Diketopiperazine compound, synthetic method, pharmaceutical composition and application

Abstract

The invention belongs to the technical field of pharmaceutical chemistry, and discloses a diketopiperazine compound, a synthesis method, a pharmaceutical composition and application. The synthesis method comprises the following steps: synthesizing or providing imidazole formaldehyde C; synthesizing or providing an intermediate 2, 5-diketopiperazine derivative B; synthesizing or providing 3-substituted benzaldehyde A; 3-substituted benzaldehyde A and an intermediate 2, 5-diketopiperazine derivative B are subjected to Aldol condensation to obtain the diketopiperazine compound with the structure as shown in the formula (I). The invention also discloses application of the compound in preparation of medicines for preventing or treating cancers, especially application in preparation of medicines for treating lung cancers. The compound disclosed by the invention is novel in structure, part of the compound has excellent anti-tumor activity, all the compounds have excellent solubility, and the compound has a good development prospect.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical chemistry, and particularly relates to a diketopiperazine compound, a synthetic method, a pharmaceutical composition and a use. BACKGROUND

[0002] Phenylahistin is a natural product isolated from the marine fungus Aspergillus ustus, which acts on the colchicine site. Its derivative, Plinabulin, has entered the phase III clinical research stage. The structure of the complex of Phenylahistin derivatives combined with tubulin at the colchicine site has been resolved, which shows that the colchicine site has multiple sites that can form molecular forces, directly or through a water bridge to form hydrogen bonds with amino acids. And the binding pocket of colchicine penetrates into the beta subunit, across the alpha / beta interface, and extends to the boundary of the GTP pocket formed by hydrophilic and hydrophobic amino acid residues. Phenylahistin derivatives cause the overall structure of tubulin dimers to bend by affecting the conformation of the T7 loop (amino acid residue number: 244-251) and the H7 helix (amino acid residue number: 224-243), and depolymerize microtubules. Plinabulin has the problems of low efficacy of single drug and high toxicity (possible reasons: poor pharmacokinetic properties, poor physicochemical properties, and poor selectivity). First, the solubility of Plinabulin is poor (<0.1 mg / mL), which cannot be used as an oral preparation for administration; second, the in vivo activity of single drug is still not ideal, and a clinical administration scheme of combined administration with Docetaxel needs to be used, and the experimental dose of the phase III clinical study is 30 mg / m 2 Plinabulin and 75 mg / m 2 Docetaxel; and third, the selectivity is poor, and there is a strong toxic side effect on normal cell tissues, and the treatment window is narrow. Moreover, the above problems have not been solved by the preferred derivatives of Phenylahistin. The prior art Plinabulin compound is as follows:

[0003] SUMMARY

[0004] To overcome the problems in the related art, the present application provides a diketopiperazine compound, a synthetic method, a pharmaceutical composition and a use, and particularly relates to a 1,3-imidazole-2,5-diketopiperazine compound and its application in the preparation of an anticancer drug.

[0005] The technical solution is as follows: a diketopiperazine compound, the general formula (I) of the molecular structure of the diketopiperazine compound is:

[0006]

[0007] wherein each R1is independently selected from a hydrogen atom, an alkyl group, an alkenyl group, an alkynyl group, an aryl group, an alkoxy group, an amido group, or an acyloxy group, or

[0008] R2is selected from any one of a hydrogen atom, an alkyl group, an aryl group, an aromatic heterocycle, a saturated polyheterocycle, and an unsaturated polyheterocycle

[0009] Further, the X is -O- or -C=O-.

[0010] The R1' in para position to the X group is a fluorine atom or a hydrogen atom.

[0011] The R2is one of a methyl group, an ethyl group, an isopropyl group, or a tert-butyl group.

[0012] The R2is selected from

[0013] one of the following.

[0014] Another object of the present application is to provide a pharmaceutical composition comprising a therapeutically effective dose of the diketopiperazine compound and a pharmaceutically acceptable carrier; the IC 50 value of the therapeutically effective dose of the diketopiperazine compound is 1 uM-0.02 uM.

[0015] Another object of the present application is to provide a use of the diketopiperazine compound in the preparation of a medicament for preventing and / or treating cancer.

[0016] Another object of the present application is to provide a use of the pharmaceutical composition in the preparation of a medicament for preventing and / or treating cancer.

[0017] Another object of the present application is to provide a synthesis method of a diketopiperazine compound, for the synthesis of the diketopiperazine compound, the method comprising:

[0018] S1, synthesizing or providing an imidazole aldehyde C;

[0019] S2, synthesizing or providing an intermediate 2,5-diketopiperazine derivative B;

[0020] S3, synthesizing or providing a 3-substituted benzaldehyde A;

[0021] S4, performing Aldol condensation of the 3-substituted benzaldehyde A and the intermediate 2,5-diketopiperazine derivative B to obtain a compound having a structure shown in Formula (I), i.e., a diketopiperazine compound.

[0022] Furthermore, in step S1, K2CO3, DMF, 50℃, 6h, 26.0%-71.58%;

[0023] In step S2, Cs2CO3, Na2SO4, DMF, 45-70℃, 22h-24h, N2 protection, 25.8%-72.0%;

[0024] In step S3, Cu(OAc)2, TEA, Dry DCM, O2, rt, 22.0%;

[0025] In step S3, Cs2CO3, Na2SO4, DMF, rt-50℃, 6h-22h, N2 protection, 45.6%-47.0%.

[0026] Combining all the above technical solutions, the beneficial effects of this invention are as follows:

[0027] This invention discloses a compound having the structure of formula (I), its isomers, or its pharmaceutically acceptable salts. This invention also discloses the use of such compounds in the preparation of medicaments for the prevention or treatment of cancer, particularly in the preparation of medicaments for the treatment of lung cancer. The compounds of this invention have novel structures, and some compounds exhibit excellent antitumor activity and solubility, showing promising development prospects.

[0028] The present invention provides a class of compounds that, while possessing protective activity, have significantly improved solubility through the addition of hydrochloride salts. Solubility is a core indicator for evaluating the physicochemical properties of drug molecules, and this invention represents a significant breakthrough in this area.

[0029] The compounds involved in this invention are novel compounds obtained by the inventors through structural and synthetic route design and chemical synthesis, and have not been reported in the prior art. Some compounds have antitumor activity comparable to or even better than punabulin. In particular, when X is -O- in the compound of formula (I), the water solubility and bioavailability of the compounds of this invention are improved compared to punabulin, and they have good development prospects. Attached Figure Description

[0030] The accompanying drawings, which are incorporated in and form part of this specification, illustrate embodiments consistent with this disclosure and, together with the description, serve to explain the principles of this disclosure;

[0031] Figure 1 This is a flowchart of the method for synthesizing diketopiperazine compounds provided in the embodiments of the present invention. Detailed Implementation

[0032] In order to make the above objectives, features and advantages of the present application more clear and comprehensible, specific embodiments of the present application will be described in detail below with reference to the accompanying drawings. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present application. However, it will be apparent to one skilled in the art that the present application can be practiced without the specific details given herein. In other instances, well-known methods have not been described in detail in order to avoid obscuring the present application. Unless otherwise specified, the scientific and technical terms used in this application should have the meanings commonly understood by one of ordinary skill in the art. However, the following terms are defined below:

[0033] The methods and techniques of the present application are generally performed according to conventional methods known to those of skill in the art unless otherwise specified. The nomenclature used in connection with, and the laboratory procedures and techniques of, biological, pharmacological, and medicinal chemistry described herein are those known and practiced by those in the art. Standard techniques are used for chemical syntheses, chemical analyses, pharmaceutical synthesis, formulation, and delivery of drugs, and treatment of patients.

[0034] Unless otherwise specified, the scientific and technical terms used in this application should have the meanings commonly understood by one of ordinary skill in the art. However, the following terms are defined below:

[0035] The term "isomers" includes conformational isomers, optical isomers (such as enantiomeric and diastereomeric isomers), and geometric isomers (such as cis / trans isomers). These isomers or combinations thereof can exist as racemic mixtures (racemates), as individual enantiomers, as individual diastereomers, as mixtures of diastereomers, as cis or trans isomers.

[0036] The term "pharmaceutically acceptable salts" refers to any pharmaceutically acceptable salt of a compound of formula (I), preferably to an acid addition salt of the compound. Examples of preferred pharmaceutically acceptable salts are acid addition salts of pharmaceutically acceptable inorganic or organic acids, such as for example hydrohalic acids, sulfuric acid, phosphoric acid, or aliphatic or aromatic carboxylic or sulfonic acids, for example acetic acid, succinic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, nicotinic acid, methanesulfonic acid, p-toluenesulfonic acid or naphthalenesulfonic acid. Examples of other preferred pharmaceutically acceptable salts are alkali metal salts (of sodium or potassium), alkaline earth metal salts (of calcium or magnesium), or ammonium salts derived from ammonia or from pharmaceutically acceptable organic amines, such as for example C1-C7-alkylamines, cyclohexylamine, triethanolamine, ethylenediamine or tris(hydroxymethyl)aminomethane.

[0037] The term "halogen atom" refers to any of the radio stable atoms of column 7 of the Periodic Table of the Elements, i.e. fluorine, chlorine, bromine or iodine, preferably fluorine and chlorine.

[0038] The term "alkyl" refers to any straight-chain or branched, substituted or unsubstituted saturated hydrocarbon group, preferably a hydrocarbon group with 1-10 carbon atoms, including but not limited to methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, 2,4,4-trimethylpentyl, cyclopentyl, n-hexyl, isohexyl, cyclohexyl, n-heptyl, cycloheptyl, n-octyl, 2-ethylhexyl, cyclooctyl, n-nonyl, cyclononyl, or n-decyl. Among the substituted saturated hydrocarbon groups, C1-C10 monohalogenated, dihalogenated, and perhalogenated saturated hydrocarbon groups, as well as amino-substituted hydrocarbon groups, are preferred, with perfluoromethyl, perchloroethyl, perfluoroisopropyl, and perchlorotert-butyl being the most preferred.

[0039] The term "acyl" refers to an alkyl or aryl group derived from an oxyacid, preferably an acetyl group.

[0040] The term "alkenyl" refers to any straight-chain or branched, substituted or unsubstituted unsaturated hydrocarbon group, including polyunsaturated hydrocarbon groups, preferably alkenyl groups with 2-10 carbon atoms, including but not limited to vinyl, allyl, methyl vinyl, propenyl, butenyl, pentenyl, 1,1-dimethyl-2-propenyl, hexenyl, cyclopropenyl, cyclobutenyl, cyclopentenyl or cyclohexenyl.

[0041] The term "alkynyl" is preferably an alkynyl group having 2-5 carbon atoms, including but not limited to ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 1-methyl-2-propynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 1-ethyl-2-propynyl, 2-pentynyl, 3-pentynyl, 1-methyl-2-butynyl, 4-pentynyl, 1-methyl-3-butynyl, or 2-methyl-3-butynyl.

[0042] The term "alkoxy" refers to any straight-chain or branched, substituted or unsubstituted, saturated or unsaturated ether, preferably an alkoxy group having 1-10 carbon atoms, including but not limited to methoxy, ethoxy, n-propoxy, isopropoxy, cyclopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, cyclobutoxy or cyclohexyloxy.

[0043] The terms "aryl," "substituted aryl," "heteroaryl," and "substituted heteroaryl" refer to an aromatic hydrocarbon ring, preferably having 5, 6, or 7 atoms, and most preferably 6 atoms constituting the ring. "Heteroaryl" and "substituted heteroaryl" refer to an aromatic hydrocarbon ring having at least one heteroatom (e.g., oxygen, sulfur, or nitrogen atom) and at least one carbon atom. The term "aryl" includes, but is not limited to, substituted or unsubstituted groups of the following: phenyl, o-methylphenyl, m-tolyl, xylyl, naphthyl, or anthracene.

[0044] The term "multi-membered heterocycle" is preferably a substituted or unsubstituted saturated nitrogen-containing multi-membered heterocycle, a substituted or unsubstituted unsaturated sulfur-containing multi-membered heterocycle, or a substituted or unsubstituted unsaturated oxygen-containing multi-membered heterocycle; more preferably, the multi-membered heterocycle is a 4-6 membered heterocycle.

[0045] The term "substituted" refers to a group in which at least one hydrogen atom is replaced by a substituent selected from halogen atoms, alkyl, substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyacylamino, cyano, hydroxy, carboxyl, carboxylalkyl, ketone, thioketone, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclicoxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, and -SO2-heteroaryl.

[0046] The term "therapeutic effective dose" refers to any amount of a drug, when used alone or in combination with another therapeutic agent, that promotes disease regression, manifested as a reduction in the severity of disease symptoms, an increase in the frequency and duration of symptom-free periods, or prevention of impairment or disability caused by the disease. The "therapeutic effective dose" of the drug of this invention also includes a "preventive effective dose," which refers to an amount of the drug, when administered alone or in combination with another therapeutic agent to a subject at risk of developing the disease or experiencing a relapse, that inhibits the occurrence or recurrence of the disease.

[0047] As will be apparent to those skilled in the art, the effective in vivo dosage and specific route of administration will vary depending on the species, weight, and age of the mammal being treated, the specific compound used, and the specific purpose for which the compound is used. Those skilled in the art can determine the effective dosage level (i.e., the dosage level necessary to achieve the desired effect) using conventional pharmacological methods. Typically, the human clinical application of a product begins at a lower dosage level, subsequently increasing the dosage level until the desired effect is achieved. Alternatively, the useful dosage and route of administration of the compositions identified by this method can be established using acceptable in vitro studies employing existing pharmacological methods.

[0048] When used as an anticancer or antitumor agent, the compound is preferably administered continuously in a stable dose experimental animal model, for example, via intravenous infusion. However, those skilled in the art will understand that in certain situations it may be necessary to administer the antitumor compound at doses exceeding or even far exceeding the preferred dose range described above, thereby effectively and aggressively treating specific advanced or lethal tumors.

[0049] The term "pharmaceutically acceptable carrier" includes acceptable excipients, diluents, preservatives, stabilizers, dyes, flavorings, antioxidants, and suspending agents that can be used for medical purposes.

[0050] The "pharmaceutical compositions" of this invention can be formulated or used in the following forms: tablets, capsules or elixirs for oral administration; suppositories for rectal administration; sterile solutions or suspensions for injection; patches for transdermal administration; and subcutaneous deposits, etc. Injectable formulations can be formulated in conventional forms, such as liquid solutions or suspensions, solid forms suitable for dissolving or suspending in a liquid prior to injection or infusion, or emulsions.

[0051] The term "cancer" refers to a broad class of diseases characterized by the uncontrolled growth of abnormal cells within the body. Uncontrolled cell division and growth lead to the formation of malignant tumors or cells that invade adjacent tissues and can also metastasize to distant parts of the body via the lymphatic system or bloodstream. In this invention, another equivalent description of "treating cancer" is "treating tumors," "anti-cancer," or "antitumor."

[0052] Cancer is an uncontrolled growth of cells that impairs the normal function of the body's organs and systems. A subject with cancer is one with objectively measurable amounts of cancer cells present in their body. Subjects at risk of developing cancer are those who are predisposed to cancer (e.g., based on family history or genetic predisposition), or those exposed to radiation or other carcinogenic agents.

[0053] The compounds and pharmaceutical compositions thereof of the present invention can be used to prevent or treat a variety of cancers. Examples of such cancers include breast cancer, prostate cancer, lung cancer, ovarian cancer, cervical cancer, skin cancer, melanoma, colon cancer, stomach cancer, liver cancer, esophageal cancer, kidney cancer, pharyngeal cancer, thyroid cancer, pancreatic cancer, testicular cancer, brain cancer, bone cancer, and blood cancers (such as leukemia, chronic lymphocytic leukemia), etc. Other cancers include, but are not limited to, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain and central nervous system (CNS) cancer, cervical cancer, choriocarcinoma, colorectal cancer, connective tissue cancer, digestive system cancer, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, intraepithelial neoplasia, laryngeal cancer, lung cancer (small cell, large cell), lymphoma (including Hodgkin's lymphoma and non-Hodgkin's lymphoma), melanoma, neuroblastoma, oral cancer (e.g., lip, tongue, mouth and pharynx), retinoblastoma, rhabdomyosarcoma, respiratory system cancers, sarcomas, uterine cancer, urinary system cancers, and other cancers and sarcomas. The compounds and compositions thereof of the present invention are preferably used for lung cancer.

[0054] The innovative aspects of the diketopiperazine compounds, synthesis methods, pharmaceutical compositions, and uses provided in the embodiments of this invention are as follows:

[0055] This invention targets the colchicine site of tubulin, a proven important anti-tumor target, yet no anti-tumor drugs have been marketed there to date. Based on the three-dimensional spatial structure of the target and ligand, a new space for modifying the imidazole ring was discovered. By comprehensively applying structure-based drug design and classical drug design strategies, the structure of the lead compound was modified in regions with less space. While evaluating efficacy, physicochemical properties were tested, providing crucial support for improving the druggability of this class of compounds. The prepared library of phenylahistin derivatives, a tubulin inhibitor, exhibits novel structures and systematic structure-activity relationship studies, offering possibilities for the discovery of original anti-tumor drugs. Through detailed studies of the relationship between compound structure, activity, and water solubility, structure-activity and structure-solution curves were established. The results show that these compounds maintain good activity while significantly improving solubility, laying the foundation for discovering drug candidates with good activity, pharmacokinetic stability, and high bioavailability.

[0056] Example: A diketopiperazine compound, as shown in general formula (I):

[0057]

[0058] Its isomers or pharmaceutically acceptable salts include:

[0059] Another exemplary embodiment of the present invention provides a salt formation according to the compound described above. The salt solubility of the diketopiperazine compound is greater than 1.00 mg / ml.

[0060] Another exemplary embodiment of the present invention provides a hydrochloride salt of the compound with a solubility range of 1.14 mg / ml to 354.47 mg / ml.

[0061] R1 is independently selected from hydrogen atom, alkyl, alkenyl, alkynyl, aryl, alkoxy, amide, or acyloxy group or

[0062] R2 is selected from any one of hydrogen atoms, alkyl groups, aryl groups, aromatic heterocycles, saturated multi-heterocycles, and unsaturated multi-heterocycles.

[0063] For example, X is -O- or -C=O-.

[0064] For example, R1', which is in the para position to the X group, is a fluorine atom or a hydrogen atom.

[0065] For example, R2 is one of methyl, ethyl, isopropyl or tert-butyl.

[0066] Another example is that R2 is selected from...

[0067] One of them.

[0068] Another exemplary embodiment of the present invention provides a pharmaceutical composition comprising:

[0069] A therapeutically effective dose of the diketopiperazine compound, and a pharmaceutically acceptable carrier. The therapeutically effective dose of the diketopiperazine compound at IC50... 50 The value is 1uM-0.028uM.

[0070] Another exemplary embodiment of the present invention provides the use of the said compound or the said pharmaceutical composition in the preparation of a medicament for the prevention and / or treatment of cancer.

[0071] An example of the cancer mentioned is lung cancer.

[0072] like Figure 1 As shown, the synthetic methods for diketopiperazine compounds include:

[0073] S1, synthesizes or provides imidazole formaldehyde C;

[0074] S2, synthesizes or provides intermediate 2,5-dikepiperazine derivative B;

[0075] S3, synthesizing or providing 3-position substituted benzaldehyde A;

[0076] S4, 3-substituted benzaldehyde A is condensed with intermediate 2,5-diketopiperazine derivative B by Aldol to obtain a compound having the structure shown in formula (I), namely a diketopiperazine compound.

[0077] For example, the synthetic parameters involved in each step of the synthesis method for diketopiperazine compounds include:

[0078] In step S1, K2CO3, DMF, 50℃, 6h, 26.0%-71.58%;

[0079] In step S2, Cs2CO3, Na2SO4, DMF, 45-70℃, 22h-24h, N2 protection, 25.8%-72.0%;

[0080] In step S3, Cu(OAc)2, TEA, Dry DCM, O2, rt, 22.0%;

[0081] In step S3, Cs2CO3, Na2SO4, DMF, rt-50℃, 6h-22h, N2 protection, 45.6%-47.0%.

[0082] As can be seen from the above embodiments, the present invention obtains a class of innovative drug candidate compounds, which can then be transformed into clinical trials and enter the market, showing good economic prospects; it contributes to the high-value utilization of marine biological resources, and obtains innovative drug candidate compounds of diketopiperazine based on the structure of marine natural products; subsequent systematic pharmacological and toxicological studies of the candidate compounds can be carried out to evaluate their potential clinical application value in treating tumors, which will ultimately generate huge economic benefits.

[0083] This invention establishes an efficient method for preparing 2,5-piperazine dione compounds, expanding the core technical means for preparing these compounds and providing a material basis for further exploring their potential medicinal value. Furthermore, the implementation of this patented invention has deeply explored the functions of different types of pharmacophores, significantly improving the physicochemical properties of colchicine site compounds, providing a foundation for oral administration of these substances, significantly enhancing the drug-likeness of marine natural products, and highlighting the important social value of marine resources in meeting clinical needs.

[0084] This invention is guided by the research and development of innovative drug products. Through the implementation of this patent, the problem of insufficient water solubility of this type of compound has been solved, laying the foundation for further clinical development of this type of small molecule.

[0085] This invention establishes an optimized technology system and an efficient preparation technology system for marine natural products, laying the foundation for the research and development of innovative marine small molecule drugs.

[0086] To further illustrate the effects of the embodiments of the present invention, the following experimental examples were conducted. Examples of some diketopiperazine compounds of the present invention are shown in Table 1.

[0087] Table 1 Examples of diketopiperazine compounds

[0088]

[0089]

[0090]

[0091] The technical solution of the present invention will be further described in detail below with reference to specific embodiments.

[0092] Example 1: Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-0-1).

[0093] (1) Synthesis of (Z)-1-acetyl-3-benzylidenepiperazine-2,5-dione (BI-01):

[0094]

[0095] Benzaldehyde (1 g, 9.42 mmol), 1,4-diacetylpiperazin-2,5-dione (3.73 g, 18.8 mmol), cesium carbonate (4.1 g, 12.58 mmol), and DMF (30 mL) were weighed into a dry 100 mL single-necked flask. The mixture was purged with nitrogen and reacted at room temperature for 6 h. The reaction solution was poured into cold water, resulting in the precipitation of a large amount of solid. After filtration and drying, 2.67 g of a white solid was obtained and used directly in the next reaction step. 1 H NMR(500MHz,DMSO-d6)δ9.96(s,1H),7.67(dt,J=7.6,1.3Hz,1H),7.61(t,J=7.8Hz ,1H),7.39–7.33(m,2H),7.30–7.25(m,2H),7.17–7.13(m,2H).MS(ESI)m / z:[M+H] + Calcd for C 13 H 10 FO2:217.0659, Found216.01.

[0096] (2) Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1H-imidazol-2-yl)methylene)piperazine-2,5-dione:

[0097]

[0098] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), imidazole-2-carboxaldehyde (88.5 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (142.04 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place under nitrogen protection, and in an oil bath at 70 °C. Stir and react for 24 h. After the reaction is complete, drop the reaction solution into water (50 mL), let stand for 2 h, filter, and dry. Sonicate the solution with ethyl acetate, let stand in a refrigerator at 4 °C for 2 h, filter, and obtain 66 mg of yellow solid, yield 38.40%. Melting point: 289-291 °C. 1 HNMR(500MHz,DMSO-d6)δ12.01(s,1H),7.51(d,J=7.3Hz,2H),7.39(dd,J=8.4,6.8Hz,2H), 7.31(dd,J=14.9,7.7Hz,2H),7.22(s,1H),6.79(s,1H),6.52(s,1H),2.46(p,J=1.9Hz,1H). 13 C NMR(125MHz,DMSO-d6)δ157.26,157.20,144.06,133.46,129.93,129.90,129.17,128.74,128.60,126.64,98.91.MS(ESI)m / z:[M+H] + Calcd for C 15 H 13 N4O2:281.1033, Found281.1031.

[0099] Example 2 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-methyl-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-0-2).

[0100]

[0101] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), 1-methyl-1H-imidazolium-2-carboxaldehyde (101.4 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (142.04 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 50 mL of water, let stand for 2 h, filter, and dry. Slurry the mixture with ethyl acetate, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 112 mg of yellow solid, yield 59.82%. Melting point: 276-277 °C. 1 H NMR(500MHz,DMSO-d6)δ12.23(d,J=9.6Hz,1H),10.37(s,1H),7.58–7.50(m,2H),7.42(dt,J=7.4,5.1H z,2H),7.38–7.29(m,2H),7.24–7.19(m,1H),6.85–6.80(m,1H),6.60–6.55(m,1H),3.81–3.73(m,3H). 13 C NMR (125MHz, DMSO-d6) δ157.32,157.19,143.99,133.40,129.93,129.16,128. 98,128.77,128.74,126.55,123.47,115.87,96.26,32.97.MS(ESI)m / z:[M+H] + Calcd for C 16 H 15 N4O2: 295.1190, Found 295.1189.

[0102] Example 3 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-ethyl-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-0-3).

[0103] 1) Preparation of 1-ethyl-1H-imidazol-2-carboxaldehyde (BI-02):

[0104]

[0105] Take a 25 mL dry round-bottom flask and add imidazole-2-carboxaldehyde (200 mg, 2.08 mmol), DMF (3 mL), potassium carbonate (345.2 mg, 2.50 mmol), and iodoethane (389.6 mg, 2.50 mmol) in DMF sequentially. Place the flask in an oil bath at 50 °C and stir for 6 h. Filter, dilute with water, extract with EA, wash the organic phase with saturated brine, dry to anhydrous sodium sulfate, and concentrate under reduced pressure to give 180.9 mg of a brown oily substance, yield 70%. 1 H NMR (400MHz, CDCl3) δ9.81 (s, 1H), 7.28 (s, 1H), 7.17 (s, 1H), 4.43 (q, J = 7.3Hz, 2H), 1.43 (t, J = 7.3Hz, 3H). MS (ESI) m / z: [M+H] + Calcd for C6H9N2O:125.0709,Found:124.69

[0106] 2) Synthesis of the target product:

[0107]

[0108] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), 1-ethyl-1H-imidazolium-2-carboxaldehyde (114 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 50 mL of water, let stand for 2 h, filter, and dry. Sonicate the solution with ethyl acetate, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 135 mg of yellow solid, yield 71.43%. Melting point: 208-209 °C. 1 H NMR (500MHz, DMSO-d6) δ12.28(d,J=9.8Hz,1H),10.37(d,J=9.5Hz,1H),7.58–7.49(m,2H),7.47–7.37(m,3H),7.37–7.28(m ,1H),7.27–7.21(m,1H),6.85–6.79(m,1H),6.60–6.55(m,1H),4.16(dd,J=11.8,7.2Hz,2H),1.31(dt,J=11.5,7.2Hz,3H). 13C NMR(125MHz,DMSO-d6)δ157.32,157.22,143.07,133.41,129.94,129.19,129. 16,129.02,128.77,126.55,121.78,115.89,96.04,16.88.MS(ESI)m / z:[M+H] + Calcd for C 17 H 17 N4O2: 309.1346, Found 309.1348.

[0109] Example 4 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-propyl-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-0-4).

[0110] 1) Synthesis of 1-propyl-1H-imidazol-2-carboxaldehyde (BI-03):

[0111]

[0112] The compound BI-02 was prepared according to the synthetic method described above. Using imidazole-2-carboxaldehyde (400 mg, 4.16 mmol), potassium carbonate (690.4 mg, 5.00 mmol), and 1-iodopropane (849.2 mg, 5.00 mmol) as starting materials, and DMF (10 mL), the reaction was carried out in an oil bath at 50 °C with stirring for 6 h. 373.9 mg of an oily liquid was obtained, with a yield of 65%. 1 H NMR(500MHz, CDCl3)δ9.85(s,1H),7.31(s,1H),7.15(s,1H),4.39–4.33(m,2H) ,1.82(dd,J=14.6,7.3Hz,2H),0.94(dt,J=13.4,7.4Hz,5H).MS(ESI)m / z:[M+H] + Calcd for C7H 11 N2O: 139.0866, Found: 138.64.

[0113] 2) Synthesis of the target product:

[0114]

[0115] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), 1-propyl-1H-imidazolium-2-carboxaldehyde (127.3 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 50 mL of water, let stand for 2 h, filter, and dry. Slurry the mixture with ethyl acetate using ultrasound, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 51 mg of a yellow solid, yield 25.88%. Melting point: 186-187 °C. 1 H NMR (500MHz, DMSO-d6) δ12.36–12.30(m,1H),10.39(s,1H),7.56(dd,J=7.7,1.9Hz,2H),7.47–7.40(m,3H),7.35(dd,J=7.3,1.9Hz,1H),7.26(d, J=1.6Hz,1H),6.84(d,J=1.9Hz,1H),6.60(d,J=1.9Hz,1H),4.12(td,J=7.2,1.9Hz,2H),1.72(qd,J=7.2,1.9Hz,2H),0.86(td,J=7.4,1.9Hz,3H). 13 C NMR(125MHz,DMSO-d6)δ157.34,157.24,143.43,133.43,129.94,129.21,129.15,128 .88,128.75,126.60,122.47,115.86,96.10,47.23,24.51,11.29.MS(ESI)m / z:[M+H] + Calcd for C 18 H 19 N4O2: 323.1503, Found 323.1507.

[0116] Example 5 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-isopropyl-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-0-5).

[0117] 1) Synthesis of 1-isopropyl-1H-imidazol-2-carboxaldehyde (BI-04):

[0118]

[0119] The compound BI-02 was prepared according to the synthetic method described above. Using imidazole-2-carboxaldehyde (400 mg, 4.16 mmol), potassium carbonate (690.4 mg, 5.00 mmol), and 2-iodopropane (849.2 mg, 5.00 mmol) as starting materials, and DMF (10 mL), under nitrogen protection, the reaction was carried out in an oil bath at 50 °C with stirring for 6 h. 74.0 mg of an oily liquid was obtained, with a yield of 61%. 1 H NMR (500MHz, CDCl3) δ9.82 (s, 1H), 7.32 (s, 1H), 7.30 (s, 1H), 5.46 (dt, J = 13.4, 6.7Hz, 1H), 1.46 (d, J = 6.7Hz, 6H). MS (ESI) m / z: [M+H] + Calcd for C7H 11 N2O: 139.0866, Found: 139.04.

[0120] 2) Synthesis of the target product:

[0121]

[0122] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (200 mg, 0.82 mmol), 1-isopropyl-1H-imidazolium-2-carboxaldehyde (170 mg, 1.23 mmol), cesium carbonate (400.2 mg, 1.22 mmol), anhydrous sodium sulfate (233 mg, 1.64 mmol), and DMF (10 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 50 mL of water, let stand for 2 h, filter, and dry. Slurry the mixture with ethyl acetate, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 163 mg of yellow solid, yield 61.74%. Melting point: 196-198 °C. 1 H NMR (500MHz, DMSO-d6) δ12.36(s,1H),10.38(s,1H),7.56(dt,J=6.1,1.5Hz,3H),7.43(t,J=7.6Hz,2H),7.35( d,J=7.1Hz,1H),7.28(d,J=1.3Hz,1H),6.83(s,1H),6.64(s,1H),3.35(d,J=1.0Hz,1H),1.41(d,J=6.6Hz,6H). 13C NMR(125MHz,DMSO-d6)δ157.31(d,J=7.7Hz),129.94,129.30,129.16,128.76,118.47,115.86,96.09,47.54,23.63.MS(ESI)m / z:[M+H] + Calcd for C 18 H 19 N4O2: 323.1503, Found 323.1506.

[0123] Example 63: Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-benzimidazole-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-0-6).

[0124]

[0125] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), benzimidazole-2-carboxaldehyde (134.6 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place under nitrogen protection, and in an oil bath at 70 °C. Stir and react for 24 h. After the reaction is complete, drop the reaction solution into water (50 mL), let stand for 2 h, filter, and dry. Slurry the mixture with ethyl acetate, place in a refrigerator at 4 °C, let stand for 2 h, filter, and obtain 88 mg of yellow solid, yield 43.34%. Melting point: 228-229 °C. 1 H NMR (500MHz, DMSO-d6) δ12.25(s,1H),7.81–7.71(m,1H),7.57(d,J=7.7Hz,3H),7.43(t,J=7.6Hz,2H),7.38–7.24(m,3H),6.90(s,1H),6.65(s,1H). 13 C NMR(125MHz,DMSO-d6)δ157.47,156.72,149.71,133.35,132.96,130.03,129.17,128.89,126.45,116.68,97.69.MS(ESI)m / z:[M+H] + Calcdfor C 19 H 15 N4O2: 331.1190 Found 331.1192.

[0126] Example 7 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-allyl-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-0-9).

[0127] 1) Synthesis of 1-allyl-1H-imidazol-2-carboxaldehyde (BI-05):

[0128]

[0129] The compound BI-02 was prepared according to the synthetic method described above. Using imidazole-2-carboxaldehyde (200 mg, 2.08 mmol), DMF (5 mL), potassium carbonate (345.2 mg, 2.50 mmol), and 3-bromo-propene (302.2 mg, 2.50 mmol) as starting materials, and DMF (5 mL), the reaction was carried out in an oil bath at 50 °C with stirring for 6 h. 158.7 mg of an oily liquid was obtained, with a yield of 56%. 1 H NMR (500MHz, CDCl3) δ9.82 (s, 1H), 7.30 (s, 1H), 7.16 (s, 1H), 5.95 (ddt, J = 16.1, 10.4, 5.8Hz, 1H), 5 .24(dd,J=10.2,0.8Hz,1H),5.10(dd,J=17.0,0.7Hz,1H),5.02(d,J=5.8Hz,2H).MS(ESI)m / z:[M+H] + Calcd forC7H9N2O:137.0709,Found:136.65.

[0130] 2) Synthesis of the target product:

[0131]

[0132] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), 1-allyl-1H-imidazolium-2-carboxaldehyde (124.52 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 50 mL of water, let stand for 2 h, filter, and dry. Slurry the mixture with ethyl acetate using ultrasound, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 119 mg of a yellow solid, yield 60.40%. Melting point: 207-208 °C. 1H NMR (500MHz, DMSO-d6) δ12.25(d,J=2.8Hz,1H),10.37(s,1H),7.55(dd,J=7.9,2.8 Hz,2H),7.41(ddd,J=15.5,7.6,2.5Hz,3H),7.33(dd,J=8.5,5.8Hz,1H),7.28(d,J =2.7Hz,1H),6.83(d,J=2.7Hz,1H),6.53(d,J=2.8Hz,1H),6.07–5.95(m,1H),5.19 (dd, J=9.4, 2.9Hz, 1H), 4.90 (dd, J=17.1, 3.3Hz, 1H), 4.84 (dd, J=5.0, 2.6Hz, 2H). 13 C NMR (125MHz, DMSO-d6) δ157.35,157.16,143.68,134.54,133.42,129.94,129.28,129. 15,129.00,128.76,126.56,122.61,117.21,115.91,96.23,48.03.MS(ESI)m / z:[M+H] + Calcd for C 18 H 17 N4O2: 321.1346, Found 321.1347.

[0133] Example 8 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-benzyl-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-0-10).

[0134] 1) Synthesis of 1-benzyl-1H-imidazol-2-carboxaldehyde (BI-06):

[0135]

[0136] The compound BI-02 was prepared according to the synthetic method described above. Using imidazole-2-carboxaldehyde (200 mg, 2.08 mmol), potassium carbonate (575.3 mg, 4.16 mmol), and benzyl bromide (302.4 mg, 1.87 mmol) as starting materials, the mixture was stirred in an oil bath at 50 °C for 6 h with 10 mL of DMF. Thin-layer chromatography yielded 166.5 mg of an oily liquid, with a yield of 43%. 1 H NMR (500MHz, CDCl3) δ9.86 (s, 1H), 7.37–7.29 (m, 4H), 7.20 (d, J = 6.9Hz, 2H), 7.13 (s, 1H), 5.61 (s, 2H). MS (ESI) m / z: [M+H] +Calcd for C 11 H 11 N2O: 187.0866, Found: 186.72

[0137] 2) Synthesis of the target product:

[0138]

[0139] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), 1-benzyl-1H-imidazolium-2-carboxaldehyde (158.6 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 50 mL of water, let stand for 2 h, filter, and dry. Slurry the mixture with ethyl acetate, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 175 mg of yellow solid, yield 63.18%. Melting point: 233-234 °C. 1 H NMR(500MHz,DMSO-d6)δ12.26(s,1H),10.36(s,1H),7.56–7.51(m,3H),7. 44–7.27(m,7H),7.18–7.12(m,2H),6.82(s,1H),6.60(s,1H),5.46(s,2H). 13 C NMR (125MHz, DMSO-d6) δ157.36,157.09,143.79,137.71,133.43,129.93,129.50,129.30,129. 20,129.14,128.75,128.20,127.11,126.58,123.00,115.92,96.06,48.97.MS(ESI)m / z:[M+H] + Calcd forC 22 H 19 N4O2: 371,1503 Found 372,1501.

[0140] Example 9 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-butyl-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-0-11).

[0141] 1) Synthesis of 1-butyl-1H-imidazol-2-carboxaldehyde (BI-07):

[0142]

[0143] The compound BI-02 was prepared according to the synthetic method described above. Using imidazole-2-carboxaldehyde (400 mg, 4.16 mmol), DMF (10 mL), potassium carbonate (690.4 mg, 5.00 mmol), and 1-iodobutane (849.2 mg, 5.00 mmol) as starting materials, and DMF (10 mL), the reaction was carried out in an oil bath at 50 °C with stirring for 6 h. 362.4 mg of an oily liquid was obtained, with a yield of 63%. 1 H NMR(500MHz, CDCl3)δ9.81(s,1H),7.26(s,1H),7.13(s,1H),4.44–4.30(m,2H),1.72(ddd,J=1 5.0,11.2,7.5Hz,2H),1.30(dd,J=15.2,7.5Hz,2H),0.90(t,J=7.4Hz,3H).MS(ESI)m / z:[M+H] + Calcd for C8H 13 N2O: 153.1022, Found: 152.73.

[0144] 2) Synthesis of the target product:

[0145]

[0146] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), 1-butyl-1H-imidazolium-2-carboxaldehyde (140 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 50 mL of water, let stand for 2 h, filter, and dry. Sonicate the mixture with ethyl acetate, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 97 mg of a yellow solid, yield 46.97%. Melting point: 163-165 °C. 1 H NMR (500MHz, DMSO-d6) δ12.33(s,1H),10.40(s,1H),7.61–7.55(m,2H),7.49–7.40(m,3H),7.40–7.33(m,1H),7.27(d,J=1.3Hz,1 H),6.86(s,1H),6.61(d,J=1.1Hz,1H),4.17(t,J=7.1Hz,2H),1.74–1.63(m,2H),1.34–1.21(m,2H),0.91(td,J=7.3,1.1Hz,3H).13 C NMR(125MHz,DMSO-d6)δ157.33,157.22,143.38,133.43,129.94,129.20,129.16,128.91 ,128.76,126.58,122.44,115.88,96.08,45.52,33.24,19.66,13.92.MS(ESI)m / z:[M+H] + Calcd for C 19 H 21 N4O2: 337.1659, Found 337.1656.

[0147] Example 10 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-benzyl-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-0-13).

[0148]

[0149] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), pyridinaldehyde (98.7 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place under nitrogen protection, and in an oil bath at 70 °C. Stir and react for 24 h. After the reaction is complete, drop the reaction solution into water (50 mL), let stand for 2 h, filter, and dry. Slurry the mixture with ethyl acetate, place in a refrigerator at 4 °C, let stand for 2 h, filter, and obtain 77 mg of yellow solid, yield 35.16%. Melting point: 260-261 °C. 1 HNMR(500MHz,DMSO-d6)δ12.59(s,1H),10.42(s,1H),8.73(s,1H),7.91(s,1H),7.69(d,J=7.8 Hz,1H),7.57(d,J=7.7Hz,2H),7.47–7.31(m,4H),6.85(s,1H),6.73(s,1H).MS(ESI)m / z:[M+H] + Calcd forC 17 H 14 N3O2: 292.1081 Found 292.1078.

[0150] Example 11 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-cyclopentane-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-1-31).

[0151] 1) Synthesis of 1-cyclopentane-1H-imidazol-2-carboxaldehyde (BI-08):

[0152]

[0153] The BI-02 was prepared according to the synthetic method described above. A 100 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (1 g, 10.4 mmol), potassium carbonate (1.73 g, 12.49 mmol), DMF (15 mL), and iodocyclopentane (2.448 g, 12.49 mmol). The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM:MeOH = 100:1) to obtain 798 mg of an oily liquid, with a yield of 46.9%. 1 H NMR (400MHz, CDCl3) δ9.80(d,J=1.5Hz,1H),7.28–7.26(m,1H),5.52(p,J=7.0Hz,1H),2.22(tdd,J=8.7,5.3,2.6Hz,2H),1.91–1.67(m,7H).

[0154] 2) Synthesis of the target product:

[0155]

[0156] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), 1-cyclopentane-1H-imidazolium-2-carboxaldehyde (151.3 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, drop the reaction mixture into 50 mL of water, let it stand for 2 h, filter, and dry. Sonicate the mixture with ethyl acetate, let it stand in a 4 °C refrigerator for 2 h, filter, and obtain 91 mg of a yellow solid, yield 52.68%. Melting point: 195-196 °C. 1H NMR (500MHz, DMSO-d6) δ12.35(d,J=2.7Hz,1H),10.38(s,1H),7.55(dd,J=8.1,2.3Hz,2H),7.50(q,J=1.4Hz,1H),7.46–7.40(m,2H),7.37–7.31 (m,1H),7.27(q,J=1.4Hz,1H),6.83(d,J=2.6Hz,1H),6.68(d,J=2.6Hz,1H),4.96–4.78(m,1H),2.50(dt,J=3.5,1.8Hz,1H),1.85–1.66(m,7H). 13 C NMR(125MHz,DMSO-d6)δ157.33,157.27,143.57,133.41,129.94,129.26(d,J=2.1H z),129.16,128.76,119.04,115.86,96.31,56.55,33.68,23.97.MS(ESI)m / z:[M+H] + Calcd for C 20 H 21 N4O2: 349.1659, Found 349.1657.

[0157] Example 12 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-sec-butyl-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-1-33).

[0158] 1) Synthesis of 1-sec-butyl-1H-imidazol-2-carboxaldehyde (BI-09):

[0159]

[0160] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (1.5 g, 15.6 mmol), potassium carbonate (2.59 g, 18.73 mmol), DMF (18 mL), and 2-iodobutane (3.44 g, 18.73 mmol). The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to obtain 617.5 mg of solid, with a yield of 26%. 1H NMR(400MHz, CDCl3) δ9.84(d,J=1.0Hz,1H),7.32(s,1H),7.29(d,J=1.1Hz,1H) ,5.33(dt,J=13.9,7.0Hz,1H),1.84–1.71(m,3H),0.83(td,J=7.4,0.9Hz,5H).

[0161] 2) Synthesis of the target product:

[0162]

[0163] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (200 mg, 0.82 mmol), 1-sec-propyl-1H-imidazolium-2-carboxaldehyde (187 mg, 1.23 mmol), cesium carbonate (400.2 mg, 1.22 mmol), anhydrous sodium sulfate (233 mg, 1.64 mmol), and DMF (10 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 50 mL of water, let stand for 2 h, filter, and dry. Slurry the mixture with ethyl acetate, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 190 mg of a yellow solid, yield 69.09%. Melting point: 176-177 °C. 1 H NMR (500MHz, DMSO-d6) δ12.39(s,1H),10.37(s,1H),7.57–7.51(m,3H),7.45–7.39(m,2H),7.36–7.28(m,2H),6.83(d,J=2.5Hz,1 H), 6.64 (d, J = 3.0Hz, 1H), 4.50 (d, J = 6.9Hz, 1H), 1.76 (td, J = 7.4, 4.2Hz, 2H), 1.39 (d, J = 6.4Hz, 3H), 0.72 (td, J = 7.7, 2.4Hz, 3H). 13 C NMR(125MHz,DMSO-d6)δ157.32(d,J=5.7Hz),129.93,129.48,129.35,129.16, 128.75,126.60,115.85,96.15,53.13,30.24,21.84,10.81.MS(ESI)m / z:[M+H] + Calcd forC 19 H 21 N4O2: 337.1659, Found 337.1658.

[0164] Example 13 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-(trimethylsilyl)-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-1-35).

[0165] 1) Synthesis of 1-(trimethylsilyl)-1H-imidazol-2-carboxaldehyde (BI-10):

[0166]

[0167] The product was prepared according to the synthesis method of BI-02. A 25 mL dry round-bottom flask was used, and imidazole-2-carboxaldehyde (1.0 g, 10.4 mmol), potassium carbonate (1.73 g, 12.6 mmol), DMF (15 mL), and 1-iodo-2-trimethylsilyl (2.7 g, 12.6 mmol) in DMF (15 mL) were added sequentially. The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to give 1.358 g of solid, with a yield of 71.58%. 1 HNMR(500MHz,DCCl3)δ9.79(d,J=0.9Hz,1H),7.27(d,J=1.0Hz,1H),7.02(s,1H),4.15(s,3H),0.04(s,9H).MS(ESI)m / z:[M+H] + Calcd for C8H 15 N2OSi:183.0948, Found:182.10.

[0168] 2) Synthesis of the target product:

[0169]

[0170] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (120 mg, 0.49 mmol), 1-(trimethylsilyl)-1H-imidazolium-2-carboxaldehyde (134.34 mg, 0.74 mmol), cesium carbonate (240 mg, 0.74 mmol), anhydrous sodium sulfate (139.57 mg, 0.98 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, evaporate to dryness, add ethyl acetate, and freeze in petroleum ether for 18 days. Filter and dry. 1HNMR(500MHz,DMSO-d6)δ12.30(s,1H),7.56(d,J=7.7Hz,2H),7.41(t,J=7.6Hz,2H),7.34–7.30(m,1H),7 .25(d,J=1.2Hz,1H),7.23(d,J=1.1Hz,1H),6.81(s,1H),6.60(s,1H),3.83(s,2H),0.03(d,J=1.0Hz,9H). 13 CNMR(125MHz,DMSO-d6)δ157.35,157.30,143.47,133.57,129.91,129.13,128.83 ,128.65,128.59,126.88,122.80,115.63,96.67,37.59,-2.18.MS(ESI)m / z:[M+H] + Calcd for C 19 H 23 N4O2Si:367.1585,Found 367.1589.

[0171] Example 14 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-isobutyl-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-1-36).

[0172] 1) Synthesis of 1-isobutyl-1H-imidazol-2-carboxaldehyde (BI-11):

[0173]

[0174] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (1.5 g, 15.6 mmol), potassium carbonate (2.59 g, 18.73 mmol), DMF (18 mL), and 2-methyliodopropane (3.44 g, 18.73 mmol). The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to give 1.06 g of solid, with a yield of 44.5%. 1 H NMR(400MHz, CDCl3)δ9.31(q,J=1.2Hz,1H),6.78(q,J=1.6Hz,1H),6.63–6.62(m,1H),3.7 1(dd,J=7.4,1.8Hz,2H), 1.57(ddd,J=11.6,7.9,5.7Hz,1H), 0.41(dd,J=7.0,2.0Hz,6H).

[0175] 2) Synthesis of the target product:

[0176]

[0177] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), 1-isobutyl-1H-imidazolium-2-carboxaldehyde (140 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 50 mL of water, let stand for 2 h, filter, and dry. Slurry the mixture with ethyl acetate, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 68 mg of yellow solid, yield 32.88%. Melting point: 203-205 °C. 1 H NMR (500MHz, DMSO-d6) δ12.33(d,J=11.5Hz,1H),10.36(d,J=11.2Hz,1H),7.55(t,J=6.2Hz,2H),7.47–7.28(m,4H),7.28 –7.22(m,1H),6.85–6.78(m,1H),6.61–6.55(m,1H),3.96(q,J=7.1Hz,2H),1.97(t,J=6.7Hz,1H),0.84(q,J=6.6Hz,6H). 13 C NMR(125MHz,DMSO-d6)δ157.33,157.23,143.70,133.42,129.93,129.19,129.15,128 .80,128.76,126.55,122.93,115.88,96.26,52.68,30.30,19.96.MS(ESI)m / z:[M+H] + Calcd for C 19 H 21 N4O2: 337.1659, Found 337.1661.

[0178] Example 15 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-(2-methoxyethyl)-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-1-38).

[0179] 1) Synthesis of 1-(2-methoxyethyl)-1H-imidazol-2-carboxaldehyde (BI-12):

[0180]

[0181] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was used, and imidazole-2-carboxaldehyde (1.5 g, 15.6 mmol), potassium carbonate (2.59 g, 18.73 mmol), DMF (18 mL), and 1-iodo-2-methoxyethane (3.48 g, 18.73 mmol) in DMF were added sequentially. The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to obtain 1.15 g of solid, with a yield of 47.0%. 1 H NMR (400MHz, CDCl3) δ9.33(dq,J=2.1,0.9Hz,1H),6.81–6.77(m,2H),4.11(td,J=5.1,1.8Hz,2H),3.19(td,J=5.0,1.7Hz,2H),2.85–2.82(m,3H).

[0182] 2) Synthesis of the target product:

[0183]

[0184] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), 1-(2-methoxyethyl)-1H-imidazolium-2-carboxaldehyde (142 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (4 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 40 mL of water, let stand for 2 h, filter, and dry. Sonicate the mixture with ethyl acetate, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 93 mg of a yellow solid, yield 44.75%. Melting point: 183-184 °C. 1 H NMR(500MHz, DMSO-d6)δ12.29(d,J=10.4Hz,1H),10.35(d,J=9.4Hz,1H),7.58–7.50(m,2H),7.46–7.29(m,4H),7.26–7 .21(m,1H),6.82(d,J=10.8Hz,1H),6.66–6.60(m,1H),4.31(p,J=5.0Hz,2H),3.59(p,J=5.0Hz,2H),3.25–3.16(m,3H). 13C NMR(125MHz,DMSO-d6)δ157.32,157.24,143.87,133.43,129.93,129.82,129.16,129.07 ,128.82,128.75,126.58,122.68,115.81,96.60,71.72,58.65,45.78.MS(ESI)m / z:[M+H] + Calcd for C 18 H 19 N4O3:339.1452,Found 339.1455.

[0185] Example 16 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-(2-methylallyl)-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-1-39).

[0186] 1) Synthesis of 1-(2-methylallyl)-1H-imidazol-2-carboxaldehyde (BI-13):

[0187]

[0188] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (1 g, 10.4 mmol), potassium carbonate (1.73 g, 12.5 mmol), DMF (15 mL), and 3-chloro-2-methyl-1-propene (1.13 g, 12.5 mmol) in DMF. The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to give 0.91 g of solid, with a yield of 58.3%. 1 HNMR(400MHz, CDCl3)δ9.40–9.33(m,1H),6.91–6.81(m,1H),6.74–6.67(m,1H), 4.57–4.51(m,2H),4.48(d,J=9.5Hz,1H),4.18–4.13(m,1H),1.31–1.25(m,3H).

[0189] 2) Synthesis of the target product:

[0190]

[0191] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), 1-(2-methylallyl)-1H-imidazolium-2-carboxaldehyde (140.2 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 50 mL of water, let stand for 2 h, filter, and dry. Slurry the mixture with ethyl acetate, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 115 mg of yellow solid, yield 56.10%. Melting point: 216-218 °C. 1 HNMR(500MHz,DMSO-d6)δ12.27(d,J=7.3Hz,1H),10.36(d,J=6.5Hz,1H),7.58–7.50(m,2H),7.46–7.30(m,4H),7.28(d,J=7.3Hz,1H ),6.85–6.79(m,1H),6.52(d,J=8.3Hz,1H),4.93–4.85(m,1H),4.77(d,J=6.5Hz,2H),4.45(d,J=6.3Hz,1H),1.67(d,J=6.9Hz,3H). 13 CNMR(125MHz,DMSO-d6)δ157.35,157.14,143.98,141.99,133.42,129.93,129.31,129.15 ,128.90,128.76,126.55,123.06,115.90,112.04,96.22,51.24,20.12.MS(ESI)m / z:[M+H] + Calcd for C 19 H 19 N4O2: 335.1503, Found 335.1505.

[0192] Example 17 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-but-2-en-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-1-42).

[0193] 1) Synthesis of (E)-1-(but-2-ene)-1H-imidazol-2-carboxaldehyde (BI-14):

[0194]

[0195] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (1 g, 10.4 mmol), potassium carbonate (1.73 g, 12.5 mmol), DMF (15 mL), and 3-chloro-2-methyl-1-propene (1.13 g, 12.5 mmol) in DMF. The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to give 0.91 g of solid, with a yield of 58.3%. 1 HNMR (400MHz, CDCl3) δ9.36–9.31(m,1H),6.85–6.67(m,2H),5.29–5.05(m,2H),4.47(dt,J=8.7,3.5Hz,2H),1.33–1.21(m,3H).

[0196] 2) Synthesis of the target product:

[0197]

[0198] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), (E)-1-(but-2-ene)-1H-imidazolium-2-carboxaldehyde (138 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 50 mL of water, let stand for 2 h, filter, and dry. Slurry the mixture with ethyl acetate, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 113 mg of yellow solid, yield 55.12%. Melting point: 189-191 °C. 1 HNMR(500MHz,DMSO-d6)δ12.26(s,1H),10.37(s,1H),7.55(d,J=7.6Hz,2H),7.46–7.31(m,4H),7.25(d,J=1.3Hz ,1H),6.83(s,1H),6.57(d,J=1.1Hz,1H),5.58(d,J=15.9Hz,2H),4.78–4.71(m,2H),1.65(dt,J=6.1,1.3Hz,3H). 13C NMR(125MHz,DMSO-d6)δ157.34,157.18,143.46,133.43,129.93,129.21,129.16,129 .01,128.95,128.76,127.07,122.32,115.88,96.29,47.45,17.81.MS(ESI)m / z:[M+H] + Calcd for C 19 H 19 N4O2: 335.1503, Found 335.1505.

[0199] Example 18 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-(3-methylbut-2-ene)-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-1-43).

[0200] 1) Synthesis of 1-(3-methylbut-2-ene)-1H-imidazol-2-carboxaldehyde (BI-15):

[0201]

[0202] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (1 g, 10.4 mmol), potassium carbonate (1.73 g, 12.5 mmol), DMF (15 mL), and 1-bromo-3-methyl-2-butene (1.86 g, 12.5 mmol) in DMF. The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to obtain 990 g of solid, with a yield of 52.3%. 1 HNMR (400MHz, CDCl3) δ9.35 (d, J=12.5Hz, 1H), 6.84–6.67 (m, 2H), 4.88 (ddd, J=9.5, 4.3, 2.9Hz, 1H), 4.58–4.51 (m, 2H), 1.35–1.28 (m, 6H).

[0203] 2) Synthesis of the target product:

[0204]

[0205] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), 1-(3-methylbut-2-ene)-1H-imidazolium-2-carboxaldehyde (151.3 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 50 mL of water, let stand for 2 h, filter, and dry. Slurry the mixture with ethyl acetate, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 129 mg of yellow solid, yield 60.28%. Melting point: 184-186 °C. 1 HNMR (500MHz, DMSO-d6) δ12.22(d,J=4.2Hz,1H),10.35(d,J=3.8Hz,1H),7.55(dt,J=9.3,2.7Hz,2H),7.47–7.31(m,4H),7.23(t,J=2.4Hz ,1H),6.83(d,J=3.9Hz,1H),6.58(d,J=4.1Hz,1H),5.33–5.20(m,1H),4.76(t,J=5.4Hz,2H),1.81(t,J=2.5Hz,3H),1.72(t,J=2.5Hz,3H). 13 C NMR(125MHz,DMSO-d6)δ157.32,157.17,143.31,137.13,133.43,129.92,129.17,129.15,129.02 ,128.91,128.77,126.57,122.17,120.19,115.86,96.44,44.02,25.77,18.34.MS(ESI)m / z:[M+H] + Calcd for C 20 H 21 N4O2: 349.1659, Found 349.1662.

[0206] Example 19 Synthesis of (3Z,6Z)-3-(benzylidene)-6-((1-(cyclopropylmethyl)-1H-imidazol-2-yl)methylene)piperazine-2,5-dione (BEPM-1-45).

[0207] 1) Synthesis of 1-(cyclopropylmethyl)-1H-imidazol-2-carboxaldehyde (BI-16):

[0208]

[0209] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (264 mg, 2.75 mmol), potassium carbonate (446 mg, 3.3 mmol), DMF (6 mL), and 1-(cyclopropylmethyl)-1H-imidazole-2-carboxaldehyde (600 mg, 3.3 mmol) in DMF. The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to obtain 274.3 mg of solid, with a yield of 66.4%. 1 HNMR(400MHz, CDCl3)δ9.38–9.27(m,1H),6.86–6.77(m,2H),3.83–3.74(m,2H),0 .78(ddtt,J=9.9,7.4,5.1,2.6Hz,1H),0.19–0.09(m,2H),-0.10(t,J=5.1Hz,2H).

[0210] 2) Synthesis of the target product:

[0211]

[0212] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-benzylpiperazine-2,5-dione (150 mg, 0.61 mmol), 1-(cyclopropylmethyl)-1H-imidazolium-2-carboxaldehyde (138 mg, 0.92 mmol), cesium carbonate (300.1 mg, 0.92 mmol), anhydrous sodium sulfate (174.5 mg, 1.22 mmol), and DMF (5 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 24 h. After the reaction is complete, add the reaction solution dropwise to 50 mL of water, let stand for 2 h, filter, and dry. Slurry the mixture with ethyl acetate, let stand in a 4 °C refrigerator for 2 h, filter, and obtain 108 mg of yellow solid, yield 52.68%. Melting point: 233-234 °C. 1 HNMR(500MHz,DMSO-d6)δ12.30(s,1H),10.38(s,1H),7.56(d,J=7.6Hz,2H),7.48(d,J=1.4Hz,1H),7.45–7.38(m,2H),7.37–7.30(m,1H),7.26(d,J =1.2Hz,1H),6.83(s,1H),6.62(d,J=2.1Hz,1H),4.03(d,J=7.0Hz,2H),1. 30–1.11(m,1H),0.53(dd,J=8.0,2.1Hz,2H),0.39(dd,J=4.8,1.8Hz,2H). 13C NMR(125MHz,DMSO-d6)δ157.40,157.29,143.32,133.52,129.94,129.23,129.14,12 8.86,128.70,126.82,122.24,115.79,96.19,49.82,12.48,4.16.MS(ESI)m / z:[M+H] + Calcd for C 19 H 19 N4O2: 335.1503, Found 335.1500.

[0213] Example 20 Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-tert-butyl-1H-imidazol)-2-methylene)piperazine-2,5-dione (EPM-1-33).

[0214] 1) Synthesis of 4-fluorophenol benzaldehyde (BI-17):

[0215]

[0216] Take a 50mL dry round-bottom flask and add 1g Molecular sieves were used, and oxygen-filled bulbs were filled with p-fluorophenol (500 mg, 4.46 mol), 3-formylphenylboronic acid (1.0 g, 6.69 mol), DCM (10 mL), copper acetate (811 mg, 4.46 mol), and TEA (135.6 mg, 1.34 mol). The mixture was stirred at room temperature for 24 h, and then subjected to TLC (PE:EA = 1:1). After the reaction was complete, the mixture was filtered, concentrated under reduced pressure, and subjected to column chromatography (EA:PE = 4:1). The concentrate was then concentrated under reduced pressure to give 220.0 mg of white solid, with a yield of 22%.

[0217] 2) Synthesis of Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (BI-18):

[0218]

[0219] Weigh 156.8 mg (0.73 mmol) of 3-p-fluorophenoxybenzaldehyde, 1,4-diacetylpiperazine-2,5-dione (150 mg, 0.60 mmol) of 1,4-diacetylpiperazine-2,5-dione (295.3 mg, 0.91 mmol) of cesium carbonate, 171.7 mg (1.21 mmol) of anhydrous sodium sulfate, and 3 mL of DMF into a dry 25 mL single-necked flask. Under nitrogen protection, the mixture was reacted at room temperature for 20 h. Thin-layer chromatography was used to determine the completion of the reaction. The reaction solution was poured into cold water, and a large amount of solid precipitated. After filtration and drying, 99.8 mg of a brown solid was obtained, with a yield of 47%, which was directly used in the next reaction.

[0220] 3) Synthesis of 1-sec-butyl-1H-imidazol-2-carboxaldehyde (BI-19):

[0221]

[0222] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (1.5 g, 15.6 mmol), potassium carbonate (2.59 g, 18.73 mmol), DMF (18 mL), and 2-iodobutane (3.44 g, 18.73 mmol). The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to obtain 617.5 mg of solid, with a yield of 26%. 1 H NMR(400MHz, CDCl3) δ9.84(d,J=1.0Hz,1H),7.32(s,1H),7.29(d,J=1.1Hz,1H) ,5.33(dt,J=13.9,7.0Hz,1H),1.84–1.71(m,3H),0.83(td,J=7.4,0.9Hz,5H).

[0223] 4)(3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-sec-butyl-1H-imidazol)-2-methylene)piperazine-2,5-dione

[0224]

[0225] The compound BEPM-0-1 was prepared according to the synthetic method described above. Starting materials included 150 mg (0.34 mmol) of 1-(3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione, 96.6 mg (0.64 mmol) of 1-sec-butyl-1H-imidazolium-2-carboxaldehyde, 207 mg (0.64 mmol) of cesium carbonate, and 120 mg (0.85 mmol) of anhydrous sodium sulfate. The mixture was reacted in an oil bath at 50 °C for 24 h under nitrogen protection in DMF (5 mL). The mixture was ultrasonically beaten with ethyl acetate to give 80.1 mg of a yellow solid, yielding 42.3%. Melting point: 226-228 °C. 1H NMR (400MHz, DMSO-d6))) δ12.29(s,1H),7.66–7.35(m,4H),7.29(ddd,J=13.0,7.5,5.2Hz,3H),7.16(ddd,J=10.6,5.4,2.8Hz,2H),6.90(d,J=8 .1Hz,1H),6.70(s,1H),6.58(s,1H),4.52(t,J=7.2Hz,1H),1.81(d,J=8.2Hz,2H),1.48–1.41(m,3H),0.78(q,J=7.6Hz,3H).MS(ESI)m / z:[M+H] + Calcd for C 25 H 24 FN4O3:447.1827,Found:447.1825.

[0226] Example 21 Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-isobutyl-1H-imidazol)-2-methylene)piperazine-2,5-dione (EPM-1-36).

[0227] 1) Synthesis of 1-isobutyl-1H-imidazol-2-carboxaldehyde (BI-20):

[0228]

[0229] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (1.5 g, 15.6 mmol), potassium carbonate (2.59 g, 18.73 mmol), DMF (18 mL), and 2-methyliodopropane (3.44 g, 18.73 mmol). The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to give 1.06 g of solid, with a yield of 44.5%. 1 H NMR(400MHz, CDCl3)δ9.31(q,J=1.2Hz,1H),6.78(q,J=1.6Hz,1H),6.63–6.62(m,1H),3.7 1(dd,J=7.4,1.8Hz,2H), 1.57(ddd,J=11.6,7.9,5.7Hz,1H), 0.41(dd,J=7.0,2.0Hz,6H).

[0230] 2. Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-isobutyl-1H-imidazol)-2-methylene)piperazine-2,5-dione:

[0231]

[0232] The compound BEPM-0-1 was prepared according to the synthetic method described above. Starting materials included Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (150 mg, 15.6 mmol), 1-isobutyl-1H-imidazolium-formaldehyde (96.6 mg, 0.64 mmol), cesium carbonate (207 mg, 0.64 mmol), and anhydrous sodium sulfate (120 mg, 0.85 mmol). The mixture was reacted in DMF (4 mL) under nitrogen purging, stirred in an oil bath at 50 °C for 24 h. The mixture was ultrasonically beaten with ethyl acetate to give 93 mg of a yellow solid, yielding 59.86%. Melting point: 178-180 °C. 1 H NMR (400MHz, DMSO-d6)) δ12.35(s,1H),10.43(s,1H),7.39(q,J=6.1Hz,2H),7.31–7.16(m,5H),7.16–7.06(m,2H),6.9 4–6.87(m,1H),6.81–6.75(m,1H),6.61–6.52(m,1H),3.96(q,J=7.2Hz,2H),2.04–1.89(m,1H),0.83(q,J=6.5Hz,6H). 13 CNMR(125MHz,DMSO-d6)δ159.61,157.71,157.49,157.29,157.20,153.04,143.70,135.46,130.74,129.19,128.80,127.40 ,125.10,122.93,121.05,120.98,119.52,118.43,117.11,116.93,114.98,96.34,52.67,30.29,19.95.MS(ESI)m / z:[M+H] + Calcd for C 25 H 24 FN4O3:447.1827,Found:447.1830.

[0233] Example 22 Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-(2-methoxyethyl)-1H-imidazolium)-2-methylene)piperazine-2,5-dione (EPM-1-38).

[0234] 1) Synthesis of 1-(2-methoxyethyl)-1H-imidazol-2-carboxaldehyde (BI-21):

[0235]

[0236] The product was prepared according to the synthesis method of BI-02. A 25 mL dry round-bottom flask was used, and imidazole-2-carboxaldehyde (1.5 g, 15.6 mmol), potassium carbonate (2.59 g, 18.73 mmol), DMF (18 mL), and 1-iodo-2-methoxyethane (3.48 g, 18.73 mmol) in DMF were added sequentially. The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to give 1.15 g of solid, with a yield of 47.0%. 1 H NMR (400MHz, CDCl3) δ9.33(dq,J=2.1,0.9Hz,1H),6.81–6.77(m,2H),4.11(td,J=5.1,1.8Hz,2H),3.19(td,J=5.0,1.7Hz,2H),2.85–2.82(m,3H).

[0237] 2. Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-(2-methoxyethyl))-1H-imidazolium)-2-methylene)piperazine-2,5-dione:

[0238]

[0239] The compound BEPM-0-1 was prepared according to the synthetic method described above. Starting materials included Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (120 mg, 0.34 mmol), 2-methoxyethyl)imidazolium-2-carboxaldehyde (78.32 mg, 0.5 mmol), cesium carbonate (165.5 mg, 0.5 mmol), and anhydrous sodium sulfate (96.2 mg, 0.68 mmol). The mixture was reacted in DMF (4 mL) under nitrogen purging, stirred in an oil bath at 70 °C for 24 h. The mixture was then ultrasonically beaten with ethyl acetate to give 43 mg of a yellow solid, with a yield of 27.8%. Melting point: 155-157 °C. 1 H NMR (400MHz, DMSO-d6) δ12.29(d,J=4.4Hz,1H),10.42(s,1H),7.45–7.36(m,2H),7.31–7.17(m,5H),7.12(ddd,J=8.7,4.5,1.9Hz, 2H),6.94–6.88(m,1H),6.81–6.77(m,1H),6.63(d,J=2.9Hz,1H),4.32(t,J=5.2Hz,2H),3.60(t,J=5.1Hz,2H),3.23–3.19(m,3H).13 C NMR (125MHz, DMSO-d6) δ159.62,157.71,157.50,157.30,157.19,153.06,153.04,143.86,135.47,130.75,129.07,128.82,127 .41,125.09,122.68,121.06,120.99,119.52,118.43,117.11,116.93,114.93,96.68,71.72,58.64,45.78.MS(ESI)m / z:[M+H] + Calcd for C 24 H 22 FN4O4:449.1620,Found:449.1621.

[0240] Example 23 Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-(2-methylallyl)-1H-imidazol)-2-methylene)piperazine-2,5-dione (EPM-1-39).

[0241] 1) Synthesis of 1-(2-methylallyl)-1H-imidazol-2-carboxaldehyde (BI-22):

[0242]

[0243] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (1 g, 10.4 mmol), potassium carbonate (1.73 g, 12.5 mmol), DMF (15 mL), and 3-chloro-2-methyl-1-propene (1.13 g, 12.5 mmol) in DMF. The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to give 0.91 g of solid, with a yield of 58.3%. 1 HNMR(400MHz, CDCl3)δ9.40–9.33(m,1H),6.91–6.81(m,1H),6.74–6.67(m,1H), 4.57–4.51(m,2H),4.48(d,J=9.5Hz,1H),4.18–4.13(m,1H),1.31–1.25(m,3H).

[0244] 2. Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-(2-methylallyl)-1H-imidazol)-2-methylene)piperazine-2,5-dione:

[0245]

[0246] The compound BEPM-0-1 was prepared according to the synthetic method described above. Starting materials included Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (120 mg, 0.34 mmol), 2-methoxyethyl)imidazolium-2-carboxaldehyde (78.32 mg, 0.5 mmol), cesium carbonate (165.5 mg, 0.5 mmol), and anhydrous sodium sulfate (96.2 mg, 0.68 mmol). The mixture was reacted in DMF (4 mL) under nitrogen purging, stirred in an oil bath at 70 °C for 24 h. The mixture was then ultrasonically beaten with ethyl acetate to give 43 mg of a yellow solid, with a yield of 27.8%. Melting point: 174-176 °C. 1 H NMR(400MHz,DMSO-d6)δ12.28(s,1H),10.43(s,1H),7.44–7.35(m,2H),7.29–7.18(m,5H),7.14–7.09(m,2 H),6.94–6.89(m,1H),6.79(s,1H),6.52(s,1H),4.92–4.86(m,1H),4.77(s,2H),4.45(s,1H),1.68(s,3H). 13 C NMR (125MHz, DMSO-d6) δ159.62,157.71,157.50,157.18,153.04,153.02,143.96,141.98,135.43,130.75,129.29,128.91,127 .31,125.09,123.07,121.06,120.99,119.51,118.44,117.11,116.93,115.04,112.05,96.33,51.24,20.11.MS(ESI)m / z:[M+H] + Calcd for C 25 H 22 FN4O3:445.1670,Found:445.1674.

[0247] Example 24 Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-(but-2-ene)-1H-imidazolium)-2-methylene)piperazine-2,5-dione (EPM-1-42).

[0248] 1) Synthesis of (E)-1-(but-2-ene)-1H-imidazol-2-carboxaldehyde (BI-23):

[0249]

[0250] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (1 g, 10.4 mmol), potassium carbonate (1.73 g, 12.5 mmol), DMF (15 mL), and 3-chloro-2-methyl-1-propene (1.13 g, 12.5 mmol) in DMF. The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to give 0.91 g of solid, with a yield of 58.3%. 1 HNMR (400MHz, CDCl3) δ9.36–9.31(m,1H),6.85–6.67(m,2H),5.29–5.05(m,2H),4.47(dt,J=8.7,3.5Hz,2H),1.33–1.21(m,3H).

[0251] 2)(3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((E)-1-(but-2-ene)-1H-imidazolium)-2-methylene)piperazine-2,5-dione.

[0252]

[0253] The compound BEPM-0-1 was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (120 mg, 0.34 mmol), (E)-1-(but-2-ene)-1H-imidazolium-2-carboxaldehyde (83.42 mg, 0.5 mmol), cesium carbonate (165.5 mg, 0.5 mmol), and anhydrous sodium sulfate (96.2 mg, 0.68 mmol). The mixture was reacted in DMF (4 mL) under nitrogen purging, stirred in an oil bath at 70 °C for 24 h. The mixture was then ultrasonically beaten with ethyl acetate to give 47.1 mg of a yellow solid, yielding 31.4%. Melting point: 172-174 °C. 1 H NMR (400MHz, DMSO-d6) δ12.26(s,1H),10.43(s,1H),7.44–7.36(m,2H),7.31–7.17(m,5H),7.15–7.10(m,2H),6.92(d d,J=8.2,2.5Hz,1H),6.79(s,1H),6.57(d,J=2.4Hz,1H),5.65–5.45(m,2H),4.74(d,J=5.3Hz,2H),1.81–1.62(m,3H). 13C NMR (125MHz, DMSO-d6) δ159.62,157.71,157.50,157.22,157.19,153.05,153.03,143.44,135.44,130.76,129.19,129.01,128.95 ,127.07,125.09,122.33,121.06,121.00,119.51,118.45,117.12,116.93,115.01,96.39,47.45,17.80,13.36.MS(ESI)m / z:[M+H] + Calcd for C 25 H 22 FN4O3:445.1670,Found:445.1672.

[0254] Example 25 Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-(3-methylbut-2-ene)-1H-imidazolium)-2-methylene)piperazine-2,5-dione (EPM-1-43).

[0255] 1) Synthesis of 1-(3-methylbut-2-ene)-1H-imidazol-2-carboxaldehyde (BI-24):

[0256]

[0257] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (1 g, 10.4 mmol), potassium carbonate (1.73 g, 12.5 mmol), DMF (15 mL), and 1-bromo-3-methyl-2-butene (1.86 g, 12.5 mmol) in DMF. The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to obtain 990 g of solid, with a yield of 52.3%. 1 HNMR (400MHz, CDCl3) δ9.35 (d, J=12.5Hz, 1H), 6.84–6.67 (m, 2H), 4.88 (ddd, J=9.5, 4.3, 2.9Hz, 1H), 4.58–4.51 (m, 2H), 1.35–1.28 (m, 6H).

[0258] 2. Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-(3-methylbut-2-ene)-1H-imidazolium)-2-methylene)piperazine-2,5-dione:

[0259]

[0260] The compound BEPM-0-1 was prepared according to the synthetic method described above. Starting materials included Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (120 mg, 0.34 mmol), 1-(3-methylbut-2-ene)-1H-imidazolium-2-carboxaldehyde (83.42 mg, 0.5 mmol), cesium carbonate (165.5 mg, 0.5 mmol), and anhydrous sodium sulfate (96.2 mg, 0.68 mmol). The mixture was reacted in an oil bath at 70 °C for 24 h under nitrogen protection in DMF (4 mL). The mixture was ultrasonically beaten with ethyl acetate to give 47.1 mg of a yellow solid, with a yield of 30.2%. Melting point: 182-184 °C. 1 H NMR (400MHz, DMSO-d6) δ12.28–12.17(m,1H),10.42(s,1H),7.41(dd,J=7.9,1.9Hz,1H),7.36(d,J=1.8Hz,1H),7.30–7.18(m,5H),7.12(ddd,J=9.0 ,4.5,2.0Hz,2H),6.94–6.89(m,1H),6.79(d,J=1.9Hz,1H),6.58(d,J=1. 9Hz, 1H), 5.24 (d, J = 1.7Hz, 1H), 4.76 (d, J = 7.1Hz, 2H), 1.83–1.66 (m, 6H). 13 CNMR(125MHz,DMSO-d6)δ159.62,157.71,157.51,157.22,157.18,153.05,153.03,143.30,137.14,135.46,130.76,129.02,128.91,12 7.39,125.07,122.17,121.06,121.00,120.18,119.52,118.44,117.12,116.93,114.98,96.52,44.02,25.76,18.33.MS(ESI)m / z:[M+H] + Calcd for C 26 H 24 FN4O3:459.1827,Found:459.1828.

[0261] Example 26 Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-(prop-2-ynyl)-1H-imidazol)-2-methylene)piperazine-2,5-dione (EPM-1-34).

[0262] 1) 1-(prop-2-ynyl)-1H-imidazol-2-carboxaldehyde (BI-25):

[0263]

[0264] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was used, and imidazole-2-carboxaldehyde (1.5 g, 15.6 mmol), potassium carbonate (2.59 g, 18.73 mmol), DMF (18 mL), and 3-bromo-1-yne (2.23 g, 18.73 mmol) in DMF were added sequentially. The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to give 1 g of solid, with a yield of 47.8%. 1 H NMR (400MHz, CDCl3) δ9.35–9.31(m,1H),6.99(q,J=1.8Hz,1H),6.83(q,J=1.4Hz,1H),4.77(t,J=2.1Hz,2H),2.05(dt,J=4.3,2.7Hz,1H).

[0265] 2) Synthesis of (Z)-1-acetyl-3-((1-(prop-2-yne)-1H-imidazolium)-2-methylene)piperazine-2,5-dione (BI-26):

[0266]

[0267] Take a 25 mL dry round-bottom flask and add 1-(prop-2-yn-1-yl)-1H-imidazol-2-carboxaldehyde (100 mg, 0.75 mmol), 1,4-diacetylpiperazine-2,5-dione (295.5 mg, 1.5 mmol), cesium carbonate (364.3 mg, 1.12 mmol), and DMF (5 mL) sequentially. React at room temperature in the dark for 5 h. Wash the organic phase with water, slurry with ethyl acetate, and dry to obtain a yellow solid, which can be used directly in the next reaction. Yield: 59.1%. 1 H NMR (400MHz, DMSO-d6) δ12.16(s,1H),10.48(s,1H),7.49–7.37(m,2H),7.25(td,J=13.9,6.3Hz,5H),7.12(dh, J=7.1,3.2Hz,2H),6.96–6.89(m,1H),6.83–6.77(m,1H),6.71–6.66(m,1H),5.16(s,2H),3.58(d,J=2.6Hz,1H).

[0268] 3)(3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-(prop-2-ynyl)-1H-imidazol)-2-methylene)piperazine-2,5-dione:

[0269]

[0270] The reaction mixture was prepared in an oil bath at 50°C for 22 h under nitrogen protection, using (Z)-1-acetyl-3-((1-(propion-2-ynyl)imidazolium)-2-methylene)piperazine-2,5-dione (107 mg, 0.39 mmol), 3-(p-fluorophenoxy)benzaldehyde (102 mg, 0.47 mmol), cesium carbonate (192 mg, 0.59 mmol), and DMF (5 mL). The mixture was ultrasonically beaten with ethyl acetate to give 74 mg of a yellow solid, yielding 45.6%. Melting point: 194-196°C. 1 H NMR (400MHz, DMSO-d6) δ12.16(s,1H),10.48(s,1H),7.49–7.37(m,2H),7.25(td,J=13.9,6.3Hz,5H),7.12(dh, J=7.1,3.2Hz,2H),6.96–6.89(m,1H),6.83–6.77(m,1H),6.71–6.66(m,1H),5.16(s,2H),3.58(d,J=2.6Hz,1H). 13 C NMR(125MHz,DMSO-d6)δ159.62,157.71,157.50,157.25,157.16,153.06,153.04,143.60,135.44,130.75,129.57,129.01 ,125.12,122.22,121.06,121.00,119.55,118.46,117.12,116.93,115.14,96.06,78.89,77.14,35.44.MS(ESI)m / z:[M+H] + Calcd for C 24 H 18 FN4O3:429.1357,Found:429.1361.

[0271] Example 27 Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-(but-2-ene)-1H-imidazolium)-2-methylene)piperazine-2,5-dione (EPM-1-45).

[0272] 1) Synthesis of 1-(cyclopropylmethyl)-1H-imidazol-2-carboxaldehyde (BI-27):

[0273]

[0274] The BI-02 was prepared according to the synthetic method described above. A 25 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (264 mg, 2.75 mmol), potassium carbonate (446 mg, 3.3 mmol), DMF (6 mL), and 1-(cyclopropylmethyl)-1H-imidazole-2-carboxaldehyde (600 mg, 3.3 mmol) in DMF. The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (PE:EA = 4:1) to obtain 274.3 mg of solid, with a yield of 66.4%. 1 HNMR(400MHz, CDCl3)δ9.38–9.27(m,1H),6.86–6.77(m,2H),3.83–3.74(m,2H),0 .78(ddtt,J=9.9,7.4,5.1,2.6Hz,1H),0.19–0.09(m,2H),-0.10(t,J=5.1Hz,2H).

[0275] 2)(3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-(cyclopropylmethyl)-1H-imidazol)-2-methylene)piperazine-2,5-dione:

[0276]

[0277] The compound BEPM-0-1 was prepared according to the synthetic method described above. Starting materials included Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (120 mg, 0.34 mmol), 1-(cyclopropylmethyl)-1H-imidazolium-2-carboxaldehyde (76.3 mg, 0.5 mmol), cesium carbonate (165.5 mg, 0.5 mmol), and anhydrous sodium sulfate (96.2 mg, 0.68 mmol). The mixture was reacted in DMF (4 mL) under nitrogen purging, stirred in an oil bath at 70 °C for 24 h. The mixture was then ultrasonically beaten with ethyl acetate to give 108 mg of a yellow solid, yield 72%. Melting point: 201-202 °C. 1H NMR (400MHz, DMSO-d6) δ12.30(s,1H),10.44(s,1H),7.48(d,J=1.4Hz,1H),7.42(t,J=8.0Hz,1H),7.31–7.18(m,5H),7.15–7.10(m,2H),6.92(dd,J= 8.2,2.5Hz,1H),6.80(d,J=2.2Hz,1H),6.62(d,J=2.2Hz,1H),4.06–4.00( m,2H),1.20–1.12(m,1H),0.57–0.50(m,2H),0.38(dt,J=5.4,2.6Hz,2H). 13 C NMR (125MHz, DMSO-d6) δ159.62,157.71,157.50,157.26,157.16,153.05,153.03,143.27,135.43,130.76,129.11,1 28.89,127.34,125.10,122.28,121.06,121.00,119.52,118.45,117.11,116.93,115.01,96.42,49.82,12.47,4.15.

[0278] Example 28 Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-cyclopentane-1H-imidazol)-2-methylene)piperazine-2,5-dione (EPM-1-31).

[0279] 1) Synthesis of 1-cyclopentane-1H-imidazol-2-carboxaldehyde (BI-28):

[0280]

[0281] The BI-02 was prepared according to the synthetic method described above. A 100 mL dry round-bottom flask was filled sequentially with imidazole-2-carboxaldehyde (1 g, 10.4 mmol), potassium carbonate (1.73 g, 12.49 mmol), DMF (15 mL), and iodocyclopentane (2.448 g, 12.49 mmol). The mixture was stirred in an oil bath at 50 °C for 6 h. The mixture was filtered, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM:MeOH = 100:1) to obtain 798 mg of an oily liquid, with a yield of 46.9%. 1HNMR (400MHz, CDCl3) δ9.80(d,J=1.5Hz,1H),7.28–7.26(m,1H),5.52(p,J=7.0Hz,1H),2.22(tdd,J=8.7,5.3,2.6Hz,2H),1.91–1.67(m,7H).

[0282] 2. Synthesis of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-cyclopentane-1H-imidazol)-2-methylene)piperazine-2,5-dione

[0283]

[0284] The compound BEPM-0-1 was prepared according to the synthetic method described above. Starting materials included 120 mg (0.34 mmol) of 1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione, 1-cyclopentane-2-carboxaldehyde (83.42 mg, 0.5 mmol), cesium carbonate (165.5 mg, 0.5 mmol), and anhydrous sodium sulfate (96.2 mg, 0.68 mmol). The mixture was reacted in an oil bath at 45 °C for 24 h under nitrogen protection in DMF (4 mL). The mixture was ultrasonically beaten with ethyl acetate to give 93 mg of a yellow solid, yielding 59.86%. Melting point: 220-222 °C. 1 H NMR(400MHz,DMSO-d6)δ12.34(s,1H),10.44(s,1H),7.50(d,J=2.0Hz,1H), 7.41(td,J=8.0,1.8Hz,1H),7.31–7.18(m,5H),7.12(ddd,J=8.9,4.5,1.8Hz ,2H),6.92(dt,J=8.1,2.2Hz,1H),6.79(d,J=1.8Hz,1H),6.67(d,J=1.7Hz,1 H), 4.92–4.82 (m, 1H), 2.12 (q, J = 6.4Hz, 2H), 1.75 (dt, J = 27.2, 10.3Hz, 6H). 13 C NMR (125MHz, DMSO-d6) δ159.61,157.71,157.49,157.32,157.18,153.05,153.03,143.56,135.44,130.75,129.26,127.38 ,125.10,121.06,120.99,119.51,119.05,118.43,117.11,116.93,114.98,96.39,56.54,33.69,23.97.MS(ESI)m / z:[M+H] +Calcd for C 26 H 24 FN4O3:459.1827,Found:459.1830.

[0285] Example 29: NCI-H460 cell proliferation inhibition experiment.

[0286] Cell viability testing steps:

[0287] 1) Seed cells into 96-well plates;

[0288] Tumor cells in the logarithmic growth phase will be digested, centrifuged, and re-vortexed with fresh culture medium before being counted using a cell counting chamber at a rate of 0.5-10 × 10⁶ cells per well. 4 To determine the standard for cells per well, add 100 μL of culture medium containing cells to each well of a 96-well plate and place the 96-well plate in an incubator (37°C, 5% CO2) for incubation.

[0289] 2) Cell-level drug administration;

[0290] After the cells have fully adhered (approximately 16-24 hours), the drug is diluted to different concentrations using serum-containing medium, with three replicates for each concentration. The medium in the 96-well plate is removed, and 100 μL of medium containing different drug concentrations is added to each well. The plate is then incubated in an incubator (37°C, 5% CO2) for 72 hours.

[0291] 3) Add MTT;

[0292] Add 20 μL of 5 mg / mL MTT to each well and incubate in an incubator for 3-4 hours. (Prepared MTT powder with PBS and filtered through a 0.22 μM sterile membrane yielded a yellow solution with a concentration of 5 mg / mL.)

[0293] 4) Detection wavelength of the ELISA reader;

[0294] Discard the culture medium and MTT. Add 200 μL of DMSO to each well. Place the 96-well plate on a 96-well plate shaker to completely dissolve the purple formazan. Then, use a microplate reader to measure the absorbance (OD value) at a wavelength of 570 nm. This experiment also requires setting up zeroing wells (blank culture medium, MTT, DMSO).

[0295] Cell inhibition rate = 1 - (OD value of drug-treated group - OD value of zero-adjustment group) / (OD value of blank group - OD value of zero-adjustment group) × 100%. Calculate the IC50 of the tested compound against NCI-H460 cell line. 50 Values. See Table 2 for specific data.

[0296] Table 2. Inhibition of NCI-H460 cell proliferation by the compounds of the present invention.

[0297]

[0298]

[0299] Note: NCI-H460 is a human lung cancer cell line, and punabulin is a positive control. The control group contains DMSO without the sample, and the blank group contains neither DMSO nor the sample.

[0300] Preferably, the effective therapeutic dose of the diketopiperazine compound is IC50. 50 The value ranges from 1 μM to 0.028 μM. It is evident that the above-mentioned compound IC of this invention... 50 All values ​​fall within this range.

[0301] Example 30: Solubility test.

[0302] 1. Salt formation steps and methods:

[0303] The compound (1.0 eq) was placed in a dry round-bottom flask, dissolved in an appropriate amount of methanol, and a hydrochloric acid-methanol solution (10.0 eq) was added dropwise. The mixture was stirred at room temperature in the dark for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure until no solvent remained. A small amount of methanol was dissolved in the solution and added dropwise to ethyl acetate. A solid precipitated out. The mixture was placed in a 0°C cold trap and stirred for 2 h. The mixture was then filtered, and the filter cake was washed with cold ethyl acetate. The filter cake was dried in a 50°C vacuum drying oven to obtain the salt-forming product.

[0304] 2. Solubility test.

[0305] 1) Preparation of the standard curve:

[0306] Weigh appropriate amounts of each compound and dissolve them in methanol to prepare a 1 mg / mL stock solution. Dilute the stock solution with methanol and water (60:40) to different concentrations of working solution, which are then used as standard samples for testing. The concentrations of the different working solutions were successively diluted as follows: 100,000, 10,000, 5,000, 2,500, 500, 250, 100, 50, 20, 10, 5, and 2 ng / mL.

[0307] 2) Method for determining the solubility of salt-forming samples:

[0308] Take a 1.5 mL brown EP tube and add 1 mL of ultrapure water. Then, add 1-2 mg of the compound to each brown EP tube, vortex, and sonicate until the compound no longer dissolves (the solution becomes turbid or contains suspended particles). Place in an incubator and maintain the temperature at (37±1)℃, at 100 rpm, and shake for 24 h to allow complete dissolution equilibrium. After 24 h, quickly filter the supernatant through a 0.22 μm microporous membrane, discard the initial filtrate, and take 500 μL of the subsequent filtrate, diluting it with 100 μL of methanol. Repeat the determination at least three times.

[0309] Inject the sample according to the chromatographic conditions, determine the peak area, and calculate the equilibrium solubility of each compound in pure water. See Table 3 for details.

[0310] Table 3. Equilibrium solubility of each compound in pure water

[0311]

[0312]

[0313] Preferably, the solubility range of the diketopiperazine hydrochloride compound is 1.14 mg / ml to 354.47 mg / ml. A solubility greater than 1 mg / ml is considered preferred, and it is evident that the solubility values ​​of the compounds described above in this invention all fall within this range.

[0314] In the above embodiments, the descriptions of each embodiment have different focuses. For parts that are not described in detail or recorded in a certain embodiment, please refer to the relevant descriptions of other embodiments.

[0315] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any modifications, equivalent substitutions and improvements made by those skilled in the art within the scope of the technology disclosed in the present invention and within the spirit and principles of the present invention should be covered within the scope of protection of the present invention.

Claims

1. A diketopiperazine compound, characterized by, The molecular formula structure general formula (I) of the diketopiperazine compound is: wherein each R1is independently selected from a hydrogen atom, an alkyl group, an alkenyl group, an alkynyl group, an aryl group, an alkoxy group, an amido group, or an acyloxy group, or R2is selected from any one of a hydrogen atom, an alkyl group, an aryl group, an aromatic heterocycle, a saturated polyheterocycle, and an unsaturated polyheterocycle.

2. The diketopiperazine compound according to claim 1, wherein The X is -O- or -C=O-.

3. The diketopiperazine compound of claim 1, wherein The R1' in the para position to the X group is a fluorine atom or a hydrogen atom.

4. The diketopiperazine compound of claim 1, wherein The R2 is one of methyl, ethyl, isopropyl and tert-butyl.

5. The diketopiperazine compound of claim 1, wherein R2is selected from one of the following groups:

6. A pharmaceutical composition, characterized by, The pharmaceutical composition comprises a therapeutically effective dose of IC 50 a diketopiperazine compound of any one of claims 1-5 having a value of 1 uM - 0.028 uM and a pharmaceutically acceptable carrier.

7. Use of the diketopiperazine compound according to any one of claims 1-5 in the preparation of a medicament for preventing and / or treating cancer.

8. Use of the pharmaceutical composition according to claim 6 in the preparation of a medicament for preventing and / or treating cancer.

9. A method of synthesizing a diketopiperazine compound, characterized by, The method for synthesizing the diketopiperazine compound as claimed in any one of claims 1-5 comprises: S1, synthesizing or providing imidazole aldehyde C; S2, synthesizing or providing intermediate 2,5-diketopiperazine derivative B; S3, synthesizing or providing 3-substituted benzaldehyde A; S4, performing Aldol condensation of 3-substituted benzaldehyde A and intermediate 2,5-diketopiperazine derivative B to obtain a compound having the structure shown in formula (I), i.e. the diketopiperazine compound.

10. The synthesis method of the diketopiperazine compound according to claim 9, wherein, in step S1, K2CO3, DMF, 50℃, 6h, 26.0%-71.58%; in step S2, Cs2CO3, Na2SO4, DMF, 45-70℃, 22h-24h, N2 protection, 25.8%-72.0%; in step S3, Cu(OAc)2, TEA, Dry DCM, O2, rt, 22.0%; in step S3, Cs2CO3, Na2SO4, DMF, rt-50℃, 6h-22h, N2 protection, 45.6%-47.0%.

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