's instructions) and GAPDH (dilute the antibody according to the manufacturer's instructions) to the incubation bag. Incubate overnight at 4°C.

[0652] (9) Wash the membrane with TBST for 5 min x 3 times, incubate with HRP-labeled goat anti-rabbit secondary antibody (Biotech brand, 1:7000) or HRP-labeled goat anti-mouse secondary antibody (Biotech brand, 1:7000) at room temperature for 2 h.

[0653] (10) Wash the membrane with TBST for 5 min x 3 times. React the membrane with chemiluminescence detection reagent (reagent A: reagent B = 1:1) for 2 min, and expose and develop in a chemiluminescence developing instrument.

[0654] 2) Experimental results

[0655] The results are shown in Table 4 below:

[0656] Table 4

[0657]

[0658] Experimental results: After 24 h of sample treatment, the relative gray value of Caspase-3 / GAPDH protein was compared, and compared with Plinabulin, PLN-3-41 had a stronger effect on promoting apoptosis of NCI-H460 cell line.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: ； in, R 1 is hydrogen, deuterium, halogen, benzoyl substituted by one or more halogen or "phenoxy substituted by one or more halogen"; R 1a , R 1b , and R 1c are independently hydrogen, deuterium, or halogen; R 2 is hydrogen, deuterium, Ci-C4alkyl substituted by one or more halogens, Ci-C4alkyl or C3-C6cycloalkyl; L stands for "a C1-C5 heteroalkylene with one heteroatom (O)" or "a C1-C5 alkylene". Ring A is C6-C 10 aryl, with one or more R A-1 Replacement C6-C 10 Aryl, "5-10 membered heteroaryl with N as the heteroatom and 1 heteroatom", C3-C6 cycloalkyl or "5-6 membered heterocycloalkyl with N as the heteroatom and 1 heteroatom"; R A-1 independently halogen, cyano, nitro, C1-C4alkyl, C1-C4alkyl substituted by one or more halogens, C1-C4alkoxy, C1-C4alkoxy substituted by one or more halogens, phenyl, or -C(=0)-R 3-1-2 ; R 3-1-2 is C1-C4alkoxy or phenyl.

2. The compound as shown in formula (I) or pharmaceutically acceptable salt thereof according to claim 1, wherein, The compound of formula (I) or its pharmaceutically acceptable salt thereof satisfies one or more of the following conditions: (1) when R 1 is halogen, the halogen is fluorine; (2) when R 1 is benzoyl substituted by one or more halogen, the halogen is fluorine; (3) When R 1 When the phenoxy group is substituted with one or more halogens, the halogen is fluorine; (4) when R 1a , R 1b , and R 1c are independently halogen, said halogen is fluorine; (5) when ring A is C6-C10aryl or "C6-C10aryl substituted by one or more R 10 A-1 substituted C6-C10aryl, said C6-C10aryl is phenyl or naphthyl; 10 substituted C6-C10aryl, said C6-C10aryl is phenyl or naphthyl; 10 substituted C6-C10aryl, said C6-C10aryl is phenyl or naphthyl;​ (6) when R A-1 when independently halogen, said halogen is fluorine, chlorine or bromine.

3. The compound as shown in formula (I) or pharmaceutically acceptable salt thereof according to claim 1, wherein, The compound of formula (I) or its pharmaceutically acceptable salt thereof satisfies one or more of the following conditions: (1) when R 1 is phenyl substituted with one or more halogen, said phenyl substituted with one or more halogen is ; (2) when R 1 is phenoxy substituted by one or more halogen, said phenoxy substituted by one or more halogen is ; (3) When R 2 When the alkyl group is C1-C4, the C1-C4 alkyl group is methyl, ethyl, isopropyl, or tert-butyl; (4) when R is C3-C6cycloalkyl, said C3-C6cycloalkyl is cyclopropyl; 2 (4) when R is C3-C6cycloalkyl, said C3-C6cycloalkyl is cyclopropyl; (5) when L is "C1-C5 heteroalkylene with 1 heteroatom(s) being O", said "C1-C5 heteroalkylene with 1 heteroatom(s) being O" is , or ; (6) when L is C1-C5alkylene, the C1-C5alkylene is methylene, or ; (7) When ring A is C6-C 10 aryl or "by one or more R A-1 Replacement C6-C 10 When "aryl", the C6-C 10 The aryl group is phenyl; (8) When ring A is a "5-10 membered heteroaryl group with N heteroatom and 1 heteroatom", the "5-10 membered heteroaryl group with N heteroatom and 1 heteroatom" is pyridyl, indolyl, indololinyl, pyrroleyl or tetrahydroquinolinyl. (9) When ring A is a C3-C6 cycloalkyl group, the C3-C6 cycloalkyl group is a cyclopentyl group; (10) When ring A is "a 5-6 membered heterocyclic alkyl group with N heteroatom and 1 heteroatom", the "5-6 membered heterocyclic alkyl group with N heteroatom and 1 heteroatom" is pyrrolidinyl or piperidinyl; (11) when R A-1 independently C1-C4alkyl, said C1-C4alkyl is methyl; (12) when R A-1 independently is C1-C4alkyl substituted with one or more halogen, said C1-C4alkyl substituted with one or more halogen is trifluoromethyl; (13) when R A-1 independently C1-C4alkoxy, said C1-C4alkoxy is methoxy; (14) when R A-1 independently is C1-C4alkoxy substituted with one or more halogen, said C1-C4alkoxy substituted with one or more halogen is trifluoromethoxy; (15) when R is C1-C4alkoxy, said C1-C4alkoxy is methoxy. 3-1-2 when R is C1-C4alkoxy, said C1-C4alkoxy is methoxy.

4. The compound as shown in formula (I) or pharmaceutically acceptable salt thereof according to claim 1, wherein, The compound of formula (I) or its pharmaceutically acceptable salt thereof satisfies one or two of the following conditions: (1) when ring A is "5-10 membered heteroaryl having 1 heteroatom which is N", said "5-10 membered heteroaryl having 1 heteroatom which is N" is , , , or ; (2) when ring A is "5-6 membered heterocycloalkyl having 1 heteroatom which is N", said "5-6 membered heterocycloalkyl having 1 heteroatom which is N" is or .

5. The compound as shown in formula (I) or pharmaceutically acceptable salt thereof according to claim 1, wherein, The compound of formula (I) or its pharmaceutically acceptable salt thereof satisfies one or more of the following conditions: (1) R 1 is hydrogen, halogen, benzoyl substituted by one or more halogen or "phenoxy substituted by one or more halogen"; (2) R 1a is hydrogen or halogen; (3) R 1b is hydrogen; (4) R 1c It is hydrogen or halogen; (5) R 2 is hydrogen, C1-C4alkyl substituted by one or more halogen, C1-C4alkyl or C3-C6cycloalkyl.

6. The compound as shown in formula (I) or pharmaceutically acceptable salt thereof according to claim 5, wherein, The compound of formula (I) or its pharmaceutically acceptable salt thereof satisfies one or more of the following conditions: (1) R 1 is hydrogen, F, or ; (2) R 2 Ci-C4-alkyl, C3-C6-cycloalkyl, or C3-C6-cycloalkyl substituted by one or more halogen; (3) L is a C1-C5 alkylene group; (4) Ring A is C6-C 10 aryl, with one or more R A-1 Replacement C6-C 10 Aryl or "5-6 membered heteroaryl with N heteroatom and 1 heteroatom"; (5) R 3-1-2 is C1-C4alkoxy.

7. The compound as shown in formula (I) or pharmaceutically acceptable salt thereof according to claim 1, wherein The compound of formula (I) or its pharmaceutically acceptable salt thereof satisfies one or more of the following conditions: (1) To , , , or ; (2) Ring A is , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

8. The compound as shown in formula (I) or pharmaceutically acceptable salt thereof according to claim 1, wherein, The compound represented by formula (I) is any one of the following compounds: ； ； ； ； ； ； 。 9. A process for the preparation of a compound of formula (I) as claimed in any one of claims 1 to 8, wherein, It includes the following steps: The compound shown in formula (II) undergoes a condensation reaction with the compound shown in formula (III) as shown below to obtain the compound shown in formula (I); ; wherein L, R 1 , R 1a , R 1b , R 1c , R 2 and ring A are as defined in any one of claims 1 to 8.

10. A pharmaceutical composition comprising a compound of formula (I) as described in any one of claims 1-8 or a pharmaceutically acceptable salt thereof, and a pharmaceutical excipient.

11. The use of a compound of formula (I) as claimed in any one of claims 1-8 or a pharmaceutically acceptable salt thereof, or the use of a pharmaceutical composition as claimed in claim 10 in the preparation of a medicament for the prevention and / or treatment of cancer.

12. The use according to claim 11, wherein the compound is ###00011### or a pharmaceutically acceptable salt thereof. The cancer is one or more of the following: lung cancer, pancreatic cancer, colon cancer, and liver cancer.

13. The use of a compound of formula (I) as described in any one of claims 1-8 or a pharmaceutically acceptable salt thereof, or the use of a pharmaceutical composition as described in claim 10, in the preparation of a microtubule inhibitor.

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